Evaluation of lignin-based black liquor decolorization by Trametes versicolor U 80

NASA Astrophysics Data System (ADS)

Amriani, Feni; Sari, Ajeng Arum; R. Irni Fitria, A.; Abimanyu, Haznan; Tachibana, Sanro

2017-01-01

Bioethanol second generation (G-2) production process generated black liquor that need to treat before the disposal to prevent environmental pollution. Usually, coagulation technology using polyaluminium chloride was employed to precipitate dissolved lignin and intended to decolorize black liquor. However, this single work is not effective to treat black liquor, so that it requires another work to treat remain brownish liquor. Isolated fungal strain from Japan Trametes versicolor U 80 and Phanerochaete chrysosporium are white rot fungi that are known in ligninolytic enzymes secretion to biodegrade soluble lignin. Decolorization of black and brownish liquor is an indicator of fungi works since lignin is known as the colour agent in liquor colouration. This work evaluated black and brownish liquor decolorization using both fungi that correspond to fungal growth. Liquor toxicity was observed based on mycelial dry weight after 30 days incubation as the presumption of the connection of fungal growth and decolorization. The biosorption from the dead cell was also evaluated for fungal adsorption capability in black and brownish decolorization. As the result, T. versicolor U 80 was able to decolorize brownish liquor 51.5% after 21 days incubation and 68.6% black liquor at 15 days incubation. MnP and Laccase enzymes activity in 15 and 21 days are correlated to those decolorized results. The dead cell was also able to decolorize 67.3% brownish liquor and 25.1% black liquor after 15 days incubation as biosorption mechanism. This research described fungal potential in decolorization as the simple black liquor treatment technology and gave valuable information related to environmental friendly decolorization process.

Decision analysis for conservation breeding: Maximizing production for reintroduction of whooping cranes

USGS Publications Warehouse

Smith, Des H.V.; Converse, Sarah J.; Gibson, Keith; Moehrenschlager, Axel; Link, William A.; Olsen, Glenn H.; Maguire, Kelly

2011-01-01

Captive breeding is key to management of severely endangered species, but maximizing captive production can be challenging because of poor knowledge of species breeding biology and the complexity of evaluating different management options. In the face of uncertainty and complexity, decision-analytic approaches can be used to identify optimal management options for maximizing captive production. Building decision-analytic models requires iterations of model conception, data analysis, model building and evaluation, identification of remaining uncertainty, further research and monitoring to reduce uncertainty, and integration of new data into the model. We initiated such a process to maximize captive production of the whooping crane (Grus americana), the world's most endangered crane, which is managed through captive breeding and reintroduction. We collected 15 years of captive breeding data from 3 institutions and used Bayesian analysis and model selection to identify predictors of whooping crane hatching success. The strongest predictor, and that with clear management relevance, was incubation environment. The incubation period of whooping crane eggs is split across two environments: crane nests and artificial incubators. Although artificial incubators are useful for allowing breeding pairs to produce multiple clutches, our results indicate that crane incubation is most effective at promoting hatching success. Hatching probability increased the longer an egg spent in a crane nest, from 40% hatching probability for eggs receiving 1 day of crane incubation to 95% for those receiving 30 days (time incubated in each environment varied independently of total incubation period). Because birds will lay fewer eggs when they are incubating longer, a tradeoff exists between the number of clutches produced and egg hatching probability. We developed a decision-analytic model that estimated 16 to be the optimal number of days of crane incubation needed to maximize the number of offspring produced. These results show that using decision-analytic tools to account for uncertainty in captive breeding can improve the rate at which such programs contribute to wildlife reintroductions. 

Evaluation and simulation of nitrogen mineralization of paddy soils in Mollisols area of Northeast China under waterlogged incubation

PubMed Central

Zhang, Yuling; Xu, Wenjing; Duan, Pengpeng; Cong, Yaohui; An, Tingting; Yu, Na; Zou, Hongtao; Dang, Xiuli; An, Jing; Fan, Qingfeng; Zhang, Yulong

2017-01-01

Background Understanding the nitrogen (N) mineralization process and applying appropriate model simulation are key factors in evaluating N mineralization. However, there are few studies of the N mineralization characteristics of paddy soils in Mollisols area of Northeast China. Materials and methods The soils were sampled from the counties of Qingan and Huachuan, which were located in Mollisols area of Northeast China. The sample soil was incubated under waterlogged at 30°C in a controlled temperature cabinet for 161 days (a 2: 1 water: soil ratio was maintained during incubation). Three models, i.e. the single first-order kinetics model, the double first-order kinetics model and the mixed first-order and zero-order kinetics model were used to simulate the cumulative mineralised N (NH4+-N and TSN) in the laboratory and waterlogged incubation. Principal results During 161 days of waterlogged incubation, the average cumulative total soluble N (TSN), ammonium N (NH4+-N), and soluble organic N (SON) was 122.2 mg kg-1, 85.9 mg kg-1, and 36.3 mg kg-1, respectively. Cumulative NH4+-N was significantly (P < 0.05) positively correlated with organic carbon (OC), total N (TN), pH, and exchangeable calcium (Ca), and cumulative TSN was significantly (P < 0.05) positively correlated with OC, TN, and exchangeable Ca, but was not significantly (P > 0.05) correlated with C/N ratio, cation exchange capacity (CEC), extractable iron (Fe), clay, and sand. When the cumulative NH4+-N and TSN were simulated, the single first-order kinetics model provided the least accurate simulation. The parameter of the double first-order kinetics model also did not represent the actual data well, but the mixed first-order and zero-order kinetics model provided the most accurate simulation, as demonstrated by the estimated standard error, F statistic values, parameter accuracy, and fitting effect. Conclusions Overall, the results showed that SON was involved with N mineralization process, and the mixed first-order and zero-order kinetics model accurately simulates the N mineralization process of paddy soil in Mollisols area of Northeast China under waterlogged incubation. PMID:28170409

Evaluation and simulation of nitrogen mineralization of paddy soils in Mollisols area of Northeast China under waterlogged incubation.

PubMed

Zhang, Yuling; Xu, Wenjing; Duan, Pengpeng; Cong, Yaohui; An, Tingting; Yu, Na; Zou, Hongtao; Dang, Xiuli; An, Jing; Fan, Qingfeng; Zhang, Yulong

2017-01-01

Understanding the nitrogen (N) mineralization process and applying appropriate model simulation are key factors in evaluating N mineralization. However, there are few studies of the N mineralization characteristics of paddy soils in Mollisols area of Northeast China. The soils were sampled from the counties of Qingan and Huachuan, which were located in Mollisols area of Northeast China. The sample soil was incubated under waterlogged at 30°C in a controlled temperature cabinet for 161 days (a 2: 1 water: soil ratio was maintained during incubation). Three models, i.e. the single first-order kinetics model, the double first-order kinetics model and the mixed first-order and zero-order kinetics model were used to simulate the cumulative mineralised N (NH4+-N and TSN) in the laboratory and waterlogged incubation. During 161 days of waterlogged incubation, the average cumulative total soluble N (TSN), ammonium N (NH4+-N), and soluble organic N (SON) was 122.2 mg kg-1, 85.9 mg kg-1, and 36.3 mg kg-1, respectively. Cumulative NH4+-N was significantly (P < 0.05) positively correlated with organic carbon (OC), total N (TN), pH, and exchangeable calcium (Ca), and cumulative TSN was significantly (P < 0.05) positively correlated with OC, TN, and exchangeable Ca, but was not significantly (P > 0.05) correlated with C/N ratio, cation exchange capacity (CEC), extractable iron (Fe), clay, and sand. When the cumulative NH4+-N and TSN were simulated, the single first-order kinetics model provided the least accurate simulation. The parameter of the double first-order kinetics model also did not represent the actual data well, but the mixed first-order and zero-order kinetics model provided the most accurate simulation, as demonstrated by the estimated standard error, F statistic values, parameter accuracy, and fitting effect. Overall, the results showed that SON was involved with N mineralization process, and the mixed first-order and zero-order kinetics model accurately simulates the N mineralization process of paddy soil in Mollisols area of Northeast China under waterlogged incubation.

Evaluation of decontamination process of heart valve and artery tissues in European Homograft Bank (EHB): a retrospective study of 1,055 cases.

PubMed

Fan, Ye-Dong; Van Hoeck, Beatrice; Holovska, Vanda; Jashari, Ramadan

2012-06-01

To evaluate the efficiency of decontamination practice in European Homograft Bank (EHB), the data of the cardiovascular tissues received during recent 2 years were retrospectively analysed in this study. After initial assessment, the tissues were incubated in a 3-antibiotics' cocktail at 4°C for 20-48 h. The states of contamination were evaluated before and after incubation with the focus on the differences in donor type, tissue type, germ type and incubation time. Amongst 1,055 eligible tissues, 77.2% were hearts and 22.8% were arteries. 82.2% of the tissues were retrieved from the multi-organ donors (MOD), 15.4% from the recipients of heart transplantation (RHT) and 2.4% from the non-heart beating donors (NHBD). The initial contamination rate was 27.4% with a significantly higher incidence in arteries. The RHT tissues had the lowest contamination rate comparing to that of MOD and NHBD. Staphylococcus species was the major source of contamination. After antibiotic incubation, 76.8% of the contaminated tissues were disinfected, which was significantly higher for the hearts than the arteries. The RHT tissues had the highest decontamination rate than that of MOD and NHBD tissues. Propionibacterium acnes was detected in 48.1% of the remaining contaminated cases. The average incubation time of the Propionibacterium-positive tissues was significantly shorter than that of decontaminated tissues. In conclusion, the current decontamination protocol of EHB is sufficient for most of the initially contaminated bacteria, whereas it is inadequate for Propionibacterium acnes. This may be related to the slow-growing nature of this bacterium and thereby the relative shorter antibiotic incubation time.

Effect of sodium nitrite and sodium nitrate on botulinal toxin production and nitrosamine formation in wieners.

PubMed

Hustad, G O; Cerveny, J G; Trenk, H; Deibel, R H; Kautter, D A; Fazio, T; Johnston, R W; Kolari, O E

1973-07-01

Wieners were formulated and processed approximating commercial conditions as closely as possible. Twenty-four batches of product were made with the addition of six levels of sodium nitrite (0, 50, 100, 150, 200, and 300 mug/g), four levels of sodium nitrate (0, 50, 150, and 450 mug/g), and two levels of Clostridium botulinum (0 and 620 spores/g). After formulation, processing, and vacuum packaging, portions of each batch were incubated at 27 C or held for 21 days at 7 C followed by incubation at 27 C for 56 days. The latter storage condition approximated distribution of product through commercial channels and potential temperature abuse at the consumer level. Samples were analyzed for botulinal toxin, nitrite, and nitrate levels after 3, 7, 14, 21, 28, and 56 days of incubation. When nitrite was not added, toxic samples were detected after 14 days of incubation at 27 C. At the lowest level of nitrite added (50 mug/g), no toxic samples were observed until 56 days of incubation. Higher levels of nitrite completely inhibited toxin production throughout the incubation period. Nine uninoculated samples, representing various levels and combinations of nitrite and nitrate, were evaluated organoleptically. The flavor quality of wieners made with nitrite was judged significantly higher (P = 0.05) than of wieners made without nitrite. The nine samples were negative for 14 volatile nitrosamines at a sensitivity level of 10 ng/g. The results indicated that nitrite effectively inhibited botulinal toxin formation at commercially employed levels in wieners and that detectable quantities of nitrosamines were not produced during preparation and processing of the product for consumption.

Effect of Sodium Nitrite and Sodium Nitrate on Botulinal Toxin Production and Nitrosamine Formation in Wieners

PubMed Central

Hustad, Gerald O.; Cerveny, John G.; Trenk, Hugh; Deibel, Robert H.; Kautter, Donald A.; Fazio, Thomas; Johnston, Ralph W.; Kolari, Olaf E.

1973-01-01

Wieners were formulated and processed approximating commercial conditions as closely as possible. Twenty-four batches of product were made with the addition of six levels of sodium nitrite (0, 50, 100, 150, 200, and 300 μg/g), four levels of sodium nitrate (0, 50, 150, and 450 μg/g), and two levels of Clostridium botulinum (0 and 620 spores/g). After formulation, processing, and vacuum packaging, portions of each batch were incubated at 27 C or held for 21 days at 7 C followed by incubation at 27 C for 56 days. The latter storage condition approximated distribution of product through commercial channels and potential temperature abuse at the consumer level. Samples were analyzed for botulinal toxin, nitrite, and nitrate levels after 3, 7, 14, 21, 28, and 56 days of incubation. When nitrite was not added, toxic samples were detected after 14 days of incubation at 27 C. At the lowest level of nitrite added (50 μg/g), no toxic samples were observed until 56 days of incubation. Higher levels of nitrite completely inhibited toxin production throughout the incubation period. Nine uninoculated samples, representing various levels and combinations of nitrite and nitrate, were evaluated organoleptically. The flavor quality of wieners made with nitrite was judged significantly higher (P = 0.05) than of wieners made without nitrite. The nine samples were negative for 14 volatile nitrosamines at a sensitivity level of 10 ng/g. The results indicated that nitrite effectively inhibited botulinal toxin formation at commercially employed levels in wieners and that detectable quantities of nitrosamines were not produced during preparation and processing of the product for consumption. PMID:4580194

21 CFR 113.83 - Establishing scheduled processes.

Code of Federal Regulations, 2010 CFR

2010-04-01

... shall be performed according to procedures recognized by competent processing authorities. If incubation... actual commercial production runs during the period of instituting the process. The incubation tests for... of the process and associated incubation tests shall be prepared and shall be permanently retained by...

21 CFR 113.83 - Establishing scheduled processes.

Code of Federal Regulations, 2011 CFR

2011-04-01

... procedures recognized by competent processing authorities. If incubation tests are necessary for process... the period of instituting the process. The incubation tests for confirmation of the scheduled... and associated incubation tests shall be prepared and shall be permanently retained by the person or...

9 CFR 355.25 - Canning with heat processing and hermetically sealed containers; closures; code marking; heat...

Code of Federal Regulations, 2013 CFR

2013-01-01

... hermetically sealed containers; closures; code marking; heat processing; incubation. 355.25 Section 355.25... processing and hermetically sealed containers; closures; code marking; heat processing; incubation. (a... storage and transportation as evidenced by the incubation test. (h) Lots of canned products shall be...

9 CFR 355.25 - Canning with heat processing and hermetically sealed containers; closures; code marking; heat...

Code of Federal Regulations, 2012 CFR

2012-01-01

... hermetically sealed containers; closures; code marking; heat processing; incubation. 355.25 Section 355.25... processing and hermetically sealed containers; closures; code marking; heat processing; incubation. (a... storage and transportation as evidenced by the incubation test. (h) Lots of canned products shall be...

9 CFR 355.25 - Canning with heat processing and hermetically sealed containers; closures; code marking; heat...

Code of Federal Regulations, 2011 CFR

2011-01-01

... hermetically sealed containers; closures; code marking; heat processing; incubation. 355.25 Section 355.25... processing and hermetically sealed containers; closures; code marking; heat processing; incubation. (a... storage and transportation as evidenced by the incubation test. (h) Lots of canned products shall be...

9 CFR 355.25 - Canning with heat processing and hermetically sealed containers; closures; code marking; heat...

Code of Federal Regulations, 2014 CFR

2014-01-01

... hermetically sealed containers; closures; code marking; heat processing; incubation. 355.25 Section 355.25... processing and hermetically sealed containers; closures; code marking; heat processing; incubation. (a... storage and transportation as evidenced by the incubation test. (h) Lots of canned products shall be...

9 CFR 355.25 - Canning with heat processing and hermetically sealed containers; closures; code marking; heat...

Code of Federal Regulations, 2010 CFR

2010-01-01

... hermetically sealed containers; closures; code marking; heat processing; incubation. 355.25 Section 355.25... processing and hermetically sealed containers; closures; code marking; heat processing; incubation. (a... storage and transportation as evidenced by the incubation test. (h) Lots of canned products shall be...

An exposure-response analysis based on rifampin suggests CYP3A4 induction is driven by AUC: an in vitro investigation.

PubMed

Chang, Cheng; Yang, Xin; Fahmi, Odette A; Riccardi, Keith A; Di, Li; Obach, R Scott

2017-08-01

1. Induction is an important mechanism contributing to drug-drug interactions. It is most commonly evaluated in the human hepatocyte assay over 48-h or 72-h incubation period. However, whether the overall exposure (i.e. Area Under the Curve (AUC) or C ave ) or maximum exposure (i.e. C max ) of the inducer is responsible for the magnitude of subsequent induction has not been thoroughly investigated. Additionally, in vitro induction assays are typically treated as static systems, which could lead to inaccurate induction potency estimation. Hence, European Medicines Agency (EMA) guidance now specifies quantitation of drug levels in the incubation. 2. This work treated the typical in vitro evaluation of rifampin induction as an in vivo system by generating various target engagement profiles, measuring free rifampin concentration over 3 d of incubation and evaluating the impact of these factors on final induction response. 3. This rifampin-based analysis demonstrates that the induction process is driven by time-averaged target engagement (i.e. AUC-driven). Additionally, depletion of rifampin in the incubation medium over 3 d as well as non-specific/specific binding were observed. 4. These findings should help aid the discovery of clinical candidates with minimal induction liability and further expand our knowledge in the quantitative translatability of in vitro induction assays.

Formation of Humic Substances in Weathered MSWI Bottom Ash

PubMed Central

Zhang, Haixia; Shimaoka, Takayuki

2013-01-01

The study aimed at evaluating the humic substances (HSs) content from municipal solid waste incinerator (MSWI) bottom ash and its variation with time and the effect of temperature on HSs formation. The process suggested by IHSS was applied to extract HSs from two different bottom ash samples, and the extracted efficiency with NaOH and Na4P2O7 was compared. MSWI bottom ash samples were incubated at 37°C and 50°C for 1 year. HSs and nonhumic substances were extracted from the bottom ash sample with different incubated period by 0.1 M NaOH/Na4P2O7. Results show that the rate of humic acid formation increased originally with incubation time, reached a maximum at 12th week under 37°C and at 18th week under 50°C, and then decreased with time. More humic acid in MSWI bottom ash was formed under 50°C incubated condition compared with that incubated under 37°C. Also, the elemental compositions of HSs extracted from bottom ash are reported. PMID:23844394

Factors affecting incubation patterns and sex roles of black oystercatchers in Alaska

USGS Publications Warehouse

Spiegel, Caleb S.; Haig, Susan M.; Goldstein, Michael I.; Huso, Manuela M. P.

2012-01-01

Studies examining the effects of human disturbance on avian parental behavior and reproductive success are fundamental to bird conservation. However, many such studies fail to also consider the influence of natural threats, a variable environment, and parental roles. Our work examines interactive relationships of cyclical (time of day, tide, temperature, seasonality) and stochastic (natural/human disturbance) processes with incubation patterns (attendance, bout lengths, recess rates) of the Black Oystercatcher (Haematopus bachmani), a shorebird of conservation concern. We used 24-hour-per-day video monitoring of 13 molecularly-sexed breeding pairs to systematically examine incubation, revealing previously undocumented information that may inform conservation practices for the genus. Seven of 22 video-monitored nests failed, primarily from egg depredation by nocturnally-active mammals. Analyses of 3177 hrs of video footage indicated a near doubling of incubation bout lengths at night, corresponding to the increased risk of nighttime egg predation. Females had higher overall nest attendance (54% vs. 42%) and longer mean incubation bout lengths than males (88 min vs. 73 min). Uninterrupted incubation bouts were over twice as long as bouts interrupted by disturbance. Incubating males departed nests substantially more frequently due to nest-area disturbances than females in one, but not both, years of our study. Our findings suggest that sexes exhibit different, but complimentary, incubation patterns, facilitating efficient egg care in a dynamic environment with several nest threats. We emphasize the importance of considering natural influences when evaluating human threats to shorebird reproductive behavior and success.

Aligning business strategy of incubator center and tenants

NASA Astrophysics Data System (ADS)

Prasetyawan, Yudha; Agustiani, Elly; Jumayla, Sari

2017-06-01

Incubator center is developed to help a particular group of small business players to achieve the expected business growth. In this center, business players often called as tenants will get assistances in pertaining with space, professional network, marketing, investment or funding, and training to improve their business capability. There are three types of incubator center, namely universities that help their alumni or business people in their surrounded area, company that supports small business as the corporate social responsibility, and independent organizations that have specialties in the business development. Some might success in increasing the capacity of the tenants, while other can have difficulties to increase the simplest business capability, e.g., to define the production cost to measure the profit. This study was intended to propose a model to align the business strategy between incubator center and its tenants. The sales and profit growth are the main priorities for the tenants together with their business capability and sustainability. The proposed alignment model provides measurement tools that link the motivation of tenants for joining the incubation process with the mission of incubator center. The linkage covered the key performance indicators (KPI), steps to achieve the target and evaluation tools to improve the current handicaps. An experiment on 4 (four) diverse business fields of the tenants of an incubator center was performed to test the model. As a result, the increase of KPI of incubator center will simultaneously yield a higher value of the tenants' sales.

Monte-Carlo simulation of defect-cluster nucleation in metals during irradiation

NASA Astrophysics Data System (ADS)

Nakasuji, Toshiki; Morishita, Kazunori; Ruan, Xiaoyong

2017-02-01

A multiscale modeling approach was applied to investigate the nucleation process of CRPs (copper rich precipitates, i.e., copper-vacancy clusters) in α-Fe containing 1 at.% Cu during irradiation. Monte-Carlo simulations were performed to investigate the nucleation process, with the rate theory equation analysis to evaluate the concentration of displacement defects, along with the molecular dynamics technique to know CRP thermal stabilities in advance. Our MC simulations showed that there is long incubation period at first, followed by a rapid growth of CRPs. The incubation period depends on irradiation conditions such as the damage rate and temperature. CRP's composition during nucleation varies with time. The copper content of CRPs shows relatively rich at first, and then becomes poorer as the precipitate size increases. A widely-accepted model of CRP nucleation process is finally proposed.

9 CFR 381.309 - Finished product inspection.

Code of Federal Regulations, 2012 CFR

2012-01-01

... alternative documented procedures for handling process deviations. (1) Incubation of shelf stable canned product—(i) Incubator. The establishment shall provide incubation facilities which include an accurate... the facility. The Program is responsible for the security of the incubator. (ii) Incubation...

9 CFR 381.309 - Finished product inspection.

Code of Federal Regulations, 2013 CFR

2013-01-01

... alternative documented procedures for handling process deviations. (1) Incubation of shelf stable canned product—(i) Incubator. The establishment shall provide incubation facilities which include an accurate... the facility. The Program is responsible for the security of the incubator. (ii) Incubation...

9 CFR 381.309 - Finished product inspection.

Code of Federal Regulations, 2014 CFR

2014-01-01

... alternative documented procedures for handling process deviations. (1) Incubation of shelf stable canned product—(i) Incubator. The establishment shall provide incubation facilities which include an accurate... the facility. The Program is responsible for the security of the incubator. (ii) Incubation...

Rapid development of xylanase assay conditions using Taguchi methodology.

PubMed

Prasad Uday, Uma Shankar; Bandyopadhyay, Tarun Kanti; Bhunia, Biswanath

2016-11-01

The present investigation is mainly concerned with the rapid development of extracellular xylanase assay conditions by using Taguchi methodology. The extracellular xylanase was produced from Aspergillus niger (KP874102.1), a new strain isolated from a soil sample of the Baramura forest, Tripura West, India. Four physical parameters including temperature, pH, buffer concentration and incubation time were considered as key factors for xylanase activity and were optimized using Taguchi robust design methodology for enhanced xylanase activity. The main effect, interaction effects and optimal levels of the process factors were determined using signal-to-noise (S/N) ratio. The Taguchi method recommends the use of S/N ratio to measure quality characteristics. Based on analysis of the S/N ratio, optimal levels of the process factors were determined. Analysis of variance (ANOVA) was performed to evaluate statistically significant process factors. ANOVA results showed that temperature contributed the maximum impact (62.58%) on xylanase activity, followed by pH (22.69%), buffer concentration (9.55%) and incubation time (5.16%). Predicted results showed that enhanced xylanase activity (81.47%) can be achieved with pH 2, temperature 50°C, buffer concentration 50 Mm and incubation time 10 min.

Ambient-temperature incubation for the field detection of Escherichia coli in drinking water.

PubMed

Brown, J; Stauber, C; Murphy, J L; Khan, A; Mu, T; Elliott, M; Sobsey, M D

2011-04-01

 Escherichia coli is the pre-eminent microbiological indicator used to assess safety of drinking water globally. The cost and equipment requirements for processing samples by standard methods may limit the scale of water quality testing in technologically less developed countries and other resource-limited settings, however. We evaluate here the use of ambient-temperature incubation in detection of E. coli in drinking water samples as a potential cost-saving and convenience measure with applications in regions with high (>25°C) mean ambient temperatures.   This study includes data from three separate water quality assessments: two in Cambodia and one in the Dominican Republic. Field samples of household drinking water were processed in duplicate by membrane filtration (Cambodia), Petrifilm™ (Cambodia) or Colilert® (Dominican Republic) on selective media at both standard incubation temperature (35–37°C) and ambient temperature, using up to three dilutions and three replicates at each dilution. Matched sample sets were well correlated with 80% of samples (n = 1037) within risk-based microbial count strata (E. coli CFU 100 ml−1 counts of <1, 1–10, 11–100, 101–1000, >1000), and a pooled coefficient of variation of 17% (95% CI 15–20%) for paired sample sets across all methods.   These results suggest that ambient-temperature incubation of E. coli in at least some settings may yield sufficiently robust data for water safety monitoring where laboratory or incubator access is limited.

Efficacy of decontamination protocol by antimicrobial treatment in Iranian Tissue Bank (ITB).

PubMed

Dashti-Khavidaki, Simin; Dabardani, Fariba; Mahdavi-Mazdeh, Mitra; Ravanasa, Elham; Hosseini, Seyed Kazem

2015-09-01

Iranian Tissue Bank established in 1994 provides soft tissues for implantation in Iran. This study was designed to evaluate the efficacy of decontamination process of cardiac and soft tissues in Iranian Tissue Bank. In this bank after initial assessments, the tissues were incubated in a 5-antibiotic cocktail at room temperature for 24 h and then at 4 °C for 14 days. Contamination status was compared before and after antibiotic cocktail incubation. Of 3,315 assessed tissues, 1,057 were pericardia, 1,051 were fascia and 1,207 were other soft tissues including tibialis and aorta. The initial contamination rate was 36.86%. Pericardia showed the highest contamination rate. Klebsiella species was the most prevalent organism causing contamination. Decontamination rate after antibiotic incubation was 86.91% with the highest successful decontamination rate for fascia tissue. Klebsiella species was the major source of contamination in tissues that remained contaminated after antibiotic incubation. This may be due to resistance of this organism to applied antibiotics in the decontamination cocktail possibly due to a negative drug interaction between aminoglycoside and penicillin derivatives in this antibiotic cocktail. In conclusion collected data shows comparable efficacy of the decontamination process that is used in ITB compared with homograft banks of other countries.

Impact of prolonged oocyte incubation time before vitrification on oocyte survival, embryo formation, and embryo quality in mice.

PubMed

Karami, Azade; Bakhtiari, Mitra; Azadbakht, Mehri; Ghorbani, Rostam; Khazaei, Mozafar; Rezaei, Mansour

2017-06-01

Oocyte incubation time before freezing is one of the factors affecting oocyte vitrification. In the assisted reproductive technology (ART) clinics, it is sometimes decided to perform oocyte vitrification after a long period of incubation time due to various conditions, such as inability to collect semen samples, unsuccessful urological interventions (PESA, TESE, etc.), or unexpected conditions. A time factor of up to 6 h has been studied in the available reports. Therefore, this study was designed to evaluate oocyte incubation time before freezing at 0, 6, 12, 18, and 24 h after retrieval. Metaphase II (MII) oocytes were obtained from NMRI female mice after being randomly divided into the five groups of 0, 6, 12, 18, and 24 h of freezing via hormonal stimulation following retrieval and entered into the vitrification-warming process. The thawed oocytes were evaluated according to the survival criteria and then inseminated with the sperms of male mice for in vitro fertilization. The next day, the embryo formation rate and embryo quality were assessed. Our results demonstrated that even after 24 h of incubation, the survival rate of oocytes was 51.35% with the embryo formation rate of 73.21%. However, the survival and embryo formation rates significantly decreased within 12, 18, and 24 h after retrieval compared to the groups vitrified at 0 h. The embryo quality was significantly reduced by vitrification at 0 to 24 h after retrieval. According to our data, although a prolonged incubation time before freezing reduced the survival rate, there was still a chance for oocytes to stay alive with acceptable embryo formation and quality rates after vitrification warming of oocytes.

Best Practices for In-Situ Sediment-Water Incubations with Benthic Landers

NASA Astrophysics Data System (ADS)

Tengberg, Anders; Kononets, Mikhail; Hall, Per; Nilsson, Madeleine; Ekeroth, Nils

2017-04-01

Biological, chemical, physical and geological processes that take place at the seafloor are crucial in influencing and regulating many aquatic environments. One method to estimate exchange rates, fluxes, between the sediment and the overlying water is in-situ sediment-water incubations using autonomous chamber landers. As for all field sampling and measurements best practices methods are needed to obtain high quality data. With experiences form many years usage of the Gothenburg autonomous bottom lander systems this presentation will describe some of the experimental work that has been done with focus on quality control and data evaluation methods.

The impact of incubating the throat culture for 72 h on the diagnosis of group A beta-hemolytic streptococci.

PubMed

Kocoglu, Esra; Karabay, Oguz; Yilmaz, Fahrettin; Ekerbicer, Hasan

2006-09-01

The accurate detection and treatment of group A beta-hemolytic streptococcal pharyngitis is important to prevent the potential sequels. Throat culture remains the gold standard for the diagnosis of streptococcal pharyngitis. Many authors recommended 48 h incubation time for the isolation of group A Streptococcus (GAS). In this study the diagnostic efficacy of prolonging the incubation time of throat cultures to 72 h for isolation of GAS was researched. Throat swab specimens were inoculated to 5% sheep blood agar plate and incubated at 37 degrees C, and evaluated at the 24th hour. GAS negative plates were incubated again for another 24h and read at the 48th hour. Negative plates at 48 h were incubated again, read at the 72nd hour, and positive results of all measurements were evaluated. A total of 367 cultures were examined. At the 24th hour evaluation 34 (9.3%) cultures were positive for GAS. Evaluation of 333 cultures that were found to be negative in the first 24th hour evaluation revealed 44 (13.6%) positive cultures at 48th hour evaluation. Out of 289 cultures that were negative during the 48th hour evaluation, an additional 13 (4.7%) cultures were found to be positive during the 72nd hour evaluation. The agreement between 24th-48th and 24th and 72nd were both moderate (kappa 0.54 and 0. 46, respectively), and between 48th and 72nd hour was very good (kappa: 0.90). Throat cultures that are negative for GAS at 24h should be incubated for second day. We conclude that, to confirm the throat cultures GAS negative, results of throat cultures need to be incubated for 72 h, since 72nd hour evaluations revealed additional 5% positive results.

Screening and optimization of pectin lyase and polygalacturonase activity from ginseng pathogen Cylindrocarpon Destructans

PubMed Central

Sathiyaraj, Gayathri; Srinivasan, Sathiyaraj; Kim, Ho-Bin; Subramaniyam, Sathiyamoorthy; Lee, Ok Ran; Kim, Yeon-Ju; Yang, Deok Chun

2011-01-01

Cylindrocarpon destructans isolated from ginseng field was found to produce pectinolytic enzymes. A Taguchi’s orthogonal array experimental design was applied to optimize the preliminary production of polygalacturonase (PG) and pectin lyase (PL) using submerged culture condition. This method was applied to evaluate the significant parameters for the production of enzymes. The process variables were pH, pectin concentration, incubation time and temperature. Optimization of process parameters resulted in high levels of enzyme (PG and PL) production after ten days of incubation at a pH of 5.0 at 25°C in the presence of 1.5% pectin. Among different nitrogen sources, urea and peptone showed high production of PG and PL, respectively. The enzyme production and mycelial growth seems to have direct influence on the culture conditions; therefore, at stationary state high enzyme production and mycelial growth were obtained than agitation state. Along with this, optimization of enzyme activity was also determined using various physiological parameters like, temperature, incubation time and pH. Taguchi’s data was also analyzed using one step ANOVA statistical method. PMID:24031695

Effect of soil organic matter on antimony bioavailability after the remediation process.

PubMed

Nakamaru, Yasuo Mitsui; Martín Peinado, Francisco José

2017-09-01

We evaluated the long-term (18 year) and short-term (4 weeks) changes of Sb in contaminated soil with SOM increase under remediation process. In the Aznalcóllar mine accident (1998) contaminated area, the remediation measurement implemented the Guadiamar Green Corridor, where residual pollution is still detected. Soils of the re-vegetated area (O2) with high pH and high SOM content, moderately re-vegetated area (O1) and unvegetated area (C) were sampled. Soil pH, CEC, SOM amount and soil Sb forms were evaluated. Soil Sb was measured as total, soluble, exchangeable, EDTA extractable, acid oxalate extractable, and pyro-phosphate extractable fractions. Further, the short-term effect of artificial organic matter addition was also evaluated with incubation study by adding compost to the sampled soil from C, O1 and O2 areas. After 4 weeks of incubation, soil chemical properties and Sb forms were evaluated. In re-vegetated area (O2), soil total Sb was two times lower than in unvegetated area (C); however, soluble, exchangeable, and EDTA extractable Sb were 2-8 times higher. The mobile/bioavailable Sb increase was also observed after 4 weeks of incubation with the addition of compost. Soluble, exchangeable, and EDTA extractable Sb was increased 2-4 times by compost addition. By the linear regression analysis, the significantly related factors for soluble, exchangeable, and EDTA extractable Sb values were pH, CEC, and SOM, respectively. Soluble Sb increase was mainly related to pH rise. Exchangeable Sb should be bound by SOM-metal complex and increased with CEC. EDTA extractable fraction should be increased with increase of SOM as SOM-Fe associated Sb complex. From these results, it was shown that increase of SOM under natural conditions or application of organic amendment under remediation process should increase availability of Sb to plants. Copyright © 2017 Elsevier Ltd. All rights reserved.

Tenant Recruitment and Support Processes in Sustainability-Profiled Business Incubators

ERIC Educational Resources Information Center

Bank, Natasha; Kanda, Wisdom

2016-01-01

Recruitment and support processes in sustainability-profiled incubators have received little research attention. The article addresses this knowledge gap in an empirical investigation of three sustainability-oriented incubators in Sweden, Finland and Germany. The data are based on interviews with managers, stakeholders and tenants in Green Tech…

Effect of eggshell temperature and oxygen concentration during incubation on the developmental and physiological status of broiler hatchlings in the perinatal period.

PubMed

Molenaar, R; van den Anker, I; Meijerhof, R; Kemp, B; van den Brand, H

2011-06-01

This study evaluated the influence of incubation conditions on the developmental and physiological status of birds in the perinatal period, which spans the end of incubation until the early posthatch period. Embryos were incubated at a normal (37.8°C) or high (38.9°C) eggshell temperature (EST) and a low (17%), normal (21%), or high (25%) O(2) concentration from d 7 until 19 of incubation. After d 19 of incubation, EST was maintained, but O(2) concentrations were 21% for all embryos. Body and organ weights, and hepatic glycogen levels were measured at d 18 of incubation and at 12 and 48 h after emergence from the eggshell. In addition, blood metabolites were measured at 12 and 48 h after emergence from the eggshell. Embryos incubated at a high EST and low O(2) concentration had the highest mortality in the last week of incubation, which may be related to their low yolk-free body mass (YFBM) or a reduced nutrient availability for hatching (i.e., hepatic glycogen). High EST, compared with normal EST, decreased YFBM. This may be due to the shorter incubation duration of 8 h, the lower weight of supply organs (i.e., heart and lung), or a lack of glucose precursors. Because of this lack of glucose precursors, embryos incubated at high EST may have used proteins for energy production instead of for body development at the end of incubation. The YFBM at d 18 of incubation increased with an increase in O(2) concentration. However, differences between the normal and high O(2) concentration disappeared at 12 and 48 h after emergence, possibly because the high O(2) concentration had difficulties adapting to lower O(2) concentrations in the perinatal period. Blood metabolites and hepatic glycogen were comparable among O(2) concentrations, indicating that the physiological status at hatch may be related to the environment that the embryo experienced during the hatching process. In conclusion, EST and O(2) concentration differentially influence the developmental and physiological status of broilers during the perinatal period.

Multicenter Study on Incubation Conditions for Environmental Monitoring and Aseptic Process Simulation.

PubMed

Guinet, Roland; Berthoumieu, Nicole; Dutot, Philippe; Triquet, Julien; Ratajczak, Medhi; Thibaudon, Michel; Bechaud, Philippe; Arliaud, Christophe; Miclet, Edith; Giordano, Florine; Larcon, Marjorie; Arthaud, Catherine

Environmental monitoring and aseptic process simulations represent an integral part of the microbiological quality control system of sterile pharmaceutical products manufacturing operations. However, guidance documents and manufacturers practices differ regarding recommendations for incubation time and incubation temperature, and, consequently, the environmental monitoring and aseptic process simulation incubation strategy should be supported by validation data. To avoid any bias coming from in vitro studies or from single-site manufacturing in situ studies, we performed a collaborative study at four manufacturing sites with four samples at each location. The environmental monitoring study was performed with tryptic soy agar settle plates and contact plates, and the aseptic process simulation study was performed with tryptic soy broth and thioglycolate broth. The highest recovery rate was obtained with settle plates (97.7%) followed by contact plates (65.4%) and was less than 20% for liquid media (tryptic soy broth 19% and thioglycolate broth 17%). Gram-positive cocci and non-spore-forming Gram-positive rods were largely predominant with more than 95% of growth and recovered best at 32.5 °C. The highest recovery of molds was obtained at 22.5 °C alone or as the first incubation temperature. Strict anaerobes were not recovered. At the end of the five days of incubation no significant statistical difference was obtained between the four conditions. Based on these data a single incubation temperature at 32.5 °C could be recommended for these four manufacturing sites for both environmental monitoring and aseptic process simulation, and a second plate could be used, periodically incubated at 22.5 °C. Similar studies should be considered for all manufacturing facilities in order to determine the optimal incubation temperature regime for both viable environmental monitoring and aseptic process simulation. Microbiological environmental monitoring and aseptic process simulation confirm that pharmaceutical cleanrooms are in an appropriate hygienic condition for manufacturing of sterile drug products. Guidance documents from different health authorities or expert groups differ regarding recommendation of the applied incubation time and incubation temperature, leading to variable manufacturers practices. Some recent publications have demonstrated that laboratory studies are not relevant to determine the best incubation regime and that in situ manufacturing site studies should be used. To solve any possible bias coming from laboratory studies or single-site in situ studies, we conducted a multicenter study at four manufacturing sites with a significant amount of real environmental monitoring samples collected directly from the environment in pharmaceutical production during manufacturing operations with four solid and liquid nutrient media. These samples were then incubated under four different conditions suggested in the guidance documents. We believe that the results of our multicenter study confirming recent other single-site in situ studies could be the basis of the strategy to determine the best incubation regime for both viable environmental monitoring and aseptic process simulation in any manufacturing facility. © PDA, Inc. 2017.

Short mechanical biological treatment of municipal solid waste allows landfill impact reduction saving waste energy content.

PubMed

Scaglia, Barbara; Salati, Silvia; Di Gregorio, Alessandra; Carrera, Alberto; Tambone, Fulvia; Adani, Fabrizio

2013-09-01

The aim of this work was to evaluate the effects of full scale MBT process (28 d) in removing inhibition condition for successive biogas (ABP) production in landfill and in reducing total waste impact. For this purpose the organic fraction of MSW was treated in a full-scale MBT plant and successively incubated vs. untreated waste, in simulated landfills for one year. Results showed that untreated landfilled-waste gave a total ABP reduction that was null. On the contrary MBT process reduced ABP of 44%, but successive incubation for one year in landfill gave a total ABP reduction of 86%. This ABP reduction corresponded to a MBT process of 22 weeks length, according to the predictive regression developed for ABP reduction vs. MBT-time. Therefore short MBT allowed reducing landfill impact, preserving energy content (ABP) to be produced successively by bioreactor technology since pre-treatment avoided process inhibition because of partial waste biostabilization. Copyright © 2013 Elsevier Ltd. All rights reserved.

Creativity-the unconscious foundations of the incubation period.

PubMed

Ritter, Simone M; Dijksterhuis, Ap

2014-01-01

Creativity is one of the most important assets we have to navigate through the fast changing world of the 21st century. Anecdotal accounts of creative individuals suggest that oftentimes, creative discoveries result from a process whereby initial conscious thought is followed by a period during which one refrains from task-related conscious thought. For example, one may spend an embarrassing amount of time thinking about a problem when the solution suddenly pops into consciousness while taking a shower. Not only creative individuals but also traditional theories of creativity have put a lot of emphasis on this incubation stage in creative thinking. The aim of the present article is twofold. First, an overview of the domain of incubation and creativity is provided by reviewing and discussing studies on incubation, mind-wandering, and sleep. Second, the causes of incubation effects are discussed. Previously, little attention has been paid to the causes of incubation effects and most findings do not really speak to whether the effects should be explained by unconscious processes or merely by consequences of a period of distraction. In the latter case, there is no need to assume active unconscious processes. The findings discussed in the current article support the idea that it is not merely the absence of conscious thought that drives incubation effects, but that during an incubation period unconscious processes contribute to creative thinking. Finally, practical implications and directions for future research will be discussed.

Participation of the sperm proteasome during in vitro fertilisation and the acrosome reaction in cattle.

PubMed

Sánchez, R; Deppe, M; Schulz, M; Bravo, P; Villegas, J; Morales, P; Risopatrón, J

2011-04-01

In this work, we have investigated the role of the bovine sperm proteasome during in vitro fertilisation (IVF) and the acrosome reaction (AR). Motile spermatozoa, obtained by a swim-up method in Sperm-Talp medium, were capacitated for 3.5 h and incubated in the presence or absence of the specific proteasome inhibitor epoxomicin for 30 and 60 min. Then, the spermatozoa were co-incubated with mature bovine cumulus oocytes and after 48 h the cleavage rate of inseminated oocytes was evaluated. In addition, we evaluated the participation of the sperm proteasome during the progesterone-induced AR. Capacitated spermatozoa were incubated for 30 min with or without epoxomicin, then progesterone was added and the ARs were evaluated using the dual fluorescent staining technique 'Hoechst and chlortetracycline'. The results indicate that the proteasome inhibitor decreased the cleavage rate of oocytes inseminated with treated spermatozoa. In addition, acrosomal exocytosis levels were statistically significantly higher in the samples treated with the AR inducer progesterone than in control samples in the absence of the inducer. However, the progesterone-induced AR was significantly reduced by previous treatment of the spermatozoa with epoxomicin (P < 0.001). These observations indicate that the bovine sperm proteasome participates in the IVF and AR processes. © 2011 Blackwell Verlag GmbH.

Effects of Alcohol on Scientific Thought during the Incubation Phase of the Creative Process.

ERIC Educational Resources Information Center

Norlander, Torsten; Gustafson, Roland

1996-01-01

This study of 60 Swedish university students examined whether or not acute alcohol intoxication inhibits or facilitates the incubation phase of the creative process. Results found that the participants in the alcohol group produced significantly more incubations in their diaries and significantly higher levels of originality. (Author/CR)

Industrial yogurt manufacture: monitoring of fermentation process and improvement of final product quality.

PubMed

Soukoulis, C; Panagiotidis, P; Koureli, R; Tzia, C

2007-06-01

Lactic acid fermentation during the production of skim milk and whole fat set-style yogurt was continuously monitored by measuring pH. The modified Gompertz model was successfully applied to describe the pH decline and viscosity development during the fermentation process. The viscosity and incubation time data were also fitted to linear models against ln(pH). The investigation of the yogurt quality improvement practices included 2 different heat treatments (80 degrees C for 30 min and 95 degrees C for 10 min), 3 milk protein fortifying agents (skim milk powder, whey powder, and milk protein concentrate) added at 2.0%, and 4 hydrocolloids (kappa-carrageenan, xanthan, guar gum, and pectin) added at 0.01% to whole fat and skim yogurts. Heat treatment significantly affected viscosity and acetaldehyde development without influencing incubation time and acidity. The addition of whey powder shortened the incubation time but had a detrimental effect on consistency, firmness, and overall acceptance of yogurts. On the other hand, addition of skim milk powder improved the textural quality and decreased the vulnerability of yogurts to syneresis. Anionic stabilizers (kappa-carrageenan and pectin) had a poor effect on the texture and palatability of yogurts. However, neutral gums (xanthan and guar gum) improved texture and prevented the wheying-off defect. Skim milk yogurts exhibited longer incubation times and higher viscosities, whereas they were rated higher during sensory evaluation than whole fat yogurts.

Methanogenic and Sulfate-Reducing Activities in a Hypersaline Microbial Mat and Associated Microbial Diversity.

PubMed

Cadena, Santiago; García-Maldonado, José Q; López-Lozano, Nguyen E; Cervantes, Francisco J

2018-05-01

Methanogenesis and sulfate reduction are important microbial processes in hypersaline environments. However, key aspects determining substrate competition between these microbial processes have not been well documented. We evaluated competitive and non-competitive substrates for stimulation of both processes through microcosm experiments of hypersaline microbial mat samples from Guerrero Negro, Baja California Sur, Mexico, and we assessed the effect of these substrates on the microbial community composition. Methylotrophic methanogenesis evidenced by sequences belonging to methanogens of the family Methanosarcinaceae was found as the dominant methanogenic pathway in the studied hypersaline microbial mat. Nevertheless, our results showed that incubations supplemented with acetate and lactate, performed in absence of sulfate, also produced methane after 40 days of incubation, apparently driven by hydrogenotrophic methanogens affiliated to the family Methanomicrobiaceae. Sulfate reduction was mainly stimulated by addition of acetate and lactate; however, after 40 days of incubation, an increase of the H 2 S concentrations in microcosms amended with trimethylamine and methanol was also observed, suggesting that these substrates are putatively used for sulfate reduction. Moreover, 16S rRNA gene sequencing analysis showed remarkable differences in the microbial community composition among experimental treatments. In the analyzed sample amended with acetate, sulfate-reducing bacteria (SRB) belonging to the family Desulfobacteraceae were dominant, while members of Desulfohalobiaceae, Desulfomicrobiaceae, and Desulfovibrionaceae were found in the incubation with lactate. Additionally, we detected an unexpected high abundance of unclassified Hydrogenedentes (near 25%) in almost all the experimental treatments. This study contributes to better understand methanogenic and sulfate-reducing activities, which play an important role in the functioning of hypersaline environments.

Creativity—the unconscious foundations of the incubation period

PubMed Central

Ritter, Simone M.; Dijksterhuis, Ap

2014-01-01

Creativity is one of the most important assets we have to navigate through the fast changing world of the 21st century. Anecdotal accounts of creative individuals suggest that oftentimes, creative discoveries result from a process whereby initial conscious thought is followed by a period during which one refrains from task-related conscious thought. For example, one may spend an embarrassing amount of time thinking about a problem when the solution suddenly pops into consciousness while taking a shower. Not only creative individuals but also traditional theories of creativity have put a lot of emphasis on this incubation stage in creative thinking. The aim of the present article is twofold. First, an overview of the domain of incubation and creativity is provided by reviewing and discussing studies on incubation, mind-wandering, and sleep. Second, the causes of incubation effects are discussed. Previously, little attention has been paid to the causes of incubation effects and most findings do not really speak to whether the effects should be explained by unconscious processes or merely by consequences of a period of distraction. In the latter case, there is no need to assume active unconscious processes. The findings discussed in the current article support the idea that it is not merely the absence of conscious thought that drives incubation effects, but that during an incubation period unconscious processes contribute to creative thinking. Finally, practical implications and directions for future research will be discussed. PMID:24782742

Tying Theory To Practice: Cognitive Aspects of Computer Interaction in the Design Process.

ERIC Educational Resources Information Center

Mikovec, Amy E.; Dake, Dennis M.

The new medium of computer-aided design requires changes to the creative problem-solving methodologies typically employed in the development of new visual designs. Most theoretical models of creative problem-solving suggest a linear progression from preparation and incubation to some type of evaluative study of the "inspiration." These…

[Survey on the medical mycology processes to the members of the French Society for Medical Mycology].

PubMed

Kauffmann-Lacroix, C; Albouy-Llaty, M; Migeot, V; Contet-Audonneau, N

2011-09-01

The objective of the survey was to describe the practices of clinical laboratories in terms of cultures in medical mycology. We have implemented this project within the members of the French Society for Medical Mycology (SFMM) to evaluate the analytical processes of the mycological examination in our laboratories. This preliminary study would help to suggest the future French guidelines. A questionnaire regarding the processing of mycology analysis was sent to the 227 members of the SFMM in 2009. The data involved 21 types of samples, direct microscopic examination with or without colouring and the reagents, the number of culture media, the types of media (Sabouraud, Sabouraud antibiotic, Sabouraud cycloheximide and chromogenic medium), temperature and duration of the incubation (days) and the existence of a first result before the end of the incubation period. The analytical processes were compared to an accredited laboratory according to EN ISO 15189. A great heterogeneity was observed in the 36 forms from 27 (75%) laboratories belonging to university hospitals among the 38 existing in France. As for deep samples, two microscopic exams were performed, only one was usually done. A more sensitive technique was preferred to the wet-mount for some samples. Routine samples are often inoculated on a chromogenic media. For deep samples two medium are inoculated (chromogenic media, Sabouraud and antibiotics). If the temperature of incubation is unique, 30°C was chosen. A temperature of 37°C was preferred for samples where Candida spp. is selected. When there are two temperatures of incubation, 27°C and 37°C were preferred. Each biologist can compare his proceedings to the other laboratories and to a laboratory already accredited. The question is to find the best strategies for each medical mycology specimen. They will aid the process of accreditation according to EN ISO 15189, which now applies in all laboratories in Europe. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

Evaluating propagation method performance over time with Bayesian updating: An application to incubator testing

USGS Publications Warehouse

Converse, Sarah J.; Chandler, J. N.; Olsen, Glenn H.; Shafer, C. C.; Hartup, Barry K.; Urbanek, Richard P.

2010-01-01

In captive-rearing programs, small sample sizes can limit the quality of information on performance of propagation methods. Bayesian updating can be used to increase information on method performance over time. We demonstrate an application to incubator testing at USGS Patuxent Wildlife Research Center. A new type of incubator was purchased for use in the whooping crane (Grus americana) propagation program, which produces birds for release. We tested the new incubator for reliability, using sandhill crane (Grus canadensis) eggs as surrogates. We determined that the new incubator should result in hatching rates no more than 5% lower than the available incubators, with 95% confidence, before it would be used to incubate whooping crane eggs. In 2007, 5 healthy chicks hatched from 12 eggs in the new incubator, and 2 hatched from 5 in an available incubator, for a median posterior difference of <1%, but with a large 95% credible interval (-41%, 43%). In 2008, we implemented a double-blind evaluation method, where a veterinarian determined whether eggs produced chicks that, at hatching, had no apparent health problems that would impede future release. We used the 2007 estimates as priors in the 2008 analysis. In 2008, 7 normal chicks hatched from 15 eggs in the new incubator, and 11 hatched from 15 in an available incubator, for a median posterior difference of 19%, with 95% credible interval (-8%, 44%). The increased sample size has increased our understanding of incubator performance. While additional data will be collected, at this time the new incubator does not appear adequate for use with whooping crane eggs.

Ethanol production from sweet sorghum bagasse through process optimization using response surface methodology.

PubMed

Lavudi, Saida; Oberoi, Harinder Singh; Mangamoori, Lakshmi Narasu

2017-08-01

In this study, comparative evaluation of acid- and alkali pretreatment of sweet sorghum bagasse (SSB) was carried out for sugar production after enzymatic hydrolysis. Results indicated that enzymatic hydrolysis of alkali-pretreated SSB resulted in higher production of glucose, xylose and arabinose, compared to the other alkali concentrations and also acid-pretreated biomass. Response Surface Methodology (RSM) was, therefore, used to optimize parameters, such as alkali concentration, temperature and time of pretreatment prior to enzymatic hydrolysis to maximize the production of sugars. The independent variables used during RSM included alkali concentration (1.5-4%), pretreatment temperature (125-140 °C) and pretreatment time (10-30 min) were investigated. Process optimization resulted in glucose and xylose concentration of 57.24 and 10.14 g/L, respectively. Subsequently, second stage optimization was conducted using RSM for optimizing parameters for enzymatic hydrolysis, which included substrate concentration (10-15%), incubation time (24-60 h), incubation temperature (40-60 °C) and Celluclast concentration (10-20 IU/g-dwt). Substrate concentration 15%, (w/v) temperature of 60 °C, Celluclast concentration of 20 IU/g-dwt and incubation time of 58 h led to a glucose concentration of 68.58 g/l. Finally, simultaneous saccharification fermentation (SSF) as well as separated hydrolysis and fermentation (SHF) was evaluated using Pichia kudriavzevii HOP-1 for production of ethanol. Significant difference in ethanol concentration was not found using either SSF or SHF; however, ethanol productivity was higher in case of SSF, compared to SHF. This study has established a platform for conducting scale-up studies using the optimized process parameters.

Comparison Study of Different BOD Tests in the Determination of BOD7 Evaluated in a Model Domestic Sewage

PubMed Central

Roppola, Katri; Kuokkanen, Toivo; Rämö, Jaakko; Prokkola, Hanna; Heiska, Eeva

2007-01-01

The aim of this research was to study the suitability of a manometric respirometric test (OECD 301F) for the determination of biological oxygen demand (BOD) in domestic wastewater. This was done by comparing four different BOD tests (a manometric respirometric test, oxygen sensor determination, iodometric titration, and UV absorption measurement) and by evaluating the BOD7 value after a short-term measurement applying the manometric respirometric test. Measurements were done in three different laboratories. The respirometric test was performed well on municipal sewage, and the precision of results was good. There were minor deviations between BOD7 values as determined by the different methods, but the deviations were not essentially big enough to have a practical impact. Evaluation of the BOD7 value after two or three days' incubation by the respirometric method was shown to work well; average deviations between measured and evaluated values after three days' incubation were 5–18%. Hence, the manometric respirometric test proved to be a rapid and suitable tool for process control purposes. PMID:17671608

Incubation: A Neglected Aspect of the Writing Process.

ERIC Educational Resources Information Center

Krashen, Stephen

2001-01-01

Discusses the role of incubation in the writing process. Suggests that one secret to coming up with good ideas in writing is understanding the importance of realizing that the process entails patient revision, takes time, and often requires some time off task. (Author/VWL)

A weak electric field-assisted ultrafast electrical switching dynamics in In3SbTe2 phase-change memory devices

NASA Astrophysics Data System (ADS)

Pandey, Shivendra Kumar; Manivannan, Anbarasu

2017-07-01

Prefixing a weak electric field (incubation) might enhance the crystallization speed via pre-structural ordering and thereby achieving faster programming of phase change memory (PCM) devices. We employed a weak electric field, equivalent to a constant small voltage (that is incubation voltage, Vi of 0.3 V) to the applied voltage pulse, VA (main pulse) for a systematic understanding of voltage-dependent rapid threshold switching characteristics and crystallization (set) process of In3SbTe2 (IST) PCM devices. Our experimental results on incubation-assisted switching elucidate strikingly one order faster threshold switching, with an extremely small delay time, td of 300 ps, as compared with no incubation voltage (Vi = 0 V) for the same VA. Also, the voltage dependent characteristics of incubation-assisted switching dynamics confirm that the initiation of threshold switching occurs at a lower voltage of 0.82 times of VA. Furthermore, we demonstrate an incubation assisted ultrafast set process of IST device for a low VA of 1.7 V (˜18 % lesser compared to without incubation) within a short pulse-width of 1.5 ns (full width half maximum, FWHM). These findings of ultrafast switching, yet low power set process would immensely be helpful towards designing high speed PCM devices with low power operation.

High- and low-temperature manipulation during late incubation: effects on embryonic development, the hatching process, and metabolism in broilers.

PubMed

Willemsen, H; Kamers, B; Dahlke, F; Han, H; Song, Z; Ansari Pirsaraei, Z; Tona, K; Decuypere, E; Everaert, N

2010-12-01

Temperatures continuously higher and lower than the standard incubation temperature by 3°C from embryonic d 16 until embryonic d 18.5 result in differential effects on embryonic development, the hatching process, and embryonic metabolism. Embryos in the high-temperature group were forced into a state of malnutrition by the temperature treatment, as reflected by reduced embryo growth and yolk consumption, resulting in a significantly lower chick weight at hatch. In addition, altered air cell and blood gases as well as a retarded hatching process further indicated reduced growth of embryos exposed to higher incubation temperatures during the latter part of incubation. In addition, hatchability was significantly reduced by the high-temperature treatment due to higher embryonic mortality during the treatment period and the hatching process. Levels of blood glucose, lactate, liver glycogen, plasma triglycerides, and nonesterified fatty acids indicated an altered carbohydrate and lipid metabolism for the high-temperature group. Although the hatching process of embryos exposed to lower incubation temperatures was also significantly retarded, their embryonic development and growth were strikingly similar to those of the control group.

A review of factors influencing the availability of dissolved oxygen to incubating salmonid embryos

NASA Astrophysics Data System (ADS)

Greig, S. M.; Sear, D. A.; Carling, P. A.

2007-01-01

Previous investigations into factors influencing incubation success of salmonid progeny have largely been limited to the development of empirical relationships between characteristics of the incubation environment and survival to emergence. It is suggested that adopting a process-based approach to assessing incubation success aids identification of the precise causes of embryonic mortalities, and provides a robust framework for developing and implementing managerial responses.Identifying oxygen availability within the incubation environment as a limiting factor, a comprehensive review of trends in embryonic respiration, and processes influencing the flux of oxygenated water through gravel riverbeds is provided. The availability of oxygen to incubating salmonid embryos is dependent on the exchange of oxygenated water with the riverbed, and the ability of the riverbed gravel medium to transport this water at a rate and concentration appropriate to support embryonic respiratory requirements. Embryonic respiratory trends indicate that oxygen consumption varies with stage of development, ambient water temperature and oxygen availability. The flux of oxygenated water through the incubation environment is controlled by a complex interaction of intragravel and extragravel processes and factors. The processes driving the exchange of channel water with gravel riverbeds include bed topography, bed permeability, and surface roughness effects. The flux of oxygenated water through riverbed gravels is controlled by gravel permeability, coupling of surface-subsurface flow and oxygen demands imposed by materials infiltrating riverbed gravels. Temporally and spatially variable inputs of groundwater can also influence the oxygen concentration of interstitial water. Copyright

Evaluation of microbial community composition in thermophilic methane-producing incubation of production water from a high-temperature oil reservoir.

PubMed

Zhou, Fang; Mbadinga, Serge Maurice; Liu, Jin-Feng; Gu, Ji-Dong; Mu, Bo-Zhong

2013-01-01

Investigation of petroleum microbes is fundamental for the development and utilization of oil reservoirs' microbial resources, and also provides great opportunities for research and development of bio-energy. Production water from a high-temperature oil reservoir was incubated anaerobically at 55 degrees C for more than 400 days without amendment of any nutrients. Over the time of incubation, about 1.6 mmol of methane and up to 107 micromol of hydrogen (H2) were detected in the headspace. Methane formation indicated that methanogenesis was likely the predominant process in spite of the presence of 23.4 mM SO4(2-) in the production water. Microbial community composition of the incubation was characterized by means of 16S rRNA gene clone libraries construction. Bacterial composition changed from Pseudomonales as the dominant population initially to Hydrogenophilales-related microorganisms affiliated to Petrobacter spp. closely. After 400 days of incubation, other bacterial members detected were related to Anareolineales, beta-, gamma-, and delta-Proteobacteria. The archaeal composition of the original production water was essentially composed of obligate acetoclastic methanogens of the genus Methanosaeta, but the incubation was predominantly composed of CO2-reducing methanogens of the genus Methanothermobacter and Crenarchaeotes-related microorganisms. Our results suggest that methanogenesis could be more active than expected in oil reservoir environments and methane formation from CO2-reduction played a significant role in the methanogenic community. This conclusion is consistent with the predominant role played by H2-oxidizing methanogens in the methanogenic conversion of organic matter in high-temperature petroleum reservoirs.

Incubation temperature and time of hatch impact broiler muscle growth and morphology.

PubMed

Clark, D L; Walter, K G; Velleman, S G

2017-09-01

The adult myogenic population of stem cells, called satellite cells, initially develop in late-term embryos. Satellite cells are the only myogenic cell that repair damaged myofibers and increase post-hatch growth. The objective of the current study was to determine if incubation temperatures and time of hatch impact growth and pectoralis major (p. major) muscle morphology. Eggs were incubated at a constant 37.8°C; however, from d 14 to 18, the eggs were subject to 39.5°C for 0, 3, or 12 h per day. Chicks were divided into early, mid, or late hatch groups based upon the time they emerged from the shell. Growth and feed efficiency were measured throughout the 63-day trial, while meat quality and muscle morphology were evaluated at the time of processing. The chicks incubated at an increased temperature for 12 h per d had reduced (P < 0.01) body weights throughout the trial compared to the 3 h treatment and control. The early hatch broilers were heavier (P < 0.01) at 63 d compared to mid and late hatch broilers. Chicks from the 12 h incubation treatment had an increased (P = 0.01) gain to feed ratio compared to the control. Broilers from the 12 h incubation treatment had lower (P < 0.01) p. major weights compared to the 0 and 3 h treatments. Early hatch broilers had heavier p. major weights (P < 0.01) compared to mid and late hatch groups. The 12 h incubation treatment also reduced the number of broilers with moderate to severe myopathic attributes compared to the control. Similarly, there were fewer late hatch birds with fibrotic and necrotic p. major muscles compared to the early hatch group. Together, these data demonstrate that altering incubation temperature is a feasible management strategy to improve muscle morphology without negatively impacting meat quality parameters. © 2017 Poultry Science Association Inc.

Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers.

PubMed

Carretero, M I; Fumuso, F G; Neild, D M; Giuliano, S M; Cetica, P; Miragaya, M H

2015-09-01

The objectives of this study were to evaluate the effect of different acrosome reaction (AR) inducers on viability and acrosomal status in llama spermatozoa, by using the FITC-PNA/PI technique and evaluate if there is a positive correlation between the FITC-PNA/PI and the Coomassie blue (CB) staining techniques. After incubating twenty ejaculates in 0.1% collagenase the centrifuged pellets were resuspended in TALP-BSA medium. An aliquot was sonicated to remove the acrosomal content (positive control). The rest of the sample was incubated for 3h at 38 °C with 5% CO2 and 100% humidity. Three aliquots were further incubated 1h with one of the following AR inducers: calcium ionophore, ionomycin or progesterone. One without inducers and the other, incubated with dimethyl sulfoxide (vehicle of the inducing agents). Acrosomes were evaluated at time 0 and after 4h incubation. Calcium ionophore was the most potent agent for inducing the AR (67.2 ± 14.4% live+dead AR sperm) (P < 0.05). These samples showed no motility and viability was very low (0-30%). Both ionomycin and progesterone presented significantly higher (P < 0.05) percentages of total AR sperm than the controls, but had similar percentages of dead reacted sperm to the controls. A positive correlation was observed between the intact acrosome FITC-PNA/PI pattern (live+dead sperm) and the acrosome-present CB pattern (r = 0.64; P = 0.000) in all the evaluated samples. the FITC-PNA/PI technique simultaneously evaluates viability and acrosomal status in llama spermatozoa and calcium ionophore could be used as a control of AR. Copyright © 2015 Elsevier B.V. All rights reserved.

Use of agar diffusion assay to evaluate bactericidal activity of formulations of alkaline salts of fatty acids against bacteria associated with poultry processing

USDA-ARS?s Scientific Manuscript database

The agar diffusion assay was used to examine antibacterial activity of alkaline salts of fatty acids (FA). Wells in agar media seeded with bacteria were filled with FA-potassium hydroxide (KOH) solutions, plates were incubated, and zones of inhibition were measured. The relationship between bacteric...

Organic Matter Loading Modifies the Microbial Community Responsible for Nitrogen Loss in Estuarine Sediments.

PubMed

Babbin, Andrew R; Jayakumar, Amal; Ward, Bess B

2016-04-01

Coastal marine sediments, as locations of substantial fixed nitrogen loss, are very important to the nitrogen budget and to the primary productivity of the oceans. Coastal sediment systems are also highly dynamic and subject to periodic natural and anthropogenic organic substrate additions. The response to organic matter by the microbial community involved in nitrogen loss processes was evaluated using mesocosms of Chesapeake Bay sediments. Over the course of a 50-day incubation, rates of anammox and denitrification were measured weekly using (15)N tracer incubations, and samples were collected for genetic analysis. Rates of both nitrogen loss processes and gene abundances associated with them corresponded loosely, probably because heterogeneities in sediments obscured a clear relationship. The rates of denitrification were stimulated more, and the fraction of nitrogen loss attributed to anammox slightly reduced, by the higher organic matter addition. Furthermore, the large organic matter pulse drove a significant and rapid shift in the denitrifier community composition as determined using a nirS microarray, indicating that the diversity of these organisms plays an essential role in responding to anthropogenic inputs. We also suggest that the proportion of nitrogen loss due to anammox in these coastal estuarine sediments may be underestimated due to temporal dynamics as well as from methodological artifacts related to conventional sediment slurry incubation approaches.

Low pH inactivation for xenotropic gamma retrovirus in recombinant human TNF-α receptor immunoglobulin G and mechanism of inactivation.

PubMed

Ma, Rong; Cui, Xiaolan

2014-01-01

CHO-derived recombinant proteins for human therapeutic are used commonly. There are noninfectious endogenous retroviruses in CHO cells. Validation study for inactivation process is required. Murine xenotropic gamma retrovirus (X-MulV) is a model virus in validation study. In our previous study, optimum conditions for X-MulV inactivation were sifted. In this study, we performed a further research on low pH inactivation for evaluation of X-MulV clearance in manufacturing of recombinant human TNF-α receptor immunoglobulin G fusion proteins (rhTNF-α) for injection. Cell-based infectivity assay was used for the evaluation of X-MulV clearance. RhTNF-α were spiked with X-MulV and were inactivated at pH 3.60 ∼ 3.90, 25 ± 2 °C, and 0 ∼ 240 min, respectively. Samples incubated at the conditions for 15 ∼ 180 min were not inactivated effectively. For 4 h incubation, log10 reductions were achieved 5.0 log10. Biological activity of rhTNF-α incubated at pH 3.60, 25 °C for 4 h, which was assayed on murine L929 fibroblasts cells, was not affected by low pH. Env gene of X-MulV, which was detected by conventional PCR method for the first time, was not detected after incubation at pH 3.60, and it may be the mechanism of low pH inactivation. Copyright © 2013. Published by Elsevier Ltd.

Evaluation of incubation time for dermatophytes cultures.

PubMed

Rezusta, Antonio; de la Fuente, Sonia; Gilaberte, Yolanda; Vidal-García, Matxalen; Alcalá, Leticia; López-Calleja, Ana; Ruiz, Maria Angeles; Revillo, Maria José

2016-07-01

In general, it is recommended to incubate dermatophytes cultures for a minimum of 4 weeks. Several aspects of routine fungal cultures should be evaluated in order to implement appropriate and necessary changes. The aim of this study was to determine the optimum incubation time for routine dermatophytes cultures, analysing the time to find first fungal growth by visual observation. We recorded the time when the initial growth was detected for all dermatophyte isolates during a 4-year period. A total of 5459 dermatophyte cultures were submitted to our laboratory. From the total cultures, only 16 (1.42%) isolates were recovered over/after 17 days of incubation and only three dermatophyte species were recovered over 17 days. Fourteen isolates belong to Trichophyton rubrum, one isolate to Trichophyton mentagrophytes complex and one isolate to Epidermophyton floccosum. We concluded that an incubation period of 17 days is enough to establish a microbiological diagnosis of dermatophytosis. © 2016 Blackwell Verlag GmbH.

Measuring efficiency and productivity growth of new technology-based firms in business incubators: the Portuguese case study of Madan Parque.

PubMed

Grilo, A; Santos, J

2015-01-01

Business incubators can play a major role in helping to turn a business idea into a technology-based organization that is economically efficient. However, there is a shortage in the literature regarding the efficiency evaluation and productivity evolution of the new technology-based firms (NTBFs) in the incubation scope. This study develops a model based on the data envelopment analysis (DEA) methodology, which allows the incubated NTBFs to evaluate and improve the efficiency of their management. Moreover, the Malmquist index is used to examine productivity change. The index is decomposed into multiple components to give insights into the root sources of productivity change. The proposed model was applied in a case study with 13 NTBFs incubated. From that study, we conclude that inefficient firms invest excessively in research and development (R&D), and, on average, firms have a productivity growth in the period of study.

Measuring Efficiency and Productivity Growth of New Technology-Based Firms in Business Incubators: The Portuguese Case Study of Madan Parque

PubMed Central

Grilo, A.; Santos, J.

2015-01-01

Business incubators can play a major role in helping to turn a business idea into a technology-based organization that is economically efficient. However, there is a shortage in the literature regarding the efficiency evaluation and productivity evolution of the new technology-based firms (NTBFs) in the incubation scope. This study develops a model based on the data envelopment analysis (DEA) methodology, which allows the incubated NTBFs to evaluate and improve the efficiency of their management. Moreover, the Malmquist index is used to examine productivity change. The index is decomposed into multiple components to give insights into the root sources of productivity change. The proposed model was applied in a case study with 13 NTBFs incubated. From that study, we conclude that inefficient firms invest excessively in research and development (R&D), and, on average, firms have a productivity growth in the period of study. PMID:25874266

Simulation of prenatal maternal sounds in NICU incubators: a pilot safety and feasibility study.

PubMed

Panagiotidis, John; Lahav, Amir

2010-10-01

This pilot study evaluated the safety and feasibility of an innovative audio system for transmitting maternal sounds to NICU incubators. A sample of biological sounds, consisting of voice and heartbeat, were recorded from a mother of a premature infant admitted to our unit. The maternal sounds were then played back inside an unoccupied incubator via a specialized audio system originated and compiled in our lab. We performed a series of evaluations to determine the safety and feasibility of using this system in NICU incubators. The proposed audio system was found to be safe and feasible, meeting criteria for humidity and temperature resistance, as well as for safe noise levels. Simulation of maternal sounds using this system seems achievable and applicable and received local support from medical staff. Further research and technology developments are needed to optimize the design of the NICU incubators to preserve the acoustic environment of the womb.

Effect of agitation and terminal subcultures on yield and speed of detection of the Oxoid Signal blood culture system versus the BACTEC radiometric system

DOE Office of Scientific and Technical Information (OSTI.GOV)

Weinstein, M.P.; Mirrett, S.; Reimer, L.G.

1989-03-01

In an initial evaluation, we found the Oxoid Signal blood culture system inferior to the BACTEC radiometric system for detection of some microorganisms causing septicemia. To determine whether modified processing of the Oxoid Signal blood culture system could improve its yield and speed of detecting positive cultures relative to the BACTEC radiometric system, we agitated all Oxoid bottles during the first 24 to 48 h of incubation and performed aerobic and anaerobic subcultures of all Oxoid bottles negative after 7 days of incubation. These modifications improved the overall performance of the Oxoid system, particularly with regard to the yield ofmore » streptococci, members of the family Enterobacteriaceae, and Haemophilus, Neisseria, and Acinetobacter spp. The speed of detecting positive cultures also was improved, especially within the first 24 h of incubation. However, the BACTEC system still detected more positive cultures (P less than 0.005), especially of obligate aerobes such as Pseudomonas aeruginosa (P less than 0.05) and yeasts (P less than 0.005). The BACTEC system also detected positive cultures earlier than the Oxoid system (e.g., at 24 h of incubation, 70.5% of BACTEC positive cultures detected versus 62.1% of Oxoid positive cultures detected). Further modifications of the Oxoid system which might include a revised medium, additional processing modifications, altered headspace atmosphere, or a complementary second broth medium should be considered, since the system is attractive in concept and is easy to use in the clinical laboratory.« less

Advanced Oxidation Process sanitization of hatching eggs reduces Salmonella in broiler chicks.

PubMed

Rehkopf, A C; Byrd, J A; Coufal, C D; Duong, T

2017-10-01

The microbial quality of eggs entering the hatchery is an important critical control point for biosecurity, pathogen reduction, and food safety programs in poultry production. Developing interventions to reduce Salmonella contamination of eggs is important to improving the microbial food safety of poultry and poultry products. The hydrogen peroxide (H2O2) and ultraviolet light (UV) Advanced Oxidation Process (AOP) has been previously demonstrated to be effective in reducing Salmonella on the surface of experimentally contaminated eggs. The objective of this study was to evaluate the effect of treating eggs with an egg-sanitizing apparatus using the H2O2/UV AOP on Salmonella contamination during incubation, hatching, and in broiler chicks during grow-out. Experimentally contaminated eggs were treated using the automated H2O2/UV AOP egg sanitizer and incubated for 21 d. AOP sanitization reduced Salmonella up to 7 log10 cfu egg-1 (P < 0.05) from the surface of experimentally contaminated eggs and reduced the number of Salmonella positive eggs by up to 75% (P < 0.05) when treated 1 h post-inoculation. AOP treatment also reduced the number of Salmonella-positive eggs during incubation. Additionally, Salmonella was recovered from more chicks hatched from untreated eggs than from eggs treated using the H2O2/UV AOP egg sanitizer (P < 0.05) through 14 d posthatch. These data suggest reduction of Salmonella contamination on the surface of eggs using the H2O2/UV AOP egg sanitizer prior to incubation may reduce the gastrointestinal colonization of chicks by Salmonella. © 2017 Poultry Science Association Inc.

Waste vinegar residue as substrate for phytase production.

PubMed

Wang, Zhi-Hong; Dong, Xiao-Fang; Zhang, Guo-Qing; Tong, Jian-Ming; Zhang, Qi; Xu, Shang-Zhong

2011-12-01

Waste vinegar residue, the by-product of vinegar processing, was used as substrate for phytase production from Aspergillus ficuum NTG-23 in solid-state fermentation to investigate the potential for the efficient re-utilization or recycling of waste vinegar residue. Statistical designs were applied in the processing of phytase production. First, a Plackett-Burman (PB) design was used to evaluate eleven parameters: glucose, starch, wheat bran, (NH(4))(2)SO(4), NH(4)NO(3), tryptone, soybean meal, MgSO(4)·7H(2)O, CaCl(2)·7H(2)O, FeSO(4)·7H(2)O, incubation time. The PB experiments showed that there were three significant factors: glucose, soybean meal and incubation time. The closest values to the optimum point were then derived by steepest ascent path. Finally, a mathematical model was created and validated to explain the behavioural process after these three significant factors were optimized using response surface methodology (RSM). The best phytase activity was attained using the following conditions: glucose (7.2%), soybean meal (5.1%), and incubation time (271 h). The phytase activity was 7.34-fold higher due to optimization by PB design, steepest ascent path design and RSM. The phytase activity was enhanced 0.26-fold in comparison with the results by the second step of steepest ascent path design. The results indicate that with waste vinegar residue as a substrate higher production of phytase from Aspergillus ficuum NTG-23 could be obtained through an optimization process and that this method might be applied to an integrated system for recycling of the waste vinegar residue.

Utilizing pretreatment and fungal incubation to enhance the nutritional value of canola meal.

PubMed

Croat, J R; Karki, B; Berhow, M; Iten, L; Muthukumarappan, K; Gibbons, W R

2017-08-01

The objective of this study was to determine the optimal pretreatment and fungal strain to reduce glucosinolates (GLS), fibre and residual sugars while increasing the nutritional value of canola meal. Submerged incubation conditions were used to evaluate four pretreatment methods (extrusion, hot water cook, dilute acid and dilute alkali) and three fungal cultures (Aureobasidium pullulans Y-2311-1, Fusarium venenatum NRRL-26139 and Trichoderma reesei NRRL-3653) in hexane-extracted (HE) and cold-pressed (CP) canola meal. The combination of extrusion pretreatment followed by incubation with T. reesei resulted in the greatest overall improvement to HE canola meal, increasing protein to 51·5%, while reducing NDF, GLS and residual sugars to 18·6%, 17·2 μmol l -1  g -1 and 5% w/w, respectively. Extrusion pretreatment and incubation with F. venenatum performed the best with CP canola meal, resulting in 54·4% protein while reducing NDF, GLS and residual sugars to 11·6%, 6·7 μmol l -1  g -1 and 3·8% w/w respectively. The work is significant in that it provides a method of reducing GLS (up to 98%) and neutral detergent fibre (up to 65%) while increasing the protein content (up to 45%) of canola meal. This novel pretreatment and submerged incubation process could be used to produce a canola product with higher nutritional value for livestock consumption. © 2017 The Society for Applied Microbiology.

GENESA as an Aid to Incubation/Imagery.

ERIC Educational Resources Information Center

Bruch, Catherine; And Others

1979-01-01

The article focuses on the applications of the GENESA model (a life sized model of the geometry of a biological cell) in the enhancement of the creative processes during the stages of incubation, illumination, and verification, with emphasis primarily on the phase of incubation/imagery through potential illumination. (SBH)

Extremely low frequency (ELF) stray magnetic fields of laboratory equipment: a possible co-exposure conducting experiments on cell cultures.

PubMed

Gresits, Iván; Necz, Péter Pál; Jánossy, Gábor; Thuróczy, György

2015-09-01

Measurements of extremely low frequency (ELF) magnetic fields were conducted in the environment of commercial laboratory equipment in order to evaluate the possible co-exposure during the experimental processes on cell cultures. Three types of device were evaluated: a cell culture CO2 incubator, a thermostatic water bath and a laboratory shaker table. These devices usually have electric motors, heating wires and electronic control systems, therefore may expose the cell cultures to undesirable ELF stray magnetic fields. Spatial distributions of magnetic field time domain signal waveform and frequency spectral analysis (FFT) were processed. Long- and short-term variation of stray magnetic field was also evaluated under normal use of investigated laboratory devices. The results show that the equipment under test may add a considerable ELF magnetic field to the ambient environmental magnetic field or to the intentional exposure to ELF, RF or other physical/chemical agents. The maximum stray magnetic fields were higher than 3 µT, 20 µT and 75 µT in the CO2 incubator, in water bath and on the laboratory shaker table, respectively, with high variation of spatial distribution and time domain. Our investigation emphasizes possible confounding factors conducting cell culture studies related to low-level ELF-EMF exposure due to the existing stray magnetic fields in the ambient environment of laboratory equipment.

Antioxidant and anti-inflammatory activity of a flavonoid-rich concentrate recovered from Opuntia ficus-indica juice.

PubMed

Matias, A; Nunes, S L; Poejo, J; Mecha, E; Serra, A T; Madeira, Paulo J Amorim; Bronze, M R; Duarte, C M M

2014-12-01

In this work, Opuntia ficus indica juice was explored as a potential source of natural antioxidant and anti-inflammatory ingredients towards intestinal inflammation. An adsorption separation process was used to produce a natural flavonoid-rich concentrate (FRC) from Opuntia ficus-indica juice. The FRC effect (co- or pre-incubation) on induced-oxidative stress and induced-inflammation was evaluated in human Caco-2 cells. The main constituents identified and present in the extract are flavonoids (namely isorhamnetins and their derivatives such as isorhamnetin 3-O-rhamnose-rutinoside and isorhamnetin 3-O-rutinoside) and phenolic acids (such as ferulic, piscidic and eucomic acids). Our results showed that co-incubation of FRC with the stress-inducer attenuates radicals production in a much more significant manner than pre-incubation. These results suggest that FRC compounds which cannot pass the cell membrane freely (isorhamnetin derivatives) have an ability to inhibit the formation of H2O2-induced radicals in the surrounding environment of intestinal epithelial cells. The capacity of FRC (co-incubation) for suppressing (at the extracellular level) free radicals chain initiation or propagation reaction was probably related with a more pronounced reduction in protein oxidation. A similar response was observed in the inflammatory state, where a marked decrease in IL-8 secretion and blocked degradation of IκBα was achieved for FRC co-incubation. Simultaneously, treatment with FRC significantly reduces NO and TNF-α expression and modulates apparent permeability in Caco-2 cells. In these cases, no significant differences were found between pre- and co-incubation treatments suggesting that bioavailable phenolics, such as ferulic, eucomic and piscidic acids and isorhamnetin, act at the intracellular environment.

Learning Incubator: an instrument to foster entrepreneurship in Nursing.

PubMed

Backes, Dirce Stein; Obem, Marielle Kulakowski; Pereira, Simone Barbosa; Gomes, Carine Alves; Backes, Marli Terezinha Stein; Erdmann, Alacoque Lorenzini

2015-01-01

this study aimed to know the contributions of the Learning Incubator to the process of lifelong education in health. this is a qualitative field research whose data was collected from August to December 2014 by the focus group technique. The research had 34 employees of a Teaching Hospital in the central region of the state of Rio Grande do Sul that participated previously in the incubation process. from the data encoded by content analysis, three themes were selected: Learning Incubator - welcoming and integrating space; An instigating instrument that enhances possibilities; Continuous and lifelong education strategy. the Learning Incubator is an important instrument to foster entrepreneurship in nursing and other health areas due to its capacity of rethinking mechanized practices, to the possibility of instigating new ways of being and acting, and to the ability of creating and developing new ideas based on individual and institutional needs.

Development of the e-Baby serious game with regard to the evaluation of oxygenation in preterm babies: contributions of the emotional design.

PubMed

Fonseca, Luciana Mara Monti; Dias, Danielle Monteiro Vilela; Góes, Fernanda Dos Santos Nogueira; Seixas, Carlos Alberto; Scochi, Carmen Gracinda Silvan; Martins, José Carlos Amado; Rodrigues, Manuel Alves

2014-09-01

The present study aimed to describe the development process of a serious game that enables users to evaluate the respiratory process in a preterm infant based on an emotional design model. The e-Baby serious game was built to feature the simulated environment of an incubator, in which the user performs a clinical evaluation of the respiratory process in a virtual preterm infant. The user learns about the preterm baby's history, chooses the tools for the clinical evaluation, evaluates the baby, and determines whether his/her evaluation is appropriate. The e-Baby game presents phases that contain respiratory process impairments of higher or lower complexity in the virtual preterm baby. Included links give the user the option of recording the entire evaluation procedure and sharing his/her performance on a social network. e-Baby integrates a Clinical Evaluation of the Preterm Baby course in the Moodle virtual environment. This game, which evaluates the respiratory process in preterm infants, could support a more flexible, attractive, and interactive teaching and learning process that includes simulations with features very similar to neonatal unit realities, thus allowing more appropriate training for clinical oxygenation evaluations in at-risk preterm infants. e-Baby allows advanced user-technology-educational interactions because it requires active participation in the process and is emotionally integrated.

Evaluation of the Biolog MicroStation system for yeast identification

NASA Technical Reports Server (NTRS)

McGinnis, M. R.; Molina, T. C.; Pierson, D. L.; Mishra, S. K.

1996-01-01

One hundred and fifty-nine isolates representing 16 genera and 53 species of yeasts were processed with the Biolog MicroStation System for yeast identification. Thirteen genera and 38 species were included in the Biolog database. For these 129 isolates, correct identifications to the species level were 13.2, 39.5 and 48.8% after 24, 48 and 72 hours incubation at 30 degrees C, respectively. Three genera and 15 species which were not included in the Biolog database were also tested. Of the 30 isolates studied, 16.7, 53.3 and 56.7% of the isolates were given incorrect names from the system's database after 24,48 and 72 h incubation at 30 degrees C, respectively. The remaining isolates of this group were not identified.

A Handy Preterm Infant Incubator for Providing Intensive Care: Simulation, 3D Printed Prototype, and Evaluation.

PubMed

Zaylaa, Amira J; Rashid, Mohamad; Shaib, Mounir; El Majzoub, Imad

2018-01-01

Preterm infants encounter an abrupt delivery before their complete maturity during the third trimester of pregnancy. Polls anticipate an increase in the rates of preterm infants for 2025, especially in middle- and low-income countries. Despite the abundance of intensive care methods for preterm infants, such as, but not limited to, commercial, transport, embrace warmer, radiant warmer, and Kangaroo Mother Care methods, they are either expensive, lack the most essential requirements or specifications, or lack the maternal-preterm bond. This drove us to carry this original research and innovative idea of developing a new 3D printed prototype of a Handy preterm infant incubator. We aim to provide the most indispensable intensive care with the lowest cost, to bestow low-income countries with the Handy incubator's care, preserve the maternal -preterm's bond, and diminish the rate of mortality. Biomedical features, electronics, and biocompatible materials were utilized. The design was simulated, the prototype was 3D printed, and the outcomes were tested and evaluated. Simulation results showed the best fit for the Handy incubator's components. Experimental results showed the 3D-printed prototype and the time elapsed to obtain it. Evaluation results revealed that the overall performance of Kangaroo Mother Care and the embrace warmer was 75 ± 1.4% and 66.7 ± 1.5%, respectively, while the overall performance of our Handy incubator was 91.7 ± 1.6%, thereby our cost-effective Handy incubator surpassed existing intensive care methods. The future step is associating the Handy incubator with more specifications and advancements.

Oligomers, protofibrils and amyloid fibrils from recombinant human lysozyme (rHL): fibrillation process and cytotoxicity evaluation for ARPE-19 cell line.

PubMed

Ruiz, Eva D; Almada, Mario; Burboa, María G; Taboada, Pablo; Mosquera, Víctor; Valdez, Miguel A; Juárez, Josué

2015-02-01

Amyloid-associated diseases, such Alzheimer's, Huntington's, Parkinson's, and type II diabetes, are related to protein misfolding and aggregation. Herein, the time evolution of scattered light intensity, hydrophobic properties, and conformational changes during fibrillation processes of rHL solutions at 55 °C and pH 2.0 were used to monitor the aggregation process of recombinant human lysozyme (rHL). Dynamic light scattering (DLS), thioflavin T (ThT) fluorescence, and surface tension (ST) at the air-water interface were used to analyze the hydrophobic properties of pre-amyloid aggregates involved in the fibrillation process of rHL to find a correlation between the hydrophobic character of oligomers, protofibrils and amyloid aggregates with the gain in cross-β-sheet structure, depending on the increase in the incubation periods. The ability of the different aggregates of rHL isolated during the fibrillation process to be adsorbed at the air-water interface can provide important information about the hydrophobic properties of the protein, which can be related to changes in the secondary structure of rHL, resulting in cytotoxic or non-cytotoxic species. Thus, we evaluated the cytotoxic effect of oligomers, protofibrils and amyloid fibrils on the cell line ARPE-19 using the MTT reduction test. The more cytotoxic protein species arose after a 600-min incubation time, suggesting that the hydrophobic character of pre-amyloid fibrils, in addition to the high prevalence of the cross-β-sheet conformation, can become toxic for the cell line ARPE-19. Copyright © 2014 Elsevier B.V. All rights reserved.

An evaluation of three processing methods and the effect of reduced culture times for faster direct identification of pathogens from BacT/ALERT blood cultures by MALDI-TOF MS.

PubMed

Loonen, A J M; Jansz, A R; Stalpers, J; Wolffs, P F G; van den Brule, A J C

2012-07-01

Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) is a fast and reliable method for the identification of bacteria from agar media. Direct identification from positive blood cultures should decrease the time to obtaining the result. In this study, three different processing methods for the rapid direct identification of bacteria from positive blood culture bottles were compared. In total, 101 positive aerobe BacT/ALERT bottles were included in this study. Aliquots from all bottles were used for three bacterial processing methods, i.e. the commercially available Bruker's MALDI Sepsityper kit, the commercially available Molzym's MolYsis Basic5 kit and a centrifugation/washing method. In addition, the best method was used to evaluate the possibility of MALDI application after a reduced incubation time of 7 h of Staphylococcus aureus- and Escherichia coli-spiked (1,000, 100 and 10 colony-forming units [CFU]) aerobe BacT/ALERT blood cultures. Sixty-six (65%), 51 (50.5%) and 79 (78%) bottles were identified correctly at the species level when the centrifugation/washing method, MolYsis Basic 5 and Sepsityper were used, respectively. Incorrect identification was obtained in 35 (35%), 50 (49.5%) and 22 (22%) bottles, respectively. Gram-positive cocci were correctly identified in 33/52 (64%) of the cases. However, Gram-negative rods showed a correct identification in 45/47 (96%) of all bottles when the Sepsityper kit was used. Seven hours of pre-incubation of S. aureus- and E. coli-spiked aerobe BacT/ALERT blood cultures never resulted in reliable identification with MALDI-TOF MS. Sepsityper is superior for the direct identification of microorganisms from aerobe BacT/ALERT bottles. Gram-negative pathogens show better results compared to Gram-positive bacteria. Reduced incubation followed by MALDI-TOF MS did not result in faster reliable identification.

Embryonic developmental patterns and energy expenditure are affected by incubation temperature in wood ducks (Aix sponsa).

PubMed

DuRant, S E; Hopkins, W A; Hepp, G R

2011-01-01

Recent research in birds has demonstrated that incubation temperature influences a suite of traits important for hatchling development and survival. We explored a possible mechanism for the effects on hatchling quality by determining whether incubation temperature influences embryonic energy expenditure of wood ducks (Aix sponsa). Because avian embryos are ectothermic, we hypothesized that eggs incubated at higher temperatures would have greater energy expenditure at any given day of incubation. However, because eggs incubated at lower temperatures take longer to hatch than embryos incubated at higher temperatures, we hypothesized that the former would expend more energy during incubation. We incubated eggs at three temperatures (35.0°, 35.9°, and 37.0°C) that fall within the range of temperatures of naturally incubated wood duck nests. We then measured the respiration of embryos every 3 d during incubation, immediately after ducks externally pipped, and immediately after hatching. As predicted, embryos incubated at the highest temperature had the highest metabolic rates on most days of incubation, and they exhibited faster rates of development. Yet, because of greater energy expended during the hatching process, embryos incubated at the lowest temperature expended 20%-37% more energy during incubation than did embryos incubated at the higher temperatures. Slower developmental rates and greater embryonic energy expenditure of embryos incubated at the lowest temperature could contribute to their poor physiological performance as ducklings compared with ducklings that hatch from eggs incubated at higher temperatures.

Information content of incubation experiments for inverse estimation of pools in the Rothamsted carbon model: a Bayesian approach

NASA Astrophysics Data System (ADS)

Scharnagl, Benedikt; Vrugt, Jasper A.; Vereecken, Harry; Herbst, Michael

2010-05-01

Turnover of soil organic matter is usually described with multi-compartment models. However, a major drawback of these models is that the conceptually defined compartments (or pools) do not necessarily correspond to measurable soil organic carbon (SOC) fractions in real practice. This not only impairs our ability to rigorously evaluate SOC models but also makes it difficult to derive accurate initial states. In this study, we tested the usefulness and applicability of inverse modeling to derive the various carbon pool sizes in the Rothamsted carbon model (ROTHC) using a synthetic time series of mineralization rates from laboratory incubation. To appropriately account for data and model uncertainty we considered a Bayesian approach using the recently developed DiffeRential Evolution Adaptive Metropolis (DREAM) algorithm. This Markov chain Monte Carlo scheme derives the posterior probability density distribution of the initial pool sizes at the start of incubation from observed mineralization rates. We used the Kullback-Leibler divergence to quantify the information contained in the data and to illustrate the effect of increasing incubation times on the reliability of the pool size estimates. Our results show that measured mineralization rates generally provide sufficient information to reliably estimate the sizes of all active pools in the ROTHC model. However, with about 900 days of incubation, these experiments are excessively long. The use of prior information on microbial biomass provided a way forward to significantly reduce uncertainty and required duration of incubation to about 600 days. Explicit consideration of model parameter uncertainty in the estimation process further impaired the identifiability of initial pools, especially for the more slowly decomposing pools. Our illustrative case studies show how Bayesian inverse modeling can be used to provide important insights into the information content of incubation experiments. Moreover, the outcome of this virtual experiment helps to explain the results of related real-world studies on SOC dynamics.

Evaluation of Methods to Improve the Extraction and Recovery of DNA from Cotton Swabs for Forensic Analysis

PubMed Central

Adamowicz, Michael S.; Stasulli, Dominique M.; Sobestanovich, Emily M.; Bille, Todd W.

2014-01-01

Samples for forensic DNA analysis are often collected from a wide variety of objects using cotton or nylon tipped swabs. Testing has shown that significant quantities of DNA are retained on the swab, however, and subsequently lost. When processing evidentiary samples, the recovery of the maximum amount of available DNA is critical, potentially dictating whether a usable profile can be derived from a piece of evidence or not. The QIAamp DNA Investigator extraction kit was used with its recommended protocol for swabs (one hour incubation at 56°C) as a baseline. Results indicate that over 50% of the recoverable DNA may be retained on the cotton swab tip, or otherwise lost, for both blood and buccal cell samples when using this protocol. The protocol’s incubation time and temperature were altered, as was incubating while shaking or stationary to test for increases in recovery efficiency. An additional step was then tested that included periodic re-suspension of the swab tip in the extraction buffer during incubation. Aliquots of liquid blood or a buccal cell suspension were deposited and dried on cotton swabs and compared with swab-less controls. The concentration of DNA in each extract was quantified and STR analysis was performed to assess the quality of the extracted DNA. Stationary incubations and those performed at 65°C did not result in significant gains in DNA yield. Samples incubated for 24 hours yielded less DNA. Increased yields were observed with three and 18 hour incubation periods. Increases in DNA yields were also observed using a swab re-suspension method for both cell types. The swab re-suspension method yielded an average two-fold increase in recovered DNA yield with buccal cells and an average three-fold increase with blood cells. These findings demonstrate that more of the DNA collected on swabs can be recovered with specific protocol alterations. PMID:25549111

Anaerobic aquifer transformations of 2,4-Dinitrophenol under different terminal electron accepting conditions

USGS Publications Warehouse

Krumholz, L.R.; Suflita, J.M.

1997-01-01

We evaluated the susceptibility of 2,4-dinitrophenol (2,4-DNP) and 2,4-diaminophenol to anaerobic biodegradation in aquifer slurries. Aquifer microorganisms depleted 2,4-DNP at rates of 25, 9 and 0.4 μM/day under methanogenic, sulfate-reducing and nitrate-reducing conditions, respectively. Rates of abiotic, 2,4-DNP loss in autoclaved control incubations were 7.2, 6.2 and 0.95 μM/day respectively. Abiotic, 2,4-DNP reduction was especially important as the first step in its transformation. 2-Amino-4-nitrophenol was produced by this process, but this compound was further metabolized in methanogenic and sulfate-reducing aquifer slurries. This partially reduced compound persisted in autoclaved controls and in the nitrate-reducing aquifer slurries. Aquifer slurries incubated with either 2,4-DNP or 2,4-diaminophenol produced methane when incubated with no other electron acceptor suggesting that mineralization had occurred under these conditions. In parallel experiments, aquifer slurries amended with 2,6-dinitrophenol or picric acid did not produce methane at levels above the substrate unamended controls.

Chemical Compositions and Aroma Evaluation of Volatile Oil from the Industrial Cultivation Medium of Enterococcus faecalis.

PubMed

Ono, Toshirou; Usami, Atsushi; Nakaya, Satoshi; Maeba, Keisuke; Yonejima, Yasunori; Toyoda, Masanori; Ikeda, Atsushi; Miyazawa, Mitsuo

2015-01-01

Enterococcus faecalis is one of the major lactic acid bacterium (LAB) species colonizing the intestines of animals and humans. The characteristic odor of the volatile oils obtained from both the liquid medium after incubation (MAI) and liquid medium before incubation (MBI) in the cultivation process of E. faecalis was investigated to determine the utility of the liquid medium. In total, fifty-six and thirty-two compounds were detected in the volatile oils from the MAI (MAI oil) and MBI (MBI oil), respectively. The principle components of MAI oil were 2,5-dimethylpyrazine (19.3%), phenylacetaldehyde (19.3%), and phenylethyl alcohol (9.3%). The aroma extract dilution analysis (AEDA) method was performed using gas chromatography-olfactometry (GC-O). The total number of aroma-active compounds identified in the volatile oil from MBI and MAI was thirteen compounds; in particular, 5-methyl-2-furanmethanol, phenylacetaldehyde, and phenylethyl alcohol were the most primary aroma-active compounds in MAI oil. These results imply that the industrial cultivation medium after incubation of E. faecalis may be utilized as a source of volatile oils.

Initial growth processes in the epitaxy of Ge with GeH{sub 4} on oxidized Si substrates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Angermeier, D.; Kuhn, W.S.; Druihle, R.

1997-02-01

The heteroepitaxial growth of Ge on (100) Si in a horizontal, atmospheric pressure metallorganic vapor-phase epitaxy reactor is reported using germane GeH{sub 4} (0.1% in H{sub 2}). A particularly crucial parameter for germanium deposition on silicon is the time for the onset of epitaxial growth, the incubation time. The time was measured at substrate temperatures between 450 and 600{degree}C. At a substrate temperature of 450{degree}C an incubation time of 520 s was found and for the subsequent epitaxy growth rates of 50 nm/min were determined by Nomarski microscopy and electron diffraction. The existence of residual oxide in the reactor chambermore » forming an in situ SiO{sub 2} layer was evaluated by x-ray photoemission spectroscopy. To obtain a more thorough understanding of the gas- and solid-phase composition of Ge, Si, and oxygen the Gibbs energy of the system was calculated for various growth temperatures. It was concluded that SiO{sub 2} molecules are reduced by GeH{sub 4} molecules during the incubation period.« less

Optimization of an incubation step to maximize sulforaphane content in pre-processed broccoli.

PubMed

Mahn, Andrea; Pérez, Carmen

2016-11-01

Sulforaphane is a powerful anticancer compound, found naturally in food, which comes from the hydrolysis of glucoraphanin, the main glucosinolate of broccoli. The aim of this work was to maximize sulforaphane content in broccoli by designing an incubation step after subjecting broccoli pieces to an optimized blanching step. Incubation was optimized through a Box-Behnken design using ascorbic acid concentration, incubation temperature and incubation time as factors. The optimal incubation conditions were 38 °C for 3 h and 0.22 mg ascorbic acid per g fresh broccoli. The maximum sulforaphane concentration predicted by the model was 8.0 µmol g -1 , which was confirmed experimentally yielding a value of 8.1 ± 0.3 µmol g -1 . This represents a 585% increase with respect to fresh broccoli and a 119% increase in relation to blanched broccoli, equivalent to a conversion of 94% of glucoraphanin. The process proposed here allows maximizing sulforaphane content, thus avoiding artificial chemical synthesis. The compound could probably be isolated from broccoli, and may find application as nutraceutical or functional ingredient.

Incubation and Intuition in Creative Problem Solving.

PubMed

Gilhooly, Kenneth J

2016-01-01

Creative problem solving, in which novel solutions are required, has often been seen as involving a special role for unconscious processes (Unconscious Work) which can lead to sudden intuitive solutions (insights) when a problem is set aside during incubation periods. This notion of Unconscious Work during incubation periods is supported by a review of experimental studies and particularly by studies using the Immediate Incubation paradigm. Other explanations for incubation effects, in terms of Intermittent Work or Beneficial Forgetting are considered. Some recent studies of divergent thinking, using the Alternative Uses task, carried out in my laboratory regarding Immediate vs. Delayed Incubation and the effects of resource competition from interpolated activities are discussed. These studies supported a role for Unconscious Work as against Intermittent Conscious work or Beneficial Forgetting in incubation.

Incubation and Intuition in Creative Problem Solving

PubMed Central

Gilhooly, Kenneth J.

2016-01-01

Creative problem solving, in which novel solutions are required, has often been seen as involving a special role for unconscious processes (Unconscious Work) which can lead to sudden intuitive solutions (insights) when a problem is set aside during incubation periods. This notion of Unconscious Work during incubation periods is supported by a review of experimental studies and particularly by studies using the Immediate Incubation paradigm. Other explanations for incubation effects, in terms of Intermittent Work or Beneficial Forgetting are considered. Some recent studies of divergent thinking, using the Alternative Uses task, carried out in my laboratory regarding Immediate vs. Delayed Incubation and the effects of resource competition from interpolated activities are discussed. These studies supported a role for Unconscious Work as against Intermittent Conscious work or Beneficial Forgetting in incubation. PMID:27499745

High Pressure Serpentinization Catalysed by Awaruite in Planetary Bodies

NASA Astrophysics Data System (ADS)

Neto-Lima, J.; Fernández-Sampedro, M.; Prieto-Ballesteros, O.

2017-10-01

Recent discoveries from planetary missions show that serpentinization process may act significantly on the geological evolution and potential habitability of the icy bodies of the Solar System, like Enceladus or Europa. Here we review the available experimental data so far about methane formation occurring during serpentinization, which is potentially relevant to icy moons, and present our results using awaruite as a catalyst of this process. The efficiency of awaruite and high pressure in the Fischer-Tropsch and Sabatier Type reactions are evaluated here when olivine is incubated.

Co-occurring anammox, denitrification, and codenitrification in agricultural soils.

PubMed

Long, Andrew; Heitman, Joshua; Tobias, Craig; Philips, Rebecca; Song, Bongkeun

2013-01-01

Anammox and denitrification mediated by bacteria are known to be the major microbial processes converting fixed N to N(2) gas in various ecosystems. Codenitrification and denitrification by fungi are additional pathways producing N(2) in soils. However, fungal codenitrification and denitrification have not been well investigated in agricultural soils. To evaluate bacterial and fungal processes contributing to N(2) production, molecular and (15)N isotope analyses were conducted with soil samples collected at six different agricultural fields in the United States. Denitrifying and anammox bacterial abundances were measured based on quantitative PCR (qPCR) of nitrous oxide reductase (nosZ) and hydrazine oxidase (hzo) genes, respectively, while the internal transcribed spacer (ITS) of Fusarium oxysporum was quantified to estimate the abundance of codenitrifying and denitrifying fungi. (15)N tracer incubation experiments with (15)NO(3)(-) or (15)NH(4)(+) addition were conducted to measure the N(2) production rates from anammox, denitrification, and codenitrification. Soil incubation experiments with antibiotic treatments were also used to differentiate between fungal and bacterial N(2) production rates in soil samples. Denitrifying bacteria were found to be the most abundant, followed by F. oxysporum based on the qPCR assays. The potential denitrification rates by bacteria and fungi ranged from 4.118 to 42.121 nmol N(2)-N g(-1) day(-1), while the combined potential rates of anammox and codenitrification ranged from 2.796 to 147.711 nmol N(2)-N g(-1) day(-1). Soil incubation experiments with antibiotics indicated that fungal codenitrification was the primary process contributing to N(2) production in the North Carolina soil. This study clearly demonstrates the importance of fungal processes in the agricultural N cycle.

Effects of egg quality and method of incubation on the hatching success of channel X blue hybrid catfish eggs

USDA-ARS?s Scientific Manuscript database

The objective of this study was to evaluate the effects of egg quality of stripped eggs from channel catfish (Ictalurus punctatus) and method of incubation of fertilized hybrid catfish eggs on hatching success. Stripped eggs from 17 channel catfish females were evaluated in a 2 x 2 factorial...

Effect of the holding time at 15 °C prior to cryopreservation, the thawing rate and the post-thaw incubation temperature on the boar sperm quality after cryopreservation.

PubMed

Tomás, Cristina; Gómez-Fernández, José; Gómez-Izquierdo, Emilio; de Mercado, Eduardo

2014-01-30

The aim of the present study was to evaluate the effect of the holding time at 15 °C prior to cryopreservation (2, 4 and 8h), thawing rate (37 °C for 20s or 70 °C for 8s) and post-thaw incubation temperature (15 °C or 37 °C) on the post-thaw boar sperm quality. These are important time periods in the freezing-thawing process which have been less studied. Sperm-rich ejaculate fractions from three healthy boars were collected once a week for five consecutive weeks and were cryopreserved with the lactose-egg yolk extender (LEY). Sperm quality was determined by assessing the motility, the acrosome status, and the sperm plasma membrane integrity at 30, 150 and 240 min of incubation. The results show that with the holding time at 15 °C prior to cryopreservation there was not a clear effect until at least 24h of holding time. The thawing rate and the post-thaw incubation temperature, however, had a marked effect on sperm quality. When the samples were thawed at 70 °C for 8s, the sperm viability, motility and some kinetic variables (VCL, VSL, VAP and ALH) were greater than with results observed when the samples were thawed at 37 °C for 20s. In addition after thawing the sperm samples incubated at 15 °C had a sustained sperm quality for longer, up to 4h post-thawing. Copyright © 2014 Elsevier B.V. All rights reserved.

The effect of curcumin on insulin release in rat-isolated pancreatic islets.

PubMed

Abdel Aziz, Mohamed T; El-Asmar, Mohamed F; El Nadi, Essam G; Wassef, Mohamed A; Ahmed, Hanan H; Rashed, Laila A; Obaia, Eman M; Sabry, Dina; Hassouna, Amira A; Abdel Aziz, Ahmed T

2010-08-01

Curcumin exerts a hypoglycemic action and induces heme-oxygenase-1 (HO-1). We evaluated the effect of curcumin on isolated islets of Langerhans and studied whether its action on insulin secretion is mediated by inducible HO-1. Islets were isolated from rats and divided into control islets, islets incubated in different curcumin concentrations, islets incubated in hemin, islets incubated in curcumin and HO inhibitor, stannous mesoporphyrin (SnMP), islets incubated in hemin and SnMP, islets incubated in SnMP only, and islets incubated in 16.7 mmol/L glucose. Heme-oxygenase activity, HO-1 expression, and insulin estimation was assessed. Insulin secretion, HO-1 gene expression and HO activity were significantly increased in islets incubated in curcumin, hemin, and glucose compared with controls. This increase in insulin secretion was significantly decreased by incubation of islets in SnMP. The action of curcumin on insulin secretion from the isolated islets may be, in part, mediated through increased HO-1 gene expression.

Evaluation of protective effect of amifostine on dacarbazine induced genotoxicity.

PubMed

Etebari, M; Jafarian-Dehkordi, A; Lame, V

2015-01-01

Anticancer therapy with alkylating agents has been used for many years. Dacarbazine (DTIC) as an alkylating agent is used alone or in combination with other chemotherapy drugs. In order to inhibit the formation of secondary cancers resulting from chemotherapy with DTIC, preventional strategies is necessary. The present study was undertaken to evaluate the genoprotective effect of amifostine on the genotoxic effects of DTIC in cell culture condition. To determine the optimum genotoxic concentration of DTIC, HepG2 cells were incubated with various DTIC concentrations including 5, 10 and 20 μg/ml for 2 h and the genotoxic effects were evaluated by the comet assay. The result of this part of the study showed that incubation of HepG2 cells with DTIC at 5 μg/ml was sufficient to produce genotoxic effect. In order to determine the protective effects of amifostine on genotoxicity induced by DTIC, HepG2 cells were incubated with different concentrations of amifostine (2, 3 and 5 mg/ml) for 1 h which was followed by incubation with DTIC at 5 μg/ml for 2 h. One hour incubation of cells with different concentrations of amifostine before incubation with DITC indicated that at least 5 mg/ml concentration of amifostine can prevent genotoxic effects induced by DTIC on HepG2 cells under described condition. In conclusion amifostine could prevent DNA damage induced by DTIC on HepG2 cells.

Enzyme activity and AGE formation in a model of AST glycoxidation by D-fructose in vitro.

PubMed

Bousova, Iva; Vukasović, Danka; Juretić, Dubravka; Palicka, Vladimir; Drsata, Jaroslav

2005-03-01

Non-enzymatic glycation as the chain reaction between reducing sugars and free amino groups of proteins has been shown to correlate with physiological ageing and severity of diabetes. The process involves oxidative steps (glycoxidation). In this paper, the effect of D-fructose as a reactive sugar on aspartate aminotransferase (AST) as a model protein was monitored by measurements of the enzyme activity and formation of fluorescent advanced glycation end products (AGEs). Change in the AST activity was considered as a measure of the overall protein damage caused by glycation, and total AGEs and pentosidine represent, at least partly, the formation of glycoxidation products. Catalytic activity of AST in an incubation mixture containing D-fructose (50 mmol L(-1)), decreased compared to control values to 42% (p < 0.05) and to 11% (p < 0.05) on the 5th and on 21st day of incubation, respectively. In the presence of fructose, total fluorescent AGEs concentration was significantly higher since 5th day of incubation (110%, p < 0.05) and the fluorescent pentosidine concentration from 15th day of incubation (117%, p < 0.05) compared to control values, respectively. Catalytic activity of AST clearly and quantitatively demonstrated functional changes in the enzyme molecule caused by structural modifications initiated by fructose, while the evaluation of AGE formation and especially that of pentosidine by fluorescence measurement was less reliable.

Preliminary data on growth and enzymatic abilities of soil fungus Humicolopsis cephalosporioides at different incubation temperatures.

PubMed

Elíades, Lorena Alejandra; Cabello, Marta N; Pancotto, Verónica; Moretto, Alicia; Rago, María Melisa; Saparrat, Mario C N

2015-01-01

Nothofagus pumilio (Poepp & Endl.) Krasser, known as "lenga" is the most important timber wood species in southernmost Patagonia (Argentina). Humicolopsis cephalosporioides Cabral & Marchand is a soil fungus associated with Nothofagus pumilio forests, which has outstanding cellulolytic activity. However, there is no information about the ability of this fungus to use organic substrates other than cellulose, and its ability to produce different enzyme systems, as well as its response to temperature. The aim of this study was to examine the role of H. cephalosporioides in degradation processes in N. pumilio forests in detail by evaluating the in vitro ability of four isolates of this fungus to grow and produce different lytic enzyme systems, and their response to incubation temperature. The ability of the fungi to grow and produce enzyme systems was estimated by inoculating them on agar media with specific substrates, and the cultures were incubated at three temperatures. A differential behavior of each strain in levels of growth and enzyme activity was found according to the medium type and/or incubation temperature. A intra-specific variability was found in H. cephalosporioides. Likewise a possible link between the saprotrophic role of this fungus in N. pumilio forests and the degradation of organic matter under stress conditions, such as those from frosty environments, was also discussed. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

The strain-encoded relationship between PrP replication, stability and processing in neurons is predictive of the incubation period of disease.

PubMed

Ayers, Jacob I; Schutt, Charles R; Shikiya, Ronald A; Aguzzi, Adriano; Kincaid, Anthony E; Bartz, Jason C

2011-03-01

Prion strains are characterized by differences in the outcome of disease, most notably incubation period and neuropathological features. While it is established that the disease specific isoform of the prion protein, PrP(Sc), is an essential component of the infectious agent, the strain-specific relationship between PrP(Sc) properties and the biological features of the resulting disease is not clear. To investigate this relationship, we examined the amplification efficiency and conformational stability of PrP(Sc) from eight hamster-adapted prion strains and compared it to the resulting incubation period of disease and processing of PrP(Sc) in neurons and glia. We found that short incubation period strains were characterized by more efficient PrP(Sc) amplification and higher PrP(Sc) conformational stabilities compared to long incubation period strains. In the CNS, the short incubation period strains were characterized by the accumulation of N-terminally truncated PrP(Sc) in the soma of neurons, astrocytes and microglia in contrast to long incubation period strains where PrP(Sc) did not accumulate to detectable levels in the soma of neurons but was detected in glia similar to short incubation period strains. These results are inconsistent with the hypothesis that a decrease in conformational stability results in a corresponding increase in replication efficiency and suggest that glia mediated neurodegeneration results in longer survival times compared to direct replication of PrP(Sc) in neurons.

Fate of Escherichia coli O157 Cells Inoculated into Lightly Pickled Chinese Cabbage during Processing, Storage and Incubation in Artificial Gastric Juice.

PubMed

Inatsu, Yasuhiro; Ohata, Yukiko; Ananchaipattana, Chiraporn; Latiful Bari, Md; Hosotani, Yukie; Kawasaki, Susumu

2016-01-01

Fate of Escherichia coli O157 cells was evaluated when inoculated into each step after production of lightly pickled Chinese cabbage. The efficacy of surface sterilization by 100 mg/L of chlorine water for 10 min on raw leaves (6.0 log CFU/g) was 2.2 log CFU/g reduction. No meaningful change of the population of E. coli O157 (3.5 log CFU/g to 1.5 log MPN/g) contaminated into 19 kinds of products was observed. These results indicated the difficulty of estimating the viable count of the cells between contaminated on farms and further processing and storage steps. The population of E. coli O157 (3 log CFU/g to 1 log MPN/g) inoculated into the Chinese cabbage products was reduced less than 0.6 log CFU/g after 2 h-incubation at 37℃ in artificial gastric juice. Prevention from initial contamination of E. coli O157 on the ingredients of Chinese cabbage products is important to reduce the risk of food poisoning because the reduction of the bacterial counts after processing and consumption are limited.

Reproducing stone monument photosynthetic-based colonization under laboratory conditions.

PubMed

Miller, Ana Zélia; Laiz, Leonila; Gonzalez, Juan Miguel; Dionísio, Amélia; Macedo, Maria Filomena; Saiz-Jimenez, Cesareo

2008-11-01

In order to understand the biodeterioration process occurring on stone monuments, we analyzed the microbial communities involved in these processes and studied their ability to colonize stones under controlled laboratory experiments. In this study, a natural green biofilm from a limestone monument was cultivated, inoculated on stone probes of the same lithotype and incubated in a laboratory chamber. This incubation system, which exposes stone samples to intermittently sprinkling water, allowed the development of photosynthetic biofilms similar to those occurring on stone monuments. Denaturing gradient gel electrophoresis (DGGE) analysis was used to evaluate the major microbial components of the laboratory biofilms. Cyanobacteria, green microalgae, bacteria and fungi were identified by DNA-based molecular analysis targeting the 16S and 18S ribosomal RNA genes. The natural green biofilm was mainly composed by the Chlorophyta Chlorella, Stichococcus, and Trebouxia, and by Cyanobacteria belonging to the genera Leptolyngbya and Pleurocapsa. A number of bacteria belonging to Alphaproteobacteria, Bacteroidetes and Verrucomicrobia were identified, as well as fungi from the Ascomycota. The laboratory colonization experiment on stone probes showed a colonization pattern similar to that occurring on stone monuments. The methodology described in this paper allowed to reproduce a colonization equivalent to the natural biodeteriorating process.

Experimental investigation on phase change materials as heating element for non-electric neonatal incubator

NASA Astrophysics Data System (ADS)

Matahari, Rho Natta; Putra, Nandy; Ariantara, Bambang; Amin, Muhammad; Prawiro, Erwin

2017-02-01

High number of preterm births is one of the issues in improving health standard. The effort to help premature babies is hampered by high cost of NICU care in hospital. In addition, uneven distribution of electricity to remote area made it hard to operate the incubator. Utilization of phase change material beeswax to non-electricity incubator as heating element becomes alternative option to save premature babies. The objective of this experiment is to investigate the most efficient mass of beeswax according to Indonesian National Standard to earn over time and ideal temperature of incubator. Experiment was performed using prototype incubator, which utilizes natural convection phenomenon in the heating process of incubator. Utilization of fin is to accelerate heat distribution in the incubator. Result of experiment showed that the most efficient mass of PCM is 3 kg, which has 2.45 hours of running time for maintaining temperature of incubator in range of 32-36 °C.

Effects of environmental factors on incubation patterns of Greater Sage-Grouse

USGS Publications Warehouse

Coates, Peter S.; Delehanty, David J.

2008-01-01

Birds in which only one sex incubates the eggs are often faced with a direct conflict between foraging to meet metabolic needs and incubation. Knowledge of environmental and ecological factors that shape life-history strategies of incubation is limited. We used continuous videography to make precise measurements of female Greater Sage-Grouse (Centrocercus urophasianus) incubation constancy (percentage of time spent at the nest in a 24-hour period) and recess duration. We used an information-theoretic approach to evaluate incubation patterns in relation to grouse age, timing of incubation, raven abundance, microhabitat, weather, and food availability. Overall, sage-grouse females showed an incubation constancy of 96% and a distinctive bimodal distribution of brief incubation recesses that peaked at sunset and 30 min prior to sunrise. Grouse typically returned to their nests during low light conditions. Incubation constancy of yearlings was lower than that of adults, particularly in the later stages of incubation. Yearlings spent more time away from nests later in the morning and earlier in the evening compared to adults. Video images revealed that nearly all predation events by Common Ravens (Corvus corax), the most frequently recorded predator at sage-grouse nests, took place during mornings and evenings after sunrise and before sunset, respectively. These were the times of the day when sage-grouse typically returned from incubation recesses. Recess duration was negatively related to raven abundance. We found evidence that incubation constancy increased with greater visual obstruction, usually from vegetation, of nests. An understanding of how incubation patterns relate to environmental factors will help managers make decisions aimed at increasing productivity through successful incubation.

A Study of Business Incubators: Models, Best Practices, and Recommendations for NASA and Florida

NASA Technical Reports Server (NTRS)

1997-01-01

This study was conducted to provide NASA-Kennedy Space Center with information and recommendations to support establishing one or more technology-based business incubators In Florida. The study involved assembling information about incubators: why they succeed, why they fail, how they are organized, and what services they provide. Consequently, this study focuses on widely-recognized "best practices," needed to establish successful technology- based business incubators. The findings are used to optimize the design and implementation of one or more technology-based business incubators to be established in Florida. Recommendations reflect both the essential characteristics of successful incubators and the optimal business demographics in Florida. Appendix A provides a fuller description of the objectives of the study. Technology-based business incubators are an increasing catalyst of new business development across the USi Incubators focus on providing entrepreneurs and small start-up firms with a wide array of support services necessary to bring forth new products and processes based on technologies developed in the nation's federal and private laboratories and universities. Appendix B provides extensive discussion of findings relative to technology- based business incubators.

21 CFR 640.81 - Processing.

Code of Federal Regulations, 2013 CFR

2013-04-01

... product as referred to in § 640.84(d). (g) Incubation. All final containers of Albumin (Human) shall be... this section. At the end of this incubation period, each final container shall be examined and all...

21 CFR 640.81 - Processing.

Code of Federal Regulations, 2012 CFR

2012-04-01

... product as referred to in § 640.84(d). (g) Incubation. All final containers of Albumin (Human) shall be... this section. At the end of this incubation period, each final container shall be examined and all...

21 CFR 640.81 - Processing.

Code of Federal Regulations, 2014 CFR

2014-04-01

... product as referred to in § 640.84(d). (g) Incubation. All final containers of Albumin (Human) shall be... this section. At the end of this incubation period, each final container shall be examined and all...

Caspase activation, hydrogen peroxide production and Akt dephosphorylation occur during stallion sperm senescence.

PubMed

Gallardo Bolaños, J M; Balao da Silva, C; Martín Muñoz, P; Plaza Dávila, M; Ezquerra, J; Aparicio, I M; Tapia, J A; Ortega Ferrusola, C; Peña, F J

2014-08-01

To investigate the mechanisms inducing sperm death after ejaculation, stallion ejaculates were incubated in BWW media during 6 h at 37°C. At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeability and integrity were evaluated (flow cytometry). Also, at the same time intervals, active caspase 3, hydrogen peroxide, superoxide anion (flow cytometry) and Akt phosphorylation (flow cytometry) were evaluated. Major decreases in sperm function occurred after 6 h of incubation, although after 1 h decrease in the percentages of motile and progressive motile sperm occurred. The decrease observed in sperm functionality after 6 h of incubation was accompanied by a significant increase in the production of hydrogen peroxide and the greatest increase in caspase 3 activity. Additionally, the percentage of phosphorylated Akt reached a minimum after 6 h of incubation. These results provide evidences that sperm death during in vitro incubation is largely an apoptotic phenomena, probably stimulated by endogenous production of hydrogen peroxide and the lack of prosurvival factors maintaining Akt in a phosphorylated status. Disclosing molecular mechanisms leading to sperm death may help to develop new strategies for stallion sperm conservation. © 2014 Blackwell Verlag GmbH.

Cleanrooms and tissue banking how happy I could be with either GMP or GTP?

PubMed

Klykens, J; Pirnay, J-P; Verbeken, G; Giet, O; Baudoux, E; Jashari, R; Vanderkelen, A; Ectors, N

2013-12-01

The regulatory framework of tissue banking introduces a number of requirements for monitoring cleanrooms for processing tissue or cell grafts. Although a number of requirements were clearly defined, some requirements are open for interpretation. This study aims to contribute to the interpretation of GMP or GTP guidelines for tissue banking. Based on the experience of the participating centers, the results of the monitoring program were evaluated to determine the feasibility of a cleanroom in tissue banking and the monitoring program. Also the microbial efficacy of a laminar airflow cabinet and an incubator in a cleanroom environment was evaluated. This study indicated that a monitoring program of a cleanroom at rest in combination with (final) product testing is a feasible approach. Although no statistical significance (0.90 < p < 0.95) was found there is a strong indication that a Grade D environment is not the ideal background environment for a Grade A obtained through a laminar airflow cabinet. The microbial contamination of an incubator in a cleanroom is limited but requires closed containers for tissue and cell products.

Erythrocytes L-arginine y+ transporter inhibition by N-ethylmaleimide in ice-bath.

PubMed

Pinheiro da Costa, Bartira Ercília; de Almeida, Priscilla Barcellos; Conceição, Ioná Rosine; Antonello, Ivan Carlos Ferreira; d'Avila, Domingos O; Poli-de-Figueiredo, Carlos Eduardo

2010-11-01

Erythrocytes L: -arginine uptake is conveyed by y+ and y+L membrane transport systems. Pre-incubation with N-ethylmaleimide for 10 min at 37°C inhibits the y+ system. The aim of this study was to determine the ideal pre-incubation temperature in evaluating y+ and y+L systems. Cells were pre-incubated with or without N-ethylmaleimide for 10 min at 4°C and 37°C. L: -Arginine uptake was quantified by radioisotope and standard erythrocytes membrane flux methodology. Results demonstrate that erythrocytes L: -arginine content is depleted by pre-incubation at 37°C for 10 min, thus changing the V (max) measurement. The inhibitory effect of N-ethylmaleimide pre-incubation was temperature independent and already complete after 1 min of incubation. No significant difference in kinetic parameters was detected between cells pre-incubated at 37°C or 4°C, under zero-trans conditions. In conclusion, we suggest that measurement of erythrocytes L: -arginine uptake by y+ and y+L systems could be carried out without N-ethylmaleimide pre-incubation at 37°C.

Enzymatic added extraction and clarification of fruit juices-A review.

PubMed

Sharma, Harsh P; Patel, Hiral; Sugandha

2017-04-13

Enzymatic treatment for juice extraction is most commonly used now a days. The enzymatic process is claimed to offer a number of advantages over mechanical-thermal comminution of several fruit pulps. Enzymes are an integral component of modern fruit juice manufacturing and are highly suitable for optimizing processes. Their main purposes are: increase extraction of juice from raw material, increase processing efficiency (pressing, solid settling or removal), and generate a final product that is clear and visually attractive. Juice extraction can be done by using various mechanical processes, which may be achieved through diffusion extraction, decanter centrifuge, screw type juice extractor, fruit pulper and by different types of presses. Enzymatic treatment prior to mechanical extraction significantly improves juice recovery compared to any other extraction process. Enzymatic hydrolysis of the cell walls increases the extraction yield, reducing sugars, soluble dry matter content and galacturonic acid content and titrable acidity of the products. Enzymatic degradation of the biomaterial depends upon the type of enzyme, incubation time, incubation temperature, enzyme concentration, agitation, pH and use of different enzyme combinations. We can conclude from the technical literature that use of the enzymes i.e. cellulases, pectinases, amylases and combination of these enzymes can give better juice yield with superior quality of the fruit juice. Pectinase enzyme can give maximum juice yield i.e. 92.4% at 360 minutes incubation time, 37°C incubation temperature and 5 mg/100 g of enzyme concentration. Whereas the combination of two enzymes i.e. pectin methyl esterase (PME) and polygalacturonase (PG) at 120 minutes of incubation time, 50°C of incubation temperature and 0.05 mg/100 gm of enzymatic concentration can give the maximum yield of 96.8% for plum fruits. This paper discusses the use of enzymes in fruit juice production focusing on the juice recovery, clarity and effect of the particular enzyme on the biochemical properties of the fruit juices.

Neutral temperature range in incubators: performance of equipment in current use and new developments.

PubMed

Libert, J P; Bach, V; Farges, G

1997-01-01

Low-birth-weight neonates should be nursed at thermoneutrality inside incubators. Thermoneutrality control is essential to enhance body growth and to reduce neonatal illnesses and mortality. Guidelines have been published to provide the thermoneutral range, but the recommendations did not always take into account all ambient and physiological parameters influencing thermoneutrality. In most marketed incubators, the heat supply is controlled through convective air flow (closed incubators) or through radiant power density (radiant warmer beds). The heating unit (on/off cycling or adjustable proportional control) is activated by an error signal calculated from the difference between a controlled temperature and a reference value preset by the clinician. The controlled variable can be either the incubator air or the skin temperature of the anterior abdominal region of the neonate. The neonate's size, thermal properties of the mattress and of incubator walls, air temperature and humidity, air velocity, incubator wall temperatures all influence the heat exchanges between the neonate and the surroundings, and, consequently, modify the obtention of thermoneutrality. Moreover, studies of the physiological mechanisms by which the neonate regulates body heat storage suggest that metabolic rate, behavior, vigilance level, nursing care, and heater control processes should also be taken into account. Little attention has been paid to these factors, and incubator performances are often disappointing. This article reviews the different factors that modify thermoneutral condition. An attempt is made to suggest new ways to design equipment incorporating these factors in algorithms controlling heater processes in order to reach the optimal thermal environment in which the neonate should be nursed.

Light scattering method to measure red blood cell aggregation during incubation

NASA Astrophysics Data System (ADS)

Grzegorzewski, B.; Szołna-Chodór, A.; Baryła, J.; DreŻek, D.

2018-01-01

Red blood cell (RBC) aggregation can be observed both in vivo as well as in vitro. This process is a cause of alterations of blood flow in microvascular network. Enhanced RBC aggregation makes oxygen and nutrients delivery difficult. Measurements of RBC aggregation usually give a description of the process for a sample where the state of a solution and cells is well-defined and the system reached an equilibrium. Incubation of RBCs in various solutions is frequently used to study the effects of the solutions on the RBC aggregation. The aggregation parameters are compared before and after incubation while the detailed changes of the parameters during incubation remain unknown. In this paper we have proposed a method to measure red blood cell aggregation during incubation based on the well-known technique where backscattered light is used to assess the parameters of the RBC aggregation. Couette system consisting of two cylinders is adopted in the method. The incubation is observed in the Couette system. In the proposed method following sequence of rotations is adapted. Two minutes rotation is followed by two minutes stop. In this way we have obtained a time series of back scattered intensity consisting of signals respective for disaggregation and aggregation. It is shown that the temporal changes of the intensity manifest changes of RBC aggregation during incubation. To show the ability of the method to assess the effect of incubation time on RBC aggregation the results are shown for solutions that cause an increase of RBC aggregation as well as for the case where the aggregation is decreased.

Effects of incubation temperature on growth and performance of the veiled chameleon (Chamaeleo calyptratus).

PubMed

Andrews, Robin M

2008-10-01

I evaluated the effect of incubation temperature on phenotypes of the veiled chameleon, Chamaeleo calyptratus. I chose this species for study because its large clutch size (30-40 eggs or more) allows replication within clutches both within and among experimental treatments. The major research objectives were (1) to assess the effect of constant low, moderate, and high temperatures on embryonic development, (2) to determine whether the best incubation temperature for embryonic development also produced the "best" hatchlings, and (3) to determine how a change in incubation temperature during mid-development would affect phenotype. To meet these objectives, I established five experimental temperature regimes and determined egg survival and incubation length and measured body size and shape, selected body temperatures, and locomotory performance of lizards at regular intervals from hatching to 90 d, or just before sexual maturity. Incubation temperature affected the length of incubation, egg survival, and body mass, but did not affect sprint speed or selected body temperature although selected body temperature affected growth in mass independently of treatment and clutch. Incubation at moderate temperatures provided the best conditions for both embryonic and post-hatching development. The highest incubation temperatures were disruptive to development; eggs had high mortality, developmental rate was low, and hatchlings grew slowly. Changes in temperature during incubation increased the among-clutch variance in incubation length relative to that of constant temperature treatments. Copyright 2008 Wiley-Liss, Inc.

Argon attenuates the emergence of secondary injury after traumatic brain injury within a 2-hour incubation period compared to desflurane: an in vitro study.

PubMed

Grüßer, Linda; Blaumeiser-Debarry, Rosmarie; Krings, Matthias; Kremer, Benedikt; Höllig, Anke; Rossaint, Rolf; Coburn, Mark

2017-01-01

Despite years of research, treatment of traumatic brain injury (TBI) remains challenging. Considerable data exists that some volatile anesthetics might be neuroprotective. However, several studies have also revealed a rather neurotoxic profile of anesthetics. In this study, we investigated the effects of argon 50%, desflurane 6% and their combination in an in vitro TBI model with incubation times similar to narcotic time slots in a daily clinical routine. Organotypic hippocampal brain slices of 5- to 7-day-old mice were cultivated for 14 days before TBI was performed. Slices were eventually incubated for 2 hours in an atmosphere containing no anesthetic gas, argon 50% or desflurane 6% or both. Trauma intensity was evaluated via fluorescent imagery. Our results show that neither argon 50% nor desflurane 6% nor their combination could significantly reduce the trauma intensity in comparison to the standard atmosphere. However, in comparison to desflurane 6%, argon 50% displayed a rather neuroprotective profile within the first 2 hours after a focal mechanical trauma ( P = 0.015). A 2-hour incubation in an atmosphere containing both gases, argon 50% and desflurane 6%, did not result in significant effects in comparison to the argon 50% group or the desflurane 6% group. Our findings demonstrate that within a 2-hour incubation time neither argon nor desflurane could affect propidium iodide-detectable cell death in an in vitro TBI model in comparison to the standard atmosphere, although cell death was less with argon 50% than with desflurane 6%. The results show that within this short time period processes concerning the development of secondary injury are already taking place and may be manipulated by argon.

Release kinetics of platelet-derived and plasma-derived growth factors from autologous plasma rich in growth factors.

PubMed

Anitua, Eduardo; Zalduendo, Mari Mar; Alkhraisat, Mohammad Hamdan; Orive, Gorka

2013-10-01

Many studies have evaluated the biological effects of platelet rich plasma reporting the final outcomes on cell and tissues. However, few studies have dealt with the kinetics of growth factor delivery by plasma rich in growth factors. Venous blood was obtained from three healthy volunteers and processed with PRGF-Endoret technology to prepare autologous plasma rich in growth factors. The gel-like fibrin scaffolds were then incubated in triplicate, in a cell culture medium to monitor the release of PDGF-AB, VEGF, HGF and IGF-I during 8 days of incubation. A leukocyte-platelet rich plasma was prepared employing the same technology and the concentrations of growth factors and interleukin-1β were determined after 24h of incubation. After each period, the medium was collected, fibrin clot was destroyed and the supernatants were stored at -80°C until analysis. The growth factor delivery is diffusion controlled with a rapid initial release by 30% of the bioactive content after 1h of incubation and a steady state release when almost 70% of the growth factor content has been delivered. Autologous fibrin matrix retained almost 30% of the amount of the growth factors after 8 days of incubation. The addition of leukocytes to the formula of platelet rich plasma did not increase the concentration of the growth factors, while it drastically increased the presence of pro-inflammatory IL-1β. Further studies employing an in vitro inflammatory model would be interesting to study the difference in growth factors and pro-inflammatory cytokines between leukocyte-free and leukocyte-rich platelet rich plasma. Copyright © 2013 Elsevier GmbH. All rights reserved.

[Analysis of false-positive reaction for bacterial detection of blood samples with the automated microbial detection system BacT/ALERT 3D].

PubMed

Zhu, Li-Wei; Yang, Xue-Mei; Xu, Xiao-Qin; Xu, Jian; Lu, Huang-Jun; Yan, Li-Xing

2008-10-01

This study was aimed to analyze the results of false positive reaction in bacterial detection of blood samples with BacT/ALERT 3D system, to evaluate the specificity of this system, and to decrease the false positive reaction. Each reaction flasks in past five years were processed for bacteria isolation and identification. When the initial cultures were positive, the remaining samples and the corresponding units were recultured if still available. 11395 blood samples were detected. It is worthy of note that the incubator temperature should be stabilized, avoiding fluctuation; when the cultures were alarmed, the reaction flasks showed be kept some hours for further incubation so as to trace a sharply increasing signal to support the judgement of true bacterial growth. The results indicated that 122 samples (1.07%) wee positive at initial culture, out of them 107 samples (88.7%) were found bacterial, and 15 samples (12.3%) were found nothing. The detection curves of positive samples resulted from bacterial growth showed ascent. In conclusion, maintenance of temperature stability and avoidance of temperature fluctuation in incubator could decrease the occurrence of false-positive reaction in detection process. The reaction flasks with positive results at initial culture should be recultured, and whether existence of a sharply ascending logarilhimic growth phase in bacterial growth curve should be further detected, which are helpful to distinguish false-positive reactions from true positive, and thus increase the specificity of the BacT/ALERT system.

Mathematical modelling of thermoregulation processes for premature infants in closed convectively heated incubators.

PubMed

Fraguela, Andrés; Matlalcuatzi, Francisca D; Ramos, Ángel M

2015-02-01

The low-weight newborns and especially the premature infants have difficulty in maintaining their temperature in the range considered to be normal. Several studies revealed the importance of thermal environment and moisture to increase the survival rate of newborns. This work models the process of heat exchange and energy balance in premature newborns during the first hours of life in a closed incubator. In addition, a control problem was proposed and solved in order to maintain thermal stability of premature newborns to increase their rate of survival and weight. For this purpose, we propose an algorithm to control the temperature inside the incubator. It takes into account the measurements of the body temperature of a premature newborn which are recorded continuously. We show that using this model the temperature of a premature newborn inside the incubator can be kept in a thermal stability range. Copyright © 2014. Published by Elsevier Ltd.

Dog Sperm Cryopreservation in Glucose-Fructose or Sucrose Supplemented Glycerol-Free Tris: Effect of Post-Thaw Incubation Time on Gene Expression Related to Apoptosis and Motility.

PubMed

Rahman, M A; Park, S H; Yu, I J

　　OBJECTIVE: The study evaluated the effects of glucose-fructose or sucrose supplementation in glycerol-free Tris (GFT) solution on motility, viability, the level of reactive oxygen species (ROS), as well as the level of apoptosis (BAX and BCL2) and motility (SMCP)-related gene expression of dog spermatozoa. Spermatozoa (5×10 7 sperm/ml) were cryopreserved in GFT containing 86 mM glucose and 86 mM fructose (GF-GFT) or 100 mM sucrose (S-GFT). Progressive motility, viability, ROS (H 2 O 2 ) level and mRNA gene expression of spermatozoa were evaluated 0 h, 3 h or 6 h post-thaw at 24°C. The motility of spermatozoa cryopreserved in GF-GFT was increased throughout the post-thaw incubation time. The motility of spermatozoa cryopreserved in S-GFT was increased at 3 h of post-thaw incubation. The sperm ROS level in the GF-GFT group was inconsistent during the post-thaw incubation time; however, the ROS level in the S-GFT group was gradually increased with progression of the post-thaw incubation period. The post-thaw incubation had no substantial effect on the mRNA expression of the BAX, BCL2, and SMCP genes of spermatozoa in both the GF-GFT and S-GFT groups. The supplementation of glucose and fructose improves progressive sperm motility during 6 h of post-thaw incubation while maintaining similar sperm viability. The addition of GF to GFT for cryopreservation and post-thaw incubation would yield more functional spermatozoa for future assisted reproduction practices.

EFFECT OF VARYING INCUBATION PERIODS ON CYTOTOXICITY AND VIRUCIDAL ACTIVITIES OF Justicia gendarussa Burm.f. LEAF EXTRACT ON HIV-INFECTED MOLT-4 CELLS.

PubMed

Widiyanti, Prihartini; Prajogo, Bambang; Widodo, Agustinus

2018-01-01

Justicia gendarussa Burm.f. has an anti-HIV activity. This study was conducted to evaluate the effects of incubation periods on the cytotoxicity and virucidal activities of the J. gendarussa leaves extract on MOLT-4 cells. The cytotoxicity assay was evaluated by using the WST-1 test with incubation periods of 3 days and 5 days. The virucidal activity test was determined by measuring the inhibitory activities on the syncytium formation. The cytotoxicity assay showed the value of CC 50 on MOLT-4 cell culture with the test material of 70% ethanol extract of J. gendarussa leaves as much as 3928.620 µg /mL and 3176.581 µg /mL (incubation day 3 and day 5, respectively); fractionated-70% ethanol extract = 81782.428 µg /mL and 12175.870 µg/mL; and water extract = 16372.689 µg/mL and 2946.117 µg/mL. The test results of the virucidal activities (inhibit ≥ 90% the formation of syncytium) of 70% ethanol extract of J. gendarussa leaves is at a concentration 250 µg/mL, 500 µg/mL and 1000 µg/mL (3-day incubation) and 250 µg/mL (5-day incubation); and fractionated-70% ethanol extract at a concentration 250 µg /mL, 500 µg/mL and 1000 µg/mL (3-day incubation) and 1000 µg/mL (5-day incubation). 70% ethanol extract, fractionated-70% ethanol extract, and water extract of J. gendarussa leaves were relatively nontoxic toward MOLT-4 cells, and fractionated-70% ethanol extract had better potentials in virucidal activities.

Comparative clinical evaluation of a prototype non-electric transport incubator and an electrical infant incubator in a neonatal unit.

PubMed

Khodadadeh, Y; Nili, F; Nayeri, F; Wickramasinghe, Y

2001-09-01

A new non-electric transport incubator has been developed for transferring babies between health facilities in developing countries. The temperature performance of this prototype was compared with a commercial electric incubator. The warm-up time for the prototype was 51.8 min, compared with 48.1 min for the electric incubator. Forty-five non-distressed premature babies, aged 24-72 h, with a gestational age of less than 37 weeks, were continuously evaluated for a 2 h period. Twenty-five babies, with a mean weight of 2073 g (range 1500-2500 g), were studied in the prototype, and 20 babies, with a mean weight of 2076g (range 1550-2500 g), were studied in the electrical incubator. The rectal and abdominal skin temperature, heart rate, oxygen saturation and respiratory rate of the babies were recorded. The temperature, oxygen and humidity level of the canopy and the room temperature were also measured. The SaO2, heart rate and respiratory rate were within the normal range (in the prototype: 96.5%, 130.5 beats min(-1) and 43 breaths min(-1), respectively; and, in the electric incubator: 96.5%, 128.5 beats min(-1) and 40 breaths min(-1), respectively). No evidence of carbon dioxide narcosis, hypoxia, acidosis or adverse thermoregulatory behaviour were observed in the two groups. The mean rectal temperature for both groups was within the range 36.5 degrees C-37.5 degrees C. There was no significant difference between the measurements of the two groups. The level of oxygen inside the canopy was 21%, and no decrease was observed. The new nonelectric transport incubator confirmed its safety and efficiency in providing a warm environment for non-distressed premature babies over a 2 h period.

Correlation analysis for the incubation period of prion disease.

PubMed

Bae, Se-Eun; Jung, Sunghoon; Kim, Ha-Yeon; Son, Hyeon S

2012-07-01

Previous studies have shown that genetic quantitative trait loci (QTL), strain barriers, inoculation dose and inoculation method modulate the incubation period of prion diseases. We examined the relationship between a diverse set of physical, genetic and immunological characteristics and the incubation period of prion disease using correlation analyses. We found that incubation period was highly correlated with brain weight. In addition, mean corpuscular volume and cell size were strongly correlated with incubation period, indicating that the physical magnitude of prion-infected organs or individual cells may be important in determining the incubation period. Given the same prion inoculation dose, animals with a lower brain weight, mean corpuscular volume or cell size may experience more virulent disease, as the effective concentration of abnormal prion, which might regulate the attachment rate of prions to aggregates, is increased with smaller capacity of brains and cells. This is partly consistent with previous theoretical modeling. The strong correlations between incubation period and physical properties of the brain and cells in this study suggest that the mechanism underlying prion disease pathology may be physical, indicating that the incubation process is governed by simple chemical stoichiometry.

Comparison of the egg flotation and egg candling techniques for estimating incubation day of Canada Goose nests

USGS Publications Warehouse

Reiter, M.E.; Andersen, D.E.

2008-01-01

Both egg flotation and egg candling have been used to estimate incubation day (often termed nest age) in nesting birds, but little is known about the relative accuracy of these two techniques. We used both egg flotation and egg candling to estimate incubation day for Canada Geese (Branta canadensis interior) nesting near Cape Churchill, Manitoba, from 2000 to 2007. We modeled variation in the difference between estimates of incubation day using each technique as a function of true incubation day, as well as, variation in error rates with each technique as a function of the true incubation day. We also evaluated the effect of error in the estimated incubation day on estimates of daily survival rate (DSR) and nest success using simulations. The mean difference between concurrent estimates of incubation day based on egg flotation minus egg candling at the same nest was 0.85 ?? 0.06 (SE) days. The positive difference in favor of egg flotation and the magnitude of the difference in estimates of incubation day did not vary as a function of true incubation day. Overall, both egg flotation and egg candling overestimated incubation day early in incubation and underestimated incubation day later in incubation. The average difference between true hatch date and estimated hatch date did not differ from zero (days) for egg flotation, but egg candling overestimated true hatch date by about 1 d (true - estimated; days). Our simulations suggested that error associated with estimating the incubation day of nests and subsequently exposure days using either egg candling or egg flotation would have minimal effects on estimates of DSR and nest success. Although egg flotation was slightly less biased, both methods provided comparable and accurate estimates of incubation day and subsequent estimates of hatch date and nest success throughout the entire incubation period. ?? 2008 Association of Field Ornithologists.

Biocatalytic Synthesis of the Rare Sugar Kojibiose: Process Scale-Up and Application Testing.

PubMed

Beerens, Koen; De Winter, Karel; Van de Walle, Davy; Grootaert, Charlotte; Kamiloglu, Senem; Miclotte, Lisa; Van de Wiele, Tom; Van Camp, John; Dewettinck, Koen; Desmet, Tom

2017-07-26

Cost-efficient (bio)chemical production processes are essential to evaluate the commercial and industrial applications of promising carbohydrates and also are essential to ensure economically viable production processes. Here, the synthesis of the naturally occurring disaccharide kojibiose (2-O-α-d-glucopyranosyl-d-glucopyranoside) was evaluated using different Bifidobacterium adolescentis sucrose phosphorylase variants. Variant L341I_Q345S was found to efficiently synthesize kojibiose while remaining fully active after 1 week of incubation at 55 °C. Process optimization allowed kojibiose production at the kilogram scale, and simple but efficient downstream processing, using a yeast treatment and crystallization, resulted in more than 3 kg of highly pure crystalline kojibiose (99.8%). These amounts allowed a deeper characterization of its potential in food applications. It was found to have possible beneficial health effects, including delayed glucose release and potential to trigger SCFA production. Finally, we compared the bulk functionality of highly pure kojibiose to that of sucrose, hereby mapping its potential as a new sweetener in confectionery products.

Are We Educating Only Half of the Brain?

ERIC Educational Resources Information Center

Gowan, J. C., Ed.

1979-01-01

The author cites experts on the nature of right hemisphere imagery, the vehicle through which incubation (the second component of creativity) produces creativity, and explores conditions under which imagery occurs. A summarization of the incubation process is provided. (PHR)

Use of serum biochemistry to evaluate nutritional status and health of incubating common eiders (Somateria mollissima) in Finland

USGS Publications Warehouse

Hollmen, T.; Franson, J.C.; Hario, Martti; Sankari, S.; Kilpi, Mikael; Lindstrom, K.

2001-01-01

During 1997–1999, we collected serum samples from 156 common eider (Somateria mollissima) females incubating eggs in the Finnish archipelago of the Baltic Sea. We used serum chemistry profiles to evaluate metabolic changes in eiders during incubation and to compare the health and nutritional status of birds nesting at a breeding area where the eider population has declined by over 50% during the past decade, with birds nesting at two areas with stable populations. Several changes in serum chemistries were observed during incubation, including (1) decreases in serum glucose, total protein, albumin, β‐globulin, and γ‐globulin concentrations and (2) increases in serum uric acid, creatine kinase, and β‐hydroxybutyrate concentrations. However, these changes were not consistent throughout the 3‐yr period, suggesting differences among years in the rate of carbohydrate, lipid, and protein utilization during incubation. The mean serum concentrations of free fatty acids, glycerol, and albumin were lowest and the serum α‐ and γ‐globulin levels were highest in the area where the eider population has declined, suggesting a role for nutrition and diseases in the population dynamics of Baltic eiders.

Human renal adipose tissue induces the invasion and progression of renal cell carcinoma.

PubMed

Campo-Verde-Arbocco, Fiorella; López-Laur, José D; Romeo, Leonardo R; Giorlando, Noelia; Bruna, Flavia A; Contador, David E; López-Fontana, Gastón; Santiano, Flavia E; Sasso, Corina V; Zyla, Leila E; López-Fontana, Constanza M; Calvo, Juan C; Carón, Rubén W; Creydt, Virginia Pistone

2017-11-07

We evaluated the effects of conditioned media (CMs) of human adipose tissue from renal cell carcinoma located near the tumor (hRATnT) or farther away from the tumor (hRATfT), on proliferation, adhesion and migration of tumor (786-O and ACHN) and non-tumor (HK-2) human renal epithelial cell lines. Human adipose tissues were obtained from patients with renal cell carcinoma (RCC) and CMs from hRATnT and hRATfT incubation. Proliferation, adhesion and migration were quantified in 786-O, ACHN and HK-2 cell lines incubated with hRATnT-, hRATfT- or control-CMs. We evaluated versican, adiponectin and leptin expression in CMs from hRATnT and hRATfT. We evaluated AdipoR1/2, ObR, pERK, pAkt y pPI3K expression on cell lines incubated with CMs. No differences in proliferation of cell lines was found after 24 h of treatment with CMs. All cell lines showed a significant decrease in cell adhesion and increase in cell migration after incubation with hRATnT-CMs vs. hRATfT- or control-CMs. hRATnT-CMs showed increased levels of versican and leptin, compared to hRATfT-CMs. AdipoR2 in 786-O and ACHN cells decreased significantly after incubation with hRATfT- and hRATnT-CMs vs. control-CMs. We observed a decrease in the expression of pAkt in HK-2, 786-O and ACHN incubated with hRATnT-CMs. This result could partially explain the observed changes in migration and cell adhesion. We conclude that hRATnT released factors, such as leptin and versican, could enhance the invasive potential of renal epithelial cell lines and could modulate the progression of the disease.

Paradoxical toxicity of cardioplegic compounds on ischemic cardiomyocyte using optimal design strategy.

PubMed

Ferrera, René; Michel, Pierre; Ovize, Michel

2005-07-01

The aim of this study was to evaluate the effects of major components of cardioplegic solutions on myocardial tissue submitted to prolonged cold ischemia. Our methodology was based on the simultaneous testing in the same series of experiments of many compounds (19 in number), which were included in the composition of 20 established solutions. All the experiments were performed by a matricial-predefined protocol that allows the evaluation of the protective or toxic effects of each of these 19 compounds. Pig hearts were removed and left ventricular myocardiums were cut into 320 pieces. For each solution tested, 8 pieces of myocardial tissue were incubated at 4 degrees C for 24 hours and 8 other pieces were incubated for 72 hours. At the end of incubation period, tissue injury was assessed by measuring the leakage of myocardial enzymes(glutamic-oxaloacetic transaminase, lactate dehydrogenase, creatine phosphokinase) into the incubation medium. Initially, the effects of each solution were evaluated, and then a mathematical analysis was performed and the effects of each compound deduced. After the 24-hour incubation period, pyruvate (5 mmol/liter), polyethylene glycol (5 mmol/liter), Ala-Gln (20 mmol/liter), and reduced glutathione (3 mmol/liter) showed toxic effects, whereas ethanol (1%) and calcium chloride (2 mmol/liter) seemed to be protective. After 72 hours' incubation, similar data were obtained; dextran 70 (0.57 mmol/liter) was also found to be deleterious. The results revealed surprising myocardial toxicity (enzymatic release) from components included in cardioplegic solutions. Some components would induce metabolic activation during prolonged hypothermic ischemia, which may be inappropriated and which may perhaps exacerbate damages by increasing membrane ruptures. This concept confirms eventual discrepant effects of preservative compounds on cardiomyocyte membrane during deep hypothermia, according to the metabolic state of the cell.

Toward a fuzzy logic control of the infant incubator.

PubMed

Reddy, Narender P; Mathur, Garima; Hariharan, S I

2009-10-01

Premature birth is a world wide problem. Thermo regulation is a major problem in premature infants. Premature infants are often kept in infant incubators providing convective heating. Currently either the incubator air temperature is sensed and used to control the heat flow, or infant's skin temperature is sensed and used in the close loop control. Skin control often leads to large fluctuations in the incubator air temperature. Air control also leads to skin temperature fluctuations. The question remains if both the infant's skin temperature and the incubator air temperature can be simultaneously used in the control. The purpose of the present study was to address this question by developing a fuzzy logic control which incorporates both incubator air temperature and infant's skin temperature to control the heating. The control was evaluated using a lumped parameter mathematical model of infant-incubator system (Simon, B. N., N. P. Reddy, and A. Kantak, J. Biomech. Eng. 116:263-266, 1994). Simulation results confirmed previous experimental results that the on-off skin control could lead to fluctuations in the incubator air temperature, and the air control could lead to too slow rise time in the core temperature. The fuzzy logic provides a smooth control with the desired rise time.

A Caco-2 cell-based quantitative antioxidant activity assay for antioxidants.

PubMed

Wan, Hongxia; Liu, Dong; Yu, Xiangying; Sun, Haiyan; Li, Yan

2015-05-15

A Caco-2 cell-based antioxidant activity (CAA) assay for quantitative evaluation of antioxidants was developed by optimizing seeding density and culture time of Caco-2 cells, incubation time and concentration of fluorescent probe (2',7'-dichlorofluorescin diacetate, DCFH-DA), incubation way and incubation time of antioxidants (pure phytochemicals) and DCFH-DA with cells, and detection time of fluorescence. Results showed that the CAA assay was of good reproducibility and could be used to evaluate the antioxidant activity of antioxidants at the following conditions: seeding density of 5 × 10(4)/well, cell culture time of 24h, co-incubation of 60 μM DCFH-DA and pure phytochemicals with Caco-2 cells for 20 min and fluorescence recorded for 90 min. Additionally, a significant correlation was observed between CAA values and rat plasma ORAC values following the intake of antioxidants for selected pure phytochemicals (R(2) = 0.815, p < 0.01), demonstrating the good biological relevance of CAA assay. Copyright © 2014 Elsevier Ltd. All rights reserved.

Short-term effects of natural and NH4+-enriched chabazite zeolitite amendments to soil microbial biomass

NASA Astrophysics Data System (ADS)

Ferretti, Giacomo; Keiblinger, Katharina Maria; Di Giuseppe, Dario; Faccini, Barbara; Colombani, Nicolò; Zechmeister-Boltenstern, Sophie; Coltorti, Massimo; Mastrocicco, Micòl

2017-04-01

Natural zeolite-bearing rocks (zeolitites) are known to be a suitable material for agricultural purposes by improving soil physicochemical properties and nitrogen use efficiency (NUE). However, little is known about their effects on soil microbial biomass. Aim of this work is to evaluate short-term effects of different chabazite-zeolitite amendments on soil microbial biomass (and activity). To this purpose a silty-clay agricultural soil was amended in three different ways, by the addition of 5 and 15 wt% of natural chabazite zeolitites (NZ) and 10 wt% of NH4+-enriched chabazite zeolitites (CZ). Soil pH, water content, dissolved organic carbon (C), total dissolved N, NH4+, NO3-, NO2-, microbial biomass C and N and ergosterol were periodically measured over a time course of 16 days in a laboratory incubation experiment. In order to verify the immobilization of N derived from CZ into microbial biomass, the δ15N signature of microorganisms was evaluated by the Extraction-Fumigation-Extraction method followed by EA-IRMS analysis. This latter investigation was possible because zeolitites were enriched with NH4+ derived from pig-slurry, which have a very high 15N natural abundance that allow to trace microbial incorporation. Soil amended with 5 wt% of NZ showed increased ergosterol content as well as microbial C/N ratio starting from day 9 of incubation, suggesting that fungal biomass was probably favored, although the same behavior was not found in the soil amended with 15 wt% of the same material. On the other hand, the NH4+-enriched CZ showed strong interactions with soil microbial biomass N. Isotopic measurements supported microbial assimilation of the N introduced with CZ since the second day of incubation. The high dissolved organic C and microbial biomass N suggested an increase of mineralization and immobilization processes. In addition, in CZ amended soil, microbial biomass N was related to NO3- production over time and inversely related to NH4+, suggesting high nitrification processes especially from day 7 of incubation. Low microbial C/N ratio support bacterial prevalence in the soil amended with CZ for N-assimilation and ammonia oxidation. This confirm that CZ is an efficient soil amendment providing an immediately available N pool to soil microbial biomass.

Effect of genistein added to bull semen after thawing on pronuclear and sperm quality.

PubMed

Silvestre, M A; Vicente-Fiel, S; Raga, E; Salvador, I; Soler, C; Yániz, J L

2015-12-01

The aim of this research was to study the effect of different genistein treatments on bull sperm after thawing on pronuclear formation after in vitro fertilization (IVF) and on different sperm quality variables. Three experiments were performed. In Experiment 1, three treatments (Control, sperm incubation for 1h at 37 °C with or without genistein) and two sperm concentrations during IVF (1 or 3 × 10(6)sperm/mL) were evaluated to study the influence of genistein on pronuclear formation (PNF). Sperm incubation for 1h before IVF reduced PNF regardless of sperm concentration. However, after sperm incubation and with 3 × 10(6)sperm/mL in IVF, the genistein treatment group had greater fertilization rates than the untreated group. In Experiment 2, six treatments plus the control group were performed to study the effect of genistein (presence or not) and incubation conditions (30 min at 37 °C, 1h at 27 °C or at 37 °C) on PNF using 3 × 10(6)sperm/mL for IVF. When incubation time was reduced to 30 min, PNF rate from the genistein treatment group was no different from either the control group or in the group in which incubation occurred for 1h at 27 °C. In Experiment 3, the effect of several genistein treatments (control; genistein treatment for 30 min of incubation at 37 °C; genistein treatment for 1h of incubation at 27 °C) on sperm motility, viability and DNA fragmentation were evaluated. Genistein did not improve sperm motility and, depending on the experimental group or time, it either reduced or had no effect on sperm motility. Genistein treatment did not improve sperm viability after 5h of incubation. However, genistein treatment for 1h at 27 °C decreased sperm DNA fragmentation compared with the control group after 5h of sperm incubation. In conclusion, the treatment of bull sperm with genistein for 1h at 27 °C could decrease sperm DNA fragmentation, although PNF rate after IVF and sperm motility were reduced. Copyright © 2015. Published by Elsevier B.V.

Biosynthesis of gold nanoparticles using Capsicum annuum var. grossum pulp extract and its catalytic activity

NASA Astrophysics Data System (ADS)

Yuan, Chun-Gang; Huo, Can; Yu, Shuixin; Gui, Bing

2017-01-01

Biological synthesis approach has been regarded as a green, eco-friendly and cost effective method for nanoparticles preparation without any toxic solvents and hazardous bi-products during the process. This present study reported a facile and rapid biosynthesis method for gold nanoparticles (GNPs) from Capsicum annuum var. grossum pulp extract in a single-pot process. The aqueous pulp extract was used as biotic reducing agent for gold nanoparticle growing. Various shapes (triangle, hexagonal, and quasi-spherical shapes) were observed within range of 6-37 nm. The UV-Vis spectra showed surface plasmon resonance (SPR) peak for the formed GNPs at 560 nm after 10 min incubation at room temperature. The possible influences of extract amount, gold ion concentration, incubation time, reaction temperature and solution pH were evaluated to obtain the optimized synthesis conditions. The effects of the experimental factors on NPs synthesis process were also discussed. The produced gold nanoparticles were characterized by transform electron microscopy (TEM), X-ray diffraction (XRD), energy dispersive X-ray (EDS) and Fourier Transform infrared spectroscopy (FTIR). The results demonstrated that the as-obtained GNPs were well dispersed and stable with good catalytic activity. Biomolecules in the aqueous extract were responsible for the capping and stabilization of GNPs.

Sperm preparation through Sephadex™ filtration improves in vitro fertilization rate of buffalo oocytes.

PubMed

Husna, A U; Azam, A; Qadeer, S; Awan, M A; Nasreen, S; Shahzad, Q; Fouladi-Nashta, A; Khalid, M; Akhter, S

2018-04-01

Routinely, swim-up method is used to separate high-quality sperm; however, long processing time and close cell-to-cell contact during the centrifugation step are inevitable elements of oxidative stress to sperm. The objective was to evaluate Sephadex ™ and glass wool filtration to separate motile, intact and viable sperm for in vitro fertilization in buffalo. The cumulus-oocyte complexes (COCs) were collected from ovaries of slaughtered buffaloes by aspiration and matured for 24 hr in CO 2 incubator at 38.5°C and 5% CO 2 . Matured COCs were rinsed twice in fertilization TALP and placed in the pre-warmed fertilization medium without sperm. Cryopreserved buffalo semen was thawed at 37°C for 30 s and processed through Sephadex ™ , glass wool filtration and swim-up (control). Total and motile sperm recovery rates were assessed, resuspended in fertilization TALP and incubated for 15-20 min in CO 2 incubator. Samples prepared by each method were divided into two aliquots: one aliquot was studied for sperm quality (progressive motility, membrane integrity, viability, liveability), while the other was subjected to co-incubation with sets of 10-15 in vitro matured oocytes. Data on sperm quality were analysed by ANOVA, while in vitro fertilizing rates were compared by chi-squared test using SPSS-20. Least significant difference (LSD) test was used to compare treatment means. Glass wool filtration yielded higher total and motile sperm recovery rate, while Sephadex ™ filtration improved (p < .05) sperm quality (progressive motility, membrane integrity, viability, liveability). Sperm preparation through Sephadex filtration yielded higher in vitro fertilization rate in terms of cleavage rate compared to glass wool filtration and swim-up (control). In conclusion, cryopreserved Nili-Ravi buffalo sperm selected through Sephadex filtration showed improved quality and yielded better fertilization rates (cleavage rate) of in vitro matured/fertilized oocytes. Sephadex filtration could be a promising technique for use in in vitro fertilization in buffalo. © 2017 Blackwell Verlag GmbH.

Caffeine strongly improves motility parameters of turkey spermatozoa with no effect on cell viability.

PubMed

Slanina, Tomáš; Miškeje, Michal; Tirpák, Filip; Błaszczyk, Martyna; Formicki, Grzegorz; Massányi, Peter

2018-03-01

The purpose of this study was to evaluate the impact of caffeine on turkey spermatozoa during in vitro incubation. Experimental samples were prepared by diluting the raw semen with nine different concentrations of caffeine - from 0.078125 mg/mL to 10 mg/mL. The individual motility parameters were evaluated by the Computer Assisted Semen Analyser (CASA) system, and the viability of spermatozoa was evaluated using eosin-nigrosin staining. Selected parameters were recorded at six time periods: 0, 1, 2, 3, 4 and 5 h at 5 °C and 41 °C. A significantly higher motility and progressive motility of spermatozoa (P < 0.01 and P < 0.001, respectively) was detected in the samples containing caffeine ranging from 0.15625 to 7.5 mg/mL as compared to the control sample at 5 °C. At an incubation temperature of 41 °C the positive effect of caffeine on motility parameters was observed only at the beginning of incubation (at times 0 and 1). The tested caffeine concentrations showed no significant effect on the viability of turkey spermatozoa at any time period of incubation. A higher percentage of dead spermatozoa was observed for incubation at 41 °C (from 5.96% to 11.1%) in comparison to 5 °C (from 1.62% to 5.79%). The results suggest that caffeine can be used as a suitable component of turkey semen extenders and has the potential to improve fertility.

Knowledge service decision making in business incubators based on the supernetwork model

NASA Astrophysics Data System (ADS)

Zhao, Liming; Zhang, Haihong; Wu, Wenqing

2017-08-01

As valuable resources for incubating firms, knowledge resources have received gradually increasing attention from all types of business incubators, and business incubators use a variety of knowledge services to stimulate rapid growth in incubating firms. Based on previous research, we generalize the knowledge transfer and knowledge networking services of two main forms of knowledge services and further divide knowledge transfer services into knowledge depth services and knowledge breadth services. Then, we construct the business incubators' knowledge supernetwork model, describe the evolution mechanism among heterogeneous agents and utilize a simulation to explore the performance variance of different business incubators' knowledge services. The simulation results show that knowledge stock increases faster when business incubators are able to provide knowledge services to more incubating firms and that the degree of discrepancy in the knowledge stock increases during the process of knowledge growth. Further, knowledge transfer services lead to greater differences in the knowledge structure, while knowledge networking services lead to smaller differences. Regarding the two types of knowledge transfer services, knowledge depth services are more conducive to knowledge growth than knowledge breadth services, but knowledge depth services lead to greater gaps in knowledge stocks and greater differences in knowledge structures. Overall, it is optimal for business incubators to select a single knowledge service or portfolio strategy based on the amount of time and energy expended on the two types of knowledge services.

Resistance of soybean genotypes to Sclerotinia sclerotiorum isolates in different incubation environments

USDA-ARS?s Scientific Manuscript database

Sclerotinia sclerotiorum is an important soybean pathogen. The objectives of this study were to evaluate levels of resistance of soybean genotypes to the fungus, and to determine the effects of different incubation environments on host resistance and pathogen aggression. Two experiments were conduct...

External Service Providers to the National Security Technology Incubator: Formalization of Relationships

DOE Office of Scientific and Technical Information (OSTI.GOV)

None

2008-04-30

This report documents the formalization of relationships with external service providers in the development of the National Security Technology Incubator (NSTI). The technology incubator is being developed as part of the National Security Preparedness Project (NSPP), funded by a Department of Energy (DOE)/National Nuclear Security Administration (NNSA) grant. This report summarizes the process in developing and formalizing relationships with those service providers and includes a sample letter of cooperation executed with each provider.

Effects of novel nitrification and urease inhibitors (DCD/TZ and 2-NPT) on N2O emissions from surface applied urea: An incubation study

NASA Astrophysics Data System (ADS)

Ni, Kang; Kage, Henning; Pacholski, Andreas

2018-02-01

A 41-day incubation trial was conducted to test the single and combined effects of the novel urease (N-(2-Nitrophenyl) phosphoric triamide, 2-NPT) and nitrification inhibitors (mixture of dicyandiamide and 1H-1,2,4-triazole, DCD/TZ) on N2O emissions and underlying soil processes from a North German sandy loam soil. The effects of treatment on N2O emission were determined using static closed chamber incubation and detected using a photo-acoustic gas monitor. The emission processes were strongly related to soil mineral N and pH dynamics, obtained from destructive sampling of replicate incubation chambers. The combined use of urease and nitrification inhibitors slightly increased the reduction of N2O compared with single use of the nitrification inhibitor (69% vs. 61%). The small amount of soil used in the incubation and the depletion of labile carbon by air drying and pre-incubation caused very low initial N2O emissions, and glucose addition significantly stimulated N2O emission by supplying labile carbon. The urease inhibitor significantly reduced simultaneously determined qualitative NH3 emissions in either urea alone (90%) or urea plus nitrification inhibitor treatment (82%). These results highlighted the potential of the combined use of urease and nitrification inhibitors with urea application to mitigate soil NH3 and N2O emissions.

Parkia pendula lectin as histochemistry marker for meningothelial tumour.

PubMed

Beltrão, E I C; Medeiros, P L; Rodrigues, O G; Figueredo-Silva, J; Valença, M M; Coelho, L C B B; Carvalho, L B

2003-01-01

Lectins have been intensively used in histochemical techniques for cell surface characterization. These proteins are involved in several biological processes and their use as histochemical markers have been evaluated since they can indicate differences in cell surfaces. Parkia pendula lectin (PpeL) was evaluated as histochemical marker for meningothelial meningioma biopsies. Tissue slices were incubated with PpeL conjugated to horseradish peroxidase (PpeL-HRP) and Concanavalin A-HRP (ConA-HPR) and the binding visualized with diaminobenzidine and hydrogen peroxide. The lectin-tissue binding was inhibited with D-glucose. PpeL showed to be a useful tool for the characterization of meningothelial tumour and clinico-pathological diagnosis.

Correlation analysis for the incubation period of prion disease

PubMed Central

Bae, Se-Eun; Jung, Sunghoon; Kim, Ha-Yeon; Son, Hyeon S.

2012-01-01

Previous studies have shown that genetic quantitative trait loci (QTL), strain barriers, inoculation dose and inoculation method modulate the incubation period of prion diseases. We examined the relationship between a diverse set of physical, genetic and immunological characteristics and the incubation period of prion disease using correlation analyses. We found that incubation period was highly correlated with brain weight. In addition, mean corpuscular volume and cell size were strongly correlated with incubation period, indicating that the physical magnitude of prion-infected organs or individual cells may be important in determining the incubation period. Given the same prion inoculation dose, animals with a lower brain weight, mean corpuscular volume or cell size may experience more virulent disease, as the effective concentration of abnormal prion, which might regulate the attachment rate of prions to aggregates, is increased with smaller capacity of brains and cells. This is partly consistent with previous theoretical modeling. The strong correlations between incubation period and physical properties of the brain and cells in this study suggest that the mechanism underlying prion disease pathology may be physical, indicating that the incubation process is governed by simple chemical stoichiometry. PMID:22561168

Uterosome-like vesicles prompt human sperm fertilizing capability.

PubMed

Franchi, A; Cubilla, M; Guidobaldi, H A; Bravo, A A; Giojalas, L C

2016-12-01

Does the rapid transit through the uterine environment modulate the sperm physiological state? The uterosome-like vesicles (ULVs) secreted by endometrial epithelial cells (EECs) in vitro are able to fuse with human spermatozoa, prompting their fertilizing capacity. Early studies suggest that sperm capacitation begins in the uterus and ends in the oviduct, and that a synergistic effect of both female organs may accelerate this process. Although it has been reported that co-incubation of human spermatozoa with endometrial cell-conditioned medium (CM) stimulates sperm capacitation, the mechanism mediating this communication is unknown. Human ULVs secreted by EECs were characterized and their effect on human sperm physiology was analysed. Spermatozoa were incubated with EEC-derived CM or ULV, after which sperm capacitation was evaluated at different time points. In addition, the interaction of spermatozoa with ULV was analysed. ULVs were isolated by ultracentrifugation and identified using electron microscopy and Western blotting to assess the presence of specific protein markers. Following seminal plasma removal, human spermatozoa were incubated CM or ULV, after which sperm capacitation was evaluated as the ability of the sperm to undergo the induced acrosome reaction and the level of protein tyrosine phosphorylation (PY) determined by Western blot and immunocytochemistry. The interaction of spermatozoa with labelled ULV was analysed by fluorescence microscopy. In all cases, at least three biological replicates from different sperm donors were performed for each set of experiments. Significant differences between mean values were determined by one-way ANOVA followed by Tukey's post hoc test. Differences between treatments were considered statistically significant at P ≤ 0.05. The level of capacitated spermatozoa and those recruited by chemotaxis increased 3- to 4-fold when spermatozoa were incubated in the presence of CM for 4 h. Even a 15 min incubation of spermatozoa with CM was also enough to increase the level of capacitated cells 3- to 4-fold (P < 0.05). Furthermore, a short co-incubation of spermatozoa with ULV stimulates sperm capacitation, as determined by the increase in the level of induced acrosome reaction and the induction of PY. In addition, after the co-incubation of spermatozoa with fluorescent labelled ULV, the sperm cells acquired the fluorescent staining which indicates that ULV might be transferred to the sperm surface by a fusion mechanism. This is an in vitro study performed with human biological material, spermatozoa and endometrial derived cells; the latter being a cell line originally isolated from a uterine adenocarcinoma. The capability of spermatozoa to briefly interact with ULVs supports the hypothesis that any step of sperm transport may have physiological consequences, despite the interaction lasting for only a limited period of time. This way of communication of spermatozoa with cell products of uterine origin opens new frontiers of investigation (e.g. the signalling molecules involved), shedding light on the sperm processes that prepare the male gamete for fertilization, which might have implications for human infertility treatment. N/A. The project was financially supported by SECyT-UNC. The authors declare no conflict of interest. © The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Incubation stage and polychlorinated biphenyl (PCB) congener patterns in an altricial and precocial bird species

USGS Publications Warehouse

Custer, Christine M.; Custer, Thomas W.; Thyen, Stefan; Becker, Peter H.

2014-01-01

The composition of polychlorinated biphenyl (PCB) congeners was compared between non-incubated and embryonated eggs of tree swallows (Tachycineta bicolor) and little terns (Sterna albifrons) to determine if measurable changes in PCB congeners occurred during the embryonic period. There was no indication of changes in PCB congener patterns over the incubation period in tree swallows in 1999 and 2000 at a site with very high PCB exposure or a site with more modest PCB exposure. Additionally, congeners known to be either quickly metabolized or conserved based on experimental studies did not generally respond as predicted. Similarly, PCB congener patterns in eggs of little terns from Bottsand, Schleswig-Holstein, Germany, did not differ between non-incubated and embryonated eggs. The results from both species suggest that the stage of incubation is not an important consideration when evaluating PCB congener patterns; comparisons and assessments can be made with eggs collected at all stages of incubation.

Incubation stage and polychlorinated biphenyl (PCB) congener patterns in an altricial and precocial bird species.

PubMed

Custer, Christine M; Custer, Thomas W; Thyen, Stefan; Becker, Peter H

2014-12-01

The composition of polychlorinated biphenyl (PCB) congeners was compared between non-incubated and embryonated eggs of tree swallows (Tachycineta bicolor) and little terns (Sterna albifrons) to determine if measurable changes in PCB congeners occurred during the embryonic period. There was no indication of changes in PCB congener patterns over the incubation period in tree swallows in 1999 and 2000 at a site with very high PCB exposure or a site with more modest PCB exposure. Additionally, congeners known to be either quickly metabolized or conserved based on experimental studies did not generally respond as predicted. Similarly, PCB congener patterns in eggs of little terns from Bottsand, Schleswig-Holstein, Germany, did not differ between non-incubated and embryonated eggs. The results from both species suggest that the stage of incubation is not an important consideration when evaluating PCB congener patterns; comparisons and assessments can be made with eggs collected at all stages of incubation. Published by Elsevier Ltd.

Proximal tubule hydrogen ion transport processes in diuretic-induced metabolic alkalosis.

PubMed

Blumenthal, S S; Ware, R A; Kleinman, J G

1985-07-01

Transport systems involved in proximal tubule HCO-3 reabsorption were examined in disaggregated renal cortical tubules from rabbits with metabolic alkalosis. The acid-base disorder was induced by first treating the animals with furosemide, and then maintaining them on low Cl--high HCO-3 diets. On this regimen, the rabbits had increases in blood pH and total CO2 values and decreases in serum K+ concentrations. Urine Cl- concentrations were less than 15 mEq/L in all cases. Na+-H+ exchange was evaluated by incubating tubules in rotenone in an Na+-free medium to deplete them of Na+ and adenosine triphosphate. Then the tubules were resuspended in media containing 65 or 12.5 mEq/L Na+ at either pH 7.1 or pH 7.6. The rise in cell pH estimated by dimethadione distribution was taken as a measure of Na+-H+ exchanger activity. At the high incubation pH, Na+-H+ exchanger activity appeared to be the same in tubules taken from alkalotic rabbits compared with those prepared from normal rabbits. At the low incubation pH, the activity of this transport system appeared to be depressed by 40% to 50% in alkalosis, with kinetics that suggested a decreased Vmax for the exchanger. Na+-independent H+ transport, presumably reflecting activity of an H+-adenosine triphosphatase, was evaluated by preincubating tubules in a Na+-free medium in the presence of ouabain, and then sequentially exposing them to and removing them from a solution containing 20 mmol/L NH4Cl.(ABSTRACT TRUNCATED AT 250 WORDS)

Incubation environment impacts the social cognition of adult lizards.

PubMed

Siviter, Harry; Deeming, D Charles; van Giezen, M F T; Wilkinson, Anna

2017-11-01

Recent work exploring the relationship between early environmental conditions and cognition has shown that incubation environment can influence both brain anatomy and performance in simple operant tasks in young lizards. It is currently unknown how it impacts other, potentially more sophisticated, cognitive processes. Social-cognitive abilities, such as gaze following and social learning, are thought to be highly adaptive as they provide a short-cut to acquiring new information. Here, we investigated whether egg incubation temperature influenced two aspects of social cognition, gaze following and social learning in adult reptiles ( Pogona vitticeps ). Incubation temperature did not influence the gaze following ability of the bearded dragons; however, lizards incubated at colder temperatures were quicker at learning a social task and faster at completing that task. These results are the first to show that egg incubation temperature influences the social cognitive abilities of an oviparous reptile species and that it does so differentially depending on the task. Further, the results show that the effect of incubation environment was not ephemeral but lasted long into adulthood. It could thus have potential long-term effects on fitness.

Incubation environment impacts the social cognition of adult lizards

PubMed Central

van Giezen, M. F. T.

2017-01-01

Recent work exploring the relationship between early environmental conditions and cognition has shown that incubation environment can influence both brain anatomy and performance in simple operant tasks in young lizards. It is currently unknown how it impacts other, potentially more sophisticated, cognitive processes. Social-cognitive abilities, such as gaze following and social learning, are thought to be highly adaptive as they provide a short-cut to acquiring new information. Here, we investigated whether egg incubation temperature influenced two aspects of social cognition, gaze following and social learning in adult reptiles (Pogona vitticeps). Incubation temperature did not influence the gaze following ability of the bearded dragons; however, lizards incubated at colder temperatures were quicker at learning a social task and faster at completing that task. These results are the first to show that egg incubation temperature influences the social cognitive abilities of an oviparous reptile species and that it does so differentially depending on the task. Further, the results show that the effect of incubation environment was not ephemeral but lasted long into adulthood. It could thus have potential long-term effects on fitness. PMID:29291066

Estimating Supplies Program: Evaluation Report

DTIC Science & Technology

2002-12-24

Inhalation, Non-vaccinated1, Incubating, Asymptomatic 352 Anthrax, Inhalation, Non-vaccinated, Prodromal 353 Anthrax, Inhalation, Non-vaccinated, Acute...B-11 PC Code PC Description 354 Anthrax, Inhalation, Vaccinated, Asymptomatic 355 Anthrax, Inhalation, Vaccinated, Prodromal 356...Anthrax, Inhalation, Vaccinated, Acute 357 Plague, Inhalation, Incubating, Asymptomatic 358 Plague, Inhalation, Acute 359 Plague Meningitis 360

Utilizing pretreatment and fungal incubation to enhance the nutritional value of canola meal

USDA-ARS?s Scientific Manuscript database

The objective of this study was to determine the optimal pretreatment and fungal strain to reduce glucosinolates (GLS), fiber, and residual sugars while increasing the nutritional value of canola meal. Submerged incubation conditions were used to evaluate four pretreatment methods (extrusion, hot wa...

Effect of a photoperiodic green light programme during incubation on embryo development and hatch process.

PubMed

Tong, Q; McGonnell, I M; Demmers, T G M; Roulston, N; Bergoug, H; Romanini, C E; Verhelst, R; Guinebretière, M; Eterradossi, N; Berckmans, D; Exadaktylos, V

2018-04-01

This study was conducted to evaluate the effect of a 12-h light, 12-h dark (12L : 12D) photoperiod of green light during day 1 to day 18 of incubation time, on embryo growth, hormone concentration and the hatch process. In the test group, monochromatic light was provided by a total of 204 green light-emitting diodes (522 nm) mounted in a frame which was placed above the top tray of eggs to give even spread of illumination. No light-dark cycle was used in the control group. Four batches of eggs (n=300/group per batch) from fertile Ross 308 broiler breeders were used in this experiment. The beak length and crown-rump length of embryos incubated under green light were significantly longer than that of control embryos at day 10 and day 12, respectively (P<0.01). Furthermore, green light-exposed embryos had a longer third toe length compared with control embryos at day 10, day 14 and day 17 (P=0.02). At group level (n=4 batches), light stimulation had no effect on chick weight and quality at take-off, the initiation of hatch and hatch window. However, the individual hatching time of the light exposure focal chicks (n=33) was 3.4 h earlier (P=0.49) than the control focal chicks (n=36) probably due to the change in melatonin rhythm of the light group. The results of this study indicate that green light accelerates embryo development and alters hatch-related hormones (thyroid and corticosterone), which may result in earlier hatching.

Dynamics of sperm subpopulations based on motility and plasma membrane status in thawed ram spermatozoa incubated under conditions that support in vitro capacitation and fertilisation.

PubMed

García-Álvarez, Olga; Maroto-Morales, Alejandro; Ramón, Manuel; del Olmo, Enrique; Jiménez-Rabadán, Pilar; Fernández-Santos, M Rocio; Anel-López, Luis; Garde, J Julián; Soler, Ana J

2014-06-01

The present study evaluated modifications occurring in thawed ram spermatozoa during incubation in different media that supported in vitro capacitation and fertilisation, and examines how these changes relate to IVF. Thawed sperm samples were incubated under capacitating (Cap) and non-capacitating (non-Cap) conditions for 0, 1 and 2h and used in an IVF test. During incubation, changes related to membrane status and the motility pattern of spermatozoa were assessed, the latter being used to characterise sperm subpopulations. A significantly greater increase (P≤0.05) in the percentage of spermatozoa with higher membrane fluidity was observed in samples incubated with Cap medium from the beginning of incubation. In addition, changes over time in the distribution of the motile subpopulation were particularly evident when spermatozoa were incubated with Cap medium, with a noted increase in spermatozoa classified as 'hyperactivated like', with major changes occurring after 1h incubation. Both characteristics (i.e. membrane fluidity and the percentage of the hyperactivated-like subpopulation) were significantly related with in vitro fertility, and only sperm samples incubated with the Cap medium were capable of fertilising oocytes. These results support the idea that changes in sperm membrane fluidity and motility pattern (i.e. an increase in hyperactivated spermatozoa) are needed for fertilisation to take place.

The incubation period of cholera: a systematic review.

PubMed

Azman, Andrew S; Rudolph, Kara E; Cummings, Derek A T; Lessler, Justin

2013-05-01

Recent large cholera outbreaks highlight the need for improved understanding of the pathogenesis and epidemiology of cholera. The incubation period of cholera has important implications for clinical and public health decision-making, yet statements of the incubation period of cholera are often imprecise. Here we characterize the distribution of cholera's incubation period. We conducted a systematic review of the literature for statements of the incubation period of cholera and data that might aid in its estimation. We extracted individual-level data, parametrically estimated the distribution of toxigenic cholera's incubation period, and evaluated evidence for differences between strains. The incubation period did not differ by a clinically significant margin between strains (except O1 El Tor Ogawa). We estimate the median incubation period of toxigenic cholera to be 1.4 days (95% CI, 1.3-1.6). Five percent of cholera cases will develop symptoms by 0.5 days (95% CI 0.4-0.5), and 95% by 4.4 days (95% CI 3.9-5.0) after infection. We recommend that cholera investigations use a recall period of at least five days to capture relevant exposures; significantly longer than recent risk factor studies from the Haitian epidemic. This characterization of cholera's incubation period can help improve clinical and public health practice and advance epidemiologic research. Copyright © 2012 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

Effect of temperature, anaerobiosis, stirring and salt addition on natural fermentation silage of sardine and sardine wastes in sugarcane molasses.

PubMed

Zahar, M; Benkerroum, N; Guerouali, A; Laraki, Y; El Yakoubi, K

2002-04-01

Conditions for a natural fermentation during ensilage of sardines or their waste in sugarcane molasses (60:40 w/w) were evaluated regarding the effect of temperature (15, 25 and 35 degrees C), anaerobiosis (closed vs. open jars), daily stirring of the mixture, and salt addition to the initial mix at 5% (w/w) level. Successful natural fermentation took place in sardine silages incubated at 25 or 35 degrees C in open jars to reach a pH of 4.4 in about 2 and 1 weeks, respectively. For samples kept at 15 degrees C, the pH decline was very slow and pH did not decrease below 5.5 after one month of incubation. At 25 degrees C, the most favorable conditions for silage of sardine waste in cane molasses, as evidenced by the fastest decline in pH to a stable value of about 4.4, were achieved in closed jars and with daily stirring of the mix. The pH 4.4 was reached in one week with an advance of at least 3 days compared to the other conditions (open jars and closed jars without daily stirring). Addition of salt at 5% (w/w) in the mix before incubation inhibited the fermentation process.

Potential use of cowpea (Vigna unguiculata (L.) Walp.) stover treated with white-rot fungi as rabbit feed.

PubMed

Andrade, Ederson; Pinheiro, Victor; Gonçalves, Alexandre; Cone, John W; Marques, Guilhermina; Silva, Valéria; Ferreira, Luis; Rodrigues, Miguel

2017-10-01

Lignin inhibitory effects within the cell wall structure constitute a serious drawback in maximizing the utilization of fibrous feedstuffs in animal feeding. Therefore treatments that promote efficient delignification of these materials must be applied. This study evaluated the potential of white-rot fungi to upgrade the nutritive value of cowpea stover for rabbit feeding. There was an increase in the crude protein content of all substrates as a result of fungi treatments, reaching a net gain of 13% for Pleurotus citrinopileatus incubation. Overall, net losses of dry and organic matter occurred during fungi treatments. Although the fiber content remained identical, higher consumption of cell wall contents was measured for P. citrinopileatus incubation (between 40 and 45%). The incubation period did not influence lignin degradation for any of the fungi treatments. Differences within the fungal degradation mechanisms indicate that P. citrinopileatus treatment was most effective, enhancing in vitro organic matter digestibility by around 30% compared with the control. Treatment of cowpea stover with P. citrinopileatus led to an efficient delignification process which resulted in higher in vitro organic matter digestibility, showing its potential in the nutritional valorization of this feedstuff. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Addition of allochthonous fungi to a historically contaminated soil affects both remediation efficiency and bacterial diversity.

PubMed

Federici, Ermanno; Leonardi, Vanessa; Giubilei, Maria A; Quaratino, Daniele; Spaccapelo, Roberta; D'Annibale, Alessandro; Petruccioli, Maurizio

2007-11-01

Botryosphaeria rhodina DABAC P82 and Pleurotus pulmonarius CBS 664.97 were tested for their ability to grow and to degrade aromatic hydrocarbons in an aged contaminated soil. To evaluate the impact of indigenous microflora on the overall process, incubations were performed on both fumigated and nonfumigated soils. Fungal colonization by B. rhodina was unexpectedly lower in the fumigated than in the nonfumigated soil while the growth of P. pulmonarius showed an opposite response. Degradation performances and detoxification by both fungi in the nonfumigated soil were markedly higher than those observed in the fumigated one. Heterotrophic bacterial counts in nonfumigated soil augmented with either B. rhodina or P. pulmonarius were significantly higher than those of the corresponding incubation control (6.7 +/- 0.3 x 10(8) and 8.35 +/- 0.6 x 10(8), respectively, vs 9.2 +/- 0.3 x 10(7)). Bacterial communities of both incubation controls and fungal-augmented soil were compared by numerical analysis of denaturing gradient gel electrophoresis profiles of polymerase chain reaction (PCR)-amplified 16S ribosomal RNA (rRNA) genes and cloning and sequencing of PCR-amplified 16S rRNA genes. Besides increasing overall diversity, fungal augmentation led to considerable qualitative differences with respect to the pristine soil.

Technical note: enumeration of mesophilic aerobes in milk: evaluation of standard official protocols and Petrifilm aerobic count plates.

PubMed

Freitas, R; Nero, L A; Carvalho, A F

2009-07-01

Enumeration of mesophilic aerobes (MA) is the main quality and hygiene parameter for raw and pasteurized milk. High levels of these microorganisms indicate poor conditions in production, storage, and processing of milk, and also the presence of pathogens. Fifteen raw and 15 pasteurized milk samples were submitted for MA enumeration by a conventional plating method (using plate count agar) and Petrifilm Aerobic Count plates (3M, St. Paul, MN), followed by incubation according to 3 official protocols: IDF/ISO (incubation at 30 degrees C for 72 h), American Public Health Association (32 degrees C for 48 h), and Brazilian Ministry of Agriculture (36 degrees C for 48 h). The results were compared by linear regression and ANOVA. Considering the results from conventional methodology, good correlation indices and absence of significant differences between mean counts were observed, independent of type of milk sample (raw or pasteurized) and incubation conditions (IDF/ISO, American Public Health Association, or Ministry of Agriculture). Considering the results from Petrifilm Aerobic Count plates, good correlation indices and absence of significant differences were only observed for raw milk samples. The microbiota of pasteurized milk interfered negatively with the performance of Petrifilm Aerobic Count plates, probably because of the presence of microorganisms that poorly reduce the dye indicator of this system.

Stochastic Modeling Approach to the Incubation Time of Prionic Diseases

NASA Astrophysics Data System (ADS)

Ferreira, A. S.; da Silva, M. A.; Cressoni, J. C.

2003-05-01

Transmissible spongiform encephalopathies are neurodegenerative diseases for which prions are the attributed pathogenic agents. A widely accepted theory assumes that prion replication is due to a direct interaction between the pathologic (PrPSc) form and the host-encoded (PrPC) conformation, in a kind of autocatalytic process. Here we show that the overall features of the incubation time of prion diseases are readily obtained if the prion reaction is described by a simple mean-field model. An analytical expression for the incubation time distribution then follows by associating the rate constant to a stochastic variable log normally distributed. The incubation time distribution is then also shown to be log normal and fits the observed BSE (bovine spongiform encephalopathy) data very well. Computer simulation results also yield the correct BSE incubation time distribution at low PrPC densities.

Effect of diet composition and incubation time on feed indigestible neutral detergent fiber concentration in dairy cows.

PubMed

Krizsan, S J; Huhtanen, P

2013-03-01

Indigestible neutral detergent fiber (NDF) predicts forage digestibility accurately and precisely when determined by a 288-h ruminal in situ incubation, and it is an important parameter in mechanistic rumen models. The long incubation time required is a disadvantage. Further, intrinsic cell wall characteristics of feeds should be determined under ideal conditions for fiber digestion. The objective of this study was to determine the effects of diet composition and rumen incubation time on the concentrations of indigestible NDF (iNDF) for a wide range of feeds in dairy cows. Additionally, predicted concentrations of unavailable NDF generated using the National Research Council (NRC) model and the Cornell Net Carbohydrate and Protein System (CNCPS) were evaluated. Indigestible NDF was evaluated in 18 feeds using 4 cows in a split-split plot design. Treatments were in a 3 × 3 factorial arrangement, consisting of different diets and incubation times. Diet composition was primarily varied by changing the level of concentrate supplementation between 190 (low), 421 (medium), and 625 (high)g/kg of diet dry matter (DM). Grass silage was used as the basal forage for all cows. The feeds were incubated for 144, 216, and 288 h. Indigestible NDF was determined from 2-g samples weighed into polyester bags with a pore size of 12 µm and a pore area equal to 6% of the total surface area, giving a sample size to surface ratio of 10mg/cm(2). Across all feeds, the measured iNDF concentrations ranged from 6 to 516 g/kg of DM. The feed iNDF concentration was not affected by the cow used, but diet composition had a significant effect. The mean measured iNDF concentrations for cows consuming low-, medium-, and high-concentrate diets were 178, 186, and 197 g/kg of DM, respectively. The incubation time also affected the feed iNDF concentrations, which averaged 199, 185, and 177 g/kg of DM for 144-, 216-, and 288-h incubations, respectively. We also observed significant interactions between incubation time and feed, and between diet composition and feed, with fiber-rich feeds being most sensitive to these factors. The evaluation of model predictions of unavailable NDF indicated poor precision with prediction errors of 56 (NRC) and 84 (CNCPS)g/kg of DM. Indigestible NDF should be determined based on 288-h ruminal in situ incubations in cows consuming diets with a low proportion of concentrate to represent the feed fraction that is unavailable to the animal. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

A salivary incubation test for evaluation of oral malodor: a pilot study.

PubMed

Quirynen, Marc; Zhao, Hong; Avontroodt, Pieter; Soers, Catherine; Pauwels, Martine; Coucke, Wim; van Steenberghe, Daniel

2003-07-01

Breath odor is scored by different techniques, each with its own shortcomings. Organoleptic ratings are uncomfortable for the patient, subjective, influenced by external parameters including food and cosmetics, and especially lack international calibration. Portable sulphide monitors are relatively expensive and neglect several major malodorous molecules (e.g., butyric and propionic acids, putrescine, and cadaverine). Gas chromatography necessitates expensive devices and experienced technicians. This pilot study explored the applicability of a new technique (saliva incubation) by comparing its discrimination power, in a morning bad breath inhibition study of antiseptics, to those of hydrogen sulphide (H2S) measurement devices and organoleptic ratings. After a professional cleaning, 8 periodontally healthy students abstained from all means of mechanical plaque control for 5 experimental periods of 7 days, with intervening washout periods of at least 2 weeks. During each experimental period, the students rinsed only twice daily with different antiseptics. At day 7, morning breath was scored clinically (volatile sulphide compound [VSC] level and organoleptic ratings), and 1.5 ml of saliva was collected and divided between 3 glass tubes that were sealed and incubated (37 degrees C, anaerobic chamber). Immediately after collection and after 3 and 6 hours of incubation, the headspace air in one of the tubes was examined for VSC production and organoleptic measurements. The investigations of the incubated saliva correlated well with the 7-day intraoral VSC recordings and organoleptic ratings (P < or = 0.005). Moreover, evaluations showed a similar interproduct ranking for their efficacy in malodor control. The power analyses indicated a higher discrimination power for the saliva incubation test than for the intraoral registrations. The strong correlation between odor production of incubated saliva and clinical assessments suggests that the saliva incubation test may be used as an indirect method to measure oral malodor and can be employed to investigate the antimalodor effectiveness of oral hygiene products.

9 CFR 147.23 - Hatchery sanitation.

Code of Federal Regulations, 2013 CFR

2013-01-01

... rooms are provided for each of the four operations: Egg receiving, incubation and hatching, chick/poult processing, and egg tray and hatching basket washing. Traffic and airflow patterns in the hatchery should be... should be cleaned and sanitized after each use. (d) The hatching compartments of incubators, including...

9 CFR 147.23 - Hatchery sanitation.

Code of Federal Regulations, 2012 CFR

2012-01-01

... rooms are provided for each of the four operations: Egg receiving, incubation and hatching, chick/poult processing, and egg tray and hatching basket washing. Traffic and airflow patterns in the hatchery should be... should be cleaned and sanitized after each use. (d) The hatching compartments of incubators, including...

9 CFR 147.23 - Hatchery sanitation.

Code of Federal Regulations, 2014 CFR

2014-01-01

... rooms are provided for each of the four operations: Egg receiving, incubation and hatching, chick/poult processing, and egg tray and hatching basket washing. Traffic and airflow patterns in the hatchery should be... should be cleaned and sanitized after each use. (d) The hatching compartments of incubators, including...

Cost assessment of the automated VERSANT 440 Molecular System versus the semi-automated System 340 bDNA Analyzer platforms.

PubMed

Elbeik, Tarek; Loftus, Richard A; Beringer, Scott

2007-11-01

Labor, supply and waste were evaluated for HIV-1 and HCV bDNA on the semi-automated System 340 bDNA Analyzer and the automated VERSANT 440 Molecular System (V440). HIV-1 sample processing was evaluated using a 24- and 48-position centrifuge rotor. Vigilance time (hands-on manipulations plus incubation time except initial target hybridization) and disposables were approximately 37 and 12% lower for HIV-1, and 64 and 31% lower for HCV bDNA, respectively, with V440. Biohazardous solid waste was approximately twofold lower for both assays and other waste types were the same for either assay on both platforms. HIV-1 sample processing vigilance time for the 48-position rotor was reduced by 2 h. V440 provides cost savings and improved workflow.

Chemical controls on abiotic and biotic release of geogenic arsenic from Pleistocene aquifer sediments to groundwater.

PubMed

Gillispie, Elizabeth C; Andujar, Erika; Polizzotto, Matthew L

2016-08-10

Over 150 million people in South and Southeast Asia consume unsafe drinking water from arsenic-rich Holocene aquifers. Although use of As-free water from Pleistocene aquifers is a potential mitigation strategy, such aquifers are vulnerable to geogenic As pollution, placing millions more people at potential risk. The goal of this research was to define chemical controls on abiotic and biotic release of geogenic As to groundwater. Batch incubations of sediments with natural chemical variability from a Pleistocene aquifer in Cambodia were conducted to evaluate how interactions among arsenic, manganese and iron oxides, and dissolved and sedimentary organic carbon influenced As mobilization from sediments. The addition of labile dissolved organic carbon produced the highest concentrations of dissolved As after >7 months, as compared to sediment samples incubated with sodium azide or without added carbon, and the extent of As release was positively correlated with the percent of initial extractable Mn released from the sediments. The mode of As release was impacted by the source of DOC supplied to the sediments, with biological processes responsible for 81% to 85% of the total As release following incubations with lactate and acetate but only up to 43% to 61% of the total As release following incubations with humic and fulvic acids. Overall, cycling of key redox-active elements and organic-carbon reactivity govern the potential for geogenic As release to groundwater, and results here may be used to formulate better predictions of the arsenic pollution potential of aquifers in South and Southeast Asia.

Incubation temperature and hemoglobin dielectric of chicken embryos incubated under the influence of electric field.

PubMed

Shafey, T M; Al-Batshan, H A; Shalaby, M I; Ghannam, M M

2006-01-01

Eggs from a layer-type breeder flock (Baladi, King Saud University) between 61 and 63 weeks of age were used in 3 trials to study the effects of electric field (EF) during incubation on the internal temperature of incubation, and eggs and hemoglobin (Hb) dielectric of chicken embryos at 18 days of age. Dielectric relative permittivity (epsilon') and conductivity (sigma) of Hb were examined in the range of frequency from 20 to 100 kHz. The values of dielectric increment (Deltaepsilon') and the relaxation times (tau) of Hb molecules were calculated. The internal temperature of eggs was measured in empty (following the removal of egg contents) and fertilized eggs in trials 1 and 2, respectively. The level of the EF was 30 kV/m, 60 Hz. EF incubation of embryos influenced the temperature of incubation and electrical properties of Hb molecules and did not influence the temperature of incubation and internal environment of eggs when empty eggs were incubated. EF incubation of fertilized eggs significantly raised the temperature of incubation, egg air cell, and at the surface of the egg yolk by approximately 0.09, 0.60, and 0.61 degrees F, respectively and Hb epsilon', sigma, Deltaepsilon', and tau as a function of the range of frequency of 20 to 100 kHz when compared with their counterparts of the control group. It was concluded that the exposure of fertilized chicken eggs to EF of 30 kV/m, 60 Hz, during incubation altered dielectric properties of Hb and that probably affected cell to cell communication and created the right environment for enhancing the growing process and heat production of embryos consequently increasing the temperature of the internal environment of the egg, and incubation.

Humidity control of an incubator using the microcontroller-based active humidifier system employing an ultrasonic nebulizer.

PubMed

Güler, I; Burunkaya, M

2002-01-01

Relative humidity levels of an incubator were measured and controlled. An ultrasonic nebulizer system as an active humidifier was used to humidify the incubator environment. An integrated circuit-type humidity sensor was used to measure the humidity level of the incubator environment. Measurement and control processes were achieved by a PIC microcontroller. The high-performance and high-speed PIC provided the flexibility of the system. The developed system can be used effectively for the intensive care of newborns and/or premature babies. Since the humidifier generates an aerosol in ambient conditions, it is possible to provide the high relative humidity level for therapeutic and diagnostic purposes in medicine.

How pH, Temperature, and Time of Incubation Affect False-Positive Responses and Uncertainty of the LAL Gel-Clot Test.

PubMed

Lourenço, Felipe Rebello; Botelho, Túlia De Souza; Pinto, Terezinha De Jesus Andreoli

2012-01-01

The limulus amebocyte lysate (LAL) test is the simplest and most widely used procedure for detection of endotoxin in parenteral drugs. The LAL test demands optimal pH, ionic strength, temperature, and time of incubation. Slight changes in these parameters may increase the frequency of false-positive responses and the estimated uncertainty of the LAL test. The aim of this paper is to evaluate how changes in the pH, temperature, and time of incubation affect the occurrence of false-positive responses in the LAL test. LAL tests were performed in nominal conditions (37 °C, 60 min, and pH 7) and in different conditions of temperature (36 °C and 38 °C), time of incubation (58 and 62 min), and pH (6 and 8). Slight differences in pH increase the frequency of false-positive responses 5-fold (relative risk 5.0), resulting in an estimated of uncertainty 7.6%. Temperature and time of incubation affect the LAL test less, showing relative risks of 1.5 and 1.0, respectively. Estimated uncertainties in 36 °C or 38 °C temperatures and 58 or 62 min of incubation were found to be 2.0% and 1.0%, respectively. Simultaneous differences in these parameters significantly increase the frequency of false-positive responses. The limulus amebocyte lysate (LAL) gel-clot test is a simple test for detection of endotoxin from Gram-negative bacteria. The test is based on a gel formation when a certain amount of endotoxin is present; it is a pass/fail test. The LAL test requires optimal pH, ionic strength, temperature, and time of incubation. Slight difference in these parameters may increase the frequency of false-positive responses. The aim of this paper is to evaluate how changes in the pH, temperature, and time of incubation affect the occurrence of false-positive responses in the LAL test. We find that slight differences in pH increase the frequency of false-positive responses 5-fold. Temperature and time of incubation affect the LAL test less. Simultaneous differences in these parameters significantly increase the frequency of false-positive responses.

Indirect antiglobulin test-crossmatch using low-ionic-strength saline-albumin enhancement medium and reduced incubation time: effectiveness in the detection of most clinically significant antibodies and impact on blood utilization.

PubMed

Dinardo, C L; Bonifácio, S L; Mendrone, A

2014-01-01

Indirect antiglobulin test-crossmatch (IAT-XM) using enhancement media such as low-ionic-strength saline (LISS) and polyethylene glycol (PEG) usually requires 15 minutes of incubation. These methods are necessary when testing samples from blood recipients who have a higher risk of alloimmunization. In emergency situations, IAT-XM can be time-consuming and can influence presurgery routine, resulting in more red blood cell (RBC) units being tested and stored to avoid the transfusion of uncrossmatched ones. The objective of this study was to evaluate the performance of a LISS-albumin enhancer to intensify antigen-antibody reaction after 5 minutes of 37oC incubation and compare this performance with that of other enhancers, gel, and conventional tube testing. Second, the study evaluated the impact of this method's implementation in the C:T ratio (crossmatched to transfused RBC units) of a transfusion laboratory. Ninety serum samples containing alloantibodies of potential clinical significance were tested against phenotyped RBCs using four different methods: (1) tube with LISS-albumin enhancer (5 minutes of incubation), (2) tube with LISS-albumin and PEG (15 minutes of incubation), (3) gel, and (4) conventional tube method (60 minutes of incubation). In parallel, the study compared the C:T ratio of a tertiary-hospital transfusion laboratory in two different periods: 3 months before and 3 months after the implementation of the 5-minute IAT-XM protocol. The use of LISS-albumin with 5 minutes of incubation exhibited the same performance as LISS-albumin, PEG, and gel with 15 minutes of incubation. Conventional tube method results were equally comparable, but reactions were significantly less intense, except for anti-c (p = 0.406). Accuracy was 100 percent for all selected methods. After the implementation of the 5-minute IAT-XM protocol, the C:T ratio fell from 2.74 to 1.29 (p < 0.001). IAT-XM can have its incubation time reduced to 5 minutes with the use of LISS-albumin enhancement. We suggest this strategy should be used to quickly prepare RBC units for surgical patients, keeping transfusion safety without compromising blood supplies.

Vitamins E and C Prevent DNA Double-strand Breaks in Peripheral Lymphocytes Exposed to Radiations from Iodine-131.

PubMed

Safaei, Mehdi; Jafarpour, Seyed Masoud; Mohseni, Mehran; Salimian, Morteza; Akbari, Hossein; Karami, Fateme; Aliasgharzadeh, Akbar; Farhood, Bagher

2018-01-01

Iodine-131 is used as a radiopharmaceutical to treat thyroid cancer. The current study aimed to evaluate the effects of vitamins E and C on the level of DNA double-strand breaks (DSBs) caused by Radioiodine-131 (I-131) in human lymphocytes. Whole blood samples from human volunteers were incubated with a certain concentration of vitamins. After 1-h incubation, the samples were incubated with 20 μCi I-131/2 mL (blood + NaCl) for 1 h. To evaluate the effects of antioxidants, lymphocytes were separated, and the mean DSBs/cell was measured for each sample through γ-H2AX assay. After 1-h incubation with 20 μCi I-131/2 mL (blood + NaCl), iodine-131 increased the level of DSBs by 102.9%, compared with the background group. Vitamins E and C reduced the level of DSBs by 21.5% and 36.4%, respectively. Using vitamins E and C as antioxidants can reduce the toxicity of I-131. Furthermore, vitamin C provided the more protection for DNA, compared with vitamin E.

Evaluation of noise in the neonatal intensive care unit.

PubMed

Benini, F; Magnavita, V; Lago, P; Arslan, E; Pisan, P

1996-01-01

This study evaluated the noise level inside the incubators in a neonatal intensive care unit and identified its sources in order to attempt to reduce it. Although noise is not a proven risk factor as far as the sensory integrity of newborns is concerned, it is certainly an important cause of stress to them and a source of serious and dangerous changes in their behavioral and physiologic states. Noise recorded inside the incubators had two components. The first was background noise from the incubator motors, which varied from 74.2 to 79.9 dB, and was similar to environmental noise. The second source was impulsive events beyond 80 dB. These events were the result of voluntary and involuntary contact with the incubators' Plexiglas surface or to the abrupt opening and closing of their access ports. Considering its decibel levels and frequency, this latter component is undoubtedly an important source of stress to newborns. Moreover, these data reveal the need to train health care personnel on how to reduce such noise by taking more care in the handling of infants.

Plasma biochemistry values in emperor geese (Chen canagica) in Alaska: comparisons among age, sex, incubation, and molt.

USGS Publications Warehouse

Franson, J. Christian; Hoffman, D.J.; Schmutz, J.A.

2009-01-01

Reduced populations of emperor geese (Chen canagica), a Bering Sea endemic, provided the need to assess plasma biochemistry values as indicators of population health. A precursory step to such an investigation was to evaluate patterns of variability in plasma biochemistry values among age, sex, and reproductive period. Plasma from 63 emperor geese was collected on their breeding grounds on the Yukon-Kuskokwim Delta in western Alaska, USA. The geese sampled included 18 incubating adult females captured, in mid June, on their nests by using bow nets, and 30 adults and 15 goslings captured in corral traps in late July and early August, when the adults were molting their wing feathers and the goslings were 5-6 weeks old. Plasma was evaluated for 15 biochemical parameters, by comparing results among age, sex, and sampling period (incubation versus wing-feather molt). Ten of the 15 biochemical parameters assayed differed among adults during incubation, the adults during molt, and the goslings at molt, whereas sex differences were noted in few parameters.

Determining the need for improvement of infant incubator design with quality function deployment

NASA Astrophysics Data System (ADS)

Tarigan, U.; Ginting, R.; Siregar, I.

2018-02-01

A Newborns need particular attention because it takes time to adapt to the outside world. In accordance with the standard post-neonatal procedures, newborns should be incorporated into the incubator within a specified period of time according to the infant’s health level. Infant incubator as a medical device used to care for newborns, often get complaints from doctors and child nurses. The complaint indicates consumer dissatisfaction with incubator products in the hospital. Broadly speaking, objection against infant incubators lie in inappropriate designs. To overcome these complaints the researchers apply the method of Quality Function Deployment to determine the characteristics of priority techniques in accordance with the wishes of consumers with it. The primary focus of QFD is to engage customers in the product development process as early as possible, which their needs and desires serve as the starting point of the QFD process. Therefore, QFD is called the voice of customer. The underlying philosophy is that customers are not always satisfied with a product even though the product has been perfectly produced. The results show that the category used as a priority improvement is the additional function of the oxygen cylinder and the size of the door hole. QFD phase one produces technical characteristics of Ergonomy, Features and Cost as critical part determinants.

Human renal adipose tissue induces the invasion and progression of renal cell carcinoma

PubMed Central

Campo-Verde-Arbocco, Fiorella; López-Laur, José D.; Romeo, Leonardo R.; Giorlando, Noelia; Bruna, Flavia A.; Contador, David E.; López-Fontana, Gastón; Santiano, Flavia E.; Sasso, Corina V.; Zyla, Leila E.; López-Fontana, Constanza M.; Calvo, Juan C.; Carón, Rubén W.; Creydt, Virginia Pistone

2017-01-01

We evaluated the effects of conditioned media (CMs) of human adipose tissue from renal cell carcinoma located near the tumor (hRATnT) or farther away from the tumor (hRATfT), on proliferation, adhesion and migration of tumor (786-O and ACHN) and non-tumor (HK-2) human renal epithelial cell lines. Human adipose tissues were obtained from patients with renal cell carcinoma (RCC) and CMs from hRATnT and hRATfT incubation. Proliferation, adhesion and migration were quantified in 786-O, ACHN and HK-2 cell lines incubated with hRATnT-, hRATfT- or control-CMs. We evaluated versican, adiponectin and leptin expression in CMs from hRATnT and hRATfT. We evaluated AdipoR1/2, ObR, pERK, pAkt y pPI3K expression on cell lines incubated with CMs. No differences in proliferation of cell lines was found after 24 h of treatment with CMs. All cell lines showed a significant decrease in cell adhesion and increase in cell migration after incubation with hRATnT-CMs vs. hRATfT- or control-CMs. hRATnT-CMs showed increased levels of versican and leptin, compared to hRATfT-CMs. AdipoR2 in 786-O and ACHN cells decreased significantly after incubation with hRATfT- and hRATnT-CMs vs. control-CMs. We observed a decrease in the expression of pAkt in HK-2, 786-O and ACHN incubated with hRATnT-CMs. This result could partially explain the observed changes in migration and cell adhesion. We conclude that hRATnT released factors, such as leptin and versican, could enhance the invasive potential of renal epithelial cell lines and could modulate the progression of the disease. PMID:29212223

Incubation Temperature, But Not Pequi Oil Supplementation, Affects Methane Production, and the Ruminal Microbiota in a Rumen Simulation Technique (Rusitec) System.

PubMed

Duarte, Andrea C; Holman, Devin B; Alexander, Trevor W; Kiri, Kerstin; Breves, Gerhard; Chaves, Alexandre V

2017-01-01

Lipid supplementation is a promising strategy for methane mitigation in cattle and has been evaluated using several different lipid sources. However, limited studies have assessed the effect of temperature on methane emissions from cattle and changes in incubation temperature have also not been extensively evaluated. The aim of this study was to evaluate the combined effect of pequi oil (high in unsaturated fatty acids) and incubation temperature on fermentation characteristics and microbial communities using the rumen simulation technique. A completely randomized experiment was conducted over a 28-day period using a Rusitec system. The experiment was divided into four periods of 7 days each, the first of which was a 7-day adaptation period followed by three experimental periods. The two treatments consisted of a control diet (no pequi oil inclusion) and a diet supplemented with pequi oil (1.5 mL/day) which increased the dietary fat content to 6% (dry matter, DM-basis). Three fermenter vessels (i.e., replicates) were allocated to each treatment. In the first experimental period, the incubation temperature was maintained at 39°C, decreased to 35°C in the second experimental period and then increased again to 39°C in the third. Pequi oil was continuously supplemented during the experiment. Microbial communities were assessed using high-throughput sequencing of the archaeal and bacterial 16S rRNA gene. Methane production was reduced by 57% following a 4°C decrease in incubation temperature. Supplementation with pequi oil increased the dietary fat content to 6% (DM-basis) but did not affect methane production. Analysis of the microbiota revealed that decreasing incubation temperature to 35°C affected the archaeal and bacterial diversity and richness of liquid-associated microbes, but lipid supplementation did not change microbial diversity.

Toxicological evaluation of some Malaysian locally processed raw food products.

PubMed

Sharif, R; Ghazali, A R; Rajab, N F; Haron, H; Osman, F

2008-01-01

Malaysian locally processed raw food products are widely used as main ingredients in local cooking. Previous studies showed that these food products have a positive correlation with the incidence of cancer. The cytotoxicity effect was evaluated using MTT assay (3-(4,5-dimetil-2-thiazolil)-2,5-diphenyl-2H-tetrazolium bromide) against Chang liver cells at 2000 microg/ml following 72 h incubation. Findings showed all methanol extracts caused a tremendous drop in the percentage of cell viability at 2000 microg/ml (shrimp paste - 41.69+/-3.36%, salted fish - 37.2+/-1.06%, dried shrimp - 40.32+/-1.8%, p<0.05). To detect DNA damage in a single cell, alkaline Comet Assay was used. None of the extracts caused DNA damage to the Chang liver cells at 62.5 microg/ml following 24 h incubation, as compared to the positive control, hydrogen peroxide (tail moment - 9.50+/-1.50; tail intensity - 30.50+/-2.50). Proximate analysis which was used for the evaluation of macronutrients in food showed that shrimp paste did not comply with the protein requirement (<25%) as in Food Act 1983. Salt was found in every sample with the highest percentage being detected in shrimp paste which exceeded 20%. Following heavy metal analysis (arsenic, cadmium, lead and mercury), arsenic was found in every sample with dried shrimps showing the highest value as compared to the other samples (6.16 mg/kg). In conclusion, several food extracts showed cytotoxic effect but did not cause DNA damage against Chang liver cells. Salt was found as the main additive and arsenic was present in every sample, which could be the probable cause of the toxicity effects observed.

Effect of tilmicosin on chemotactic, phagocytic, and bactericidal activities of bovine and porcine alveolar macrophages.

PubMed

Brumbaugh, Gordon W; Herman, James D; Clancy, Julianne S; Burden, Kyland I; Barry, Tracie; Simpson, R B; López, Hector Sumano

2002-01-01

To evaluate chemotactic, phagocytic, and bactericidal activities of bovine and porcine alveolar macrophages (AM) exposed to tilmicosin. 12 healthy calves and 12 healthy pigs. Lungs were obtained immediately after euthanasia; AM were collected by means of bronchoalveolar lavage and density gradient centrifugation. Chemotactic activity was evaluated by exposing AM to lipopolysaccharide or macrophage inhibitory peptide during incubation with tilmicosin. Phagocytic activity was evaluated by incubating AM with tilmicosin for 24 hours and then with tilmicosin-resistant Salmonella serotype Typhimurium. Bactericidal activity was evaluated by incubating AM with tilmicosin (0, 10, or 20 microg/ml for bovine AM; 0 or 10 microg/ml or 10 microg/ml but washed free of tilmicosin for porcine AM) and then with Mannheimia haemolytica (bovine AM) or with Actinobacillus pleuropneumoniae or Pasteurella multocida (porcine AM). Tilmicosin had no significant effects on chemotactic or phagocytic activities of bovine or porcine AM. The time-course of bactericidal activity was best described by polynomial equations. Time to cessation of bacterial growth and area under the time versus bacterial number curve were significantly affected by incubation of AM with tilmicosin. Results show that bactericidal activity of bovine and porcine AM was enhanced by tilmicosin, but not in proportion to the reported ability of AM to concentrate tilmicosin intracellularly. With or without exposure to tilmicosin, the time-course of bactericidal activity of bovine AM against M haemolytica and of porcine AM against A pleuropneumoniae or P multocida was too complex to be reduced to a simple linear equation.

Stabilization by hydrophobic protection as a molecular mechanism for organic carbon sequestration in maize-amended rice paddy soils.

PubMed

Song, X Y; Spaccini, R; Pan, G; Piccolo, A

2013-08-01

The hydrophobic components of soil organic matter (SOM) are reckoned to play an important role in the stabilization of soil organic carbon (SOC). The contribution of hydrophobic substances to SOC sequestration was evaluated in four different paddy soils in the South of China, following a 6-month incubation experiment with maize straw amendments. Soil samples included: a well developed paddy soil (TP) derived from clayey lacustrine deposits in the Tai Lake plain of Jiangsu; an acid clayey paddy soil (RP) derived from red earth in the rolling red soil area of Jiangxi; a weakly developed neutral paddy soil (PP) formed on Jurassic purple shale from Chongq; and a calcic Fluvisol (MS) derived from riverine sediments from a wetland along the Yangtze valley of Anhui, China. The SOC molecular composition after 30 and 180 days of incubation, was determined by off-line thermochemolysis followed by gas chromatography-mass spectrometry analysis. Lignin, lipids and carbohydrates were the predominant thermochemolysis products released from the treated soils. A selective preservation of hydrophobic OM, including lignin and lipids, was shown in maize amended soils with prolonged incubation. The decomposition of lignin and lipids was significantly slower in the TP and RP soils characterized by a larger content of extractable iron oxyhydrates (Fed) and lower pH. The overall increase in hydrophobic substances in maize incubated samples was correlated, positively, with total content of clay and Fed, and, negatively, with soil pH. Moreover, yields of both lignin and lipid components showed a significant relationship with SOC increase after incubation. These findings showed that the larger the lipid and lignin content of SOM, the greater was the stability of SOC, thereby suggesting that OM hydrophobic components may have an essential role in controlling the processes of OC sequestration in paddy soils of South China. Copyright © 2013 Elsevier B.V. All rights reserved.

Incubation Patterns in a Central-Place Forager Affect Lifetime Reproductive Success: Scaling of Patterns from a Foraging Bout to a Lifetime

PubMed Central

Shoji, Akiko; Elliott, Kyle H.; Aris-Brosou, Stéphane; Crump, Doug; Gaston, Anthony J.

2011-01-01

Background Long-lived seabirds face a conflict between current and lifelong reproductive success. During incubation shifts, egg neglect is sometimes necessary to avoid starvation, but may compromise the current reproductive attempt. However, factors underlying this decision process are poorly understood. We focus on the ancient murrelet, Synthliboramphus antiquus, an alcid with exceptionally long incubation shift lengths, and test the impact of environmental factors on incubation shift length in relation to reproductive success. Methodology/Principal Findings Using an information theoretic approach, we show that incubation shift length was a strong predictor of reproductive success for ancient murrelets at Reef Island, Haida Gwaii, British Columbia, Canada during the 2007 and 2008 breeding seasons. The most important factors explaining an individual's shift length were egg size, wind speed and the length of the mate's previous shift. Wind speed and tide height were the two most important factors for determining foraging behavior, as measured by dive frequency and depth. Conclusions/Significance Our study demonstrates that (i) species-specific reproductive strategies interact with environmental conditions such as wind speed to form multiple incubation patterns and (ii) maintaining regular incubation shifts is an essential component of reproductive success. PMID:21423631

High Environmental Temperature Increases Glucose Requirement in the Developing Chicken Embryo

PubMed Central

Molenaar, Roos; van den Borne, Joost J. G. C.; Hazejager, Ewoud; Kristensen, Niels B.; Heetkamp, Marcel J. W.; Meijerhof, Ron; Kemp, Bas; van den Brand, Henry

2013-01-01

Environmental conditions during the perinatal period influence metabolic and developmental processes in mammals and avian species, which could impact pre- and postnatal survival and development. The current study investigated the effect of eggshell temperature (EST) on glucose metabolism in broiler chicken embryos. Broiler eggs were incubated at a high (38.9°C) or normal (37.8°C) EST from day 10.5 of incubation onward and were injected with a bolus of [U-13C]glucose in the chorio-allantoic fluid at day 17.5 of incubation. After [U-13C]glucose administration, 13C enrichment was determined in intermediate pools and end-products of glucose metabolism. Oxidation of labeled glucose occurred for approximately 3 days after injection. Glucose oxidation was higher in the high than in the normal EST treatment from day 17.6 until 17.8 of incubation. The overall recovery of 13CO2 tended to be 4.7% higher in the high than in the normal EST treatment. An increase in EST (38.9°C vs 37.8°C) increased 13C enrichment in plasma lactate at day 17.8 of incubation and 13C in hepatic glycogen at day 18.8 of incubation. Furthermore, high compared to normal EST resulted in a lower yolk-free body mass at day 20.9 (−2.74 g) and 21.7 (−3.81 g) of incubation, a lower hepatic glycogen concentration at day 18.2 (−4.37 mg/g) and 18.8 (−4.59 mg/g) of incubation, and a higher plasma uric acid concentration (+2.8 mg/mL/+43%) at day 21.6 of incubation. These results indicate that the glucose oxidation pattern is relatively slow, but the intensity increased consistently with an increase in developmental stage of the embryo. High environmental temperatures in the perinatal period of chicken embryos increased glucose oxidation and decreased hepatic glycogen prior to the hatching process. This may limit glucose availability for successful hatching and could impact body development, probably by increased gluconeogenesis from glucogenic amino acids to allow anaerobic glycolysis. PMID:23560054

High environmental temperature increases glucose requirement in the developing chicken embryo.

PubMed

Molenaar, Roos; van den Borne, Joost J G C; Hazejager, Ewoud; Kristensen, Niels B; Heetkamp, Marcel J W; Meijerhof, Ron; Kemp, Bas; van den Brand, Henry

2013-01-01

Environmental conditions during the perinatal period influence metabolic and developmental processes in mammals and avian species, which could impact pre- and postnatal survival and development. The current study investigated the effect of eggshell temperature (EST) on glucose metabolism in broiler chicken embryos. Broiler eggs were incubated at a high (38.9°C) or normal (37.8°C) EST from day 10.5 of incubation onward and were injected with a bolus of [U-(13)C]glucose in the chorio-allantoic fluid at day 17.5 of incubation. After [U-(13)C]glucose administration, (13)C enrichment was determined in intermediate pools and end-products of glucose metabolism. Oxidation of labeled glucose occurred for approximately 3 days after injection. Glucose oxidation was higher in the high than in the normal EST treatment from day 17.6 until 17.8 of incubation. The overall recovery of (13)CO2 tended to be 4.7% higher in the high than in the normal EST treatment. An increase in EST (38.9°C vs 37.8°C) increased (13)C enrichment in plasma lactate at day 17.8 of incubation and (13)C in hepatic glycogen at day 18.8 of incubation. Furthermore, high compared to normal EST resulted in a lower yolk-free body mass at day 20.9 (-2.74 g) and 21.7 (-3.81 g) of incubation, a lower hepatic glycogen concentration at day 18.2 (-4.37 mg/g) and 18.8 (-4.59 mg/g) of incubation, and a higher plasma uric acid concentration (+2.8 mg/mL/+43%) at day 21.6 of incubation. These results indicate that the glucose oxidation pattern is relatively slow, but the intensity increased consistently with an increase in developmental stage of the embryo. High environmental temperatures in the perinatal period of chicken embryos increased glucose oxidation and decreased hepatic glycogen prior to the hatching process. This may limit glucose availability for successful hatching and could impact body development, probably by increased gluconeogenesis from glucogenic amino acids to allow anaerobic glycolysis.

Aerobic biodegradation of sludge with high hydrocarbon content generated by a Mexican natural gas processing facility.

PubMed

Roldán-Carrillo, T; Castorena-Cortés, G; Zapata-Peñasco, I; Reyes-Avila, J; Olguín-Lora, P

2012-03-01

The biodegradation of oil sludge from Mexican sour gas and petrochemical facilities contaminated with a high content of hydrocarbons, 334.7 ± 7.0 g kg(-1) dry matter (dm), was evaluated. Studies in microcosm systems were carried out in order to determine the capacity of the native microbiota in the sludge to reduce hydrocarbon levels under aerobic conditions. Different carbon/nitrogen/phosphorous (C/N/P) nutrient ratios were tested. The systems were incubated at 30 °C and shaken at 100 rpm. Hydrocarbon removals from 32 to 51% were achieved in the assays after 30 days of incubation. The best assay had C/N/P ratio of 100/1.74/0.5. The results of the Microtox(®) and Ames tests indicated that the original sludge was highly toxic and mutagenic, whereas the best assay gave a final product that did not show toxicity or mutagenicity. Copyright © 2011 Elsevier Ltd. All rights reserved.

Interactions between organic amendments and phosphate fertilizers modify phosphate sorption processes in an acid soil

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sckefe, C.R.; Patti, A.F.; Clune, T.S.

2008-07-15

To determine how organic amendments and phosphate fertilizers interact to modify P sorption processes, three phosphate fertilizers were applied to lignite- and compost-amended acid soil and incubated for either 3 or 26 days. The fertilizers applied were potassium dihydrogen phosphate, triple superphosphate, and diammonium phosphate (DAP). After 3 days of incubation, sorption of all three P sources was decreased in the lignite-amended treatments, whereas P sorption was increased in the compost-amended treatments. Increased incubation time (26 days) resulted in significantly decreased P sorption when DAP was added to lignite-amended treatments. Addition of triple superphosphate increased P sorption in lignite- andmore » compost-amended treatments and decreased solution pH compared with DAP application. In addition to the effect of P source, differences in P sorption between the lignite- and compost-amended treatments were driven by differences in solution chemistry, predominantly solution pH and cation dynamics. Soil amendment and fertilizer addition also increased microbial activity in the incubation systems, as measured by carbon dioxide respiration. It is proposed that the combination of lignite and DAP may contribute to decreased P sorption in acid soils, with the positive effects likely caused by both chemical and biological processes, including the formation of soluble organic-metal complexes.« less

Incubation temperature effects on physical characteristics of normal, dark, firm and dry, and halothane-carrier pork longissimus.

PubMed

McCaw, J; Ellis, M; Brewer, M S; McKeith, F K

1997-06-01

Pigs (n = 18) were selected to represent three different muscle conditions (six pigs per condition): normal: dark, firm, and dry; and halothane carrier. A 45-cm-long longissimus section was excised from each side of the carcass at 30 min postmortem and cut into six sections. Right side sections were assigned to the intermediate temperature incubation (23 degrees C), and left side sections were designated high temperature incubation (40 degrees C). Sections were randomly assigned to incubation times (0, 1, 2, 4, 6, or 8 h). The 0 h section from each incubation treatment was designated as a control and was placed directly into a 4 degree C cooler. Temperature and pH were evaluated on the control section and for each loin section a the end of the incubation time. Color (L*, a*, and b* values), percentage of purge loss, water-holding capacity, and drip loss were determined. Incubation treatment did not alter pH decline in dark, firm, and dry muscle; however, high temperature increased pH decline in normal and halothane carrier samples. Results suggest that there is a strong interaction between pH and temperature that affects pork quality attributes. High incubation temperature had a negative effect on most quality variables; however, muscle condition (normal or halothane carrier) had limited effects on muscle quality.

Mind Wandering and the Incubation Effect in Insight Problem Solving

ERIC Educational Resources Information Center

Tan, Tengteng; Zou, Hong; Chen, Chuansheng; Luo, Jin

2015-01-01

Although many anecdotes suggest that creative insights often arise during mind wandering, empirical research is still sparse. In this study, the number reduction task (NRT) was used to assess whether insightful solutions were related to mind wandering during the incubation stage of the creative process. An experience sampling paradigm was used to…

Conversion of canola meal into a high-protein feed additive via solid-state fungal incubation process

USDA-ARS?s Scientific Manuscript database

The study goal was to determine the optimal fungal culture to reduce glucosinolates (GLS), fiber, and residual sugars while increasing the protein content and nutritional value of canola meal. Solid-state incubation conditions were used to enhance filamentous growth of the fungi. Flask trials were p...

Incubation reduces microbial growth on eggshells and the opportunity for trans-shell infection.

Treesearch

Mark I. Cook; Steven R. Beissinger; Gary A. Toranzos; Wayne J. Arendt

2005-01-01

Avian eggshells harbour microbes shortly after laying, and under appropriate ambient conditions they can multiply rapidly, penetrate through shell pores, infect egg contents and cause embryo mortality. We experimentally examined how incubation affects bacterial processes on the eggshells of pearl-eyed thrashers Margarops fuscatus nesting in tropical montane and lowland...

Study of aliphatic-aromatic copolyester degradation in sandy soil and its ecotoxicological impact.

PubMed

Rychter, Piotr; Kawalec, Michał; Sobota, Michał; Kurcok, Piotr; Kowalczuk, Marek

2010-04-12

Degradation of poly[(1,4-butylene terephthalate)-co-(1,4-butylene adipate)] (Ecoflex, BTA) monofilaments (rods) in standardized sandy soil was investigated. Changes in the microstructure and chemical composition distribution of the degraded BTA samples were evaluated and changes in the pH and salinity of postdegradation soil, as well as the soil phytotoxicity impact of the degradation products, are reported. A macroscopic and microscopic evaluation of the surface of BTA rod samples after specified periods of incubation in standardized soil indicated erosion of the surface of BTA rods starting from the fourth month of their incubation, with almost total disintegration of the incubated BTA material observed after 22 months. However, the weight loss after this period of time was about 50% and only a minor change in the M(w) of the investigated BTA samples was observed, along with a slight increase in the dispersity (from an initial 2.75 up to 4.00 after 22 months of sample incubation). The multidetector SEC and ESI-MS analysis indicated retention of aromatic chain fragments in the low molar mass fraction of the incubated sample. Phytotoxicity studies revealed no visible damage, such as necrosis and chlorosis, or other inhibitory effects, in the following plants: radish, cres, and monocotyledonous oat, indicating that the degradation products of the investigated BTA copolyester are harmless to the tested plants.

A new model integrating short- and long-term aging of copper added to soils

PubMed Central

Zeng, Saiqi; Li, Jumei; Wei, Dongpu

2017-01-01

Aging refers to the processes by which the bioavailability/toxicity, isotopic exchangeability, and extractability of metals added to soils decline overtime. We studied the characteristics of the aging process in copper (Cu) added to soils and the factors that affect this process. Then we developed a semi-mechanistic model to predict the lability of Cu during the aging process with descriptions of the diffusion process using complementary error function. In the previous studies, two semi-mechanistic models to separately predict short-term and long-term aging of Cu added to soils were developed with individual descriptions of the diffusion process. In the short-term model, the diffusion process was linearly related to the square root of incubation time (t1/2), and in the long-term model, the diffusion process was linearly related to the natural logarithm of incubation time (lnt). Both models could predict short-term or long-term aging processes separately, but could not predict the short- and long-term aging processes by one model. By analyzing and combining the two models, we found that the short- and long-term behaviors of the diffusion process could be described adequately using the complementary error function. The effect of temperature on the diffusion process was obtained in this model as well. The model can predict the aging process continuously based on four factors—soil pH, incubation time, soil organic matter content and temperature. PMID:28820888

Computational analysis of liquid chromatography-tandem mass spectrometric steroid profiling in NCI H295R cells following angiotensin II, forskolin and abiraterone treatment.

PubMed

Mangelis, Anastasios; Dieterich, Peter; Peitzsch, Mirko; Richter, Susan; Jühlen, Ramona; Hübner, Angela; Willenberg, Holger S; Deussen, Andreas; Lenders, Jacques W M; Eisenhofer, Graeme

2016-01-01

Adrenal steroid hormones, which regulate a plethora of physiological functions, are produced via tightly controlled pathways. Investigations of these pathways, based on experimental data, can be facilitated by computational modeling for calculations of metabolic rate alterations. We therefore used a model system, based on mass balance and mass reaction equations, to kinetically evaluate adrenal steroidogenesis in human adrenal cortex-derived NCI H295R cells. For this purpose a panel of 10 steroids was measured by liquid chromatographic-tandem mass spectrometry. Time-dependent changes in cell incubate concentrations of steroids - including cortisol, aldosterone, dehydroepiandrosterone and their precursors - were measured after incubation with angiotensin II, forskolin and abiraterone. Model parameters were estimated based on experimental data using weighted least square fitting. Time-dependent angiotensin II- and forskolin-induced changes were observed for incubate concentrations of precursor steroids with peaks that preceded maximal increases in aldosterone and cortisol. Inhibition of 17-alpha-hydroxylase/17,20-lyase with abiraterone resulted in increases in upstream precursor steroids and decreases in downstream products. Derived model parameters, including rate constants of enzymatic processes, appropriately quantified observed and expected changes in metabolic pathways at multiple conversion steps. Our data demonstrate limitations of single time point measurements and the importance of assessing pathway dynamics in studies of adrenal cortical cell line steroidogenesis. Our analysis provides a framework for evaluation of steroidogenesis in adrenal cortical cell culture systems and demonstrates that computational modeling-derived estimates of kinetic parameters are an effective tool for describing perturbations in associated metabolic pathways. Copyright © 2015 Elsevier Ltd. All rights reserved.

Nematicidal activity of three novel extracellular proteases of the nematophagous fungus Monacrosporium sinense.

PubMed

Soares, Filippe E F; Braga, Fabio R; Araújo, Jackson V; Geniêr, Hugo L A; Gouveia, Angélica S; Queiroz, José H

2013-04-01

Extracellular proteases are an important virulence factor for the nematophagous fungi Monacrosporium. The objective of this study was to optimize, purify, partially characterize, and to evaluate the nematicidal activity of the proteases produced by the nematophagous fungus Monacrosporium sinense (SF53) by solid-state fermentation. Wheat bran was used as substrate for protease production. The variables moisture, pH, incubation time, temperature, glucose, yeast extract, and the number of conidia were tested for their influences on protease production by SF53. To determine the optimal level of the selected variables the central composite design was applied. The crude extract obtained was purified in two steps, an ion exchange chromatography and a gel excision. SDS-PAGE and zymogram were performed for analysis of the purification process. Proteolytic activity was also tested at different pHs and temperatures. In the in vitro assay, the nematicidal activity of the three proteases was evaluated. pH and incubation time showed a significant effect (p<0.05) on production of protease. The highest value of activity was 38.0 (U/ml) under the conditions of pH 5.0 and incubation time of 211 h. SF53 produced three different proteases (Ms1, Ms2, and Ms3) which were directly purified from the zymogram. Ms1, Ms2, and Ms3 showed the following percentage of reduction (p<0.05) on the number of Panagrellus redivivus compared to control after 24 h: 76.8, 68.1, and 92.1%. This is the first report of the use of proteases of the isolate SF53 on a phytonematode, which may be a research tool in future works.

Copper stimulation of LHRH release from median eminence explants. III. A process dependent on extracellular sodium.

PubMed

Colombani-Vidal, M; Barnea, A

1986-01-01

Copper, complexed to histidine (CuHis), stimulates LHRH release from explants of the median eminence area (MEA). To gain further understanding of the mechanism of copper action, in this study, we assessed the Na+ and energy requirements for CuHis stimulation of LHRH release. MEA explants, obtained from adult male rats, were incubated at 37 degrees C for 15 min with 100 microM CuHis and then for 45 min in CuHis-free medium (Krebs-Ringer-phosphate buffer, pH 7.4). LHRH released into the medium was evaluated by RIA. When the incubation buffer contained 143 mM Na+, CuHis stimulated the release of LHRH from a basal level of 17.2 +/- 1.26 (mean +/- SEM, n = 7) to 74.5 +/- 6.2 pg/60 min per MEA. When [Na+] was reduced to 16 mM Na+ (by substituting with Li+), CuHis-stimulated LHRH release was inhibited by 80% (p less than 0.001); indicating a requirement for Na+. In addition, we found that CuHis-stimulated LHRH release was a saturable function of Na+ concentration; saturation achieved with about 100 mM Na+. To assess the requirement for Na+ transport, we evaluated the effect of 1 mM ouabain, 10 microM tetrodotoxin (TTX), or 100 microM amiloride on CuHis stimulation of LHRH release. Ouabain inhibited CuHis stimulation of LHRH release by 80%, whereas TTX and amiloride were ineffective. In addition, we observed that CuHis did not stimulate LHRH release when incubation was carried out at 4 degrees C or at 37 degrees C in the presence of 5 mM KCN.(ABSTRACT TRUNCATED AT 250 WORDS)

A membrane-associated adenylate cyclase modulates lactate dehydrogenase and creatine kinase activities required for bull sperm capacitation induced by hyaluronic acid.

PubMed

Fernández, Silvina; Córdoba, Mariana

2017-04-01

Hyaluronic acid, as well as heparin, is a glycosaminoglycan present in the female genital tract of cattle. The aim of this study was to evaluate oxidative metabolism and intracellular signals mediated by a membrane-associated adenylate cyclase (mAC), in sperm capacitation with hyaluronic acid and heparin, in cryopreserved bull sperm. The mAC inhibitor, 2',5'-dideoxyadenosine, was used in the present study. Lactate dehydrogenase (LDH) and creatine kinase (CK) activities and lactate concentration were determined spectrophotometrically in the incubation medium. Capacitation and acrosome reaction were evaluated by chlortetracycline technique, while plasma membrane and acrosome integrity were determined by trypan blue stain/differential interference contrast microscopy. Heparin capacitated samples had a significant decrease in LDH and CK activities, while in hyaluronic acid capacitated samples LDH and CK activities both increased compared to control samples, in heparin and hyaluronic acid capacitation conditions, respectively. A significant increase in lactate concentration in the incubation medium occurred in hyaluronic acid-treated sperm samples compared to heparin treatment, indicating this energetic metabolite is produced during capacitation. The LDH and CK enzyme activities and lactate concentrations in the incubation medium were decreased with 2',5'-dideoxyadenosine treatment in hyaluronic acid samples. The mAC inhibitor significantly inhibited heparin-induced capacitation of sperm cells, but did not completely inhibit hyaluronic acid capacitation. Therefore, hyaluronic acid and heparin are physiological glycosaminoglycans capable of inducing in vitro capacitation in cryopreserved bull sperm, stimulating different enzymatic pathways and intracellular signals modulated by a mAC. Hyaluronic acid induces sperm capacitation involving LDH and CK activities, thereby reducing oxidative metabolism, and this process is mediated by mAC. Copyright © 2017 Elsevier B.V. All rights reserved.

Evaluation of a PCR assay on overgrown environmental samples cultured for Mycobacterium avium subsp. paratuberculosis.

PubMed

Arango-Sabogal, Juan C; Labrecque, Olivia; Paré, Julie; Fairbrother, Julie-Hélène; Roy, Jean-Philippe; Wellemans, Vincent; Fecteau, Gilles

2016-11-01

Culture of Mycobacterium avium subsp. paratuberculosis (MAP) is the definitive antemortem test method for paratuberculosis. Microbial overgrowth is a challenge for MAP culture, as it complicates, delays, and increases the cost of the process. Additionally, herd status determination is impeded when noninterpretable (NI) results are obtained. The performance of PCR is comparable to fecal culture, thus it may be a complementary detection tool to classify NI samples. Our study aimed to determine if MAP DNA can be identified by PCR performed on NI environmental samples and to evaluate the performance of PCR before and after the culture of these samples in liquid media. A total of 154 environmental samples (62 NI, 62 negative, and 30 positive) were analyzed by PCR before being incubated in an automated system. Growth was confirmed by acid-fast bacilli stain and then the same PCR method was again applied on incubated samples, regardless of culture and stain results. Change in MAP DNA after incubation was assessed by converting the PCR quantification cycle (Cq) values into fold change using the 2 -ΔCq method (ΔCq = Cq after culture - Cq before culture). A total of 1.6% (standard error [SE] = 1.6) of the NI environmental samples had detectable MAP DNA. The PCR had a significantly better performance when applied after culture than before culture (p = 0.004). After culture, a 66-fold change (SE = 17.1) in MAP DNA was observed on average. Performing a PCR on NI samples improves MAP culturing. The PCR method used in our study is a reliable and consistent method to classify NI environmental samples. © 2016 The Author(s).

Hypogravity's Effect on the Life Cycle of Japanese Quail

NASA Technical Reports Server (NTRS)

Hester, Patricia Y.

1999-01-01

A series of studies were conducted to determine the effect of activities preceding space-flight and during space-flight on quail embryonic development. While the overall development of the quail embryos was evaluated, the report presented herein, focused on calcium utilization or uptake from eggshells by developing embryos during incubation in space and on earth. In the pre-space trials, fertilized quail eggs were subjected to pre-night dynamics including forces of centrifugation, vibration, or a combination of vibration and centrifugation prior to incubation for 6 or 16 days. In another trial, fertile quail eggs were tested for survivability in a refrigerator stowage kit for eggs (RSKE) which was subsequently used to transport the eggs to space. Eggs in the RSKE were subjected to shuttle launch dynamics including G force and random vibration profiles. In the space- flight trials, 48 fertile quail eggs were launched on space shuttle Flight STS-76 and were subsequently incubated in a Slovakian incubator onboard space station, MIR. Two sets of ground controls each with 48 fertile eggs with and without exposure to launch dynamics were initiated 5 days post-launch. There was a laboratory control (incubated in Lyon RX2 incubator at 37.5 C) and a synchronous control (incubated in Lyon RX2 incubator at 39 - 400 C), which simulated the temperature of the space-flight incubator. Following space-flight trials, post-flight trials were conducted where quail eggs were incubated in Lyon RX2 or Slovakian incubators under various temperatures with or without launch dynamics. Eggshells from all study trials were retrieved and analyzed for calcium content to determine if its utilization by developing quail embryos was affected by activities preceding space-flight or during incubation in space under microgravity. Results from the pre-flight and post-flight showed that pre-flight activities and shuttle launch dynamics had no effect on calcium uptake from the eggshell by developing embryos. However, calcium uptake from the eggshell by developing embryos incubated in micro,aravity was impaired by 12.6% when compared to embryos incubated on earth under laboratory control environment. This impairment was unlikely due to factors other than microgravity. In general, calcium utilization by developing embryos increased with age of incubation with the most increase occurring at day 16 of incubation.

A model to predict the thermal reaction norm for the embryo growth rate from field data.

PubMed

Girondot, Marc; Kaska, Yakup

2014-10-01

The incubation of eggs is strongly influenced by temperature as observed in all species studied to date. For example, incubation duration, sexual phenotype, growth, and performances in many vertebrate hatchlings are affected by incubation temperature. Yet it is very difficult to predict temperature effect based on the temperature within a field nest, as temperature varies throughout incubation. Previous works used egg incubation at constant temperatures in the laboratory to evaluate the dependency of growtProd. Type: FTPh rate on temperature. However, generating such data is time consuming and not always feasible due to logistical and legislative constraints. This paper therefore presents a methodology to extract the thermal reaction norm for the embryo growth rate directly from a time series of incubation temperatures recorded within natural nests. This methodology was successfully applied to the nests of the marine turtle Caretta caretta incubated on Dalyan Beach in Turkey, although it can also be used for any egg-laying species, with some of its limitations being discussed in the paper. Knowledge about embryo growth patterns is also important when determining the thermosensitive period for species with temperature-dependent sex determination. Indeed, in this case, sexual phenotype is sensitive to temperature only during this window of embryonic development. Copyright © 2014 Elsevier Ltd. All rights reserved.

Screening of agro-industrial wastes for citric acid bioproduction by Aspergillus niger NRRL 2001 through solid state fermentation.

PubMed

Dhillon, Gurpreet S; Brar, Satinder K; Kaur, Surinder; Verma, Mausam

2013-05-01

The citric acid (CA) industry is currently struggling to develop a sustainable and economical process owing to high substrate and energy costs. Increasing interest in the replacement of costly synthetic substrates by renewable waste biomass has fostered research on agro-industrial wastes and screening of raw materials for economical CA production. The food-processing industry generates substantial quantities of waste biomass that could be used as a valuable low-cost fermentation substrate. The present study evaluated the potential of different agro-industrial wastes, namely apple pomace (AP), brewer's spent grain, citrus waste and sphagnum peat moss, as substrates for solid state CA production using Aspergillus niger NRRL 2001. Among the four substrates, AP resulted in highest CA production of 61.06 ± 1.9 g kg(-1) dry substrate (DS) after a 72 h incubation period. Based on the screening studies, AP was selected for optimisation studies through response surface methodology (RSM). Maximum CA production of 312.32 g kg(-1) DS was achieved at 75% (v/w) moisture and 3% (v/w) methanol after a 144 h incubation period. The validation of RSM-optimised parameters in plastic trays resulted in maximum CA production of 364.4 ± 4.50 g kg(-1) DS after a 120 h incubation period. The study demonstrated the potential of AP as a cheap substrate for higher CA production. This study contributes to knowledge about the future application of carbon rich agro-industrial wastes for their value addition to CA. It also offers economic and environmental benefits over traditional ways used to dispose off agro-industrial wastes. © 2012 Society of Chemical Industry.

Nitrogen availability from residues-based biochar at two pyrolisis temperatures

NASA Astrophysics Data System (ADS)

Coscione, Aline Renee; Silveira Bibar, Maria Paula; de Andrade, Cristiano Alberto

2014-05-01

Biochar has been studied for several applications, such as soil quality improvement, heavy metals remediation and N2O mitigation. Considering the soil quality improvement aspect it is desirable to evaluate if the nitrogen content in biochar samples obtained from several residues used as the biomass sources could be available for plants. Samples of sewage sludge (SS), coffee grounds (CG), chicken manure (CM) and fungi mycelia (FM) were pyrolyzed at two temperatures, 400 and 700 oC (indicated by the number 4 and 7 in this abstract, respectively), in order to obtain the biochar samples. The Kjeldahl nitrogen of biochar was (% m/m): 3.0 (CM4, CG7, FM7 and CG4); 2.0 (CM7 e SS4); 3.4 (FM7); 1.4 (SS7), with organic carbon (potassium dichromate method) ranging from 2.0 to 3.0% for all but CG4 (6%). The C/N ratio of biochar samples was: 9 (CM4, SS4 and CG7); 11 (CM7); 15 (SS7); 7 (FM4 and FM7); 21 (CG4). The eight soil + biochar resulting mixtures, prepared using the equivalent to 60 t/ha of biochar (about 3% w/w), and one additional control treatment (no biochar added) were incubated for 90 days, with four replications of each treatment per time evaluated. Inorganic nitrogen and soil pH measurements were performed for all treatments at 0, 5, 15, 30, 60 and 90 days of incubation. Soil moisture was kept at 40% soil water holding capacity, by weighting, during the experiment. The data was submitted to ANOVA with Tukey's average comparison test (p < 0.05). No significative pH changes were observed during the incubation of biochar samples. At the initial incubation time (zero days) no statistical difference was observed among biochar sources or pyrolisis temperatures. After five days of incubation SS4 and CM4 showed significant inorganic nitrogen release compared to all other treatments, behavior repeated at all the following times evaluated. For CM7, FM4 and FM7 maximum nitrogen availability was observed after 15 days, while it occurred after 90 days for SS4. After 90 days, only SS4 and CM4 presented a positive nitrogen balance, reaching 8 and 9 % of the nitrogen added by biochar samples release to the soil, respectively. A first order kinetic model was adjusted for SS4 nitrogen release, enabling the calculation of half life (10 days), potential available nitrogen (76.5 mg/kg) and the speed of the process. However, compared to SS4 the standard nitrogen availability of sewage sludge is up to 30% of its Kjeldahl nitrogen. For organic residues with C/N ratios lower than 20 applied to the soil a fast degradation, with the corresponding increase in inorganic nitrogen availability is expect. Although all the biochar samples tested had C/N ratios below that cutting point, just 2 of 8 presented inorganic nitrogen available in the soil+biochar mixtures. These results show that soil incubation tests are ultimate for the evaluation of the nitrogen potential release to the soil. Low temperature SS based biochar may offer additional nitrogen release to soil besides other soil conditioning properties.

Effects of human recreation on the incubation behavior of American Oystercatchers

USGS Publications Warehouse

McGowan, C.P.; Simons, T.R.

2006-01-01

Human recreational disturbance and its effects on wildlife demographics and behavior is an increasingly important area of research. We monitored the nesting success of American Oystercatchers (Haematopus palliatus) in coastal North Carolina in 2002 and 2003. We also used video monitoring at nests to measure the response of incubating birds to human recreation. We counted the number of trips per hour made by adult birds to and from the nest, and we calculated the percent time that adults spent incubating. We asked whether human recreational activities (truck, all-terrain vehicle [ATV], and pedestrian traffic) were correlated with parental behavioral patterns. Eleven a priori models of nest survival and behavioral covariates were evaluated using Akaike's Information Criterion (AIC) to see whether incubation behavior influenced nest survival. Factors associated with birds leaving their nests (n = 548) included ATV traffic (25%), truck traffic (17%), pedestrian traffic (4%), aggression with neighboring oystercatchers or paired birds exchanging incubation duties (26%), airplane traffic (1%) and unknown factors (29%). ATV traffic was positively associated with the rate of trips to and away from the nest (??1 = 0.749, P < 0.001) and negatively correlated with percent time spent incubating (??1 = -0.037, P = 0.025). Other forms of human recreation apparently had little effect on incubation behaviors. Nest survival models incorporating the frequency of trips by adults to and from the nest, and the percentage of time adults spent incubating, were somewhat supported in the AIC analyses. A low frequency of trips to and from the nest and, counter to expectations, low percent time spent incubating were associated with higher daily nest survival rates. These data suggest that changes in incubation behavior might be one mechanism by which human recreation affects the reproductive success of American Oystercatchers.

[Influence of incubation time on metabolites in mycelia of Paecilomyces militaris].

PubMed

Zhang, Delong; Li, Shulin; Lu, Ruili; Li, Kangle; Luo, Feifei; Peng, Fan; Hu, Fenglin

2012-12-04

To determine the secondary metabolites production in mycelia of Paecilomyces militaris. Mycelia were cultured in plates with sabouraud dextrose agar yeast medium at 25 degrees C for 9 days. Sampling was done every day from the second to the ninth day. The secondary metabolites in the mycelia of Paecilomyces militaris were extracted with either methanol or ethyl acetate. The extracts were blended and analyzed by liquid chromatography-mass spectrometry (LC-MS). LC-MS data were collected and analyzed by MetaboAnalyst software. Principal component analysis indicates different secondary metabolites accumulation with incubation times. Hierarchical clustering analysis shows that the metabolic process of cationic compounds such as alkaloids, peptides and nucleosides can be divided into three stages, and that the metabolic process of anionic compounds such as organic acids and saccharides can be divided into two stages. Metabolites difference and heat map analysis show that: (1) The number of metabolites with significant increased contents was raised significantly in mycelia of Paecilomyces militaris on the second and third incubation days. The main species with increased contents were esters and their hydrolized products, destruxin B, variotin and some unidentified nitrogin contained compounds. (2) The number of metabolites with significant raised contents was decreased significantly on the fourth and fifth incubation days. The main species with increased contents were ophiocordin and destruxin A. (3) Apart from peptide antibiotics such as several beauverolides, the content increased metabolites included also several organic acids, amino acids, rhamnose, trehalose, cerebroside and riboflavine during the sixth to ninth incubation days. The secondary metabolites in mycelia of Paecilomyces militaris were related significantly to the incubation time.

Fucoidan as an inhibitor of thermally induced collagen glycation examined by acetate electrophoresis.

PubMed

Pielesz, Anna; Paluch, Jadwiga

2014-08-01

Non-enzymatic glycation (Maillard reaction) in vitro could be a simple method to obtain glycoconjugates for studying their biological properties. Hence, fucoidan was retained by acetate electrophoresis indicating a strong interaction with the protein. A loss of colour in fucoidan bands was found for samples incubated with collagen as compared with samples of free fucoidan. Also under in vitro conditions at 100°C - simulating a sudden burn incident - fucoidan binds with collagen as a result of the Maillard reaction. In contrast, the colour of the fucoidan bands intensified for samples incubated with collagen, with the addition of glucose. Electrophoretic analyses were carried out after heating the samples to a temperature simulating a burn incident. The bands were found to intensify for samples incubated with collagen during a 30-day-long incubation. Thus, spontaneous in vitro glycation - i.e. without the addition of glucose - was confirmed. This process is highly intensified both by the temperature and time of incubation. For a sample incubated in vitro in a fucoidan solution containing glucose, glycation was confirmed in a preliminary FTIR and acetate electrophoresis examinations, occurring in collagen obtained from chicken skins. In particular, a new band emerging around 1746 cm(-1) was observed for above samples, as was its increasing intensity, as compared with samples without the addition of glucose. In the collagen glycation assay, while glucose reacts with collagen and forms cross-linked aggregates, fucoidan decreases the process of aggregation and recovery of native collagen. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Qualification of a rapid readout biological indicator with moist heat sterilization.

PubMed

McCormick, Patrick; Finocchario, Catherine; Manchester, Robert; Glasgow, Louis; Costanzo, Stephen

2003-01-01

Biological indicators are recognized as an important component in the validation and routine monitoring of moist heat (steam) sterilization processes. Due to the need to allow for the recovery and outgrowth of test organisms that may have been sub-lethally injured, between 2-5 days of incubation are typically required before the outcome of sterilization processing can be reliably interpreted. Rapid readout biological indicators that incorporate the response of a heat resistant enzyme provide a means for assessing the efficacy of moist heat sterilization within hours of processing. This study describes the qualification of the 3M Attest 1292 Rapid Readout Biological Indicator with moist heat sterilization according to the procedures described in the PDA Technical Report No. 33, "Evaluation, Validation and Implementation of New Microbiological Testing Methods".

Chick embryogenesis: a unique platform to study the effects of environmental factors on embryo development.

PubMed

Yahav, S; Brake, J

2014-01-01

Bird embryogenesis takes place in a relatively protected environment that can be manipulated especially well in domestic fowl (chickens) where incubation has long been a commercial process. The embryonic developmental process has been shown to begin in the oviduct such that the embryo has attained either the blastodermal and/or gastrulation stage of development at oviposition. Bird embryos can be affected by "maternal effects," and by environmental conditions during the pre-incubation and incubation periods. "Maternal effects" has been described as an evolutionary mechanism that has provided the mother, by hormonal deposition into the yolk, with the potential to proactively influence the development of her progeny by exposing them to her particular hormonal pattern in such a manner as to influence their ability to cope with the expected wide range of environmental conditions that may occur post-hatching. Another important aspect of "maternal effects" is the effect of the maternal nutrient intake on progeny traits. From a commercial broiler chicken production perspective, it has been established that greater cumulative nutrient intake by the hen during her pullet rearing phase prior to photostimulation resulted in faster growing broiler progeny. Generally, maternal effects on progeny, which have both a genetic and an environmental component represented by yolk hormones deposition and embryo nutrient utilization, have an important effect on the development of a wide range of progeny traits. Furthermore, commercial embryo development during pre-incubation storage and incubation, as well as during incubation per se has been shown to largely depend upon temperature, while other environmental factors that include egg position during storage, and the amount of H2O and CO2 lost by the egg and the subsequent effect on albumen pH and height during storage have become important environmental factors to be considered for successful embryogenesis under commercial conditions. Manipulating environmental temperature during the period of egg storage, during the intermediate pre-incubation period, and incubation period per se has been found to significantly affect embryo development, hatching progress, chick quality at hatching, and chick development post-hatching. These temperature manipulations have also been shown to affect the acquisition of thermotolerance to subsequent post-hatching thermal challenge. This chapter will focus on: a. "maternal effects" on embryo and post-hatching development; b. environmental effects during the post-ovipositional period of egg storage, the intermediate pre-incubation period, and incubation period per se on chick embryogenesis and subsequent post-hatching growth and development; and c. effects of temperature manipulations during the pre-incubation and incubation periods on acquisition of thermotolerance and development of secondary sexual characteristics in broiler chickens.

A Meta-Analysis of Experiments Linking Incubation Conditions with Subsequent Leg Weakness in Broiler Chickens

PubMed Central

Groves, Peter J.; Muir, Wendy I.

2014-01-01

A series of incubation and broiler growth studies were conducted using one strain of broiler chicken (fast feathering dam line) observing incubation effects on femoral bone ash % at hatch and the ability of the bird to remain standing at 6 weeks of age (Latency-To-Lie). Egg shell temperatures during incubation were consistently recorded. Parsimonious models were developed across eight studies using stepwise multiple linear regression of egg shell temperatures over 3-day periods and both bone ash at hatch and Latency-To-Lie. A model for bone ash at hatch explained 70% of the variation in this factor and revealed an association with lower egg shell temperatures during days 4–6 and 13–15 and higher egg shell temperatures during days 16–18 of incubation. Bone ash at hatch and subsequent Latency-To-Lie were positively correlated (r = 0.57, P<0.05). A model described 66% of the variation Latency-To-Lie showing significant association of the interaction of femoral ash at hatch and lower average egg shell temperatures over the first 15 days of incubation. Lower egg shell temperature in the early to mid incubation process (days 1–15) and higher egg shell temperatures at a later stage (days 16–18) will both tend to delay the hatch time of incubating eggs. Incubation profiles that resulted in later hatching chicks produced birds which could remain standing for a longer time at 6 weeks of age. This supports a contention that the effects of incubation observed in many studies may in fact relate more to earlier hatching and longer sojourn of the hatched chick in the final stage incubator. The implication of these outcomes are that the optimum egg shell temperature during incubation for broiler leg strength development may be lower than that regarded as ideal (37.8°C) for maximum hatchability and chick growth. PMID:25054636

A meta-analysis of experiments linking incubation conditions with subsequent leg weakness in broiler chickens.

PubMed

Groves, Peter J; Muir, Wendy I

2014-01-01

A series of incubation and broiler growth studies were conducted using one strain of broiler chicken (fast feathering dam line) observing incubation effects on femoral bone ash % at hatch and the ability of the bird to remain standing at 6 weeks of age (Latency-To-Lie). Egg shell temperatures during incubation were consistently recorded. Parsimonious models were developed across eight studies using stepwise multiple linear regression of egg shell temperatures over 3-day periods and both bone ash at hatch and Latency-To-Lie. A model for bone ash at hatch explained 70% of the variation in this factor and revealed an association with lower egg shell temperatures during days 4-6 and 13-15 and higher egg shell temperatures during days 16-18 of incubation. Bone ash at hatch and subsequent Latency-To-Lie were positively correlated (r = 0.57, P<0.05). A model described 66% of the variation Latency-To-Lie showing significant association of the interaction of femoral ash at hatch and lower average egg shell temperatures over the first 15 days of incubation. Lower egg shell temperature in the early to mid incubation process (days 1-15) and higher egg shell temperatures at a later stage (days 16-18) will both tend to delay the hatch time of incubating eggs. Incubation profiles that resulted in later hatching chicks produced birds which could remain standing for a longer time at 6 weeks of age. This supports a contention that the effects of incubation observed in many studies may in fact relate more to earlier hatching and longer sojourn of the hatched chick in the final stage incubator. The implication of these outcomes are that the optimum egg shell temperature during incubation for broiler leg strength development may be lower than that regarded as ideal (37.8°C) for maximum hatchability and chick growth.

Gelation of high-methoxy pectin by enzymic de-esterification in the presence of calcium ions: a preliminary evaluation.

PubMed

O'Brien, Aileen B; Philp, Kevin; Morris, Edwin R

2009-09-28

Cohesive gels have been obtained by de-esterification of 1.0wt% high-methoxy citrus pectin (degree of esterification approximately 68%) in the presence of Ca(2+) cations, using a commercial preparation (NovoShape) of fungal methyl esterase cloned from Aspergillus aculeatus. A convenient rate of network formation (gelation within approximately 30min) was achieved at an enzyme concentration of 0.2 PEU/g pectin. At a Ca(2+)-concentration of 40mM and incubation temperature of 20 degrees C, severe syneresis (>7% of sample mass) was observed, but release of fluid decreased with decreasing concentration of Ca(2+) and increasing temperature of incubation, becoming undetectable for 10mM Ca(2+) at 30 degrees C. Under these conditions, progressive development of solid-like character (storage modulus, G') was observed during 160min of enzymic de-esterification, and the mechanical spectrum recorded at the end of the incubation period had the form typical of a biopolymer gel. On subsequent heating to 70 degrees C, dissociation of the gel network (sigmoidal reduction in G' and G'') was observed. At or above the midpoint temperature of this melting process ( approximately 50 degrees C), there was no indication of gel formation on enzymic de-esterification (at 50 or 60 degrees C). At lower temperatures (20, 30 and 40 degrees C), the rate of gelation (assessed visually) showed no systematic increase as the incubation temperature was increased towards the temperature-optimum of the enzyme ( approximately 50 degrees C). This unexpected behaviour is attributed to competition between faster de-esterification and slower formation of Ca(2+)-induced 'egg-box' junctions.

Effect of volatile fatty acids in anaerobic conditions on viability of helminth ova (Ascaris suum) in sanitization of municipal sludge.

PubMed

Rojas-Oropeza, Marcelo; Hernández-Uresti, Alejandro S; Ortega-Charleston, Luis S; Cabirol, Nathalie

2017-09-01

The present work aimed at evaluating the effect of four different mixtures of diverse volatile fatty acids (VFAs) on the viability of helminth ova (Ascaris suum), under mesophilic (35°C) anaerobic conditions and at different incubation times, in order to reproduce the process of two-phase anaerobic digestion. The mixtures of VFAs contained acetic, propionic, butyric, valeric, and isovaleric acids, used at concentrations normally found in acidogenic anaerobic digesters. The four treatments all showed a reduction in Ascaris suum ova viability, among which Treatment III (4.2 g-acetic acid L -1  +  2.2 g-propionic acid L -1  + 0.6 g-valeric acid L -1  + 0.6 g-isovaleric acid L -1 ) resulted the most efficient. We found that the full effect of VFAs on the viability loss of Ascaris suum ova in mesophilic conditions requires a minimum incubation time of 3 days. The highest efficiency in the loss of viability was observed with Treatment III and 4-day incubation. Interestingly, the proportion of acetic acid was three times as much in this treatment than in the other ones and resulted in an effect in a minimum time of 3 days. The mesophilic condition, however, was not sufficient to induce a complete loss of viability.

Evaluation of petrifilm series 2000 as a possible rapid method to count coliforms in foods.

PubMed

Priego, R; Medina, L M; Jordano, R

2000-08-01

This research note is a preliminary comparison between the Petrifilm 2000 method and a widely used traditional enumeration method (on violet red bile agar); six batches of different foods (egg, frozen green beans, fresh sausage, a bakery product, raw minced meat, and raw milk) were studied. The reliability of the presumptive counts taken at 10, 12, and 14 h of incubation using this method was also verified by comparing the counts with the total confirmed counts at 24 h. In all the batches studied, results obtained with Petrifilm 2000 presented a close correlation to those obtained using violet red bile agar (r = 0.860) and greater sensitivity (93.33% of the samples displayed higher counts on Petrifilm 2000), showing that this method is a reliable and efficient alternative. The count taken at 10-h incubation is of clear interest as an early indicator of results in microbiological food control, since it accounted for 90% of the final count in all the batches analyzed. Counts taken at 12 and 14 h bore a greater similarity to those taken at 24 h. The Petrifilm 2000 method provides results in less than 12 h of incubation, making it a possible rapid method that adapts perfectly to hazard analysis critical control point system by enabling the microbiological quality control of the processing.

Effects of Environmental Factors on Soluble Expression of a Humanized Anti-TNF-α scFv Antibody in Escherichia coli

PubMed Central

Sina, Mohammad; Farajzadeh, Davoud; Dastmalchi, Siavoush

2015-01-01

Purpose: The bacterial cultivation conditions for obtaining anti-TNF-α single chain variable fragment (scFv) antibody as the soluble product in E. coli was investigated. Methods: To avoid the production of inclusion bodies, the effects of lactose, IPTG, incubation time, temperature, shaking protocol, medium additives (Mg+2, sucrose), pH, osmotic and heat shocks were examined. Samples from bacterial growth conditions with promising results of soluble expression of GST-hD2 scFv were affinity purified and quantified by SDS-PAGE and image processing for further evaluation. Results: The results showed that cultivation in LB medium under induction by low concentrations of lactose and incubation at 10 °C led to partial solubilization of the expressed anti-TNF-α scFv (GST-hD2). Other variables which showed promising increase in soluble expression of GST-hD2 were osmotic shock and addition of magnesium chloride. Furthermore, addition of sucrose to medium suppressed the expression of scFv completely. The other finding was that the addition of sorbitol decreased the growth rate of bacteria. Conclusion: It can be concluded that low cultivation temperature in the presence of low amount of inducer under a long incubation time or addition of magnesium chloride are the most effective environmental factors studied for obtaining the maximum solubilization of GST-hD2 recombinant protein. PMID:26819916

Effects of Environmental Factors on Soluble Expression of a Humanized Anti-TNF-α scFv Antibody in Escherichia coli.

PubMed

Sina, Mohammad; Farajzadeh, Davoud; Dastmalchi, Siavoush

2015-11-01

The bacterial cultivation conditions for obtaining anti-TNF-α single chain variable fragment (scFv) antibody as the soluble product in E. coli was investigated. To avoid the production of inclusion bodies, the effects of lactose, IPTG, incubation time, temperature, shaking protocol, medium additives (Mg+2, sucrose), pH, osmotic and heat shocks were examined. Samples from bacterial growth conditions with promising results of soluble expression of GST-hD2 scFv were affinity purified and quantified by SDS-PAGE and image processing for further evaluation. The results showed that cultivation in LB medium under induction by low concentrations of lactose and incubation at 10 °C led to partial solubilization of the expressed anti-TNF-α scFv (GST-hD2). Other variables which showed promising increase in soluble expression of GST-hD2 were osmotic shock and addition of magnesium chloride. Furthermore, addition of sucrose to medium suppressed the expression of scFv completely. The other finding was that the addition of sorbitol decreased the growth rate of bacteria. It can be concluded that low cultivation temperature in the presence of low amount of inducer under a long incubation time or addition of magnesium chloride are the most effective environmental factors studied for obtaining the maximum solubilization of GST-hD2 recombinant protein.

Evaluation of rapid readout biological indicators for 132 degrees C gravity and 132 degrees C vacuum-assisted steam sterilization cycles using a new automated fluorescent reader.

PubMed

Alfa, Michelle J; Olson, Nancy; DeGagne, Pat; Jackson, Michele

2002-07-01

The primary objective of this study was to evaluate fluorescent readout results of Attest 1291 Biological Indicators (BIs) (3M Health Care, St. Paul, MN) and Attest 1296 BI test packs (containing Attest 1292 BIs) using full and fractional cycles compared with the growth data when prolonged incubation (7 days) was included. Gravity displacement and vacuum-assisted steam sterilization cycles were evaluated. A secondary objective of this study was to evaluate the new automated rapid fluorescent reader (Attest 290 Auto Reader). The rapid readout BIs for gravity displacement and vacuum-assisted steam autoclave cycles at 132 degrees C were processed using full (4 minutes) and four fractional cycles that provided 30% to 80% positive results for growth after 24 hours of incubation (48 hours of incubation for Attest 1292 BIs from the Attest 1296 test packs). Sixty of each type of BI were tested for each cycle (300 of each BI type in total). For all full steam sterilization cycles, results of the rapid fluorescent readout and the 24-hour, 48-hour, and 7-day growth tests were negative for all Attest 1291 and 1292 BIs tested. For all fractional cycles, the 24- and 48-hour growth results for the Attest 1291 and 1292 BIs, respectively, were the same as the 7-day growth results. The fractional cycle data indicated that fluorescent rapid readout was a more sensitive indicator than growth. There were rare (0.9%) false-negative results for BIs under fractional cycle conditions and these all correlated with short fractional cycle exposure times. The fluorescent rapid readout results of the 1291 BIs and 1296 BI test packs reliably predict both 24- and 48-hour and 7-day growth. These data support the value of rapid readout BIs for sterilizer monitoring for both the vacuum-assisted and the gravity displacement steam sterilization cycles. The new automated reader requires less manipulation of the BI and makes monitoring user friendly and less prone to user errors.

Release of native and amended boron from arid zone soils after varying incubation times

USDA-ARS?s Scientific Manuscript database

In this study we evaluated the boron (B) release from soils containing elevated native B and examined the extent to which incubation time affected B release. Five soils varying in initial pH and clay content were selected for the study. The soils were spiked with five varying doses of B as H3BO3 (...

In an Egg Shell: Egg to Chick to Egg.

ERIC Educational Resources Information Center

Lyon Electric Company, Chula Vista, CA.

The goals of this program include enabling students to learn about the anatomy of an avian egg, egg formation, bird embryo development, and the process of egg incubation. This guide is designed to accompany the hands-on experience of incubation and hatching chicken eggs and is organized in three sections. The teaching materials section includes…

Acoustic pollution in hospital environments

NASA Astrophysics Data System (ADS)

Olivera, J. M.; Rocha, L. A.; Rotger, V. I.; Herrera, M. C.

2011-12-01

There are many different services within a hospital. This means different types of noise which can be considered as acoustic pollution. Knowing that preterm infants exposed to high amounts of noise in the NICU are at a much higher risk because of their neurologic immaturity and physiologic instability, that excessive levels of noise also affect the persons and it can also impede some studies on patients, it was proposed to evaluate the Sound Pressure Level in some services of the Instituto de Maternidad, Tucumán, Argentina. There were evaluated the Level III NICU, the laundry service, a physical space destined for a service of evoked potential and a neonatal incubator under working conditions. The measurements were performed with a type II sonometer (CENTER 322) and it was also used an incubator analyzer (FLUKE INCU) for the incubator. The average values obtained were of 63.6 dBA for the NICU, 82.5dBA for the laundry room, 52.7 dBA for the evoked potential room and 62.8 dBA in the inside of the incubator under 64 dBA in the outside. The reports were documented in compliance with the appropriate standards.

Newborn physiological responses to noise in the neonatal unit.

PubMed

Cardoso, Sandra Maria Schefer; Kozlowski, Lorena de Cássia; Lacerda, Adriana Bender Moreira de; Marques, Jair Mendes; Ribas, Angela

2015-01-01

The incorporation of technologies in the care of infants has contributed to increased survival; however, this has turned neonatal unit into a noisy environment. To evaluate the physiological and functional effects resulting from the exposure to noise on low-weight newborns in incubators in a neonatal unit. Prospective, observational, quantitative, exploratory, descriptive study. The adopted statistical method included tables of frequency, descriptive statistics, and Student's t-test, with a 0.05 level of significance. As data collection tools, the environmental noise and the noise inside of the incubator were evaluated, and the Assessment of Preterm Infant Behavior scale was used to assess premature newborn behavior and projected specifically to document the neurobehavioral functioning of preterm infants. The data collection occurred from September of 2012 to April of 2013; 61 low-weight newborns admitted in the neonatal unit and in incubators were observed. Significant differences in the variables heart rate and oxygen saturation were noted when newborns were exposed to noise. Low-weight neonates in incubators present physiological alterations when facing discomfort caused by environmental noise in neonatal units. Copyright © 2015 Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial. Published by Elsevier Editora Ltda. All rights reserved.

Surface Interactions between Escherichia coli and Hemocytes of the Mediterranean Mussel Mytilus galloprovincialis Lam. Leading to Efficient Bacterial Clearance

PubMed Central

Canesi, Laura; Pruzzo, Carla; Tarsi, Renato; Gallo, Gabriella

2001-01-01

The role of type 1 fimbriae in the interactions between Escherichia coli and Mytilus galloprovincialis Lam. hemocytes was evaluated. The association of fimbriated strain MG155 with hemocyte monolayers at 18°C was 1.5- and 3- to 4-fold greater than the association of unfimbriated mutant AAEC072 in artificial seawater and in hemolymph serum, respectively. Such differences were apparently due to different adhesive properties since MG155 adhered more efficiently than AAEC072 when hemocytes were incubated at 4°C to inhibit the internalization process. Hemolymph serum increased both association and adherence of MG155 two- to threefold but did not affect association and adherence of AAEC072. MG155 was also 1.5- to 1.7-fold more sensitive to killing by hemocytes than AAEC072, as evaluated by the number of culturable bacteria after 60 and 120 min of incubation. The role of type 1 fimbriae in MG155 interactions with hemocytes was confirmed by the inhibitory effect of d-mannose. In in vivo experiments MG155 cells were cleared from circulating hemolymph more rapidly than AAEC072 cells were cleared. These results confirm that surface properties are crucial in influencing bacterial persistence and survival within mussel hemolymph. PMID:11133482

Effects of the popular food additive sodium benzoate on neural tube development in the chicken embryo.

PubMed

Emon, Selin Tural; Orakdogen, Metin; Uslu, Serap; Somay, Hakan

2015-01-01

Many more additives have been introduced with the development of processed foods. Neural tube defects are congenital malformations of the central nervous system. More than 300 000 children are born with neural tube defects every year and surviving children remain disabled for life. Sodium benzoate is used intensively in our daily lives. We therefore aimed to evaluate the effects of sodium benzoate on neural tube defects in chicken embryos. Fertile, specific pathogen-free eggs were used. The study was conducted on five groups. After 30 hours of incubation, the eggs were opened under 4x optical magnification. The embryonic disc was identified and sodium benzoate solution was injected. Eggs were closed with sterile adhesive strips and incubation was continued till the end of the 72nd hour. All eggs were then reopened and embryos were dissected from embryonic membranes and evaluated histopathologically. We found that the development of all embryos was consistent with the stage. We detected neural tube obstruction in one embryo. Neural tube defects were not detected in any embryos. This study showed that sodium benzoate as one of the widely used food preservatives has no effect to neural tube defect development in chicken embryos even at high doses.

Stable Isotopic Tracking of Autocthonous Carbon in Two Contrasting Ozark Streams

NASA Astrophysics Data System (ADS)

Ziegler, S.; Brisco-Townsend, S.

2005-05-01

The central role of microbes in biogeochemical processes makes the identification of carbon (C) sources fueling microorganisms critical to our understanding of stream ecosystems. The δ13C of biofilm phospholipid fatty acids (δ13CBPLFA) were determined in experiments conducted from July 2002 through July 2003 using 13C-labeled bicarbonate to track autochthonous C in two streams. In Moore Creek (MC), an agricultural stream, and Huey Hollow (HH), a forested stream, dissolved organic carbon (DOC) was released in light incubations during all seasons and represented >10% biofilm net primary production. The DOC from light incubations was enriched in 13C relative to DOC from dark incubations suggesting algal exudates were a major source of the DOC. The δ13CBPLFA suggest that 13C enriched exudates were not utilized by heterotrophic bacterial components in MC. Autotrophic PLFA from light incubations were more enriched in 13C while heterotrophic bacterial δ13CBPLFA were similar between light and dark incubations. By contrast, both heterotrophic and autotrophic biomarkers were significantly enriched in 13C in light incubations relative to dark incubations conducted in spring and summer in HH. Results suggest the exchange of C between autotrophic and heterotrophic components of biofilm communities differs between nutrient-enriched and depleted streams.

Effect of sex sorting on stallion spermatozoa: Heterologous oocyte binding, tyrosine phosphorylation and acrosome reaction assay.

PubMed

Balao da Silva, C M; Spinaci, M; Bucci, D; Giaretta, E; Peña, F J; Mari, G; Galeati, G

2013-09-01

The interest on sex sorting by flow cytometry on the equine industry has been increasing over the years. In this work, three different tests were performed in order to evaluate the membrane status of sorted stallion spermatozoa: assessment of binding ability to porcine oocytes, evaluation of acrosome integrity after stimulation with A23187, and detection of tyrosine phosphorylation. These evaluations were made after incubation for 0h, 1.5h and 3h in a capacitating medium. Sorted stallion spermatozoa attached similarly to the porcine oocytes, when compared with control samples. Sorted spermatozoa were more prone to undergo acrosome reaction (P<0.05), at the beginning and after 1.5h and 3h of incubation, and also had higher tyrosine phosphorylation of the tail (P<0.001), only at the beginning of the incubation period. Apparently sex sorted stallion spermatozoa are in a more advanced status of membrane destabilization, which could be associated with capacitation, although similar binding ability to porcine oocytes is maintained. Copyright © 2013 Elsevier B.V. All rights reserved.

A Liquid-Handling Robot for Automated Attachment of Biomolecules to Microbeads.

PubMed

Enten, Aaron; Yang, Yujia; Ye, Zihan; Chu, Ryan; Van, Tam; Rothschild, Ben; Gonzalez, Francisco; Sulchek, Todd

2016-08-01

Diagnostics, drug delivery, and other biomedical industries rely on cross-linking ligands to microbead surfaces. Microbead functionalization requires multiple steps of liquid exchange, incubation, and mixing, which are laborious and time intensive. Although automated systems exist, they are expensive and cumbersome, limiting their routine use in biomedical laboratories. We present a small, bench-top robotic system that automates microparticle functionalization and streamlines sample preparation. The robot uses a programmable microcontroller to regulate liquid exchange, incubation, and mixing functions. Filters with a pore diameter smaller than the minimum bead diameter are used to prevent bead loss during liquid exchange. The robot uses three liquid reagents and processes up to 10(7) microbeads per batch. The effectiveness of microbead functionalization was compared with a manual covalent coupling process and evaluated via flow cytometry and fluorescent imaging. The mean percentages of successfully functionalized beads were 91% and 92% for the robot and manual methods, respectively, with less than 5% bead loss. Although the two methods share similar qualities, the automated approach required approximately 10 min of active labor, compared with 3 h for the manual approach. These results suggest that a low-cost, automated microbead functionalization system can streamline sample preparation with minimal operator intervention. © 2015 Society for Laboratory Automation and Screening.

Mercury conversion processes in Amazon soils evaluated by thermodesorption analysis.

PubMed

do Valle, Cláudia M; Santana, Genilson P; Windmöller, Cláudia C

2006-12-01

This paper reports on the speciation study and the Hg redox behavior in Amazon soils not influenced by gold mining and collected near Manaus, AM, Brazil. The samples were incubated by adding Hg(0) and HgCl(2) to dry soil. Solid phase Hg speciation analysis was carried out using a Hg thermodesorption technique with the aim of distinguishing elemental Hg(0) from Hg(II) binding forms. In the first case, we observed the conversion of Hg(0) to Hg(II) binding forms in the range of 28-68% and a correlation between the percent of oxidation and OM content. Samples incubated with Hg(II) showed the formation of Hg(I) and/or Hg(0) in the range of 19-69%. The lowest values corresponded to the samples with the lowest clay contents. The kinetics of conversion of Hg(0) as well as HgCl(2) were roughly fitted to the two first order reactions, a fast one and a slow one. It was not possible to evaluate differences between sampling sites and types of soils, but the mean half-life of the first order reaction obtained by the addition of Hg(II) was slower (t(1/2)=365d) than the one obtained by the addition of Hg(0) (t(1/2)=148d). Previous studies have shown the predominance of organically bound Hg in these samples. Thus, the kinetic difference between Hg oxidation and reduction in combination with the efficient retention processes by OM may explain the high background values found in Amazon soils.

The effects of pre-incubation period and norharman on the mutagenic potency of 4-dimethylaminoazobenzene and 3'-methyl-4-dimethylaminoazobenzene in S. typhimurium.

PubMed

Lefevre, P A; Ashby, J

1981-01-01

The effects of the co-mutagen norharman on the mutagenicity of the rodent liver carcinogens 4-dimethyl-aminoazobenzene (DAB) and 3'-methyl-4-dimethyl-aminoazobenzene (3'-MeDAB) have been evaluated using the Ames Salmonella mutation assay. A period of pre-incubation was found to be necessary in order to detect DAB as a mutagen and to increase the initially weak response observed for 3'MeDAB; maximum responses were seen after 1 h pre-incubation in each case. The co-mutagenic properties of norharman were at their maximum for both azo-dyes in the direct plate-incorporation assay. Pre-incubation of either compound with norharman for increasing periods of time resulted in a decrease in potentiation, until after 1 h, an inhibition of mutagenicity was observed.

Benzoic Acid Production with Respect to Starter Culture and Incubation Temperature during Yogurt Fermentation using Response Surface Methodology.

PubMed

Yu, Hyung-Seok; Lee, Na-Kyoung; Jeon, Hye-Lin; Eom, Su Jin; Yoo, Mi-Young; Lim, Sang-Dong; Paik, Hyun-Dong

2016-01-01

Benzoic acid is occasionally used as a raw material supplement in food products and is sometimes generated during the fermentation process. In this study, the production of naturally occurring yogurt preservatives was investigated for various starter cultures and incubation temperatures, and considered food regulations. Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus plantarum, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium infantis, and Bifidobacterium breve were used as yogurt starter cultures in commercial starters. Among these strains, L. rhamnosus and L. paracasei showed the highest production of benzoic acid. Therefore, the use of L. rhamnosus, L. paracasei, S. thermophilus, and different incubation temperatures were examined to optimize benzoic acid production. Response surface methodology (RSM) based on a central composite design was performed for various incubation temperatures (35-44℃) and starter culture inoculum ratios (0-0.04%) in a commercial range of dairy fermentation processes. The optimum conditions were 0.04% L. rhamnosus, 0.01% L. paracasei, 0.02% S. thermophilus, and 38.12℃, and the predicted and estimated concentrations of benzoic acid were 13.31 and 13.94 mg/kg, respectively. These conditions maximized naturally occurring benzoic acid production during the yogurt fermentation process, and the observed production levels satisfied regulatory guidelines for benzoic acid in dairy products.

Benzoic Acid Production with Respect to Starter Culture and Incubation Temperature during Yogurt Fermentation using Response Surface Methodology

PubMed Central

Yoo, Mi-Young; Lim, Sang-Dong

2016-01-01

Benzoic acid is occasionally used as a raw material supplement in food products and is sometimes generated during the fermentation process. In this study, the production of naturally occurring yogurt preservatives was investigated for various starter cultures and incubation temperatures, and considered food regulations. Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus plantarum, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium infantis, and Bifidobacterium breve were used as yogurt starter cultures in commercial starters. Among these strains, L. rhamnosus and L. paracasei showed the highest production of benzoic acid. Therefore, the use of L. rhamnosus, L. paracasei, S. thermophilus, and different incubation temperatures were examined to optimize benzoic acid production. Response surface methodology (RSM) based on a central composite design was performed for various incubation temperatures (35-44℃) and starter culture inoculum ratios (0-0.04%) in a commercial range of dairy fermentation processes. The optimum conditions were 0.04% L. rhamnosus, 0.01% L. paracasei, 0.02% S. thermophilus, and 38.12℃, and the predicted and estimated concentrations of benzoic acid were 13.31 and 13.94 mg/kg, respectively. These conditions maximized naturally occurring benzoic acid production during the yogurt fermentation process, and the observed production levels satisfied regulatory guidelines for benzoic acid in dairy products. PMID:27433115

Evaluation of the Relationship between the Incubation Time and Carotenoid Production in Rhodotorula Slooffiae and R. Mucilaginosa Isolated from Leather Tanning Wastewater .

PubMed

Sadat Naghavi, Farzaneh; Hanachi, Parichehr; Soudi, Mohammad Reza; Saboora, Azra; Ghorbani, Atefeh

2013-10-01

Carotenoids which are naturally synthesized by fungi such as yeasts can act as an antioxidant which is closely related to their ability to decrease the risk of a variety of degenerative diseases. In recent years, the increase of demand for carotenoids obtained from natural sources has promoted major efforts to improve carotenoid production from biological sources such as pigmented yeasts. The aim of this study was comparing incubation time and carotenoid production in Rhodotorula slooffiae and R. mucilaginosa isolated from leather tanning wastewater. To isolate the carotenoid pigment, cells were suspended in acetone and broken using a homogenizer, followed by centrifugation and separation of supernatant. In order to study the effect of incubation time, samples were held at 30 ˚С in a shaker at 150 rpm for 24, 48, 72, 96, and 120 hr. For analytical evaluation, pigments were measured spectrophotometrically at 450 nm using the extinction coefficient E(1%) 450=2500. The results showed that the content of total carotenoid in R. slooffiae was the highest when samples were incubated for 72 hr. Overall, R. mucilaginosa had more potential to produce carotenoid. The best incubation periods for R. slooffiae and R. mucilaginosa were 72 hr and 48 hr, respectively. It seemed that the maximum rate of total carotenoid was not directly associated with the maximum amount of cell biomass and the type of carotenoid and their relative amount may vary depending on genus of yeast.

Drug Release as a function of bioactivity, incubation regime, liquid, and initial load: Release of bortezomib from calcium phosphate-containing silica/collagen xerogels.

PubMed

Kruppke, Benjamin; Hose, Dirk; Schnettler, Reinhard; Seckinger, Anja; Rößler, Sina; Hanke, Thomas; Heinemann, Sascha

2018-04-01

The ability of silica-/collagen-based composite xerogels to act as drug delivery systems was evaluated by taking into account the initial drug concentration, bioactivity of the xerogels, liquid, and incubation regime. The proteasome inhibitor bortezomib was chosen as a model drug, used for the systemic treatment of multiple myeloma. Incubation during 14 days in phosphate-buffered saline (PBS) or simulated body fluid (SBF) showed a weak initial burst and was identified to be of first order with subsequent release being independent from the initial load of 0.1 or 0.2 mg bortezomib per 60 mg monolithic sample. Faster drug release occurred during incubation in SBF compared to PBS, and during static incubation without changing the liquid, compared to dynamic incubation with daily liquid changes. Drug-loaded xerogels with hydroxyapatite as a third component exhibited enhanced bioactivity retarding drug release, explained by formation of a surface calcium phosphate layer. The fastest release of 50% of the total drug load was observed for biphasic xerogels after 7 days during dynamic incubation in SBF. As a result, the presented concept is suitable for the intended combination of the advantageous bone substitution properties of xerogels and local application of drugs exemplified by bortezomib. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1165-1173, 2018. © 2017 Wiley Periodicals, Inc.

Composition and location of simulated lake-shore redds influence incubation success in kokanee, Oncorhynchus nerka

USGS Publications Warehouse

Fincel, M.J.; Chipps, S.R.; Bennett, D.H.

2009-01-01

Methods for improving spawning habitat for lakeshore spawning kokanee, Oncorhynchus nerka (Walbaum), were explored by quantifying incubation success of embryos exposed to three substrate treatments in Lake Pend Oreille, Idaho, USA. Substrate treatments included no modification that used existing gravels in the lake (EXISTING), a cleaned substrate treatment where existing gravels were sifted in the water column to remove silt (CLEANED) and the addition of new, silt-free gravel (ADDED). Incubation success was evaluated using Whitlock-Vibert incubation boxes buried within each substrate treatment that contained recently fertilised embryos. Upon retrieval, live and dead sac fry and eyed eggs were enumerated to determine incubation success (sac fry and eyed eggs ?? 100/number of fertilised embryos). Incubation success varied significantly among locations and redd treatments. In general, incubation success among ADDED redds (0.0-13.0%) was significantly lower than that for EXISTING (1.4-61.0%) and CLEANED (0.4-62.5%) redds. Adding new gravel to spawning areas changed the morphometry of the gravel-water interface and probably exposed embryos to disturbance from wave action and reduced embryo survival. Moreover, efforts to improve spawning habitat for lakeshore spawning kokanee should consider water depth and location (e.g. protected shorelines) as important variables. Adding clean gravel to existing spawning areas may provide little benefit if water depth or lake-bottom morphometry are altered. ?? 2009 Blackwell Publishing Ltd.

The effects of heterospecifics and climatic conditions on incubation behavior within a mixed-species colony

USGS Publications Warehouse

Coates, Peter S.; Brussee, Brianne E.; Hothem, Roger L.; Howe, Kristy H.; Casazza, Michael L.; Eadie, John M.

2016-01-01

Parental incubation behavior largely influences nest survival, a critical demographic process in avian population dynamics, and behaviors vary across species with different life history breeding strategies. Although research has identified nest survival advantages of mixing colonies, behavioral mechanisms that might explain these effects is largely lacking. We examined parental incubation behavior using video-monitoring techniques on Alcatraz Island, California, of black-crowned night-heron Nycticorax nycticorax(hereinafter, night-heron) in a mixed-species colony with California gulls Larus californicus and western gulls L. occidentalis. We first quantified general nesting behaviors (i.e. incubation constancy, and nest attendance), and a suite of specific nesting behaviors (i.e. inactivity, vigilance, preening, and nest maintenance) with respect to six different daily time periods. We employed linear mixed effects models to investigate environmental and temporal factors as sources of variation in incubation constancy and nest attendance using 211 nest days across three nesting seasons (2010–2012). We found incubation constancy (percent of time on the eggs) and nest attendance (percent of time at the nest) were lower for nests that were located < 3 m from one or more gull nest, which indirectly supports the predator protection hypothesis, whereby heterospecifics provide protection allowing more time for foraging and other self-maintenance activities. To our knowledge, this is the first empirical evidence of the influence of one nesting species on the incubation behavior of another. We also identified distinct differences between incubation constancy and nest attentiveness, indicating that these biparental incubating species do not share similar energetic constraints as those that are observed for uniparental species. Additionally, we found that variation in incubation behavior was a function of temperature and precipitation, where the strength of these effects was dependent on the time of day. Overall, these findings strengthen our understanding of incubation behavior and nest ecology of a colonial-nesting species.

Novel methods to assess the influence of diet and incubation time on the emissions of odors from beef and swine manures

USDA-ARS?s Scientific Manuscript database

The influence of diet type and time of incubation (fresh vs. overnight) on the emissions of odors from the feces of beef cattle and swine was evaluated by gas chromatography-mass spectral (GC-MS) analysis. Swine (n=12) and beef cattle (n=24) were fed with two and four separate diets, respectively (n...

Developmental ossification sequences of the appendicular and axial skeleton in Kuttanad duck embryos (Anas platyrhynchos domesticus)

PubMed Central

Firdous, A.D.; Maya, S.; Massarat, K.; Baba, M.A.

2016-01-01

The processes of ossification sequences are poorly investigated for birds in general, even for domestic and experimental species and when it comes to the waterfowl it is almost negligible. Such sequences constitute a rich source of data on character evolution, and may even provide phylogenetic information. A pre-hatch developmental study on ossification sequences of axial and appendicular skeletal system in Kuttanad duck embryos was undertaken using 78 viable embryos. From day 3 to day 7 of incubation no ossification densities were seen both by alizarin red staining and computerized radiography. The first indication of ossification as small ossification centers in skull bones, clavicle, scapula, humerus, radius and ulna in forelimb and ilium, pubis femur and fibula in hind limb were observed on the 9th day of incubation. The ossification of the body of the ribs started at the 11th day of incubation towards the proximal extremity. On day 13th the ossification process of vertebrae was started from cervical end. The variation in appearance of the ossification centers in different bones at different stages of incubation period suggests relative importance of phylogeny to the sequences. PMID:26862514

No evidence of temperature-dependent sex determination or sex-biased embryo mortality in the chicken.

PubMed

Collins, K E; Jordan, B J; McLendon, B L; Navara, K J; Beckstead, R B; Wilson, J L

2013-12-01

Skewing the sex ratio at hatch in commercial poultry would be economically beneficial to the poultry industry. The existence of temperature-dependent sex determination is uncertain in birds. This experiment investigated if incubation temperatures skew sex ratios of commercial broilers. Three incubators were each set at a hot (38.3°C), standard (37.5°C), or cool (36.7°C) single-stage incubation temperature one time over 3 trials to eliminate incubator effect as a Latin square design. Sex ratios of hatched chicks and dead embryos were monitored. In one trial, embryo weights were evaluated. The percentages of male hatched chicks did not differ based on incubation temperature (P = 0.4486; 49.5% in the hot treatment, 51.4% at standard temperature, and 49.8% in the cool treatment). The percent hatch of eggs set was lower in the hot treatment (83.6%) than the standard (93.5%) and cool (91.6%) treatments (P < 0.0001) with greater late embryonic mortality in the hot treatment (P < 0.0001); however, the sex ratio of dead embryos did not differ among treatments (P = 0.9863). Pooled data of embryo mortality found no sex-biased embryo mortality with a female/male sex ratio of 1.22:1 (χ(2) = 1.27; P = 0.2596). Embryos from the hot treatment were heavier than those from the standard treatment by d 14 of incubation and were heavier than the embryos from the cool treatment by d 9 of incubation (P < 0.0001). These data indicate that incubation temperature affects embryonic mortality and embryonic growth rate, but it does not affect the sex ratio of broiler chickens. Additionally, no evidence was found for sex-biased embryo mortality in commercial broilers even at the incubation temperatures of this study.

Particle size alterations of feedstuffs during in situ neutral detergent fiber incubation.

PubMed

Krämer, M; Nørgaard, P; Lund, P; Weisbjerg, M R

2013-07-01

Particle size alterations during neutral detergent fiber (NDF) determination and in situ rumen incubation were analyzed by dry sieving and image analysis to evaluate the in situ procedure for estimation of NDF degradation parameters and indigestible NDF concentration in terms of particle size. Early-cut and late-cut grass silages, corn silage, alfalfa silage, rapeseed meal, and dried distillers grains were examined. Treatments were (1) drying and grinding of forage samples and grinding of concentrates; (2) neutral detergent-soluble (NDS) extraction; (3) machine washing and NDS extraction; (4) 24-h rumen incubation, machine washing, and NDS extraction; and (5) 288-h rumen incubation, machine washing, and NDS extraction. Degradation profiles for potentially degradable NDF were determined and image analysis was used to estimate particle size profiles and thereby the risk for particle loss. Particle dimensions changed during NDF determination and in situ rumen incubation and variations depended on feedstuff and treatment. Corn silage and late-cut grass silage varied most in particle area among feedstuffs, with an increase of 139% between 0 and 24h and a decrease of 77% between 24 and 288 h for corn silage and a decrease of 74% for late-cut grass silage between 24- and 288-h in situ rumen incubation. Especially for late-cut grass silage residues after 288 h in situ rumen incubation, a high mass proportion in the critical zone for escape was found. Particle area decreased linearly with increasing incubation time. Particle loss during in situ rumen incubation cannot be excluded and is likely to vary among feedstuffs. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Environmental Factors Affecting Ammonium Oxidation Under Iron Reducing Conditions

NASA Astrophysics Data System (ADS)

Jaffe, P. R.; Huang, S.; Ruiz-Urigüen, M.

2014-12-01

Ammonium (NH4+) oxidation coupled to iron (Fe) reduction in the absence of oxygen and nitrate/nitrite (NO3-/NO2-) has been reported by several investigators and referred to as Feammox. Feammox is a biological reaction, where Fe(III) is the electron acceptor, which is reduced to Fe(II), and NH4+ is the electron donor, which is oxidized to NO2-. Through a 180-day anaerobic incubation experiment, and using PCR-DGGE, 454-pyosequecing and qPCR analysis, we have shown that an Acidimicrobiaceae bacterium A6, a previously unreported species in the Acidimicrobiaceae family, might be either responsible or plays a key role in the Feammox process, We have enriched these Feammox bacteria (65.8% in terms of cell numbers) in a membrane reactor, and isolated the pure Acidimicrobiaceae bacterium A6 strain in an autotrophic medium. In samples collected and then incubated from a series of local wetland-, upland-, as well as storm-water detention pond-sediments, Feammox activity was only detected in acidic soil environments that contain Fe oxides. Using primers we developed for this purpose, Acidimicrobiaceae bacterium A6 was detected in all incubations where Feammox was observed. Anaerobic incubations of Feammox enrichment cultures adjusted to different pH, revealed that the optimal pH for Feammox is 4 ~ 5, and the reaction does not proceed when pH > 7. Feammox was still proceeding at pH as low as 2. In Feammox culture amended with different Fe(III) sources, Feammox reaction proceeded only when Fe oxides (ferrihydrite or goethite ) were supplied, whereas samples incubated with ferric chloride or ferric citrate showed no measurable NH4+ oxidation. Furthermore, we have also determined from incubation experiments conducted with a temperature gradient (10 ~ 35℃), that the Feammox process was active when the temperature is above 15℃, and the optimal temperature is 20℃. Incubations of enrichment culture with 79% Feammox bacteria appeared to remove circa 8% more NH4+ at 20ºC than at 35ºC. This is in contrast to anammox, another anaerobic ammonium oxidation pathway, for which optimal NH4+ oxidation is at temperatures ~ 30ºC. Hence, a Feammox-based process is an attractive candidate for wastewater treatment that could result in further energy savings, by requiring no aeration or heating of the wastewater in temperate climates.

Patterns relationships of student’s creativity with its indicators in learning optical instrument

NASA Astrophysics Data System (ADS)

Sukarmin; Dhian, T. E. V.; Nonoh, S. A.; Delisma, W. A.

2017-01-01

This study aims to identify patterns relationships of student’s creativity with its indicators in Learning Optical Instrument. The study was conducted at SMPN 2 Sawo. SMPN 1 Jetis, SMPIT Darut Taqwa, SMPN 1 Dander, Bojonegoro and SMPN 3 Plus Al-Fatima. Data analysis used descriptive analysis using the Confirmatory Factor Analysis. Creativity test instruments used have been tested parameters. Creativity indicators used are personal (self-confidence, perseverance), press (spirit, unyielding), process (preparation, incubation illumination, verification) and the product (knowledge, skills). Research Result shows that perseverance and incubation are the highest capabilities and verification capabilities of the lowest. All indicators on student creativity can still be improved. The relationship between creativity with the indicators grouped into a strong, moderate, weak and no relation. Indicators that have a strong relationship (r ≥ 0.50), namely are personal (self-confidence, perseverance), process (illumination). Indicators that have a connection was (0.3 ≤ r ≤ 0.49) are press (spirit), process (verification). Indicators which have a very low correlation (r ≤ 0.1 ≤ 0.29) are press (unyielding), process (preparation), process (incubation), product (skills) as shown in Figure 1. Indicators that do not have a relationship between the creativity of the students with the indicator that is, product (knowledge).

Evaluating AIDS Prevention: Contributions of Multiple Disciplines.

ERIC Educational Resources Information Center

Leviton, Laura C., Ed.; And Others

1990-01-01

Seven essays on efforts of evaluate prevention programs aimed at the acquired immune deficiency syndrome (AIDS) are presented. Topics include public health psychology, mathematical models of epidemiology, estimates of incubation periods, ethnographic evaluations of AIDS prevention programs, an AIDS education model, theory-based evaluation, and…

Nanoscale plasma chemistry enables fast, size-selective nanotube nucleation.

PubMed

Ostrikov, Kostya Ken; Mehdipour, Hamid

2012-03-07

The possibility of fast, narrow-size/chirality nucleation of thin single-walled carbon nanotubes (SWCNTs) at low, device-tolerant process temperatures in a plasma-enhanced chemical vapor deposition (CVD) is demonstrated using multiphase, multiscale numerical experiments. These effects are due to the unique nanoscale reactive plasma chemistry (NRPC) on the surfaces and within Au catalyst nanoparticles. The computed three-dimensional process parameter maps link the nanotube incubation times and the relative differences between the incubation times of SWCNTs of different sizes/chiralities to the main plasma- and precursor gas-specific parameters and explain recent experimental observations. It is shown that the unique NRPC leads not only to much faster nucleation of thin nanotubes at much lower process temperatures, but also to better selectivity between the incubation times of SWCNTs with different sizes and chiralities, compared to thermal CVD. These results are used to propose a time-programmed kinetic approach based on fast-responding plasmas which control the size-selective, narrow-chirality nucleation and growth of thin SWCNTs. This approach is generic and can be used for other nanostructure and materials systems. © 2012 American Chemical Society

STS-43 MS Lucid works with BIMDA-02 cell syringes on OV-104's middeck

NASA Image and Video Library

1991-08-11

STS043-03-001 (2-11 Aug 1991) --- Astronaut Shannon W. Lucid, STS-43 mission specialist, is pictured with a sample from the Bio-serve Instrumentation Technology Associates Materials Dispersion Apparatus (BIMDA). BIMDA is designed to obtain data on scientific methods and commercial potential for growing large high quality protein crystals in microgravity. The experimental focus is on both synthetic and natural biological processes that provide the foundation of the assembly of large structures from macromolecules. In addition, cell processes and membrane (cell and artificial) processes are being evaluated. BIMDA experiments are stored and operated on the middeck in a refrigerator/incubator module (R/IM). During this flight, the R/IM maintains a constant internal temperature of 20 degrees Celsius. This experiment also flew on NASA?s STS-37 mission.

Fluidized bed combustion residue as an alternative liming material and Ca source. [Prunus persica

DOE Office of Scientific and Technical Information (OSTI.GOV)

Edwards, J.H.; Horton, B.D.; White, A.W. Jr.

1985-01-01

Fluidized bed combustion residue (FBCR), a by-product of fossil fuel fired boilers, was evaluated as a liming material and a source of calcium for peaches (Prunus persica (L.) Batsch). Incubation studies involving a medium textured soil indicated that FBCR (calcite (FBCRC) or dolomitic (FBCRD) sources) was as effective a liming amendment as the respective agricultural limestone. Maximum soil pH occurred after 26 days incubation with FBCRC, but soil pH increased continuously throughout 137 days incubation with dolomitic limestone. Ammonium acetate extractable Ca was not affected by calcitic source, but Mg concentration increased with rates with the two dolomitic sources, andmore » was highest in the FBCRD source after 137 days incubation. In greenhouse studies with Elberta peach seedlings, FBCRC was more effective in neutralizing soil acidity and increasing extractable soil Ca than calcitic limestone.« less

The protective effects of N-Acetl-cysteine, oxo-thiazolidine-carboxylate, acetaminophen and their combinations against sulfur mustard cytotoxicity on human skin fibroblast cell line (HF2FF).

PubMed

Saberi, Mehdi; Zaree Mahmodabady, Ali

2009-10-01

Using human skin-fibroblast cell line HF2FF, the efficacy of some drugs was evaluated against sulfur mustard (SM) cytotoxicity. The drugs were the sulfhydryl containing molecule including N-acetylcysteine (NAC), 2-oxo-thiazolidine-4-carboxylate (OTC) and acetaminophen as glutathione (GSH) stimulator pathway. The protective effects of NAC (0.1 mM), OTC (1.8 mM), and acetaminophen (25 mM) alone or in combination with each other were evaluated on SM (180 M)-induced cytotoxicity. NAC and OTC were applied with SM simultaneously and acetaminophen 30 min before SM exposure, incubated for 1 h and then were rinsed and incubated with fresh medium. The efficacy was evaluated by determination of cells viability, intracellular GSH level and catalase activity 1 and 24 h post SM exposure or co-treatments. The cells viability was decreased 21.8% and 55.2%, respectively for 1 and 24 h post SM (1 h exposure) incubation. So, the 1-h SM exposure and 24-h treatment incubation were selected for evaluation. While, NAC alone treatment increased the cells viability (25%), GSH level (320%) and catalase activity (18%), the most effective combination was NAC plus OTC and acetaminophen which increased more significantly the cells viability (about 40%), GSH level (470%) and catalase activity (100%). The most effective combination was NAC (0.1 mM) plus OTC (1.8 mM) and acetaminophen (25 mM) which should be used before or concomitant with SM exposure. These drugs may reduce SM toxicity possibly by increment of GSH level and catalase activity. This efficacy needs to be confirmed by in vivo study.

In-ovo green light photostimulation during different embryonic stages affect somatotropic axis.

PubMed

Dishon, L; Avital-Cohen, N; Zaguri, S; Bartman, J; Heiblum, R; Druyan, S; Porter, T E; Gumulka, M; Rozenboim, I

2018-06-01

Previous studies demonstrated that in-ovo photostimulation with monochromatic green light increased the somatotropic axis expression in broilers embryos. The objective of the current study was to detect the critical period for in-ovo GL photostimulation, in order to find the optimal targeted photostimulation period during the incubation process. Three hundred thirty-six fertile broiler eggs were divided into 4 groups. The first group was incubated under dark conditions as a negative control. The second incubated under intermittent monochromatic green light using light-emitting diode (LED) lamps with an intensity of 0.1 W\\m2 at shell level from d 0 of the incubation as a positive control. The third group incubated under intermittent monochromatic green light from d 10 of the incubation. The last group incubated under intermittent monochromatic green light from d 15 of the incubation. In-ovo green light photostimulation from embryonic d 0 (ED0) increased plasma growth hormone (GH), as well as hypothalamic growth hormone releasing hormone (GHRH) and liver growth hormone receptor (GHR) and insulin-like growth factor-1 (IGF-1) mRNA levels. In-ovo green light photostimulation from ED10 increased the GH plasma levels compared to the negative control group, without affecting somatotropic axis mRNA genes expressions of GHRH, GHR, and IGF-1. In-ovo green light photostimulation from ED15 caused an increase in both the plasma GH levels and the somatotropic axis mRNA genes expressions of GHRH, GHR, and IGF-1, compared to the negative control group. These results suggest that the critical period of somatotropic axis acceleration by GL photostimulation start at 15 d of incubation.

High-throughput, 384-well, LC-MS/MS CYP inhibition assay using automation, cassette-analysis technique, and streamlined data analysis.

PubMed

Halladay, Jason S; Delarosa, Erlie Marie; Tran, Daniel; Wang, Leslie; Wong, Susan; Khojasteh, S Cyrus

2011-08-01

Here we describe a high capacity and high-throughput, automated, 384-well CYP inhibition assay using well-known HLM-based MS probes. We provide consistently robust IC(50) values at the lead optimization stage of the drug discovery process. Our method uses the Agilent Technologies/Velocity11 BioCel 1200 system, timesaving techniques for sample analysis, and streamlined data processing steps. For each experiment, we generate IC(50) values for up to 344 compounds and positive controls for five major CYP isoforms (probe substrate): CYP1A2 (phenacetin), CYP2C9 ((S)-warfarin), CYP2C19 ((S)-mephenytoin), CYP2D6 (dextromethorphan), and CYP3A4/5 (testosterone and midazolam). Each compound is incubated separately at four concentrations with each CYP probe substrate under the optimized incubation condition. Each incubation is quenched with acetonitrile containing the deuterated internal standard of the respective metabolite for each probe substrate. To minimize the number of samples to be analyzed by LC-MS/MS and reduce the amount of valuable MS runtime, we utilize timesaving techniques of cassette analysis (pooling the incubation samples at the end of each CYP probe incubation into one) and column switching (reducing the amount of MS runtime). Here we also report on the comparison of IC(50) results for five major CYP isoforms using our method compared to values reported in the literature.

Noise levels in a neonatal transport incubator in medically configured aircraft.

PubMed

Sittig, Steven E; Nesbitt, Jeffrey C; Krageschmidt, Dale A; Sobczak, Steven C; Johnson, Robert V

2011-01-01

The purpose of this study was to evaluate exposure of neonates to noise during air medical transport as few commercially available hearing protective devices exist for premature newborns during air medical transport. Sound pressure levels in an infant incubator during actual flight conditions in four common medically configured aircraft were measured. Three noise dosimeters measured time-weighted average noise exposure during flight in each aircraft. One dosimeter was placed in the infant incubator, and the remaining dosimeters recorded noise levels in various parts of the aircraft cabin. The incubator provided a 6-dBA decrease in noise exposure from that in the crew cabin. The average noise level in the incubator in all aircraft was close to 80 dB, much higher than the proposed limits of 45 dB for neonatal intensive care unit noise exposure or 60 dB during transport. Exposure of neonates to elevated noise levels during transport may be harmful, and steps should be taken to protect the hearing of this patient population. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

Vitamin C acts as a hepatoprotectant in carbofuran treated rat liver slices in vitro.

PubMed

Jaiswal, Sunil Kumar; Gupta, Vivek Kumar; Ansari, Md Dilshad; Siddiqi, Nikhat J; Sharma, Bechan

2017-01-01

Carbamates, most commonly used pesticides in agricultural practices, have been reported to produce free radicals causing deleterious effects in animals. The present study was designed to assess the carbofuran induced oxidative stress in rat liver slices in vitro and also to evaluate protective role of vitamin C by incubating them in Krebs-Ringer HEPES Buffer (KRHB) containing incubation media (Williams medium E (WME) supplemented with glucose and antibiotics) with different concentrations of carbofuran. The results demonstrated that carbofuran caused significant increase in lipid peroxidation and inhibition in the activity of hepatic superoxide dismutase (SOD) in concentration dependent manner. The data with incubation medium reflected that carbofuran at lowest concentration caused an increase in SOD activity followed by its inhibition at higher concentration. Carbofuran treatment caused inhibition in the activity of catalase in liver slices and WME incubation medium. Pre-incubation of liver slices and the WME media with vitamin C restored the values of biochemical indices tested. The results indicated that carbofuran might induce oxidative stress in hepatocytes. The pretreatment with vitamin C may offer hepatoprotection from toxicity of pesticide at low concentration only.

Physical origin of the incubation time of self-induced GaN nanowires

DOE Office of Scientific and Technical Information (OSTI.GOV)

Consonni, V.; Trampert, A.; Geelhaar, L.

2011-07-18

The nucleation process of self-induced GaN nanowires grown by molecular beam epitaxy has been investigated by reflection high-energy electron diffraction measurements. It is found that stable nuclei in the form of spherical cap-shaped islands develop only after an incubation time that is strongly dependent upon the growth conditions. Its evolution with the growth temperature and gallium rate has been described within standard island nucleation theory, revealing a nucleation energy of 4.9 {+-} 0.1 eV and a very small nucleus critical size. The consideration of the incubation time is critical for the control of the nanowire morphology.

Interactions between seed traits and digestive processes determine the germinability of bird-dispersed seeds

PubMed Central

Soons, Merel B.

2018-01-01

Waterbirds disperse a wide range of plant seeds via their guts, promoting biotic connectivity between isolated habitat patches. However, the intensity of digestive forces encountered by seeds, and therefore their potential to survive digestive tract passage, varies within and between waterbird species. Here, we investigate under controlled conditions how the interaction between seed traits and digestive strategies affect the germinability of seeds following waterbird-mediated dispersal. We exposed seeds of 30 wetland plant species to the main digestive processes in the dabbling duck digestive system: mechanical, chemical and intestinal digestion. These were simulated by 1) a pressure test and scarification treatment, 2) incubation in simulated gastric juice, and 3) incubation in intestinal contents of culled mallards (Anas platyrhynchos). We evaluated their separate and combined effects on seed germination, and identified the role of seed size and seed coat traits in resisting the digestive forces. Seeds were generally resistant to separate digestive processes, but highly sensitive to a combination. Resistance to mechanical break-down was reduced by up to 80% by chemical pre-treatment, especially for seeds with permeable coats. Scarified seeds were 12–17% more vulnerable to chemical and intestinal digestive processes than undamaged seeds. Large seeds and seeds with thin, permeable coats were particularly sensitive to chemical and intestinal digestion. These results indicate that efficient digestion of seeds requires multiple digestive processes. The gizzard, responsible for mechanical digestion, plays a key role in seed survival. Omnivorous birds, which have relatively light gizzards compared to pure herbivores or granivores, are thus most likely to disperse seeds successfully. Regardless of digestive strategy, small seeds with tough seed coats are most resistant to digestion and may be adapted to endozoochorous dispersal by waterbirds. PMID:29614085

Interactions between seed traits and digestive processes determine the germinability of bird-dispersed seeds.

PubMed

Kleyheeg, Erik; Claessens, Mascha; Soons, Merel B

2018-01-01

Waterbirds disperse a wide range of plant seeds via their guts, promoting biotic connectivity between isolated habitat patches. However, the intensity of digestive forces encountered by seeds, and therefore their potential to survive digestive tract passage, varies within and between waterbird species. Here, we investigate under controlled conditions how the interaction between seed traits and digestive strategies affect the germinability of seeds following waterbird-mediated dispersal. We exposed seeds of 30 wetland plant species to the main digestive processes in the dabbling duck digestive system: mechanical, chemical and intestinal digestion. These were simulated by 1) a pressure test and scarification treatment, 2) incubation in simulated gastric juice, and 3) incubation in intestinal contents of culled mallards (Anas platyrhynchos). We evaluated their separate and combined effects on seed germination, and identified the role of seed size and seed coat traits in resisting the digestive forces. Seeds were generally resistant to separate digestive processes, but highly sensitive to a combination. Resistance to mechanical break-down was reduced by up to 80% by chemical pre-treatment, especially for seeds with permeable coats. Scarified seeds were 12-17% more vulnerable to chemical and intestinal digestive processes than undamaged seeds. Large seeds and seeds with thin, permeable coats were particularly sensitive to chemical and intestinal digestion. These results indicate that efficient digestion of seeds requires multiple digestive processes. The gizzard, responsible for mechanical digestion, plays a key role in seed survival. Omnivorous birds, which have relatively light gizzards compared to pure herbivores or granivores, are thus most likely to disperse seeds successfully. Regardless of digestive strategy, small seeds with tough seed coats are most resistant to digestion and may be adapted to endozoochorous dispersal by waterbirds.

Estimating the Delay between Host Infection and Disease (Incubation Period) and Assessing Its Significance to the Epidemiology of Plant Diseases

PubMed Central

Leclerc, Melen; Doré, Thierry; Gilligan, Christopher A.; Lucas, Philippe; Filipe, João A. N.

2014-01-01

Knowledge of the incubation period of infectious diseases (time between host infection and expression of disease symptoms) is crucial to our epidemiological understanding and the design of appropriate prevention and control policies. Plant diseases cause substantial damage to agricultural and arboricultural systems, but there is still very little information about how the incubation period varies within host populations. In this paper, we focus on the incubation period of soilborne plant pathogens, which are difficult to detect as they spread and infect the hosts underground and above-ground symptoms occur considerably later. We conducted experiments on Rhizoctonia solani in sugar beet, as an example patho-system, and used modelling approaches to estimate the incubation period distribution and demonstrate the impact of differing estimations on our epidemiological understanding of plant diseases. We present measurements of the incubation period obtained in field conditions, fit alternative probability models to the data, and show that the incubation period distribution changes with host age. By simulating spatially-explicit epidemiological models with different incubation-period distributions, we study the conditions for a significant time lag between epidemics of cryptic infection and the associated epidemics of symptomatic disease. We examine the sensitivity of this lag to differing distributional assumptions about the incubation period (i.e. exponential versus Gamma). We demonstrate that accurate information about the incubation period distribution of a pathosystem can be critical in assessing the true scale of pathogen invasion behind early disease symptoms in the field; likewise, it can be central to model-based prediction of epidemic risk and evaluation of disease management strategies. Our results highlight that reliance on observation of disease symptoms can cause significant delay in detection of soil-borne pathogen epidemics and mislead practitioners and epidemiologists about the timing, extent, and viability of disease control measures for limiting economic loss. PMID:24466153

Effects of maternal dietary manganese and incubation temperature on hatchability, antioxidant status, and expression of heat shock proteins in chick embryos.

PubMed

Zhu, Y W; Lu, L; Li, W X; Zhang, L Y; Ji, C; Lin, X; Liu, H C; Odle, J; Luo, X G

2015-12-01

To investigate whether supplementing manganese (Mn) to the maternal diet could reduce the deleterious effect of heat stress on the developing embryo, the hatchability, antioxidant status, and expression of heat shock proteins (HSP) were evaluated in chick embryos under normal and high incubation temperatures. A completely randomized design ( = 6) with 2 maternal dietary Mn treatments (unsupplemented control basal diet versus the basal diet + 120 mg Mn/kg as inorganic Mn) × 2 incubation temperatures (normal, 37.8°C, versus high, 39.0°C) was used. High incubation temperature did not affect ( > 0.19) hatchability and embryo mortality and development but did increase ( < 0.05) activities of heart manganese superoxide dismutase (MnSOD) and liver copper zinc superoxide dismutase and liver MnSOD mRNA and protein levels in embryos. High incubation temperature also decreased ( < 0.003) HSP70 protein level in the heart but had no effects ( > 0.07) in the liver of embryos. Maternal diet with Mn supplementation not only increased ( < 0.05) the hatchability and Mn content ( < 0.001) in the yolk and embryonic tissues and the activity of MnSOD in the heart ( < 0.004) as well as relative liver weight ( < 0.05) under normal incubation temperature but also decreased ( ≤ 0.05) embryo mortality and HSP90 mRNA level in the liver and heart of embryos. Furthermore, under high incubation temperature, maternal diet Mn supplementation increased ( < 0.002) MnSOD protein expression in the liver of embryos but had no effect ( > 0.43) under normal incubation temperature. These results indicated that high incubation temperature induced self-protective responses of chick embryos with a modification of antioxidant status and a depression of HSP70 protein level. Maternal dietary supplementation of Mn could improve the hatchability as well as antioxidant ability to protect against heat challenge in embryos during incubation.

Effects of ultrashort gamete co-incubation time on porcine in vitro fertilization.

PubMed

Almiñana, C; Gil, M A; Cuello, C; Parrilla, I; Roca, J; Vazquez, J M; Martinez, E A

2008-07-01

A reduction in co-incubation time has been suggested as an alternative method to reduce polyspermic fertilization. The aim of this study was to evaluate the effect of short periods of gamete co-incubation during pig in vitro fertilization. A total of 2,833 in vitro matured oocytes were inseminated with thawed spermatozoa and coincubated for 0.25, 1, 2, 3, 7, 10 min and 6h. The oocytes from the 0.25-10 min groups were washed three times in modified Tris-buffered medium (mTBM) medium to remove spermatozoa not bound to the zona and transferred to the same medium (containing no spermatozoa) until 6h of co-incubation time were completed. After 6h, presumptive zygotes from each group were cultured in NCSU-23 medium for 12-15 h to assess fertilization parameters. After each period of co-incubation, 45-50 oocytes from each group were stained with Hoechst-33342 and the number of spermatozoa bound to the zona was counted. Although the number of zona bound spermatozoa increased (p<0.05) with the co-incubation time, no increase was observed in penetration rates among groups from 2 min to 6h of co-incubation time (ranging from 53.5+/-2.8 to 61.3+/-2.6%). Similarly, the efficiency of fertilization reached a maximum for the 2 min of co-incubation group with values ranging between 32.3+/-2.4 and 41.9+/-2.5%. The reduction of co-incubation time did not affect the monospermy rate (range: 71.3+/-3.4-80.2+/-3.8%) and the mean number of spermatozoa/oocyte (range: 1.2+/-0.4-1.4+/-0.5). These results show that, under our in vitro conditions, high penetration rate can be obtained with co-incubation times as short as 2 min, although monospermy could not be improved using this strategy.

Estimating the delay between host infection and disease (incubation period) and assessing its significance to the epidemiology of plant diseases.

PubMed

Leclerc, Melen; Doré, Thierry; Gilligan, Christopher A; Lucas, Philippe; Filipe, João A N

2014-01-01

Knowledge of the incubation period of infectious diseases (time between host infection and expression of disease symptoms) is crucial to our epidemiological understanding and the design of appropriate prevention and control policies. Plant diseases cause substantial damage to agricultural and arboricultural systems, but there is still very little information about how the incubation period varies within host populations. In this paper, we focus on the incubation period of soilborne plant pathogens, which are difficult to detect as they spread and infect the hosts underground and above-ground symptoms occur considerably later. We conducted experiments on Rhizoctonia solani in sugar beet, as an example patho-system, and used modelling approaches to estimate the incubation period distribution and demonstrate the impact of differing estimations on our epidemiological understanding of plant diseases. We present measurements of the incubation period obtained in field conditions, fit alternative probability models to the data, and show that the incubation period distribution changes with host age. By simulating spatially-explicit epidemiological models with different incubation-period distributions, we study the conditions for a significant time lag between epidemics of cryptic infection and the associated epidemics of symptomatic disease. We examine the sensitivity of this lag to differing distributional assumptions about the incubation period (i.e. exponential versus Gamma). We demonstrate that accurate information about the incubation period distribution of a pathosystem can be critical in assessing the true scale of pathogen invasion behind early disease symptoms in the field; likewise, it can be central to model-based prediction of epidemic risk and evaluation of disease management strategies. Our results highlight that reliance on observation of disease symptoms can cause significant delay in detection of soil-borne pathogen epidemics and mislead practitioners and epidemiologists about the timing, extent, and viability of disease control measures for limiting economic loss.

Short-term incubation studies on degradation of biochar in soil

NASA Astrophysics Data System (ADS)

Lanza, Giacomo; Wirth, Stephan; Geßler, Arthur; Kern, Jürgen; Mumme, Jan

2014-05-01

Biochar is considered a stable, recalcitrant substance, which holds potential to store carbon in soils for prolonged time and therefore would provide a long-term carbon sink. Furthermore, biochar is discussed to enhance soil fertility and plant productivity, and may improve water and nutrient holding capacity. However, mineralisation to CO2 may occur, as for any soil organic carbon pool, depending on char composition, soil properties and environmental conditions. Therefore, it is important to gain insight into the stability of its carbon structure and the dynamics of decay processes in soil. The evaluation of biochar stability in soil is complicated by the impact of external factors thus as soil moisture and temperature, soil nutrient status and moreover by extended decay timescales. To overcome these difficulties, we performed dynamic incubation experiments under laboratory conditions, using a multi-channel, automated infra-red gas analysis system at 20°C for up to 10 days to detect CO2 emission over time. Our aim was to compare the decay dynamics of different biochar preparations added to soil, i.e. HTC-char and pyrochar from maize silage with and without biological post-processing (anaerobic digestion), as compared to unmodified maize straw. Digestate from a maize silage-fed anaerobic biogas reactor was also tested. As a result, the addition of charred or digested materials to soil resulted in much lower CO2 emission rates as compared to the unmodified maize straw, proving stability of biochar carbon compounds. Pyrochar showed to be the most stable of all substrates added, as the CO2 emission was hardly distinguishable from that of the control soil. Soil enriched with HTC-char emitted significantly more CO2 compared to soil enriched with pyrochar, but the post-processing was effective in reducing the emissions. Furthermore, HTC-char showed a two-step decay kinetics, which cannot apparently be explained with a simple double-pool model. In conclusion, the short-term incubation approach was effective to highlight differences in decomposition dynamics between the considered substrates in soil, and confirmed the effectiveness of the charring process to increase the stability of organic substrates in soil. More investigations are necessary to reveal the impact of readily available substrates and nutrients on degradation of biochar in soil, and to clarify the mechanisms responsible for the observed kinetics in order to derive a suitable process model.

Nanoparticles containing allotropes of carbon have genotoxic effects on glioblastoma multiforme cells

PubMed Central

Hinzmann, Mateusz; Jaworski, Sławomir; Kutwin, Marta; Jagiełło, Joanna; Koziński, Rafał; Wierzbicki, Mateusz; Grodzik, Marta; Lipińska, Ludwika; Sawosz, Ewa; Chwalibog, Andrè

2014-01-01

The carbon-based nanomaterial family consists of nanoparticles containing allotropes of carbon, which may have a number of interactions with biological systems. The objective of this study was to evaluate the toxicity of nanoparticles comprised of pristine graphene, reduced graphene oxide, graphene oxide, graphite, and ultradispersed detonation diamond in a U87 cell line. The scope of the work consisted of structural analysis of the nanoparticles using transmission electron microscopy, evaluation of cell morphology, and assessment of cell viability by Trypan blue assay and level of DNA fragmentation of U87 cells after 24 hours of incubation with 50 μg/mL carbon nanoparticles. DNA fragmentation was studied using single-cell gel electrophoresis. Incubation with nanoparticles containing the allotropes of carbon did not alter the morphology of the U87 cancer cells. However, incubation with pristine graphene and reduced graphene oxide led to a significant decrease in cell viability, whereas incubation with graphene oxide, graphite, and ultradispersed detonation diamond led to a smaller decrease in cell viability. The results of a comet assay demonstrated that pristine graphene, reduced graphene oxide, graphite, and ultradispersed detonation diamond caused DNA damage and were therefore genotoxic in U87 cells, whereas graphene oxide was not. PMID:24876774

Inhibitory Effect of Veterinary Antibiotics on Denitrification in Groundwater: A Microcosm Approach

PubMed Central

Kim, Kangjoo; Cho, Ju-Sik; Lee, Young Han; Joo, Young Kyoo; Lee, Sang Soo

2014-01-01

Veterinary antibiotics in groundwater may affect natural microbial denitrification process. A microcosm study was conducted to evaluate the influence of sulfamethazine and chlortetracycline at different concentrations (0, 0.01, 0.1, and 1.0 mg/L) on nitrate reduction in groundwater under denitrifying condition. Decrease in nitrate removal and nitrite production was observed with the antibiotics. Maximum inhibition of nitrate removal was observed after seven days of incubation with 0.01 mg/L sulfamethazine (17.0%) and 1.0 mg/L chlortetracycline (15.4%). The nitrite production was inhibited with 1.0 mg/L sulfamethazine to 82.0% and chlortetracycline to 31.1%. The initial/final nitrate concentrations indicated that 0.01 mg/L sulfamethazine and 1.0 mg/L chlortetracycline were most effective in inhibiting activity of denitrifying bacteria in groundwater. After 12 days of incubation, the sulfamethazine biodegradation was observed whereas chlortetracycline was persistent. Sulfamethazine and chlortetracycline in groundwater could inhibit the growth and capability of naturally occurring denitrifying bacteria, thereby threatening nitrate pollution in groundwater. PMID:24757442

Changes in brain magnetic resonance imaging patterns for preterm infants after introduction of a magnetic resonance-compatible incubator coil system: 5-year experience at a single institution.

PubMed

Cho, Hyun-Hae; Kim, In-One; Cheon, Jung-Eun; Choi, Young Hun; Lee, So Mi; Kim, Woo Sun

2016-09-01

To evaluate the changes in using patterns of brain magnetic resonance imaging (MRI) in preterm infants after introduction of a MR-compatible incubator coil system. Brain MRIs for preterm infants with the MR-compatible incubator coil from March 2010 to July 2014 (n=154, group A) were compared with MRIs prior to the introduction of the incubator coil, from March 2005 to February 2010 (n=65, group B). Clinical data, MRI findings, acquisition time, and incidence of adverse events during the study were retrospectively reviewed. For the qualitative analysis of the examinations, the presence of motion artefact, spatial resolution, and overall image quality were assessed. Signal uniformity of each sequence was evaluated for a quantitative comparison. Comparing with group B, Group A was significantly younger (36+3 vs. 38+3 weeks, p<0.001), had a significantly lower body weight (2006.6 and 2390.3g respectively; p<0.001) at the time of MRI, and had shorter time interval (54.3±2.6 vs. 70.5±4.4days, p=0.002) between birth and examination. Abnormal findings were noted more frequently in group A (n=100, 65%) than in B (n=24, 37%. p=0.001) with a significantly higher incidence of diffusion restriction (n=21, 13.6% vs. n=4, 6.2%, p=0.034). Mean image acquisition time was significantly shorter in group A (21.4±4.5 vs. 25.4±5.5min, p<0.001) with significant lower adverse events during MRI (n=26, 40 vs. n=6, 3.9%, p<0.001). Group A exhibited significantly less motion artefact, better spatial resolution, and better overall image quality with decreased signal variation than group B (all p<0.001). Application of the MR-compatible incubator for preterm brain MRI evaluation is safer and provides more timely evaluation of preterm infants with better image quality. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Description of Aspergillus flavus growth under the influence of different factors (water activity, incubation temperature, protein and fat concentration, pH, and cinnamon essential oil concentration) by kinetic, probability of growth, and time-to-detection models.

PubMed

Kosegarten, Carlos E; Ramírez-Corona, Nelly; Mani-López, Emma; Palou, Enrique; López-Malo, Aurelio

2017-01-02

A Box-Behnken design was used to determine the effect of protein concentration (0, 5, or 10g of casein/100g), fat (0, 3, or 6g of corn oil/100g), a w (0.900, 0.945, or 0.990), pH (3.5, 5.0, or 6.5), concentration of cinnamon essential oil (CEO, 0, 200, or 400μL/kg) and incubation temperature (15, 25, or 35°C) on the growth of Aspergillus flavus during 50days of incubation. Mold response under the evaluated conditions was modeled by the modified Gompertz equation, logistic regression, and time-to-detection model. The obtained polynomial regression models allow the significant coefficients (p<0.05) for linear, quadratic and interaction effects for the Gompertz equation's parameters to be identified, which adequately described (R 2 >0.967) the studied mold responses. After 50days of incubation, every tested model system was classified according to the observed response as 1 (growth) or 0 (no growth), then a binary logistic regression was utilized to model A. flavus growth interface, allowing to predict the probability of mold growth under selected combinations of tested factors. The time-to-detection model was utilized to estimate the time at which A. flavus visible growth begins. Water activity, temperature, and CEO concentration were the most important factors affecting fungal growth. It was observed that there is a range of possible combinations that may induce growth, such that incubation conditions and the amount of essential oil necessary for fungal growth inhibition strongly depend on protein and fat concentrations as well as on the pH of studied model systems. The probabilistic model and the time-to-detection models constitute another option to determine appropriate storage/processing conditions and accurately predict the probability and/or the time at which A. flavus growth occurs. Copyright © 2016 Elsevier B.V. All rights reserved.

Effects of mixing eggs of different initial incubation time on the hatching pattern, chick embryonic development and post-hatch performance.

PubMed

Zhong, Zhentao; Yu, Yue; Jin, Shufang; Pan, Jinming

2018-01-01

The hatch window that varies from 24 to 48 h is known to influence post-hatch performance of chicks. A narrow hatch window is needed for commercial poultry industry to acquire a high level of uniformity of chick quality. Hatching synchronization observed in avian species presents possibilities in altering hatch window in artificial incubation. Layer eggs which were laid on the same day by a single breeder flock and stored for no more than two days started incubation 12 h apart to obtain developmental distinction. The eggs of different initial incubation time were mixed as rows adjacent to rows on day 12 of incubation. During the hatching period (since day 18), hatching time of individual eggs and hatch window were obtained by video recordings. Embryonic development (day 18 and 20) and post-hatch performance up to day 7 were measured. The manipulation of mixing eggs of different initial incubation time shortened the hatch window of late incubated eggs in the manipulated group by delaying the onset of hatching process, and improved the hatchability. Compared to the control groups, chick embryos or chicks in the egg redistribution group showed no significant difference in embryonic development and post-hatch performance up to day 7. We have demonstrated that eggs that were incubated with advanced eggs performed a narrow spread of hatch with higher hatchability, normal embryonic development as well as unaffected chick quality. This specific manipulation is applicable in industrial poultry production to shorten hatch window and improve the uniformity of chick quality.

Intraspecific Variation in and Environment-Dependent Resource Allocation to Embryonic Development Time in Common Terns.

PubMed

Vedder, Oscar; Kürten, Nathalie; Bouwhuis, Sandra

Embryonic development time is thought to impact life histories through trade-offs against life-history traits later in life, yet the inference is based on interspecific comparative analyses only. It is largely unclear whether intraspecific variation in embryonic development time that is not caused by environmental differences occurs, which would be required to detect life-history trade-offs. Here we performed a classical common-garden experiment by incubating fresh eggs of free-living common terns (Sterna hirundo) in a controlled incubation environment at two different temperatures. Hatching success was high but was slightly lower at the lower temperature. While correcting for effects of year, incubation temperature, and laying order, we found significant variation in the incubation time embryos required until hatching and in their heart rate. Embryonic heart rate was significantly positively correlated within clutches, and a similar tendency was found for incubation time, suggesting that intrinsic differences in embryonic development rate between offspring of different parents exist. Incubation time and embryonic heart rate were strongly correlated: embryos with faster heart rates required shorter incubation time. However, after correction for heart rate, embryos still required more time for development at the lower incubation temperature. This suggests that processes other than development require a greater share of resources in a suboptimal environment and that relative resource allocation to development is, therefore, environment dependent. We conclude that there is opportunity to detect intraspecific life-history trade-offs with embryonic development time and that the resolution of trade-offs may differ between embryonic environments.

Exo-polygalacturonase production by Bacillus subtilis CM5 in solid state fermentation using cassava bagasse

PubMed Central

Swain, Manas R.; Kar, Shaktimay; Ray, Ramesh C.

2009-01-01

The purpose of this investigation was to study the effect of Bacillus subtilis CM5 in solid state fermentation using cassava bagasse for production of exo-polygalacturonase (exo-PG). Response surface methodology was used to evaluate the effect of four main variables, i.e. incubation period, initial medium pH, moisture holding capacity (MHC) and incubation temperature on enzyme production. A full factorial Central Composite Design was applied to study these main factors that affected exo-PG production. The experimental results showed that the optimum incubation period, pH, MHC and temperature were 6 days, 7.0, 70% and 50°C, respectively for optimum exo-PG production. PMID:24031409

Short incubation fractional CO2 laser-assisted photodynamic therapy vs. conventional photodynamic therapy in field-cancerized skin: 12-month follow-up results of a randomized intraindividual comparison study.

PubMed

Vrani, F; Sotiriou, E; Lazaridou, E; Vakirlis, E; Sideris, N; Kirmanidou, E; Apalla, Z; Lallas, A; Ioannides, D

2018-06-04

Topical methyl aminolevulinate photodynamic therapy (MAL-PDT) with 3 h incubation is recommended as a field directed treatment. Skin pretreatment with ablative CO 2 fractional laser (AFXL) prior to MAL-PDT enhances drug penetration and could minimize incubation time. To evaluate and compare the safety and the preventive effect in the development of new non-melanocytic skin cancers (NMSCs) of AFXL-assisted MAL-PDT with 1-h incubation with that of conventional MAL-PDT in patients with clinical and histological signs of field cancerization. Forty-two patients with two mirror cancerized areas of face or scalp were randomized to field treatment with 1-h incubation AFXL-assisted PDT or conventional PDT (CPDT). All patients underwent two treatment sessions 1 week apart. Irradiation was performed using a red light-emitting diode lamp at 37 J/cm 2 . Patients were followed up at 3, 6, 9 and 12 months for the evaluation of development of new NMSCs lesions. All patients completed the study. There was no statistically significant difference with respect to the total number of new actinic keratoses at any point of follow-up as well as to the mean time of occurrence of new lesions between treatment fields. Both treatment regimens were safe and well tolerated. Ablative CO 2 fractional laser pretreatment may be considered as an option for reducing photosensitizer occlusion time while providing the same preventative efficacy as CPDT in patients with field-cancerized skin. © 2018 European Academy of Dermatology and Venereology.

Evaluation of the Relationship between the Incubation Time and Carotenoid Production in Rhodotorula Slooffiae and R. Mucilaginosa Isolated from Leather Tanning Wastewater 

PubMed Central

Sadat Naghavi, Farzaneh; Hanachi, Parichehr; Soudi, Mohammad Reza; Saboora, Azra; Ghorbani, Atefeh

2013-01-01

Objective(s): Carotenoids which are naturally synthesized by fungi such as yeasts can act as an antioxidant which is closely related to their ability to decrease the risk of a variety of degenerative diseases. In recent years, the increase of demand for carotenoids obtained from natural sources has promoted major efforts to improve carotenoid production from biological sources such as pigmented yeasts. The aim of this study was comparing incubation time and carotenoid production in Rhodotorula slooffiae and R. mucilaginosa isolated from leather tanning wastewater. Materials and Methods: To isolate the carotenoid pigment, cells were suspended in acetone and broken using a homogenizer, followed by centrifugation and separation of supernatant. In order to study the effect of incubation time, samples were held at 30 ˚С in a shaker at 150 rpm for 24, 48, 72, 96, and 120 hr. For analytical evaluation, pigments were measured spectrophotometrically at 450 nm using the extinction coefficient E1%450=2500. Results: The results showed that the content of total carotenoid in R. slooffiae was the highest when samples were incubated for 72 hr. Overall, R. mucilaginosa had more potential to produce carotenoid. The best incubation periods for R. slooffiae and R. mucilaginosa were 72 hr and 48 hr, respectively. Conclusion: It seemed that the maximum rate of total carotenoid was not directly associated with the maximum amount of cell biomass and the type of carotenoid and their relative amount may vary depending on genus of yeast. PMID:24379970

A comparison of in situ methods for measuring net nitrogen mineralization rates of organic soil amendments.

PubMed

Hanselman, Travis A; Graetz, Donald A; Obreza, Thomas A

2004-01-01

In situ incubation methods may help provide site-specific estimates of N mineralization from land-applied wastes. However, there are concerns about the reliability of the data generated by the various methods due to containment artifacts. We amended a sandy soil with either poultry manure, biosolids, or yard-waste compost and incubated the mixtures using four in situ methods (buried bags, covered cylinders, standard resin traps, and "new" soil-resin traps) and a conventional laboratory technique in plastic bags. Each incubation device was destructively sampled at 45-d intervals for 180 d and net N mineralization was determined by measuring the amount of inorganic N that accumulated in the soil or soil plus resin traps. Containment effects were evaluated by comparing water content of the containerized soil to a field-reference soil column. In situ incubation methods provided reasonable estimates of short-term (< 45 d) N mineralization, but long-term (> 45 d) mineralization data were not accurate due to a variety of problems specific to each technique. Buried bags and covered cylinders did not retain mineralized N due to water movement into and out of the containers. Neither resin method captured all of the mineralized N that leached through the soil columns, but the new soil-resin trap method tracked field soil water content better than all other in situ methods evaluated. With further refinement and validation, the new soil-resin trap method may be a useful in situ incubation technique for measuring net N mineralization rates of organic soil amendments.

Prediction of the Dynamic Yield Strength of Metals Using Two Structural-Temporal Parameters

NASA Astrophysics Data System (ADS)

Selyutina, N. S.; Petrov, Yu. V.

2018-02-01

The behavior of the yield strength of steel and a number of aluminum alloys is investigated in a wide range of strain rates, based on the incubation time criterion of yield and the empirical models of Johnson-Cook and Cowper-Symonds. In this paper, expressions for the parameters of the empirical models are derived through the characteristics of the incubation time criterion; a satisfactory agreement of these data and experimental results is obtained. The parameters of the empirical models can depend on some strain rate. The independence of the characteristics of the incubation time criterion of yield from the loading history and their connection with the structural and temporal features of the plastic deformation process give advantage of the approach based on the concept of incubation time with respect to empirical models and an effective and convenient equation for determining the yield strength in a wider range of strain rates.

Kinetic activity, membrane mitochondrial potential, lipid peroxidation, intracellular pH and calcium of frozen/thawed bovine spermatozoa treated with metabolic enhancers.

PubMed

Boni, R; Gallo, A; Cecchini, S

2017-01-01

Owing to the progressive decline of sperm motility during storage there is a need to find substances capable of enhancing sperm energy metabolism and motility and/or preserving it from oxidative damage. The aim of this study was to evaluate in frozen/thawed bovine spermatozoa the effect of several compounds, such as myo-inositol, pentoxifylline, penicillamine + hypotaurine + epinephrine mixture (PHE), caffeine and coenzyme Q10+ zinc + d-aspartate mixture (CZA), on either kinetic or metabolic parameters. Sperm kinetics was evaluated by Sperm Class Analyser whereas specific fluorochromes were used to evaluated mitochondrial membrane potential (MMP), intracellular pH, intracellular calcium concentration and lipid peroxidation. Lipid peroxidation was also evaluated by TBARS analysis. Treatments significantly affected total and progressive motility with different dynamics in relation to the incubation time. After the first hour of incubation, CZA treatment produced the best performance in total and progressive sperm motility as well as in curvilinear velocity, average path velocity and amplitude of head displacement, whereas pentoxifylline stimulated the highest straight-line velocity. MMP showed higher values (p < 0.01) after treatment with pentoxifylline and PHE. Intracytoplasmic calcium concentration and lipid peroxidation were significantly (p < 0.05) affected by the incubation time rather than the treatments. Intracellular pH varied significantly (p < 0.01) in relation to either the incubation time or treatments. In particular, it showed a progressive increase throughout incubation with values in control group significantly higher than in myo-inositol, PHE, caffeine, pentoxifylline and CZA groups (7.37 ± 0.03 vs. 7.29 ± 0.03, 7.28 ± 0.03, 7.26 ± 0.03, 7.22 ± 0.03 and 7.00 ± 0.03, respectively; p < 0.01).; however, among treatments, CZA displayed the lowest values. Significant correlations were found between sperm kinetic and metabolic parameters. These findings provide new comparative information on the effects of putative metabolic enhancers on kinetics and metabolic activities of bovine spermatozoa. In this study, a rapid methodological approach for evaluating sperm quality is proposed. © 2016 American Society of Andrology and European Academy of Andrology.

Importance of analytically verifying chemical treatments

USGS Publications Warehouse

Rach, J.J.; Gaikowski, M.P.; Olson, J.J.

1997-01-01

Hydrogen peroxide is considered a low regulatory priority compound by the U.S. Food and Drug Administration. It is used to control fungal infections on fish eggs. We studied the treatment profiles of hydrogen peroxide in Heath, McDonald egg jar, and Clark-Williamson incubators during treatments intended to deliver an effective regimen of at least 500 ??L hydrogen peroxide/L (i.e., treatments of 500 and 1,000 ??L/L) for 15 min. Hydrogen peroxide concentrations decreased with increasing distance from the influent water in both Heath and Clark-Williamson incubators. The top treatment tray (tray 2) of the Heath incubator received more than 90% of the intended regimen during the 500 ??L/L treatment, whereas at 1,000 ??L/L, all trays had hydrogen peroxide concentrations at or above 500 ??L/L for 15 min. None of the compartments in the Clark-Williamson incubator received the intended therapeutic regimen when treated at 500 ??L/L. The McDonald egg jar system distributed the intended concentration for the designated treatment period in all jars, except those located directly below the influent water. Our results indicate that dilution of therapeutants applied through certain egg incubation systems significantly decreases the efficacy of treatments and may render them ineffective. The dilution characteristics of egg incubation systems should be assessed in order to ensure proper delivery of all intended chemical concentrations and exposure regimens. Suggestions for maintaining the minimum effective concentrations in evaluated incubators are included.

Biological durability and oxidative potential of man-made vitreous fibres as compared to crocidolite asbestos fibres.

PubMed

Hippeli, S; Dornisch, K; Wiethege, T; Gillissen, A; Müller, K M; Elstner, E F

2001-01-01

In this study we investigated relationships between redox properties and biodurability of crocidolite asbestos fibres and three different man-made vitreous fibres (MMVF): traditional stone wool fibres (MMVF 21), glass fibres (MMVF 11) and refractory ceramic fibres (RCF). Each fibre type was incubated up to 22 weeks in four different incubation media: gamble solution (GS) pH 5.0 and pH 7.4, representing blood plasma without proteins, and surfactant-like solution (SLS) pH 5.0 and pH 7.4. During incubation time aliquots of incubation mixtures were removed and analysed in a biochemical model reaction, mimicking activated phagocytes. In addition, changes of fibre morphology and chemical composition were examined using SEM- and EDX-technology. In the presence of crocidolite asbestos fibres and MMVF 21 the formation of OH*-radicals according to the Haber-Weiss sequence could be demonstrated, whereas MMVF 11 and RCF showed no reactivity. Crocidolite asbestos fibres exhibited a significant higher activity compared with the stone wool fibres at the onset of incubation. The oxidative capacities of these fibre types were shown to depend on both specific surface area and iron content. The oxidative potentials of crocidolite asbestos fibres as well as MMVF 21 were not constant during incubation over several weeks in each incubation medium. The reactivities showed sinoidal curves including reactivities much higher than those at the onset of incubation time. These irregular changes of oxidative capacity may be explained by changes of the redox state of fibre surface-complexed iron. Furthermore our results showed clear differences between incubation of fibres in GS and SLS, respectively, indicating that phospholipids play an important part in fibre dissolution behaviour and oxidative reactivity. In conclusion we suggest, that biodurability testing procedures should not exclusively concentrate on dissolution rates of fibres. They should include fibre characteristics concerning known pathogenic mechanisms to evaluate the real toxic potential of the fibre type looking at. Secondly we suggest, that phospholipids should be constituents of incubation liquids used for standardised fibre biodurability test procedures thus representing more realistic incubation conditions.

Trypanosoma cruzi biochemical changes and cell death induced by an organometallic platinum based compound.

PubMed

Mosquillo, M Florencia; Bilbao, Lucía; Hernández, Fabricio; Tissot, Florencia; Gambino, Dinorah; Garat, Beatriz; Pérez-Díaz, Leticia

2018-05-09

Chagas disease is an endemic illness in Latin America caused by the parasite Trypanosoma cruzi. Current chemotherapies are old and inadequate, and the emergence of drug resistant strains underscores the need of new drugs. Platinum-based complexes have been shown to be a promising approach against parasitic diseases. In this work, the effect of 1,1'-bis(diphenylphosphino)ferrocene pyridine-2-thiolate-1-oxide Pt(II) hexafluorophosphate, Pt-dppf-mpo, was studied on T. cruzi. A promising anti-trypanosomal activity was determined for the CL Brener strain with a low cytotoxicity determined using in vitro cultured mammal cells. The compound uptake in parasites treated with concentrations of 1x and 10x the IC 50 value, reached ~75 and 19% respectively. Pt-dppf-mpo induced necrosis after 24h of parasite incubation. This event was preceded by depolarization of mitochondrial membrane potential. Cell vitality assays showed high esterase activity in treated parasites. However, despite this increase in metabolic activity, treated epimastigotes showed rounded morphology and loss of flagellum with a reduction in mobility as compound concentration and/or time of incubation was increased. Finally, we demonstrate that Pt-dppf-mpo incubation also affects the trypomastigote infection process as well as the infection persistence evaluated as the number of amastigotes per cell in a dose dependent manner. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Rapid enzymatic hydrolysis using a novel recombinant β-glucuronidase in benzodiazepine urinalysis.

PubMed

Morris, Ayodele A; Chester, Scot A; Strickland, Erin C; McIntire, Gregory L

2014-10-01

Only trace amounts of parent benzodiazepines are present in urine following extensive metabolism and conjugation. Thus, hydrolysis of glucuronides is necessary for improved detection. Enzyme hydrolysis is preferred to retain identification specificity, but can be costly and time-consuming. The assessment of a novel recombinant β-glucuronidase for rapid hydrolysis in benzodiazepine urinalysis is presented. Glucuronide controls for oxazepam, lorazepam and temazepam were treated with IMCSzyme™ recombinant β-glucuronidase. Hydrolysis efficiency was assessed at 55°C and at room temperature (RT) using the recommended optimum pH. Hydrolysis efficiency for four other benzodiazepines was evaluated solely with positive patient samples. Maximum hydrolysis of glucuronide controls at 5 min at RT (mean analyte recovery ≥ 94% for oxazepam and lorazepam and ≥ 80% for temazepam) was observed. This was considerably faster than the optimized 30 min incubation time for the abalone β-glucuronidase at 65°C. Mean analyte recovery increased at longer incubation times at 55°C for temazepam only. Total analyte in patient samples compared well to targets from abalone hydrolysis after recombinant β-glucuronidase hydrolysis at RT with no incubation. Some matrix effect, differential reactivity, conjugation variability and transformation impacting total analyte recovery were indicated. The unique potential of the IMCSzyme™ recombinant β-glucuronidase was demonstrated with fast benzodiazepine hydrolysis at RT leading to decreased processing time without the need for heat activation. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Studies on the intracellular Ca2+ concentration of thawed dog spermatozoa: influence of Equex from different sources, two thawing diluents and post-thaw incubation in capacitating conditions.

PubMed

Peña, A I; López-Lugilde, L; Barrio, M; Becerra, J J; Quintela, L A; Herradón, P G

2003-02-01

The addition of 0.5% (v/v) of Equex STM Paste (Nova Chemical Sales, Scituate Inc., MA, USA), whose active ingredient is sodium dodecyl sulphate (SDS), to a Tris-egg yolk extender was demonstrated to improve the longevity of frozen-thawed dog spermatozoa during in vitro incubation at 38 degrees C. The aim of the first experiment was to compare the effects of two SDS-containing compounds, Equex STM Paste and Equex Pasta (Minitüb, Tiefenbach, Germany), when added to a Tris-egg yolk based extender, on the post-thaw longevity of dog spermatozoa, as well as on the intracellular Ca2+ concentration of spermatozoa, during post-thaw incubation at 38 degrees C. The post-thaw sperm survival and longevity, as well as the quality of the sperm movement, were significantly better when using Equex STM Paste. Such prolonged sperm longevity, however, was associated to a higher intracellular Ca2+ concentration in a large subpopulation of the live spermatozoa. A second experiment was aimed to evaluate the effects of sperm dilution immediately post-thaw with a Tris buffer containing glucose or fructose. The two Tris buffers were no different for any of the sperm parameters studied. The aim of a third experiment was to evaluate the sperm longevity, motility patterns and intracellular Ca2+ concentration of cryopreserved dog spermatozoa during post-thaw incubation in capacitating conditions [canine capacitating medium (CCM) with or without 5 microg/ml of heparin]. Heparin had no significant effects on any of the sperm parameters evaluated. During the first 8 h of incubation, the majority of the live spermatozoa had a high intracellular Ca2+ content. However, after 8-10 h of incubation, it had significantly declined. The highest proportion of fast motile sperm, and the highest curvilinear velocity, average path velocity and amplitude of lateral head displacement for the total motile sperm were observed during the 2-4-h incubation period. It was concluded that: (a) the addition of 0.5% (v/v) of Equex STM Paste to a Tris-egg yolk based extender significantly improved the post-thaw longevity of dog spermatozoa, but the same concentration of Equex Pasta had no significant beneficial effects; (b) sperm dilution after thawing with a Tris buffer containing glucose or fructose made no difference in post-thaw sperm longevity; (c) the addition of 5 microg/ml of heparin to CCM had no significant capacitating effects on frozen-thawed dog spermatozoa.

Incubation of ethanol reinstatement depends on test conditions and how ethanol consumption is reduced.

PubMed

Ginsburg, Brett C; Lamb, R J

2015-04-01

In reinstatement studies (a common preclinical procedure for studying relapse), incubation occurs (longer abstinence periods result in more responding). This finding is discordant with the clinical literature. Identifying determinants of incubation could aid in interpreting reinstatement and identifying processes involved in relapse. Reinstated responding was examined in rats trained to respond for ethanol and food under a multiple concurrent schedule (Component 1: ethanol FR5, food FR150; Component 2: ethanol FR5, food FR5-alternating across the 30-min session). Ethanol consumption was then reduced for 1 or 16 sessions either by suspending training (rats remained in home cage) or by providing alternative reinforcement (only Component 2 stimuli and contingencies were presented throughout the session). In the next session, stimuli associated with Component 1 were presented and responses recorded but ethanol and food were never delivered. Two test conditions were studied: fixed-ratio completion either produced ethanol- or food-associated stimuli (signaled) or had no programmed consequence (unsignaled). Incubation of ethanol responding was observed only after suspended training during signaled test sessions. Incubation of food responding was also observed after suspended training. These results are most consistent with incubation resulting from a degradation of feedback functions limiting extinction responding, rather than from increased motivation. Copyright © 2015 Elsevier B.V. All rights reserved.

Measles with a possible 23 day incubation period.

PubMed

Fitzgerald, Tove L; Durrheim, David N; Merritt, Tony D; Birch, Christopher; Tran, Thomas

2012-09-30

Measles virus (MV) eradication is biologically, technically and operationally feasible. An essential feature in understanding the chain of MV transmission is its incubation period, that is, the time from infection to the onset of symptoms. This period is important for determining the likely source of infection and directing public health measures to interrupt ongoing transmission. Long measles incubation periods have rarely been documented in the literature. We report on a previously healthy 11-year-old Australian boy who was confirmed with measles genotype D9 infection following travel in the Philippines. Epidemiological evidence supported an unusually long incubation period of at least 23 days and virological evidence was consistent with this finding. Although public health control measures such as post exposure prophylaxis, isolation and surveillance of susceptible individuals should continue to be based on the more common incubation period, a longer incubation period may occasionally explain an unexpected measles case. This work is copyright. Apart from any use as permitted under the Copyright Act 1968, no part may be reproduced by any process without prior written permission from the Commonwealth. Requests and inquiries concerning reproduction and rights should be addressed to the Commonwealth Copyright Administration, Attorney General's Department, Robert Garran Offices, National Circuit, Barton ACT 2600 or posted at http://www.ag.gov.au/cca.

Extensive processing of sediment pore water dissolved organic matter during anoxic incubation as observed by high-field mass spectrometry (FTICR-MS).

PubMed

Valle, Juliana; Gonsior, Michael; Harir, Mourad; Enrich-Prast, Alex; Schmitt-Kopplin, Philippe; Bastviken, David; Conrad, Ralf; Hertkorn, Norbert

2018-02-01

Dissolved organic matter (DOM) contained in lake sediments is a carbon source for many microbial degradation processes, including aerobic and anaerobic mineralization. During anaerobic degradation, DOM is partially consumed and transformed into new molecules while the greenhouse gases methane (CH 4 ) and carbon dioxide (CO 2 ) are produced. In this study, we used ultrahigh resolution mass spectrometry to trace differences in the composition of solid-phase extractable (PPL resin) pore water DOM (SPE-DOM) isolated from surface sediments of three boreal lakes before and after 40 days of anoxic incubation, with concomitant determination of CH 4 and CO 2 evolution. CH 4 and CO 2 production detected by gas chromatography varied considerably among replicates and accounted for fractions of ∼2-4 × 10 -4 of sedimentary organic carbon for CO 2 and ∼0.8-2.4 × 10 -5 for CH 4 . In contrast, the relative changes of key bulk parameters during incubation, such as relative proportions of molecular series, elemental ratios, average mass and unsaturation, were regularly in the percent range (1-3% for compounds decreasing and 4-10% for compounds increasing), i.e. several orders of magnitude higher than mineralization alone. Computation of the average carbon oxidation state in CHO molecules of lake pore water DOM revealed rather non-selective large scale transformations of organic matter during incubation, with depletion of highly oxidized and highly reduced CHO molecules, and formation of rather non-labile fulvic acid type molecules. In general, proportions of CHO compounds slightly decreased. Nearly saturated CHO and CHOS lipid-like substances declined during incubation: these rather commonplace molecules were less specific indicators of lake sediment alteration than the particular compounds, such as certain oxygenated aromatics and carboxyl-rich alicyclic acids (CRAM) found more abundant after incubation. There was a remarkable general increase in many CHNO compounds during incubation across all lakes. Differences in DOM transformation between lakes corresponded with lake size and water residence time. While in the small lake Svarttjärn, CRAM increased during incubation, lignin-and tannin-like compounds were enriched in the large lake Bisen, suggesting selective preservation of these rather non-labile aromatic compounds rather than recent synthesis. SPE-DOM after incubation may represent freshly synthesized compounds, leftover bulk DOM which is primarily composed of intrinsically refractory molecules and/or microbial metabolites which were not consumed in our experiments. In spite of a low fraction of the total DOM being mineralized to CO 2 and CH 4 , the more pronounced change in molecular DOM composition during the incubation indicates that diagenetic modification of organic matter can be substantial compared to complete mineralization. Copyright © 2017 Elsevier Ltd. All rights reserved.

Evaluation of motility, membrane status and DNA integrity of frozen-thawed bottlenose dolphin (Tursiops truncatus) spermatozoa after sex-sorting and recryopreservation.

PubMed

Montano, G A; Kraemer, D C; Love, C C; Robeck, T R; O'Brien, J K

2012-06-01

Artificial insemination (AI) with sex-sorted frozen-thawed spermatozoa has led to enhanced management of ex situ bottlenose dolphin populations. Extended distance of animals from the sorting facility can be overcome by the use of frozen-thawed, sorted and recryopreserved spermatozoa. Although one bottlenose dolphin calf had been born using sexed frozen-thawed spermatozoa derived from frozen semen, a critical evaluation of in vitro sperm quality is needed to justify the routine use of such samples in AI programs. Sperm motility parameters and plasma membrane integrity were influenced by stage of the sex-sorting process, sperm type (non-sorted and sorted) and freezing method (straw and directional) (P<0.05). After recryopreservation, sorted spermatozoa frozen with the directional freezing method maintained higher (P<0.05) motility parameters over a 24-h incubation period compared to spermatozoa frozen using straws. Quality of sperm DNA of non-sorted spermatozoa, as assessed by the sperm chromatin structure assay (SCSA), was high and remained unchanged throughout freeze-thawing and incubation processes. Though a possible interaction between Hoechst 33342 and the SCSA-derived acridine orange was observed in stained and sorted samples, the proportion of sex-sorted, recryopreserved spermatozoa exhibiting denatured DNA was low (6.6±4.1%) at 6 h after the second thawing step and remained unchanged (P>0.05) at 24 h. The viability of sorted spermatozoa was higher (P<0.05) than that of non-sorted spermatozoa across all time points after recryopreservation. Collective results indicate that bottlenose dolphin spermatozoa undergoing cryopreservation, sorting and recryopreservation are of adequate quality for use in AI.

Co-incubation of PMN and CaCo-2 cells modulates inflammatory potential.

PubMed

Schaefer, M B; Schaefer, C A; Hecker, M; Morty, R E; Witzenrath, M; Seeger, W; Mayer, K

2017-05-20

Polymorphonuclear granulocytes (PMN) are activated in inflammatory reactions. Intestinal epithelial cells are relevant for maintaining the intestinal barrier. We examined interactions of PMN and intestinal epithelial cell-like CaCo-2 cells to elucidate their regulation of inflammatory signalling and the impact of cyclooxygenase (COX), nitric oxide (NO) and platelet-activating factor (PAF). Human PMN and CaCo-2 cells, separately and in co-incubation, were stimulated with the calcium ionophore A23187 or with N-Formyl-methionyl-leucyl-phenylalanin (fMLP) that activates PMN only. Human neutrophil elastase (HNE) and respiratory Burst were measured. To evaluate the modulation of inflammatory crosstalk we applied inhibitors of COX (acetyl salicylic acid; ASA), NO-synthase (N-monomethyl-L-arginin; L-NMMA), and the PAF-receptor (WEB2086). Unstimulated, co-incubation of CaCo-2 cells and PMN led to significantly reduced Burst and elevated HNE as compared to PMN. After stimulation with A23187, co-incubation resulted in an inhibition of Burst and HNE. Using fMLP co-incubation failed to modulate Burst but increased HNE. Without stimulation, all three inhibitors abolished the effect of co-incubation on Burst but did not change HNE.  ASA partly prevented modulation of Burst L-NMMA and WEB2086 did not change Burst but abolished mitigation of HNE. Without stimulation, co-incubation reduced Burst and elevated HNE. Activation of PMN and CaCo-2 cells by fMLP as compared to A23187 resulted in a completely different pattern of Burst and HNE, possibly due to single vs. dual cell activation. Anti-inflammatory effect of co-incubation might in part be due to due to COX-signalling governing Burst whereas NO- and PAF-dependent signalling seemed to control HNE release.

Metabolic effects of photodynamically induced apoptosis in an erythroleukemic cell line. A (31)P NMR spectroscopic study of Victoria-Blue-BO-sensitized TF-1 cells.

PubMed

Viola, A; Lutz, N W; Maroc, C; Chabannon, C; Julliard, M; Cozzone, P J

2000-03-01

Victoria Blue BO (VB BO) is a new and promising photosensitizer currently being evaluated for photodynamic therapy (PDT). Its photochemical processes are mediated by oxygen radicals, but do not involve singlet oxygen. We used (31)P NMR spectroscopy of VB-BO sensitized TF-1 leukemic cells to gain further insight into the biochemical mechanisms underlying PDT-induced cell death. Sham-treatment experiments were performed to evaluate the effects of this photosensitizer in the absence of light irradiation. Significant metabolic differences were detected for TF-1 cells incubated with VB BO but not exposed to light, as compared with native cells (controls). These changes include reductions in phosphocreatine, UDP-hexose and phosphodiester levels (as percentage of total phosphate) and slightly reduced intracellular pH. Complete phosphocreatine depletion, significant acidification and concomitant inorganic-phosphate accumulation were observed for TF-1 cells irradiated after incubation with VB BO. Moreover, significant changes in phospholipid metabolites, i.e., accumulation of cytidine 5'-diphosphate choline and a decrease in phosphodiester levels, were observed for PDT-treated vs. sham-treated cells. Perturbations of phospholipid metabolism may be involved in programmed cell death, and the detection of a characteristic DNA ladder pattern by gel electrophoresis confirmed the existence of apoptosis in PDT-treated TF-1 cells. Copyright 2000 Wiley-Liss, Inc.

Evaluating the effectiveness of a Mexican strain of Duddingtonia flagrans as a biological control agent against gastrointestinal nematodes in goat faeces.

PubMed

Ojeda-Robertos, N F; Mendoza-de Gives, P; Torres-Acosta, J F J; Rodríguez-Vivas, R I; Aguilar-Caballero, A J

2005-06-01

The use of Duddingtonia flagrans in the control of goat nematodes was investigated. Initially, the time of passage of chlamydospores through the digestive tract of goats was evaluated. Two groups of seven parasite-free kids were formed. Group A received a single dose of 3.5x10(6) D. flagrans chlamydospores (FTHO-8 strain) per kg of live weight. Group B did not receive any chlamydospores. Faeces were obtained from each kid daily from day 4 prior to inoculation until day 5 post-inoculation (PI) and were placed in Petri dishes containing water agar. Gastrointestinal nematode infective larvae were added to each Petri dish and incubated at 25 degrees C for 7 days. Petri dishes were examined to detect the fungus and trapped nematodes. A second trial evaluated the effect of D. flagrans on the number of gastrointestinal nematode larvae harvested from goat faecal cultures in naturally infected goats. Two groups of seven goats were formed. The treated group received a single dose of 3.5x10(6) D. flagrans chlamydospores per kg of liveweight. The control group did not receive any chlamydospores. Faeces were obtained twice daily from each kid. Two faecal cultures were made for each kid. One was incubated for 7 days and the other for 14 days. Gastrointestinal nematode larvae were recovered from each culture and counted. Percentage of larval development reduction was determined using a ratio of larvae/eggs deposited in the control and treated groups. Duddingtonia flagrans survived the digestive process of goats, and maintained its predatory activity, being observed from 21 to 81 h PI (3 to 4 days). A reduction in the infective larvae population in the treated group compared to the non-treated group was observed in both incubation periods (7 days: 5.3-36.0%; 14 days: 0-52.8%, P>0.05). Although a single inoculation of D. flagrans can induce a reduction of infective larvae collected from faeces, a different scheme of dosing may be needed to enhance the efficacy of D. flagrans in goats.

Effects of modification of in vitro fertilization techniques on the sex ratio of the resultant bovine embryos.

PubMed

Iwata, H; Shiono, H; Kon, Y; Matsubara, K; Kimura, K; Kuwayama, T; Monji, Y

2008-05-01

The duration of sperm-oocyte co-incubation has been observed to affect the sex ratio of in vitro produced bovine embryos. The purpose of this study was to investigate some factors that may be responsible for the skewed sex ratio. The factors studied were selected combinations of the duration of co-incubation, the presence or absence of cumulus cells, and the level of hyaluronic acid (HA) in the culture medium. Experiment 1 examined the effect of selected combinations of different factors during the fertilization phase of in vitro oocyte culture. The factors were the nature of the sperm or its treatment, the duration of the sperm-oocyte co-incubation, and the level of hyaluronic acid in the culture medium. In experiment 2, the capacitation of frozen-thawed-Percoll-washed sperm (control), pre-incubated, and non-binding sperm was evaluated by the zona pellucida (ZP) binding assay and the hypo-osmotic swelling test (HOST). The purpose of experiment 3 was to determine the oocyte cleavage rate and sex ratio of the embryos (>5 cells) produced as a consequence of the 10 treatments used in experiment 1. In treatments 1-3 (experiments 1 and 3) COC were co-cultured with sperm for 1, 5 or 18 h. Polyspermic fertilization rose as the co-incubation period increased (1 h 6.5%, 5 h 15.9%, 18 h 41.8%; P<0.05), and the highest rate of normal fertilization was observed for 5h culture (73.4%; P<0.05). The sex ratio was significantly (P<0.05) skewed from the expected 50:50 towards males following 1 h (64.4%) and 5 h (67.3%) co-incubation, but was not affected by 18 h incubation (52.3%). In treatment 4, sperm was pre-incubated for 1h and cultured with COC for 5 h. Relative to control sperm, pre-incubation of sperm increased ZP binding (116 versus 180 per ZP; P<0.05) and decreased the proportion of HOST positive sperm (65.8-48.6%; P<0.05; experiment 2). Pre-incubation did not affect the rates of polyspermy, normal fertilization or the sex ratio of the embryos (experiments 1 and 3). The oocytes used in treatments 5-10 of experiments 1 and 3 were denuded prior to fertilization. Co-incubation of denuded oocytes for 1h (treatment 5) or 5h (treatment 6) resulted in levels of polyspermic fertilization similar to that for treatment 2 with significantly lower levels of normal fertilization (41.7% and 52.6%, respectively; P<0.05), and the 1h co-incubation significantly skewed (P<0.05) the proportion of male embryos to 70.0%. Denuded oocytes were fertilized for 5h with sperm unable to bind to cumulus cells (NB sperm) in treatment 7 or those that bound to cumulus cells (B) in treatment 8. These two treatments had similar rates of polyspermic, normal and non-fertilization. However, the B sperm caused the sex ratio of the embryos to be significantly skewed to males (63.9%; P<0.05). Fertilization of denuded oocytes in medium containing hyaluronic acid (0.1 mg/ml, treatment 9; 1.0 mg/ml treatment 10) significantly (P<0.05) reduced the incidence of polyspermic fertilization relative to treatments 2 and 6, and normal fertilization relative to treatment 2, but did not affect the sex ratio of the embryos. It was concluded that exposure of sperm to cumulus cells, either before fertilization of denuded oocytes or during the process of fertilization of complete COC, increased the proportion of male embryos produced by in vitro culture. It was hypothesized that this may be due to the capacitation state of the sperm, the cumulus-sperm interaction, and/or the ability of the sperm to bind to cumulus cells or oocytes.

Periods of cardiovascular susceptibility to hypoxia in embryonic american alligators (Alligator mississippiensis)

PubMed Central

Tate, Kevin B.; Rhen, Turk; Eme, John; Kohl, Zachary F.; Crossley, Janna; Elsey, Ruth M.

2016-01-01

During embryonic development, environmental perturbations can affect organisms' developing phenotype, a process known as developmental plasticity. Resulting phenotypic changes can occur during discrete, critical windows of development. Critical windows are periods when developing embryos are most susceptible to these perturbations. We have previously documented that hypoxia reduces embryo size and increases relative heart mass in American alligator, and this study identified critical windows when hypoxia altered morphological, cardiovascular function and cardiac gene expression of alligator embryos. We hypothesized that incubation in hypoxia (10% O2) would increase relative cardiac size due to cardiac enlargement rather than suppression of somatic growth. We exposed alligator embryos to hypoxia during discrete incubation periods to target windows where the embryonic phenotype is altered. Hypoxia affected heart growth between 20 and 40% of embryonic incubation, whereas somatic growth was affected between 70 and 90% of incubation. Arterial pressure was depressed by hypoxic exposure during 50–70% of incubation, whereas heart rate was depressed in embryos exposed to hypoxia during a period spanning 70–90% of incubation. Expression of Vegf and PdgfB was increased in certain hypoxia-exposed embryo treatment groups, and hypoxia toward the end of incubation altered β-adrenergic tone for arterial pressure and heart rate. It is well known that hypoxia exposure can alter embryonic development, and in the present study, we have identified brief, discrete windows that alter the morphology, cardiovascular physiology, and gene expression in embryonic American alligator. PMID:27101296

Optimisation of low temperature extraction of banana juice using commercial pectinase.

PubMed

Sagu, Sorel Tchewonpi; Nso, Emmanuel Jong; Karmakar, Sankha; De, Sirshendu

2014-05-15

The objective of this work was to develop a process with optimum conditions for banana juice. The procedure involves hydrolyzing the banana pulp by commercial pectinase followed by cloth filtration. Response surface methodology with Doehlert design was utilised to optimize the process parameters. The temperature of incubation (30-60 °C), time of reaction (20-120 min) and concentration of pectinase (0.01-0.05% v/w) were the independent variables and viscosity, clarity, alcohol insoluble solids (AIS), total polyphenol and protein concentration were the responses. Total soluble sugar, pH, conductivity, calcium, sodium and potassium concentration in the juice were also evaluated. The results showed reduction of AIS and viscosity with reaction time and pectinase concentration and reduction of polyphenol and protein concentration with temperature. Using numerical optimization, the optimum conditions for the enzymatic extraction of banana juice were estimated. Depectinization kinetics was also studied at optimum temperature and variation of kinetic constants with enzyme dose was evaluated. Copyright © 2013 Elsevier Ltd. All rights reserved.

Chain response of microbial loop to the decay of a diatom bloom in the East China Sea

NASA Astrophysics Data System (ADS)

Wu, Linnan; Lin, Shiquan; Huang, Lingfeng; Lu, Jiachang; Chen, Wenzhao; Guo, Weidong; Zhang, Wuchang; Xiao, Tian; Sun, Jun

2016-02-01

Algal bloom has been regarded as one of the key causes for the summer hypoxia phenomena in the bottom water adjacent to the Yangtze River estuary in the East China Sea. Although a series of biological processes within microbial loop are involved in the development of oxygen depletion during the bloom decay, little has been known about the dynamics of microorganisms in response to the decaying process of the bloom through trophic interaction context. Here, we report some preliminary results of our observations about the response of microbial loop to the bloom decay, based on the onboard incubation experiments for 10 days during a diatom bloom near the Yangtze River estuary in August, 2011. Light and dark incubations were conducted to simulate the bloom decay inside and below the euphotic layer, respectively. In the first stage of bloom decay (Day 0 to Day 4), rapid response was found in heterotrophic bacteria (HB) and ciliate growth, which was in accordance with the decrease of total Chl a, indicating a "bottom-up" control at the early stage of bloom decay. However, the increase of heterotrophic nanoflagellates (HNF) abundance was rather inconspicuous, suggesting predation pressure on HNF from ciliate or other predator at this stage. In the second stage (Day 4 to Day 8), HB and ciliate decreased rapidly with the increase of HNF, revealing the release of HNF form ciliate predation, which suggested a "top-down" control. In the last stage of our experiment (Day 8 to Day 10), the trophic interactions were more complex, but it also implied a "top-down" control within the microbial loop. Meanwhile, virus had been monitored in the whole process of our incubations. It was found that virus lysed microalgae at the first stage, and lysed HB at the second stage. In addition, the bacterial mortality was principally caused by HNF grazing in the light-sufficient incubations and by viral lysis in the light-insufficient incubations. Our results suggest tight trophic interactions within the microbial loop in the decaying process of the algal bloom, which may assist our understanding of the role of microbial loop in hypoxia formation in coastal waters.

Process of Infection with Bacteriophage φX174

PubMed Central

Dalgarno, L.; Sinsheimer, Robert L.

1968-01-01

A group of temperature-sensitive mutants of φX174 has been isolated which can go through a single, normal one-step growth cycle at 40 C but fail to form plaques at this temperature. Such mutants fail to initiate a second cycle at 40 C; however they can gain the capacity to infect at 40 C, upon incubation for 10 min in broth at 30 C. In regaining the ability to infect, the phage appear to undergo a temperature-dependent conformational alteration. The inverse process, a reversible loss of ability to infect at 40 C, is observed when such phage produced at 30 C are incubated for 2 hr at 40 C. The defect in initiation of a second cycle of infection appears to be in the injection of viral deoxyribonucleic acid. A two-step complementation test has been used to identify the cistron coding for the affected function. Such mutants are also unusually sensitive to an irreversible thermal inactivation when incubated at 40 C. PMID:4883013

Dissolved Organic Carbon: Nitrate Ratios as a Driver of Methane Fluxes in Stream Ecosystems

NASA Astrophysics Data System (ADS)

Sullivan, B. W.; Wymore, A.; Schade, J. D.; McDowell, W. H.

2016-12-01

Fluvial ecosystems are poorly understood components of the global methane (CH4) budget because the ecology of CH4 fluxes in streams has yet to be sufficiently elucidated. Both CH4 production and uptake via oxidation are microbially mediated processes, but it is unclear where in the fluvial environment are the sources and sinks of CH4 and what role terrestrial inputs of carbon (C) and nutrients have on the magnitude and direction of CH4 flux. To address these uncertainties, we measured CH4 fluxes in a laboratory incubation from two temperate headwater streams that differed in ambient dissolved organic carbon (DOC) and nitrate (NO3-) concentrations. We amended stream water and sediment microcosms from each site with labile DOC from senesced leaf litter to assess how DOC concentration and the DOC:NO3- ratio affect proximate controls on CH4 flux. Lastly, we manipulated sediment and water column ratios (0-100%) to estimate sources and fates of CH4 flux within the ecosystem. We measured CH4 fluxes for the first 120 minutes of the incubation to simulate short-term, in stream processes. Initially, streams were a source of methane, but switched to a sink within 120 minutes. Methane fluxes were statistically similar in both stream sediment and water, suggesting that microbial processing of CH4 has similar directionality and magnitude in each environment. Both CH4 oxidation and production were significantly correlated with the DOC: NO3- ratio over the course of the incubation. Early in the incubation, increasing DOC: NO3- increased CH4 flux, but late in the incubation, increasing DOC: NO3- increased CH4 oxidation. Together, our results challenge existing paradigms of CH4 flux in the fluvial environment and identify the DOC:NO3- ratio as a possible mechanism that can explain spatial and temporal CH4 flux patterns in streams.

The Development of a Microbial Challenge Test with Acholeplasma laidlawii To Rate Mycoplasma-Retentive Filters by Filter Manufacturers.

PubMed

Folmsbee, Martha; Lentine, Kerry Roche; Wright, Christine; Haake, Gerhard; Mcburnie, Leesa; Ashtekar, Dilip; Beck, Brian; Hutchison, Nick; Okhio-Seaman, Laura; Potts, Barbara; Pawar, Vinayak; Windsor, Helena

2014-01-01

Mycoplasma are bacteria that can penetrate 0.2 and 0.22 μm rated sterilizing-grade filters and even some 0.1 μm rated filters. Primary applications for mycoplasma filtration include large scale mammalian and bacterial cell culture media and serum filtration. The Parenteral Drug Association recognized the absence of standard industry test parameters for testing and classifying 0.1 μm rated filters for mycoplasma clearance and formed a task force to formulate consensus test parameters. The task force established some test parameters by common agreement, based upon general industry practices, without the need for additional testing. However, the culture medium and incubation conditions, for generating test mycoplasma cells, varied from filter company to filter company and was recognized as a serious gap by the task force. Standardization of the culture medium and incubation conditions required collaborative testing in both commercial filter company laboratories and in an Independent laboratory (Table I). The use of consensus test parameters will facilitate the ultimate cross-industry goal of standardization of 0.1 μm filter claims for mycoplasma clearance. However, it is still important to recognize filter performance will depend on the actual conditions of use. Therefore end users should consider, using a risk-based approach, whether process-specific evaluation of filter performance may be warranted for their application. Mycoplasma are small bacteria that have the ability to penetrate sterilizing-grade filters. Filtration of large-scale mammalian and bacterial cell culture media is an example of an industry process where effective filtration of mycoplasma is required. The Parenteral Drug Association recognized the absence of industry standard test parameters for evaluating mycoplasma clearance filters by filter manufacturers and formed a task force to formulate such a consensus among manufacturers. The use of standardized test parameters by filter manufacturers, including the preparation of the culture broth, will facilitate the end user's evaluation of the mycoplasma clearance claims provided by filter vendors. However, it is still important to recognize filter performance will depend on the actual conditions of use; therefore end users should consider, using a risk-based approach, whether process-specific evaluation of filter performance may be warranted for their application. © PDA, Inc. 2014.

Use of dissolved H2 concentrations to determine distribution of microbially catalyzed redox reactions in anoxic groundwater

USGS Publications Warehouse

Lovley, D.R.; Chapelle, F.H.; Woodward, J.C.

1994-01-01

The potential for using concentrations of dissolved H2 to determine the distribution of redox processes in anoxic groundwaters was evaluated. In pristine aquifers in which standard geochemical measurements indicated that Fe-(III) reduction, sulfate reduction, or methanogenesis was the terminal electron accepting process (TEAP), the H2 concentrations were similar to the H2 concentrations that have previously been reported for aquatic sediments with the same TEAPs. In two aquifers contaminated with petroleum products, it was impossible with standard geochemical analyses to determine which TEAPs predominated in specific locations. However, the TEAPs predicted from measurements of dissolved H2 were the same as those determined directly through measurements of microbial processes in incubated aquifer material. These results suggest that H2 concentrations may be a useful tool for analyzing the redox chemistry of nonequilibrium groundwaters.

Quick counting method for estimating the number of viable microbes on food and food processing equipment.

PubMed

Winter, F H; York, G K; el-Nakhal, H

1971-07-01

A rapid method for estimating the extent of microbial contamination on food and on food processing equipment is described. Microbial cells are rinsed from food or swab samples with sterile diluent and concentrated on the surface of membrane filters. The filters are incubated on a suitable bacteriological medium for 4 hr at 30 C, heated at 105 C for 5 min, and stained. The membranes are then dried at 60 C for 15 min, rendered transparent with immersion oil, and examined microscopically. Data obtained by the rapid method were compared with counts of the same samples determined by the standard plate count method. Over 60 comparisons resulted in a correlation coefficient of 0.906. Because the rapid technique can provide reliable microbiological count information in extremely short times, it can be a most useful tool in the routine evaluation of microbial contamination of food processing facilities and for some foods.

The presence of Enterococcus, coliforms and E. coli in a commercial yeast manufacturing process.

PubMed

O'Brien, S S; Lindsay, D; von Holy, A

2004-07-01

This study evaluated a typical commercial yeast manufacturing process for bacterial contamination. Product line samples of a commercial yeast manufacturing process and the corresponding seed yeast manufacturing process were obtained upstream from the final compressed and dry yeast products. All samples were analysed before (non-PI) and after preliminary incubation (PI) at 37 degrees C for 24 h. The PI procedure was incorporated for amplification of bacterial counts below the lower detection limit. Enterococcus, coliform and Escherichia coli counts were quantified by standard pour-plate techniques using selective media. Presence at all stages and progressive increases in counts of Enterococcus, coliforms and E. coli during processing in the commercial manufacturing operation suggested that the primary source of contamination of both compressed and dry yeast with these bacteria was the seed yeast manufacturing process and that contamination was amplified throughout the commercial yeast manufacturing process. This was confirmed by surveys of the seed yeast manufacturing process which indicated that contamination of the seed yeast with Enterococcus, coliforms and E. coli occurred during scale up of seed yeast biomass destined as inoculum for the commercial fermentation.

Estimation of the incubation period of influenza A (H1N1-2009) among imported cases: addressing censoring using outbreak data at the origin of importation.

PubMed

Nishiura, Hiroshi; Inaba, Hisashi

2011-03-07

Empirical estimates of the incubation period of influenza A (H1N1-2009) have been limited. We estimated the incubation period among confirmed imported cases who traveled to Japan from Hawaii during the early phase of the 2009 pandemic (n=72). We addressed censoring and employed an infection-age structured argument to explicitly model the daily frequency of illness onset after departure. We assumed uniform and exponential distributions for the frequency of exposure in Hawaii, and the hazard rate of infection for the latter assumption was retrieved, in Hawaii, from local outbreak data. The maximum likelihood estimates of the median incubation period range from 1.43 to 1.64 days according to different modeling assumptions, consistent with a published estimate based on a New York school outbreak. The likelihood values of the different modeling assumptions do not differ greatly from each other, although models with the exponential assumption yield slightly shorter incubation periods than those with the uniform exposure assumption. Differences between our proposed approach and a published method for doubly interval-censored analysis highlight the importance of accounting for the dependence of the frequency of exposure on the survival function of incubating individuals among imported cases. A truncation of the density function of the incubation period due to an absence of illness onset during the exposure period also needs to be considered. When the data generating process is similar to that among imported cases, and when the incubation period is close to or shorter than the length of exposure, accounting for these aspects is critical for long exposure times. Copyright © 2010 Elsevier Ltd. All rights reserved.

Multi-layer thermoelectric-temperature-mapping microbial incubator designed for geo-biochemistry applications.

PubMed

Wu, Jin-Gen; Liu, Man-Chi; Tsai, Ming-Fei; Yu, Wei-Shun; Chen, Jian-Zhang; Cheng, I-Chun; Lin, Pei-Chun

2012-04-01

We demonstrate a novel, vertical temperature-mapping incubator utilizing eight layers of thermoelectric (TE) modules mounted around a test tube. The temperature at each layer of the TE module is individually controlled to simulate the vertical temperature profile of geo-temperature variations with depth. Owing to the constraint of non-intrusion to the filled geo-samples, the temperature on the tube wall is adopted for measurement feedback. The design considerations for the incubator include spatial arrangement of the energy transfer mechanism, heating capacity of the TE modules, minimum required sample amount for follow-up instrumental or chemical analysis, and the constraint of non-intrusion to the geo-samples during incubation. The performance of the incubator is experimentally evaluated with two tube conditions and under four preset temperature profiles. Test tubes are either empty or filled with quartz sand, which has comparable thermal properties to the materials in the geo-environment. The applied temperature profiles include uniform, constant temperature gradient, monotonic-increasing parabolic, and parabolic. The temperature on the tube wall can be controlled between 20 °C and 90 °C with an averaged root mean squared error of 1 °C. © 2012 American Institute of Physics

Development and Characterization of Polymer Eco-Composites Based on Natural Rubber Reinforced with Natural Fibers.

PubMed

Stelescu, Maria-Daniela; Manaila, Elena; Craciun, Gabriela; Chirila, Corina

2017-07-11

Natural rubber composites filled with short natural fibers (flax and sawdust) were prepared by blending procedure and the elastomer cross-linking was carried out using benzoyl peroxide. The microbial degradation of composites was carried out by incubating with Aspergillus niger recognized for the ability to grow and degrade a broad range of substrates. The extent of biodegradation was evaluated by weight loss and cross-linking degree study of composites after 2 months incubation in pure shake culture conditions. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR) have proved to be precious and valuable instruments for morphological as well as structural characterization of the composites before and after incubation with Aspergillus niger .

Development and Characterization of Polymer Eco-Composites Based on Natural Rubber Reinforced with Natural Fibers

PubMed Central

Stelescu, Maria-Daniela; Manaila, Elena; Craciun, Gabriela; Chirila, Corina

2017-01-01

Natural rubber composites filled with short natural fibers (flax and sawdust) were prepared by blending procedure and the elastomer cross-linking was carried out using benzoyl peroxide. The microbial degradation of composites was carried out by incubating with Aspergillus niger recognized for the ability to grow and degrade a broad range of substrates. The extent of biodegradation was evaluated by weight loss and cross-linking degree study of composites after 2 months incubation in pure shake culture conditions. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR) have proved to be precious and valuable instruments for morphological as well as structural characterization of the composites before and after incubation with Aspergillus niger. PMID:28773145

Controlling attachment and growth of Listeria monocytogenes in polyvinyl chloride model floor drains using a peroxide chemical, chitosan-arginine, or heat.

PubMed

Berrang, Mark E; Hofacre, Charles L; Frank, Joseph F

2014-12-01

Listeria monocytogenes can colonize a poultry processing plant as a resident in floor drains. Limiting growth and attachment to drain surfaces may help lessen the potential for cross-contamination of product. The objective of this study was to compare a hydrogen peroxide-peroxyacetic acid-based chemical to chitosan-arginine or heat to prevent attachment of or destroy existing L. monocytogenes on the inner surface of model floor drains. L. monocytogenes was introduced to result in about 10(9) planktonic and attached cells within untreated polyvinyl chloride model drain pipes. Treatments (0.13 % peroxide-based sanitizer, 0.1 % chitosan-arginine, or 15 s of hot water at 95 to 100°C) were applied immediately after inoculation or after 24 h of incubation. Following treatment, all pipes were incubated for an additional 24 h; planktonic and attached cells were enumerated by plate count. All treatments significantly (P < 0.05) lowered numbers of planktonic and attached cells recovered. Chitosan-arginine resulted in approximately a 6-log reduction in planktonic cells when applied prior to incubation and a 3-log reduction after the inoculum had a chance to grow. Both heat and peroxide significantly outperformed chitosan-arginine (8- to 9-log reduction) and were equally effective before and after incubation. Heat was the only treatment that eliminated planktonic L. monocytogenes. All treatments were less effective against attached cells. Chitosan-arginine provided about a 4.5-log decrease in attached cells when applied before incubation and no significant decrease when applied after growth. Like with planktonic cells, peroxide-peroxyacetic acid and heat were equally effective before or after incubation, causing decreases ranging from 7 to 8.5 log for attached L. monocytogenes. Applied at the most efficacious time, any of these techniques may lessen the potential for L. monocytogenes to remain as a long-term resident in processing plant floor drains.

Effects of El Niño-driven changes in wind patterns on North Pacific albatrosses.

PubMed

Thorne, L H; Conners, M G; Hazen, E L; Bograd, S J; Antolos, M; Costa, D P; Shaffer, S A

2016-06-01

Changes to patterns of wind and ocean currents are tightly linked to climate change and have important implications for cost of travel and energy budgets in marine vertebrates. We evaluated how El Niño-Southern Oscillation (ENSO)-driven wind patterns affected breeding Laysan and black-footed albatross across a decade of study. Owing to latitudinal variation in wind patterns, wind speed differed between habitat used during incubation and brooding; during La Niña conditions, wind speeds were lower in incubating Laysan (though not black-footed) albatross habitat, but higher in habitats used by brooding albatrosses. Incubating Laysan albatrosses benefited from increased wind speeds during El Niño conditions, showing increased travel speeds and mass gained during foraging trips. However, brooding albatrosses did not benefit from stronger winds during La Niña conditions, instead experiencing stronger cumulative headwinds and a smaller proportion of trips in tailwinds. Increased travel costs during brooding may contribute to the lower reproductive success observed in La Niña conditions. Furthermore, benefits of stronger winds in incubating habitat may explain the higher reproductive success of Laysan albatross during El Niño conditions. Our findings highlight the importance of considering habitat accessibility and cost of travel when evaluating the impacts of climate-driven habitat change on marine predators. © 2016 The Author(s).

Effects of El Niño-driven changes in wind patterns on North Pacific albatrosses

PubMed Central

Thorne, L. H.; Conners, M. G.; Hazen, E. L.; Bograd, S. J.; Antolos, M.; Costa, D. P.; Shaffer, S. A.

2016-01-01

Changes to patterns of wind and ocean currents are tightly linked to climate change and have important implications for cost of travel and energy budgets in marine vertebrates. We evaluated how El Niño-Southern Oscillation (ENSO)-driven wind patterns affected breeding Laysan and black-footed albatross across a decade of study. Owing to latitudinal variation in wind patterns, wind speed differed between habitat used during incubation and brooding; during La Niña conditions, wind speeds were lower in incubating Laysan (though not black-footed) albatross habitat, but higher in habitats used by brooding albatrosses. Incubating Laysan albatrosses benefited from increased wind speeds during El Niño conditions, showing increased travel speeds and mass gained during foraging trips. However, brooding albatrosses did not benefit from stronger winds during La Niña conditions, instead experiencing stronger cumulative headwinds and a smaller proportion of trips in tailwinds. Increased travel costs during brooding may contribute to the lower reproductive success observed in La Niña conditions. Furthermore, benefits of stronger winds in incubating habitat may explain the higher reproductive success of Laysan albatross during El Niño conditions. Our findings highlight the importance of considering habitat accessibility and cost of travel when evaluating the impacts of climate-driven habitat change on marine predators. PMID:27278360

Reciprocal interaction between dental alloy biocorrosion and Streptococcus mutans virulent gene expression.

PubMed

Zhang, Songmei; Qiu, Jing; Ren, Yanfang; Yu, Weiqiang; Zhang, Fuqiang; Liu, Xiuxin

2016-04-01

Corrosion of dental alloys is a major concern in dental restorations. Streptococcus mutans reduces the pH in oral cavity and induces demineralization of the enamel as well as corrosion of restorative dental materials. The rough surfaces of dental alloys induced by corrosion enhance the subsequent accumulation of plaque. In this study, the corrosion process of nickel-chromium (Ni-Cr) and cobalt-chromium (Co-Cr) alloys in a nutrient-rich medium containing S. mutans was studied using inductively coupled plasma atomic emission spectrometry (ICP-AES), X-ray photoelectron spectroscopy (XPS) and electrochemical corrosion test. Our results showed that the release of Ni and Co ions increased, particularly after incubation for 3 days. The electrochemical corrosion results showed a significant decrease in the corrosion resistance (Rp) value after the alloys were immersed in the media containing S. mutans for 3 days. Correspondingly, XPS revealed a reduction in the relative dominance of Ni, Co, and Cr in the surface oxides after the alloys were immersed in the S. mutans culture. After removal of the biofilm, the pre-corroded alloys were re-incubated in S. mutans medium, and the expressions of genes associated with the adhesion and acidogenesis of S. mutans, including gtfBCD, gbpB, fif and ldh, were evaluated by detecting the mRNA levels using real-time reverse transcription polymerase chain reaction (RT-PCR). We found that the gtfBCD, gbpB, ftf and Idh expression of S. mutans were noticeably increased after incubation with pre-corroded alloys for 24 h. This study demonstrated that S. mutans enhanced the corrosion behavior of the dental alloys, on the other hand, the presence of corroded alloy surfaces up-regulated the virulent gene expression in S. mutans. Compared with smooth surfaces, the rough corroded surfaces of dental alloys accelerated the bacteria-adhesion and corrosion process by changing the virulence gene expression of S. mutans.

Simple SPION Incubation as an Efficient Intracellular Labeling Method for Tracking Neural Progenitor Cells Using MRI

PubMed Central

D. M., Jayaseema; Lai, Jiann-Shiun; Hueng, Dueng-Yuan; Chang, Chen

2013-01-01

Cellular magnetic resonance imaging (MRI) has been well-established for tracking neural progenitor cells (NPC). Superparamagnetic iron oxide nanoparticles (SPIONs) approved for clinical application are the most common agents used for labeling. Conventionally, transfection agents (TAs) were added with SPIONs to facilitate cell labeling because SPIONs in the native unmodified form were deemed inefficient for intracellular labeling. However, compelling evidence also shows that simple SPION incubation is not invariably ineffective. The labeling efficiency can be improved by prolonged incubation and elevated iron doses. The goal of the present study was to establish simple SPION incubation as an efficient intracellular labeling method. To this end, NPCs derived from the neonatal subventricular zone were incubated with SPIONs (Feridex®) and then evaluated in vitro with regard to the labeling efficiency and biological functions. The results showed that, following 48 hours of incubation at 75 µg/ml, nearly all NPCs exhibited visible SPION intake. Evidence from light microscopy, electron microscopy, chemical analysis, and magnetic resonance imaging confirmed the effectiveness of the labeling. Additionally, biological assays showed that the labeled NPCs exhibited unaffected viability, oxidative stress, apoptosis and differentiation. In the demonstrated in vivo cellular MRI experiment, the hypointensities representing the SPION labeled NPCs remained observable throughout the entire tracking period. The findings indicate that simple SPION incubation without the addition of TAs is an efficient intracellular magnetic labeling method. This simple approach may be considered as an alternative approach to the mainstream labeling method that involves the use of TAs. PMID:23468856

Leaching of two fungicides in spent mushroom substrate amended soil: Influence of amendment rate, fungicide ageing and flow condition.

PubMed

Álvarez-Martín, Alba; Sánchez-Martín, María J; Ordax, José M; Marín-Benito, Jesús M; Sonia Rodríguez-Cruz, M

2017-04-15

A study has been conducted on the leaching of two fungicides, tebuconazole and cymoxanil, in a soil amended with spent mushroom substrate (SMS), with an evaluation of how different factors influence this process. The objective was based on the potential use of SMS as a biosorbent for immobilizing pesticides in vulnerable soils, and the need to know how it could affect the subsequent transport of these retained compounds. Breakthrough curves (BTCs) for 14 C-fungicides, non-incubated and incubated over 30days, were obtained in columns packed with an unamended soil (S), and this soil amended with SMS at rates of 5% (S+SMS5) and 50% (S+SMS50) under saturated and saturated-unsaturated flows. The highest leaching of tebuconazole (>50% of the total 14 C added) was found in S when a saturated water flow was applied to the column, but the percentage of leached fungicide decreased when a saturated-unsaturated flow was applied in both SMS-amended soils. Also a significant decrease in leaching was observed for tebuconazole after incubation in the column, especially in S+SMS50 when both flows were applied. Furthermore, cymoxanil leaching was complete in S and S+SMS when a saturated flow was applied, and maximum peak concentrations were reached at 1pore volume (PV), although BTCs showed peaks with lower concentrations in S+SMS. The amounts of cymoxanil retained only increased in S+SMS when a saturated-unsaturated flow was applied. A more relevant effect of SMS for reducing the leaching of fungicide was observed when cymoxanil was previously incubated in the column, although mineralization was enhanced in this case. These results are of interest for extending SMS application on the control of the leaching of fungicides with different physicochemical characteristics after different ageing times in the soil and water flow conditions applied. Copyright © 2017 Elsevier B.V. All rights reserved.

Macrophage-derived microvesicles promote proliferation and migration of Schwann cell on peripheral nerve repair

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhan, Chuan, E-mail: zhchuansy@163.com; Ma, Cheng-bin; Yuan, Hong-mou

Background: Macrophages have been implicated in peripheral nerve regeneration. However, whether macrophages-derived microvesicles (MVs) are involved in this process remains unknown. In the present study, the effects of macrophages-derived MVs on proliferation and migration of Schwann cells (SCs) were evaluated in both in vitro and in vivo. Methods: Human monocytic leukaemia cell line (THP-1) was successfully driven to M1 and M2 phenotypes by delivery of either IFN-γ or IL-4, respectively. SCs incubated with M1 or M2 macrophages-derived MVs, the cell migration and proliferation were assessed, and expression levels of nerve growth factor (NGF) and Laminin were measured. A rat model of sciaticmore » nerve was established and the effects of macrophages-derived MVs on nerve regeneration were investigated. Results: M2-derived MVs elevated migration, proliferation, NFG and Laminin protein levels of SCs compared with M1-or M0-derived MVs. The relative expression levels of miR-223 were also increased in M2 macrophages and M2-derived MVs. Transfected M2 macrophages with miR-223 inhibitor then co-incubated with SCs, an inhibition of cell migration and proliferation and a down-regulated levels of NFG and Laminin protein expression were observed. In vivo, M2-derived MVs significantly increased the infiltration and axon number of SCs. Conclusion: M2-derived MVs promoted proliferation and migration of SCs in vitro and in vivo, which provided a therapeutic strategy for nerve regeneration. - Highlights: • M2 macrophages-derived MVs elevated migration and proliferation of SCs. • M2 macrophages-derived MVs up-regulated NFG and Laminin expression of SCs. • MiR-223 expression was increased in M2 macrophages-derived MVs. • MiR-223 inhibitor reduced migration and proliferation of SCs co-incubated with MVs. • MiR-223 inhibitor down-regulated NFG and Laminin levels of SCs co-incubated with MVs.« less

Towards a representation of priming on soil carbon decomposition in the global land biosphere model ORCHIDEE (version 1.9.5.2)

NASA Astrophysics Data System (ADS)

Guenet, Bertrand; Esteban Moyano, Fernando; Peylin, Philippe; Ciais, Philippe; Janssens, Ivan A.

2016-03-01

Priming of soil carbon decomposition encompasses different processes through which the decomposition of native (already present) soil organic matter is amplified through the addition of new organic matter, with new inputs typically being more labile than the native soil organic matter. Evidence for priming comes from laboratory and field experiments, but to date there is no estimate of its impact at global scale and under the current anthropogenic perturbation of the carbon cycle. Current soil carbon decomposition models do not include priming mechanisms, thereby introducing uncertainty when extrapolating short-term local observations to ecosystem and regional to global scale. In this study we present a simple conceptual model of decomposition priming, called PRIM, able to reproduce laboratory (incubation) and field (litter manipulation) priming experiments. Parameters for this model were first optimized against data from 20 soil incubation experiments using a Bayesian framework. The optimized parameter values were evaluated against another set of soil incubation data independent from the ones used for calibration and the PRIM model reproduced the soil incubations data better than the original, CENTURY-type soil decomposition model, whose decomposition equations are based only on first-order kinetics. We then compared the PRIM model and the standard first-order decay model incorporated into the global land biosphere model ORCHIDEE (Organising Carbon and Hydrology In Dynamic Ecosystems). A test of both models was performed at ecosystem scale using litter manipulation experiments from five sites. Although both versions were equally able to reproduce observed decay rates of litter, only ORCHIDEE-PRIM could simulate the observed priming (R2 = 0.54) in cases where litter was added or removed. This result suggests that a conceptually simple and numerically tractable representation of priming adapted to global models is able to capture the sign and magnitude of the priming of litter and soil organic matter.

Towards a representation of priming on soil carbon decomposition in the global land biosphere model ORCHIDEE (version 1.9.5.2)

NASA Astrophysics Data System (ADS)

Guenet, B.; Moyano, F. E.; Peylin, P.; Ciais, P.; Janssens, I. A.

2015-10-01

Priming of soil carbon decomposition encompasses different processes through which the decomposition of native (already present) soil organic matter is amplified through the addition of new organic matter, with new inputs typically being more labile than the native soil organic matter. Evidence for priming comes from laboratory and field experiments, but to date there is no estimate of its impact at global scale and under the current anthropogenic perturbation of the carbon cycle. Current soil carbon decomposition models do not include priming mechanisms, thereby introducing uncertainty when extrapolating short-term local observations to ecosystem and regional to global scale. In this study we present a simple conceptual model of decomposition priming, called PRIM, able to reproduce laboratory (incubation) and field (litter manipulation) priming experiments. Parameters for this model were first optimized against data from 20 soil incubation experiments using a Bayesian framework. The optimized parameter values were evaluated against another set of soil incubation data independent from the ones used for calibration and the PRIM model reproduced the soil incubations data better than the original, CENTURY-type soil decomposition model, whose decomposition equations are based only on first order kinetics. We then compared the PRIM model and the standard first order decay model incorporated into the global land biosphere model ORCHIDEE. A test of both models was performed at ecosystem scale using litter manipulation experiments from 5 sites. Although both versions were equally able to reproduce observed decay rates of litter, only ORCHIDEE-PRIM could simulate the observed priming (R2 = 0.54) in cases where litter was added or removed. This result suggests that a conceptually simple and numerically tractable representation of priming adapted to global models is able to capture the sign and magnitude of the priming of litter and soil organic matter.

Effect of Temperature on Biochemical Composition, Growth and Reproduction of the Ornamental Red Cherry Shrimp Neocaridina heteropoda heteropoda (Decapoda, Caridea)

PubMed Central

Tropea, Carolina; Stumpf, Liane; López Greco, Laura Susana

2015-01-01

The effect of water temperature on biochemical composition, growth and reproduction of the ornamental shrimp, Neocaridina heteropoda heteropoda, was investigated to determine the optimum temperature for its culture. The effect of embryo incubation temperature on the subsequent performance of juveniles was also evaluated. Ovigerous females and recently hatched juveniles (JI) were maintained during egg incubation and for a 90-day period, respectively, at three temperatures (24, 28 and 32°C). Incubation period increased with decreasing water temperature, but the number and size of JI were similar among treatments. At day 30 of the 90-day period, body weight and growth increment (GI) at 24°C were lower than those at 28 and 32°C. On subsequent days, GI at 24°C exceeded that at 28 and 32°C, leading to a similar body weight among treatments. These results suggest growth was delayed at 24°C, but only for 30 days after hatching. The lipid concentration tended to be lowest, intermediate and highest at 28, 32 and 24°C, respectively, possibly as a consequence of the metabolic processes involved in growth and ovarian maturation. Protein and glycogen concentrations were similar among treatments. Both the growth trajectory and biochemical composition of shrimps were affected by the temperature experienced during the 90-day growth period independently of the embryo incubation temperature. During the growth period, shrimps reached sexual maturity and mated, with the highest proportion of ovigerous females occurring at 28°C. All the females that matured and mated at 32°C lost their eggs, indicating a potentially stressful effect of high temperature on ovarian maturation. Based on high survival and good growth performance of shrimps at the three temperatures tested over the 90-day period it is concluded that N. heteropoda heteropoda is tolerant to a wide range of water temperatures, with 28°C being the optimum temperature for its culture. PMID:25768918

Non-Instrumented Incubation of a Recombinase Polymerase Amplification Assay for the Rapid and Sensitive Detection of Proviral HIV-1 DNA

PubMed Central

Lillis, Lorraine; Lehman, Dara; Singhal, Mitra C.; Cantera, Jason; Singleton, Jered; Labarre, Paul; Toyama, Anthony; Piepenburg, Olaf; Parker, Mathew; Wood, Robert; Overbaugh, Julie; Boyle, David S.

2014-01-01

Sensitive diagnostic tests for infectious diseases often employ nucleic acid amplification technologies (NAATs). However, most NAAT assays, including many isothermal amplification methods, require power-dependent instrumentation for incubation. For use in low resource settings (LRS), diagnostics that do not require consistent electricity supply would be ideal. Recombinase polymerase amplification (RPA) is an isothermal amplification technology that has been shown to typically work at temperatures ranging from 25–43°C, and does not require a stringent incubation temperature for optimal performance. Here we evaluate the ability to incubate an HIV-1 RPA assay, intended for use as an infant HIV diagnostic in LRS, at ambient temperatures or with a simple non-instrumented heat source. To determine the range of expected ambient temperatures in settings where an HIV-1 infant diagnostic would be of most use, a dataset of the seasonal range of daily temperatures in sub Saharan Africa was analyzed and revealed ambient temperatures as low as 10°C and rarely above 43°C. All 24 of 24 (100%) HIV-1 RPA reactions amplified when incubated for 20 minutes between 31°C and 43°C. The amplification from the HIV-1 RPA assay under investigation at temperatures was less consistent below 30°C. Thus, we developed a chemical heater to incubate HIV-1 RPA assays when ambient temperatures are between 10°C and 30°C. All 12/12 (100%) reactions amplified with chemical heat incubation from ambient temperatures of 15°C, 20°C, 25°C and 30°C. We also observed that incubation at 30 minutes improved assay performance at lower temperatures where detection was sporadic using 20 minutes incubation. We have demonstrated that incubation of the RPA HIV-1 assay via ambient temperatures or using chemical heaters yields similar results to using electrically powered devices. We propose that this RPA HIV-1 assay may not need dedicated equipment to be a highly sensitive tool to diagnose infant HIV-1 in LRS. PMID:25264766

Non-instrumented incubation of a recombinase polymerase amplification assay for the rapid and sensitive detection of proviral HIV-1 DNA.

PubMed

Lillis, Lorraine; Lehman, Dara; Singhal, Mitra C; Cantera, Jason; Singleton, Jered; Labarre, Paul; Toyama, Anthony; Piepenburg, Olaf; Parker, Mathew; Wood, Robert; Overbaugh, Julie; Boyle, David S

2014-01-01

Sensitive diagnostic tests for infectious diseases often employ nucleic acid amplification technologies (NAATs). However, most NAAT assays, including many isothermal amplification methods, require power-dependent instrumentation for incubation. For use in low resource settings (LRS), diagnostics that do not require consistent electricity supply would be ideal. Recombinase polymerase amplification (RPA) is an isothermal amplification technology that has been shown to typically work at temperatures ranging from 25-43°C, and does not require a stringent incubation temperature for optimal performance. Here we evaluate the ability to incubate an HIV-1 RPA assay, intended for use as an infant HIV diagnostic in LRS, at ambient temperatures or with a simple non-instrumented heat source. To determine the range of expected ambient temperatures in settings where an HIV-1 infant diagnostic would be of most use, a dataset of the seasonal range of daily temperatures in sub Saharan Africa was analyzed and revealed ambient temperatures as low as 10°C and rarely above 43°C. All 24 of 24 (100%) HIV-1 RPA reactions amplified when incubated for 20 minutes between 31°C and 43°C. The amplification from the HIV-1 RPA assay under investigation at temperatures was less consistent below 30°C. Thus, we developed a chemical heater to incubate HIV-1 RPA assays when ambient temperatures are between 10°C and 30°C. All 12/12 (100%) reactions amplified with chemical heat incubation from ambient temperatures of 15°C, 20°C, 25°C and 30°C. We also observed that incubation at 30 minutes improved assay performance at lower temperatures where detection was sporadic using 20 minutes incubation. We have demonstrated that incubation of the RPA HIV-1 assay via ambient temperatures or using chemical heaters yields similar results to using electrically powered devices. We propose that this RPA HIV-1 assay may not need dedicated equipment to be a highly sensitive tool to diagnose infant HIV-1 in LRS.

Cancer cell uptake behavior of Au nanoring and its localized surface plasmon resonance induced cell inactivation

NASA Astrophysics Data System (ADS)

Chu, Che-Kuan; Tu, Yi-Chou; Chang, Yu-Wei; Chu, Chih-Ken; Chen, Shih-Yang; Chi, Ting-Ta; Kiang, Yean-Woei; Yang, Chih-Chung

2015-02-01

Au nanorings (NRIs), which have the localized surface plasmon resonance (LSPR) wavelength around 1058 nm, either with or without linked antibodies, are applied to SAS oral cancer cells for cell inactivation through the LSPR-induced photothermal effect when they are illuminated by a laser of 1065 nm in wavelength. Different incubation times of cells with Au NRIs are considered for observing the variations of cell uptake efficiency of Au NRI and the threshold laser intensity for cell inactivation. In each case of incubation time, the cell sample is washed for evaluating the total Au NRI number per cell adsorbed and internalized by the cells based on inductively coupled plasma mass spectrometry measurement. Also, the Au NRIs remaining on cell membrane are etched with KI/I2 solution to evaluate the internalized Au NRI number per cell. The threshold laser intensities for cell inactivation before washout, after washout, and after KI/I2 etching are calibrated from the circular area sizes of inactivated cells around the illuminated laser spot center with various laser power levels. By using Au NRIs with antibodies, the internalized Au NRI number per cell increases monotonically with incubation time up to 24 h. However, the number of Au NRI remaining on cell membrane reaches a maximum at 12 h in incubation time. The cell uptake behavior of an Au NRI without antibodies is similar to that with antibodies except that the uptake NRI number is significantly smaller and the incubation time for the maximum NRI number remaining on cell membrane is delayed to 20 h. By comparing the threshold laser intensities before and after KI/I2 etching, it is found that the Au NRIs remaining on cell membrane cause more effective cancer cell inactivation, when compared with the internalized Au NRIs.

The effects of increased constant incubation temperature and cumulative acute heat shock exposures on morphology and survival of Lake Whitefish (Coregonus clupeaformis) embryos.

PubMed

Lee, Abigail H; Eme, John; Mueller, Casey A; Manzon, Richard G; Somers, Christopher M; Boreham, Douglas R; Wilson, Joanna Y

2016-04-01

Increasing incubation temperatures, caused by global climate change or thermal effluent from industrial processes, may influence embryonic development of fish. This study investigates the cumulative effects of increased incubation temperature and repeated heat shocks on developing Lake Whitefish (Coregonus clupeaformis) embryos. We studied the effects of three constant incubation temperatures (2°C, 5°C or 8°C water) and weekly, 1-h heat shocks (+3°C) on hatching time, survival and morphology of embryos, as these endpoints may be particularly susceptible to temperature changes. The constant temperatures represent the predicted magnitude of elevated water temperatures from climate change and industrial thermal plumes. Time to the pre-hatch stage decreased as constant incubation temperature increased (148d at 2°C, 92d at 5°C, 50d at 8°C), but weekly heat shocks did not affect time to hatch. Mean survival rates and embryo morphometrics were compared at specific developmental time-points (blastopore, eyed, fin flutter and pre-hatch) across all treatments. Constant incubation temperatures or +3°C heat-shock exposures did not significantly alter cumulative survival percentage (~50% cumulative survival to pre-hatch stage). Constant warm incubation temperatures did result in differences in morphology in pre-hatch stage embryos. 8°C and 5°C embryos were significantly smaller and had larger yolks than 2°C embryos, but heat-shocked embryos did not differ from their respective constant temperature treatment groups. Elevated incubation temperatures may adversely alter Lake Whitefish embryo size at hatch, but weekly 1-h heat shocks did not affect size or survival at hatch. These results suggest that intermittent bouts of warm water effluent (e.g., variable industrial emissions) are less likely to negatively affect Lake Whitefish embryonic development than warmer constant incubation temperatures that may occur due to climate change. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effect of ELISA kit manufacturing process and incubation time on progesterone concentration measured in dog serum for ovulation diagnosis - Short communication.

PubMed

Thuróczy, Julianna; Reiczigel, Jenő; Balogh, Lajos

2016-09-01

Twenty-two serum samples of healthy bitches were tested with the frozen and lyophilised version of the same ELISA kit (Quanticheck, Faculty of Veterinary Science, Budapest, Hungary). Samples were chosen on the basis of their progesterone (P4) concentrations, which were between 1.00 and 20.00 ng/mL. As it is well known, this range has the highest clinical relevance in ovulation diagnosis. Both types of microplates were read at 15-min intervals from the 15th until the 90th minute (min) of incubation, and the results were compared with those of frozen plates at 60 min of incubation as 100 percent. Lyophilised microplates gave on average 18 percent higher results than the frozen version at equal incubation times. The highest difference between lyophilised and frozen samples was observed at 45 and 60 min of incubation. Ninety-four percent of the reaction in the frozen microplate occurred in the first 15 min, and during the subsequent 30 min the reaction seemingly stopped. After the 45th min of incubation, this 94 percent increased to 108 percent in the subsequent 30 min, which remained the final approximate result at the end of the 90 min of incubation. In contrast to the frozen microplate, the measured concentration increased continuously in the lyophilised version and reached the highest level at the 60th min. The results of the lyophilised microplate reached the same level at 30 min of incubation as those of the frozen version at 60 min. In conclusion, a mechanical increase or decrease of the incubation time does not generate a linear change in the test results. This study demonstrated that the results of a series of samples collected from the same bitch cannot be compared if they are measured with different laboratory methods or different ELISA kits.

Inclusion of seminal plasma in sperm cryopreservation of Iberian pig.

PubMed

Gómez-Fernández, José; Gómez-Izquierdo, Emilio; Tomás, Cristina; González-Bulnes, Antonio; Sánchez-Sánchez, Raúl; de Mercado, Eduardo

2012-01-01

The aim of the present study was to evaluate the inclusion of seminal plasma (SP) in the freezing extender, trying to preserve as much as possible of SP with spermatozoa from Iberian pigs, thus improving the conservation of animal genetic resources of this breed. Experiment 1, evaluated the effect of substituting water with SP as diluent in the freezing media in different proportions (0%, 10%, 25%, 50%, 75% and 100%), over pre-freezing (at 10°C and 5°C) and post-thawing sperm quality. The results showed that over 50% of SP in the extender, significantly decreased sperm quality in comparison to the control sample (0% SP) and the samples with 10% and 25% of SP (P<0.05). No significant differences were found between the control sample and the samples with 10% and 25% SP (P>0.05), but treatment with 25% did not show significant differences between the time of incubation at 37°C after thawing (P>0.05), showing greater sperm quality resistance over time. Experiment 2, evaluated the effect of prolonged incubation period, until 480min (simulating the lifespan of sperm in the female genital tract), of sperm samples with 0%, 10% and 25% of SP. Treatment with 25% of SP maintained better sperm quality over time, compared to control sample. Significant differences were observed especially in the parameters of motility analysis (TMS, total motile spermatozoa; PMS: progressive motility spermatozoa. P<0.05). In Experiment 3, the effect of the presence of SP was evaluated during the thawing process. Although some differences were observed between treatments, these differences were not as clear as the previous experiments. In conclusion, replacement of 25% of the water by SP as diluent in the freezing extender could be considered the maximum percentage of inclusion, without harmful effects to the sperm. In addition, this proportion of SP maintained Iberian sperm quality for longer time when it was present during the freezing and thawing process. Copyright © 2012 Elsevier B.V. All rights reserved.

In vitro motility evaluation of aggregated cancer cells by means of automatic image processing.

PubMed

De Hauwer, C; Darro, F; Camby, I; Kiss, R; Van Ham, P; Decaesteker, C

1999-05-01

Set up of an automatic image processing based method that enables the motility of in vitro aggregated cells to be evaluated for a number of hours. Our biological model included the PC-3 human prostate cancer cell line growing as a monolayer on the bottom of Falcon plastic dishes containing conventional culture media. Our equipment consisted of an incubator, an inverted phase contrast microscope, a Charge Coupled Device (CCD) video camera, and a computer equipped with an image processing software developed in our laboratory. This computer-assisted microscope analysis of aggregated cells enables global cluster motility to be evaluated. This analysis also enables the trajectory of each cell to be isolated and parametrized within a given cluster or, indeed, the trajectories of individual cells outside a cluster. The results show that motility inside a PC-3 cluster is not restricted to slight motion due to cluster expansion, but rather consists of a marked cell movement within the cluster. The proposed equipment enables in vitro aggregated cell motility to be studied. This method can, therefore, be used in pharmacological studies in order to select anti-motility related compounds. The compounds selected by the equipment described could then be tested in vivo as potential anti-metastatic.

Effect of incubation temperature on the self-assembly of regenerated silk fibroin: a study using AFM.

PubMed

Zhong, Jian; Liu, Xunwei; Wei, Daixu; Yan, Juan; Wang, Ping; Sun, Gang; He, Dannong

2015-05-01

Understanding effect of temperature on the molecular self-assembly process will be helpful to unravel the structure-function relationship of biomolecule and to provide important information for the bottom-up approach to nanotechnology. In this work, the effect of incubation temperature on the secondary structures and morphological structures of regenerated silk fibroin (RSF) was systematically studied using atomic force microscopy and Fourier Transform infrared spectroscopy. The effect of incubation temperature on RSF self-assembly was dependent on RSF concentration. For the RSF solution with relatively low concentrations (15 μg/mL and 60 μg/mL), the increase of the incubation temperature mainly accelerated the formation and aggregation of antiparallel β-sheet protofibrils and decreased the formation of random coil protofilaments/globule-like molecules. For the RSF solution with relatively high concentrations (300 μg/mL and 1.5mg/mL), the increase of the incubation temperature mainly accelerated the formation and aggregation of antiparallel β-sheet RSF features (protofibrils and globule-like features) and decreased the formation of random coil bead-like features. This work implies that the morphology and conformation of biomacromolecules could be tuned by controlling the incubation temperature. Further, it will be beneficial to basic understanding of the nanoscale structure formation in different silk-based biomaterials. Copyright © 2015 Elsevier B.V. All rights reserved.

Effect of Incubation Time and Sucrose Addition on the Characteristics of Cheese Whey Yoghurt

NASA Astrophysics Data System (ADS)

Nurhartadi, E.; Utami, R.; Nursiwi, A.; Sari, A. M.; Widowati, E.; Sanjaya, A. P.; Esnadewi, E. A.

2017-04-01

The effect of incubation time and concentration of sucrose addition on the characteristics of cheese whey yogurt (lactic acid content, pH, total lactic acid bacteria, antioxidant activity, viscosity) and sensory characteristics (color, odor, flavor, consistency, and overalls) were investigated. The cheese whey yogurt fermentation process was carried out for 24h and 36h with the addition of sucrose 8, 10, and 12% (w/w) of total solid, respectively. The results showed that the lactic acid content, total lactic acid bacteria, antioxidant activity, and viscosity of cheese whey yogurt were affected by the incubation time and sucrose addition. The level of pH of yogurt which was incubated at 24h and 36h were relatively in the same levels, which were 4.51 up to 4.63. Due the sensory characteristic of cheese whey yogurt the panellists gave the high score for the cheese whey yogurt which was incubated at 24h and sucrose addition 12% (w/w) of total solid. The cheese whey yogurt has 0.41% lactic acid content; pH 4.51; 7.09 log total lactic acid bacteria cells / ml; 5.78% antioxidant activity; and 5.97 cP viscosity. The best sensory and physico-chemical characteristic of cheese whey yogurt was achieved by 24h incubation time and 12% concentration of sucrose addition.

Efficacy of ultra short sub-30 minute incubation of 5-aminolevulinic acid photodynamic therapy in vitro.

PubMed

Koo, Eugene; Austin, Evan; Mamalis, Andrew; Jagdeo, Jared

2017-08-01

The estimated incidence of cutaneous squamous cell carcinoma (SCC) is 700,000 cases per year. In the US, SCC incidence is highest among fair skinned adults older than 50 years of age. Thus, as the population ages, the reported number of SCCs will likely increase in the future. Photodynamic therapy (PDT) is an FDA approved therapy for treatment of actinic keratoses (AKs), a precursor to cutaneous SCC lesions. The FDA approved incubation time of the photosensitizing agent 5-aminolevulinic acid (ALA) is 14-18 hours. Recent studies have investigated short ALA incubation times of 1-3 hours with ALA and PDT demonstrating treatment success. Therefore, the question exists whether ALA incubation periods of less than 30 minutes are efficacious. Herein, we evaluate the efficacy of short ALA incubation periods by measuring apoptosis after 10, 15, and 20 minutes of ALA incubation. AG13145 normal human dermal fibroblasts HDFs were incubated with 10, 15, or 20 minute of ALA at various concentrations (0, 0.05, 0.075, 0.1, 0.25, 0.375, 0.5, 1, and 2 mM). After ALA incubation, samples were treated with 1,000 seconds (16 minutes 40 seconds) of Blu-U fluorescent blue light (417 ± 5 nm) for a fluence of 10 J/cm 2 . Immediately following treatment with blue light, samples were collected and stained for apoptosis and necrosis with annexin-V and 7-aminoactinomycin D (7-AAD), and then analyzed by flow cytometry. HDFs incubated with ALA for 10 minute at 36 °C followed by 10 J/cm 2 of blue light had no statistically significant changes in apoptosis. HDFs incubated with ALA for 15 or 20 minutes at 36 °C followed by 10 J/cm 2 of blue light had statistically significant increases in the percentages of cells positive for apoptosis in the 0.5, 1.0, and 2.0 mM ALA doses (P < 0.05). We found that incubation of ALA for at least 15 minutes followed by 10 J/cm 2 of blue light resulted in a statistically significant increase in apoptosis. Lasers Surg. Med. 49:592-598, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Modeling and investigation of submerged fermentation process to produce extracellular polysaccharide using Lactobacillus confusus.

PubMed

Thirugnanasambandham, K; Sivakumar, V; Prakash Maran, J

2014-12-19

The main objective of the present study is to investigate and optimize the Submerged fermentation (SMF) process parameters such as addition of coconut water, NaCl dose, incubation time and temperature on the production of extracellular polysaccharide (EPS) and biomass production using Lactobacillus confuses. Response surface methodology (RSM) coupled with four factors three level Box-Behnken design (BBD) was employed to model the SMF process. RSM analysis indicated good correspondence between experimental and predicted values. Three dimentional (3D) response surface plots were used to study the interactive effects of process variables on SMF process. The optimum process conditions for the maximum production of EPS and biomass were found to be as follows; addition of coconut water of 40%, NaCl dose of 15%, incubation time of 24h and temperature of 35°C. Under these conditions, 10.57 g/L of EPS and 3.9 g/L of biomass were produced. Copyright © 2014 Elsevier Ltd. All rights reserved.

A dark incubation period is important for Agrobacterium-mediated transformation of mature internode explants of sweet orange, grapefruit, citron, and a citrange rootstock.

PubMed

Marutani-Hert, Mizuri; Bowman, Kim D; McCollum, Greg T; Mirkov, T Erik; Evens, Terence J; Niedz, Randall P

2012-01-01

Citrus has an extended juvenile phase and trees can take 2-20 years to transition to the adult reproductive phase and produce fruit. For citrus variety development this substantially prolongs the time before adult traits, such as fruit yield and quality, can be evaluated. Methods to transform tissue from mature citrus trees would shorten the evaluation period via the direct production of adult phase transgenic citrus trees. Factors important for promoting shoot regeneration from internode explants from adult phase citrus trees were identified and included a dark incubation period and the use of the cytokinin zeatin riboside. Transgenic trees were produced from four citrus types including sweet orange, citron, grapefruit, and a trifoliate hybrid using the identified factors and factor settings. The critical importance of a dark incubation period for shoot regeneration was established. These results confirm previous reports on the feasibility of transforming mature tissue from sweet orange and are the first to document the transformation of mature tissue from grapefruit, citron, and a trifoliate hybrid.

Synthesis and Evaluation of Chlorinated Substrate Analogues for Farnesyl Diphosphate Synthase

PubMed Central

Heaps, Nicole A.; Poulter, C. Dale

2011-01-01

Substrate analogues for isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), where the C3 methyl groups were replaced by chlorine, were synthesized and evaluated as substrates for avian farnesyl diphosphate synthase (FPPase). The IPP analogue (3-ClIPP) was a co-substrate when incubated with dimethylallyl diphosphate (DMAPP) or geranyl diphosphate (GPP) to give the corresponding chlorinated analogues of geranyl diphosphate (3-ClGPP) and farnesyl diphosphate (3-ClFPP), respectively. No products were detected in incubations of 3-ClIPP with 3-ClDMAPP. Incubation of IPP with 3-ClDMAPP gave 11-ClFPP as the sole product. Values of KM3-ClIPP (with DMAPP) and KM3-ClDMAPP (with IPP) were similar to those for IPP and DMAPP, however values of kcat for both analogues were substantially lower. These results are consistent with a dissociative electrophilic alkylation mechanism where the rate-limiting step changes from heterolytic cleavage of the carbon-oxygen bond in the allylic substrate to alkylation of the double bond of the homoallylic substrate. PMID:21344952

Evaluation of Installation Time for SMASHmount by SMASHsolar

DOE Office of Scientific and Technical Information (OSTI.GOV)

The Department of Energy SunShot Incubator program provides early-stage assistance to help startup companies cross technological barriers to commercialization while encouraging private sector investment. The SunShot Incubator program aims to shorten the time it takes for a young business or company to develop an innovative product concept and make it commercially available, which includes product prototyping, deployment, and, potentially, manufacturing. SMASHsolar was selected as an Incubator awardee to develop a simple, snap-together, module-integrated photovoltaic (PV) mounting system in attempts to dramatically reduce the time, effort and skill needed to install rooftop solar. In support of this award, the National Renewablemore » Energy Laboratory worked with SMASHsolar to develop a procedure for evaluating the installation time required for the SMASHmount system vs. widely-available rail systems. Amongst several installations, NREL measured the following installation times, subject to the qualifications and conditions described later in this report. NREL found that the SMASHsolar SMASHmount system was installed between 15% and 37% faster than tested competing systems after one or two installations of the system.« less

A Dark Incubation Period Is Important for Agrobacterium-Mediated Transformation of Mature Internode Explants of Sweet Orange, Grapefruit, Citron, and a Citrange Rootstock

PubMed Central

Marutani-Hert, Mizuri; Bowman, Kim D.; McCollum, Greg T.; Mirkov, T. Erik; Evens, Terence J.; Niedz, Randall P.

2012-01-01

Background Citrus has an extended juvenile phase and trees can take 2–20 years to transition to the adult reproductive phase and produce fruit. For citrus variety development this substantially prolongs the time before adult traits, such as fruit yield and quality, can be evaluated. Methods to transform tissue from mature citrus trees would shorten the evaluation period via the direct production of adult phase transgenic citrus trees. Methodology/Principal Findings Factors important for promoting shoot regeneration from internode explants from adult phase citrus trees were identified and included a dark incubation period and the use of the cytokinin zeatin riboside. Transgenic trees were produced from four citrus types including sweet orange, citron, grapefruit, and a trifoliate hybrid using the identified factors and factor settings. Significance The critical importance of a dark incubation period for shoot regeneration was established. These results confirm previous reports on the feasibility of transforming mature tissue from sweet orange and are the first to document the transformation of mature tissue from grapefruit, citron, and a trifoliate hybrid. PMID:23082165

Transient Shifts of Incubation Temperature Reveal Immediate and Long-Term Transcriptional Response in Chicken Breast Muscle Underpinning Resilience and Phenotypic Plasticity.

PubMed

Naraballobh, Watcharapong; Trakooljul, Nares; Murani, Eduard; Brunner, Ronald; Krischek, Carsten; Janisch, Sabine; Wicke, Michael; Ponsuksili, Siriluck; Wimmers, Klaus

2016-01-01

Variations in egg incubation temperatures can have acute or long-term effects on gene transcription in avian species. Altered gene expression may, in turn, affect muscle traits in poultry and indirectly influence commercial production. To determine how changes in eggshell temperature affect gene expression, incubation temperatures were varied [36.8°C (low), 37.8°C (control), 38.8°C (high)] at specific time periods reflecting two stages of myogenesis [embryonic days (ED) 7-10 and 10-13]. Gene expression was compared between interventions and matching controls by microarrays in broiler breast muscle at ED10 or ED13 and post-hatch at day 35. Early (ED7-10) high incubation temperature (H10ΔC) resulted in 1370 differentially expressed genes (DEGs) in embryos. Ingenuity pathway analysis revealed temporary activation of cell maintenance, organismal development, and survival ability genes, but these effects were not maintained in adults. Late high incubation temperature (ED10-13) (H13ΔC) had slightly negative impacts on development of cellular components in embryos, but a cumulative effect was observed in adults, in which tissue development and nutrition metabolism were affected. Early low incubation temperature (L10ΔC) produced 368 DEGs, most of which were down-regulated and involved in differentiation and formation of muscle cells. In adults, this treatment down-regulated pathways of transcriptional processes, but up-regulated cell proliferation. Late low temperature incubation (L13ΔC) produced 795 DEGs in embryos, and activated organismal survival and post-transcriptional regulation pathways. In adults this treatment activated cellular and organ development, nutrition and small molecule activity, and survival rate, but deactivated size of body and muscle cells. Thermal interventions during incubation initiate immediate and delayed transcriptional responses that are specific for timing and direction of treatment. Interestingly, the transcriptional response to transiently decreased incubation temperature, which did not affect the phenotypes, prompts compensatory effects reflecting resilience. In contrast, higher incubation temperature triggers gene expression and has long-term effects on the phenotype. These mechanisms of considerable phenotypic plasticity contribute to the biodiversity and broaden the basis for managing poultry populations.

Thyroid Hormones Reduce Incubation Period without Developmental or Metabolic Costs in Murray River Short-Necked Turtles (Emydura macquarii).

PubMed

McGlashan, Jessica K; Thompson, Michael B; Van Dyke, James U; Spencer, Ricky-John

Metabolic processes are affected by both temperature and thyroid hormones in ectothermic vertebrates. Temperature is the major determinant of incubation length in oviparous vertebrates, but turtles can also alter developmental rate independent of temperature. Temperature gradients within natural nests cause different developmental rates of turtle embryos within nests. Despite temperature-induced reductions in developmental rate, cooler-incubated neonates often hatch synchronously with warmer siblings via metabolic compensation. The physiological mechanisms underlying metabolic compensation are unknown, but thyroid hormones may play a critical role. We applied excess triiodothyronine (T 3 ) to developing eggs of Murray River short-necked turtle (Emydura macquarii)-a species that exhibits metabolic compensation and synchronous hatching-to determine whether T 3 influences developmental rate and whether changes to incubation period incur metabolic costs. We measured heart rate, oxygen consumption and incubation period of eggs, and morphology and performance of hatchlings. Embryos that were exposed to T 3 pipped up to 3.5 d earlier than untreated controls, despite no change in total metabolic expenditure, and there were no treatment differences in hatchling morphology. Hatchlings treated with T 3 demonstrated similar righting ability to hatchlings from the control groups. Exposure to T 3 shortens incubation length by accelerating embryonic development but without statistically increasing embryonic metabolism. Thus, T 3 is a mechanism that cooler-incubated reptiles could use to accelerate their development to allow synchronous hatching with their warmer clutch mates but at little or no metabolic cost. Thus, metabolic compensation for synchronous hatching may not be metabolically expensive if T 3 is the underlying mechanism.

Temporal dynamics of fibrolytic and methanogenic rumen microorganisms during in situ incubation of switchgrass determined by 16S rRNA gene profiling.

PubMed

Piao, Hailan; Lachman, Medora; Malfatti, Stephanie; Sczyrba, Alexander; Knierim, Bernhard; Auer, Manfred; Tringe, Susannah G; Mackie, Roderick I; Yeoman, Carl J; Hess, Matthias

2014-01-01

The rumen microbial ecosystem is known for its biomass-degrading and methane-producing phenotype. Fermentation of recalcitrant plant material, comprised of a multitude of interwoven fibers, necessitates the synergistic activity of diverse microbial taxonomic groups that inhabit the anaerobic rumen ecosystem. Although interspecies hydrogen (H2) transfer, a process during which bacterially generated H2 is transferred to methanogenic Archaea, has obtained significant attention over the last decades, the temporal variation of the different taxa involved in in situ biomass-degradation, H2 transfer and the methanogenesis process remains to be established. Here we investigated the temporal succession of microbial taxa and its effect on fiber composition during rumen incubation using 16S rRNA amplicon sequencing. Switchgrass filled nylon bags were placed in the rumen of a cannulated cow and collected at nine time points for DNA extraction and 16S pyrotag profiling. The microbial community colonizing the air-dried and non-incubated (0 h) switchgrass was dominated by members of the Bacilli (recruiting 63% of the pyrotag reads). During in situ incubation of the switchgrass, two major shifts in the community composition were observed: Bacilli were replaced within 30 min by members belonging to the Bacteroidia and Clostridia, which recruited 34 and 25% of the 16S rRNA reads generated, respectively. A second significant shift was observed after 16 h of rumen incubation, when members of the Spirochaetes and Fibrobacteria classes became more abundant in the fiber-adherent community. During the first 30 min of rumen incubation ~13% of the switchgrass dry matter was degraded, whereas little biomass degradation appeared to have occurred between 30 min and 4 h after the switchgrass was placed in the rumen. Interestingly, methanogenic members of the Euryarchaeota (i.e., Methanobacteria) increased up to 3-fold during this period of reduced biomass-degradation, with peak abundance just before rates of dry matter degradation increased again. We hypothesize that during this period microbial-mediated fibrolysis was temporarily inhibited until H2 was metabolized into CH4 by methanogens. Collectively, our results demonstrate the importance of inter-species interactions for the biomass-degrading and methane-producing phenotype of the rumen microbiome-both microbially facilitated processes with global significance.

Evaluation of optimum roughage to concentrate ratio in maize stover based complete rations for efficient microbial biomass production using in vitro gas production technique.

PubMed

Reddy, Y Ramana; Kumari, N Nalini; Monika, T; Sridhar, K

2016-06-01

A study was undertaken to evaluate the optimum roughage to concentrate ratio in maize stover (MS) based complete diets for efficient microbial biomass production (EMBP) using in vitro gas production technique. MS based complete diets with roughage to concentrate ratio of 100:0, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, and 30:70 were formulated, and 200 mg of oven-dried sample was incubated in water bath at 39°C along with media (rumen liquor [RL] - buffer) in in vitro gas syringes to evaluate the gas production. The gas produced was recorded at 8 and 24 h of incubation. In vitro organic matter digestibility (IVOMD), metabolizable energy (ME), truly digestible organic matter (TDOM), partitioning factor (PF), and EMBP were calculated using appropriate formulae. Ammonia nitrogen and total volatile fatty acids (TVFAs) production were analyzed in RL fluid-media mixture after 24 h of incubation. In vitro gas production (ml) at 24 h incubation, IVOMD, ME, TDOM, TVFA concentration, and ammonia nitrogen production were increased (p<0.01) in proportion to the increase in the level of concentrate in the diet. Significantly (p<0.01) higher PF and EMBP was noticed in total mixed ration with roughage to concentrate ratio of 60:40 and 50:50 followed by 70:30 and 40:60. Based on the results, it was concluded that the MS can be included in complete rations for ruminants at the level of 50-60% for better microbial biomass synthesis which in turn influences the performance of growing sheep.

Use of Ancillary Carbapenemase Tests To Improve Specificity of Phenotypic Definitions for Carbapenemase-Producing Enterobacteriaceae.

PubMed

Miller, Shelley A; Hindler, Janet A; Chengcuenca, Angelo; Humphries, Romney M

2017-06-01

Carbapenemase-producing Enterobacteriaceae (CPE) are a significant threat to public health. In 2015, CDC revised the surveillance definition for CPE to include all Enterobacteriaceae resistant to any carbapenem tested. However, this definition is associated with poor specificity. We evaluated the performance of this definition, compared to carbapenemase PCR, for a collection of 125 Enterobacteriaceae We also investigated the impact of ancillary testing for carbapenemase of isolates that met the CDC CPE surveillance definition. The two ancillary tests evaluated were the Xpert Carba-R assay, a molecular test, and the carbapenem inactivation method (CIM). Two variables were evaluated for the CIM: suspension of organisms in double-distilled water (ddH 2 O) versus tryptic soy broth (TSB) to incubate disks, and incubation of plates for 6 h versus 18 to 20 h. The sensitivity and specificity of the Carba-R assay were 100% compared to the results of in-house PCR. The sensitivities of the CIM performed with TSB were 94.6% when read at 6 h and 97.7% when read at 18 to 20 h; the sensitivities with ddH 2 O were 88.0% when read at 6 h and 93.0% when incubated for 18 to 20 h. The specificity was 100% for all variables tested. Without ancillary testing, the sensitivity of the CDC definition was 98.9% for CPE, and the specificity was 6.1%. Testing isolates that screened positive by the CDC definition with the Xpert Carba-R did not change the sensitivity, and it improved the specificity to 100%. Similarly, the use of the CIM (TSB and 18 to 20 h of incubation) to confirm screen-positive isolates resulted in a sensitivity of 95.6% and specificity of 100%. Copyright © 2017 American Society for Microbiology.

Characterization of incubation experiments and development of an enrichment culture capable of ammonium oxidation under iron-reducing conditions

NASA Astrophysics Data System (ADS)

Huang, S.; Jaffé, P. R.

2015-02-01

Incubation experiments were conducted using soil samples from a forested riparian wetland where we have previously observed anaerobic ammonium oxidation coupled to iron reduction. Production of both nitrite and ferrous iron was measured repeatedly during incubations when the soil slurry was supplied with either ferrihydrite or goethite and ammonium chloride. Significant changes in the microbial community were observed after 180 days of incubation as well as in a continuous flow membrane reactor, using 16S rRNA gene PCR-denaturing gradient gel electrophoresis, 454 pyrosequencing, and real-time quantitative PCR analysis. We be Acidimicrobiaceae bacterium A6), belonging to the Acidimicrobiaceae family, whose closest cultivated relative is Ferrimicrobium acidiphilum (with 92% identity) and Acidimicrobium ferrooxidans (with 90% identity), might play a key role in this anaerobic biological process that uses ferric iron as an electron acceptor while oxidizing ammonium to nitrite. After ammonium was oxidized to nitrite, nitrogen loss proceeded via denitrification and/or anammox.

Undecompressed microbial populations from the deep sea.

PubMed Central

Jannasch, H J; Wirsen, C O; Taylor, C D

1976-01-01

Metabolic transformations of glutamate and Casamino Acids by natural microbial populations collected from deep waters (1,600 to 3,100 m) were studied in decompressed and undecompressed samples. Pressure-retaining sampling/incubation vessels and appropriate subsampling/incubation vessels and appropriate subsampling techniques permitted time course experiments. In all cases the metabolic activity in undecompressed samples was lower than it was when incubated at 1 atm. Surface water controls showed a reduced activity upon compression. The processes involving substrate incorporation into cell material were more pressure sensitive than was respiration. The low utilization of substrates, previously found by in situ incubations for up to 12 months, was confirmed and demonstrated to consist of an initial phase of activity, in the range of 5 to 60 times lower than the controls, followed by a stationary phase of virtually no substrate utilization. No barophilic growth response (higher rates at elevated pressure than at 1 atm) was recorded; all populations observed exhibition various degrees of barotolerance. Images PMID:791117

Structural phase transitions in niobium oxide nanocrystals

NASA Astrophysics Data System (ADS)

Yuvakkumar, R.; Hong, Sun Ig

2015-09-01

Niobium oxide nanocrystals were successfully synthesized employing the green synthesis method. Phase formation, microstructure and compositional properties of 1, 4 and 7 days incubation treated samples after calcinations at 450 °C were examined using X-ray diffraction, Raman, photoluminescence (PL), infrared, X-ray photoelectron spectra and transmission electron microscopic characterizations. It was observed that phase formation of Nb2O5 nanocrystals was dependent upon the incubation period required to form stable metal oxides. The characteristic results clearly revealed that with increasing incubation and aging, the transformation of cubic, orthorhombic and monoclinic phases were observed. The uniform heating at room temperature (32 °C) and the ligation of niobium atoms due to higher phenolic constituents of utilized rambutan during aging processing plays a vital role in structural phase transitions in niobium oxide nanocrystals. The defects over a period of incubation and the intensities of the PL spectra changing over a period of aging were related to the amount of the defects induced by the phase transition.

Progress of the Photovoltaic Technology Incubator Project Towards an Enhanced U.S. Manufacturing Base: Preprint

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ullal, H.; Mitchell, R.; Keyes, B.

In this paper, we report on the major accomplishments of the U.S. Department of Energy's (DOE) Solar Energy Technologies Program (SETP) Photovoltaic (PV) Technology Incubator project. The Incubator project facilitates a company's transition from developing a solar cell or PV module prototype to pilot- and large-scale U.S. manufacturing. The project targets small businesses that have demonstrated proof-of-concept devices or processes in the laboratory. Their success supports U.S. Secretary of Energy Steven Chu's SunShot Initiative, which seeks to achieve PV technologies that are cost-competitive without subsidies at large scale with fossil-based energy sources by the end of this decade. The Incubatormore » Project has enhanced U.S. PV manufacturing capacity and created more than 1200 clean energy jobs, resulting in an increase in American economic competitiveness. The investment raised to date by these PV Incubator companies as a result of DOE's $ 59 million investment totals nearly $ 1.3 billion.« less

Progress of the PV Technology Incubator Project Towards an Enhanced U.S. Manufacturing Base

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ullal, H.; Mitchell, R.; Keyes, B.

In this paper, we report on the major accomplishments of the U.S. Department of Energy's (DOE) Solar Energy Technologies Program (SETP) Photovoltaic (PV) Technology Incubator project. The Incubator project facilitates a company's transition from developing a solar cell or PV module prototype to pilot- and large-scale U.S. manufacturing. The project targets small businesses that have demonstrated proof-of-concept devices or processes in the laboratory. Their success supports U.S. Secretary of Energy Steven Chu's SunShot Initiative, which seeks to achieve PV technologies that are cost-competitive without subsidies at large scale with fossil-based energy sources by the end of this decade. The Incubatormore » Project has enhanced U.S. PV manufacturing capacity and created more than 1200 clean energy jobs, resulting in an increase in American economic competitiveness. The investment raised to date by these PV Incubator companies as a result of DOE's $ 59 million investment total nearly $ 1.3 billion.« less

Analysis of testosterone fatty acid esters in the digestive gland of mussels by liquid chromatography-high resolution mass spectrometry.

PubMed

Guercia, Cesare; Cianciullo, Piergiorgio; Porte, Cinta

2017-07-01

Several studies have indicated that up to 70% of the total steroids detected in molluscs are in the esterified form and that pollutants, by modifying the esterification of steroids with fatty acids, might act as endocrine disrupters. However, despite the strong physiological significance of this process, there is almost no information on which fatty acids form the steroid esters and how this process is modulated. This study (a) investigates the formation of fatty acid esters of testosterone in digestive gland microsomal fractions of the mussel Mytilus galloprovincialis incubated with either palmitoly-CoA or CoA and ATP, and (b) assesses whether the endocrine disruptor tributyltin (TBT) interferes with the esterification of testosterone. Analysis of testosterone esters was performed by liquid chromatography-high resolution mass spectrometry (UPLC-HRMS). When microsomal fractions were incubated with testosterone and palmitoly-CoA, the formation of testosterone palmitate was detected. However, when microsomes were incubated with CoA and ATP, and no exogenous activated fatty acid was added, the synthesis of 16:0, 16:1, 20:5 and 22:6 testosterone esters was observed. The presence of 100µM TBT in the incubation mixture did not significantly alter the esterification of testosterone. These results evidence the conjugation of testosterone with the most abundant fatty acids in the digestive gland microsomal fraction of mussels. Copyright © 2017 Elsevier Inc. All rights reserved.

Dissolution Mechanism for High Melting Point Transition Elements in Aluminum Melt

NASA Astrophysics Data System (ADS)

Lee, Young E.; Houser, Stephen L.

When added cold in aluminum melt, the alloying process for compacts of transition metal elements such as Mn, Fe, Cr, Ni, Ti, Cu, and Zn takes a sequence of incubation, exothermic reactions to form intermetallic compounds, and dispersion of the alloying elements into aluminum melt. The experiments with Cr compacts show that the incubation period is affected by the content of ingredient Al and size of compacts and by size of Cr particles. Incubation period becomes longer as the content of ingredient aluminum in compact decreases, and this prolonged incubation period negatively impacts the dissolution of the alloying elements in aluminum. Once liquid aluminum forms at reaction sites, the exothermic reaction takes place quickly and significantly raises the temperature of the compacts. As the result of it, the compacts swell in volume with a sponge like structure. Such porous structure encourages the penetration of liquid aluminum from the melt. The compacts become weak mechanically, and the alloying elements are dispersed and entrained in aluminum melt as discrete and small sized units. When Cr compacts are deficient in aluminum, the unreacted Cr particles are encased by the intermetallic compounds in the dispersed particles. They are carried in the melt flow and continue the dissolution reaction in aluminum. The entire dissolution process of Cr compacts completes within 10 to 15 minutes with a full recovery when the aluminum content is 10 to 20% in compacts.

Mouse Spermatozoa Contain a Nuclease that Is Activated by Pretreatment with EGTA and Subsequent Calcium Incubation

PubMed Central

Boaz, Segal M.; Dominguez, Kenneth; Shaman, Jeffrey A.; Ward, W. Steven

2009-01-01

We demonstrated that mouse spermatozoa cleave their DNA into ~50 kb loop-sized fragments with topoisomerase IIB when treated with MnCl2 and CaCl2 in a process we term sperm chromatin fragmentation (SCF). SCF can be reversed by EDTA. A nuclease then further degrades the DNA in a process we term sperm DNA degradation (SDD). MnCl2 alone could elicit this activity, but CaCl2 had no effect. Here, we demonstrate the existence of a nuclease in the vas deferens that can be activated by EGTA to digest the sperm DNA by SDD. Spermatozoa were extracted with salt and dithiothreitol to remove protamines and then incubated with EGTA. Next, the EGTA was removed and divalent cations were added. We found that Mn+2, Ca+2, or Zn+2 could each activate SDD in spermatozoa but Mg+2 could not. When the reaction was slowed by incubation on ice, EGTA pretreatment followed by incubation in Ca+2 elicited the reversible fragmentation of sperm DNA evident in SCF. When the reactions were then incubated at 37°C they progressed to the more complete degradation of DNA by SDD. EDTA could also be used to activate the nuclease, but required a higher concentration than EGTA. This EGTA-activatable nuclease activity was found in each fraction of the vas deferens plasma: in the spermatozoa, in the surrounding fluid, and in the insoluble components in the fluid. These results suggest that this sperm nuclease is regulated by a mechanism that is sensitive to EGTA, possibly by removing inhibition of a calcium binding protein. PMID:17879959

Effect of prenatal temperature conditioning of laying hen embryos: Hatching, live performance and response to heat and cold stress during laying period.

PubMed

Kamanli, S; Durmuş, I; Yalçın, S; Yıldırım, U; Meral, Ö

2015-07-01

This study was designed to determine the effect of prenatal temperature conditioning on hatching and live performance of laying chickens, and response to heat and cold stress during laying period. A total of 3600 eggs obtained from ATAK-S brown parent stock were incubated at control (37.5°C, CONT-Inc), cyclic low (36.5°C/6h/d from 10 to 18d of incubation, LOW-Inc) or high (38.5°C/6h/d from 10-18d of incubation, HIGH-Inc) incubation temperatures. Hatched chicks per incubation temperature were reared under standard rearing conditions up to 26wk. From 27 to 30wk, hens from each incubation temperature were divided into 3 environmentally controlled rooms and reared at control (20±2°C, CONT-Room), low (12±2°C, COLDS) or high (32±2°C, HEATS) temperatures. Hatching performance, body weight, egg production, and plasma triiodothyronine (T3) and thyroxine (T4) levels and oxidant and antioxidant activities were evaluated. The highest hatchability was for LOW-Inc chicks while HIGH-Inc chick had similar hatchability to CONT-Inc. There was no effect of incubation temperatures on plasma MDA, GSH-Px, activities and T4 concentrations on day of hatch. LOW- Inc chicks had higher SOD activities and T3 concentrations compared to the other groups. Although chick weight was similar among incubation temperature groups, CONT-Inc chicks were heavier than those cyclic incubation temperature groups until 12wk of age. Incubation temperature had no effect on sexual maturity age and weight and egg production of laying hens. From 27 to 30wk, regardless of incubation temperature, HEATS hens lost weight from day 0 to 10, had the highest cloacal temperatures and lowest feed consumption and egg production while COLDS hens had the lowest cloacal temperatures. At day 5, T4 level was higher in LOW-Inc hens at COLDS but it was higher in HIGH-Inc hens at HEATS compared to CONT-Inc. These data may suggest a modification in thyroid activity of hens that were conditioned during the incubation period. Moreover under COLDS condition, SOD production of LOW-Inc hens was higher than those of CONT- and HIGH-Inc hens indicating an induction in antioxidant enzyme activity. Nonetheless, prenatal temperature conditioning of laying hen embryos had no advantage on laying performance of hens under temperature stress conditions. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effects of soil water content and grass recycling on N2O emission in an urban lawn under laboratory incubation study

NASA Astrophysics Data System (ADS)

Nataningtyas, Dilin Rahayu; Morita, Shunsuke; Hatano, Ryusuke

2017-12-01

In the context of global warming, the increase of N2O gas production from the agricultural area has gained enhancing concern due to N2O is a potent greenhouse gas and an ozone depleted substance. While adding clipping grass has been accepted to replace N-fertilizer input in urban law management its effect on soil gas emission still questionable. A laboratory incubation study had been conducted to evaluate the effect of soil water content and grass recycling on greenhouse gas emission from an urban lawn. The soil samples were taken from Yurigahara Park, Sapporo, Hokkaido Japan. The 17 days at 25°C incubation study was started after adjusting soil water content to 35% and 50% with and without adding the clipping grass on soil surfaces. Greenhouse gas emissions were higher with the addition of grass, however, for NO and N2O considerably higher in 35% than 50% water content. The denitrification process was responsible for the N2O increase in this action. Soil chemical and microbial properties, pH, WEOC, NO3--N, NH4+-N and microbial biomass nitrogen (MBN) as well as N-grass content were also measured to know their correlation with N2O emission. The fine-scale heterogeneity occurred in the soil has impact on the variability of soil chemical properties that influenced the N2O emission. In the other hand, the effect of grass recycling appeared to increased soil N-inorganic contents and stimulated the N-gaseous concentration.

Re-Introduction of Lower Columbia River Chum Salmon into Duncan Creek, 2003-2004 Annual Report.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hillson, Todd D.

2004-09-01

Currently, two methods of reintroduction are being simultaneously evaluated at Duncan Creek. Recolonization is occurring by introducing adult chum salmon from the Lower Gorge (LG) population into Duncan Creek and allowing them to naturally reproduce. The supplementation strategy required adults to be collected and artificially spawned, incubated, reared, and released at the mouth of Duncan Creek. All eggs from the artificial crossings at Washougal Hatchery were incubated and the fry reared to release size at the hatchery. The Duncan Creek chum salmon project was very successful in 2003-04, providing knowledge and experience that will improve program execution in future years.more » The gear used to collect adult brood stock was changed from tangle nets to beach seines. This increased efficiency and the speed at which adults could be processed in the field, and most likely reduced stress on the adults handled. Certain weaknesses exposed in past seasons still exist and new ones were exposed (e.g. inadequate incubation and rearing space at Washougal Hatchery for any large salvage operation and having to move the rearing troughs outside the raceway in 2004). Egg-to-fry survival rates of 64% and 58% showed that the channels are functioning at the upper end of what can be expected from them. Possibly the most important event this season was the ability to strontium mark and release all naturally-produced fry from the spawning channels. Channel and floodplain modifications reduced the likelihood that floods will damage the channels and negatively impact survival rates.« less

Neonatal incubators: a toxic sound environment for the preterm infant?*.

PubMed

Marik, Paul E; Fuller, Christopher; Levitov, Alexander; Moll, Elizabeth

2012-11-01

High sound pressure levels may be harmful to the maturing newborn. Current guidelines suggest that the sound pressure levels within a neonatal intensive care unit should not exceed 45 dB(A). It is likely that environmental noise as well as the noise generated by the incubator fan and respiratory equipment may contribute to the total sound pressure levels. Knowledge of the contribution of each component and source is important to develop effective strategies to reduce noise within the incubator. The objectives of this study were to determine the sound levels, sound spectra, and major sources of sound within a modern neonatal incubator (Giraffe Omnibed; GE Healthcare, Helsinki, Finland) using a sound simulation study to replicate the conditions of a preterm infant undergoing high-frequency jet ventilation (Life Pulse, Bunnell, UT). Using advanced sound data acquisition and signal processing equipment, we measured and analyzed the sound level at a dummy infant's ear and at the head level outside the enclosure. The sound data time histories were digitally acquired and processed using a digital Fast Fourier Transform algorithm to provide spectra of the sound and cumulative sound pressure levels (dBA). The simulation was done with the incubator cooling fan and ventilator switched on or off. In addition, tests were carried out with the enclosure sides closed and hood down and then with the enclosure sides open and the hood up to determine the importance of interior incubator reverberance on the interior sound levels With all the equipment off and the hood down, the sound pressure levels were 53 dB(A) inside the incubator. The sound pressure levels increased to 68 dB(A) with all equipment switched on (approximately 10 times louder than recommended). The sound intensity was 6.0 × 10(-8) watts/m(2); this sound level is roughly comparable with that generated by a kitchen exhaust fan on high. Turning the ventilator off reduced the overall sound pressure levels to 64 dB(A) and the sound pressure levels in the low-frequency band of 0 to 100 Hz were reduced by 10 dB(A). The incubator fan generated tones at 200, 400, and 600 Hz that raised the sound level by approximately 2 dB(A)-3 dB(A). Opening the enclosure (with all equipment turned on) reduced the sound levels above 50 Hz by reducing the revereberance within the enclosure. The sound levels, especially at low frequencies, within a modern incubator may reach levels that are likely to be harmful to the developing newborn. Much of the noise is at low frequencies and thus difficult to reduce by conventional means. Therefore, advanced forms of noise control are needed to address this issue.

Comparison of some aspects of the in situ and in vitro methods in evaluation of neutral detergent fiber digestion.

PubMed

Krizsan, S J; Jančík, F; Ramin, M; Huhtanen, P

2013-02-01

The objective of the present study was to compare digestion rates (kd) of NDF for different feeds estimated with the in situ method or derived from an automated gas in vitro system. A meta-analysis was conducted to evaluate how in situ derived kd of NDF related to in vivo digestibility of NDF. Furthermore, in vitro true digestibility of the feed samples incubated within filter bags or dispersed in the medium was compared, and kd for insoluble and soluble components of those feeds were estimated. Four different concentrates and 4 forages were used in this study. Two lactating Swedish Red cows fed a diet of 60% grass silage and 40% concentrate on DM basis were used for in situ incubations and for collection of rumen fluid. The feed samples were ground through a 2.0-mm screen before the in situ incubations and a 1.0-mm screen before the in vitro gas incubations. In situ nylon bags were introduced into the rumen for determination of kd of NDF. Additional kinetic data were produced from isolated NDF and intact samples subjected to in vitro incubations in which gas production was recorded for 72 h. Samples were weighed in the bottles or within filter bags (for fiber and in vitro studies) that were placed in the bottles. The interaction between feed and method was significant (P < 0.01); kd of NDF for grass hay tended (P = 0.06) to be less whereas kd of NDF for alfalfa, barley grain, canola meal, and dried sugar beet pulp were greater (P < 0.01) when estimated with the in situ method than from gas production recordings. The meta-analysis suggested that in situ derived kd of NDF were biased and underestimated in vivo digestibility of NDF. Digestion rates of the intact samples were lower for all feeds, except for the hay, when incubated within the bags compared with dispersed in the medium (P < 0.01). Less OM and NDF were digested for all feeds when incubated within bags than dispersed in the medium (P < 0.01). It is concluded from the in vitro study that microbial activity within the bags is less than in the medium. Significant interactions between method (in situ vs. in vitro) and feed suggest that one or both methods result in biased estimates of digestion kinetics.

Antibody incubation at 37°C improves fluorescent immunolabeling in free-floating thick tissue sections.

PubMed

Xiao, Xia; Feng, Ya-Ping; Du, Bin; Sun, Han-Ru; Ding, You-Quan; Qi, Jian-Guo

2017-03-01

Fluorescent immunolabeling and imaging in free-floating thick (50-60 μm) tissue sections is relatively simple in practice and enables design-based non-biased stereology, or 3-D reconstruction and analysis. This method is widely used for 3-D in situ quantitative biology in many areas of biological research. However, the labeling quality and efficiency of standard protocols for fluorescent immunolabeling of these tissue sections are not always satisfactory. Here, we systematically evaluate the effects of raising the conventional antibody incubation temperatures (4°C or 21°C) to mammalian body temperature (37°C) in these protocols. Our modification significantly enhances the quality (labeling sensitivity, specificity, and homogeneity) and efficiency (antibody concentration and antibody incubation duration) of fluorescent immunolabeling of free-floating thick tissue sections.

Acetoacetate promotes the formation of fluorescent advanced glycation end products (AGEs).

PubMed

Bohlooli, Mousa; Ghaffari-Moghaddam, Mansour; Khajeh, Mostafa; Aghashiri, Zohre; Sheibani, Nader; Moosavi-Movahedi, Ali Akbar

2016-12-01

Acetoacetate (AA) is an important ketone body, which produces reactive oxygen species (ROS). Advanced glycation end products (AGEs) are defined as final products of glycation process whose production is influenced by the levels of ROS. The accumulation of AGEs in the body contributes to pathogenesis of many diseases including complications of diabetes, and Alzheimer's and Parkinson's disease. Here, we evaluated the impact of AA on production of AGEs upon incubation of human serum albumin (HSA) with glucose. The effect of AA on the AGEs formation of HSA was studied under physiological conditions after incubation with glucose for 35 days. The physical techniques including circular dichroism (CD) and fluorescence spectroscopy were used to assess the impact of AA on formation and structural changes of glycated HSA (GHSA). Our results indicated that the secondary and tertiary structural changes of GHSA were increased in the presence of AA. The fluorescence intensity measurements of AGEs also showed an increase in AGEs formation. Acetoacetate has an activator effect in formation of AGEs through ROS production. The presence of AA may result in enhanced glycation in the presence of glucose and severity of complications associated with accumulation of AGEs.

The Leucine Incorporation Method Estimates Bacterial Growth Equally Well in Both Oxic and Anoxic Lake Waters

PubMed Central

Bastviken, David; Tranvik, Lars

2001-01-01

Bacterial biomass production is often estimated from incorporation of radioactively labeled leucine into protein, in both oxic and anoxic waters and sediments. However, the validity of the method in anoxic environments has so far not been tested. We compared the leucine incorporation of bacterial assemblages growing in oxic and anoxic waters from three lakes differing in nutrient and humic contents. The method was modified to avoid O2 contamination by performing the incubation in syringes. Isotope saturation levels in oxic and anoxic waters were determined, and leucine incorporation rates were compared to microscopically observed bacterial growth. Finally, we evaluated the effects of O2 contamination during incubation with leucine, as well as the potential effects of a headspace in the incubation vessel. Isotope saturation occurred at a leucine concentration of above about 50 nM in both oxic and anoxic waters from all three lakes. Leucine incorporation rates were linearly correlated to observed growth, and there was no significant difference between oxic and anoxic conditions. O2 contamination of anoxic water during 1-h incubations with leucine had no detectable impact on the incorporation rate, while a headspace in the incubation vessel caused leucine incorporation to increase in both anoxic and O2-contaminated samples. The results indicate that the leucine incorporation method relates equally to bacterial growth rates under oxic and anoxic conditions and that incubation should be performed without a headspace. PMID:11425702

Efficacy of iontophoresis-assisted ablative fractional laser photodynamic therapy with short incubation time for the treatment of actinic keratosis: 12-month follow-up results of a prospective, randomised, comparative trial.

PubMed

Choi, Seung-Hwan; Kim, Tae-Hoon; Song, Ki-Hoon

2017-06-01

Iontophoresis is a transdermal drug-delivery technique that enhances the transport of ionic species across membranes and may have significant benefit for the treatment of actinic keratosis (AK) by ablative fractional laser-primed photodynamic therapy (AFL-PDT). The aims of this study were to compare the efficacy, recurrence rate, cosmetic outcome and safety of iontophoresis-assisted AFL-PDT with 2h of incubation vs. those of conventional AFL-PDT with 2- and 3-h incubation in patients with facial and scalp AK. Patients were randomly assigned to iontophoresis-assisted AFL-PDT with a 2-h incubation time (group A) and conventional AFL-PDT with a 2-h (group B) and 3-h (group C) incubation time. All patients underwent AFL-PDT, and group A patients were assigned to treatment with iontophoresis after methyl-aminolevulinate (MAL) application. After 2 or 3h, MAL-applied lesions were irradiated using a red light. Patients were followed up at 1-week, 3 months and 12 months after treatment. Efficacy, cosmetic outcomes and adverse events were assessed. In total, 41 patients (160 AK lesions) completed the study and were evaluated. Efficacy was significantly higher in Group A (88.7%) than in Group B (73.2%); the efficacy of groups A and C (92.2%) at 3 months follow-up was comparable. The recurrence rates were not significantly different between the groups at 12 months (P=0.841). The three groups did not differ in terms of cosmetic outcomes and safety. Iontophoresis-assisted AFL-PDT showed higher efficacy than AFL-PDT with short incubation time. Iontophoresis may effectively reduce the incubation time in AFL-PDT. Copyright © 2017 Elsevier B.V. All rights reserved.

Expression of thyroid hormone regulator genes in the yolk sac membrane of the developing chicken embryo

PubMed Central

TOO, Hanny Cho; SHIBATA, Mitsuhiro; YAYOTA, Masato; DARRAS, Veerle M.; IWASAWA, Atsushi

2017-01-01

Thyroid hormones (THs) are essential for the correct development of nearly every structure in the body from the very early stages of development, yet the embryonic thyroid gland is not functional at these stages. To clarify the roles of the egg yolk as a source of THs, the TH content in the yolk and the expression of TH regulator genes in the yolk sac membrane were evaluated throughout the 21-day incubation period of chicken embryos. The yolk TH content (22.3 ng triiodothyronine and 654.7 ng thyroxine per total yolk on day 4 of incubation) decreased almost linearly along with development. Real-time PCR revealed gene expression of transthyretin, a principal TH distributor in the chicken, and of a TH-inactivating iodothyronine deiodinase (DIO3), until the second week of incubation when the embryonic pituitary-thyroid axis is generally thought to start functioning. The TH-activating deiodinase (DIO2) and transmembrane transporter of thyroxine (SLCO1C1) genes were expressed in the last week of incubation, which coincided with a marked increase of circulating thyroxine and a reduction in the yolk sac weight. DIO1, which can remove iodine from inactive THs, was expressed throughout the incubation period. It is assumed that the chicken yolk sac inactivates THs contained abundantly in the yolk and supplies the hormones to the developing embryo in appropriate concentrations until the second week of incubation, while THs may be activated in the yolk sac membrane in the last week of incubation. Additionally, the yolk sac could serve as a source of iodine for the embryo. PMID:28652559

Effect of simethicone on the accuracy of the rapid urease test.

PubMed

Ng, F H; Wong, S Y; Kng, C; Chow, S L; Lai, K C; Ng, W F

1998-10-01

Simethicone, which is a principal ingredient in the defoaming agent used during gastroscopy, can inhibit the growth of Helicobacter pylori (MIC, 64-128 mg/l). This prospective study was designed to evaluate the in vivo and in vitro effects of simethicone on the accuracy of the rapid urease test (RUT). In the in vivo study, three sets of gastric biopsies (two from the antrum, and one from the corpus) were taken from 75 patients. The first set was examined histologically, and the second set was used for the RUT (pre-simethicone RUT). Then, 25 ml simethicone (1200 mg/l) was introduced into the stomach for a contact time of 1 min. A third set of gastric biopsies was taken for the RUT (post-simethicone RUT). In the in vitro clinical study, 41 patients were recruited. The first set of gastric biopsies was used for the RUT (pre-incubation RUT). The second set was incubated in 1 ml of simethicone for 5 min before being used for the RUT (post-incubation RUT). In the spectrophotometric study, urease activity before and after incubation in simethicone for 5 min was quantified in 12 patients by measuring the absorbance at 560 nm. Reading at 15 min, the concordance rate between the pre-simethicone and post-simethicone RUT was 98%. In the in vitro clinical study, the concordance rate between the pre-incubation and post-incubation RUT was 97%. The spectophotometric study showed a significant reduction of 43% in urease activity after incubation in simethicone. The application of a defoaming agent containing simethicone does not affect the accuracy of the RUT. However, simethicone modestly suppresses urea hydrolysis.

MHC Class II Activation and Interferon-γ Mediate the Inhibition of Neutrophils and Eosinophils by Staphylococcal Enterotoxin Type A (SEA).

PubMed

Ferreira-Duarte, Ana P; Pinheiro-Torres, Anelize S; Anhê, Gabriel F; Condino-Neto, Antônio; Antunes, Edson; DeSouza, Ivani A

2017-01-01

Staphylococcal enterotoxins are classified as superantigens that act by linking T-cell receptor with MHC class II molecules, which are expressed on classical antigen-presenting cells (APC). Evidence shows that MHC class II is also expressed in neutrophils and eosinophils. This study aimed to investigate the role of MHC class II and IFN-γ on chemotactic and adhesion properties of neutrophils and eosinophils after incubation with SEA. Bone marrow (BM) cells obtained from BALB/c mice were resuspended in culture medium, and incubated with SEA (3-30 ng/ml; 1-4 h), after which chemotaxis and adhesion were evaluated. Incubation with SEA significantly reduced the chemotactic and adhesive responses in BM neutrophils activated with IL-8 (200 ng/ml). Likewise, SEA significantly reduced the chemotactic and adhesive responses of BM eosinophils activated with eotaxin (300 ng/ml). The inhibitory effects of SEA on cell chemotaxis and adhesion were fully prevented by prior incubation with an anti-MHC class II blocking antibody (2 μg/ml). SEA also significantly reduced the intracellular Ca 2+ levels in IL-8- and eotaxin-activated BM cells. No alterations of MAC-1, VLA4, and LFA-1α expressions were observed after SEA incubation. In addition, SEA elevated by 3.5-fold ( P < 0.05) the INF-γ levels in BM cells. Incubation of BM leukocytes with IFN-γ (10 ng/ml, 2 h) reduced both neutrophil and eosinophil chemotaxis and adhesion, which were prevented by prior incubation with anti-MHC class II antibody (2 μg/ml). In conclusion, SEA inhibits neutrophil and eosinophil by MHC class II-dependent mechanism, which may be modulated by concomitant release of IFN-γ.

Use of supplemental food by breeding Ross's Geese and Lesser Snow Geese: Evidence for variable anorexia

USGS Publications Warehouse

Gloutney, M.L.; Alisauskas, R.T.; Hobson, K.A.; Afton, A.D.

1999-01-01

Recent research suggests that foods eaten during laying and incubation play a greater role in supplying energy and nutrients to arctic-nesting geese than previously believed. We conducted food-supplementation experiments with Ross's Geese (Chen rossii) and Lesser Snow Geese (C. caerulescens) geese to evaluate: (1) if supplemental food was consumed by laying and incubating geese, (2) how food consumption influenced mass dynamics of somatic tissues of breeding geese, (3) if patterns of mass loss were consistent with fasting adaptations, and (4) whether energetic constraints would cause smaller Ross's Geese to consume more food relative to their body size than would larger Snow Geese. Quantity of supplemental food eaten by both species during laying and incubation was highly variable among individuals. Consumption of supplemental food during laying resulted in differences in overall body composition between control and treatment females. Treatment female Ross's Geese completed laying at a higher mass and with more abdominal fat than controls, whereas treatment female Snow Geese completed laying with heavier breast muscles and hearts. Overall body composition did not differ between control and treatment geese (both sexes and species) at the end of incubation, but treatment geese had heavier hearts than control geese. This suggests that treatment females did not rely to the same extent on metabolic adaptations associated with anorexia to meet energetic costs of incubation as did controls. Stable-nitrogen isotope analysis revealed patterns of protein maintenance during incubation consistent with metabolic adaptations to prolonged fasting. Our prediction that energetic constraints would cause smaller Ross's Geese to consume more food relative to their size than would Snow Geese was not supported. Mass-specific food consumption by Ross's Geese was 30% lower than that of Snow Geese during laying and 48% higher during incubation.

Greater absolute rates of N2O production and consumption with soil warming dwarf variations in denitrification enzyme temperature sensitivities across seasons

NASA Astrophysics Data System (ADS)

Tiemann, L. K.; Billings, S. A.

2010-12-01

Investigators appreciate the important role that nitrate (NO3-) and soil moisture availability can play in governing net N2O production from soils. However, a large knowledge gap remains surrounding the drivers of soil N2O consumption and the role of microbial adaptation to changing environmental conditions in governing both N2O production and consumption. Net N2O soil efflux can be correlated with temperature, but little is known about the influence of temperature on gross rates of N2O production vs. consumption. Further, we do not understand how microbial communities responsible for these processes adapt or acclimate to soil warming. To investigate whether temperature alters the denitrifier-mediated fate of NO3- lost via N2O or N2, and if any such effect changes across seasons, we incubated soil collected in three seasons at four temperatures with and without 15N-enriched nitrate for 26 hours. Incubations were conducted in an anaerobic environment flushed with helium to permit detection of N2O and N2, and those gases’ δ15N. Temperature positively influenced CO2 production resulting from anaerobic processes. Maximum values of net N2O production were positively influenced by incubation and seasonal temperature, and the maximum rate of net N2O production occurred relatively early at warmer incubation temperatures. We also observed greater N2O:N2 ratios early in the incubations at warmer incubation temperatures. Isotope data are consistent with these trends. For those soils receiving the 15N label, differences in δ15N2O between early and late in the incubations were increasingly negative, and differences in δ15N2 increasingly positive, as temperature increased. Q10 values for N2O production and consumption exhibited increasing similarities as seasons progressed, with June N2O production and consumption Q10 values being nearly identical. These data provide convincing evidence that: a) increasing temperatures can induce denitrifying communities to perform complete denitrification (i.e. consumption of gross N2O production into N2) to a greater degree, and permit release of a relatively smaller proportion of the nitrate they consumed as N2O; b) the suite of enzymes responsible for N2O production and the one enzyme responsible for its consumption exhibit differential temperature sensitivities in their production and expression during winter months, but the sensitivity of these processes converges during warmer seasons; c) in spite of the smaller proportion of NO3- released as N2O with warming, warming’s positive influence on the amount of NO3- transformed by denitrifying organisms resulted in far greater absolute quantities of N2O released with incubation and seasonal warming. Continuing work explores the influence that temperature may exert on the relative abundances of denitrifying populations and their gene expression, and links these microbial characteristics to denitrification processes with warming. These data signify the importance of understanding enzyme kinetics in concert with microbial adaptation and acclimation as a factor governing the net fluxes of N2O from soil vs. its transformation into N2 with warming.

Tunable Light-Guide Image Processing Snapshot Spectrometer (TuLIPSS) for Earth and Moon Observations

NASA Astrophysics Data System (ADS)

Tkaczyk, T. S.; Alexander, D.; Luvall, J. C.; Wang, Y.; Dwight, J. G.; Pawlowsk, M. E.; Howell, B.; Tatum, P. F.; Stoian, R.-I.; Cheng, S.; Daou, A.

2018-02-01

A tunable light-guide image processing snapshot spectrometer (TuLIPSS) for Earth science research and observation is being developed through a NASA instrument incubator project with Rice University and Marshall Space Flight Center.

Detection of Legionella pneumophila on clinical samples and susceptibility assessment by flow cytometry.

PubMed

Faria-Ramos, I; Costa-de-Oliveira, S; Barbosa, J; Cardoso, A; Santos-Antunes, J; Rodrigues, A G; Pina-Vaz, C

2012-12-01

Culture in selective media represents the standard diagnostic method to confirm Legionella pneumophila infection, despite requiring a prolonged incubation period; antigen detection by immunofluorescence (IFS) and molecular techniques are also available, but they do not allow antimicrobial susceptibility evaluation. Our objective was to optimise flow cytometry (FC) protocols for the detection of L. pneumophila in respiratory samples and for susceptibility evaluation to first-line drugs. In order to optimise the FC protocol, a specific monoclonal antibody, conjugated with fluorescein isothiocyanate (FITC), was incubated with type strain L. pneumophila ATCC 33152. The limit of detection was established by analysing serial dilutions of bacterial suspension; specificity was assayed using mixtures of prokaryotic and eukaryotic microorganisms. The optimised FC protocol was used to assess 50 respiratory samples and compared with IFS evaluation. The susceptibility profile to erythromycin, ciprofloxacin and levofloxacin was evaluated by FC using propidium iodide and SYBR Green fluorescent dyes; the results were compared with the Etest afterwards. The optimal specific antibody concentration was 20 μg/ml; 10(2)/ml Legionella organisms were detected by this protocol and no cross-reactions with other microorganisms were detected. The five positive respiratory samples (10 %) determined by IFS were also detected by FC, showing 100 % correlation. After 1 h of incubation at 37 °C with different antimicrobials, SYBR Green staining could discriminate between treated and non-treated cells. A novel flow cytometric approach for the detection of L. pneumophila from clinical samples and susceptibility evaluation is now available, representing an important step forward for the diagnosis of this very relevant agent.

Formation of HDL-like complexes from apolipoprotein A-I(M) and DMPC.

PubMed

Suurkuusk, M; Singh, S K

2000-01-20

Conditions for the preparation of reconstituted high density lipoproteins (HDLs) by incubation of the synthetic lipid dimyristoylphosphatidylcholine (DMPC) and recombinant apolipoprotein A-I(M) have been investigated as a function of ratio of incubation lipid to protein, incubation temperature and the lipid form (multilamellar (MLV) or small unilamellar (SUV) vesicles). The size distributions of the resultant lipid-protein complex particles from various incubations have been evaluated by native gel electrophoresis. Structural changes of the protein after incorporation into these complex particles have been estimated by CD. Thermal characteristics of the particles has been examined by DSC and correlated with CD results. Titration calorimetry has been used to obtain interaction parameters based on a simplified binding model. It is hypothesized that the major enthalpic step in the production of rHDLs is the primary association step between protein and lipid vesicles. It has been shown that by raising the temperature and incubation ratio, the formation of rHDL particles can be directed towards smaller size and a narrower size distribution. The results have been described on the basis of a model where formation of discoidal particles requires prior saturation of vesicle surface area by adsorbed protein, thus explaining differences between particles formed from MLVs and SUVs.

Revitalizing the HRD Function.

ERIC Educational Resources Information Center

Lawrie, John

1986-01-01

A proactive approach to human resource development requires assessment of skills and attitudes, review of the diagnosis, an incubation period, implementation of training, and evaluation of results. (SK)

Environmental Regulation of Microbial Community Structure

NASA Technical Reports Server (NTRS)

Bebout, Leslie; DesMarais, D.; Heyenga, G.; Nelson, F.; DeVincenzi, D. (Technical Monitor)

2002-01-01

Most naturally occurring microbes live in complex microbial communities consisting of thousands of phylotypes of microorganisms living in close proximity. Each of these draws nutrients from the environment and releases metabolic waste products, which may in turn serve as substrates for other microbial groups. Gross environmental changes, such as irradiance level, hydrodynamic flow regime, temperature or water chemistry can directly affect the productivity of some community members, which in turn will affect other dependent microbial populations and rate processes. As a first step towards the development of "standard" natural communities of microorganisms for a variety of potential NASA applications, we are measuring biogeochemical cycling in artificially structured communities of microorganisms, created using natural microbial mat communities as inoculum. The responses of these artificially assembled communities of microorganisms to controlled shifts in ecosystem incubation conditions is being determined. This research requires close linking of environmental monitoring, with community composition in a closed and controlled incubation setting. We are developing new incubation chamber designs to allow for this integrated approach to examine the interplay between environmental conditions, microbial community composition and biogeochemical processes.

Screening and optimization of some inorganic salts for the production of ergot alkaloids from Penicillium species using surface culture fermentation process.

PubMed

Shahid, Memuna Ghafoor; Nadeem, Muhammad; Baig, Shahjehan; Cheema, Tanzeem Akbar; Atta, Saira; Ghafoor, Gul Zareen

2016-03-01

The present study deals with the production of ergot alkaloids from Penicillium commune and Penicillium citrinum, using surface culture fermentation process. Impact of various inorganic salts was tested on the production of ergot alkaloids during the optimization studies of fermentation medium such as impact of various concentration levels of succinic acid, ammonium chloride, MgSO4, FeSO4, ZnSO4, pH and the effect of various incubation time periods was also determined on the production of ergot alkaloids from Penicillium commune and Penicillium citrinum. Highest yield of ergot alkaloids was obtained when Penicillium commune and Penicillium citrinum that were grown on optimum levels of ingredients such as 2 g succinic acid, 1.5 and 2 g NH4Cl, 1.5 g MgSO4, 1 g FeSO4, 1 and 1.5 g ZnSO4 after 21 days of incubation time period using pH 5 at 25(°)C incubation temperature in the fermentation medium. Ergot alkaloids were determined using Spectrophotometry and Thin Layer Chromatography (TLC) techniques.

Egg storage duration and hatch window affect gene expression of nutrient transporters and intestine morphological parameters of early hatched broiler chicks.

PubMed

Yalcin, S; Gursel, I; Bilgen, G; Izzetoglu, G T; Horuluoglu, B H; Gucluer, G

2016-05-01

In recent years, researchers have given emphasis on the differences in physiological parameters between early and late hatched chicks within a hatch window. Considering the importance of intestine development in newly hatched chicks, however, changes in gene expression of nutrient transporters in the jejunum of early hatched chicks within a hatch window have not been studied yet. This study was conducted to determine the effects of egg storage duration before incubation and hatch window on intestinal development and expression of PepT1 (H+-dependent peptide transporter) and SGLT1 (sodium-glucose co-transporter) genes in the jejunum of early hatched broiler chicks within a 30 h of hatch window. A total of 1218 eggs obtained from 38-week-old Ross 308 broiler breeder flocks were stored for 3 (ES3) or 14 days (ES14) and incubated at the same conditions. Eggs were checked between 475 and 480 h of incubation and 40 chicks from each egg storage duration were weighed; chick length and rectal temperature were measured. The chicks were sampled to evaluate morphological parameters and PepT1 and SGLT1 expression. The remaining chicks that hatched between 475 and 480 h were placed back in the incubator and the same measurements were conducted with those chicks at the end of hatch window at 510 h of incubation. Chick length, chick dry matter content, rectal temperature and weight of small intestine segments increased, whereas chick weight decreased during the hatch window. The increase in the jejunum length and villus width and area during the hatch window were higher for ES3 than ES14 chicks. PepT1 expression was higher for ES3 chicks compared with ES14. There was a 10.2 and 17.6-fold increase in PepT1 and SGLT1 expression of ES3 chicks at the end of hatch window, whereas it was only 2.3 and 3.3-fold, respectively, for ES14 chicks. These results suggested that egg storage duration affected development of early hatched chicks during 30 h of hatch window. It can be concluded that the ES14 chicks would be less efficiently adapted to absorption process for carbohydrates and protein than those from ES3 at the end of the hatch window.

ANAEROBIC SOIL DISINFESTATION IN MICROCOSMS OF TWO SANDY SOILS.

PubMed

Stremińska, M A; Runia, W T; Termorshuizen, A J; Feil, H; Van Der Wurff, A W G

2014-01-01

In recent years, anaerobic soil disinfestation (ASD) has been proposed as an alternative control method of soil-borne plant pathogens. It involves adding a labile carbon source, irrigating the soil to stimulate decomposition of organic material and then covering the soil with air-tight plastic to limit gas exchange. During the ASD process, soil microorganisms switch from aerobic to anaerobic metabolism. As a result, by-products of anaerobic metabolism are released into the soil environment such as various organic acids and gases. These by-products are reported to have a negative effect on survival of soil-borne plant pathogens. However, the efficacy of ASD to reduce soil-borne pathogens in practice may vary significantly. Therefore, we studied the efficacy of the ASD process in two different soils. In addition, it was investigated whether a pre-treatment with an anaerobic bacterial inoculum prior to ASD affected the efficacy of the process. Two sandy soils (dune sand and glacial sand) were inoculated in 2 L soil microcosms. We tested the efficacy of ASD treatment against the potato cyst nematode Globodera pallida. For each soil, three treatments were used: control treatment (no Herbie addition, aerobic incubation), ASD 1 (organic substrate addition, anaerobic incubation) and ASD 2 (organic substrate and anaerobic bacterial inoculum addition, anaerobic incubation). Soil microcosms were incubated in the dark at 20°C for two weeks. We observed that anaerobic soil disinfestation treatments were highly effective against Potato Cyst Nematode (PCN), with pathogen being eradicated totally in all but one ASD treatment (glacial sand ASD2) within two weeks. The relative abundance of Firmicutes (spore-forming bacteria, often fermentative) in total bacteria increased significantly in ASD treated soils. Numbers of these bacteria correlated positively with increased concentrations of acetic and butyric acids in soil water phase in ASD treatments.

Cytotoxic effects of resin-modified orthodontic band adhesives. Are they safe?

PubMed

Malkoc, Siddik; Corekci, Bayram; Botsali, Hayriye Esra; Yalçin, Muhammet; Sengun, Abdülkadir

2010-09-01

To evaluate the cytotoxic effects of three different resin-modified orthodontic band adhesives. Three resin-modified orthodontic band adhesives (Bisco Ortho Band Paste LC, Multi-Cure Glass Ionomer Band Cement, and Transbond Plus Light Cure Band Adhesive) were prepared and the samples were extracted in 3 mL of Basal Medium Eagle with 10% newborn calf serum for 24 hours. The L929 cells were plated (25,000 cells/mL) in wells of 96-well dishes and maintained in a humidified incubator for 24 hours at 37 degrees C, 5% CO(2), and 95% air. After 24-hour incubation of the cells, the incubation medium was replaced by the immersed medium in which the samples were stored. Then L929 cells were incubated in contact with eluates for 24 hours. The cell mitochondrial activity was evaluated by the methyltetrazolium test. Twelve wells were used for each specimen, and methyltetrazolium tests were applied two times. The data were statistically analyzed using one-way analysis of variance and Tukey Honestly Significantly Different tests. Results with L929 fibroblasts demonstrated that all freshly prepared resin-modified orthodontic band adhesive materials reduced vital cell numbers (P > .05), in comparison to the control group. Our data demonstrate that all materials showed significant cytotoxicity compared to the control group. The results indicate that all materials showed significant cytotoxicity compared to the control group, and further studies using different test methods are needed for all resin-modified orthodontic band adhesives.

Characterization of egg white antibacterial properties during the first half of incubation: A comparative study between embryonated and unfertilized eggs.

PubMed

Guyot, N; Réhault-Godbert, S; Slugocki, C; Harichaux, G; Labas, V; Helloin, E; Nys, Y

2016-12-01

Egg white is an important contributor to the protection of eggs against bacterial contaminations during the first half of incubation (day zero to 12), prior to the egg white transfer into the amniotic fluid to be orally absorbed by the embryo. This protective system relies on an arsenal of antimicrobial proteins and on intrinsic physicochemical properties that are generally unfavorable for bacterial multiplication and dissemination. Some changes in these parameters can be observed in egg white during egg storage and incubation. The aim of this work was to characterize changes in the antibacterial potential of egg white in embryonated eggs (FE) during the first half of incubation using unfertilized eggs (UF) as controls. Egg white samples were collected at day zero, 4, 8, and 12 and analyzed for pH, protein concentration, and protein profile. Antibacterial properties of egg white proteins were evaluated against Listeria monocytogenes, Streptococcus uberis, Staphylococcus aureus, Escherichia coli, and Salmonella Enteritidis. During incubation, differential variations of egg white pH and protein concentrations were observed between UF and FE. At equal protein concentrations, similar activities against L. monocytogenes and S. uberis were observed for FE and UF egg white proteins. A progressive decline in these activities, however, was observed over incubation time, regardless of the egg group (UF or FE). SDS-PAGE analysis of egg white proteins during incubation revealed discrete changes in the profile of major proteins, whereas the stability of some less abundant antimicrobial proteins seemed more affected. To conclude, the antibacterial activity of egg white proteins progressively decreased during the first half of egg incubation, possibly resulting from the alteration of specific antimicrobial proteins. This apparent decline may be partly counterbalanced in embryonated eggs by the increase in egg white protein concentration. The antibacterial potential of egg white is very effective during early stages of embryonic development but its alteration during incubation suggests that extra-embryonic structures could then progressively ensure protective functions. © 2016 Poultry Science Association Inc.

Effects of LED lighting during incubation on layer and broiler hatchability, chick quality, stress susceptibility and post-hatch growth.

PubMed

Huth, Jesse C; Archer, Gregory S

2015-12-01

Providing light during incubation has been shown to affect hatchability, but the use of LED lights has not been evaluated. This experiment evaluated the effects of LED lighting during embryogenesis on White Leghorn and commercial broiler eggs. To determine this, two experiments were conducted, the first using White Leghorn eggs (N=3456) and the second using commercial broiler eggs (N=3456) where eggs were incubated 12 h of light and 12 h of darkness (LED) or complete darkness (DARK); the light level was 250 lux. Hatchability, embryo mortality, and chick quality were measured in both studies, and a subset of one of the broiler egg trials were grown out to investigate fear and stress parameters. There was no effect (P>0.05) on hatchability of layer eggs; however, there was a difference (P=0.02) observed in chick quality, with the LED group having more chicks (75.34%) with no defects than the DARK group (56.53%). Broiler eggs exposed to LED light showed an increase in hatchability (90.12%, P=0.03) and an increase in no-defect chick percentage (86.12%, P=0.04) at hatch compared to the DARK chicks (85.76% and 69.43%, respectively). Differences were observed between treatments during the 14 d grow-out. The LED birds had lower (P<0.05) physical asymmetry (0.90±0.05 mm) and heterophil/lymphocyte ratios (0.279±0.021), indicating that they were less susceptible to stress than the DARK birds (1.16±0.07 mm and 0.347±0.021, respectively). There was no difference (P>0.05) observed between treatments in growth, FCR, or fear measures at 14 d. These results indicate that providing LED light during incubation can improve chick quality in both white layer and broiler eggs; however, it only appears to improve hatchability in broilers, which could be related to shell pigmentation. It was also demonstrated that providing LED light during incubation can reduce the stress susceptibility of broilers post-hatch. Utilizing light during incubation may be useful tool for the poultry industry. © 2015 Poultry Science Association Inc.

Chemical and ruminal in vitro evaluation of Canadian canola meals produced over 4 years.

PubMed

Broderick, Glen A; Colombini, Stefania; Costa, Sara; Karsli, Mehmet A; Faciola, Antonio P

2016-10-01

To test the effects of year and processing plant on the nutritional value of canola meal (CM), 3 CM samples/yr were collected from each of 12 Canadian production plants over 4yr (total=144). Samples of CM were analyzed for differences in chemical composition and for in vitro ruminal protein degradability using the Michaelis-Menten inhibitor in vitro (MMIIV) method. In the MMIIV method, protein degradation rate (kd) was estimated by 2 methods: from net release (i.e., blank corrected) of (1) ammonia plus AA determined by o-phthaldialdehyde fluorescence (OPAF) assay or (2) ammonia, AA, plus oligopeptides determined by o-phthaldialdehyde absorbance (OPAA) assay; rumen-undegradable protein (RUP) was computed assuming passage rates of 0.16 and 0.06/h for, respectively, soluble and insoluble protein. Casein, solvent soybean meal (SSBM), and expeller soybean meal (ESBM) were included in all incubations as standard proteins. Differences among years and plants were assessed using the mixed procedures of SAS. Small but significant differences were found in CM among years for chemical composition, including N solubility; some of these differences may have been related to changes in our analytical methods over time. However, adjustment of degradation activity of individual in vitro incubations based on the mean degradation activity over all incubations yielded kd and RUP that did not differ by year using either assay. Simultaneously incubating CM samples from 2yr in the same in vitro runs confirmed that no year effects existed for kd or RUP. Differences existed in chemical composition of CM among the 12 processing plants over the 4yr of sample collection. Moreover, consistent differences in kd and RUP were observed among plants: kd ranged from 0.069 to 0.113/h (OPAA assay) and 0.075 to 0.120/h (OPAF assay), and RUP estimates ranged from 51 to 43% (OPAA assay) and 49 to 41% (OPAF assay). Regression of kd on insoluble N content of CM yielded correlation coefficients (R(2))=0.40 (OPAA assay) and 0.42 (OPAF assay), and regressions of kd on NDIN and N-fraction B3 yielded R(2)<0.02. Mean estimates from both OPAA and OPAF assays for casein, SSBM, ESBM, and CM were, respectively, kd=0.764, 0.161, 0.050, and 0.093/h and RUP=18, 33, 56, and 45%. A range of 8 percentage units from lowest to highest RUP suggests that substantial differences exist in metabolizable protein content of CM produced by different processing plants. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Quantitative evaluation of bone resorption activity of osteoclast-like cells by measuring calcium phosphate resorbing area using incubator-facilitated and video-enhanced microscopy.

PubMed

Morimoto, Yoshitaka; Hoshino, Hironobu; Sakurai, Takashi; Terakawa, Susumu; Nagano, Akira

2009-04-01

Quantitative evaluation of the ability of bone resorption activity in live osteoclast-like cells (OCLs) has not yet been reported on. In this study, we observed the sequential morphological change of OCLs and measured the resorbing calcium phosphate (CP) area made by OCLs alone and with the addition of elcatonin utilizing incubator facilitated video-enhanced microscopy. OCLs, which were obtained from a coculture of ddy-mouse osteoblastic cells and bone marrow cells, were cultured on CP-coated quartz cover slips. The CP-free area increased constantly in the OCLs alone, whereas it did not increase after the addition of elcatonin. This study showed that analysis of the resorbed areas under the OCL body using this method enables the sequential quantitative evaluation of the bone resorption activity and the effect of several therapeutic agents on bone resorption in vitro.

Influence of incubation temperature on biofilm formation and corrosion of carbon steel by Serratia marcescens

NASA Astrophysics Data System (ADS)

Harimawan, Ardiyan; Devianto, Hary; Kurniawan, Ignatius Chandra; Utomo, Josephine Christine

2017-01-01

Microbial induced corrosion (MIC) or biocorrosion is one type of corrosion, directly or indirectly influenced by microbial activities, by forming biofilm and adhering on the metal surface. When forming biofilm, the microorganisms can produce extracellular products which influence the cathodic and anodic reactions on metal surfaces. This will result in electrochemical changes in the interface between the biofilm and the metal surface, leading to corrosion and deterioration of the metal. MIC might be caused by various types of microorganism which leads to different corrosion mechanism and reaction kinetics. Furthermore, this process will also be influenced by various environmental conditions, such as pH and temperature. This research is aimed to determine the effect of incubation temperature on corrosion of carbon steel caused by Serratia marcescens in a mixture solution of synthetic seawater with Luria Bertani medium with a ratio of 4:1. The incubation was performed for 19 days with incubation temperature of 30, 37, and 50°C. The analyses of biofilm were conducted by total plate count (TPC), scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Biofilm was found to be evenly growth on the surface and increasing with increasing incubation temperature. It consists of functional group of alcohol, alkane, amine, nitro, sulfate, carboxylic acid, and polysulfide. The analyses of the corrosion were conducted by gravimetric and X-ray diffraction (XRD). Higher incubation temperature was found to increase the corrosion rate. However, the corrosion products were not detected by XRD analysis.

Testing of mechanical ventilators and infant incubators in healthcare institutions.

PubMed

Badnjevic, Almir; Gurbeta, Lejla; Jimenez, Elvira Ruiz; Iadanza, Ernesto

2017-01-01

The medical device industry has grown rapidly and incessantly over the past century. The sophistication and complexity of the designed instrumentation is nowadays rising and, with it, has also increased the need to develop some better, more effective and efficient maintenance processes, as part of the safety and performance requirements. This paper presents the results of performance tests conducted on 50 mechanical ventilators and 50 infant incubators used in various public healthcare institutions. Testing was conducted in accordance to safety and performance requirements stated in relevant international standards, directives and legal metrology policies. Testing of output parameters for mechanical ventilators was performed in 4 measuring points while testing of output parameters for infant incubators was performed in 7 measuring points for each infant incubator. As performance criteria, relative error of output parameters for mechanical ventilators and absolute error of output parameters for infant incubators was calculated. The ranges of permissible error, for both groups of devices, are regulated by the Rules on Metrological and Technical Requirements published in the Official Gazette of Bosnia and Herzegovina No. 75/14, which are defined based on international recommendations, standards and guidelines. All ventilators and incubators were tested by etalons calibrated in an ISO 17025 accredited laboratory, which provides compliance to international standards for all measured parameters.The results show that 30% of the tested medical devices are not operating properly and should be serviced, recalibrated and/or removed from daily application.

Physical and chemical properties of biochars co-composted with biowastes and incubated with a chicken litter compost.

PubMed

Khan, Naser; Clark, Ian; Sánchez-Monedero, Miguel A; Shea, Syd; Meier, Sebastian; Qi, Fangjie; Kookana, Rai S; Bolan, Nanthi

2016-01-01

Two experiments were conducted where three biochars, made from macadamia nutshell (MS), hardwood shaving (WS) and chicken litter (CL), were co-composted with chicken manure and sawdust, and also incubated with a chicken litter based commercial compost. Biochars were added at the rates of 5% and 10% in the co-composting and 10% and 20% in the incubation experiment. The rates of biochar had no consistent effect on the change in element contents of composted- or incubated-biochars. The biochar C demonstrated recalcitrance in both composting and incubation systems. Composting increased the CEC of biochars probably due to thermophilic oxidation. The increases in CEC of WS and CL were 6.5 and 2.2 times, respectively, for composting. Translocation of elements, between biochar and compost medium, occurred in both directions. In most cases, biochars gained elements under the influence of positive difference of concentrations (i.e., when compost medium had higher concentration of elements than biochar), while in some cases they lost elements despite a positive difference. Biochar lost some elements (WS: B; CL: B, Mg and S) under the influence of negative difference of concentrations. Some biochars showed strong affinity for B, C, N and S: the concentration of these elements gained by biochars surpassed the concentration in the respective composting medium. The material difference in the biochars did not have influence on N retention: all three netbag-biochars increased their N content. The cost of production of biochar-compost will be lower in co-composting than incubation, which involves two separate processes, i.e., composting and subsequent incubation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Drug release from porous silicon for stable neural interface

NASA Astrophysics Data System (ADS)

Sun, Tao; Tsang, Wei Mong; Park, Woo-Tae

2014-02-01

70 μm-thick porous Si (PSi) layer with the pore size of 11.1 ± 7.6 nm was formed on an 8-in. Si wafer via an anodization process for the microfabrication of a microelectrode to record neural signals. To reduce host tissue responses to the microelectrode and achieve a stable neural interface, water-soluble dexamethesone (Dex) was loaded into the PSi via incubation with the drug solution overnight. After the drug loading process, the pore size of PSi reduced to 4.7 ± 2.6 nm on the basis of scanning electron microscopic (SEM) images, while its wettability was remarkably enhanced. Fluorescence images demonstrated that Dex was loaded into the porous structure of the PSi. Degradation rate of the PSi was investigated by incubation in distilled water for 21 days. Moreover, the drug release profile of the Dex-loaded PSi was a combination of an initial burst release and subsequent sustained release. To evaluate cellular responses to the drug release from the PSi, primary astrocytes were seeded on the surface of samples. After 2 days of culture, the Dex-loaded PSi could not only moderately prevent astrocyte adhesion in comparison with Si, but also more effectively suppress the activation of primary astrocytes than unloaded PSi due to the drug release. Therefore, it might be an effective method to reduce host tissue responses and stabilize the quality of the recorded neural signal by means of loading drugs into the PSi component of the microelectrode.

Bioavailability of particle-associated Se to the bivalve Potamocorbula amurensis

USGS Publications Warehouse

Schlekat, C.E.; Dowdle, P.R.; Lee, B.-G.; Luoma, S.N.; Oremland, R.S.

2000-01-01

Elemental selenium, Se(0), is a prevalent chemical form in sediments, but little is known about its bioavailability. We evaluated the bioavailability of two forms of Se(0) by generating radioisotopic 75Se(0) through bacterial dissimilatory reduction of 75SeO32- by pure bacterial cultures (SES) and by an anaerobic sediment microbial consortium (SED). A third form was generated by reducing 75SeO32- with ascorbic acid (AA). Speciation determinations showed that AA and SES were >90% Se(0), but SED showed a mixture of Se(0), selenoanions, and a residual fraction. Pulse-chase techniques were used to measure assimilation efficiencies (AE) of these particulate Se forms by the bivalve Potamocorbula amurensis. Mean AE values were 3 ?? 2% for AA, 7 ?? 1% for SES, and 28 ?? 15% for SED, showing that the bioavailability of reduced, particle-associated Se is dependent upon its origin. To determine if oxidative microbial processes increased Se transfer, SES 75Se(0) was incubated with an aerobic sediment microbial consortium. After 113 d of incubation, 36% of SES Se(0) was oxidized to SeO32-. Assimilation of total particulate Se was unaffected however (mean AE = 5.5%). The mean AE from the diatom Phaeodactylum tricornutum was 58 ?? 8%, verifying the importance of Se associated with biogenic particles. Speciation and AE results from SED suggest that selenoanion reduction in wetlands and estuaries produces biologically available reduced selenium.Elemental selenium, Se(0), is a prevalent chemical form in sediments, but little is known about its bioavailability. We evaluated the bioavailability of two forms of Se(0) by generating radioisotopic 75Se(0) through bacterial dissimilatory reduction of 75SeO32- by pure bacterial cultures (SES) and by an anaerobic sediment microbial consortium (SED). A third form was generated by reducing 75SeO32 with ascorbic acid (AA). Speciation determinations showed that AA and SES were > 90% Se(0), but SED showed a mixture of Se(0), selenoanions, and a residual fraction. Pulse-chase techniques were used to measure assimilation efficiencies (AE) of these particulate Se forms by the bivalve Potamocorbula amurensis. Mean AE values were 3 ?? 2% for AA, 7 ?? 1% for SES, and 28 ?? 15% for SED, showing that the bioavailability of reduced, particle-associated Se is dependent upon its origin. To determine if oxidative microbial processes increased Se transfer, SES 75Se(0) was incubated with an aerobic sediment microbial consortium. After 113 d of incubation, 36% of SES Se(0) was oxidized to SeO32-. Assimilation of total particulate Se was unaffected however (mean AE = 5.5%). The mean AE from the diatom Phaeodactylum tricornutum was 58 ?? 8%, verifying the importance of Se associated with biogenic particles. Speciation and AE results from SED suggest that selenoanion reduction in wetlands and estuaries produces biologically available reduced selenium.

Metabolic Characterization of Acutely Isolated Hippocampal and Cerebral Cortical Slices Using [U-13C]Glucose and [1,2-13C]Acetate as Substrates.

PubMed

McNair, Laura F; Kornfelt, Rasmus; Walls, Anne B; Andersen, Jens V; Aldana, Blanca I; Nissen, Jakob D; Schousboe, Arne; Waagepetersen, Helle S

2017-03-01

Brain slice preparations from rats, mice and guinea pigs have served as important tools for studies of neurotransmission and metabolism. While hippocampal slices routinely have been used for electrophysiology studies, metabolic processes have mostly been studied in cerebral cortical slices. Few comparative characterization studies exist for acute hippocampal and cerebral cortical slices, hence, the aim of the current study was to characterize and compare glucose and acetate metabolism in these slice preparations in a newly established incubation design. Cerebral cortical and hippocampal slices prepared from 16 to 18-week-old mice were incubated for 15-90 min with unlabeled glucose in combination with [U- 13 C]glucose or [1,2- 13 C]acetate. Our newly developed incubation apparatus allows accurate control of temperature and is designed to avoid evaporation of the incubation medium. Subsequent to incubation, slices were extracted and extracts analyzed for 13 C-labeling (%) and total amino acid contents (µmol/mg protein) using gas chromatography-mass spectrometry and high performance liquid chromatography, respectively. Release of lactate from the slices was quantified by analysis of the incubation media. Based on the measured 13 C-labeling (%), total amino acid contents and relative activity of metabolic enzymes/pathways, we conclude that the slice preparations in the current incubation apparatus exhibited a high degree of metabolic integrity. Comparison of 13 C-labeling observed with [U- 13 C]glucose in slices from cerebral cortex and hippocampus revealed no significant regional differences regarding glycolytic or total TCA cycle activities. On the contrary, results from the incubations with [1,2- 13 C]acetate suggest a higher capacity of the astrocytic TCA cycle in hippocampus compared to cerebral cortex. Finally, we propose a new approach for assessing compartmentation of metabolite pools between astrocytes and neurons using 13 C-labeling (%) data obtained from mass spectrometry. Based on this approach we suggest that cellular metabolic compartmentation in hippocampus and cerebral cortex is very similar.

Fundamental Space Biology-1: HHR and Incubator for ISS Space Life Sciences

NASA Astrophysics Data System (ADS)

Kirven-Brooks, M.; Fahlen, T.; Sato, K.; Reiss-Bubenheim, D.

The Space Station Biological Research Project (SSBRP) is developing an Incubator and a Habitat Holding Rack (HHR) to support life science experiments aboard the International Space Station (ISS). The HHR provides for cooling and power needs, and supports data transfer (including telemetry, commanding, video processing, Ethernet), video compression, and data and command storage). The Incubator is a habitat that provides for controlled temperature between +4 C and +45 C and air circulation. It has a set of connector ports for power, analog and digital sensors, and video pass-through to support experiment-unique hardware within the Incubator specimen chamber. The Incubator exchanges air with the ISS cabin. The Fundamental Space Biology-1 (FSB-1) Project will be delivering, the HHR and two Incubators to ISS. The two inaugural experiments to be conducted on ISS using this hardware will investigate the biological effects of the space environment on two model organisms, Saccharomyces cerevisiae (S. cerevisiae; yeast) and Caenorhabditis elegans (C. elegans; nematode). The {M}odel {Y}east {C}ultures {o}n {S}tation (MYCOS) experiment will support examination of the effect of microgravity and cosmic radiation on yeast biology. In the second series of experiments during the same increment, the effects of microgravity and space environment radiation on C. elegans will be examined. The {F}undamental Space Biology {I}ncubator {E}xperiment {R}esearch using {C}. {e}legans (FIERCE) study is designed to support a long duration, multi-generational study of nematodes. FIERCE on-orbit science operations will include video monitoring, sub-culturing and periodic fixation and freezing of samples. For both experiments, investigators will be solicited via an International Space Life Sciences Research Announcement. In the near future, the Centrifuge Accommodation Module will be delivered to ISS, which will house the SSBRP 2.5 m Centrifuge Rotor. The Incubator can be placed onto the Centrifuge Rotor, which is capable of supporting variable gravity experiments from microgravity through 2g.

A comparison of the ability of Bellucia dichotoma Cogn. (Melastomataceae) extract to inhibit the local effects of Bothrops atrox venom when pre-incubated and when used according to traditional methods.

PubMed

Mourão de Moura, Valéria; Serra Bezerra, Adrielle N; Veras Mourão, Rosa Helena; Varjão Lameiras, Juliana L; Almeida Raposo, Juliana D; Luckwu de Sousa, Rafael; Boechat, Antônio Luiz; Bezerra de Oliveira, Ricardo; de Menezes Chalkidis, Hipocrátes; Dos-Santos, Maria Cristina

2014-07-01

Bellucia dichotoma Cogn. (Melastomataceae) is one of various plant species used in folk medicine in the west of the state of Pará, Brazil, to treat snake bites. Many studies have been carried out to evaluate the effectiveness of anti-snake bite plants, but few of these use the same preparation methods and doses as those traditionally used by the local populations. This study therefore compared inhibition of the main local effects of B. atrox venom (BaV) by aqueous extract of B. dichotoma (AEBd) administered according to traditional methods and pre-incubated with BaV). The concentrations of phenolic compounds (tannins and flavonoids) in AEBd were determined by colorimetric assays. The effectiveness of AEBd in inhibiting the hemorrhagic and edematogenic activities of BaV was evaluated in mice in four different experimental in vivo protocols: (1) pre-incubation (venom:extract, w/w); (2) pre-treatment (p.o.); (3) post-treatment (p.o.); and (4) AEBd (p.o.) in combination with Bothrops antivenom (BA) (i.v.). To assess in vitro inhibition of BaV phospholipase A₂ activity, the pre-incubation method or incorporation of AEBd or BA in agarose gels were used. The effect of AEBd on BaV was determined by SDS-PAGE, zymography and Western blot. Colorimetric assays revealed higher concentrations of (condensed and hydrolyzable) tannins than flavonoids in AEBd. Hemorrhagic activity was completely inhibited using the pre-incubation protocol. However, with pre-treatment there was no significant inhibition for the concentrations tested, and with the post-treatment only the 725 mg/kg dose of AEBd was able to inhibit 40.5% (p = 0.001) of the hemorrhagic activity of BaV. Phospholipase A₂ activity was only inhibited when AEBd was pre-incubated with BaV. BaV-induced edema was completely inhibited with pre-incubation (p < 0.05) and significantly reduced (p < 0.05) with pre- and post-treatment (p.o.) for the concentrations tested. The reduction in local edema was even greater when AEBd was administered in combination with BA. The SDS-PAGE profiles showed that several of the BaV protein (SDS-PAGE) and enzyme (zymography) bands were not detected when the venom was pre-incubated, and Western blot revealed that this was not caused by the AEBd enzymes observed in the zymogram. The "pseudo inhibition" observed after pre-incubation in this study may be due to the presence of tannins in the extract, which could act as chelating agents, removing metalloproteins and Ca²⁺ ions and thus inhibiting hemorrhagin and PLA₂ activity. However, when administered according to traditional methods, B. dichotoma extract was effective in blocking BaV-induced edematogenic activity and had an additional effect on inhibition of this activity by BA. Copyright © 2014 Elsevier Ltd. All rights reserved.

Cu isotope fractionation response to oxidative stress in a hepatic cell line studied using multi-collector ICP-mass spectrometry.

PubMed

Flórez, María R; Costas-Rodríguez, Marta; Grootaert, Charlotte; Van Camp, John; Vanhaecke, Frank

2018-03-01

Reactive oxygen species (ROS) are generated in biological processes involving electron transfer reactions and can act in a beneficial or deleterious way. When intracellular ROS levels exceed the cell's anti-oxidant capacity, oxidative stress occurs. In this work, Cu isotope fractionation was evaluated in HepG2 cells under oxidative stress conditions attained in various ways. HepG2 is a well-characterised human hepatoblastoma cell line adapted to grow under high oxidative stress conditions. During a pre-incubation stage, cells were exposed to a non-toxic concentration of Cu for 24 h. Subsequently, the medium was replaced and cells were exposed to one of three different external stressors: H 2 O 2 , tumour necrosis factor α (TNFα) or UV radiation. The isotopic composition of the intracellular Cu was determined by multi-collector ICP-mass spectrometry to evaluate the isotope fractionation accompanying Cu fluxes between cells and culture medium. For half of these setups, the pre-incubation solution also contained N-acetyl-cysteine (NAC) as an anti-oxidant to evaluate its protective effect against oxidative stress via its influence on the extent of Cu isotope fractionation. Oxidative stress caused the intracellular Cu isotopic composition to be heavier compared to that in untreated control cells. The H 2 O 2 and TNFα exposures rendered similar results, comparable to those obtained after mild UV exposure. The heaviest Cu isotopic composition was observed under the strongest oxidative conditions tested, i.e., when the cell surfaces were directly exposed to UV radiation without apical medium and in absence of NAC. NAC mitigated the extent of isotope fractionation in all cases.

Short-term effects of tidal flooding on soil nitrogen mineralization in a Chinese tidal salt marsh

NASA Astrophysics Data System (ADS)

Gao, Haifeng; Bai, Junhong; Deng, Xiaoya; Lu, Qiongqiong; Ye, Xiaofei

2018-02-01

Tidal flooding is an important control of nitrogen biogeochemistry in wetland ecosystems of Yellow River Delta, China. Variations in hydrology could change soil redox dynamics and conditions for microorganisms living. A tidal simulation experiment was designed to extract tidal flooding effect on nitrogen mineralization of salt marsh soil. Inorganic nitrogen and relevant enzyme were measured during the 20-day incubation period. Considering the variation of both inorganic N and enzymes, nitrogen mineralization process in tidal salt marsh could be divided into 2 phases of short term response and longtime adaption by around 12th incubation day as the inflection point. Soil ammonium nitrogen (NH4+-N) and volatilized ammonia (NH3) occupied the mineralization process since nitrate nitrogen (NO3--N) was not detected over whole incubation period. NH4+-N varied fluctuant and increased significantly after 12 day's incubation. Released NH3 reached to peak value of 14.24 mg m-2 d-1 at the inflection point and declined thereafter. Inorganic nitrogen released according to net nitrogen mineralization rate (RM) under the tidal flooding condition without plant uptake except first 2 days. However, during the transitional period of 6-12 days, RM decreased notably to almost 0 and increased again after inflection point with the value of 0.182 mg kg-1 d-1. It might be due to the change of microbial composition and function when soil shifted from oxic to anoxic, which were reflected by arylamidase, urease and fluorescein diacetate. Fluorescein diacetate hydrolysis and arylamidase had the similar variation of U style with decreasing activities before 12 days' incubation. All the enzymes measured in this experiment increased after inflection point. Whereas, urease activity kept constant from 2 to 12 days. Alternant oxidation reduction condition would increase N loss through denitrification and ammonia volatilization during the transitional period, while more inorganic nitrogen would be available in reductive environment of long-term tidal flooding. Therefore, hydrological process regulation has great influence on nitrogen cycling and further influence on wetland productivity.

Neurotoxicological effects of nicotine on the embryonic development of cerebellar cortex of chick embryo during various stages of incubation.

PubMed

El-Beltagy, Abd El-Fattah B M; Abou-El-Naga, Amoura M; Sabry, Dalia M

2015-10-01

Long-acting nicotine is known to exert pathological effects on almost all tissues including the cerebellar cortex. The present work was designed to elucidate the effect of nicotine on the development of cerebellar cortex of chick embryo during incubation period. The fertilized eggs of hen (Gallus gallus domesticus) were injected into the air space by a single dose of long acting nicotine (1.6 mg/kg/egg) at the 4th day of incubation. The embryos were taken out of the eggs on days 8, 12 and 16 of incubation. The cerebellum of the control and treated embryos at above ages were processed for histopathological examination. The TEM were examined at 16th day of incubation. The results of the present study revealed that, exposure to long-acting nicotine markedly influence the histogenesis of cerebellar cortex of chick embryo during the incubation period. At 8th day of incubation, nicotine delayed the differentiation of the cerebellar analge; especially the external granular layer (EGL) and inner cortical layer (ICL). Furthermore, at 12th day of incubation, the cerebellar foliation was irregular and the Purkinje cells not recognized. By 16th day of incubation, the cerebellar foliations were irregular with interrupted cerebellar cortex and irregular arrangement of Purkinje cells. Immunohistochemical analysis for antibody P53 protein revealed that the cerebellar cortex in all stages of nicotine treated groups possessed a moderate to weak reaction for P53 protein however; this reaction was markedly stronger in the cerebellar cortex of control groups. Moreover, the flow cytometric analysis confirmed that the percentage of apoptosis in control group was significantly higher compared with that of nicotine treated group. At the TEM level, the cerebellar Purkinje cells of 16th day of treated groups showed multiple subcellular alterations in compared with those of the corresponding control group. Such changes represented by appearing of vacuolated mitochondria, cisternal fragmentation of RER, irregular grooves of Golgi tubules. Also, multiple cytoplasmic vacuoles and aggregation of Nissl granules were recorded around pyknotic nucleus. Copyright © 2015 Elsevier Ltd. All rights reserved.

An improved glucose transport assay system for isolated mouse skeletal muscle tissues.

PubMed

Inagaki, Akiko; Maruo, Kanoko; Furuichi, Yasuro; Miyatake, Shouta; Tamura, Kotaro; Fujii, Nobuharu L; Manabe, Yasuko

2016-07-18

There is a growing demand for a system in the field of sarcopenia and diabetes research that could be used to evaluate the effects of functional food ingredients that enhance muscle mass/contractile force or muscle glucose uptake. In this study, we developed a new type of in vitro muscle incubation system that systemizes an apparatus for muscle incubation, using an electrode, a transducer, an incubator, and a pulse generator in a compact design. The new system enables us to analyze the muscle force stimulated by the electric pulses and glucose uptake during contraction and it may thus be a useful tool for analyzing the metabolic changes that occur during muscle contraction. The system may also contribute to the assessments of new food ingredients that act directly on skeletal muscle in the treatment of sarcopenia and diabetes.

[Effects of soil properties on the stabilization process of cadmium in Cd alone and Cd-Pb contaminated soils].

PubMed

Wu, Man; Xu, Ming-Gang; Zhang, Wen-Ju; Wu, Hai-Wen

2012-07-01

In order to clarify the effects of soil properties on the stabilization process of the cadmium (Cd) added, 11 different soils were collected and incubated under a moisture content of 65%-70% at 25 degrees C. The changes of available Cd contents with incubation time (in 360 days) in Cd and Cd-Pb contaminated treatments were determined. The stabilization process was simulated using dynamic equations. The results showed that after 1.0 mg x kg(-1) Cd or 500 mg x kg(-1) Pb + 1.0 mg x kg(-1) Cd were added into the soil, the available Cd content decreased rapidly during the first 15 days, and then the decreasing rate slowed down, with an equilibrium content reached after 60 days' incubation. In Cd-Pb contaminated soils, the presence of Pb increased the content of available Cd. The stabilization process of Cd could be well described by the second-order equation and the first order exponential decay; meanwhile, dynamic parameters including equilibrium content and stabilization velocity were used to characterize the stabilization process of Cd. These two key dynamic parameters were significantly affected by soil properties. Correlation analysis and stepwise regression suggested that high pH and high cation exchange capacity (CEC) significantly retarded the availability of Cd. High pH had the paramount effect on the equilibrium content. The stabilization velocity of Cd was influenced by the soil texture. It took shorter time for Cd to get stabilized in sandy soil than in the clay.

The effect of isolation and culture methods on epithelial stem cell populations and their progeny-toward an improved cell expansion protocol for clinical application.

PubMed

Lenihan, Catherine; Rogers, Caroline; Metcalfe, Anthony D; Martin, Yella H

2014-12-01

The use of cultured epithelial keratinocytes in the treatment of burns and skin graft donor sites is well established in clinical practice. The most widely used culture method for clinical use was originally developed by Rheinwald and Green 40 years ago. This system uses irradiated mouse dermal fibroblasts as a feeder cell layer to promote keratinocyte growth, a process that is costly and labor-intensive for health care providers. The medium formulation contains several components of animal origin, which pose further safety risks for patients. Improvements and simplification in the culturing process would lead to clear advantages: improved safety through reduction of xenobiotic components and reduction in cost for health care providers by dispensing with feeder cells. We compared the Rheinwald and Green method to culture in three commercially available, feeder-free media systems with defined/absent components of animal origin. During the isolation process, short incubation times in high-strength trypsin resulted in increased numbers of liberated keratinocyte stem cells compared with longer incubation times. All three commercially available media tested in this study could support the expansion of keratinocytes, with phenotypes comparable to cells expanded using the established Rheinwald and Green method. Growth rates varied, with two of the media displaying comparable growth rates, whereas the third was significantly slower. Our study demonstrates the suitability of such feeder-free media systems in clinical use. It further outlines a range of techniques to evaluate keratinocyte phenotype when assessing the suitability of cells for clinical application. Copyright © 2014 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

High Temperature and Salinity Enhance Soil Nitrogen Mineralization in a Tidal Freshwater Marsh

PubMed Central

Gao, Haifeng; Bai, Junhong; He, Xinhua; Zhao, Qingqing; Lu, Qiongqiong; Wang, Junjing

2014-01-01

Soil nitrogen (N) mineralization in wetlands is sensitive to various environmental factors. To compare the effects of salinity and temperature on N mineralization, wetland soils from a tidal freshwater marsh locating in the Yellow River Delta was incubated over a 48-d anaerobic incubation period under four salinity concentrations (0, 10, 20 and 35‰) and four temperature levels (10, 20, 30 and 40°C). The results suggested that accumulated ammonium nitrogen (NH4 +-N) increased with increasing incubation time under all salinity concentrations. Higher temperatures and salinities significantly enhanced soil N mineralization except for a short-term (≈10 days) inhibiting effect found under 35‰ salinity. The incubation time, temperature, salinity and their interactions exhibited significant effects on N mineralization (P<0.001) except the interactive effect of salinity and temperature (P>0.05), while temperature exhibited the greatest effect (P<0.001). Meanwhile, N mineralization processes were simulated using both an effective accumulated temperature model and a one-pool model. Both models fit well with the simulation of soil N mineralization process in the coastal freshwater wetlands under a range of 30 to 40°C (R2 = 0.88–0.99, P<0.01). Our results indicated that an enhanced NH4 +-N release with increasing temperature and salinity deriving from the projected global warming could have profound effects on nutrient cycling in coastal wetland ecosystems. PMID:24733366

Rapid Identification of Microorganisms from Positive Blood Culture by MALDI-TOF MS After Short-Term Incubation on Solid Medium.

PubMed

Curtoni, Antonio; Cipriani, Raffaella; Marra, Elisa Simona; Barbui, Anna Maria; Cavallo, Rossana; Costa, Cristina

2017-01-01

Matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is a useful tool for rapid identification of microorganisms. Unfortunately, its direct application to positive blood culture is still lacking standardized procedures. In this study, we evaluated an easy- and rapid-to-perform protocol for MALDI-TOF MS direct identification of microorganisms from positive blood culture after a short-term incubation on solid medium. This protocol was used to evaluate direct identification of microorganisms from 162 positive monomicrobial blood cultures; at different incubation times (3, 5, 24 h), MALDI-TOF MS assay was performed from the growing microorganism patina. Overall, MALDI-TOF MS concordance with conventional methods at species level was 60.5, 80.2, and 93.8% at 3, 5, and 24 h, respectively. Considering only bacteria, the identification performances at species level were 64.1, 85.0, and 94.1% at 3, 5, and 24 h, respectively. This protocol applied to a commercially available MS typing system may represent, a fast and powerful diagnostic tool for pathogen direct identification and for a promptly and pathogen-driven antimicrobial therapy in selected cases.

Evaluation of incubation time for Microsporum canis dermatophyte cultures.

PubMed

Stuntebeck, Rebecca; Moriello, Karen A; Verbrugge, Maria

2017-09-01

Objectives The goal of this study was to determine how frequently Microsporum canis was isolated after 1, 2 and 3 weeks of incubation on dermatophyte culture medium either from untreated cats or cats during treatment. Methods This was an observational retrospective study. Toothbrush fungal culture results were examined from two data pools: untreated cats with suspect skin lesions and weekly fungal cultures from cats being treated for dermatophytosis. Results Results from 13,772 fungal cultures were reviewed and 2876 (20.9%) were positive for M canis. Of these, 2800 were confirmed as positive within 14 days of incubation and only 76 (2.6%) required >14 days for confirmation of M canis. In pretreatment specimens, 98.2% (1057/1076) of M canis isolates were recovered within 14 days of incubation in specimens from cats not known to have received prior antifungal treatment. For cats receiving treatment, 96.8% (1743/1800) of M canis isolates were recovered within 14 days of incubation. Of the 57 cultures that required >14 days for finalization, 21 required extra incubation time because cultures were grossly abnormal, 12 had concurrent contaminant growth delaying microscopic confirmation and 24 had no growth in the first 14 days. Of these 24, 19 had 1-2 colony-forming units (cfu)/plate and the remaining five plates had 5 to >10 cfu/plate, all with abnormal morphology. Conclusions and relevance The findings of this study show that it is not necessary to hold pretreatment or post-treatment fungal cultures for 21 days before finalizing cultures for no growth. Growth requiring >14 days had grossly abnormal morphology.

Nest survival is influenced by parental behaviour and heterospecifics in a mixed-species colony

USGS Publications Warehouse

Brussee, Brianne E.; Coates, Peter S.; Hothem, Roger L.; Howe, Kristy; Casazza, Michael L.; Eadie, John M.

2016-01-01

Studies of avian nest success often focus on examining influences of variation in environmental and seasonal factors. However, in-depth evaluations can also incorporate variation in individual incubation behaviour to further advance our understanding of avian reproductive ecology. We examined these relationships in colonially nesting Black-crowned Night-Herons Nycticorax nycticorax using intensive video-monitoring methods to quantify incubation behaviours. We modelled nest survival as a function of both extrinsic factors and incubation behaviours over a 3-year period (2010–12) on Alcatraz Island, USA. Model-averaged parameter estimates indicated that nest survival increased as a function of greater incubation constancy (% of time spent incubating eggs within a 24-h period), and average daily precipitation throughout the nesting stage. Common Ravens Corvus corax are the only known nest predator of Night-Herons on Alcatraz Island, as on many other coastal Pacific islands. We also investigated the effects of heterospecific nesting of California Gulls Larus californicus and Western Gulls Larus occidentalis in a mixed-species colony with Night-Herons, based on nesting proximity data collected over a 2-year period (2011–12). This second analysis indicated that, in addition to incubation behaviours, nesting heterospecifics are an important factor for explaining variation in Night-Heron nest survival. However, contrary to our original expectation, we found that Night-Herons experienced increased nest survival with increasing distance from gull colony boundaries. These results may apply to other areas with multiple colonial nesting species and similar predator communities and climatic patterns.

Characterization of incubation experiments and development of an enrichment culture capable of ammonium oxidation under iron reducing conditions

NASA Astrophysics Data System (ADS)

Huang, S.; Jaffé, P. R.

2014-08-01

Incubation experiments were conducted using soil samples from a forested riparian wetland where we have previously observed anaerobic ammonium oxidation coupled to iron reduction. Production of both nitrite and ferrous iron were measured repeatedly during incubations when the soil slurry was supplied with either ferrihydrite or goethite and ammonium chloride. Significant changes in the microbial community were observed after 180 days of incubation as well as in a continuous flow membrane reactor, using 16S rRNA gene PCR-denaturing gradient gel electrophoresis, 454-pyrosequencing, and real-time quantitative PCR analysis. We believe that one of the dominant microbial species in our system (an uncultured Acidimicrobiaceae bacterium A6), belonging to the Acidimicrobiaceae family, whose closest cultivated relative is Ferrimicrobium acidiphilum (with 92% identity) and Acidimicrobium ferrooxidans (with 90% identity), might play a key role in this anaerobic biological process that uses ferric iron as an electron acceptor while oxidizing ammonium to nitrite. After ammonium was oxidized to nitrite, nitrogen loss proceeded via denitrification and/or anammox.

Significant increase in cultivation of Gardnerella vaginalis, Alloscardovia omnicolens, Actinotignum schaalii, and Actinomyces spp. in urine samples with total laboratory automation.

PubMed

Klein, Sabrina; Nurjadi, Dennis; Horner, Susanne; Heeg, Klaus; Zimmermann, Stefan; Burckhardt, Irene

2018-04-13

While total laboratory automation (TLA) is well established in laboratory medicine, only a few microbiological laboratories are using TLA systems. Especially in terms of speed and accuracy, working with TLA is expected to be superior to conventional microbiology. We compared in total 35,564 microbiological urine cultures with and without incubation and processing with BD Kiestra TLA for a 6-month period each retrospectively. Sixteen thousand three hundred thirty-eight urine samples were analyzed in the pre-TLA period and 19,226 with TLA. Sixty-two percent (n = 10,101/16338) of the cultures processed without TLA and 68% (n = 13,102/19226) of the cultures processed with TLA showed growth. There were significantly more samples with two or more species per sample and with low numbers of colony forming units (CFU) after incubation with TLA. Regarding the type of bacteria, there were comparable amounts of Enterobacteriaceae in the samples, slightly less non-fermenting Gram-negative bacteria, but significantly more Gram-positive cocci, and Gram-positive rods. Especially Alloscardivia omnicolens, Gardnerella vaginalis, Actinomyces spp., and Actinotignum schaalii were significantly more abundant in the samples incubated and processed with TLA. The time to report was significantly lower in the TLA processed samples by 1.5 h. We provide the first report in Europe of a large number of urine samples processed with TLA. TLA showed enhanced growth of non-classical and rarely cultured bacteria from urine samples. Our findings suggest that previously underestimated bacteria may be relevant pathogens for urinary tract infections. Further studies are needed to confirm our findings.

Curated Collections for Educators: Five Key Papers about Program Evaluation.

PubMed

Thoma, Brent; Gottlieb, Michael; Boysen-Osborn, Megan; King, Andrew; Quinn, Antonia; Krzyzaniak, Sara; Pineda, Nicolas; Yarris, Lalena M; Chan, Teresa

2017-05-04

The evaluation of educational programs has become an expected part of medical education. At some point, all medical educators will need to critically evaluate the programs that they deliver. However, the evaluation of educational programs requires a very different skillset than teaching. In this article, we aim to identify and summarize key papers that would be helpful for faculty members interested in exploring program evaluation. In November of 2016, the 2015-2016 Academic life in emergency medicine (ALiEM) Faculty Incubator program highlighted key papers in a discussion of program evaluation. This list of papers was augmented with suggestions by guest experts and by an open call on Twitter. This resulted in a list of 30 papers on program evaluation. Our authorship group then engaged in a process akin to a Delphi study to build consensus on the most important papers about program evaluation for medical education faculty. We present our group's top five most highly rated papers on program evaluation. We also summarize these papers with respect to their relevance to junior medical education faculty members and faculty developers. Program evaluation is challenging. The described papers will be informative for junior faculty members as they aim to design literature-informed evaluations for their educational programs.

Curated Collections for Educators: Five Key Papers about Program Evaluation

PubMed Central

Gottlieb, Michael; Boysen-Osborn, Megan; King, Andrew; Quinn, Antonia; Krzyzaniak, Sara; Pineda, Nicolas; Yarris, Lalena M; Chan, Teresa

2017-01-01

The evaluation of educational programs has become an expected part of medical education. At some point, all medical educators will need to critically evaluate the programs that they deliver. However, the evaluation of educational programs requires a very different skillset than teaching. In this article, we aim to identify and summarize key papers that would be helpful for faculty members interested in exploring program evaluation. In November of 2016, the 2015-2016 Academic life in emergency medicine (ALiEM) Faculty Incubator program highlighted key papers in a discussion of program evaluation. This list of papers was augmented with suggestions by guest experts and by an open call on Twitter. This resulted in a list of 30 papers on program evaluation. Our authorship group then engaged in a process akin to a Delphi study to build consensus on the most important papers about program evaluation for medical education faculty. We present our group’s top five most highly rated papers on program evaluation. We also summarize these papers with respect to their relevance to junior medical education faculty members and faculty developers. Program evaluation is challenging. The described papers will be informative for junior faculty members as they aim to design literature-informed evaluations for their educational programs. PMID:28589073

In vitro effects of zinc, D-aspartic acid, and coenzyme-Q10 on sperm function.

PubMed

Giacone, Filippo; Condorelli, Rosita A; Mongioì, Laura M; Bullara, Valentina; La Vignera, Sandro; Calogero, Aldo E

2017-05-01

Reactive oxygen species favor reproductive processes at low concentrations, but damage spermatozoa and decrease their fertilizing capacity at high concentrations. During infection and/or inflammation of the accessory sex glands reactive oxygen species overproduction may occur which, in turn, may negatively impact on sperm motility, sperm DNA fragmentation, and lipid peroxidation. A number of nutraceutical formulations containing antioxidant molecules have been developed to counteract the deleterious effects of the oxidative stress. A recent formulation containing zinc, D-aspartic acid, and coenzyme-Q10 is present in the pharmaceutical market. Based on these premises, the aim of the present study was to evaluate the effects of this combination on spermatozoa in vitro. The study was conducted on 24 men (32.2 ± 5.5 years): 12 normozoospermic men and 12 asthenozoospermic patients. Spermatozoa from each sample were divided into two control aliquots (aliquot A and B) and an aliquot incubated with zinc, D-aspartic acid, and coenzyme-Q10 (aliquot C). After 3 h of incubation, the following parameters were evaluated: progressive motility, number of spermatozoa with progressive motility recovered after swim-up, lipid peroxidation, and DNA fragmentation. Incubation with zinc, D-aspartic acid, and coenzyme-Q10 maintained sperm motility in normozoospermic men (37.7 ± 1.2 % vs. 35.8 ± 2.3 % at time zero) and improved it significantly in asthenozoospermic patients (26.5 ± 1.9 % vs. 18.8 ± 2.0 % at time zero) (p < 0.01). This resulted in a significantly higher (p < 0.01) number of spermatozoa with progressive motility recovered after swim-up in both normozospermic men (4.1 ± 0.9 vs. 3.3 ± 1.0 millions) and asthenozooseprmic patients (3.2 ± 0.8 vs. 1.6 ± 0.5 millions). Finally, a statistically significant lower sperm lipid peroxidation was found after incubation with zinc, D-aspartic acid, and coenzyme-Q10 (p < 0.05) in both normozospermic men (1.0 ± 0.4 % vs. 2.4 ± 0.9 %) and asthenozooseprmic patients (0.2 ± 0.1 % vs. 0.6 ± 0.2 %). No statistically significant effect was observed on sperm DNA fragmentation. This nutraceutical formulation may be indicated in vitro during the separation of the spermatozoa in the assisted reproduction techniques, during which the spermatozoa undergo an increased oxidative stress.

Optimized labeling of bone marrow mesenchymal cells with superparamagnetic iron oxide nanoparticles and in vivo visualization by magnetic resonance imaging

PubMed Central

2011-01-01

Background Stem cell therapy has emerged as a promising addition to traditional treatments for a number of diseases. However, harnessing the therapeutic potential of stem cells requires an understanding of their fate in vivo. Non-invasive cell tracking can provide knowledge about mechanisms responsible for functional improvement of host tissue. Superparamagnetic iron oxide nanoparticles (SPIONs) have been used to label and visualize various cell types with magnetic resonance imaging (MRI). In this study we performed experiments designed to investigate the biological properties, including proliferation, viability and differentiation capacity of mesenchymal cells (MSCs) labeled with clinically approved SPIONs. Results Rat and mouse MSCs were isolated, cultured, and incubated with dextran-covered SPIONs (ferumoxide) alone or with poly-L-lysine (PLL) or protamine chlorhydrate for 4 or 24 hrs. Labeling efficiency was evaluated by dextran immunocytochemistry and MRI. Cell proliferation and viability were evaluated in vitro with Ki67 immunocytochemistry and live/dead assays. Ferumoxide-labeled MSCs could be induced to differentiate to adipocytes, osteocytes and chondrocytes. We analyzed ferumoxide retention in MSCs with or without mitomycin C pretreatment. Approximately 95% MSCs were labeled when incubated with ferumoxide for 4 or 24 hrs in the presence of PLL or protamine, whereas labeling of MSCs incubated with ferumoxide alone was poor. Proliferative capacity was maintained in MSCs incubated with ferumoxide and PLL for 4 hrs, however, after 24 hrs it was reduced. MSCs incubated with ferumoxide and protamine were efficiently visualized by MRI; they maintained proliferation and viability for up to 7 days and remained competent to differentiate. After 21 days MSCs pretreated with mitomycin C still showed a large number of ferumoxide-labeled cells. Conclusions The efficient and long lasting uptake and retention of SPIONs by MSCs using a protocol employing ferumoxide and protamine may be applicable to patients, since both ferumoxides and protamine are approved for human use. PMID:21542946

Effects of aerobic and anaerobic biological processes on leaching of heavy metals from soil amended with sewage sludge compost.

PubMed

Fang, Wen; Wei, Yonghong; Liu, Jianguo; Kosson, David S; van der Sloot, Hans A; Zhang, Peng

2016-12-01

The risk from leaching of heavy metals is a major factor hindering land application of sewage sludge compost (SSC). Understanding the change in heavy metal leaching resulting from soil biological processes provides important information for assessing long-term behavior of heavy metals in the compost amended soil. In this paper, 180days aerobic incubation and 240days anaerobic incubation were conducted to investigate the effects of the aerobic and anaerobic biological processes on heavy metal leaching from soil amended with SSC, combined with chemical speciation modeling. Results showed that leaching concentrations of heavy metals at natural pH were similar before and after biological process. However, the major processes controlling heavy metals were influenced by the decrease of DOC with organic matter mineralization during biological processes. Mineralization of organic matter lowered the contribution of DOC-complexation to Ni and Zn leaching. Besides, the reducing condition produced by biological processes, particularly by the anaerobic biological process, resulted in the loss of sorption sites for As on Fe hydroxide, which increased the potential risk of As release at alkaline pH. Copyright © 2016 Elsevier Ltd. All rights reserved.

Mechanical stiffness of TMJ condylar cartilage increases after artificial aging by ribose.

PubMed

Mirahmadi, Fereshteh; Koolstra, Jan Harm; Lobbezoo, Frank; van Lenthe, G Harry; Ghazanfari, Samaneh; Snabel, Jessica; Stoop, Reinout; Everts, Vincent

2018-03-01

Aging is accompanied by a series of changes in mature tissues that influence their properties and functions. Collagen, as one of the main extracellular components of cartilage, becomes highly crosslinked during aging. In this study, the aim was to examine whether a correlation exists between collagen crosslinking induced by artificial aging and mechanical properties of the temporomandibular joint (TMJ) condyle. To evaluate this hypothesis, collagen crosslinks were induced using ribose incubation. Porcine TMJ condyles were incubated for 7 days with different concentrations of ribose. The compressive modulus and stiffness ratio (incubated versus control) was determined after loading. Glycosaminoglycan and collagen content, and the number of crosslinks were analyzed. Tissue structure was visualized by microscopy using different staining methods. Concomitant with an increasing concentration of ribose, an increase of collagen crosslinks was found. The number of crosslinks increased almost 50 fold after incubation with the highest concentration of ribose. Simultaneously, the stiffness ratio of the samples showed a significant increase after incubation with the ribose. Pearson correlation analyses showed a significant positive correlation between the overall stiffness ratio and the crosslink level; the higher the number of crosslinks the higher the stiffness. The present model, in which ribose was used to mimic certain aspects of age-related changes, can be employed as an in vitro model to study age-related mechanical changes in the TMJ condyle. Copyright © 2017 Elsevier Ltd. All rights reserved.

Airborne concentrations of volatile organic compounds in neonatal incubators.

PubMed

Prazad, P; Cortes, D R; Puppala, B L; Donovan, R; Kumar, S; Gulati, A

2008-08-01

To identify and quantify airborne volatile organic compounds (VOCs) inside neonatal incubators during various modes of operation within the neonatal intensive care unit (NICU) environment. Air samples were taken from 10 unoccupied incubators in four operational settings along with ambient air samples using air sampling canisters. The samples were analyzed following EPA TO-15 using a Tekmar AutoCan interfaced to Agilent 6890 Gas Chromatograph with a 5973 Mass Spectrometer calibrated for 60 EPA TO-15 method target compounds. Non-target compounds were tentatively identified using mass spectral interpretation and with a mass spectral library created by National Institute for Standards and Technology. Two non-target compounds, 2-heptanone and n-butyl acetate, were found at elevated concentrations inside the incubators compared with ambient room air samples. Increase in temperature and addition of humidity produced further increased concentrations of these compounds. Their identities were verified by mass spectra and relative retention times using authentic standards. They were quantified using vinyl acetate and 2-hexanone as surrogate standards. The emission pattern of these two compounds and background measurements indicate that they originate inside the incubator. There is evidence that exposure to some VOCs may adversely impact the fetal and developing infants' health. Currently, as there is no definitive information available on the effects of acute or chronic low-level exposure to these compounds in neonates, future studies evaluating the health effects of neonatal exposure to these VOCs are needed.

Vasorelaxation responses to insulin in laminar vessel rings from healthy, lean horses.

PubMed

Wooldridge, A A; Waguespack, R W; Schwartz, D D; Venugopal, C S; Eades, S C; Beadle, R E

2014-10-01

Hyperinsulinemia causes laminitis experimentally and is a risk factor for naturally occurring laminitis. The aim of this study was to investigate the effects of insulin on laminar vascular relaxation and to induce insulin-associated vascular dysfunction in vitro. Relaxation responses of isolated laminar arterial and venous rings to acetylcholine and insulin were evaluated. To alter vascular function in response to insulin, all vessel rings were incubated with insulin or vehicle, submaximally contracted, administered insulin again and relaxation responses recorded. Laminar arteries were also incubated with the mitogen-activated protein kinase (MAPK) inhibitor, PD-98059. Relaxation in response to acetylcholine was not different between arteries and veins, but veins relaxed less in response to insulin than arteries. In arteries incubated with insulin, the subsequent relaxation response to insulin was blunted. Veins had minimal relaxation to insulin regardless of incubation. Arteries incubated with PD-98059 relaxed more in response to insulin than arteries not exposed to PD-98059, indicating that MAPK plays a role in maintenance of basal tone in laminar arteries. A differing response of laminar veins and arteries to insulin-induced relaxation may be important in understanding the link between hyperinsulinemia and laminitis. In vitro induction of vascular dysfunction in response to insulin in laminar arteries may be useful for testing therapeutic interventions and for understanding the pathophysiology of laminitis. Copyright © 2014 Elsevier Ltd. All rights reserved.

Gas chromatography/principal component similarity system for detection of E. coli and S. aureus contaminating salmon and hamburger.

PubMed

Nakai, S; Wang, Z H; Dou, J; Nakamura, S; Ogawa, M; Nakai, E; Vanderstoep, J

1999-02-01

Coho, Atlantic, Spring, and Sockeye salmon and five commercial samples of hamburger patties were analyzed by processing gas chromatography (GC) data of volatile compounds using the principal component similarity (PCS) technique. PCS scattergrams of the samples inoculated with Escherichia coli and Staphylococcus aureus followed by incubation showed the pattern-shift lines moving away from the data point for uninoculated, unincubated reference samples in different directions with increasing incubation time. When the PCS scattergrams were drawn for samples incubated overnight, the samples inoculated with the two bacterial species and the uninoculated samples appeared as three separated groups. This GC/PCS approach has the potential to ensure quality of samples by discriminating good samples from potentially spoiled samples. The latter may require further microbial assays to identify the bacteria species potentially contaminating foods.

Automation of metabolic stability studies in microsomes, cytosol and plasma using a 215 Gilson liquid handler.

PubMed

Linget, J M; du Vignaud, P

1999-05-01

A 215 Gilson liquid handler was used to automate enzymatic incubations using microsomes, cytosol and plasma. The design of automated protocols are described. They were based on the use of 96 deep well plates and on HPLC-based methods for assaying the substrate. The assessment of those protocols was made with comparison between manual and automated incubations, reliability and reproducibility of automated incubations in microsomes and cytosol. Examples of the use of those programs in metabolic studies in drug research, i.e. metabolic screening in microsomes and plasma were shown. Even rapid processes (with disappearance half lives as low as 1 min) can be analysed. This work demonstrates how stability studies can be automated to save time, render experiments involving human biological media less hazardous and may be improve inter-laboratory reproducibility.

Disaster incubation, cumulative impacts and the urban/ex-urban/rural dynamic

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mulvihill, Peter R.; Ali, S. Harris

2007-05-15

This article explores environmental impacts and risks that can accumulate in rural and ex-urban areas and regions and their relation to urban and global development forces. Two Southern Ontario cases are examined: an area level water disaster and cumulative change at the regional level. The role of disaster incubation analysis and advanced environmental assessment tools are discussed in terms of their potential to contribute to more enlightened and effective assessment and planning processes. It is concluded that conventional approaches to EA and planning are characteristically deficient in addressing the full range of impacts and risks, and particularly those originating frommore » pathogens, dispersed and insidious sources. Rigorous application of disaster incubation analysis and more advanced forms of EA has considerable potential to influence a different pattern of planning and decision making.« less

Methodological Considerations for Hair Cortisol Measurements in Children

PubMed Central

Slominski, Radomir; Rovnaghi, Cynthia R.; Anand, Kanwaljeet J. S.

2015-01-01

Background Hair cortisol levels are used increasingly as a measure for chronic stress in young children. We propose modifications to the current methods used for hair cortisol analysis to more accurately determine reference ranges for hair cortisol across different populations and age groups. Methods The authors compared standard (finely cutting hair) vs. milled methods for hair processing (n=16), developed a 4-step extraction process for hair protein and cortisol (n=16), and compared liquid chromatography-mass spectrometry (LCMS) vs. ELISA assays for measuring hair cortisol (n=28). The extraction process included sequential incubations in methanol and acetone, repeated twice. Hair protein was measured via spectrophotometric ratios at 260/280 nm to indicate the hair dissolution state using a BioTek® plate reader and dedicated software. Hair cortisol was measured using an ELISA assay kit. Individual (n=13), pooled hair samples (n=12) with high, intermediate, and low cortisol values and the ELISA assay internal standards (n=3) were also evaluated by LCMS. Results Milled and standard methods showed highly correlated hair cortisol (rs=0.951, p<0.0001) and protein values (rs=0.902, p=0.0002), although higher yields of cortisol and protein were obtained from the standard method in 13/16 and 14/16 samples respectively (p<0.05). Four sequential extractions yielded additional amounts of protein (36.5%, 27.5%, 30.5%, 3.1%) and cortisol (45.4%, 31.1%, 15.1%, 0.04%) from hair samples. Cortisol values measured by LCMS and ELISA were correlated (rs=0.737; p<0.0001), although cortisol levels (median [IQR]) detected in the same samples by LCMS (38.7 [14.4, 136] ng/ml) were lower than by ELISA (172.2 [67.9, 1051] ng/ml). LCMS also detected cortisone, which comprised 13.4% (3.7%, 25.9%) of the steroids detected. Conclusion Methodological studies suggest that finely cutting hair with sequential incubations in methanol and acetone, repeated twice, extracts greater yields of cortisol than does milled hair. Based on these findings, at least three incubations may be required to extract most of the cortisol in human hair samples. In addition, ELISA-based assays showed greater sensitivity for measuring hair cortisol levels than LCMS-based assays. PMID:25811341

Milk Thistle Extract and Silymarin Inhibit Lipopolysaccharide Induced Lamellar Separation of Hoof Explants in Vitro

PubMed Central

Reisinger, Nicole; Schaumberger, Simone; Nagl, Veronika; Hessenberger, Sabine; Schatzmayr, Gerd

2014-01-01

The pathogenesis of laminitis is not completely identified and the role of endotoxins (lipopolysaccharides, LPS) in this process remains unclear. Phytogenic substances, like milk thistle (MT) and silymarin, are known for their anti-inflammatory and antioxidant properties and might therefore have the potential to counteract endotoxin induced effects on the hoof lamellar tissue. The aim of our study was to investigate the influence of endotoxins on lamellar tissue integrity and to test if MT and silymarin are capable of inhibiting LPS-induced effects in an in vitro/ex vivo model. In preliminary tests, LPS neutralization efficiency of these phytogenics was determined in an in vitro neutralization assay. Furthermore, tissue explants gained from hooves of slaughter horses were tested for lamellar separation after incubation with different concentrations of LPS. By combined incubation of explants with LPS and either Polymyxin B (PMB; positive control), MT or silymarin, the influence of these substances on LPS-induced effects was assessed. In the in vitro neutralization assay, MT and silymarin reduced LPS concentrations by 64% and 75%, respectively, in comparison PMB reduced 98% of the LPS concentration. In hoof explants, LPS led to a concentration dependent separation. Accordantly, separation force was significantly decreased by 10 µg/mL LPS. PMB, MT and silymarin could significantly improve tissue integrity of explants incubated with 10 µg/mL LPS. This study showed that LPS had a negative influence on the structure of hoof explants in vitro. MT and silymarin reduced endotoxin activity and inhibited LPS-induced effects on the lamellar tissue. Hence, MT and silymarin might be used to support the prevention of laminitis and should be further evaluated for this application. PMID:25290524

Deferasirox is a powerful NF-κB inhibitor in myelodysplastic cells and in leukemia cell lines acting independently from cell iron deprivation by chelation and reactive oxygen species scavenging

PubMed Central

Messa, Emanuela; Carturan, Sonia; Maffè, Chiara; Pautasso, Marisa; Bracco, Enrico; Roetto, Antonella; Messa, Francesca; Arruga, Francesca; Defilippi, Ilaria; Rosso, Valentina; Zanone, Chiara; Rotolo, Antonia; Greco, Elisabetta; Pellegrino, Rosa M.; Alberti, Daniele; Saglio, Giuseppe; Cilloni, Daniela

2010-01-01

Background Usefulness of iron chelation therapy in myelodysplastic patients is still under debate but many authors suggest its possible role in improving survival of low-risk myelodysplastic patients. Several reports have described an unexpected effect of iron chelators, such as an improvement in hemoglobin levels, in patients affected by myelodysplastic syndromes. Furthermore, the novel chelator deferasirox induces a similar improvement more rapidly. Nuclear factor-κB is a key regulator of many cellular processes and its impaired activity has been described in different myeloid malignancies including myelodysplastic syndromes. Design and Methods We evaluated deferasirox activity on nuclear factor-κB in myelodysplastic syndromes as a possible mechanism involved in hemoglobin improvement during in vivo treatment. Forty peripheral blood samples collected from myelodysplastic syndrome patients were incubated with 50 μM deferasirox for 18h. Results Nuclear factor-κB activity dramatically decreased in samples showing high basal activity as well as in cell lines, whereas no similar behavior was observed with other iron chelators despite a similar reduction in reactive oxygen species levels. Additionally, ferric hydroxyquinoline incubation did not decrease deferasirox activity in K562 cells suggesting the mechanism of action of the drug is independent from cell iron deprivation by chelation. Finally, incubation with both etoposide and deferasirox induced an increase in K562 apoptotic rate. Conclusions Nuclear factor-κB inhibition by deferasirox is not seen from other chelators and is iron and reactive oxygen species scavenging independent. This could explain the hemoglobin improvement after in vivo treatment, such that our hypothesis needs to be validated in further prospective studies. PMID:20534700

Paradigm Diagnostics Salmonella Indicator Broth (PDX-SIB) for detection of Salmonella on selected environmental surfaces.

PubMed

Olstein, Alan; Griffith, Leena; Feirtag, Joellen; Pearson, Nicole

2013-01-01

The Paradigm Diagnostics Salmonella Indicator Broth (PDX-SIB) is intended as a single-step selective enrichment indicator broth to be used as a simple screening test for the presence of Salmonella spp. in environmental samples. This method permits the end user to avoid multistep sample processing to identify presumptively positive samples, as exemplified by standard U.S. reference methods. PDX-SIB permits the outgrowth of Salmonella while inhibiting the growth of competitive Gram-negative and -positive microflora. Growth of Salmonella-positive cultures results in a visual color change of the medium from purple to yellow when the sample is grown at 37 +/- 1 degree C. Performance of PDX-SIB has been evaluated in five different categories: inclusivity-exclusivity, methods comparison, ruggedness, lot-to-lot variability, and shelf stability. The inclusivity panel included 100 different Salmonella serovars, 98 of which were SIB-positive during the 30 to 48 h incubation period. The exclusivity panel included 33 different non-Salmonella microorganisms, 31 of which were SIB-negative during the incubation period. Methods comparison studies included four different surfaces: S. Newport on plastic, S. Anatum on sealed concrete, S. Abaetetuba on ceramic tile, and S. Typhimurium in the presence of 1 log excess of Citrobacter freundii. Results of the methods comparison studies demonstrated no statistical difference between the SIB method and the U.S. Food and Drug Administration-Bacteriological Analytical Manual reference method, as measured by the Mantel-Haenszel Chi-square test. Ruggedness studies demonstrated little variation in test results when SIB incubation temperatures were varied over a 34-40 degrees C range. Lot-to-lot consistency results suggest no detectable differences in manufactured goods using two reference Salmonella serovars and one non-Salmonella microorganism.

Importation of beef from countries infected with foot and mouth disease: a review of risk mitigation measures.

PubMed

Sutmoller, P

2001-12-01

Risk mitigation measures to reduce the risks associated with importing beef from countries affected by foot and mouth disease (FMD) consist of controls at the farm of origin, inspection of slaughterhouses and maturation and deboning of carcasses. This assessment evaluates the effect of these measures on the mitigation of the risks presented by meat from cattle with FMD, for each of the different stages of the disease. The four disease stages considered are the incubation period, the period of clinical signs, convalescence and the carrier stage. Efficient animal health systems, disease surveillance, and ante-mortem and post-mortem inspection of all cattle effectively reduce the risk of FMD transmission from cattle slaughtered during the period of clinical signs or convalescence. These measures fail if the cattle are slaughtered during the incubation period, because of the absence of clinical signs. Cattle in this stage of the infection are likely to be viraemic, with FMD virus present in the skeletal muscles. Maturation of the carcasses of viraemic cattle reduces the risk of virus presence in the beef. In addition, deboning and removal of the principal lymph nodes and large blood vessels eliminate a source of FMD contamination of the beef. However, the slaughter of viraemic cattle creates an additional hazard of gross environmental viral contamination of the slaughterhouse facilities. Therefore, the maturation process may create a false sense of security, and the emphasis should instead be placed on disease surveillance within the infected zone and on the farms of origin, to prevent the slaughter of herds that are incubating FMD. Cattle slaughtered during the carrier stage do not pose a risk for the international beef trade.

Fabrication of biogenic antimicrobial silver nanoparticles by Streptomyces aegyptia NEAE 102 as eco-friendly nanofactory.

PubMed

El-Naggar, Noura El-Ahmady; Abdelwahed, Nayera A M; Darwesh, Osama M M

2014-04-01

The current research was focused on the extracellular biosynthesis of bactericidal silver nanoparticles (AgNPs) using cell-free supernatant of a local isolate previously identified as a novel Streptomyces aegyptia NEAE 102. The biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102 was quite fast and required far less time than previously published strains. The produced particles showed a single surface plasmon resonance peak at 400 nm by UV-Vis spectroscopy, which confirmed the presence of AgNPs. Response surface methodology was chosen to evaluate the effects of four process variables (AgNO3 concentration, incubation period, pH levels, and inoculum size) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. Statistical analysis of the results showed that the linear and quadratic effects of incubation period, initial pH, and inoculum size had a significant effect (p < 0.05) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. The maximum silver nanoparticles biosynthesis (2.5 OD, at 400 nm ) was achieved in runs number 5 and 14 under the conditions of 1 mM AgNO3 (1-1.5% (v/v)), incubation period (72-96 h), initial pH (9-10), and inoculum size (2-4% (v/v)). An overall 4-fold increase in AgNPs biosynthesis was obtained as compared with that of unoptimized conditions. The biosynthesized silver nanoparticles were characterized using UV-VIS spectrophotometer and Fourier transform infrared spectroscopy analysis, in addition to antimicrobial properties. The biosynthesized AgNPs significantly inhibited the growth of medically important pathogenic gram-positive (Staphylococcus aureus) and gram-negative bacteria (Pseudomonas aeruginosa) and yeast (Candida albicans).

Milk thistle extract and silymarin inhibit lipopolysaccharide induced lamellar separation of hoof explants in vitro.

PubMed

Reisinger, Nicole; Schaumberger, Simone; Nagl, Veronika; Hessenberger, Sabine; Schatzmayr, Gerd

2014-10-06

The pathogenesis of laminitis is not completely identified and the role of endotoxins (lipopolysaccharides, LPS) in this process remains unclear. Phytogenic substances, like milk thistle (MT) and silymarin, are known for their anti-inflammatory and antioxidant properties and might therefore have the potential to counteract endotoxin induced effects on the hoof lamellar tissue. The aim of our study was to investigate the influence of endotoxins on lamellar tissue integrity and to test if MT and silymarin are capable of inhibiting LPS-induced effects in an in vitro/ex vivo model. In preliminary tests, LPS neutralization efficiency of these phytogenics was determined in an in vitro neutralization assay. Furthermore, tissue explants gained from hooves of slaughter horses were tested for lamellar separation after incubation with different concentrations of LPS. By combined incubation of explants with LPS and either Polymyxin B (PMB; positive control), MT or silymarin, the influence of these substances on LPS-induced effects was assessed. In the in vitro neutralization assay, MT and silymarin reduced LPS concentrations by 64% and 75%, respectively, in comparison PMB reduced 98% of the LPS concentration. In hoof explants, LPS led to a concentration dependent separation. Accordantly, separation force was significantly decreased by 10 µg/mL LPS. PMB, MT and silymarin could significantly improve tissue integrity of explants incubated with 10 µg/mL LPS. This study showed that LPS had a negative influence on the structure of hoof explants in vitro. MT and silymarin reduced endotoxin activity and inhibited LPS-induced effects on the lamellar tissue. Hence, MT and silymarin might be used to support the prevention of laminitis and should be further evaluated for this application.

Influence of serotonin on the action of melatonin in MIH-induced meiotic resumption in the oocytes of carp Catla catla.

PubMed

Chattoraj, Asamanja; Seth, Mohua; Maitra, Saumen Kumar

2008-07-01

The influences of serotonin (5-hydroxytryptamine) on the action of melatonin (N-acetyl-5-methoxytryptamine) in MIH (maturation inducing hormone)-induced meiotic resumption were evaluated in the oocytes of carp Catla catla using an in vitro model. Oocytes from gravid female carp were isolated and incubated separately in Medium 199 containing either (a) only melatonin (MEL; 100 pg/mL), or (b) only serotonin (SER; 100 pg/mL), or (c) only MIH (1 microg/mL), or (d) MEL and MIH (e) or MEL (4 h before) and MIH, or (f) MEL and SER, (g) or SER and MIH, or (h) SER (4 h before) and MIH, or (i) luzindole (L-antagonist of MEL receptors; 10 microM) and MEL, or (j) MEL, L and MIH, or (k) MEL (4 h before), L and MIH, or (l) metoclopramide hydrochloride (M-antagonist of SER receptors; 10 microM) and SER, or (m) M, MEL, SER, or (n) M, SER and MIH, or (o) M, SER (4 h before) and MIH, or (p) M, MEL SER and MIH, or (q) MEL, L, SER and M, or (r) MEL, L, SER, M, and MIH, or (s) MEL, SER, L and MIH. Control oocytes were incubated in the medium alone. Oocytes were incubated for 4, or 8, or 12, or 16 h and effects were evaluated by considering the rate (%) of germinal vesicle breakdown (GVBD). At the end of 16 h incubation, 93.24+/-1.57% oocytes underwent GVBD following incubation with only MIH, while incubation with only MEL or only SER resulted in 77.15+/-1.91% or 14.42+/-0.43% GVBD respectively. Interestingly, incubation with MEL 4 h prior to addition of MIH in the medium, led to an accelerated rate of GVBD (92.58+/-1.10% at 12 h). In contrast, SER, irrespective of its time of application in relation to MIH, resulted in a maximum of 64.57+/-0.86% GVBD. While L was found to reduce the stimulatory actions of melatonin, M suppressed the inhibitory actions of serotonin. In each case, both electrophoretic and immunoblot studies revealed that the rate of GVBD was associated with the rate of formation of maturation promoting factor (a complex of two proteins: a regulatory component--cyclin B and the catalytic component--Cdk1 or cdc2). Collectively, the present study reports for the first time that SER not only inhibits the independent actions of MIH, but also the actions of MEL on the MIH-induced oocytes maturation in carp.

Evaluation of the effect of process variables on the fatty acid profile of single cell oil produced by Mortierella using solid-state fermentation.

PubMed

Asadi, Seyedeh Zeinab; Khosravi-Darani, Kianoush; Nikoopour, Houshang; Bakhoda, Hossein

2015-03-01

This article reviews some of the aspects of single cell oil (SCO) production using solid-state fermentation (SSF) by fungi of the genus Mortierella. This article provides an overview of the advantages of SSF for SCO formation by the aforementioned fungus and demonstrates that the content of the polyunsaturated fatty acids (PUFA) depend on the type of fermentation media and culture conditions. Process variables that influence lipid accumulation by Mortierella spp. and the profile of the fatty acids are discussed, including incubation temperature, time, aeration, growth phase of the mycelium, particle size of the substrate, carbon to nitrogen ratio, initial moisture content and pH as well as supplementation of the substrate with nitrogen and oil. Finally, the article highlights future research trends for the scaled-up production of PUFAs in SSF.

Enteropathogenic Escherichia coli Outbreak and its Incubation Period: Is it Short or Long?

PubMed

Lee, Dong-Woo; Gwack, Jin; Youn, Seun-Ki

2012-03-01

The aim of this study is to determine the incubation period of enteropathogenic Escherichia coli (EPEC), which creates several outbreaks in a year in South Korea. We reviewed all water and food-borne outbreaks data reported to the Korea Centers for Disease Control and Prevention (KCDC) from 2009 to 2010 and determined their characteristics. Through this process, we can presume the incubation period of EPEC among outbreaks in South Korea. A total of 497 water and food-borne outbreaks were reported to KCDC and 66 (13.28%) are defined as E coli-origin outbreaks. EPEC was the most common subtype of E coli, being confirmed as a causative organism in 26 outbreaks. Overall attack rate was 15.85% (range 0.9-100). The subjects were eight outbreaks that have a clear history of single exposure and we can estimate the incubation time of EPEC as minimum 0.5 hours to maximum 34.0 hours with a mean 12.9 hours (range 4.5-24.0). The cases of those cannot completely rule out the chance of multiple exposure from same source or place have minimum 1.0 hour, to a maximum of 195.5 hours and a mean 30.5 (range 22.7-61.0) hours of incubation period. This serial analysis suggests that EPEC has actually shorter mean incubation period as much as 12 hours. When this period is longer than 1 day or over, then the epidemiologic investigator should consider the chance of repeated or continuous exposure by making it clear whether there is any chance of any other exposure in common.

Calcein AM release-based cytotoxic cell assay for fish leucocytes.

PubMed

Iwanowicz, Luke R; Densmore, Christine L; Ottinger, Christopher A

2004-02-01

A non-specific cytotoxic cell assay for fish is presented that is based on the release of the activated fluorochrome calcein AM from lysed carp epithelioma papulosum cyprini (EPC) cells. To establish the suitability of treating EPC cells with calcein AM the uptake and spontaneous release of the calcein AM by the EPC cells was evaluated. Incubation of 5 microM calcein AM in culture medium with 1x10(5)EPC cells well(-1)for a minimum of 3 h provided sufficient labelling. Spontaneous release of fluorescence from the labelled EPC cells during 10 h of post labelling incubation ranged from 30 to 39% of the total observed fluorescence. Cytotoxic activity of trout leucocytes was evaluated at three leucocyte to target cell ratios (10:1, 2:1 and 1:1) following incubation (4, 6, 8, and 10 h) with calcein AM-labelled EPC cells at 15 degrees C. In some instances, the monoclonal antibody specific for the NCC surface receptor NCCRP-1 (MAb5C.6) was included in the cultures. The activity of NCC cells was significantly inhibited in the presence of 0.25 microg well(-1)of MAb5C.6 relative to no antibody (P

The responses of immune cells to iron oxide nanoparticles.

PubMed

Xu, Yaolin; Sherwood, Jennifer A; Lackey, Kimberly H; Qin, Ying; Bao, Yuping

2016-04-01

Immune cells play an important role in recognizing and removing foreign objects, such as nanoparticles. Among various parameters, surface coatings of nanoparticles are the first contact with biological system, which critically affect nanoparticle interactions. Here, surface coating effects on nanoparticle cellular uptake, toxicity and ability to trigger immune response were evaluated on a human monocyte cell line using iron oxide nanoparticles. The cells were treated with nanoparticles of three types of coatings (negatively charged polyacrylic acid, positively charged polyethylenimine and neutral polyethylene glycol). The cells were treated at various nanoparticle concentrations (5, 10, 20, 30, 50 μg ml(-1) or 2, 4, 8, 12, 20 μg cm(-2)) with 6 h incubation or treated at a nanoparticle concentration of 50 μg ml(-1) (20 μg cm(-2)) at different incubation times (6, 12, 24, 48 or 72 h). Cell viability over 80% was observed for all nanoparticle treatment experiments, regardless of surface coatings, nanoparticle concentrations and incubation times. The much lower cell viability for cells treated with free ligands (e.g. ~10% for polyethylenimine) suggested that the surface coatings were tightly attached to the nanoparticle surfaces. The immune responses of cells to nanoparticles were evaluated by quantifying the expression of toll-like receptor 2 and tumor necrosis factor-α. The expression of tumor necrosis factor-α and toll-like receptor 2 were not significant in any case of the surface coatings, nanoparticle concentrations and incubation times. These results provide useful information to select nanoparticle surface coatings for biological and biomedical applications. Copyright © 2016 John Wiley & Sons, Ltd.

On the crystallization of amorphous germanium films

NASA Astrophysics Data System (ADS)

Edelman, F.; Komem, Y.; Bendayan, M.; Beserman, R.

1993-06-01

The incubation time for crystallization of amorphous Ge (a-Ge) films, deposited by e-gun, was studied as a function of temperature between 150 and 500°C by means of both in situ transmission electron microscopy and Raman scattering spectroscopy. The temperature dependence of t0 follows an Arrhenius curve with an activation energy of 2.0 eV for free-sustained a-Ge films. In the case where the a-Ge films were on Si 3N 4 substrate, the activation energy of the incubation process was 1.3 eV.

Oxygen and nitrogen isotope effects duing nitrification and denitrification occuring in Midwesern soils

NASA Astrophysics Data System (ADS)

Michalski, G. M.; Wilkens, B.; Sanchez, A. V.; Yount, J.

2017-12-01

The processes of nitrification and denitrification are key steps in the biogeochemical cycling of N and are a main control on ecosystem productivity. These processes are ephemeral and often difficult to assess across wide spatial and temporal scales. Natural abundance stable isotopes are a way of potentially assessing these two processes across multiple scales. We have conducted incubation experiments to assess the N and O isotope effects occurring during denitrification in soils typical of the Midwestern United States. Nitrification was examined by incubating soils amended with ammonium (with a known δ15N) mixed with H2O and O2 that had different δ18O values and then measured the δ15N and δ18O of the product nitrate. The fraction of nitrate oxygen arising from H2O and O2 was determined along with the N and O kinetic isotope effect (KIE). For denitrification, nitrate with a known δ15N, δ17O, and δ18O, was incubated in anaerobic soils from 12-48 hours. The residual nitrate was analyzed for isotope change and the KIE for O and N as well as exchange with H2O was determined. These data can be useful for interpreting nitrate isotopes in agricultural fields as a way off assessing nitrification and denitrification is agricultural ecosystems such as the IML-CZO.

Decolorization and Degradation of Batik Dye Effluent using Ganoderma lucidum

NASA Astrophysics Data System (ADS)

Pratiwi, Diah; Indrianingsih, A. W.; Darsih, Cici; Hernawan

2017-12-01

Batik is product of traditional Indonesia culture that developed into a large textile industry. Synthetic dyes which widely used in textile industries including batik. Colour can be removed from wastewater effluent by chemical, physical, and biology methods. Bioremediation is one of the methods that used for processing colored effluent. Isolated White-rot fungi Ganoderma lucidum was used for bioremediation process for batik effluent. G. lucidum was developed by G. lucidum cultivation on centers of mushroom farmer Media Agro Merapi Kaliurang, Yogyakarta. The batik effluent was collected from a private small and medium Batik enterprises located at Petir, Rongkop, Gunungkidul Regency. The aim of the study were to optimize decolorization of Naphtol Black (NB) using G. lucidum. The effect of process parameters like incubation time and dye concentration on dye decolorization and COD degradation was studied. G. lucidum were growth at pH 5-6 and temperature 25°C at various Naphtol Black dye with concentration 20 ppm, 50 ppm, and 100 ppm for 30 day incubation time. The result from this study increased decolorization in line with the increasing of COD degradation. Increasing percentage of decolorization and COD degradation gradually increased with incubation time and dye concentration. The maximum decolorization and COD reduction were found to be 60,53% and 81,03%. G. lucidum had potential to decolorized and degraded COD for NB dye effluent on higher concentration.

Photodegradation of fresh terrigenous organic matter contributes to the heterotrophy of boreal aquatic ecosystems in Quebec (Canada)

NASA Astrophysics Data System (ADS)

Plouhinec, J.; Lucotte, M. M.; Ouellet, A.; Gelinas, Y.

2012-12-01

The processes that fuel heterotrophy and thus contribute to CO2 production in lakes and reservoirs of the boreal region in Quebec are still not fully understood. To shed light on some of the factors controlling heterotrophy, we evaluated the importance of photodechemical mineralization of dissolved organic mater relative to other sources of CO2 production in six natural or human-perturbed lakes through logging on their watersheds and two reservoirs of the Quebec boreal forest over a period of 1.5 year. Rates of CO2 production in the water column were measured through incubation/irradiation experiments, using a series of filtrations to isolate the effects of photochemical mineralization, bacterial respiration, and planktonic respiration. Total CO2 fluxes measured in this study compared well to total diffusive fluxes measured through the traditional thin boundary layer method, thus validating our incubation approach. We calculated the daily integrated production of CO2 through photochemical mineralization (DIPMCO2) of dissolved organic matter over the entire water column using the calculation of the spectrum yield (Φλ). DIPMCO2 appeared as a robust indicator strongly correlated to the absorption coefficient of chromophoric dissolved organic matter (CDOM) at 360 nm (R2=0.81, p<0.01). DIPMCO2 accounts for 15% ± 14% of the total diffusive flux of CO2 to the atmosphere, independently of water body type or perturbation level. Our data also suggests that photochemical mineralization and photosynthesis processes are strongly correlated (R2=0.79, p<0.01), which is due to the fact that the strong photosynthetically active radiation (PAR) attenuation derives from a terrestrial organic matter (TOM) input into the water column. Also, the total diffusive fluxes of CO2 towards the atmosphere (fCO2) are correlated to the DIPMCO2 values (R2=0.49, p<0.01). We have evaluated theoretical CO2 fluxes emitted from the photic zone (f°CO2 ) by photochemical mineralization, bacterial respiration and CO2 consumption by phosynthesis based on measurements obtained from incubation/irradiation experiments. Finally, the relationship between fCO2 and the theoretical CO2 fluxes after production and consumption of CO2 considered only in the epilimnitic zone (R2=0.97, p<0.01) shows that, independently of the environmental conditions, the passive CO2 fluxes are approximately equal to 10 mmol.CO2.m-2.d-1.

Effects of incubator temperature and oxygen concentration during the plateau stage of oxygen consumption on Turkey embryo long bone development.

PubMed

Oviedo-Rondón, E O; Small, J; Wineland, M J; Christensen, V L; Grimes, J L; Funderburk, S V L; Ort, D T; Mann, K M

2008-08-01

Temperature (TEM) and O(2) concentrations during the plateau stage of oxygen consumption are known to affect yolk utilization, tissue development, and thyroid metabolism in turkey embryos. Three experiments were conducted to evaluate these incubation effects on long bone development. Fertile eggs of Nicholas turkeys were used. In each trial, standard incubation conditions were used to 24 d, when the eggs containing viable embryos were randomly divided into 4 groups. Four experimental cabinets provided 4 TEM (36, 37, 38, or 39 degrees C) or 4 O(2) concentrations (17, 19, 21, or 23% O(2)). In the third experiment, 2 temperatures (36 and 39 degrees C) and 2 O(2) concentrations (17 and 23%) were evaluated in a 2 x 2 factorial design. Body and residual yolk weights were obtained. Both legs were dissected, and shanks, femur, and tibia weights, length, and thickness were recorded. Relative asymmetry of each leg section was calculated. Chondrocyte density was evaluated in slides stained with hematoxylin and eosin. Immunofluorescence was used to evaluate the presence of collagen type X and transforming growth factor beta. Hot TEM caused reduction of tibia weights and increase of shank weight when compared with cool TEM. The lengths of femur, tibia, and shanks were reduced by 39 degrees C. The relative asymmetry of leg weights were increased at 38 and 39 degrees C. Poult body and part weights were not affected by O(2) concentrations, but poults on 23% O(2) had bigger shanks and heavier tibias than the ones on 17% O(2). High TEM depressed the fluorescence of collagen type X and transforming growth factor beta. The O(2) concentrations did not consistently affect the immunofluorescence of these proteins. The chondrocyte density was affected by TEM and O(2) in resting and hypertrophic zones. In the third experiment, high TEM depressed BW, leg muscle weights, and shank length. Low O(2) reduced tibia and shanks as a proportion of the whole body. We concluded that incubation conditions affect long bone development in turkeys.

Formation of biofilm by Listeria monocytogenes ATCC 19112 at different incubation temperatures and concentrations of sodium chloride

PubMed Central

Lee, H.Y.; Chai, L.C.; Pui, C.F.; Mustafa, S.; Cheah, Y.K.; Nishibuchi, M.; Radu, S.

2013-01-01

Biofilm formation can lead to various consequences in the food processing line such as contamination and equipment breakdowns. Since formation of biofilm can occur in various conditions; this study was carried out using L. monocytogenes ATCC 19112 and its biofilm formation ability tested under various concentrations of sodium chloride and temperatures. Cultures of L. monocytogenes ATCC 19112 were placed in 96-well microtitre plate containing concentration of sodium chloride from 1–10% (w/v) and incubated at different temperature of 4 °C, 30 °C and 45 °C for up to 60 h. Absorbance reading of crystal violet staining showed the density of biofilm formed in the 96-well microtitre plates was significantly higher when incubated in 4 °C. The formation of biofilm also occurs at a faster rate at 4 °C and higher optical density (OD 570 nm) was observed at 45 °C. This shows that storage under formation of biofilm that may lead to a higher contamination along the processing line in the food industry. Formation of biofilm was found to be more dependent on temperature compared to sodium chloride stress. PMID:24159283

Changes in hepatic glucose and lipid metabolism-related parameters in domestic pigeon (Columba livia) during incubation and chick rearing.

PubMed

Wan, X P; Xie, P; Bu, Z; Zou, X T

2018-04-01

This study aimed to evaluate the hepatic glucose and lipid metabolism-related parameters of adult male and female White King pigeons (Columba livia) during incubation and chick rearing. At day 4 (I4), 10 (I10) and 17 (I17) of incubation and day 1 (R1), 7 (R7), 15 (R15) and 25 (R25) of chick rearing, livers were sampled from six pigeons for each sex. Glycogen and fat contents, activities of glycolytic enzymes (hexokinase, HK; 6-phosphofructokinase, 6-PFK), and genes expressions of key enzymes involved in glycolysis (pyruvate kinase, PK; glucokinase, GK), gluconeogenesis (phosphoenolpyruvate carboxykinase cytosolic, PCK1; fructose-1,6-bisphosphatase, FBP1; glucose-6-phosphatase, G6Pase), fatty acid synthesis (fatty acid synthase, FAS; acetyl-CoA carboxylase, ACC) and fatty acid β-oxidation (carnitine palmitoyltransferase 1, CPT1; acyl-CoA 1, ACO) were measured. In male and female pigeon livers, glycogen content and HK activity dramatically increased after I17 and after R1, respectively; expressions of FBP1 and G6Pase genes were maximized at R15; activity of 6-PFK and expressions of PK and CPT1 genes were highest at R7; fat content and expressions of FAS and ACC genes steeply increased from I10 to R1. In females, hepatic expressions of GK and PCK1 genes were greatest at R7 and I17, respectively; however, in males, both of them were maximized at R15. Hepatic expression of ACO gene was significantly enhanced at R1 compared to I17 and R7 in males, whereas it was notably up-regulated at I17 and R7 in females. Furthermore, expressions of PCK1, GK, FAS and ACC genes were in significant relation to fat content in the livers of female pigeons, while fat content in male pigeons was highly correlated with expression of PCK1, ACC, CPT1 and ACO genes. In conclusion, regulations of glucose and lipid metabolic processes were enhanced in parent pigeon livers from terminal phases of incubation to mid phase of chick rearing with sexual effects. © 2017 Blackwell Verlag GmbH.

21 CFR 640.91 - Processing.

Code of Federal Regulations, 2012 CFR

2012-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma Protein Fraction (Human) § 640.91 Processing. (a) Date... °C or colder. (e) Heat treatment. Heating of the final containers of Plasma Protein Fraction (Human... concentration of the product. (g) Incubation. All final containers of Plasma Protein Fraction (Human) shall be...

21 CFR 640.91 - Processing.

Code of Federal Regulations, 2011 CFR

2011-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma Protein Fraction (Human) § 640.91 Processing. (a) Date... °C or colder. (e) Heat treatment. Heating of the final containers of Plasma Protein Fraction (Human... concentration of the product. (g) Incubation. All final containers of Plasma Protein Fraction (Human) shall be...

21 CFR 640.91 - Processing.

Code of Federal Regulations, 2013 CFR

2013-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma Protein Fraction (Human) § 640.91 Processing. (a) Date... °C or colder. (e) Heat treatment. Heating of the final containers of Plasma Protein Fraction (Human... concentration of the product. (g) Incubation. All final containers of Plasma Protein Fraction (Human) shall be...

21 CFR 640.91 - Processing.

Code of Federal Regulations, 2014 CFR

2014-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma Protein Fraction (Human) § 640.91 Processing. (a) Date... °C or colder. (e) Heat treatment. Heating of the final containers of Plasma Protein Fraction (Human... concentration of the product. (g) Incubation. All final containers of Plasma Protein Fraction (Human) shall be...

Triiodothyronine stimulates cartilage growth and maturation by different mechanisms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Burch, W.M.; Van Wyk, J.J.

1987-02-01

The mechanisms by which triiodothyronine (T3) stimulates growth and maturation of growth-plate cartilage in vitro were studied by incubating embryonic chick pelvic cartilages in serum-free medium in the presence and absence of T3 for 3 days. To determine whether T3 might stimulate production of somatomedins by the cartilage, medium from cartilage incubated with and without T3 was assayed for somatomedin C( Sm-C) by radioimmunoassay. No difference in Sm-C content was found. However, cartilage incubated with T3 and increasing amounts of human Sm-C (0.5-20 ng/ml) weighed more and had greater amounts of glycosaminoglycan that cartilage incubated in the same concentrations ofmore » Sm-C without T3, suggesting that T3 enhances the growth effect of somatomedin. The authors added a monoclonal antibody to Sm-C (anti-Sm-C) to the organ culture to determine whether T3's stimulatory effect on cartilage growth could be blocked. The anti-Sm-C inhibited growth of cartilage incubated in medium alone and blocked the growth response to T3. They propose two different mechanisms by which T3 affects growth-plate cartilage: (1) T3 promotes cartilage growth primarily through enhancing the effect of somatomedin, and (2) T3 stimulates cartilage maturation possibly by accelerating the normal process of cartilage differentiation from proliferative to hypertrophic chondrocytes.« less

A combined study of heat and mass transfer in an infant incubator with an overhead screen.

PubMed

Ginalski, Maciej K; Nowak, Andrzej J; Wrobel, Luiz C

2007-06-01

The main objective of this study is to investigate the major physical processes taking place inside an infant incubator, before and after modifications have been made to its interior chamber. The modification involves the addition of an overhead screen to decrease radiation heat losses from the infant placed inside the incubator. The present study investigates the effect of these modifications on the convective heat flux from the infant's body to the surrounding environment inside the incubator. A combined analysis of airflow and heat transfer due to conduction, convection, radiation and evaporation has been performed, in order to calculate the temperature and velocity fields inside the incubator before and after the design modification. Due to the geometrical complexity of the model, computer-aided design (CAD) applications were used to generate a computer-based model. All numerical calculations have been performed using the commercial computational fluid dynamics (CFD) package FLUENT, together with in-house routines used for managing purposes and user-defined functions (UDFs) which extend the basic solver capabilities. Numerical calculations have been performed for three different air inlet temperatures: 32, 34 and 36 degrees C. The study shows a decrease of the radiative and convective heat losses when the overhead screen is present. The results obtained were numerically verified as well as compared with results available in the literature from investigations of dry heat losses from infant manikins.

Dynamics of two methanogenic microbiomes incubated in polycyclic aromatic hydrocarbons, naphthenic acids, and oil field produced water.

PubMed

Oko, Bonahis J; Tao, Yu; Stuckey, David C

2017-01-01

Oil field produced water (OFPW) is widely produced in large volumes around the world. Transforming the organic matter in OFPW into bioenergy, such as biomethane, is one promising way to sustainability. However, OFPW is difficult to biologically degrade because it contains complex compounds such as naphthenic acids (NAs), or polycyclic aromatic hydrocarbons (PAHs). Although active microbial communities have been found in many oil reservoirs, little is known about how an exotic microbiome, e.g. the one which originates from municipal wastewater treatment plants, would evolve when incubated with OFPW. In this study, we harvested methanogenic biomass from two sources: a full-scale anaerobic digester (AD) treating oil and gas processing wastewater (named O&G sludge), and from a full-scale AD reactor treating multiple fractions of municipal solid wastes (named MS, short for mixed sludge). Both were incubated in replicate microcosms fed with PAHs, NAs, or OFPW. The results showed that the PAHs, NAs, and OFPW feeds could rapidly alter the methanogenic microbiomes, even after 14 days, while the O&G sludge adapted faster than the mixed sludge in all the incubations. Two rarely reported microorganisms, a hydrogenotrophic methanogen Candidatus methanoregula and a saccharolytic fermenter Kosmotoga , were found to be prevalent in the PAHs and OFPW microcosms, and are likely to play an important role in the syntrophic degradation of PAHs and OFPW, cooperating with methanogens such as Methanoregula, Methanosarcina, or Methanobacterium . The dominant phyla varied in certain patterns during the incubations, depending on the biomass source, feed type, and variation in nutrients. The sludge that originated from the oil and gas processing wastewater treatment (O&G) reactor adapted faster than the one from municipal solid waste reactors, almost certainly because the O&G biomass had been "pre-selected" by the environment. This study reveals the importance of biomass selection for other crude oil-waste-related bioengineering studies, such as bioaugmentation and bioremediation.

Deriving fractional rate of degradation of logistic-exponential (LE) model to evaluate early in vitro fermentation.

PubMed

Wang, M; Sun, X Z; Tang, S X; Tan, Z L; Pacheco, D

2013-06-01

Water-soluble components of feedstuffs are mainly utilized during the early phase of microbial fermentation, which could be deemed an important determinant of gas production behavior in vitro. Many studies proposed that the fractional rate of degradation (FRD) estimated by fitting gas production curves to mathematical models might be used to characterize the early incubation for in vitro systems. In this study, the mathematical concept of FRD was developed on the basis of the Logistic-Exponential (LE) model, with initial gas volume being zero (LE0). The FRD of the LE0 model exhibits a continuous increase from initial (FRD 0) toward final asymptotic value (FRD F) with longer incubation time. The relationships between the FRD and gas production at incubation times 2, 4, 6, 8, 12 and 24 h were compared for four models, in addition to LE0, Generalization of the Mitscherlich (GM), c th order Michaelis-Menten (MM) and Exponential with a discrete LAG (EXPLAG). A total of 94 in vitro gas curves from four subsets with a wide range of feedstuffs from different laboratories and incubation periods were used for model testing. Results indicated that compared with the GM, MM and EXPLAG models, the FRD of LE0 model consistently had stronger correlations with gas production across the four subsets, especially at incubation times 2, 4, 6, 8 and 12 h. Thus, the LE0 model was deemed to provide a better representation of the early fermentation rates. Furthermore, the FRD 0 also exhibited strong correlations (P < 0.05) with gas production at early incubation times 2, 4, 6 and 8 h across all four subsets. In summary, the FRD of LE0 model provides an alternative to quantify the rate of early stage incubation, and its initial value could be an important starting parameter of rate.

Characterizing early embryonic development of Brown Tsaiya Ducks (Anas platyrhynchos) in comparison with Taiwan Country Chicken (Gallus gallus domestics)

PubMed Central

Lumsangkul, Chompunut; Fan, Yang-Kwang; Chang, Shen-Chang; Ju, Jyh-Cherng

2018-01-01

Avian embryos are among the most convenient and the primary representatives for the study of classical embryology. It is well-known that the hatching time of duck embryos is approximately one week longer than that of chicken embryos. However, the key features associated with the slower embryonic development in ducks have not been adequately described. This study aimed to characterize the pattern and the speed of early embryogenesis in Brown Tsaiya Ducks (BTD) compared with those in Taiwan Country Chicken (TCC) by using growth parameters including embryonic crown-tail length (ECTL), primitive streak formation, somitogenesis, and other development-related parameters, during the first 72 h of incubation. Three hundred and sixty eggs from BTD and TCC, respectively, were incubated at 37.2°C, and were then dissected hourly to evaluate their developmental stages. We found that morphological changes of TCC embryos shared a major similarity with that of the Hamburger and Hamilton staging system during early chick embryogenesis. The initial primitive streak in TCC emerged between 6 and 7 h post-incubation, but its emergence was delayed until 10 to 13 h post-incubation in BTD. Similarly, the limb primordia (wing and limb buds) were observed at 51 h post-incubation in TCC embryos compared to 64 h post-incubation in BTD embryos. The allantois first appeared around 65 to 68 h in TCC embryos, but it was not observed in BTD embryos. At the 72 h post-incubation, 40 somites were clearly formed in TCC embryos while only 32 somites in BTD embryos. Overall, the BTD embryos developed approximately 16 h slower than the chicken embryo during the first 72 h of development. To our best knowledge, this is the first study to describe two distinct developmental time courses between TCC and BTD, which would facilitate future embryogenesis-related studies of the two important avian species in Taiwan. PMID:29742160

Evaluation of 100 brain examinations using a 3 Tesla MR-compatible incubator-safety, handling, and image quality.

PubMed

Sirin, Selma; Goericke, Sophia L; Huening, Britta M; Stein, Anja; Kinner, Sonja; Felderhoff-Mueser, Ursula; Schweiger, Bernd

2013-10-01

Several studies have revealed the importance of brain imaging in term and preterm infants. The aim of this retrospective study was to review safety, handling, and image quality of MR brain imaging using a new 3 Tesla MR-compatible incubator. Between 02/2011 and 05/2012 100 brain MRIs (84 infants, mean gestational age 32.2 ± 4.7 weeks, mean postmenstrual age at imaging 40.6 ± 3.4 weeks) were performed using a 3 Tesla MR-compatible incubator with dedicated, compatible head coil. Seventeen examinations (13 infants, mean gestational age 35.1 ± 5.4 weeks, mean postmenstrual age at imaging 47.8 ± 7.4 weeks) with a standard head coil served as a control. Image analysis was performed by a neuroradiologist and a pediatric radiologist in consensus. All but two patients with known apnea were transferred to the MR unit and scanned without problems. Handling was easier and faster with the incubator; relevant motion artifacts (5.9 vs. 10.8%) and the need for repetitive sedation (43.0 vs. 86.7%) were reduced. Considering only images not impaired by motion artifacts, image quality (4.8 ± 0.4 vs. 4.3 ± 0.8, p = 0.047) and spatial resolution (4.7 ± 0.4 vs. 4.2 ± 0.6, p = 0.011) of T2-weighted images were scored significantly higher in patients imaged with the incubator. SNR increased significantly (171.6 ± 54.5 vs. 80.5 ± 19.8, p < 0.001) with the use of the incubator. Infants can benefit from the use of a 3 Tesla MR-compatible incubator because of its safety, easier, and faster handling (compared to standard imaging) and possibility to obtain high-quality MR images even in unstable patients.

Validation Study of Rapid Assays of Bioburden, Endotoxins and Other Contamination.

PubMed

Shintani, Hideharu

2016-01-01

Microbial testing performed in support of pharmaceutical and biopharmaceutical production falls into three main categories: detection (qualitative), enumeration (quantitative), and characterization/identification. Traditional microbiological methods are listed in the compendia and discussed by using the conventional growth-based techniques, which are labor intensive and time consuming. In general, such tests require several days of incubation for microbial contamination (bioburden) to be detected, and therefore management seldom is able to take proactive corrective measures. In addition, microbial growth is limited by the growth medium used and incubation conditions, thus impacting testing sensitivity, accuracy, and reproducibility. 　For more than 20 years various technology platforms for rapid microbiological methods (RMM) have been developed, and many have been readily adopted by the food industry and clinical microbiology laboratories. Their use would certainly offer drug companies faster test turnaround times to accommodate the aggressive deadlines for manufacturing processes and product release. Some rapid methods also offer the possibility for real-time microbial analyses, enabling management to respond to microbial contamination events in a more timely fashion, and can provide cost savings and higher efficiencies in quality control testing laboratories. Despite the many proven business and quality benefits and the fact that the FDA's initiative to promote the use of process analytical technology (PAT) includes rapid microbial methods, pharmaceutical and biopharmaceutical industries have been somewhat slow to embrace alternative microbial methodologies for several reasons. The major reason is that the bioburden counts detected by the incubation method and rapid assay are greatly divergent. 　The use of rapid methods is a dynamic field in applied microbiology and one that has gained increased attention nationally and internationally over time. This topic has been extensively addressed at conferences and in published documents around the world. More recently, the use of alternative methods for control of the microbiological quality of pharmaceutical products and materials used in pharmaceutical production has been addressed by the compendia in an attempt to facilitate implementation of these technologies by pharmaceutical companies. The author presents some of the rapid method technologies under evaluation or in use by pharmaceutical microbiologists and the current status of the implementation of alternative microbial methods.

Melatonin accelerates maturation inducing hormone (MIH): induced oocyte maturation in carps.

PubMed

Chattoraj, Asamanja; Bhattacharyya, Sharmistha; Basu, Dipanjan; Bhattacharya, Shelley; Bhattacharya, Samir; Maitra, Saumen Kumar

2005-02-01

The present communication is an attempt to demonstrate the influence of melatonin on the action of maturation inducing hormone (MIH) on the maturation of oocytes in carps. The oocytes from gravid female major carp Labeo rohita were isolated and incubated separately in Medium 199 containing (a) only MIH (1 microg/ml), (b) only melatonin (at concentrations of 50, 100 or 500 pg/ml), and (c) both melatonin and MIH, but at different time intervals. In the latter group, melatonin was added to the incubating medium either (i) 4 h before addition of MIH, (ii) 2 h before addition of MIH, (iii) co-administered with MIH (0 h interval) or (iv) 2 h after addition of MIH. In each case, oocytes were further incubated for 4, 8, 12 or 16 h post- administration of MIH, and the effects of treatment on oocyte maturation were evaluated by considering the rate (%) of germinal vesicle breakdown (GVBD). Incubation of oocytes in a medium containing only melatonin did not result in GVBD of any oocyte. Nearly all the oocytes underwent GVBD when incubated with MIH for 16 h. Administration of melatonin along with MIH (at 0 h interval) or 2 h after addition of MIH did not result in any significant change in the rate of GVBD compared to that in a medium containing only MIH. However, it was quite interesting to observe that incubation of oocytes with melatonin especially 4 h prior to addition of MIH in the medium, led to an accelerated rate of GVBD in the oocytes. Experiments with the oocytes of another major carp Cyprinus carpio following an identical schedule depicted similar results except a difference in the optimum melatonin dose. In L. rohita, 50 pg/ml melatonin had maximum acceleratory effect on MIH-induced GVBD of oocytes, while it was 100 pg/ml in C. carpio. Further study revealed that pre-incubation with melatonin accelerates the action of MIH on the formation of a complex of two proteins (MPF), a regulatory component called cyclin B and the catalytic component protein kinase known as cyclin-dependent kinase, Cdk1. Densitometric analysis of the immunoblot data collected from the melatonin pre-treated MIH incubated oocytes showed that cyclin B level continued to increase even after 4 h of incubation, and reached the peak after 12 h. Moreover, determination of H1 kinase activity as an indicator of MPF activity in oocytes revealed that melatonin pre-incubation considerably increased MIH stimulation of histone H1 phosphorylation as compared to MIH alone. Thus, the present study demonstrates for the first time that prior incubation with melatonin accelerates the action of MIH on carp oocyte maturation.

Evaluation of the bioNexia Legionella Test, Including Impact of Incubation Time Extension, for Detection of Legionella pneumophila Serogroup 1 Antigen in Urine.

PubMed

Badoux, Paul; Euser, Sjoerd M; Bruin, Jacob P; Mulder, Patrick P G; Yzerman, Ed P F

2017-06-01

In this study, we compared the bioNexia test (bioMérieux, Marcy-l'Étoile, France), a new immunochromatographic assay for the detection of Legionella pneumophila serogroup 1 in urine, with the BinaxNOW urinary antigen test (Alere, Waltham, Massachusetts, USA). After 15 min of incubation (in accordance with the manufacturers' instructions), the sensitivities and specificities were, respectively, 76.5% and 97.2% for the bioNexia test and 87.1% and 100% for the BinaxNOW test. After a prolonged incubation time of 60 min, the sensitivities and specificities increased to, respectively, 89.4% and 97.2% for the bioNexia test and 91.8% and 100% for the BinaxNOW test. When the tests were read after 15 min, the concentration of discrepant urine samples increased the sensitivities to 94.1% for both tests. In conclusion, we found that although the bioNexia test showed lower sensitivity for the detection of L. pneumophila antigen in nonconcentrated urine compared to the BinaxNOW test, a prolonged incubation time as well as the use of concentrated samples showed comparable sensitivities for both tests. Copyright © 2017 American Society for Microbiology.

In vitro studies of chicken egg yolk antibody (IgY) against Salmonella enteritidis and Salmonella typhimurium.

PubMed

Lee, E N; Sunwoo, H H; Menninen, K; Sim, J S

2002-05-01

Chicken egg yolk antibody (IgY) raised against Salmonella enteritidis or Salmonella typhimurium was found in highly specific activity levels by ELISA. S. enteritidis- and S. typhimurium-specific IgY powder, prepared by freeze-drying the egg yolk water-soluble fraction, contained 15.5 and 10.0% of specific IgY, respectively. Anti-S. enteritidis IgY cross-reacted 55.3% with S. typhimurium. The cross-reactivity of anti-S. typhimurium IgY with S. enteritidis was 42.4%. Salmonella-specific IgY was demonstrated to inhibit Salmonella growth in liquid medium. The growth rate of S. enteritidis incubated with S. enteritidis-specific IgY was fourfold less than that of the control group during a 4-to-6-h incubation. Cell counts of S. typhimurium incubated with S. typhimurium-specific IgY were reduced by 1.6 log cfu/mL in comparison to that of the control group after 6 h of incubation. The specific binding activity of IgY was further evaluated by using immunofluorescence and immunoelectron microscopy. It was found that Salmonella-specific IgY could bind to the antigens expressed on the Salmonella surface, resulting in structural alterations of the bacterial surface.

Rate of penicillin killing of Staphylococcus aureus and Autobac 1 susceptibility test results.

PubMed Central

Harris, J A; Furtado, D

1982-01-01

A clinical isolate of Staphylococcus aureus interpreted as resistant to penicillin by the Autobac 1 susceptibility testing method (i.e., light-scattering index of 0.77) was found to be susceptible to penicillin by both the disk diffusion and broth dilution techniques. The growth rate of the clinical isolate during a 4-h incubation interval was similar to that of a known sensitive reference strain (S. aureus ATCC 25923) used as a control organism for the Autobac test. The bactericidal effect of penicillin was evaluated by measuring the rate of killing over a 4-h interval. The percentages of organisms surviving exposure to 5.0 or 2.5 U of penicillin G per ml (number of organisms recovered at 3 h/number of organisms introduced as inoculum) were 68 and 76%, respectively, for the clinical isolate and 15 and 21%, respectively, for the reference strain. After 24 h of incubation, penicillin was bactericidal for both strains. The need to increase the time of incubation for those S. aureus isolates resistant to penicillin after 3 h of standard incubation time in the Autobac system is discussed. PMID:7068821

A cost for high levels of sperm competition in rodents: increased sperm DNA fragmentation

PubMed Central

Roldan, Eduardo R. S.

2016-01-01

Sperm competition, a prevalent evolutionary process in which the spermatozoa of two or more males compete for the fertilization of the same ovum, leads to morphological and physiological adaptations, including increases in energetic metabolism that may serve to propel sperm faster but that may have negative effects on DNA integrity. Sperm DNA damage is associated with reduced rates of fertilization, embryo and fetal loss, offspring mortality, and mutations leading to genetic disease. We tested whether high levels of sperm competition affect sperm DNA integrity. We evaluated sperm DNA integrity in 18 species of rodents that differ in their levels of sperm competition using the sperm chromatin structure assay. DNA integrity was assessed upon sperm collection, in response to incubation under capacitating or non-capacitating conditions, and after exposure to physical and chemical stressors. Sperm DNA was very resistant to physical and chemical stressors, whereas incubation in non-capacitating and capacitating conditions resulted in only a small increase in sperm DNA damage. Importantly, levels of sperm competition were positively associated with sperm DNA fragmentation across rodent species. This is the first evidence showing that high levels of sperm competition lead to an important cost in the form of increased sperm DNA damage. PMID:26936246

Adhesion and invasion of Clostridium perfringens type A into epithelial cells.

PubMed

Llanco, Luis A; Nakano, Viviane; Moraes, Claudia T P de; Piazza, Roxane M F; Avila-Campos, Mario J

Clostridium perfringens is the causative agent for necrotic enteritis. It secretes the major virulence factors, and α- and NetB-toxins that are responsible for intestinal lesions. The TpeL toxin affects cell morphology by producing myonecrosis, but its role in the pathogenesis of necrotic enteritis is unclear. In this study, the presence of netB and tpeL genes in C. perfringens type A strains isolated from chickens with necrotic enteritis, their cytotoxic effects and role in adhesion and invasion of epithelial cells were evaluated. Six (27.3%) of the 22 C. perfringens type A strains were harboring the tpeL gene and produced morphological alterations in Vero cells after 6h of incubation. Strains tpeL (-) induced strong cell rounding after 6h of incubation and produced cell enlargement. None of the 22 strains harbored netB gene. All the six tpeL (+) gene strains were able to adhere to HEp-2 cells; however, only four of them (66.6%) were invasive. Thus, these results suggest that the presence of tpeL gene or TpeL toxin might be required for the adherence of bacteria to HEp-2 cells; however, it could not have any role in the invasion process. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Bioremediation of PAH-contamined soils: Consequences on formation and degradation of polar-polycyclic aromatic compounds and microbial community abundance.

PubMed

Biache, Coralie; Ouali, Salma; Cébron, Aurélie; Lorgeoux, Catherine; Colombano, Stéfan; Faure, Pierre

2017-05-05

A bioslurry batch experiment was carried out over five months on three polycyclic aromatic compound (PAC) contaminated soils to study the PAC (PAH and polar-PAC) behavior during soil incubation and to evaluate the impact of PAC contamination on the abundance of microbial communities and functional PAH-degrading populations. Organic matter characteristics and reactivity, assessed through solvent extractable organic matter and PAC contents, and soil organic matter mineralization were monitored during 5 months. Total bacteria and fungi, and PAH-ring hydroxylating dioxygenase genes were quantified. Results showed that PAHs and polar-PACs were degraded with different degradation dynamics. Differences in degradation rates were observed among the three soils depending on PAH distribution and availability. Overall, low molecular weight compounds were preferentially degraded. Degradation selectivity between isomers and structurally similar compounds was observed which could be used to check the efficiency of bioremediation processes. Bacterial communities were dominant over fungi and were most likely responsible for PAC degradation. Abundance of PAH-degrading bacteria increased during incubations, but their proportion in the bacterial communities tended to decrease. The accumulation of some oxygenated-PACs during the bioslurry experiment underlines the necessity to monitor these compounds during application of remediation treatment on PAH contaminated soils. Copyright © 2017 Elsevier B.V. All rights reserved.

The inhibitory potential of the condensed-tannin-rich fraction of Plathymenia reticulata Benth. (Fabaceae) against Bothrops atrox envenomation.

PubMed

de Moura, Valéria Mourão; da Silva, Wania Cristina Rodrigues; Raposo, Juliana D A; Freitas-de-Sousa, Luciana A; Dos-Santos, Maria Cristina; de Oliveira, Ricardo Bezerra; Veras Mourão, Rosa Helena

2016-05-13

Ethnobotanical studies have shown that Plathymenia reticulata Benth. (Fabaceae) has been widely used in cases of snake envenomation, particularly in Northern Brazil. In light of this, the aim of this study was to evaluate the inhibitory potential of the condensed-tannin-rich fraction obtained from the bark of P. reticulata against the main biological activities induced by Bothrops atrox venom (BaV). The chemical composition of the aqueous extract of P. reticulata (AEPr) was first investigated by thin-layer chromatography (TLC) and the extract was then fractionated by column chromatography on Sephadex LH-20. This yielded five main fractions (Pr1, Pr2, Pr3, Pr4 and Pr5), which were analyzed by colorimetry to determine their concentrations of total phenolics, total tannins and condensed tannins and to assess their potential for blocking the phospholipase activity of BaV. The Pr5 fraction was defined as the fraction rich in condensed tannins (CTPr), and its inhibitory potential against the activities of the venom was evaluated. CTPr was evaluated in different in vivo and in vitro experimental protocols. The in vivo protocols consisted of (1) pre-incubation (venom:CTPr, w/w), (2) pre-treatment (orally administered) and (3) post-treatment (orally administered) to evaluate the effect on the hemorrhagic and edematogenic activities of BaV; in the in vitro protocol the effect on phospholipase and coagulant activity using pre-incubation in both tests was evaluated. There was statistically significant inhibition (p<0.05) of hemorrhagic activity by CTPr when the pre-incubation protocol was used [55% (1:5, w/w) and 74% (1:10, w/w)] and when pre-treatment with doses of 50 and 100mg/kg was used (19% and 13%, respectively). However, for the concentrations tested, there was no statistically significant inhibition in the group subjected to post-treatment administered orally. CTPr blocked 100% of phospholipase activity and 63.3% (1:10, w/w) of coagulant activity when it was pre-incubated with BaV. There was a statistically significant reduction (p<0.05) in edema induced by BaV in the oral protocols. Maximum inhibition was 95% (pre-treatment). Our findings indicate that CTPr could be a good source of natural inhibitors of the components of snake venom responsible for inducing local inflammation. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Elasticity and breaking strength of synthetic suture materials incubated in various equine physiological and pathological solutions.

PubMed

Kearney, C M; Buckley, C T; Jenner, F; Moissonnier, P; Brama, P A J

2014-07-01

Selection of suture material in equine surgery is often based on costs or subjective factors, such as the surgeon's personal experience, rather than objective facts. The amount of objective data available on durability of suture materials with regard to specific equine physiological conditions is limited. To evaluate the effect of various equine physiological and pathological fluids on the rate of degradation of a number of commonly used suture materials. In vitro material testing. Suture materials were exposed in vitro to physiological fluid, followed by biomechanical analysis. Three absorbable suture materials, glycolide/lactide copolymer, polyglactin 910 and polydioxanone were incubated at 37°C for 7, 14 or 28 days in phosphate-buffered saline, equine serum, equine urine and equine peritoneal fluid from an animal with peritonitis. Five strands of each suture material type were tested to failure in a materials testing machine for each time point and each incubation medium. Yield strength, strain and Young's modulus were calculated, analysed and reported. For all suture types, the incubation time had a significant effect on yield strength, percentage elongation and Young's modulus in all culture media (P<0.0001). Suture type was also shown significantly to influence changes in each of yield strength, percentage elongation and Young's modulus in all culture media (P<0.0001). While the glycolide/lactide copolymer demonstrated the highest Day 0 yield strength, it showed the most rapid degradation in all culture media. For each of the 3 material characteristics tested, polydioxanone showed the least variation across the incubation period in each culture medium. The duration of incubation and the type of fluid have significant effects on the biomechanical properties of various suture materials. These findings are important for evidence-based selection of suture material in clinical cases. © 2013 EVJ Ltd.

Knot Security of 5 Metric (USP 2) Sutures: Influence of Knotting Technique, Suture Material, and Incubation Time for 14 and 28 Days in Phosphate Buffered Saline and Inflamed Equine Peritoneal Fluid.

PubMed

Sanders, Ruth E; Kearney, Clodagh M; Buckley, Conor T; Jenner, Florien; Brama, Pieter A

2015-08-01

To evaluate knot security for 3 knot types created in 3 commonly used 5 metric suture materials incubated in physiological and pathological fluids. In vitro mechanical study. Knotted suture loops (n = 5/group). Loops of 3 different suture materials (glycolide/lactide copolymer; polyglactin 910; polydioxanone) were created around a 20 mm rod using 3 knot types (square [SQ], surgeon's [SK], and triple knot [TK]) and were tested to failure in distraction (6 mm/min) after tying (day 0) and after being incubated for 14 and 28 days in phosphate buffered saline (PBS) or inflamed peritoneal fluid. Failure load (N) and mode were recorded and compared. For polydioxanone, significant differences in force to knot failure were found between SQ and SK/TK but not between SK and TK. The force required to break all constructs increased after incubation in phosphate buffered saline (PBS). With glycolide/lactide copolymer no differences in force to knot failure were observed. With polyglactin 910, a significant difference between SQ and TK was observed, which was not seen between the other knot types. Incubation in inflamed peritoneal fluid caused a larger and more rapid decrease in force required to cause knot failure than incubation in PBS. Mechanical properties of suture materials have significant effects on knot security. For polydioxanone, SQ is insufficient to create a secure knot. Additional wraps above a SK confer extra stability in some materials, but this increase may not be clinically relevant or justifiable. Glycolide/lactide copolymer had excellent knot security. © Copyright 2015 by The American College of Veterinary Surgeons.

Periodical low eggshell temperatures during incubation and post hatch dietary arginine supplementation: Effects on performance and cold tolerance acquisition in broilers.

PubMed

Afsarian, O; Shahir, M H; Akhlaghi, A; Lotfolahian, H; Hoseini, A; Lourens, A

2016-10-01

An experiment was conducted to evaluate the effects of a periodically low eggshell temperature exposure during incubation and dietary supplementation of arginine on performance, ascites incidence, and cold tolerance acquisition in broilers. A total of 2,400 hatching eggs were randomly assigned to 2 treatment groups (16 replicates of 75 eggs per treatment). The eggs were incubated at a constant eggshell temperature (EST) of 37.8ºC throughout the incubation period (CON) or were periodically exposed to 15°C for one hour on days 11, 13, 15, and 17 of incubation and the EST was measured (periodical low EST; PLE). After hatching, 240 one-day-old male broiler chicks from both treatment groups were reared for 42 d with or without dietary arginine supplementation in a completely randomized design with a 2 × 2 factorial arrangement. In order to induce ascites, all chicks were exposed to a 15°C room temperature from 14 d onwards. Results showed that second grade chicks and yolk sac weight were decreased, and final body weight was increased in the PLE group. Ascites mortality rate was decreased only in the PLE group and dietary arginine supplementation had no apparent effect. In the PLE group, the packed cell volume (PCV) percentage and red blood cell (RBC) count were decreased. In conclusion, the results showed that the PLE treatment during incubation was associated with improved hatchability, chick quality, and productive performance of broilers and decreased ascites incidence during post hatch cold exposure. Dietary arginine supplementation had no beneficial effects in cold exposed broilers. © 2016 Poultry Science Association Inc.

Information content of incubation experiments for inverse estimation of pools in the Rothamsted carbon model: a Bayesian perspective

NASA Astrophysics Data System (ADS)

Scharnagl, B.; Vrugt, J. A.; Vereecken, H.; Herbst, M.

2010-02-01

A major drawback of current soil organic carbon (SOC) models is that their conceptually defined pools do not necessarily correspond to measurable SOC fractions in real practice. This not only impairs our ability to rigorously evaluate SOC models but also makes it difficult to derive accurate initial states of the individual carbon pools. In this study, we tested the feasibility of inverse modelling for estimating pools in the Rothamsted carbon model (ROTHC) using mineralization rates observed during incubation experiments. This inverse approach may provide an alternative to existing SOC fractionation methods. To illustrate our approach, we used a time series of synthetically generated mineralization rates using the ROTHC model. We adopted a Bayesian approach using the recently developed DiffeRential Evolution Adaptive Metropolis (DREAM) algorithm to infer probability density functions of the various carbon pools at the start of incubation. The Kullback-Leibler divergence was used to quantify the information content of the mineralization rate data. Our results indicate that measured mineralization rates generally provided sufficient information to reliably estimate all carbon pools in the ROTHC model. The incubation time necessary to appropriately constrain all pools was about 900 days. The use of prior information on microbial biomass carbon significantly reduced the uncertainty of the initial carbon pools, decreasing the required incubation time to about 600 days. Simultaneous estimation of initial carbon pools and decomposition rate constants significantly increased the uncertainty of the carbon pools. This effect was most pronounced for the intermediate and slow pools. Altogether, our results demonstrate that it is particularly difficult to derive reasonable estimates of the humified organic matter pool and the inert organic matter pool from inverse modelling of mineralization rates observed during incubation experiments.

Oviductal epithelial cells selected boar sperm according to their functional characteristics

PubMed Central

López-Úbeda, Rebeca; García-Vázquez, Francisco A; Gadea, Joaquín; Matás, Carmen

2017-01-01

The interaction of oviductal epithelial cells (OECs) with the spermatozoa has beneficial effects on the sperm functions. The aim of this study is to evaluate the in vitro fertilizing capacity of incubating spermatozoa previously selected by density gradient in OEC and determinate some sperm characteristics that could explain the results obtained. In this study, we assessed in vitro fertilization (IVF), tyrosine phosphorylation, phosphatidylserine translocation, nuclear DNA fragmentation, and chromatin decondensation. Three experimental sperm groups, previously selected by Percoll gradient, were established according to the origin of the sperm used for IVF: (i) W30 group: spermatozoa were incubated with oocytes in the absence of OEC; (ii) NB group: after sperm incubation in OEC, the unbound spermatozoa were incubated with oocytes, in the absence of OEC; and (iii) B group: after sperm incubation with OEC, the bound spermatozoa were incubated with oocytes in the OEC plates. The results showed that sperm from the NB group led to a lower IVF yield, accompanied by low penetration rates (NB: 19.6%, B: 94.9%, and W30: 62.9%; P < 0.001) and problems of nuclear decondensation. Moreover, higher levels of tyrosine phosphorylation were observed in the NB group compared with the W30 and B groups (NB: 58.7%, B: 2.5%, and W30: 4.5%; P < 0.01). A similar trend was observed in phosphatidylserine translocation (NB: 93.7%, B: 5.7%, and W30: 44.2%; P < 0.01). These results demonstrate that the OEC exerts a rigorous degree of sperm selection, even within an already highly selected population of spermatozoa, and can capture the best functional spermatozoa for fertilization. PMID:27232850

Effects of fluoridated milk on root dentin remineralization.

PubMed

Arnold, Wolfgang H; Heidt, Bastian A; Kuntz, Sebastian; Naumova, Ella A

2014-01-01

The prevalence of root caries is increasing with greater life expectancy and number of retained teeth. Therefore, new preventive strategies should be developed to reduce the prevalence of root caries. The aim of this study was to investigate the effects of fluoridated milk on the remineralization of root dentin and to compare these effects to those of sodium fluoride (NaF) application without milk. Thirty extracted human molars were divided into 6 groups, and the root cementum was removed from each tooth. The dentin surface was demineralized and then incubated with one of the following six solutions: Sodium chloride NaCl, artificial saliva, milk, milk+2.5 ppm fluoride, milk+10 ppm fluoride and artificial saliva+10 ppm fluoride. Serial sections were cut through the lesions and investigated with polarized light microscopy and quantitative morphometry, scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). The data were statistically evaluated using a one-way ANOVA for multiple comparisons. The depth of the lesion decreased with increasing fluoride concentration and was the smallest after incubation with artificial saliva+10 ppm fluoride. SEM analysis revealed a clearly demarcated superficial remineralized zone after incubation with milk+2.5 ppm fluoride, milk+10 ppm fluoride and artificial saliva+10 ppm fluoride. Ca content in this zone increased with increasing fluoride content and was highest after artificial saliva+10 ppm fluoride incubation. In the artificial saliva+10 ppm fluoride group, an additional crystalline layer was present on top of the lesion that contained elevated levels of F and Ca. Incubation of root dentin with fluoridated milk showed a clear effect on root dentin remineralization, and incubation with NaF dissolved in artificial saliva demonstrated a stronger effect.

Is newborn melatonin production influenced by magnetic fields produced by incubators?

PubMed

Bellieni, Carlo Valerio; Tei, Monica; Iacoponi, Francesca; Tataranno, Maria Luisa; Negro, Simona; Proietti, Fabrizio; Longini, Mariangela; Perrone, Serafina; Buonocore, Giuseppe

2012-08-01

During permanence in most incubators, newborns are very close to the electric engine, which represents a source of electromagnetic fields (EMF). Previous studies demonstrated a decrease in melatonin production in adults and animals exposed to EMF. To assess melatonin production in a group of newborns exposed to EMF, and to evaluate whether removing the babies from the source of MF can affect melatonin production. We have recruited 28 babies (study group), who had spent at least 48 h in incubator where we had previously assessed the presence of significant EMF. We have measured their mean 6-hydroxy-melatonin-sulfate (6OHMS) urine excretion at the end of their permanence in the incubators, and compared it with their mean 6OHMS excretion after having been put in cribs, where EMF are below the detectable limit (<0.1mG). We have also measured urine 6OHMS twice, with an interval of 48h, in a control group of 27 babies who were not exposed to EMF during both samples. Mean 6OHMS/cr values were respectively 5.34±4.6 and 7.68±5.1ng/mg (p=0.026) when babies were exposed to EMF in incubators, and after having been put in the crib. In the control group, mean 6OHMS/cr values in the first and in the second sample were respectively 5.91±5.41 vs 6.17±3.94ng/mg (p=0.679). The transitory increase in melatonin production soon after removing newborns from incubators demonstrates a possible influence of EMF on melatonin production in newborns. Further studies are needed to confirm these data. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

Use of hydrogen peroxide during incubation of landlocked fall Chinook salmon eggs in vertical-flow incubators

USGS Publications Warehouse

Barnes, M.E.; Gaikowski, M.P.

2004-01-01

Six different hydrogen peroxide treatment regimes were evaluated in a series of three trials with landlocked fall Chinook salmon Oncorhynchus tshawytscha eggs incubated in vertical-flow incubators. Six daily 15-min hydrogen peroxide treatment regimes (1,000 mg/L; 1,000 mg/L with a decrease to 500 mg/L during estimated blastopore formation; 2,000 mg/L; 2,000 mg/L with a decrease to 500 mg/L during estimated blastopore formation; 2,500 mg/L; and 2,500 mg/L with a decrease to 500 mg/L during estimated blastopore formation) were compared with daily 15-min treatments of 1,667 mg/L of formalin. Mortality at egg eye-up and fry hatch and from eye-up to hatch was significantly greater in eggs receiving the 2,500-mg/L hydrogen peroxide treatments throughout incubation and in those receiving 2,500 mg/L hydrogen peroxide with a decrease to 500 mg/L during blastopore formation than in either of the 1,000-mg/L hydrogen peroxide treatment regimes or the formalin-treated eggs in the first trial. No significant differences in mortality were observed among any of the treatments in the subsequent two trials with maximum hydrogen peroxide concentrations of 2,000 mg/L. Fungal infestations were observed primarily in the incubation trays treated at either of the 1,000-mg/L hydrogen peroxide regimens, as well as in those trays whose treatment concentrations were dropped to 500 mg/L during blastopore formation. Infestations were not observed in any of the formalin-treated trays. If minor fungal infestation is acceptable, then daily hydrogen peroxide treatments of 1,000 mg/L for 15 min would probably provide adequate fungal control compared with formalin usage.

Effect of artificial saliva with different pH levels on the cytotoxicity of soft denture lining materials.

PubMed

Akay, Canan; Tanış, Merve Ç; Sevim, Handan

2017-10-13

The aim of this study was to evaluate the cytotoxic effects of 9 different soft denture liners on the viability of L-929 mouse fibroblast cells at different incubation periods by storing them in artificial saliva (AS) with different pH levels. 96 disk samples from each lining material were prepared and divided into 4 groups: GI: No treatment; GII: Stored in artificial saliva with pH 3 for 21 days; Group III: Stored in artificial saliva with pH 7 for 21 days; and Group IV: Stored in artificial saliva with pH 14 for 21 days. The cytotoxicity of the extracts to cultured mouse fibroblasts (L-929) was measured by MTT (tetrazolium salt 3-[4,5-dimethylthiazol-2-yl]-2,5-dipHnyltetrazolium bromide) assay. Data were analyzed using 1-way analysis of variation (ANOVA). It was found that for the pH 3 values of New Truliner, Trusoft, Mollosil Plus, Dentusil, TDV, and HydroCast®; for the pH 7 values of Ufi Gel P and Elite plus; and for the pH 14 values of HydroCast®, there was a noncytotoxic effect during both the 24-hour and 48-hour incubation periods. In the control group 48-hour incubation period, HydroCast®, TDV, Mollosil, 24-hour incubation period Elite plus, for pH 3 values; Elite Plus 24-hour incubation period, for pH 7 values Trusoft 48-hour incubation period there was a moderately cytotoxic effect. This in vitro study revealed that storage in artificial saliva with different pH levels can affect the cytotoxicity of soft lining materials.

Biomimetic evaluation of β tricalcium phosphate prepared by hot isostatic pressing

PubMed Central

Mateescu, Mihaela; Rguitti, Emmanuelle; Ponche, Arnaud; Descamps, Michel; Anselme, Karine

2012-01-01

Two types of completely densified β-TCP tablets were synthesized from a stoichiometric β-TCP powder. The first ones (TCP) were conventionally sintered, while the second ones (TCP-T) were sintered and treated by hot isostatic process (HIP). The HIP produced completely densified materials with relative densities greater than 99.9% and a transparent appearance of tablets. Samples were immersed in culture medium with (CM) or without serum (NCM) in static and dynamic conditions for a biomimetic evaluation. Similarly, SaOs-2 cells were cultured on samples in a static or dynamic flow perfusion system. The results of surface transformation in absence of cells showed that the dynamic condition increased the speed of calcium phosphate precipitations compared with the static condition. The morphology of precipitates was different with nature of tablets. The immersion in CM did impede this precipitation. XPS analysis of TCP-T tablets showed the presence of hydroxyapatite (HA) precipitates after incubation in NCM while octacalcium phosphate (OCP) precipitates were formed after incubation in CM. The analysis of the response of SaOs-2 cells on surfaces showed that the two types of materials are biocompatible. However, the dynamic mode of culture stimulated the differentiation of cells. Finally, it appears that the HIP treatment of TCP produces highly densified and transparent samples that display a good in vitro biocompatibility in static and dynamic culture conditions. Moreover, an interesting result of this work is the relationship between the presence of proteins in the immersion medium and the quality of precipitates formed on hipped TCP surface. PMID:23507861

Evaluation of UV-C mutagenized Scheffersomyces stipitis strains for ethanol production

USDA-ARS?s Scientific Manuscript database

We evaluated fermentation capabilities of five strains of Scheffersomyces stipitis (WT-2-1, WT-1-11, 14-2-6, 22-1-1, and 22-1-12) that had been produced by UV-C mutagenesis and selection for improved xylose fermentation to ethanol using an integrated automated robotic work cell. They were incubated ...

Leptin inhibits testosterone secretion from adult rat testis in vitro.

PubMed

Tena-Sempere, M; Pinilla, L; González, L C; Diéguez, C; Casanueva, F F; Aguilar, E

1999-05-01

Leptin, the product of the ob gene, has emerged recently as a pivotal signal in the regulation of fertility. Although the actions of leptin in the control of reproductive function are thought to be exerted mainly at the hypothalamic level, the potential direct effects of leptin at the pituitary and gonadal level have been poorly characterised. In the present study, we first assessed the ability of leptin to regulate testicular testosterone secretion in vitro. Secondly, we aimed to evaluate whether leptin can modulate basal gonadotrophin and prolactin (PRL) release by incubated hemi-pituitaries from fasted male rats. To attain the first goal, testicular slices from prepubertal and adult rats were incubated with increasing concentrations (10(-9)-10(-7) M) of recombinant leptin. Assuming that in vitro testicular responsiveness to leptin may be dependent on the background leptin levels, testicular tissue from both food-deprived and normally-fed animals was used. Furthermore, leptin modulation of stimulated testosterone secretion was evaluated by incubation of testicular samples with different doses of leptin in the presence of 10 IU human chorionic gonadotrophin (hCG). In addition, analysis of leptin actions on pituitary function was carried out using hemi-pituitaries from fasted adult male rats incubated in the presence of increasing concentrations (10(-9)-10(-7) M) of recombinant leptin. Serum testosterone levels, and basal and hCG-stimulated testosterone secretion by incubated testicular tissue were significantly decreased by fasting in prepubertal and adult male rats. However, a significant reduction in circulating LH levels was only evident in adult fasted rats. Doses of 10(-9)-10(-7) M leptin had no effect on basal or hCG-stimulated testosterone secretion by testes from prepubertal rats, regardless of the nutritional state of the donor animal. In contrast, leptin significantly decreased basal and hCG-induced testosterone secretion by testes from fasted and fed adult rats. In addition, 10(-9) M leptin inhibited LH and FSH secretion by incubated hemi-pituitaries from fasted adult males, whereas, at all doses tested, it was ineffective in modulating PRL release. Our results show that leptin, depending on the state of sexual maturation, is able to inhibit testosterone secretion acting at the testicular level. Furthermore, the present data suggest that the actions of leptin on the reproductive system are complex and are probably carried out at different levels of the hypothalamic-pituitary-gonadal axis.

Marketing Plan for Demonstration and Validation Assets

DOE Office of Scientific and Technical Information (OSTI.GOV)

None, None

The National Security Preparedness Project (NSPP), is to be sustained by various programs, including technology demonstration and evaluation (DEMVAL). This project assists companies in developing technologies under the National Security Technology Incubator program (NSTI) through demonstration and validation of technologies applicable to national security created by incubators and other sources. The NSPP also will support the creation of an integrated demonstration and validation environment. This report documents the DEMVAL marketing and visibility plan, which will focus on collecting information about, and expanding the visibility of, DEMVAL assets serving businesses with national security technology applications in southern New Mexico.

Evaluation of a New Chromogenic Medium (StrepB Select) for Detection of Group B Streptococcus from Vaginal-Rectal Specimens ▿

PubMed Central

Louie, L.; Kotowich, L.; Meaney, H.; Vearncombe, M.; Simor, A. E.

2010-01-01

We compared StrepB Select medium (Select) after enrichment with conventional culture for the detection of Group B Streptococcus (GBS). Postenrichment sensitivities of Select and conventional culture were 98.8% and 92.2%, respectively (P < 0.05). Select was superior for detection of GBS from vaginal-rectal specimens. Growth of non-GBS colonies required additional work to exclude the presence of GBS, especially after 48 h of incubation. Incubation of Select beyond 24 h did not significantly increase the yield of GBS. PMID:20962144

Rapid-viability PCR method for detection of live, virulent Bacillus anthracis in environmental samples.

PubMed

Létant, Sonia E; Murphy, Gloria A; Alfaro, Teneile M; Avila, Julie R; Kane, Staci R; Raber, Ellen; Bunt, Thomas M; Shah, Sanjiv R

2011-09-01

In the event of a biothreat agent release, hundreds of samples would need to be rapidly processed to characterize the extent of contamination and determine the efficacy of remediation activities. Current biological agent identification and viability determination methods are both labor- and time-intensive such that turnaround time for confirmed results is typically several days. In order to alleviate this issue, automated, high-throughput sample processing methods were developed in which real-time PCR analysis is conducted on samples before and after incubation. The method, referred to as rapid-viability (RV)-PCR, uses the change in cycle threshold after incubation to detect the presence of live organisms. In this article, we report a novel RV-PCR method for detection of live, virulent Bacillus anthracis, in which the incubation time was reduced from 14 h to 9 h, bringing the total turnaround time for results below 15 h. The method incorporates a magnetic bead-based DNA extraction and purification step prior to PCR analysis, as well as specific real-time PCR assays for the B. anthracis chromosome and pXO1 and pXO2 plasmids. A single laboratory verification of the optimized method applied to the detection of virulent B. anthracis in environmental samples was conducted and showed a detection level of 10 to 99 CFU/sample with both manual and automated RV-PCR methods in the presence of various challenges. Experiments exploring the relationship between the incubation time and the limit of detection suggest that the method could be further shortened by an additional 2 to 3 h for relatively clean samples.

A Demonstration of Nitrogen Dynamics in Oxic and Hypoxic Soils and Sediments.

ERIC Educational Resources Information Center

Ambler, Julie; Pelovitz, Kelly; Ladd, Timothy; Steucek, Guy

2001-01-01

Describes an experiment in which the incubation time to observe denitrification and other processes of the nitrogen cycle is reduced from 7-14 days to 24-48 hours. Presents calculations of processes in the nitrogen cycle in the form of a dichotomous key. (SAH)

Investigation of biosolids degradation under flooded environments for use in underwater cover designs for mine tailing remediation.

PubMed

Jia, Yu; Nason, Peter; Maurice, Christian; Alakangas, Lena; Öhlander, Björn

2015-07-01

To evaluate the potential suitability of digested sewage sludge (frequently termed biosolids) for use as underwater cover material for mine waste tailings, the degradability of biosolids at 20 - 22 °C under flooded anaerobic conditions was evaluated during incubation for 230 days. Leaching of elements from the flooded anaerobic system was also evaluated. Biosolid degradation was confirmed by the generation and accumulation of CH4 and CO2. Specifically, approximately 1.65 mmoL gas/g biosolids was generated as a result of incubation, corresponding to degradation of 7.68% of the organic matter, and the residue was stable at the end of the laboratory experiment. Under field conditions in northern Sweden, it is expected that the degradation rate will be much slower than that observed in the present study (Nason et al. Environ Earth Sci 70:30933105, 2013). Although the majority of biosolid fractions (>92%) were shown to be recalcitrant during the incubation period, long-term monitoring of further degradability of residue is necessary. The leaching results showed that most of the metals and metalloids leached from the biosolids at day 230 were below the limit value for non-hazardous waste, although Ni was the only element approximately three times higher than the limit value for inert material at the landfill site. In conclusion, biosolids have potential for use as covering material for underwater storage of tailings based on their biodegradability and leaching of elements.

Modification of nanocellulose by poly-lysine can inhibit the effect of fumonisin B1 on mouse liver cells.

PubMed

Jebali, Ali; Yasini Ardakani, Seyed Ali; Shahdadi, Hossein; Balal Zadeh, Mohammad Hossein; Hekmatimoghaddam, Seyedhossein

2015-02-01

Fumonisin B1 is an important mycotoxin, mainly produced by Fusarium verticillioides. It has toxic effects on liver, brain, and kidney cells. The first aim of this study was to synthesize nanocellulose modified with poly-lysine (NMPL), and the second aim was to evaluate the adsorption of fumonisin B1 by NMPL. As third aim, the function of mouse liver cells was investigated after exposure to fumonisin B1, and fumonisin B1+ NMPL. In this study, NMPL was prepared using cross-linker, and then incubated with fumonisin B1 at controlled conditions. After incubation, the adsorption and release of fumonisin B1 were evaluated in each condition. Next, mouse liver cells were separately exposed to fumonisin B1, NMPL, and (fumonisin B1+NMPL). Then, the level of aniline aminotransferase (ALT) and aspartate aminotransferase (AST) was evaluated. It was found that both adsorption and release of fumonisin B1 were not affected by temperature and incubation time, but affected by pH and concentration of NMPL. Also, this study showed NMPL could adsorb fumonisin B1 in different foodstuffs. Importantly, although the levels of ALT and AST were increased when the cells were treated with fumonisin B1 alone, they were not affected when exposed to NMPL or (fumonisin B1+NMPL). The authors suggest that NMPL is a good adsorbent to remove and inhibit fumonisin B1. Copyright © 2015 Elsevier B.V. All rights reserved.

Prior reproductive experience alters prolactin-induced macrophage responses in pregnant rats.

PubMed

Carvalho-Freitas, Maria Isabel Roth; Anselmo-Franci, Janete A; Palermo-Neto, João; Felicio, Luciano F

2013-09-01

Reproductive experience (i.e., pregnancy and lactation) induces physiological changes in mammals. A previous reproductive experience was recently shown to modulate the activity of dopaminergic hypothalamic systems while decreasing serum prolactin levels and oxidative burst activity in peritoneal macrophages. Dopamine receptor antagonists increase serum prolactin levels, and both prolactin and dopamine receptors may be involved in the modulation of macrophage activity, providing a means of communication between the nervous and immune systems. The present study evaluated the in vitro effects of prolactin and a dopamine D2 receptor antagonist on the peritoneal activity of macrophages from primigravid and multigravid female rats during the third trimester of pregnancy. Oxidative bursts and phagocytosis in peritoneal macrophages were evaluated by flow cytometry. Primigravid and multigravid Wistar rats, during the third trimester of pregnancy (i.e., days 17-21), were used. Peritoneal fluid samples from these rats were first incubated with prolactin (10 and 100 nM) for different periods of time. The same procedure was repeated to evaluate the effects of domperidone (10 and 100 nM) on macrophage activity. Our results showed that macrophages from multigravid rats responded more effectively to in vitro incubation with prolactin, especially with regard to the intensity and percentage of phagocytosis. Additionally, these effects were more pronounced after incubation periods of 30 min or 4 h. These data suggest that macrophages during a second pregnancy become more sensitive to the phagocytotic effects of prolactin. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Can serums be replaced by Mueller-Hinton agar in germ tube test?

PubMed

Atalay, M A; Koc, A N; Parkan, O M; Aydemir, G; Elmali, F; Sav, H

2017-01-01

The germ tube test (GTT) is inexpensive, easy, and well-defined test that differentiates Candida albicans (excluding Candida dubliniensis and Candida africana) from other species. The aim of this study was to evaluate various serums (i.e., human, rabbit, horse, and fetal bovine serum) used in the GTT and Mueller-Hinton agar (MHA). Fifty species isolated from various clinical samples that were defined as C. albicans by both conventional and DNA sequence analysis methods were included in the study. One to two colonies of C. albicans were mixed into 0.5-1 ml of fetal bovine serum, horse serum, rabbit serum, and human serum. Serums and MHA were incubated at 37°C for GTT. They were removed from the incubator and evaluated after 30 min, 1 h, 2 h, and 3 h of incubation. The GTT was accepted to be positive only if germ tube was 1/2 the width and 3 times the length of the parent yeast cell and with no constriction at the point of origin. When the use of serums and MHA for GTT was statistically evaluated, according to the positive scoring, the best results were obtained with MHA and with rabbit, horse, and fetal bovine serum, respectively. The best definition over time statistically was the third hour. It is suggested that inexpensive MHA is a fast, appropriate, and reliable medium for the probable diagnosis of GTT and C. albicans; however, additional studies are still needed to define other Candida species.

[Influence of physiologic 17 beta-estradiol concentrations on gene E6 expression in HVP type 18 in vitro].

PubMed

Dziubińska-Parol, Izabella; Gasowska, Urszula; Rzymowska, Jolanta; Kwaśniewska, Anna

2003-09-01

Many recent studies indicate that long term use of contraceptives is a strong risk factor in the development of cervical cancer. Steroid hormones, in persistent papilloma virus infection act on various levels, one of them is enhancing transforming activity of the virus. The aim of the study was to estimate if physiological concentrations of 17 beta-estradiol could influence expression of viral transforming genes. HeLa cell lines were incubated with three different physiological concentrations and and on the third day of incubation the level of E6 gene expression was determined. Results show no differences in expression between the control culter, and cultures incubated with physiological concentrations. It indicates that normal levels of 17 beta-estradiol don't play role in transforming process but it also shows need to analyse higher levels of hormones by quantitative analyses in prospective studies.

Nutrient fluxes across sediment-water interface in Bohai Bay Coastal Zone, China.

PubMed

Mu, Di; Yuan, Dekui; Feng, Huan; Xing, Fangwei; Teo, Fang Yenn; Li, Shuangzhao

2017-01-30

Sediment cores and overlying water samples were collected at four sites in Tianjin Coastal Zone, Bohai Bay, to investigate nutrient (N, P and Si) exchanges across the sediment-water interface. The exchange fluxes of each nutrient species were estimated based on the porewater profiles and laboratory incubation experiments. The results showed significant differences between the two methods, which implied that molecular diffusion alone was not the dominant process controlling nutrient exchanges at these sites. The impacts of redox conditions and bioturbation on the nutrient fluxes were confirmed by the laboratory incubation experiments. The results from this study showed that the nutrient fluxes measured directly from the incubation experiment were more reliable than that predicted from the porewater profiles. The possible impacts causing variations in the nutrient fluxes include sewage discharge and land reclamation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Influence of porcine spermadhesins on the susceptibility of boar spermatozoa to high dilution.

PubMed

Centurion, Fernando; Vazquez, Juan M; Calvete, Juan J; Roca, Jordi; Sanz, Libia; Parrilla, Inmaculada; Garcia, Eva M; Martinez, Emilio A

2003-08-01

The effect of heparin-binding and non-heparin-binding spermadhesins on the viability, motility, and mitochondrial activity of boar spermatozoa at the high dilution (300,000 sperm/ml) to which sperm are exposed during the process of sex sorting by flow cytometry was investigated. Incubation of spermatozoa with heparin-binding spermadhesins caused a time- and dose-dependent decrease in the percentage of functional spermatozoa. The percentage of viable spermatozoa incubated at 38 degrees C with heparin-binding spermadhesins diluted in PBS (1 mg/ml) dropped from 75% (0.5 h) to 4% (5 h), whereas the percentage of viable spermatozoa incubated in PBS without proteins (control) decreased from 85% (0.5 h) to 19% (5 h). Addition of non-heparin-binding PSP-I/PSP-II spermadhesin to the PBS resulted in a concentration-dependent increment of the percentage of viable cells (65% after 5-h incubation), with maximum effect at 1.5 mg/ml. The heparin-binding spermadhesins totally suppressed sperm motility and mitochondrial activity after 5 h of incubation. The same parameters of sperm incubated in the presence of 1.5 mg/ml of PSP-I/PSP-II were 50% and 58%, respectively, and the percentages of control sperm displaying motility and mitochondrial activity were 21% and 26%, respectively. Moreover, the viability, motility, and mitochondrial activity all decreased on incubation of spermatozoa with mixtures of PSP-I/PSP-II and heparin-binding spermadhesins as the concentration of the latter increased. We conclude that PSP-I/PSP-II and the heparin-binding spermadhesins exert antagonistic effects on the functionality of highly diluted boar spermatozoa. The finding that PSP-I/PSP-II contributes to maintaining sperm with high viability, motility, and mitochondrial activity for at least 5 h at physiological temperature points to its potential use as an additive for sperm preservation, specifically of highly diluted, flow-sorted spermatozoa for sex preselection.

Identification of apoptotic bodies in equine semen.

PubMed

Caselles, A B; Miro-Moran, A; Morillo Rodriguez, A; Gallardo Bolaños, J M; Ortega-Ferrusola, C; Salido, G M; Peña, F J; Tapia, J A; Aparicio, I M

2014-04-01

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37 °C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin-V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer-assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process. © 2014 Blackwell Verlag GmbH.

Evaluation of new antibiotic cocktails against contaminating bacteria found in allograft tissues.

PubMed

Serafini, Agnese; Riello, Erika; Trojan, Diletta; Cogliati, Elisa; Palù, Giorgio; Manganelli, Riccardo; Paolin, Adolfo

2016-12-01

Contamination of retrieved tissues is a major problem for allograft safety. Consequently, tissue banks have implemented decontamination protocols to eliminate microorganisms from tissues. Despite the widespread adoption of these protocols, few comprehensive studies validating such methods have been published. In this manuscript we compare the bactericidal activity of different antibiotic cocktails at different temperatures against a panel of bacterial species frequently isolated in allograft tissues collected at the Treviso Tissue Bank Foundation, a reference organization of the Veneto Region in Italy that was instituted to select, recover, process, store and distribute human tissues. We were able to identify at least two different formulations capable of killing most of the bacteria during prolonged incubation at 4 °C.

Mulberry anthocyanin biotransformation by intestinal probiotics.

PubMed

Cheng, Jing-Rong; Liu, Xue-Ming; Chen, Zhi-Yi; Zhang, You-Sheng; Zhang, Ye-Hui

2016-12-15

This study was designed to evaluate mulberry anthocyanins bioconversion traits for intestinal probiotics. Five intestinal beneficial bacteria were incubated with mulberry anthocyanins under anaerobic conditions at 37°C, and bacterial β-glucosidase activity and anthocyanin level were determined. Results demonstrated that all strains could convert mulberry anthocyanins to some extent. With high β-glucosidase production capacity, Streptococcus thermophiles GIM 1.321 and Lactobacillus plantarum GIM 1.35 degraded mulberry anthocyanins by 46.17% and 43.62%, respectively. Mulberry anthocyanins were mainly biotransformed to chlorogenic acid, crypto-chlorogenic acid, caffeic acid, and ferulic acid during the anaerobic process. Non-enzymatic deglycosylation of anthocyanins also occurred and approximately 19.42% of the anthocyanins were degraded within 48h by this method. Copyright © 2016 Elsevier Ltd. All rights reserved.

Sonic extracts from a bacterium related to periapical disease activate gelatinase A and inactivate tissue inhibitor of metalloproteinases TIMP-1 and TIMP-2.

PubMed

Sato, Y; Kishi, J; Suzuki, K; Nakamura, H; Hayakawa, T

2009-12-01

To examine the effects of sonicated bacterial extracts (SBEs) from three related to periapical disease bacteria (Porphyromonas gingivalis, P. endodontalis and F. nucleatum) on the activation of matrix metalloproteinase (MMP-2) and the inactivation of tissue inhibitors of metalloproteinase (TIMP-1 and TIMP-2). Each SBE was added to cultures of human periodontal ligament (PL) cells or HT1080 cells and their supernatants were analysed by zymography for MMP-2. Each SBE was added to PL cell cultures, and the amount of TIMP-1 was determined by ELISA. P. gingivalis SBE was incubated with HT1080 cell culture supernatants, and the amounts of TIMP-1 and TIMP-2 were determined by ELISA. Statistical analysis was performed with the paired Student's t-test. In extracts of PL cells that had been incubated in the presence of P. gingivalis SBE, one representing pro-MMP-2 (72 kDa) and a band corresponding to the active MMP-2 (66 kDa) were observed; but in the other extracts it was not detected. When HT1080 cells were treated with P. gingivalis SBE, the pro-MMPs was processed into 86- and 66-kDa fragments, but in the other extracts, the processing did not occur when the other SBEs were used. When PL cells were incubated with the same SBEs, the amount of TIMP-1 was markedly decreased (P < 0.01), but in the other extracts, it was not. The amounts of both TIMP-1 and TIMP-2 were decreased in a dose-dependent manner when HT1080 cell culture supernatant was incubated with P. gingivalis SBE. These findings suggest that P. gingivalis SBE may cause connective tissue to be destroyed, contributing to the process of periapical disease, by activating pro-MMP-2 as well as by inactivating TIMP-1 and TIMP-2.

An Effect of Dexamethasone on Adenosine 3′,5′ -Monophosphate Content and Adenosine 3′,5′ -Monophosphate Phosphodiesterase Activity of Cultured Hepatoma Cells

PubMed Central

Manganiello, Vincent; Vaughan, Martha

1972-01-01

The effect of dexamethasone on adenosine 3′,5′-monophosphate (cAMP) phosphodiesterase activity in cultured HTC hepatoma cells was investigated. Homogenates of these cells contain phosphodiesterase activity with two apparent Michaelis constants for cAMP (2-5 μm and 50 μm). At all substrate concentrations tested, phosphodiesterase activity was decreased 25-40% in cells incubated for 36 hr or more with 1 μm dexamethasone. Acid phosphatase activity in the same cells was not decreased. α-Methyl testosterone, 1 μm, was without effect on phosphodiesterase activity. Incubation for 10 min with epinephrine plus theophylline increased the cAMP content of the HTC cells 3- to 6-fold. In cells incubated for 72 hr with dexamethasone, the basal concentration of cAMP was slightly increased and the increment produced by epinephrine plus theophylline was markedly increased. We believe that in many cells the so-called permissive effects of steroid hormones on cAMP mediated processes may be due to an effect of these hormones on cAMP phosphodiesterase activity similar to that observed in HTC cells incubated with dexamethasone. PMID:4341439

Measuring temperature dependence of soil respiration: importance of incubation time, soil type, moisture content and model fits

NASA Astrophysics Data System (ADS)

Schipper, L. A.; Robinson, J.; O'Neill, T.; Ryburn, J.; Arcus, V. L.

2015-12-01

Developing robust models of the temperature response and sensitivity of soil respiration is critical for determining changes carbon cycling in response to climate change and at daily to annual time scales. Currently, approaches for measuring temperature dependence of soil respiration generally use long incubation times (days to weeks and months) at a limited number of incubation temperatures. Long incubation times likely allow thermal adaptation by the microbial population so that results are poorly representative of in situ soil responses. Additionally, too few incubation temperatures allows for the fit and justification of many different predictive equations, which can lead to inaccuracies when used for carbon budgeting purposes. We have developed a method to rapidly determine the response of soil respiration rate to wide range of temperatures. An aluminium block with 44 sample slots is heated at one end and cooled at the other to give a temperature gradient from 0 to 55°C at about one degree increments. Soil respiration is measured within 5 hours to minimise the possibility of thermal adaptation. We have used this method to demonstrate the similarity of temperature sensitivity of respiration for different soils from the same location across seasons. We are currently testing whether long-term (weeks to months) incubation alter temperature response and sensitivity that occurs in situ responses. This method is also well suited for determining the most appropriate models of temperature dependence and sensitivity of soil respiration (including macromolecular rate theory MMRT). With additional testing, this method is expected to be a more reliable method of measuring soil respiration rate for soil quality and modelling of soil carbon processes.

Osteoblast-like cell response to macro- and micro-patterned carbon scaffolds obtained from the sea rush Juncus maritimus.

PubMed

López-Álvarez, M; Pereiro, I; Serra, J; de Carlos, A; González, P

2011-08-01

Carbon scaffolds with a directional patterned surface were obtained by pyrolysis of the sea rush Juncus maritimus. The structure of the scaffolds was investigated using scanning electron microscopy, mercury porosimetry and interferometric profilometry. X-ray diffraction and x-ray fluorescence were the techniques used for their chemical characterization. The alignment and differentiation of pre-osteoblasts (MC3T3-E1 cell line) incubated on the patterned scaffolds were evaluated by scanning electron microscopy, confocal laser scanning microscopy and by the quantification of the phosphatase alkaline activity and the osteocalcin synthesis. It was found that pyrolysis at 500 °C preserved and even enhanced the natural macro- and micro-patterning of the plant. The results obtained for porosity and chemical composition validated these structures as viable scaffolds for tissue engineering applications. Finally, the patterned surface was confirmed to promote the oriented growth of the pre-osteoblasts MC3T3-E1, not only after short periods of incubation (hours) but also after longer ones (several weeks). The quantification of the cell differentiation markers together with the evaluation of the cell layer morphology up to 28 days of incubation confirmed the differentiation of MC3T3-E1 cells to osteoblasts. © 2011 IOP Publishing Ltd

Evaluating acetaldehyde synthesis from L-/sup 14/C(U)) threonine by Streptococcus thermophilus and Lactobacillus bulgaricus

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wilkins, D.W.; Schmidt, R.H.; Shireman, R.B.

To evaluate the synthesis of acetaldehyde from threonine during growth of yogurt cultures, Streptococcus thermophilus MS1 and Lactobacillus bulgaricus MR1 were grown in defined medium in which 10% of the total threonine was composed of L-(carbon-14(U))threonine. Acetaldehyde production was monitored by formation of 2,4-dinitrophenylhydrazone followed by separation and analysis using high performance liquid chromatography. After growth for 8 h at 42/sup 0/C, approximately 2.0% of the total acetaldehyde (780.4 nmol) produced was from L-(carbon-14)threonine. Threonine aldolase activity was determined in cell-free extracts from S. thermophilus and L. bulgaricus grown in Elliker broth. Increasing incubation temperature from 30 to 42/sup 0/Cmore » decreased threonine aldolase activity in cells of the streptococcus harvested after 8 h of incubation. Effect of incubation temperature was more dramatic in cells harvested after 18 h where the activity of cells grown at 48/sup 0/C was 89% lower than that of cells grown at 30/sup 0/C. Cell extracts from S. thermophilus MS1 possessed higher threonine aldolase activity than did those from L. bulgaricus MR1. Increased assay temperature from 30 to 42/sup 0/C increased threonine aldolase activity in S. thermophilus MS1.« less

Comparative study of the in vitro activity of various antifungal drugs against Scedosporium spp. in aerobic and hyperbaric atmosphere versus normal atmosphere.

PubMed

Farina, C; Marchesi, G; Passera, M; Diliberto, C; Russello, G

2012-06-01

Scedosporium spp. have been observed with increasing frequency over the last decade in immunocompromised patients and trauma patients. This mould is often multi-drug resistant and its mortality rate remains very high. The primary goal of this study was to obtain data concerning the in vitro susceptibility of 13 Scedosporium strains comparing the in vitro incubation in aerobic versus hyperbaric conditions. Chemosensitivity of thirteen Scedosporium strains was evaluated after a 72h-incubation in a normoxic (21% O2) normobaric (1 ATA) atmosphere versus a hyperoxic (100% O2) hyperbaric (2-3 ATA), and after a re-incubation at room temperature for an additional 72h. All S. apiospermum and S. prolificans strains showed no growth after incubation in hyperbaric hyperoxic atmosphere. However, when plates were then maintained at room temperature in aerobic conditions, growth was systematically observed from 36 to 96h, and Minimal inhibitory concentration (MIC) values were the same obtained after incubation in aerobic conditions. These results suggest impressive in vitro fungistatic activity of the hyperoxic hyperbaric atmosphere, even if its effect is strictly time-dependent. This preliminary in vitro study has potential clinical relevance because it focuses on examining in vitro combination therapy using hyperoxic hyperbaric conditions plus a single antifungal agent, rather than using combinations of different antifungal drugs, to potentially increase the antifungal activity. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

Factors affecting the recovery of Legionella pneumophila serogroup 1 from cooling tower water systems.

PubMed

Lu, H F; Tsou, M F; Huang, S Y; Tsai, W C; Chung, J G; Cheng, K S

2001-09-01

A total of 20 water samples collected from the cooling towers at 20 different sites were analyzed under various conditions for the presence of Legionella pneumophila serogroup 1. A comparative assessment was performed to evaluate methods of sample collection (spray drops, beneath water at 20- to 40-cm depth, and water outlet), concentration (filtration and centrifugation), acid buffer treatment (no treatment, treatment for 3, 5, and 15 min), and CO2 incubation or candle jar incubation. The reduction in viable colonies and false negative rate were compared for the different factors. No quantitative differences in isolation of L. pneumophila serogroup 1 was found among samples collected from water at a depth of 20 to 40 cm, from water outlet, and from spray drops. Treatment in an acid buffer for 15 min significantly reduced the recovery rate, with a reduction in bacterial counts of about 40%, compared with a 3-min (12%) or a 5-min (25%) treatment. Acid buffer treatment for 3 or 5 min reduced the overgrowth of commensal flora. This treatment improved the selectivity but not the sensitivity for L. pneumophila serogroup 1. Colonies on plates incubated at 37 degrees C in a candle jar with a humidified atmosphere grew better than those incubated at 35 degrees C with 5% CO2. These results demonstrate that methods of sample collection, concentration, and incubation, but not collection site, can affect the isolation rate for L. pneumophila serogroup 1.

Weight at weaning of preterm infants from incubator to bassinet: a randomized clinical trial.

PubMed

Berger, Irit; Marom, Ronella; Mimouni, Francis; Kopelovich, Rosalia; Dollberg, Shaul

2014-06-01

To evaluate thermal responses, weight gain, and resting energy expenditure (REE) in preterm infants weaned from an infant incubator to a warming bassinet after reaching a body weight of either 1,500 or 1,600 g, respectively. Preterm infants weighing ≤ 1,500 g were randomly allocated for weaning from a convective incubator after reaching 1,500 g body weight (study group) or 1,600 g (controls). Postweaning body temperature was recorded at short intervals. REE was measured before and after weaning, while the weight gain was calculated daily. All 21 enrolled infants were weaned successfully to a bassinet (room temperature). REE inside the incubator and in the warming bassinette was similar between infants weaned at 1,500 g and at 1,600 g (2.9 ± 6.8 vs. 1.1 ± 4.8 kcal/kg/d, respectively (p = 0.7). REE increased slightly after weaning in both groups. Weight gain and days from randomization to discharge did not differ between groups. Weaning very low birth weight infants from an incubator to a warming bassinet at a body weight of 1,500 g is feasible and has no significant deleterious effects on weight gain and REE. The many benefits provided by such greater accessibility to premature infants are discussed. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Computer assisted sperm analysis of motility patterns of postthawed epididymal spermatozoa of springbok (Antidorcas marsupialis), impala (Aepyceros melampus), and blesbok (Damaliscus dorcus phillipsi) incubated under conditions supporting domestic cattle in vitro fertilization.

PubMed

Chatiza, F P; Bartels, P; Nedambale, T L; Wagenaar, G M

2012-07-15

The need for information on the reproductive physiology of different wildlife species is important for ex situ conservation using such methods as in vitro fertilization (IVF). Information on species reproductive physiology and evaluation of sperm quality using accurate, objective, repeatable methods, such as computer-assisted sperm analysis (CASA) for ex situ conservation has become a priority. The aim of this study was to evaluate motility patterns of antelope epididymal spermatozoa incubated for 4 h under conditions that support bovine IVF using CASA. Cauda epididymal spermatozoa were collected postmortem from testicles of springbok (N=38), impala (N=26), and blesbok (N=42), and cryopreserved in biladyl containing 7% glycerol. Spermatozoa were thawed and incubated in Capacitation media and modified Tyrode lactate (m-TL) IVF media using a protocol developed for domestic cattle IVF. The study evaluates 14 motility characteristics of the antelope epididymal sperm at six time points using CASA. Species differences in CASA parameters evaluated under similar conditions were observed. Several differences in individual motility parameters at the time points were reported for each species. Epididymal sperm of the different antelope species responded differently to capacitation agents exhibiting variations in hyperactivity. Motility parameters that describe the vigor of sperm decreased over time. Spermatozoa from the different antelope species have different physiological and optimal capacitation and in vitro culture requirements. The interspecies comparison of kinematic parameters of spermatozoa between the antelopes over several end points contributes to comparative sperm physiology which forms an important step in the development of species specific assisted reproductive techniques (ARTs) for ex situ conservation of these species. Copyright © 2012 Elsevier Inc. All rights reserved.

Quantitative evaluation of ruminal methane and carbon dioxide formation from formate through C-13 stable isotope analysis in a batch culture system.

PubMed

He, Z X; Qiao, J Y; Yan, Q X; Tan, Z L; Wang, M

2018-04-12

Methane produced from formate is one of the important methanogensis pathways in the rumen. However, quantitative information of CH4 production from formate has been rarely reported. The aim of this study was to characterize the conversion rate (CR) of formic acid into CH4 and CO2 by rumen microorganisms. Ground lucerne hay was incubated with buffered ruminal fluid for 6, 12, 24 and 48 h. Before the incubation, 13C-labeled H13COOH was also supplied into the incubation bottle at a dose of 0, 1.5, 2.2 or 2.9 mg/g of DM substrate. There were no interactions (P>0.05) between dose and incubation time for all variables evaluated. When expressed as an absolute amount (ml in gas sample) or a relative CR (%), both 13CH4 and 13CO2 production quadratically increased (P<0.01) with the addition of H13COOH. The total 13C (13CH4 and 13CO2) CR was also quadratically increased (P<0.01) when H13COOH was added. Moreover, formate addition linearly decreased (P<0.031) the concentrations of NH3-N, total and individual volatile fatty acids (acetate, propionate and butyrate), and quadratically decreased (P<0.014) the populations of protozoa, total methanogens, Methanosphaera stadtmanae, Methanobrevibacter ruminantium M1, Methanobrevibacter smithii and Methanosarcina barkeri. In summary, formate affects ruminal fermentation and methanogenesis, as well as the rumen microbiome, in particular microorganisms which are directly or indirectly involved in ruminal methanogenesis. This study provides quantitative verification for the rapid dissimilation of formate into CH4 and CO2 by rumen microorganisms.

Cryohemolysis, erythrocyte osmotic fragility, and supplementary hematimetric indices in the diagnosis of hereditary spherocytosis.

PubMed

Emilse, Ledesma Achem Miryam; Cecilia, Haro; María, Terán Magdalena; Eugenia, Mónaco María; Alicia, Issé Blanca; Lazarte, Sandra Stella

2018-03-01

Hereditary spherocytosis (HS) is a chronic hemolytic anemia characterized by microspherocytes in the peripheral blood and increased erythrocyte osmotic fragility (EOF). This study evaluated the cryohemolysis test (CHT); initial hemolysis (IH); immediate and incubated hemolysis percentage in 5.5 g/L NaCl (H5.5); mean corpuscular hemoglobin concentration (MCHC); red blood cell distribution width (RDW); and Hb/MCHC, Hb/RDW, and MCHC/RDW ratios for the diagnosis of HS. Data from 13 patients with HS were evaluated at the Instituto de Bioquímica Aplicada and compared with data from 14 unaffected individuals and 11 patients with anemia due to another etiology. Total blood and reticulocyte counts, CHT, and immediate and incubated EOF were performed in all subjects; sensitivity, specificity, efficiency, and Youden index (YI) were calculated. Eight patients with HS had MCHC ≥345 g/L, 10 had RDW ≥14.5%, 12 had IH >5.0 g/L, 11 had immediate H5.5 ≥5%, and 13 had incubated H5.5 ≥50% (the cut-off value to consider HS). The efficiency and YI were: immediate H5.5 (0.94-0.85), incubated H5.5 (0.89-0.82), IH (0.89-0.78), MCHC (0.87-0.62), CHT (0.84-0.54), and Hb/MCHC (0.71-0.56), respectively. The calculated ratios could distinguish subjects with HS from unaffected individuals ( P <0.05), but not those with anemia of another etiology ( P >0.05). Although the CHT and supplementary hematimetric indexes were useful to differentiate individuals with SH from healthy controls, they cannot distinguish from anemias of other etiology. CHT and MCHC, in addition to EOF, are recommended for diagnosing HS patients because of their low cost and efficiency.

D-Aspartic acid and nitric oxide as regulators of androgen production in boar testis.

PubMed

Lamanna, Claudia; Assisi, Loredana; Vittoria, Alfredo; Botte, Virgilio; Di Fiore, Maria Maddalena

2007-01-15

D-Aspartic acid (D-Asp) and nitric oxide (NO) are two biologically active molecules playing important functions as neurotransmitters and neuromodulators of nerve impulse and as regulators of hormone production by endocrine organs. We studied the occurrence of D-Asp and NO as well as their effects on testosterone synthesis in the testis of boar. This model was chosen for our investigations because it contains more Leydig cells than other mammals. Indirect immunofluorescence applied to cryostat sections was used to evaluate the co-localization of D-Asp and of the enzyme nitric oxide synthase (NOS) in the same Leydig cells. D-Asp and NOS often co-existed in the same Leydig cells and were found, separately, in many other testicular cytotypes. D-Asp level was dosed by an enzymatic method performed on boar testis extracts and was 40+/-3.6 nmol/g of fresh tissue. NO measurement was carried out using a biochemical method by NOS activity determination and expressed as quantity of nitrites produced: it was 155.25+/-21.9 nmol/mg of tissue. The effects of the two molecules on steroid hormone production were evaluated by incubating testis homogenates, respectively with or without D-Asp and/or the NO-donor L-arginine (L-Arg). After incubation, the testosterone presence was measured by immunoenzymatic assay (EIA). These in vitro experiments showed that the addition of D-Asp to incubated testicular homogenates significantly increased testosterone concentration, whereas the addition of L-Arg decreased the hormone production. Moreover, the inclusion of L-Arg to an incubation medium of testicular homogenates with added D-Asp, completely inhibited the stimulating effects of this enantiomer. Our results suggest an autocrine action of both D-Asp and NO on the steroidogenetic activity of the Leydig cell.

Broiler embryo bone development is influenced by incubator temperature, oxygen concentration and eggshell conductance at the plateau stage in oxygen consumption.

PubMed

Oviedo-Rondón, E O; Small, J; Wineland, M J; Christensen, V L; Mozdziak, P S; Koci, M D; Funderburk, S V L; Ort, D T; Mann, K M

2008-11-01

1. Four experiments were conducted to evaluate the effects of temperature (TEM) and oxygen (O(2)) concentrations during the last 4 d of incubation on bone development. Fertile eggs from two strains were obtained that either exhibited Low or High eggshell conductance (G). 2. Four experimental cabinets provided either four TEM (36, 37, 38 or 39 degrees C) or four O(2) concentrations (17, 19, 21 or 23% O(2)). Data were analysed as a 2 x 2 factorial design. In the fourth experiment, two temperatures (36 and 39 degrees C), two O(2) concentrations (17 and 23%) and the same Low and High G strains were evaluated in a 2 x 2 x 2 factorial design. 3. Body weights (BW) and residual yolks were obtained, both legs were dissected. Femur, tibia and shank weights, length and thickness were recorded. Relative asymmetry (RA) of each leg section was calculated. 4. The results indicated that elevated TEM during incubation increased RA between the two legs, mainly in the Low G strain. Chickens at the lowest O(2) concentrations had lighter and shorter tibias, lighter shanks, and increased RA of femur length compared to chickens in the 23% O(2). In the fourth experiment no interactions were observed between O(2) and TEM. High TEM depressed BW of Low G broilers, but no significant effect of treatments was observed on BW of High G broilers. Nevertheless, the high TEM or low O(2) independently caused reduced femur and tibia weights and length, shank length and thickness, and both low O(2) and high TEM together increased RA in shank weight. 5. These results suggest that late incubation conditions affect long bone development in broilers.

Bilirubin glucuronidation revisited: proper assay conditions to estimate enzyme kinetics with recombinant UGT1A1.

PubMed

Zhou, Jin; Tracy, Timothy S; Remmel, Rory P

2010-11-01

Bilirubin, an end product of heme catabolism, is primarily eliminated via glucuronic acid conjugation by UGT1A1. Impaired bilirubin conjugation, caused by inhibition of UGT1A1, can result in clinical consequences, including jaundice and kernicterus. Thus, evaluation of the ability of new drug candidates to inhibit UGT1A1-catalyzed bilirubin glucuronidation in vitro has become common practice. However, the instability of bilirubin and its glucuronides presents substantial technical challenges to conduct in vitro bilirubin glucuronidation assays. Furthermore, because bilirubin can be diglucuronidated through a sequential reaction, establishment of initial rate conditions can be problematic. To address these issues, a robust high-performance liquid chromatography assay to measure both bilirubin mono- and diglucuronide conjugates was developed, and the incubation conditions for bilirubin glucuronidation by human embryonic kidney 293-expressed UGT1A1 were carefully characterized. Our results indicated that bilirubin glucuronidation should be assessed at very low protein concentrations (0.05 mg/ml protein) and over a short incubation time (5 min) to assure initial rate conditions. Under these conditions, bilirubin total glucuronide formation exhibited a hyperbolic (Michaelis-Menten) kinetic profile with a K(m) of ∼0.2 μM. In addition, under these initial rate conditions, the relative proportions between the total monoglucuronide and the diglucuronide product were constant across the range of bilirubin concentration evaluated (0.05-2 μM), with the monoglucuronide being the predominant species (∼70%). In conclusion, establishment of appropriate incubation conditions (i.e., very low protein concentrations and short incubation times) is necessary to properly characterize the kinetics of bilirubin glucuronidation in a recombinant UGT1A1 system.

Effects of hydrostatic pressure on microbial alteration of sinking fecal pellets

NASA Astrophysics Data System (ADS)

Tamburini, Christian; Goutx, Madeleine; Guigue, Catherine; Garel, Marc; Lefèvre, Dominique; Charrière, Bruno; Sempéré, Richard; Pepa, Stéphane; Peterson, Michael L.; Wakeham, Stuart; Lee, Cindy

2009-08-01

We used a new experimental device called PASS (PArticle Sinking Simulator) during MedFlux to simulate changes in in situ hydrostatic pressure that particles experience sinking from mesopelagic to bathypelagic depths. Particles, largely fecal pellets, were collected at 200 m using a settling velocity NetTrap (SV NetTrap) in Ligurian Sea in April 2006 and incubated in high-pressure bottles (HPBs) of the PASS system under both atmospheric and continuously increasing pressure conditions, simulating the pressure change experienced at a sinking rate of 200 m d -1. Chemical changes over time were evaluated by measuring particulate organic carbon (POC), carbohydrates, transparent exopolymer particles (TEP), amino acids, lipids, and chloropigments, as well as dissolved organic carbon (DOC) and dissolved carbohydrates. Microbial changes were evaluated microscopically, using diamidinophenylindole (DAPI) stain for total cell counts and catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH) for phylogenetic distinctions. Concentrations (normalized to POC) of particulate chloropigments, carbohydrates and TEP decreased under both sets of incubation conditions, although less under the increasing pressure regime than under atmospheric conditions. By contrast, dissolved carbohydrates (normalized to DOC) were higher after incubation and significantly higher under atmospheric conditions, suggesting they were produced at the expense of the particulate fraction. POC-normalized particulate wax/steryl esters increased only under pressure, suggesting biochemical responses of prokaryotes to the increasing pressure regime. The prokaryotic community initially consisted of 43% Bacteria, 12% Crenarchaea and 11% Euryarchaea. After incubation, Bacteria dominated (˜90%) the prokaryote community in all cases, with γ- Proteobacteria comprising the greatest fraction, followed by the Cytophaga-Flavobacter cluster and α -Proteobacteria group. Using the PASS system, we obtained chemical and microbial evidence that degradation by prokaryotes associated with fecal pellets sinking through mesopelagic waters is limited by the increasing pressure they experience.

Association Between Fungal Contamination and Eye Bank-Prepared Endothelial Keratoplasty Tissue: Temperature-Dependent Risk Factors and Antifungal Supplementation of Optisol-Gentamicin and Streptomycin.

PubMed

Brothers, Kimberly M; Shanks, Robert M Q; Hurlbert, Susan; Kowalski, Regis P; Tu, Elmer Y

2017-11-01

Fungal contamination and infection from donor tissues processed for endothelial keratoplasty is a growing concern, prompting analysis of donor tissues after processing. To determine whether eyebank-processed endothelial keratoplasty tissue is at higher risk of contamination than unprocessed tissue and to model eyebank processing with regard to room temperature exposure on Candida growth in optisol-gentamicin and streptomycin (GS) with and without antifungal supplementation. An examination of the 2013 Eversight Eyebank Study follow-up database for risk factors associated with post-keratoplasty infection identified an increased risk of positive fungal rim culture results in tissue processed for endothelial keratoplasty vs unprocessed tissue. Processing steps at room temperature were hypothesized as a potential risk factor for promotion of fungal growth between these 2 processes. Candida albicans, Candida glabrata, and Candida parapsilosis endophthalmitis isolates were each inoculated into optisol-GS and subjected to 2 different room temperature incubation regimens reflective of current corneal tissue handling protocols. Eversight Eyebank Study outcomes and measures were follow-up inquiries from 6592 corneal transplants. Efficacy study outcomes and measures were fungal colony-forming units from inoculated vials of optisol-GS taken at 2 different processing temperatures. Donor rim culture results were 3 times more likely to be positive for fungi in endothelial keratoplasty-processed eyes (1.14%) than for other uses (0.37%) (difference, 0.77%; 95% CI, 0.17-.1.37) (P = .009). In vitro, increased room temperature incubation of optisol-GS increased growth of Candida species over time. The addition of caspofungin and voriconazole decreased growth of Candida in a species-dependent manner. Detectable Candida growth in donor rim cultures, associated with a higher rate of post keratoplasty infection, is seen in endothelial keratoplasty tissue vs other uses at the time of transplantation, likely owing in part to eyebank preparation processes extending the time of tissue warming. Reduced room temperature incubation and the addition of antifungal agents decreased growth of Candida species in optisol-GS and should be further explored to reduce the risk of infection.

Design of fiber optic based respiratory sensor for newborn incubator application

NASA Astrophysics Data System (ADS)

Dhia, Arika; Devara, Kresna; Abuzairi, Tomy; Poespawati, N. R.; Purnamaningsih, Retno W.

2018-02-01

This paper reports the design of respiratory sensor using fiber optic for newborn incubator application. The sensor works based on light intensity losses difference obtained due to thorax movement during respiration. The output of the sensor launched to support electronic circuits to be processed in Arduino Uno microcontroler such that the real-time respiratory rate (breath per minute) can be presented on LCD. Experiment results using thorax expansion of newborn simulator show that the system is able to measure respiratory rate from 10 up to 130 breaths per minute with 0.595% error and 0.2% hysteresis error.

Erosion of composite materials

NASA Technical Reports Server (NTRS)

Springer, G. S.

1980-01-01

A model for describing the response of uncoated and coated fiber reinforced composites subjected to repeated impingements of liquid (rain) droplets is presented. The model is based on the concept that fatigue is the dominant factor in the erosion process. Algebraic expressions are provided which give the incubation period, the rate of mass loss past the incubation period, and the total mass loss of the material during rain impact. The influence of material properties on erosion damage and the protection offered by different coatings are discussed and the use of the model in the design in the design of structures and components is illustrated.

The effects of brefeldin-A on the high mannose oligosaccharides of mouse thyrotropin, free alpha-subunits, and total glycoproteins.

PubMed

Perkel, V S; Liu, A Y; Miura, Y; Magner, J A

1988-07-01

We have studied the effects of Brefeldin-A (BFA) on the processing of high mannose (Man) oligosaccharides of TSH. BFA is a drug that inhibits the intracellular translocation of newly synthesized glycoproteins and causes dilatation of the rough endoplasmic reticulum (RER) as well as mild swelling of the Golgi apparatus. Mouse pituitary thyrotropic tumor tissue was incubated with [3H]Man for a 2-h pulse, with and without a 3-h chase; BFA (5 micrograms/ml) was included during selected pulse and selected chase incubations. TSH and free alpha-subunits were obtained from detergent lysates of tissue by immunoprecipitation using specific antisera. Total glycoproteins were obtained by trichloroacetic acid precipitation. Endoglycosidase-H-released [3H]oligosaccharides were analyzed by paper chromatography. BFA inhibited carbohydrate processing of TSH, free alpha-subunits, and total glycoproteins, resulting in the accumulation of Man8GlcNAc2, Man7GlcNAc2, Man6GlcNAc2, and Man5GlcNAc2, especially during the chase period. Subcellular fractions enriched in RER, heavy (proximal) Golgi, and light (distal) Golgi were prepared by centrifugation in discontinuous sucrose gradients. [3H]Man-labeled oligosaccharides of TSH and total glycoproteins in the subcellular fractions were analyzed. In contrast to oligosaccharides with eight or nine Man residues found in control incubations, BFA caused the accumulation of oligosaccharides containing five to eight Man residues. These BFA-induced oligosaccharide alterations began in the RER and proximal Golgi with the 2-h pulse and extended into the distal Golgi during the chase incubations. Thus, BFA blocks the normal intracellular transport and processing of TSH, free alpha-subunits, and total glycoproteins within thyrotrophs, causing species with smaller than normal high Man oligosaccharides to appear in subcellular compartments as early as the RER. The translocation block between RER and Golgi produced by BFA may prevent the processing of Man8GlcNAc2 to Man5GlcNAc2 by Golgi (alpha,1-2)mannosidase I, yet the species retained within the RER may be subject to ongoing processing by endoplasmic reticulum (alpha,1-2)mannosidase, resulting in the accumulation of Man5-8GlcNAc2 within the RER.

Regulation of breeding behavior: do energy-demanding periods induce a change in prolactin or corticosterone baseline levels in the common tern (Sterna hirundo)?

PubMed

Riechert, Juliane; Chastel, Olivier; Becker, Peter H

2014-01-01

Hormones are involved in reproductive decisions, linking environmental cues and body condition and adapting behavior. Mass loss is often accompanied by decreased prolactin and increased corticosterone concentrations, influencing incubation and brooding behavior and ultimately triggering nest desertion. Using blood-sucking bugs (Dipetalogaster maxima), we measured baseline prolactin, corticosterone, and ketone body values in incubating common terns (Sterna hirundo) between 2006 and 2009 during energy-demanding periods: 50 pairs were sampled hungry (after an incubation bout) and again fed (after foraging). In a second approach, we sampled 57 other pairs (experienced and inexperienced birds) three times over their individual breeding period, because reproduction, especially chick rearing, is a very energy-demanding process. In line with the common physiological pattern of fasting, we found significantly lower baseline prolactin values in hungry terns, which were negatively related to mass loss over the incubation bout, whereas corticosterone and ketone body levels were marginally increased. Compared to that in the incubation phase, the prolactin level dropped after hatching of chicks in inexperienced birds, perhaps indicating lower parental expenditure. Corticosterone, on the other hand, increased after hatching in males, probably linked to higher foraging activity, as males mainly deliver food during the first days. These energy-demanding periods clearly influenced hormones and ketone bodies, maybe reinforced by the low energy margin of this small seabird species, but energy reserves were not depleted to a level affecting behavior or reproductive success.

Regulation of amino acid transport across intestines of goldfish acclimatized to different environmental temperatures

PubMed Central

Mepham, T. B.; Smith, M. W.

1966-01-01

1. Serosal transfers of valine and threonine were measured using everted sacs of anterior intestine taken from goldfish acclimatized to different temperatures. 2. Both valine and threonine were actively transported at incubation temperatures equal to or greater than the previous environmental temperature of the fish. There was also a positive serosal transfer of valine, but not threonine, at incubation temperatures below the previous environmental temperature of the fish. 3. The mean stable transmural potentials and amino-acid-evoked potentials depended both on the temperature to which the fish had been acclimatized and on the temperature at which the sacs were incubated. 4. There was a linear relation between the transmural potential and the serosal transfer of amino acid, one additional μmole of valine or threonine being transferred/2 hr incubation period for each 3 mV rise in potential. There was a less obvious correlation between the amino-acid-evoked potential and on serosal transfer of amino acid. 5. Acclimatization of the goldfish intestine from 8 to 25° C, assessed by changes occurring in the transmural potential and serosal transfer of amino acids, tended to stabilize both parameters, but the compensation in each case was only partial. 6. It is possible that the imbalance in transfer of valine-like and threonine-like amino acids, seen at incubation temperatures below the previous acclimatization temperature of the fish, has a special function in initiating the process of acclimatization to the new environmental temperature. PMID:5972157

Hybrid Contactless Heating and Levitation

NASA Technical Reports Server (NTRS)

Lee, M. C.

1985-01-01

Acoustic and electromagnetic fields applied. In contactless processing apparatus, acoustic and electromagnetic levitating fields employed alternately or simultaneously with amplitude of each controlled to produce various combinations of heating, cooling, and levitation. Apparatus provides rapid heating and cooling or slow heating and cooling for such processes as nucleation, crystallization, incubation, deep undercooling, and heterogeneity control.

Inebriation and Inspiration? A Review of the Research on Alcohol and Creativity.

ERIC Educational Resources Information Center

Norlander, Torsten

1999-01-01

A review of research on the correlation between the intake of alcohol and creativity hypothesizes that modest alcohol consumption inhibits aspects of creativity based mainly on the secondary process (preparation, certain parts of illumination, and verification), and disinhibits those based mainly on the primary process (incubation, certain parts…

The Point of Creative Frustration and the Creative Process: A New Look at an Old Model.

ERIC Educational Resources Information Center

Sapp, D. David

1992-01-01

This paper offers an extension of Graham Wallas' model of the creative process. It identifies periods of problem solving, incubation, and growth with specific points of initial idea inception, creative frustration, and illumination. Responses to creative frustration are described including denial, rationalization, acceptance of stagnation, and new…

Accelerating bacterial growth detection and antimicrobial susceptibility assessment in integrated picoliter droplet platform.

PubMed

Kaushik, Aniruddha M; Hsieh, Kuangwen; Chen, Liben; Shin, Dong Jin; Liao, Joseph C; Wang, Tza-Huei

2017-11-15

There remains an urgent need for rapid diagnostic methods that can evaluate antibiotic resistance for pathogenic bacteria in order to deliver targeted antibiotic treatments. Toward this end, we present a rapid and integrated single-cell biosensing platform, termed dropFAST, for bacterial growth detection and antimicrobial susceptibility assessment. DropFAST utilizes a rapid resazurin-based fluorescent growth assay coupled with stochastic confinement of bacteria in 20 pL droplets to detect signal from growing bacteria after 1h incubation, equivalent to 2-3 bacterial replications. Full integration of droplet generation, incubation, and detection into a single, uninterrupted stream also renders this platform uniquely suitable for in-line bacterial phenotypic growth assessment. To illustrate the concept of rapid digital antimicrobial susceptibility assessment, we employ the dropFAST platform to evaluate the antibacterial effect of gentamicin on E. coli growth. Copyright © 2017 Elsevier B.V. All rights reserved.

Degradation of milk-based bioactive peptides by yogurt fermentation bacteria.

PubMed

Paul, M; Somkuti, G A

2009-09-01

To analyse the effect of cell-associated peptidases in yogurt starter culture strains Lactobacillus delbrueckii ssp. bulgaricus (LB) and Streptococcus thermophilus (ST) on milk-protein-based antimicrobial and hypotensive peptides in order to determine their survival in yogurt-type dairy foods. The 11mer antimicrobial and 12mer hypotensive milk-protein-derived peptides were incubated with mid-log cells of LB and ST, which are required for yogurt production. Incubations were performed at pH 4.5 and 7.0, and samples removed at various time points were analysed by reversed-phase high-performance liquid chromatography (RP-HPLC). The peptides remained mostly intact at pH 4.5 in the presence of ST strains and moderately digested by exposure to LB cells. Peptide loss occurred more rapidly and was more extensive after incubation at pH 7.0. The 11mer and 12mer bioactive peptides may be added at the end of the yogurt-making process when the pH level has dropped to 4.5, limiting the overall extent of proteolysis. The results show the feasibility of using milk-protein-based antimicrobial and hypotensive peptides as food supplements to improve the health-promoting qualities of liquid and semi-solid dairy foods prepared by the yogurt fermentation process.

Evolution of Incubation Models: Evidence from the Italian Incubation Industry

ERIC Educational Resources Information Center

Grandi, Alessandro; Grimaldi, Rosa

2004-01-01

This paper addresses the role of incubators in supporting new venture creation. A mapping of four different types of incubator is proposed: corporate private incubators (CPIs), independent private incubators (IPIs), business innovation centres (BICs) and university business incubators (UBIs). This mapping is exemplified through case studies of one…

Vaginal mucus from ewes treated with progestogen sponges affects quality of ram spermatozoa.

PubMed

Manes, Jorgelina; Ríos, Glenda; Fiorentino, María Andrea; Ungerfeld, Rodolfo

2016-03-15

The use of intravaginal sponges (IS) to synchronize estrous onset in ewes provokes vaginitis, an increase in the vaginal bacterial load, and growth of bacterial species that are not present during spontaneous estrous behavior. The objective of the study was to compare the functional sperm parameters after incubating it with mucus collected from the vagina of ewes during spontaneous estrus or estrous synchronized with IS. Pooled spermatozoa were co-incubated with: (1) vaginal mucus collected from ewes in spontaneous estrus; (2) vaginal mucus collected from ewes in estrus pretreated with progestogen-impregnated IS; (3) synthetic mucus; and (4) medium without mucus as a control group. Sperm samples were evaluated after incubating it for 30 and 90 minutes. The number of colony-forming units (CFUs/mL), pH, and osmolality were greater in the mucus collected from ewes treated with IS than from those untreated (P = 0.046; P < 0.0001, and P < 0.0001, respectively). The percentage of sperm with progressive motility was lower after incubation with vaginal mucus collected from estrous ewes treated with IS than in the other three treatments both, 30 and 90 minutes after incubation (P = 0.0009 and P < 0.0001, respectively). The sample incubated for 30 minutes with mucus from ewes treated with IS had a lower percentage of sperm with intact plasma membrane than all the other treatments (P < 0.0001). The percentage of sperm with functional membrane was significantly lower in the sample incubated for 30 minutes with vaginal mucus from ewes treated with IS than in the other three treatments (P < 0.0001). After 90 minutes, the percentage was still lower than that in the sample collected from ewes during their spontaneous estrus (P = 0.0005). The lowest percentages of sperm with acrosome damage were observed in sperm incubated with mucus collected from sheep in spontaneous estrus for 30 and 90 minutes (P < 0.0001 and P = 0.008, respectively). The percentage of apoptotic spermatozoa was greater in samples incubated during 30 minutes with vaginal mucus collected from ewes treated with IS than in the other three groups (P = 0.0005). The functionality and the viability of ram sperm is negatively affected by the cervical mucus of ewes pretreated with progestagen-impregnated IS used in estrous synchronization treatments. This may partially explain the decrease in conception rate obtained with treatments with IS. Copyright © 2016 Elsevier Inc. All rights reserved.

Evaluation of fecal culture and fecal RT-PCR to detect Mycobacterium avium ssp. paratuberculosis fecal shedding in dairy goats and dairy sheep using latent class Bayesian modeling.

PubMed

Bauman, Cathy A; Jones-Bitton, Andria; Jansen, Jocelyn; Kelton, David; Menzies, Paula

2016-09-20

The study's objective was to evaluate the ability of fecal culture (FCUL) and fecal PCR (FPCR) to identify dairy goat and dairy sheep shedding Mycobacterium avium ssp. paratuberculosis. A cross-sectional study of the small ruminant populations was performed in Ontario, Canada between October 2010 and August 2011. Twenty-nine dairy goat herds and 21 dairy sheep flocks were visited, and 20 lactating females > two years of age were randomly selected from each farm resulting in 580 goats and 397 sheep participating in the study. Feces were collected per rectum and cultured using the BD BACTEC™ MGIT™ 960 system using a standard (49 days) and an extended (240 days) incubation time, and underwent RT-PCR based on the hsp-X gene (Tetracore®). Statistical analysis was performed using a 2-test latent class Bayesian hierarchical model for each species fitted in WinBUGS. Extending the fecal culture incubation time statistically improved FCUL sensitivity from 23.1 % (95 % PI: 15.9-34.1) to 42.7 % (95 % PI: 33.0-54.5) in dairy goats and from 5.8 % (95 % PI: 2.3-12.4) to 19.0 % (95 % PI: 11.9-28.9) in dairy sheep. FPCR demonstrated statistically higher sensitivity than FCUL (49 day incubation) with a sensitivity of 31.9 % (95 % PI: 22.4-43.1) in goats and 42.6 % (95 % PI: 28.8-63.3) in sheep. Fecal culture demonstrates such low sensitivity at the standard incubation time it cannot be recommended as a screening test to detect shedding of MAP in either goats or sheep. Extending the incubation time resulted in improved sensitivity; however, it is still disappointingly low for screening purposes. Fecal PCR should be the screening test of choice in both species; however, it is important to recognize that control programs should not be based on testing alone when they demonstrate such low sensitivity.

Biotransformation of ent-kaur-16-en-19-oic acid by Psilocybe cubensis.

PubMed

Pechwang, Jaraslak; Sihanonth, Prakitsin; Pornpakakul, Surachai; Muangsin, Nongnuj; Piapukiew, Jittra; Vangnai, Alisa; Chaichit, Narongsak; Chuchawankul, Siriporn; Petsom, Amorn

2010-06-01

Biotranformation of ent-kaur-16-en-19-oic acid (1) using Psilocybe cubensis resulted in hydroxylated products. After two days of incubation, ent-16beta,17-dihydroxy-kauran-19-oic acid (2) was isolated. After further incubation for nine days, two novel metabolites, ent-12alpha,16beta,17-trihydroxy-kauran-19-oic acid (3) and ent-11alpha,16beta,17-trihydroxy-kauran-19-oic acid (4), were obtained. The metabolites were identified by spectroscopic methods and X-ray crystallography. Compounds 1-4 were evaluated for their cytotoxic properties against the human leukaemia K562 cell line; only compound 1 showed moderate activity.

Rapid-Viability PCR Method for Detection of Live, Virulent Bacillus anthracis in Environmental Samples ▿

PubMed Central

Létant, Sonia E.; Murphy, Gloria A.; Alfaro, Teneile M.; Avila, Julie R.; Kane, Staci R.; Raber, Ellen; Bunt, Thomas M.; Shah, Sanjiv R.

2011-01-01

In the event of a biothreat agent release, hundreds of samples would need to be rapidly processed to characterize the extent of contamination and determine the efficacy of remediation activities. Current biological agent identification and viability determination methods are both labor- and time-intensive such that turnaround time for confirmed results is typically several days. In order to alleviate this issue, automated, high-throughput sample processing methods were developed in which real-time PCR analysis is conducted on samples before and after incubation. The method, referred to as rapid-viability (RV)-PCR, uses the change in cycle threshold after incubation to detect the presence of live organisms. In this article, we report a novel RV-PCR method for detection of live, virulent Bacillus anthracis, in which the incubation time was reduced from 14 h to 9 h, bringing the total turnaround time for results below 15 h. The method incorporates a magnetic bead-based DNA extraction and purification step prior to PCR analysis, as well as specific real-time PCR assays for the B. anthracis chromosome and pXO1 and pXO2 plasmids. A single laboratory verification of the optimized method applied to the detection of virulent B. anthracis in environmental samples was conducted and showed a detection level of 10 to 99 CFU/sample with both manual and automated RV-PCR methods in the presence of various challenges. Experiments exploring the relationship between the incubation time and the limit of detection suggest that the method could be further shortened by an additional 2 to 3 h for relatively clean samples. PMID:21764960

Proteolysis of serum amyloid A and AA amyloid proteins by cysteine proteases: cathepsin B generates AA amyloid proteins and cathepsin L may prevent their formation

PubMed Central

Rocken, C; Menard, R; Buhling, F; Vockler, S; Raynes, J; Stix, B; Kruger, S; Roessner, A; Kahne, T

2005-01-01

Background: AA amyloidosis develops in patients with chronic inflammatory diseases. The AA amyloid proteins are proteolytic fragments obtained from serum amyloid A (SAA). Previous studies have provided evidence that endosomes or lysosomes might be involved in the processing of SAA, and contribute to the pathology of AA amyloidosis. Objective: To investigate the anatomical distribution of cathepsin (Cath) B and CathL in AA amyloidosis and their ability to process SAA and AA amyloid proteins. Methods and results: CathB and CathL were found immunohistochemically in every patient with AA amyloidosis and displayed a spatial relationship with amyloid in all the cases studied. Both degraded SAA and AA amyloid proteins in vitro. With the help of mass spectrometry 27 fragments were identified after incubation of SAA with CathB, nine of which resembled AA amyloid proteins, and seven fragments after incubation with CathL. CathL did not generate AA amyloid-like peptides. When native human AA amyloid proteins were used as a substrate 26 fragments were identified after incubation with CathB and 18 after incubation with CathL. Conclusion: The two most abundant and ubiquitously expressed lysosomal proteases can cleave SAA and AA amyloid proteins. CathB generates nine AA amyloid-like proteins by its carboxypeptidase activity, whereas CathL may prevent the formation of AA amyloid proteins by endoproteolytic activity within the N-terminal region of SAA. This is particularly interesting, because AA amyloidosis is a systemic disease affecting many organs and tissue types, almost all of which express CathB and CathL. PMID:15897303

The role of estrogen in turtle sex determination and the effect of PCBs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Crews, D.; Bergeron, J.M.; McLachlan, J.A.

1995-10-01

Gonadal sex is fixed at fertilization by specific chromosomes, a process known as genotypic sex determination (GSD). Only after the gonad is formed do hormones begin to exert an influence that modifies specific structures that eventually will differ between the sexes. Many egg-laying reptiles do not exhibit GSD but rather depend on the temperature of the incubating egg to determine the gonadal sex of the offspring, a process termed temperature-dependent sex determination (TSD). Research on TSD indicates that gonadal sex is not irrevocably set by the genetic composition inherited at fertilization but depends ultimately on which genes encoding for steroidogenicmore » enzymes and hormone receptors are activated during the midtrimester of embryonic development by temperature. Incubation temperature modifies the activity as well as the temporal and spatial sequence of enzymes and hormone receptors to determine gonad type. Estrogen is the physiologic equivalent of incubation temperature and the proximate cue that initiates female sex determination. increasing evidence indicates some polychlorinated biphenyl (PCB) compounds are capable of disrupting reproductive and endocrine function in fish, birds, and mammals, including humans. Reproductive disorders resulting from exposure to these xenobiotic compounds may include reductions in fertility, hatch rate in fish and birds, and viability of offspring, as well as alterations in hormone levels or adult sexual behaviors. Research on the mechanism through which these compounds may be acting to alter reproductive function indicates estrogenic activity, by which the compounds may be altering sexual differentiation. In TSD turtles, the estrogenic effect of some PCBs reverses gonadal sex in individuals incubating at an otherwise male-producing temperature. Furthermore, certain PCBs are synergistic in their effect at very low concentrations. 19 refs., 3 figs., 1 tab.« less

Release of an endogenous pyrogen in vitro from rabbit mononuclear cells.

PubMed

Atkins, E; Bodel, P; Francis, L

1967-08-01

The capacity of rabbit mononuclear cells to release an endogenous pyrogen (EP) in vitro has been studied. After incubation with tuberculin, preparations of predominantly monocytic cells, derived from the respiratory passages of the lungs of rabbits sensitized with BCG, were activated to release EP. Pyrogen production occurred more slowly with lung monocytes than with blood leukocytes of similarly sensitized rabbits and 9 to 10 hr incubation in a fully supportive medium was required to produce clear-cut results. As previously reported with blood leukocytes, mononuclear cells from the lungs of normal animals were also activated by tuberculin but to a lesser degree than were those from specifically sensitized rabbits. Under a variety of conditions, mononuclear cells from either spleen or lymph nodes of the same sensitized rabbits failed to release detectable amounts of pyrogen when incubated with tuberculin in vitro but were activated in a majority of instances when phagocytosis of heat-killed staphylococci was used as the stimulus. Release of pyrogen from lung monocytes appears to be an active process that is both temperature-dependent and requires protein synthesis. Neither serum antibody nor complement appears to play a role in this process. Evidence is presented that the granulocyte is the main source of pyrogen evolved by blood leukocytes incubated in vitro with OT or heat-killed staphylococci, whereas the lung macrophage and/or monocyte is responsible for most of the pyrogen released from the lung cell preparations. From these studies, it is concluded that mononuclear cells can be activated in vitro by several microbial stimuli and must be considered an additional cellular source of EP. The clinical implications of these findings for the pathogenesis of fever in granulomatous diseases where the monocyte is the predominant cell are discussed.

Evidence of biogenic corrosion of titanium after exposure to a continuous culture of thiobacillus ferrooxidans grown in thiosulfate medium

DOE Office of Scientific and Technical Information (OSTI.GOV)

Horn, J M; Martin, S I; Masterson, B

2000-12-07

Experiments were undertaken to evaluate extreme conditions under which candidate materials intended for use in a proposed nuclear waste repository might be susceptible to corrosion by endogenous microorganisms. Thiobucillus ferrooxidans, a sulfur-oxidizing bacterium, was grown in continuous culture using thiosulfate as an energy source; thiosulfate is oxidized to sulfate as a metabolic endproduct by this organism. Culture conditions were optimized to produce a high-density, metabolically active culture throughout a period of long term incubation in the presence of Alloy 22 (a high nickel-based alloy) and Titanium grade 7 (Tigr7) material coupons. After seven months incubation under these conditions, material couponsmore » were withdrawn and analyzed by high resolution microscopy and energy dispersive x-ray analyses. Alloy 22 coupons showed no detectable signs of corrosion. Tigr7, however, demonstrated distinct roughening of the coupon surface, and [presumably solubilized and precipitated] titanium was detected on Alloy 22 coupons incubated in the same T. ferrooxiduns culture vessel. Control coupons of these materials incubated in sterile thiosulfate medium did not demonstrate any signs of corrosion, thus showing that observed corrosive effects were due to the T. ferrooxidans metabolic activities. T. ferrooxidans intermediates of thiosulfate oxidation or sulfate may have caused the corrosive effects observed on Tigr7.« less

Probiotic Bacteria and their Supernatants Protect Enterocyte Cell Lines from Enteroinvasive Escherichia coli (EIEC) Invasion

PubMed Central

Khodaii, Zohreh; Ghaderian, Sayyed Mohammad Hossein; Natanzi, Mahboobeh Mehrabani

2017-01-01

Probiotic microorganisms have attracted a growing interest for prevention and therapy of gastrointestinal disorders. Many probiotic strains have been shown to inhibit growth and metabolic activity of enteropathogenic bacteria as well as their adhesion and invasion to intestinal cells. In the present study, we evaluated the interference of bacteria-free supernatants (BFS) of cultures belonging to sixteen strains of lactobacilli and bifidobacteria, with invasion of enteroinvasive Escherichia coli (EIEC) strain, using human colonic adenocarcinoma cell lines, T84 and Caco2 cells. To assess invasion of Caco-2 and T84 cells by EIEC, and measure the number of pathogens inside the enterocytes, the gentamicin protection assay was conducted. In addition, three different invasion inhibition assays were designed; namely co-incubation, pre-incubation and treatment with the BFS of probiotics. Data obtained and theoretical calculation showed that the most effective assay in the prevention of pathogen invasion was treatment with BFS. Besides, co-incubation assay was more valid than pre-incubation assay in invasion prevention. The obtained results suggest that probiotics may produce some metabolites that strongly prevent invasion of enteroinvasive E.coli into the small and large intestine. Also, probiotics are able to compete with or exclude pathogen invasion. PMID:29682490

Internalization of the PDZ and its photodynamic effect on the growth of ATCC and clinical strains of E. coli and S. aureus

NASA Astrophysics Data System (ADS)

Rodrigues da Silva, Gislene; Henrique Correia Pereira, André; Guerra Pinto, Juliana; José Raniero, Leandro; Ferreira-Strixino, Juliana

2016-09-01

The treatment of bacterial infections has been a challenge after the end of the ‘era of antibiotics’. Bacteria are capable of causing many infectious diseases; therefore, with the increasing number of bacteria becoming resistant, development of alternative therapies is needed to minimize, or even eliminate the use of antibiotics. Photodynamic therapy (PDT) is a promising alternative to fight microorganism. In view of the increasing emergence of resistant bacteria and the limitations of conventional treatment, this study evaluated the effect of photodynamic therapy with photodithazine (PDZ) in inactivating bacterial strains of E. coli and S. aureus in vitro, comparing the behavior of clinical and ATCC strains. Confocal microscopy analysis was performed to determine the internalization of the PS and spectrophotometric technique was used to determine the growth of bacteria in vitro. PDT using PDZ was able to reduce the growth of S. aureus strains using the incubation time of 24 h, whereas no satisfactory results were obtained with 15 min incubation. The E. coli strains, tested at two incubation times, did not affectively reduce bacterial growth. Therefore, it is concluded that PDT using PDZ is viable when applied to the S. aureus strains, when suitable incubation times are used.

Important factors for cell-membrane permeabilization by gold nanoparticles activated by nanosecond-laser irradiation

PubMed Central

Yao, Cuiping; Rudnitzki, Florian; Hüttmann, Gereon; Zhang, Zhenxi; Rahmanzadeh, Ramtin

2017-01-01

Purpose Pulsed-laser irradiation of light-absorbing gold nanoparticles (AuNPs) attached to cells transiently increases cell membrane permeability for targeted molecule delivery. Here, we targeted EGFR on the ovarian carcinoma cell line OVCAR-3 with AuNPs. In order to optimize membrane permeability and to demonstrate molecule delivery into adherent OVCAR-3 cells, we systematically investigated different experimental conditions. Materials and methods AuNPs (30 nm) were functionalized by conjugation of the antibody cetuximab against EGFR. Selective binding of the particles was demonstrated by silver staining, multiphoton imaging, and fluorescence-lifetime imaging. After laser irradiation, membrane permeability of OVCAR-3 cells was studied under different conditions of AuNP concentration, cell-incubation medium, and cell–AuNP incubation time. Membrane permeability and cell viability were evaluated by flow cytometry, measuring propidium iodide and fluorescein isothiocyanate–dextran uptake. Results Adherently growing OVCAR-3 cells can be effectively targeted with EGFR-AuNP. Laser irradiation led to successful permeabilization, and 150 kDa dextran was successfully delivered into cells with about 70% efficiency. Conclusion Antibody-targeted and laser-irradiated AuNPs can be used to deliver molecules into adherent cells. Efficacy depends not only on laser parameters but also on AuNP:cell ratio, cell-incubation medium, and cell–AuNP incubation time. PMID:28848345

Randomized Vehicle-Controlled Study of Short Drug Incubation Aminolevulinic Acid Photodynamic Therapy for Actinic Keratoses of the Face or Scalp.

PubMed

Pariser, David M; Houlihan, Anna; Ferdon, Mary Beth; Berg, James E

2016-03-01

Aminolevulinic acid photodynamic therapy (ALA-PDT) can be effective and well tolerated when applied over a broad area and for short drug incubation times. To evaluate the effect of short-incubation time and application method on the safety and efficacy of ALA-PDT versus vehicle (VEH-PDT) in the treatment of actinic keratoses (AKs) of the face or scalp. Aminolevulinic acid or VEH was applied to face or scalp as a broad area application for 1, 2, or 3 hours or as a spot application for 2 hours before blue light activation. An identical treatment was repeated at Week 8 if any AK lesions remained. Median AK clearance rate for ALA-treated subjects ranged from 68% to 79% at Week 12, compared with 7% of the VEH-treated group (p < .0001). Complete clearance rate for ALA-treated subjects ranged from 17% (8/46) to 30% (14/47) at Week 12, compared with 2% (1/46) of the VEH-treated group (p = .0041). The safety profile seen in this study is consistent with previously reported side effects of the therapy. Short-incubation ALA-PDT was found to be superior to VEH-PDT for AK lesion clearance. A second treatment improves efficacy.

Red cell surface changes in cold agglutination

PubMed Central

Salsbury, A. J.; Clarke, J. A.; Shand, W. S.

1968-01-01

Surface changes in red blood cells undergoing cold agglutination have been investigated using the Cambridge Stereoscan electron microscope. On incubation of red cells with a cold agglutinin of anti-I specificity at 4°C, circular shadows on the red cell membrane developed within 2 min. At the same time the membrane showed a granularity and processes began to develop on the surface. These processes increased in length, the processes of contiguous cells became interlinked and agglutination was complete after incubation of 1 hr. On warming an agglutinated specimen, the process was reversed with separation of red cells and retraction of the finger-like processes to yield discrete red cells of normal appearance. The addition of heparin in vivo prevented agglutination but did not inhibit surface changes completely. Complement appeared to play no part in the production of cold agglutination due to these antibodies or in the reversal of agglutination by warming. The significance of the surface changes described in relation to previous information on the mechanism of agglutination, has been discussed. ImagesFig. 1Fig. 2Fig. 3Fig. 4Fig. 5Fig. 6Fig. 7Fig. 8Fig. 9Fig. 10Fig. 11 PMID:5655472

Assessment of hazardous wastes for genotoxicity

DOE Office of Scientific and Technical Information (OSTI.GOV)

DeMarini, D.M.; Houk, V.S.

1987-09-01

The authors have evaluated a group of short-term bioassays to identify those that may be suitable for screening large numbers of diverse hazardous industrial wastes for genotoxicity. Fifteen wastes (and dichloromethane extracts of these wastes) from a variety of manufacturing processes were tested for mutagenicity in Salmonella typhimurium strains TA98 and TA100 with and without Aroclor 1254-induced rat-liver S9. Ten of these wastes were fed by gavage to F-344 male rats, and the raw urines were assayed for mutagenicity in the presence of beta-glucuronidase in strain TA98 with S9. Six of these urines were extracted by C18/methanol elution, incubated withmore » beta-glucuronidase, and evaluated in strain TA98 with S9 and beta-glucuronidase. Fourteen of the wastes were examined for their ability to induce prophage lambda in Escherichia coli in a microsuspension assay. A second set of wastes, consisting of four industrial wastes, were evaluated in Salmonella and in a series of mammalian cell assays to measure mutagenicity, cytogenetic effects, and transformation.« less