Impact of culture conditions on the chlorophyll content of microalgae for biotechnological applications.

PubMed

da Silva Ferreira, Veronica; Sant'Anna, Celso

2017-01-01

Chlorophyll is a commercially important natural green pigment responsible for the absorption of light energy and its conversion into chemical energy via photosynthesis in plants and algae. This bioactive compound is widely used in the food, cosmetic, and pharmaceutical industries. Chlorophyll has been consumed for health benefits as a nutraceutical agent with antioxidant, anti-inflammatory, antimutagenic, and antimicrobial properties. Microalgae are photosynthesizing microorganisms which can be extracted for several high-value bioproducts in the biotechnology industry. These microorganisms are highly efficient at adapting to physicochemical variations in the local environment. This allows optimization of culture conditions for inducing microalgal growth and biomass production as well as for changing their biochemical composition. The modulation of microalgal culture under controlled conditions has been proposed to maximize chlorophyll accumulation. Strategies reported in the literature to promote the chlorophyll content in microalgae include variation in light intensity, culture agitation, and changes in temperature and nutrient availability. These factors affect chlorophyll concentration in a species-specific manner; therefore, optimization of culture conditions has become an essential requirement. This paper provides an overview of the current knowledge on the effects of key environmental factors on microalgal chlorophyll accumulation, focusing on small-scale laboratory experiments.

[Clinical governance and patient safety culture in clinical laboratories in the Spanish National Health System].

PubMed

Giménez-Marín, Á; Rivas-Ruiz, F

To conduct a situational analysis of patient safety culture in public laboratories in the Spanish National Health System and to determine the clinical governance variables that most strongly influence patient safety. A descriptive cross-sectional study was carried out, in which a Survey of Patient Safety in Clinical Laboratories was addressed to workers in 26 participating laboratories. In this survey, which consisted of 45 items grouped into 6 areas, scores were assigned on a scale from 0 to 100 (where 0 is the lowest perception of patient safety). Laboratory managers were asked specific questions about quality management systems and technology. The mean scores for the 26 participating hospitals were evaluated, and the following results observed: in 4of the 6areas, the mean score was higher than 70 points. In the third area (equipment and resources) and the fourth area (working conditions), the scores were lower than 60 points. Every hospital had a digital medical record system. This 100% level of provision was followed by that of an electronic request management system, which was implemented in 82.6% of the hospitals. The results obtained show that the culture of security is homogeneous and of high quality in health service laboratories, probably due to the steady improvement observed. However, in terms of clinical governance, there is still some way to go, as shown by the presence of weaknesses in crucial dimensions of safety culture, together with variable levels of implementation of fail-safe technologies and quality management systems. Copyright © 2017 SECA. Publicado por Elsevier España, S.L.U. All rights reserved.

Semiquantitative culture of Gardnerella vaginalis in laboratory determination of nonspecific vaginitis.

PubMed Central

Ratnam, S; Fitzgerald, B L

1983-01-01

To evaluate the usefulness of quantitative cultures of Gardnerella vaginalis in the laboratory determination of nonspecific vaginitis, the actual and relative numbers of G. vaginalis in genital cultures of a general patient population were assessed semiquantitatively, and the laboratory results were then correlated with the clinical findings. Of the 1,585 women studied, 417 (26.3%) yielded G. vaginalis in culture. Of these, only 113 (27.1%) were found to have symptoms and signs consistent with nonspecific vaginitis. G. vaginalis was obtained in pure or predominant growth from 87 of 100 consecutive cases with nonspecific vaginitis and 32 of 100 consecutive cases without the symptoms or signs of vaginitis (P less than 0.001). Hence, the positive predictive value of isolation of G. vaginalis in pure and predominant growths was determined to be 73% (87 of 119). Conversely, G. vaginalis was isolated in mixed or light growth significantly more often from asymptomatic women than from symptomatic patients, i.e., 68 versus 13 cases. Therefore, the negative predictive value of isolation of G. vaginalis in mixed and light growths was found to be 84% (68 of 81). Quantitation of the relative amount of G. vaginalis growth had higher predictive values as compared with the assessment of G. vaginalis growth alone. We conclude that quantitative culture of G. vaginalis is essential to obtain maximum reliability of culture results in the laboratory determination of nonspecific vaginitis. Although quantitated cultures of G. vaginalis have high predictive values, laboratory results must be interpreted in conjunction with the clinical findings. PMID:6604735

Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory.

PubMed

Bowey-Dellinger, Kristen; Dixon, Luke; Ackerman, Kristin; Vigueira, Cynthia; Suh, Yewseok K; Lyda, Todd; Sapp, Kelli; Grider, Michael; Crater, Dinene; Russell, Travis; Elias, Michael; Coffield, V McNeil; Segarra, Verónica A

2017-01-01

Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented-one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

42 CFR 493.1403 - Condition: Laboratories performing moderate complexity testing; laboratory director.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Laboratories performing moderate complexity testing; laboratory director. 493.1403 Section 493.1403 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION...

42 CFR 493.1403 - Condition: Laboratories performing moderate complexity testing; laboratory director.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing moderate complexity testing; laboratory director. 493.1403 Section 493.1403 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION...

Interspecific competition and allelopathic interaction between Karenia mikimotoi and Dunaliella salina in laboratory culture

NASA Astrophysics Data System (ADS)

He, Dong; Liu, Jiao; Hao, Qiang; Ran, Lihua; Zhou, Bin; Tang, Xuexi

2016-03-01

Algal allelopathy is a manifold ecological/physiological phenomenon that is focused on chemical interactions and autotoxicity. We investigated the allelopathic interactions between Karenia mikimotoi and Dunaliella salina in laboratory cultures based on diff erent temperature (15°C, 20°C, and 25°C) and lighting (40, 80, and 160 μmol/(m2·s)) conditions. The growth of D. salina in bi-algae culture (1:1 size/density) was significantly restrained. The results of cell-free filtrate culture indicate that direct cell-tocell contact was not necessary in interspecific competition. Further experimental results demonstrated that allelochemicals released from K. mikimotoi were markedly influenced by both temperature ( P =0.013) and irradiance ( P =0.003), resulting in diff erent growth characteristics of D. salina in filtrate mediums. Compared with the plateau period, K. mikimotoi exudates in the exponential phase had a stronger short-term inhibition effect on D. salina in normal conditions. A clear concentration-dependent relationship was observed in the effect of allelochemicals released from K. mikimotoi with low-promoting and high-repressing effects on D. Salina in a short time-scale. In addition, allelopathic substances remain stable and effective under high temperature and pressure stress. Many flocculent sediments adhering with D. salina cells were observed in all filtrate mediums, while the quantity and color depended on the original culture conditions.

Multicellular microorganisms: laboratory versus nature.

PubMed

Palková, Zdena

2004-05-01

Our present in-depth knowledge of the physiology and regulatory mechanisms of microorganisms has arisen from our ability to remove them from their natural, complex ecosystems into pure liquid cultures. These cultures are grown under optimized laboratory conditions and allow us to study microorganisms as individuals. However, microorganisms naturally grow in conditions that are far from optimal, which causes them to become organized into multicellular communities that are better protected against the harmful environment. Moreover, this multicellular existence allows individual cells to differentiate and acquire specific properties, such as forming resistant spores, which benefit the whole population. The relocation of natural microorganisms to the laboratory can result in their adaptation to these favourable conditions, which is accompanied by complex changes that include the repression of some protective mechanisms that are essential in nature. Laboratory microorganisms that have been cultured for long periods under optimized conditions might therefore differ markedly from those that exist in natural ecosystems.

Tetrahymena in the laboratory: strain resources, methods for culture, maintenance, and storage.

PubMed

Cassidy-Hanley, Donna M

2012-01-01

The ciliated protozoan Tetrahymena thermophila has been an important model system for biological research for many years. During that time, a variety of useful strains, including highly inbred stocks, a collection of diverse mutant strains, and wild cultivars from a variety of geographical locations have been identified. In addition, thanks to the efforts of many different laboratories, optimal conditions for growth, maintenance, and storage of Tetrahymena have been worked out. To facilitate the efficient use of Tetrahymena, especially by those new to the system, this chapter presents a brief description of many available Tetrahymena strains and lists possible resources for obtaining viable cultures of T. thermophila and other Tetrahymena species. Descriptions of commonly used media, methods for cell culture and maintenance, and protocols for short- and long-term storage are also presented. Copyright © 2012 Elsevier Inc. All rights reserved.

Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory †

PubMed Central

Bowey-Dellinger, Kristen; Dixon, Luke; Ackerman, Kristin; Vigueira, Cynthia; Suh, Yewseok K.; Lyda, Todd; Sapp, Kelli; Grider, Michael; Crater, Dinene; Russell, Travis; Elias, Michael; Coffield, V. McNeil; Segarra, Verónica A.

2017-01-01

Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented—one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research. PMID:28861134

Optimization to the Culture Conditions for Phellinus Production with Regression Analysis and Gene-Set Based Genetic Algorithm

PubMed Central

Li, Zhongwei; Xin, Yuezhen; Wang, Xun; Sun, Beibei; Xia, Shengyu; Li, Hui

2016-01-01

Phellinus is a kind of fungus and is known as one of the elemental components in drugs to avoid cancers. With the purpose of finding optimized culture conditions for Phellinus production in the laboratory, plenty of experiments focusing on single factor were operated and large scale of experimental data were generated. In this work, we use the data collected from experiments for regression analysis, and then a mathematical model of predicting Phellinus production is achieved. Subsequently, a gene-set based genetic algorithm is developed to optimize the values of parameters involved in culture conditions, including inoculum size, PH value, initial liquid volume, temperature, seed age, fermentation time, and rotation speed. These optimized values of the parameters have accordance with biological experimental results, which indicate that our method has a good predictability for culture conditions optimization. PMID:27610365

Conditioning laboratory cats to handling and transport.

PubMed

Gruen, Margaret E; Thomson, Andrea E; Clary, Gillian P; Hamilton, Alexandra K; Hudson, Lola C; Meeker, Rick B; Sherman, Barbara L

2013-10-01

As research subjects, cats have contributed substantially to our understanding of biological systems, from the development of mammalian visual pathways to the pathophysiology of feline immunodeficiency virus as a model for human immunodeficiency virus. Few studies have evaluated humane methods for managing cats in laboratory animal facilities, however, in order to reduce fear responses and improve their welfare. The authors describe a behavioral protocol used in their laboratory to condition cats to handling and transport. Such behavioral conditioning benefits the welfare of the cats, the safety of animal technicians and the quality of feline research data.

Urine culture contamination: a College of American Pathologists Q-Probes study of 127 laboratories.

PubMed

Bekeris, Leonas G; Jones, Bruce Allen; Walsh, Molly K; Wagar, Elizabeth A

2008-06-01

While urine culture contamination may not be completely avoidable, some laboratories have lower contamination rates than others. A College of American Pathologists (CAP) 1998 Q-Probes study showed that many interventions commonly assumed to reduce contamination were not demonstrably effective. This article revisits the issue. To examine the frequency of urine culture contamination, review current laboratory practices in the collection of urine culture specimens, and determine practice characteristics that may be associated with the contamination rate. Laboratories participating in a CAP Q-Probes study were required to prospectively collect data on 120 consecutive urine culture specimens and provide information on the patient's demographics (age and sex), the location where the specimen was collected, how the specimen was handled, the number of isolates in quantities greater than or equal to 10,000 colony-forming units (CFU)/mL, and whether the laboratory considered the specimen to be contaminated. Specific inclusion and exclusion criteria were provided to the participants. Each laboratory completed a supplemental questionnaire that probed for specific laboratory urine culture collection practices. One hundred twenty-seven laboratories participated in the study. Results from a total of 14,739 urine specimens were received. For the purpose of this study, a urine specimen was determined to be contaminated if the culture yielded more than 2 isolates in quantities greater than or equal to 10,000 CFU/mL. Using these criteria the median institution had a contamination rate of 15.0%. Laboratories in the 10th percentile (low performance) had an average contamination rate of 41.7%, while laboratories in the 90th percentile had an average rate of 0.8%. The collection site had no influence on the contamination rate, but postcollection processing, especially refrigeration of the specimen, had a substantial effect. Providing instruction to patients produced a statistically

A multi-laboratory profile of Mycoplasma contamination in Lawsonia intracellularis cultures

PubMed Central

2012-01-01

Background During the routine laboratory cultivation of Lawsonia intracellularis, Mycoplasma contamination has been a frequent problem. When Mycoplasma contamination occurs in laboratories that study L. intracellularis, the cultures must be discarded for 4 reasons: 1) Mycoplasma is inevitably concentrated along with L. intracellularis during the passage of L. intracellularis; 2) Mycoplasma inhibits the growth of L. intracellularis; and 3) it is impossible to selectively eliminate Mycoplasma in L. intracellularis cultures. In this study, we observed the contamination of Mycoplasma species during L. intracellularis cultivation among multiple laboratories. Results The presence of a Mycoplasma infection in the L. intracellularis cultures was verified using polymerase chain reaction (PCR), and a sequence analysis of the partial 16S rRNA and 23S rRNA genes was performed. A PCR-based assay using genus-specific universal primers revealed that 29 (85.3%) of the 34 cultures were contaminated with Mycoplasma, including 26 with M. hyorhinis (89.2%), 2 with M. orale (6.9%), and 1 with M. fermentans (3.4%). The Mycoplasma contamination was not the result of infection with material of pig origin. McCoy cells, which are required for the cultivation of L. intracellularis, were also ruled out as the source of the Mycoplasma contamination. Conclusions In this study, M. hyorhinis was identified as the most common mollicute that contaminated L. intracellularis cultures. Whether L. intracellularis enhances the biological properties of Mycoplasma to promote infection in McCoy cells is not known. Because the McCoy cell line stocks that were used simultaneously were all negative for Mycoplasma, and the same worker handled both the McCoy cells to maintain the bacteria and the L. intracellularis cultures, it is possible that the L. intracellularis cultures are more vulnerable to Mycoplasma contamination. Taken together, these results suggest that continuous cultures of L. intracellularis

Identifying innovation in laboratory studies of cultural evolution: rates of retention and measures of adaptation

PubMed Central

Caldwell, Christine A.; Cornish, Hannah; Kandler, Anne

2016-01-01

In recent years, laboratory studies of cultural evolution have become increasingly prevalent as a means of identifying and understanding the effects of cultural transmission on the form and functionality of transmitted material. The datasets generated by these studies may provide insights into the conditions encouraging, or inhibiting, high rates of innovation, as well as the effect that this has on measures of adaptive cultural change. Here we review recent experimental studies of cultural evolution with a view to elucidating the role of innovation in generating observed trends. We first consider how tasks are presented to participants, and how the corresponding conceptualization of task success is likely to influence the degree of intent underlying any deviations from perfect reproduction. We then consider the measures of interest used by the researchers to track the changes that occur as a result of transmission, and how these are likely to be affected by differing rates of retention. We conclude that considering studies of cultural evolution from the perspective of innovation provides us with valuable insights that help to clarify important differences in research designs, which have implications for the likely effects of variation in retention rates on measures of cultural adaptation. PMID:26926283

Response of sago pondweed, a submerged aquatic macrophyte, to herbicides in three laboratory culture systems

USGS Publications Warehouse

Fleming, W.J.; Ailstock, M.S.; Momot, J.J.; Norman, C.M.; Gorsuch, Joseph W.; Lower, William R.; Wang, Wun-cheng; Lewis, M.A.

1991-01-01

The phytotoxicity of atrazine, paraquat, glyphosate, and alachlor to sago pondweed (Potamogeton pectinatus), a submerged aquatic macrophyte, was tested under three types of laboratory culture conditions. In each case, tests were conducted in static systems, the test period was four weeks, and herbicide exposure was chronic, resulting from a single addition of herbicide to the test vessels at the beginning of the test period. The three sets of test conditions employed were(1) axenic cultures in 125-mL flasks containing a nutrient media and sucrose; (2) a microcosm system employing 18.9-L buckets containing a sand, shell, and peat substrate; and (3) an algae-free system employing O.95-L jars containing reconstituted freshwater and a nutrient agar substrate. The primary variable measured was biomass production. Plants grew well in all three test systems, with biomass of untreated plants increasing by a factor of about 5 to 6.5 during the four-week test period. Biomass production in response to herbicide exposure differed significantly among culture systems, which demonstrates the need for a standardized testing protocol for evaluating the effects of toxics on submerged aquatic plants.

42 CFR 493.1230 - Condition: General laboratory systems.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: General laboratory systems. 493.1230 Section 493.1230 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN... overall quality of the general laboratory systems and correct identified problems as specified in § 493...

19 CFR 113.67 - Commercial gauger and commercial laboratory bond conditions.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 19 Customs Duties 1 2010-04-01 2010-04-01 false Commercial gauger and commercial laboratory bond... SECURITY; DEPARTMENT OF THE TREASURY CUSTOMS BONDS Customs Bond Conditions § 113.67 Commercial gauger and commercial laboratory bond conditions. Commercial Gauger Bond Conditions (a) Commercial gauger bond...

Clinicians' interpretations of point of care urine culture versus laboratory culture results: analysis from the four-country POETIC trial of diagnosis of uncomplicated urinary tract infection in primary care.

PubMed

Hullegie, Saskia; Wootton, Mandy; Verheij, Theo J M; Thomas-Jones, Emma; Bates, Janine; Hood, Kerenza; Gal, Micaela; Francis, Nick A; Little, Paul; Moore, Michael; Llor, Carl; Pickles, Timothy; Gillespie, David; Kirby, Nigel; Brugman, Curt; Butler, Christopher C

2017-08-01

Urine culture at the point of care minimises delay between obtaining the sample and agar inoculation in a microbiology laboratory, and quantification and sensitivity results can be available more rapidly in primary care. To identify the degree to which clinicians' interpretations of a point-of-care-test (POCT) urine culture (Flexicult™ SSI-Urinary Kit) agrees with laboratory culture in women presenting to primary care with symptoms of uncomplicated urinary tract infections (UTI). Primary care clinicians used the Flexicult™-POCT, recorded their findings and took a photograph of the result, which was interpreted by microbiology laboratory technicians. Urine samples were additionally processed in routine care laboratories. Cross tabulations were used to identify important differences in organism identification, quantification and antibiotic susceptibility between these three sources of data. The influence of various laboratory definitions for UTI on culture were assessed. Primary care clinicians identified 202/289 urine samples (69.9%) as positive for UTI using the Flexicult™-POCT, whereas laboratory culture identified 94-190 (32.5-65.7%) as positive, depending on definition thresholds. 82.9% of samples identified positive for E. coli on laboratory culture were also considered positive for E. coli using the Flexicult™ -POCT, and susceptibilities were reasonably concordant. There were major discrepancies between laboratory staff interpretation of Flexicult™ photographs, clinicians' interpretation of the Flexicult™ test, and laboratory culture results. Flexicult™-POCT overestimated the positivity rate of urine samples for UTI when laboratory culture was used as the reference standard. However, it is unclear whether point-of-care or laboratory based urine culture provides the most valid diagnostic information. © The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Rearing and Maintaining Midge Cultures (Chironomus tentans) for Laboratory Studies.

ERIC Educational Resources Information Center

Hein, John; Mahadeva, Madhu N.

1992-01-01

The life history of the Chironomus tentans can be observed in easily established and maintained laboratory cultures. Projects for the classroom include observing hydration of an egg mass; embryonic development, hatching and larval feeding; larval activity; and mating activity. (MDH)

Culture conditions affect cytotoxin production by Serratia marcescens.

PubMed

Carbonell, G V; Fonseca, B A; Figueiredo, L T; Darini, A L; Yanaguita, R M

1996-12-31

Cytotoxins have been implicated in the pathogenesis of bacterial infections. In this study, the influence of different culture conditions was evaluated on cytotoxin production of Serratia marcescens. Parameters such as culture media, incubation temperature, starting pH of culture medium, aeration, anaerobiosis, carbon sources, iron concentration in he culture media, and release of cell-bond toxin by polymyxin B were investigated. The data suggest that this cytotoxin is predominantly extracellular and is not induced by iron limitation. Aerobic culture with shaking resulted in higher cytotoxicity than static aerobic or anaerobic culture. Bacteria grown in glucose, sucrose or galactose were more cytotoxic than those grown in inositol or maltose. The culture conditions that were identified as optimal for cytotoxin production by Serratia marcescens were incubation temperature ranging from 30 to 37 degrees C, in medium adjusted pH 8.5, with shaking. This work will contribute to further studies on the identification of this cytotoxic activity.

Use of HyperCard to Simulate a Tissue Culture Laboratory.

ERIC Educational Resources Information Center

Nester, Bradley S.; Turney, Tully H.

1992-01-01

Describes the use of a Macintosh computer and HyperCard software to create an introduction to cell culture techniques that closely approximates a hands-on laboratory experiment. Highlights include data acquisition, data analysis, the generation of growth curves, and electronic modeling. (LRW)

Cumulative culture in the laboratory: methodological and theoretical challenges.

PubMed

Miton, Helena; Charbonneau, Mathieu

2018-05-30

In the last decade, cultural transmission experiments (transmission chains, replacement, closed groups and seeded groups) have become important experimental tools in investigating cultural evolution. However, these methods face important challenges, especially regarding the operationalization of theoretical claims. In this review, we focus on the study of cumulative cultural evolution, the process by which traditions are gradually modified and, for technological traditions in particular, improved upon over time. We identify several mismatches between theoretical definitions of cumulative culture and their implementation in cultural transmission experiments. We argue that observed performance increase can be the result of participants learning faster in a group context rather than effectively leading to a cumulative effect. We also show that in laboratory experiments, participants are asked to complete quite simple tasks, which can undermine the evidential value of the diagnostic criterion traditionally used for cumulative culture (i.e. that cumulative culture is a process that produces solutions that no single individual could have invented on their own). We show that the use of unidimensional metrics of cumulativeness drastically curtail the variation that may be observed, which raises specific issues in the interpretation of the experimental evidence. We suggest several solutions to these mismatches (learning times, task complexity and variation) and develop the use of design spaces in experimentally investigating old and new questions about cumulative culture. © 2018 The Author(s).

Multiweek cell culture project for use in upper-level biology laboratories.

PubMed

Marion, Rebecca E; Gardner, Grant E; Parks, Lisa D

2012-06-01

This article describes a laboratory protocol for a multiweek project piloted in a new upper-level biology laboratory (BIO 426) using cell culture techniques. Human embryonic kidney-293 cells were used, and several culture media and supplements were identified for students to design their own experiments. Treatments included amino acids, EGF, caffeine, epinephrine, heavy metals, and FBS. Students researched primary literature to determine their experimental variables, made their own solutions, and treated their cells over a period of 2 wk. Before this, a sterile technique laboratory was developed to teach students how to work with the cells and minimize contamination. Students designed their experiments, mixed their solutions, seeded their cells, and treated them with their control and experimental media. Students had the choice of manipulating a number of variables, including incubation times, exposure to treatment media, and temperature. At the end of the experiment, students observed the effects of their treatment, harvested and dyed their cells, counted relative cell numbers in control and treatment flasks, and determined the ratio of living to dead cells using a hemocytometer. At the conclusion of the experiment, students presented their findings in a poster presentation. This laboratory can be expanded or adapted to include additional cell lines and treatments. The ability to design and implement their own experiments has been shown to increase student engagement in the biology-related laboratory activities as well as develop the critical thinking skills needed for independent research.

Idaho National Laboratory Cultural Resource Management Annual Report FY 2006

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

Clayton F. Marler; Julie Braun; Hollie Gilbert

2007-04-01

The Idaho National Laboratory Site is home to vast numbers and a wide variety of important cultural resources representing at least a 13,500-year span of human occupation in the region. As a federal agency, the Department of Energy Idaho Operations Office has legal responsibility for the management and protection of those resources and has delegated these responsibilities to its primary contractor, Battelle Energy Alliance (BEA). The INL Cultural Resource Management Office, staffed by BEA professionals, is committed to maintaining a cultural resource management program that accepts these challenges in a manner reflecting the resources’ importance in local, regional, and nationalmore » history. This annual report summarizes activities performed by the INL Cultural Resource Management Office staff during Fiscal Year 2006. This work is diverse, far-reaching and though generally confined to INL cultural resource compliance, also includes a myriad of professional and voluntary community activities. This document is intended to be both informative to internal and external stakeholders, and to serve as a planning tool for future cultural resource management work to be conducted on the INL.« less

Heavy Metal Absorption Efficiency of two Species of Mosses (Physcomitrella patens and Funaria hygrometrica) Studied in Mercury Treated Culture under Laboratory Condition

NASA Astrophysics Data System (ADS)

Pradhan, Abanti; Kumari, Sony; Dash, Saktisradha; Prasad Biswal, Durga; Kishore Dash, Aditya; Panigrahi, Kishore C. S.

2017-08-01

As an important component of ecosystems, mosses have a strong influence on the cycling of water, energy and nutrient. Given their sensitivity to environmental change, mosses can be used as bioindicators of water quality, air pollution, metal accumulation and climate change. In the present study, the growth, differentiation and heavy metal (Hg) absorption of two species of mosses like Physcomitrella patens and Funariahygrometrica were studied in solid cultures under laboratory conditions. It was observed that, the number of gametophores developed from single inoculated gametophores after 45 days of growth of F. hygrometrica was 11±2.0 in control where as it has decreased at higher concentrations, 4±1.5 in 1ppm of mercury treatment. P. patens also shows a similar trend. The heavy metal uptake of both the species of mosses was studied. It was observed that Hg content in pseudo leaves of P. patens ranged from 0.98 ppm to 2.76 ppm at different Hg treatment (0.1-1 ppm), whereas in F. hygrometrica it ranged from 0.78 ppm to 2.43 ppm under the same treatment condition. Comparing between the Hg content in pseudo-leaves and rhizoids of P. patens and F. hygrometrica, it was observed that the Hg content was elevated about 60-64% in rhizoids than that of pseudo-leaves at 0.1% treatment level, whereas it was increased almost up to 50% in other treatment level.

Culture conditions tailored to the cell of origin are critical for maintaining native properties and tumorigenicity of glioma cells

PubMed Central

Ledur, Pítia F.; He, Hua; Harris, Alexandra R.; Minussi, Darlan C.; Zhou, Hai-Yan; Shaffrey, Mark E.; Asthagiri, Ashok; Lopes, Maria Beatriz S.; Schiff, David; Lu, Yi-Cheng; Mandell, James W.; Lenz, Guido; Zong, Hui

2016-01-01

Background Cell culture plays a pivotal role in cancer research. However, culture-induced changes in biological properties of tumor cells profoundly affect research reproducibility and translational potential. Establishing culture conditions tailored to the cancer cell of origin could resolve this problem. For glioma research, it has been previously shown that replacing serum with defined growth factors for neural stem cells (NSCs) greatly improved the retention of gene expression profile and tumorigenicity. However, among all molecular subtypes of glioma, our laboratory and others have previously shown that the oligodendrocyte precursor cell (OPC) rather than the NSC serves as the cell of origin for the proneural subtype, raising questions regarding the suitability of NSC-tailored media for culturing proneural glioma cells. Methods OPC-originated mouse glioma cells were cultured in conditions for normal OPCs or NSCs, respectively, for multiple passages. Gene expression profiles, morphologies, tumorigenicity, and drug responsiveness of cultured cells were examined in comparison with freshly isolated tumor cells. Results OPC media-cultured glioma cells maintained tumorigenicity, gene expression profiles, and morphologies similar to freshly isolated tumor cells. In contrast, NSC-media cultured glioma cells gradually lost their OPC features and most tumor-initiating ability and acquired heightened sensitivity to temozolomide. Conclusions To improve experimental reproducibility and translational potential of glioma research, it is important to identify the cell of origin, and subsequently apply this knowledge to establish culture conditions that allow the retention of native properties of tumor cells. PMID:27106408

Outcrossing and crossbreeding recovers deteriorated traits in laboratory cultured Steinernema carpocapsae nematodes

PubMed Central

Chaston, John M.; Dillman, Adler R.; Shapiro-Ilan, David I.; Bilgrami, Anwar L.; Gaugler, Randy; Hopper, Keith R.; Adams, Byron J.

2011-01-01

The nematode Steinernema carpocapsae infects and kills many pest insects in agroecosystems and is commonly used in biocontrol of these pests. Growth of the nematodes prior to distribution for biocontrol commonly results in deterioration of traits that are essential for nematode persistence in field applications. To better understand the mechanisms underlying trait deterioration of the efficacy of natural parasitism in entomopathogenic nematodes, we explored the maintenance of fitness related traits including reproductive capacity, heat tolerance, virulence to insects and `tail standing' (formerly called nictation) among laboratory-cultured lines derived from natural, randomly mating populations of S. carpocapsae. Laboratory cultured nematode lines with fitness-related trait values below wild-type levels regained wild-type levels of reproductive and heat tolerance traits when outcrossed with a non-deteriorated line, while virulence and `tail standing' did not deteriorate in our experiments. Crossbreeding two trait-deteriorated lines with each other also resulted in restoration of trait means to wild-type levels in most crossbred lines. Our results implicate inbreeding depression as the primary cause of trait deterioration in the laboratory cultured S. carpocapsae. We further suggest the possibility of creating inbred lines purged of deleterious alleles as founders in commercial nematode growth. PMID:21447341

Outcrossing and crossbreeding recovers deteriorated traits in laboratory cultured Steinernema carpocapsae nematodes.

PubMed

Chaston, John M; Dillman, Adler R; Shapiro-Ilan, David I; Bilgrami, Anwar L; Gaugler, Randy; Hopper, Keith R; Adams, Byron J

2011-06-01

The nematode Steinernema carpocapsae infects and kills many pest insects in agro-ecosystems and is commonly used in biocontrol of these pests. Growth of the nematodes prior to distribution for biocontrol commonly results in deterioration of traits that are essential for nematode persistence in field applications. To better understand the mechanisms underlying trait deterioration of the efficacy of natural parasitism in entomopathogenic nematodes, we explored the maintenance of fitness related traits including reproductive capacity, heat tolerance, virulence to insects and 'tail standing' (formerly called nictation) among laboratory-cultured lines derived from natural, randomly mating populations of S. carpocapsae. Laboratory cultured nematode lines with fitness-related trait values below wild-type levels regained wild-type levels of reproductive and heat tolerance traits when outcrossed with a non-deteriorated line, while virulence and 'tail standing' did not deteriorate in our experiments. Crossbreeding two trait-deteriorated lines with each other also resulted in restoration of trait means to wild-type levels in most crossbred lines. Our results implicate inbreeding depression as the primary cause of trait deterioration in the laboratory cultured S. carpocapsae. We further suggest the possibility of creating inbred lines purged of deleterious alleles as founders in commercial nematode growth. Copyright © 2011 Australian Society for Parasitology Inc. All rights reserved.

Linking non-culturable (qPCR) and culturable enterococci densities with hydrometeorological conditions

USGS Publications Warehouse

Byappanahalli, Muruleedhara N.; Whitman, Richard L.; Shively, Dawn A.; Nevers, Meredith B.

2010-01-01

Quantitative polymerase chain reaction (qPCR) measurement of enterococci has been proposed as a rapid technique for assessment of beach water quality, but the response of qPCR results to environmental conditions has not been fully explored. Culture-based E. coli and enterococci have been used in empirical predictive models to characterize their responses to environmental conditions and to increase monitoring frequency and efficiency. This approach has been attempted with qPCR results only in few studies. During the summer of 2006, water samples were collected from two southern Lake Michigan beaches and the nearby river outfall (Burns Ditch) and were analyzed for enterococci by culture-based and non-culture-based (i.e., qPCR) methods, as well as culture-based E. coli. Culturable enterococci densities (log CFU/100 ml) for the beaches were significantly correlated with enterococci qPCR cell equivalents (CE) (R = 0.650, P N = 32). Enterococci CE and CFU densities were highest in Burns Ditch relative to the beach sites; however, only CFUs were significantly higher (P R = 0.565, P N = 32). Culturable E. coli and enterococci densities were significantly correlated (R = 0.682, P N = 32). Regression analyses suggested that enterococci CFU could be predicted by lake turbidity, Burns Ditch discharge, and wind direction (adjusted R2 = 0.608); enterococci CE was best predicted by Burns Ditch discharge and log-transformed lake turbidity × wave height (adjusted R2 = 0.40). In summary, our results show that analytically, the qPCR method compares well to the non-culture-based method for measuring enterococci densities in beach water and that both these approaches can be predicted by hydrometeorological conditions. Selected predictors and model results highlight the differences between the environmental responses of the two method endpoints and the potentially high variance in qPCR results

Mycelial mass production of fungi Duddingtonia flagrans and Monacrosporium thaumasium under different culture conditions

PubMed Central

2013-01-01

Background Duddingtonia flagrans and Monacrosporium thaumasium are promising fungus species in veterinary biological control of gastrointestinal nematodes because of their production capacity of fungal structures (conidia and/or chlamydospores), growth efficiency in laboratory solid media and especially their predatory capacity. However, their large-scale production remains a challenge. This work aimed at evaluating the mycelial mass production of D. flagrans (AC001 and CG722) and M. thaumasium (NF34A) nematophagous fungi under different culture conditions. Results The results did not present significant differences (p > 0.05) in mycelia mass production between the isolates cultured under pH 4.0. Furthermore, after 168 hrs., the isolate CG722 presented a lower production of mycelial mass in medium CM (corn meal) (p < 0.05). Conclusion We therefore concluded the use of culture media SD (soy dextrose) and CG (corn grits) at pH values between 6.0 and 7.0 is suitable for high mycelial mass production of D. flagrans and M. thaumasium. PMID:23985336

Effects of hypoxia condition in embryogenic callus growth of soybean cell culture

NASA Astrophysics Data System (ADS)

Damanik, R. I.; Manurung, B. H.; Bayu, E. S.

2018-02-01

The study was performed at Tissue Culture Laboratory, Agrotechnology Department, University of Sumatera Utara, to investigate the effect of plant growth regulator (PGR) and embryogenic callus performance soybean cultivars on hypoxia condition. This research had two stages, induction of embryogenic callus and analysis metabolism of callus after hypoxic condition with T-test. The analysis was used factorial Completely Randomized Design with two factors. The first factors were cultivars of soybean (Baluran, Gepak Kuning, and Grobogan) and the second factors were combinations of PGR (5 mg/l 2,4-D + 1 mg/l BAP, 10 mg/l 2,4-D + 1.5 mg/l BAP, and 15 mg/l 2,4-D + 2 mg/l BAP). The result showed the cultivars, combination of PGR, and interaction between cultivars and PGR gave significant effect to weight callus. The result of T-test showed that in hypoxic condition, POD enzyme exercise on Gepak Kuning’s callus in 5 mg/l 2,4-D + 1 mg/l BAP was different before and after hypoxic condition.

Laboratory production of human prolactin from CHO cells adapted to serum-free suspension culture.

PubMed

Arthuso, Fernanda Santos; Bartolini, Paolo; Soares, Carlos Roberto Jorge

2012-08-01

Human prolactin (hPRL) is a polypeptide with 199 amino acids and a molecular mass of 23 kDa. Previously, a eukaryotic hPRL expression vector was used to transfect Chinese hamster ovary (CHO) cells: this work describes a fast and practical laboratory adaptation of these transfected cells, in ~40 days, to grow in suspension in serum-free medium. High cell densities of up to 4.0 × 10(6) cell/ml were obtained from spinner flask cultures and a stable and continuous production process was developed for at least 30 days. Two harvesting strategies were set up, 50 or 100 % of the total conditioned medium being collected daily and replaced by fresh culture medium. The volumetric productivity was 5-7 μg hPRL/ml, as determined directly in the collected medium via reversed-phase HPLC (RP-HPLC). A two-step process based on a cationic exchanger followed by size exclusion chromatography was applied to obtain purified hPRL from conditioned medium. Two hPRL isoforms, non-glycosylated and glycosylated, could also be separated by high-performance size-exclusion chromatography (HPSEC) and, when analyzed by RP-HPLC, HPSEC, Western blotting, and bioassay, were found to be comparable to the World Health Organization International Reference Reagent of hPRL. These results are useful for the practical scale-up to the pilot and industrial scale of a bioprocess based on CHO cell culture.

Minimizing the Workup of Blood Culture Contaminants: Implementation and Evaluation of a Laboratory-Based Algorithm

PubMed Central

Richter, S. S.; Beekmann, S. E.; Croco, J. L.; Diekema, D. J.; Koontz, F. P.; Pfaller, M. A.; Doern, G. V.

2002-01-01

An algorithm was implemented in the clinical microbiology laboratory to assess the clinical significance of organisms that are often considered contaminants (coagulase-negative staphylococci, aerobic and anaerobic diphtheroids, Micrococcus spp., Bacillus spp., and viridans group streptococci) when isolated from blood cultures. From 25 August 1999 through 30 April 2000, 12,374 blood cultures were submitted to the University of Iowa Clinical Microbiology Laboratory. Potential contaminants were recovered from 495 of 1,040 positive blood cultures. If one or more additional blood cultures were obtained within ±48 h and all were negative, the isolate was considered a contaminant. Antimicrobial susceptibility testing (AST) of these probable contaminants was not performed unless requested. If no additional blood cultures were submitted or there were additional positive blood cultures (within ±48 h), a pathology resident gathered patient clinical information and made a judgment regarding the isolate's significance. To evaluate the accuracy of these algorithm-based assignments, a nurse epidemiologist in approximately 60% of the cases performed a retrospective chart review. Agreement between the findings of the retrospective chart review and the automatic classification of the isolates with additional negative blood cultures as probable contaminants occurred among 85.8% of 225 isolates. In response to physician requests, AST had been performed on 15 of the 32 isolates with additional negative cultures considered significant by retrospective chart review. Agreement of pathology resident assignment with the retrospective chart review occurred among 74.6% of 71 isolates. The laboratory-based algorithm provided an acceptably accurate means for assessing the clinical significance of potential contaminants recovered from blood cultures. PMID:12089259

Air conditioning a vaccine laboratory. [Connaught Medical Research Laboratory, Toronto, Canada

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

Ross J.

1976-05-01

In 1974, the new Bacterial Vaccine Building of Connaught Medical Research Laboratories, Toronto, Canada, was opened to produce such vaccines as pertussis, typhoid, paratyphoids, and cholera and such toxoids as staphylococcus, diphtheria, and tetanus. It also produces other medicinal products. The layout of the complex and the air conditioning system necessary in all zones are described and schematically shown. (MCW)

Sleep and Culture in Children with Medical Conditions

PubMed Central

Koinis-Mitchell, Daphne

2010-01-01

Objectives To provide an integrative review of the existing literature on the interrelationships among sleep, culture, and medical conditions in children. Methods A comprehensive literature search was conducted using PubMed, Medline, and PsychINFO computerized databases and bibliographies of relevant articles. Results Children with chronic illnesses experience more sleep problems than healthy children. Cultural beliefs and practices are likely to impact the sleep of children with chronic illnesses. Few studies have examined cultural factors affecting the relationship between sleep and illness, but existing evidence suggests the relationship between sleep and illness is exacerbated for diverse groups. Conclusions Sleep is of critical importance to children with chronic illnesses. Cultural factors can predispose children both to sleep problems and to certain medical conditions. Additional research is needed to address the limitations of the existing literature, and to develop culturally sensitive interventions to treat sleep problems in children with chronic illnesses. PMID:20332222

Dynamics of genetic variability in Anastrepha fraterculus (Diptera: Tephritidae) during adaptation to laboratory rearing conditions.

PubMed

Parreño, María A; Scannapieco, Alejandra C; Remis, María I; Juri, Marianela; Vera, María T; Segura, Diego F; Cladera, Jorge L; Lanzavecchia, Silvia B

2014-01-01

Anastrepha fraterculus is one of the most important fruit fly plagues in the American continent and only chemical control is applied in the field to diminish its population densities. A better understanding of the genetic variability during the introduction and adaptation of wild A. fraterculus populations to laboratory conditions is required for the development of stable and vigorous experimental colonies and mass-reared strains in support of successful Sterile Insect Technique (SIT) efforts. The present study aims to analyze the dynamics of changes in genetic variability during the first six generations under artificial rearing conditions in two populations: a) a wild population recently introduced to laboratory culture, named TW and, b) a long-established control line, named CL. Results showed a declining tendency of genetic variability in TW. In CL, the relatively high values of genetic variability appear to be maintained across generations and could denote an intrinsic capacity to avoid the loss of genetic diversity in time. The impact of evolutionary forces on this species during the adaptation process as well as the best approach to choose strategies to introduce experimental and mass-reared A. fraterculus strains for SIT programs are discussed.

Dynamics of genetic variability in Anastrepha fraterculus (Diptera: Tephritidae) during adaptation to laboratory rearing conditions

PubMed Central

2014-01-01

Background Anastrepha fraterculus is one of the most important fruit fly plagues in the American continent and only chemical control is applied in the field to diminish its population densities. A better understanding of the genetic variability during the introduction and adaptation of wild A. fraterculus populations to laboratory conditions is required for the development of stable and vigorous experimental colonies and mass-reared strains in support of successful Sterile Insect Technique (SIT) efforts. Methods The present study aims to analyze the dynamics of changes in genetic variability during the first six generations under artificial rearing conditions in two populations: a) a wild population recently introduced to laboratory culture, named TW and, b) a long-established control line, named CL. Results Results showed a declining tendency of genetic variability in TW. In CL, the relatively high values of genetic variability appear to be maintained across generations and could denote an intrinsic capacity to avoid the loss of genetic diversity in time. Discussion The impact of evolutionary forces on this species during the adaptation process as well as the best approach to choose strategies to introduce experimental and mass-reared A. fraterculus strains for SIT programs are discussed. PMID:25471362

Idaho National Laboratory Cultural Resource Management Annual Report FY 2007

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

Julie Braun; Hollie Gilbert; Dino Lowrey

2008-03-01

The Idaho National Laboratory (INL) Site is home to vast numbers and a wide variety of important cultural resources representing at least a 13,500-year span of human land use in the region. As a federal agency, the Department of Energy Idaho Operations Office has legal responsibility for the management and protection of those resources and has delegated these responsibilities to its primary contractor, Battelle Energy Alliance (BEA). The BEA professional staff is committed to maintaining a cultural resource management program that accepts these challenges in a manner reflecting the resources’ importance in local, regional, and national history. This annual reportmore » summarizes activities performed by the INL Cultural Resource Management Office (CRMO) staff during fiscal year 2007. This work is diverse, far-reaching and though generally confined to INL cultural resource compliance, also includes a myriad of professional and voluntary community activities. This document is intended to be both informative to internal and external stakeholders, and to serve as a planning tool for future cultural resource management work to be conducted on the INL.« less

Culture media influenced laboratory outcomes but not neonatal birth weight in assisted reproductive technology.

PubMed

Yin, Tai-lang; Zhang, Yi; Li, Sai-jiao; Zhao, Meng; Ding, Jin-li; Xu, Wang-ming; Yang, Jing

2015-12-01

Whether the type of culture media utilized in assisted reproductive technology has impacts on laboratory outcomes and birth weight of newborns in in-vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) was investigated. A total of 673 patients undergoing IVF/ICSI and giving birth to live singletons after fresh embryo transfer on day 3 from Jan. 1, 2010 to Dec. 31, 2012 were included. Three types of culture media were used during this period: Quinn's Advantage (QA), Single Step Medium (SSM), and Continuous Single Culture medium (CSC). Fertilization rate (FR), normal fertilization rate (NFR), cleavage rate (CR), normal cleavage rate (NCR), good-quality embryo rate (GQER) and neonatal birth weight were compared using one-way ANOVA and χ (2) tests. Multiple linear regression analysis was performed to determine the impact of culture media on laboratory outcomes and birth weight. In IVF cycles, GQER was significantly decreased in SSM medium group as compared with QA or CSC media groups (63.6% vs. 69.0% in QA; vs. 71.3% in CSC, P=0.011). In ICSI cycles, FR, NFR and CR were significantly lower in CSC medium group than in other two media groups. No significant difference was observed in neonatal birthweight among the three groups (P=0.759). Multiple linear regression analyses confirmed that the type of culture medium was correlated with FR, NFR, CR and GQER, but not with neonatal birth weight. The type of culture media had potential influences on laboratory outcomes but did not exhibit an impact on the birth weight of singletons in ART.

The expression of native and cultured human retinal pigment epithelial cells grown in different culture conditions.

PubMed

Tian, J; Ishibashi, K; Honda, S; Boylan, S A; Hjelmeland, L M; Handa, J T

2005-11-01

To determine the transcriptional proximity of retinal pigment epithelium (RPE) cells grown under different culture conditions and native RPE. ARPE-19 cells were grown under five conditions in 10% CO(2): "subconfluent" in DMEM/F12+10% FBS, "confluent" in serum and serum withdrawn, and "differentiated" for 2.5 months in serum and serum withdrawn medium. Native RPE was laser microdissected. Total RNA was extracted, reverse transcribed, and radiolabelled probes were hybridised to an array containing 5,353 genes. Arrays were evaluated by hierarchical cluster analysis and significance analysis of microarrays. 78% of genes were expressed by native RPE while 45.3--47.7% were expressed by ARPE-19 cells, depending on culture condition. While the most abundant genes were expressed by native and cultured cells, significant differences in low abundance genes were seen. Hierarchical cluster analysis showed that confluent and differentiated, serum withdrawn cultures clustered closest to native RPE, and that serum segregated cultured cells from native RPE. The number of differentially expressed genes and their function, and profile of expressed and unexpressed genes, demonstrate differences between native and cultured cells. While ARPE-19 cells have significant value for studying RPE behaviour, investigators must be aware of how culture conditions can influence the mRNA phenotype of the cell.

Assessment of patient safety culture in clinical laboratories in the Spanish National Health System.

PubMed

Giménez-Marín, Angeles; Rivas-Ruiz, Francisco; García-Raja, Ana M; Venta-Obaya, Rafael; Fusté-Ventosa, Margarita; Caballé-Martín, Inmaculada; Benítez-Estevez, Alfonso; Quinteiro-García, Ana I; Bedini, José Luis; León-Justel, Antonio; Torra-Puig, Montserrat

2015-01-01

There is increasing awareness of the importance of transforming organisational culture in order to raise safety standards. This paper describes the results obtained from an evaluation of patient safety culture in a sample of clinical laboratories in public hospitals in the Spanish National Health System. A descriptive cross-sectional study was conducted among health workers employed in the clinical laboratories of 27 public hospitals in 2012. The participants were recruited by the heads of service at each of the participating centers. Stratified analyses were performed to assess the mean score, standardized to a base of 100, of the six survey factors, together with the overall patient safety score. 740 completed questionnaires were received (88% of the 840 issued). The highest standardized scores were obtained in Area 1 (individual, social and cultural) with a mean value of 77 (95%CI: 76-78), and the lowest ones, in Area 3 (equipment and resources), with a mean value of 58 (95%CI: 57-59). In all areas, a greater perception of patient safety was reported by the heads of service than by other staff. We present the first multicentre study to evaluate the culture of clinical safety in public hospital laboratories in Spain. The results obtained evidence a culture in which high regard is paid to safety, probably due to the pattern of continuous quality improvement. Nevertheless, much remains to be done, as reflected by the weaknesses detected, which identify areas and strategies for improvement.

Idaho National Laboratory Cultural Resource Management Office FY 2010 Activity Report

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

Hollie K. Gilbert; Clayton F. Marler; Christina L. Olson

2011-09-01

The Idaho National Laboratory (INL) Site is home to vast numbers and a wide variety of important cultural resources representing at least a 13,500 year span of human land use in the region. As a federal agency, the Department of Energy, Idaho Operations Office (DOE-ID) has legal responsibility for the management and protection of the resources and has contracted these responsibilities to Battelle Energy Alliance (BEA). The BEA professional staff is committed to maintaining a cultural resource management program that accepts the challenge of preserving INL cultural resources in a manner reflecting their importance in local, regional, and national history.more » This report summarizes activities performed by the INL Cultural Resource Management Office (CRMO) staff during fiscal year 2010. This work is diverse, far-reaching and though generally confined to INL cultural resource compliance, also includes a myriad of professional and voluntary community activities. This document is intended to be informative to both internal and external stakeholders and to serve as a planning tool for future INL cultural resource management work.« less

Culture conditions tailored to the cell of origin are critical for maintaining native properties and tumorigenicity of glioma cells.

PubMed

Ledur, Pítia F; Liu, Chong; He, Hua; Harris, Alexandra R; Minussi, Darlan C; Zhou, Hai-Yan; Shaffrey, Mark E; Asthagiri, Ashok; Lopes, Maria Beatriz S; Schiff, David; Lu, Yi-Cheng; Mandell, James W; Lenz, Guido; Zong, Hui

2016-10-01

Cell culture plays a pivotal role in cancer research. However, culture-induced changes in biological properties of tumor cells profoundly affect research reproducibility and translational potential. Establishing culture conditions tailored to the cancer cell of origin could resolve this problem. For glioma research, it has been previously shown that replacing serum with defined growth factors for neural stem cells (NSCs) greatly improved the retention of gene expression profile and tumorigenicity. However, among all molecular subtypes of glioma, our laboratory and others have previously shown that the oligodendrocyte precursor cell (OPC) rather than the NSC serves as the cell of origin for the proneural subtype, raising questions regarding the suitability of NSC-tailored media for culturing proneural glioma cells. OPC-originated mouse glioma cells were cultured in conditions for normal OPCs or NSCs, respectively, for multiple passages. Gene expression profiles, morphologies, tumorigenicity, and drug responsiveness of cultured cells were examined in comparison with freshly isolated tumor cells. OPC media-cultured glioma cells maintained tumorigenicity, gene expression profiles, and morphologies similar to freshly isolated tumor cells. In contrast, NSC-media cultured glioma cells gradually lost their OPC features and most tumor-initiating ability and acquired heightened sensitivity to temozolomide. To improve experimental reproducibility and translational potential of glioma research, it is important to identify the cell of origin, and subsequently apply this knowledge to establish culture conditions that allow the retention of native properties of tumor cells. © The Author(s) 2016. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Assessment of patient safety culture in clinical laboratories in the Spanish National Health System

PubMed Central

Giménez-Marín, Angeles; Rivas-Ruiz, Francisco; García-Raja, Ana M.; Venta-Obaya, Rafael; Fusté-Ventosa, Margarita; Caballé-Martín, Inmaculada; Benítez-Estevez, Alfonso; Quinteiro-García, Ana I.; Bedini, José Luis; León-Justel, Antonio; Torra-Puig, Montserrat

2015-01-01

Introduction There is increasing awareness of the importance of transforming organisational culture in order to raise safety standards. This paper describes the results obtained from an evaluation of patient safety culture in a sample of clinical laboratories in public hospitals in the Spanish National Health System. Material and methods A descriptive cross-sectional study was conducted among health workers employed in the clinical laboratories of 27 public hospitals in 2012. The participants were recruited by the heads of service at each of the participating centers. Stratified analyses were performed to assess the mean score, standardized to a base of 100, of the six survey factors, together with the overall patient safety score. Results 740 completed questionnaires were received (88% of the 840 issued). The highest standardized scores were obtained in Area 1 (individual, social and cultural) with a mean value of 77 (95%CI: 76-78), and the lowest ones, in Area 3 (equipment and resources), with a mean value of 58 (95%CI: 57-59). In all areas, a greater perception of patient safety was reported by the heads of service than by other staff. Conclusions We present the first multicentre study to evaluate the culture of clinical safety in public hospital laboratories in Spain. The results obtained evidence a culture in which high regard is paid to safety, probably due to the pattern of continuous quality improvement. Nevertheless, much remains to be done, as reflected by the weaknesses detected, which identify areas and strategies for improvement. PMID:26525595

PCR identification of bacteria in blood culture does not fit the daily workflow of a routine microbiology laboratory.

PubMed

Karumaa, Santra; Kärpänoja, Pauliina; Sarkkinen, Hannu

2012-03-01

We have evaluated the GenoType blood culture assay (Hain Lifescience, Nehren, Germany) for the identification of bacteria in 233 positive blood cultures and assessed its suitability in the workflow of a routine microbiology laboratory. In 68/233 (29.2%) samples, the culture result could not be confirmed by the GenoType assay due to a lack of primers in the test, multiple organisms in the sample, or inconsistency with respect to the identification by culture. Although the GenoType blood culture assay gives satisfactory results for bacteria for which primers are available, there are difficulties in applying the test in the routine microbiology laboratory.

Engineering large cartilage tissues using dynamic bioreactor culture at defined oxygen conditions.

PubMed

Daly, Andrew C; Sathy, Binulal N; Kelly, Daniel J

2018-01-01

Mesenchymal stem cells maintained in appropriate culture conditions are capable of producing robust cartilage tissue. However, gradients in nutrient availability that arise during three-dimensional culture can result in the development of spatially inhomogeneous cartilage tissues with core regions devoid of matrix. Previous attempts at developing dynamic culture systems to overcome these limitations have reported suppression of mesenchymal stem cell chondrogenesis compared to static conditions. We hypothesize that by modulating oxygen availability during bioreactor culture, it is possible to engineer cartilage tissues of scale. The objective of this study was to determine whether dynamic bioreactor culture, at defined oxygen conditions, could facilitate the development of large, spatially homogeneous cartilage tissues using mesenchymal stem cell laden hydrogels. A dynamic culture regime was directly compared to static conditions for its capacity to support chondrogenesis of mesenchymal stem cells in both small and large alginate hydrogels. The influence of external oxygen tension on the response to the dynamic culture conditions was explored by performing the experiment at 20% O 2 and 3% O 2 . At 20% O 2 , dynamic culture significantly suppressed chondrogenesis in engineered tissues of all sizes. In contrast, at 3% O 2 dynamic culture significantly enhanced the distribution and amount of cartilage matrix components (sulphated glycosaminoglycan and collagen II) in larger constructs compared to static conditions. Taken together, these results demonstrate that dynamic culture regimes that provide adequate nutrient availability and a low oxygen environment can be employed to engineer large homogeneous cartilage tissues. Such culture systems could facilitate the scaling up of cartilage tissue engineering strategies towards clinically relevant dimensions.

42 CFR 493.1481 - Condition: Laboratories performing high complexity testing; cytotechnologist.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Laboratories performing high complexity testing; cytotechnologist. 493.1481 Section 493.1481 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY...

42 CFR 493.1481 - Condition: Laboratories performing high complexity testing; cytotechnologist.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing high complexity testing; cytotechnologist. 493.1481 Section 493.1481 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY...

Idaho National Laboratory Cultural Resource Management Office FY 2011 Activity Report

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

Julie Braun Williams; Brenda R. Pace; Hollie K. Gilbert

The Idaho National Laboratory (INL) Site is home to vast numbers and a wide variety of important cultural resources representing at least a 13,500 year span of human land use in the region. As a federal agency, the Department of Energy, Idaho Operations Office (DOE-ID) has legal responsibility for the management and protection of the resources and has contracted these responsibilities to Battelle Energy Alliance (BEA). The BEA professional staff is committed to maintaining a cultural resource management program that accepts the challenge of preserving INL cultural resources in a manner reflecting their importance in local, regional, and national history.more » This report is intended as a stand-alone document that summarizes activities performed by the INL Cultural Resource Management Office (CRMO) staff during fiscal year 2011. This work is diverse, far-reaching and though generally confined to INL cultural resource compliance, also includes a myriad of professional and voluntary community activities. This document is intended to be informative to both internal and external stakeholders, serve as a planning tool for future INL cultural resource management work, and meet an agreed upon legal requirement.« less

Media Compositions for Three Dimensional Mammalian Tissue Growth Under Microgravity Culture Conditions

NASA Technical Reports Server (NTRS)

Goodwin, Thomas J. (Inventor)

1998-01-01

Normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural and blood tissue. The cells are grown in vitro under microgravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel.

Media Compositions for Three-Dimensional Mammalian Tissue Growth under Microgravity Culture Conditions

NASA Technical Reports Server (NTRS)

Goodwin, Thomas J. (Inventor)

1998-01-01

Normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural and blood tissue.The cells are grown in vitro under microgravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel.

42 CFR 493.807 - Condition: Reinstatement of laboratories performing nonwaived testing.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Reinstatement of laboratories performing nonwaived testing. 493.807 Section 493.807 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY REQUIREMENTS...

42 CFR 493.807 - Condition: Reinstatement of laboratories performing nonwaived testing.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Reinstatement of laboratories performing nonwaived testing. 493.807 Section 493.807 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY REQUIREMENTS...

42 CFR 493.1361 - Condition: Laboratories performing PPM procedures; testing personnel.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Laboratories performing PPM procedures; testing personnel. 493.1361 Section 493.1361 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY REQUIREMENTS...

42 CFR 493.1361 - Condition: Laboratories performing PPM procedures; testing personnel.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing PPM procedures; testing personnel. 493.1361 Section 493.1361 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY REQUIREMENTS...

Culture conditions defining glioblastoma cells behavior: what is the impact for novel discoveries?

PubMed

Ledur, Pítia Flores; Onzi, Giovana Ravizzoni; Zong, Hui; Lenz, Guido

2017-09-15

In cancer research, the use of established cell lines has gradually been replaced by primary cell cultures due to their better representation of in vivo cancer cell behaviors. However, a major challenge with primary culture involves the finding of growth conditions that minimize alterations in the biological state of the cells. To ensure reproducibility and translational potentials for research findings, culture conditions need to be chosen so that the cell population in culture best mimics tumor cells in vivo . Glioblastoma (GBM) is one of the most aggressive and heterogeneous tumor types and the GBM research field would certainly benefit from culture conditions that could maintain the original plethora of phenotype of the cells. Here, we review culture media and supplementation options for GBM cultures, the rationale behind their use, and how much those choices affect drug-screening outcomes. We provide an overview of 120 papers that use primary GBM cultures and discuss the current predominant conditions. We also show important primary research data indicating that "mis-cultured" glioma cells can acquire unnatural drug sensitivity, which would have devastating effects for clinical translations. Finally, we propose the concurrent test of four culture conditions to minimize the loss of cell coverage in culture.

Music Evolution in the Laboratory: Cultural Transmission Meets Neurophysiology.

PubMed

Lumaca, Massimo; Ravignani, Andrea; Baggio, Giosuè

2018-01-01

In recent years, there has been renewed interest in the biological and cultural evolution of music, and specifically in the role played by perceptual and cognitive factors in shaping core features of musical systems, such as melody, harmony, and rhythm. One proposal originates in the language sciences. It holds that aspects of musical systems evolve by adapting gradually, in the course of successive generations, to the structural and functional characteristics of the sensory and memory systems of learners and "users" of music. This hypothesis has found initial support in laboratory experiments on music transmission. In this article, we first review some of the most important theoretical and empirical contributions to the field of music evolution. Next, we identify a major current limitation of these studies, i.e., the lack of direct neural support for the hypothesis of cognitive adaptation. Finally, we discuss a recent experiment in which this issue was addressed by using event-related potentials (ERPs). We suggest that the introduction of neurophysiology in cultural transmission research may provide novel insights on the micro-evolutionary origins of forms of variation observed in cultural systems.

Music Evolution in the Laboratory: Cultural Transmission Meets Neurophysiology

PubMed Central

Lumaca, Massimo; Ravignani, Andrea; Baggio, Giosuè

2018-01-01

In recent years, there has been renewed interest in the biological and cultural evolution of music, and specifically in the role played by perceptual and cognitive factors in shaping core features of musical systems, such as melody, harmony, and rhythm. One proposal originates in the language sciences. It holds that aspects of musical systems evolve by adapting gradually, in the course of successive generations, to the structural and functional characteristics of the sensory and memory systems of learners and “users” of music. This hypothesis has found initial support in laboratory experiments on music transmission. In this article, we first review some of the most important theoretical and empirical contributions to the field of music evolution. Next, we identify a major current limitation of these studies, i.e., the lack of direct neural support for the hypothesis of cognitive adaptation. Finally, we discuss a recent experiment in which this issue was addressed by using event-related potentials (ERPs). We suggest that the introduction of neurophysiology in cultural transmission research may provide novel insights on the micro-evolutionary origins of forms of variation observed in cultural systems. PMID:29713263

Larva of Glyptotendipes (Glyptotendipes) glaucus (Meigen 1818) (Chironomidae, Diptera)-morphology by Scanning Electron Microscope (SEM), karyotype, and biology in laboratory conditions.

PubMed

Kownacki, Andrzej; Woznicka, Olga; Szarek-Gwiazda, Ewa; Michailova, Paraskeva

2016-09-21

Larvae belonging to the family Chironomidae are difficult to identify. The aim of the present study was to describe the larval morphology of G. (G.) glaucus with the aid of a Scanning Electron Microscope (SEM), the karyotype and biology based on materials obtained from laboratory culture. Describing the morphology of larvae, special attention was paid to rarely or never described structures like the maxilla (lacinia and maxillary palp), the long plate situated below the ventromental plate, and plate X situated between lacinia and mentum. The use of SEM allowed also to obtain better images of labrum and ventromental plate. Morphological features of this species have been supplemented by karyotype and biology of larvae in laboratory conditions. Under controlled experimental conditions we found non-synchronous development of G. (G.) glaucus larvae hatched from one egg mass reflected in different lengths of larvae and emerged imagoes.

Extension of laboratory-measured soil spectra to field conditions

NASA Technical Reports Server (NTRS)

Stoner, E. R.; Baumgardner, M. F.; Weismiller, R. A.; Biehl, L. L.; Robinson, B. F.

1982-01-01

Spectral responses of two glaciated soils, Chalmers silty clay loam and Fincastle silt loam, formed under prairie grass and forest vegetation, respectively, were measured in the laboratory under controlled moisture equilibria using an Exotech Model 20C spectroradiometer to obtain spectral data in the laboratory under artificial illumination. The same spectroradiometer was used outdoors under solar illumination to obtain spectral response from dry and moistened field plots with and without corn residue cover, representing the two different soils. Results indicate that laboratory-measured spectra of moist soil are directly proportional to the spectral response of that same field-measured moist bare soil over the 0.52 micrometer to 1.75 micrometer wavelength range. The magnitudes of difference in spectral response between identically treated Chalmers and Fincastle soils are greatest in the 0.6 micrometers to 0.8 micrometer transition region between the visible and near infrared, regardless of field condition or laboratory preparation studied.

Metabolite profiling of microfluidic cell culture conditions for droplet based screening.

PubMed

Bjork, Sara M; Sjostrom, Staffan L; Andersson-Svahn, Helene; Joensson, Haakan N

2015-07-01

We investigate the impact of droplet culture conditions on cell metabolic state by determining key metabolite concentrations in S. cerevisiae cultures in different microfluidic droplet culture formats. Control of culture conditions is critical for single cell/clone screening in droplets, such as directed evolution of yeast, as cell metabolic state directly affects production yields from cell factories. Here, we analyze glucose, pyruvate, ethanol, and glycerol, central metabolites in yeast glucose dissimilation to establish culture formats for screening of respiring as well as fermenting yeast. Metabolite profiling provides a more nuanced estimate of cell state compared to proliferation studies alone. We show that the choice of droplet incubation format impacts cell proliferation and metabolite production. The standard syringe incubation of droplets exhibited metabolite profiles similar to oxygen limited cultures, whereas the metabolite profiles of cells cultured in the alternative wide tube droplet incubation format resemble those from aerobic culture. Furthermore, we demonstrate retained droplet stability and size in the new better oxygenated droplet incubation format.

Mortality and pathology in brown bullheads Amieurus nebulosus associated with a spontaneous Edwardsiella ictaluri outbreak under tank culture conditions

USGS Publications Warehouse

Iwanowicz, L.R.; Griffin, A.R.; Cartwright, Deborah D.; Blazer, V.S.

2006-01-01

Brown bullheads Amieurus nebulosus (family Ictaluridae) are commonly used as a sentinel of environmental contamination. These fish are not generally cultured under laboratory conditions and little is known about their disease susceptibility. Here we report an outbreak of disease due to Edwardsiella ictaluri in a laboratory population of tank-reared, wild-caught brown bullheads. The isolate was positively identified as E. ictaluri using standard bacteriological substrate utilization tests and a monoclonal antibody specific for this bacterium. This pathogen causes a significant disease in channel catfish Ictalurus punctatus and is associated with disease in other ictalurid and non-ictalurid fishes. It appears that E. ictaluri is also a significant pathogen in brown bullheads and produces clinical signs and lesions similar but not identical to those observed in channel catfish. Since commercial sources of bullheads for laboratory tank studies are not available, precautions should be taken to prevent potential E. ictaluri disease outbreaks from wild-caught bullheads intended for laboratory research. ?? Inter-Research 2006.

Mortality and pathology in brown bullheads Amieurus nebulosus associated with a spontaneous Edwardsiella ictaluri outbreak under tank culture conditions.

PubMed

Iwanowicz, Luke R; Griffin, Alison R; Cartwright, Deborah D; Blazer, Vicki S

2006-06-23

Brown bullheads Amieurus nebulosus (family Ictaluridae) are commonly used as a sentinel of environmental contamination. These fish are not generally cultured under laboratory conditions and little is known about their disease susceptibility. Here we report an outbreak of disease due to Edwardsiella ictaluri in a laboratory population of tank-reared, wild-caught brown bullheads. The isolate was positively identified as E. ictaluri using standard bacteriological substrate utilization tests and a monoclonal antibody specific for this bacterium. This pathogen causes a significant disease in channel catfish Ictalurus punctatus and is associated with disease in other ictalurid and non-ictalurid fishes. It appears that E. ictaluri is also a significant pathogen in brown bullheads and produces clinical signs and lesions similar but not identical to those observed in channel catfish. Since commercial sources of bullheads for laboratory tank studies are not available, precautions should be taken to prevent potential E. ictaluri disease outbreaks from wild-caught bullheads intended for laboratory research.

Expert Assessment of Conditions for Accredited Quality Management System Functioning in Testing Laboratories

NASA Astrophysics Data System (ADS)

Mytych, Joanna; Ligarski, Mariusz J.

2018-03-01

The quality management systems compliant with the ISO 9001:2009 have been thoroughly researched and described in detail in the world literature. The accredited management systems used in the testing laboratories and compliant with the ISO/IEC 17025:2005 have been mainly described in terms of the system design and implementation. They have also been investigated from the analytical point of view. Unfortunately, a low number of studies concerned the management system functioning in the accredited testing laboratories. The aim of following study was to assess the management system functioning in the accredited testing laboratories in Poland. On 8 October 2015, 1,213 accredited testing laboratories were present in Poland. They investigated various scientific areas and substances/objects. There are more and more such laboratories that have various problems and different long-term experience when it comes to the implementation, maintenance and improvement of the management systems. The article describes the results of the conducted expert assessment (survey) carried out to examine the conditions for the functioning of a management system in an accredited laboratory. It also focuses on the characteristics of the accredited research laboratories in Poland. The authors discuss the selection of the external and internal conditions that may affect the accredited management system. They show how the experts assessing the selected conditions were chosen. The survey results are also presented.

Establishing a stem cell culture laboratory for clinical trials

PubMed Central

Sekiya, Elíseo Joji; Forte, Andresa; Kühn, Telma Ingrid Borges de Bellis; Janz, Felipe; Bydlowski, Sérgio Paulo; Alves, Adelson

2012-01-01

Adult stem/progenitor cells are found in different human tissues. An in vitro cell culture is needed for their isolation or for their expansion when they are not available in a sufficient quantity to regenerate damaged organs and tissues. The level of complexity of these new technologies requires adequate facilities, qualified personnel with experience in cell culture techniques, assessment of quality and clear protocols for cell production. The rules for the implementation of cell therapy centers involve national and international standards of good manufacturing practices. However, such standards are not uniform, reflecting the diversity of technical and scientific development. Here standards from the United States, the European Union and Brazil are analyzed. Moreover, practical solutions encountered for the implementation of a cell therapy center appropriate for the preparation and supply of cultured cells for clinical studies are described. Development stages involved the planning and preparation of the project, the construction of the facility, standardization of laboratory procedures and development of systems to prevent cross contamination. Combining the theoretical knowledge of research centers involved in the study of cells with the practical experience of blood therapy services that manage structures for cell transplantation is presented as the best potential for synergy to meet the demands to implement cell therapy centers. PMID:23049427

Sensitive and selective culture medium for detection of environmental Clostridium difficile isolates without requirement for anaerobic culture conditions.

PubMed

Cadnum, Jennifer L; Hurless, Kelly N; Deshpande, Abhishek; Nerandzic, Michelle M; Kundrapu, Sirisha; Donskey, Curtis J

2014-09-01

Effective and easy-to-use methods for detecting Clostridium difficile spore contamination would be useful for identifying environmental reservoirs and monitoring the effectiveness of room disinfection. Culture-based detection methods are sensitive for detecting C. difficile, but their utility is limited due to the requirement of anaerobic culture conditions and microbiological expertise. We developed a low-cost selective broth medium containing thioglycolic acid and l-cystine, termed C. difficile brucella broth with thioglycolic acid and l-cystine (CDBB-TC), for the detection of C. difficile from environmental specimens under aerobic culture conditions. The sensitivity and specificity of CDBB-TC (under aerobic culture conditions) were compared to those of CDBB (under anaerobic culture conditions) for the recovery of C. difficile from swabs collected from hospital room surfaces. CDBB-TC was significantly more sensitive than CDBB for recovering environmental C. difficile (36/41 [88%] versus 21/41 [51%], respectively; P = 0.006). C. difficile latex agglutination, an enzyme immunoassay for toxins A and B or glutamate dehydrogenase, and a PCR for toxin B genes were all effective as confirmatory tests. For 477 total environmental cultures, the specificity of CDBB-TC versus that of CDBB based upon false-positive yellow-color development of the medium without recovery of C. difficile was 100% (0 false-positive results) versus 96% (18 false-positive results), respectively. False-positive cultures for CDBB were attributable to the growth of anaerobic non-C. difficile organisms that did not grow in CDBB-TC. Our results suggest that CDBB-TC provides a sensitive and selective medium for the recovery of C. difficile organisms from environmental samples, without the need for anaerobic culture conditions. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

42 CFR 493.1453 - Condition: Laboratories performing high complexity testing; clinical consultant.

Code of Federal Regulations, 2010 CFR

2010-10-01

... testing; clinical consultant. 493.1453 Section 493.1453 Public Health CENTERS FOR MEDICARE & MEDICAID... Condition: Laboratories performing high complexity testing; clinical consultant. The laboratory must have a clinical consultant who meets the requirements of § 493.1455 of this subpart and provides clinical...

42 CFR 493.1415 - Condition: Laboratories performing moderate complexity testing; clinical consultant.

Code of Federal Regulations, 2010 CFR

2010-10-01

... complexity testing; clinical consultant. 493.1415 Section 493.1415 Public Health CENTERS FOR MEDICARE... § 493.1415 Condition: Laboratories performing moderate complexity testing; clinical consultant. The laboratory must have a clinical consultant who meets the qualification requirements of § 493.1417 of this...

42 CFR 493.1459 - Condition: Laboratories performing high complexity testing; general supervisor.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing high complexity testing; general supervisor. 493.1459 Section 493.1459 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY...

42 CFR 493.1453 - Condition: Laboratories performing high complexity testing; clinical consultant.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing high complexity testing; clinical consultant. 493.1453 Section 493.1453 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY...

42 CFR 493.1487 - Condition: Laboratories performing high complexity testing; testing personnel.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Laboratories performing high complexity testing; testing personnel. 493.1487 Section 493.1487 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY...

42 CFR 493.1447 - Condition: Laboratories performing high complexity testing; technical supervisor.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing high complexity testing; technical supervisor. 493.1447 Section 493.1447 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY...

42 CFR 493.1459 - Condition: Laboratories performing high complexity testing; general supervisor.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Laboratories performing high complexity testing; general supervisor. 493.1459 Section 493.1459 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY...

42 CFR 493.1487 - Condition: Laboratories performing high complexity testing; testing personnel.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing high complexity testing; testing personnel. 493.1487 Section 493.1487 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY...

42 CFR 493.1447 - Condition: Laboratories performing high complexity testing; technical supervisor.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Laboratories performing high complexity testing; technical supervisor. 493.1447 Section 493.1447 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY...

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.

Feasibility of establishing a biosafety level 3 tuberculosis culture laboratory of acceptable quality standards in a resource-limited setting: an experience from Uganda.

PubMed

Ssengooba, Willy; Gelderbloem, Sebastian J; Mboowa, Gerald; Wajja, Anne; Namaganda, Carolyn; Musoke, Philippa; Mayanja-Kizza, Harriet; Joloba, Moses Lutaakome

2015-01-15

Despite the recent innovations in tuberculosis (TB) and multi-drug resistant TB (MDR-TB) diagnosis, culture remains vital for difficult-to-diagnose patients, baseline and end-point determination for novel vaccines and drug trials. Herein, we share our experience of establishing a BSL-3 culture facility in Uganda as well as 3-years performance indicators and post-TB vaccine trials (pioneer) and funding experience of sustaining such a facility. Between September 2008 and April 2009, the laboratory was set-up with financial support from external partners. After an initial procedure validation phase in parallel with the National TB Reference Laboratory (NTRL) and legal approvals, the laboratory registered for external quality assessment (EQA) from the NTRL, WHO, National Health Laboratories Services (NHLS), and the College of American Pathologists (CAP). The laboratory also instituted a functional quality management system (QMS). Pioneer funding ended in 2012 and the laboratory remained in self-sustainability mode. The laboratory achieved internationally acceptable standards in both structural and biosafety requirements. Of the 14 patient samples analyzed in the procedural validation phase, agreement for all tests with NTRL was 90% (P <0.01). It started full operations in October 2009 performing smear microscopy, culture, identification, and drug susceptibility testing (DST). The annual culture workload was 7,636, 10,242, and 2,712 inoculations for the years 2010, 2011, and 2012, respectively. Other performance indicators of TB culture laboratories were also monitored. Scores from EQA panels included smear microscopy >80% in all years from NTRL, CAP, and NHLS, and culture was 100% for CAP panels and above regional average scores for all years with NHLS. Quarterly DST scores from WHO-EQA ranged from 78% to 100% in 2010, 80% to 100% in 2011, and 90 to 100% in 2012. From our experience, it is feasible to set-up a BSL-3 TB culture laboratory with acceptable quality

Introduction to cell culture.

PubMed

Philippeos, Christina; Hughes, Robin D; Dhawan, Anil; Mitry, Ragai R

2012-01-01

The basics of cell culture as applied to human cells are discussed. Biosafety when working with human tissue, which is often pathogenic, is important. The requirements for a tissue culture laboratory are described, particularly the range of equipment needed to carry out cell isolation, purification, and culture. Steps must be taken to maintain aseptic conditions to prevent contamination of cultures with micro-organisms. Basic cell-handling techniques are discussed, including choice of media, primary culture, and cryopreservation of cells so they can be stored for future use. Common assays which are used to determine cell viability and activity are considered.

Enrofloxacin degradation in broiler chicken manure under various laboratory conditions.

PubMed

Slana, Marko; Sollner-Dolenc, Marija

2016-03-01

The rate of degradation of enrofloxacin in broiler chicken manure has been characterized in the laboratory according to the CVMP guideline on determining the fate of veterinary medicinal products in manure. Degradation was followed in a flow-through system under aerobic and anaerobic conditions, in the dark and in the presence of light. The rate of degradation of enrofloxacin and the formation of its degradation products are dependent on laboratory conditions. A rapid degradation of enrofloxacin in the dark was noticed, where a shorter degradation half-life under aerobic (DT50 = 59.1 days), comparing to anaerobic conditions (DT50 = 88.9 days), was determined. The presence of light slowed down the enrofloxacin degradation half-life, which was significantly shorter under aerobic (DT50 = 115.0 days), comparing to anaerobic conditions (DT50 = 190.8 days). Desethylene-enrofoxacin was the only degradation product formed, its concentrations ranged from 2.5 to 14.9 %. The concentration of the degradation product was approximately 2.5-fold higher under aerobic conditions. Enrofloxacin degradation in sterile manure incubated under sterile conditions was marginal comparing to non-sterile conditions; after 120 days of incubation, approximately 80 % of enrofloxacin was still present in manure and only 1 % of desethylene-enrofloxacin was formed. The present work demonstrates that enrofloxacin degradation in chicken manure is relatively fast when incubated in the dark under aerobic conditions which is the recommended incubation system for chicken manure according to CVMP guideline.

Optimal culture conditions are critical for efficient expansion of human testicular somatic and germ cells in vitro.

PubMed

Gat, Itai; Maghen, Leila; Filice, Melissa; Wyse, Brandon; Zohni, Khaled; Jarvi, Keith; Lo, Kirk C; Gauthier Fisher, Andrée; Librach, Clifford

2017-03-01

To optimize culture conditions for human testicular somatic cells (TSCs) and spermatogonial stem cells. Basic science study. Urology clinic and stem cell research laboratory. Eight human testicular samples. Testicular tissues were processed by mechanical and enzymatic digestion. Cell suspensions were subjected to differential plating (DP) after which floating cells (representing germ cells) were removed and attached cells (representing TSCs) were cultured for 2 passages (P0-P1) in StemPro-34- or DMEM-F12-based medium. Germ cell cultures were established in both media for 12 days. TSC cultures: proliferation doubling time (PDT), fluorescence-activated cell sorting for CD90, next-generation sequencing for 89 RNA transcripts, immunocytochemistry for TSC and germ cell markers, and conditioned media analysis; germ cell cultures: number of aggregates. TSCs had significantly prolonged PDT in DMEM-F12 versus StemPro-34 (319.6 ± 275.8 h and 110.5 ± 68.3 h, respectively). The proportion of CD90-positive cells increased after P1 in StemPro-34 and DMEM-F12 (90.1 ± 10.8% and 76.5 ± 17.4%, respectively) versus after DP (66.3 ± 7%). Samples from both media after P1 clustered closely in the principle components analysis map whereas those after DP did not. After P1 in either medium, CD90-positive cells expressed TSC markers only, and fibroblast growth factor 2 and bone morphogenetic protein 4 were detected in conditioned medium. A higher number of germ cell aggregates formed in DMEM-F12 (59 ± 39 vs. 28 ± 17, respectively). Use of DMEM-F12 reduces TSC proliferation while preserving their unique characteristics, leading to improved germ cell aggregates formation compared with StemPro-34, the standard basal medium used in the majority of previous reports. Copyright © 2017. Published by Elsevier Inc.

Culturally relevant inquiry-based laboratory module implementations in upper-division genetics and cell biology teaching laboratories.

PubMed

Siritunga, Dimuth; Montero-Rojas, María; Carrero, Katherine; Toro, Gladys; Vélez, Ana; Carrero-Martínez, Franklin A

2011-01-01

Today, more minority students are entering undergraduate programs than ever before, but they earn only 6% of all science or engineering PhDs awarded in the United States. Many studies suggest that hands-on research activities enhance students' interest in pursuing a research career. In this paper, we present a model for the implementation of laboratory research in the undergraduate teaching laboratory using a culturally relevant approach to engage students. Laboratory modules were implemented in upper-division genetics and cell biology courses using cassava as the central theme. Students were asked to bring cassava samples from their respective towns, which allowed them to compare their field-collected samples against known lineages from agricultural stations at the end of the implementation. Assessment of content and learning perceptions revealed that our novel approach allowed students to learn while engaged in characterizing Puerto Rican cassava. In two semesters, based on the percentage of students who answered correctly in the premodule assessment for content knowledge, there was an overall improvement of 66% and 55% at the end in the genetics course and 24% and 15% in the cell biology course. Our proposed pedagogical model enhances students' professional competitiveness by providing students with valuable research skills as they work on a problem to which they can relate.

Culturally Relevant Inquiry-Based Laboratory Module Implementations in Upper-Division Genetics and Cell Biology Teaching Laboratories

PubMed Central

Siritunga, Dimuth; Montero-Rojas, María; Carrero, Katherine; Toro, Gladys; Vélez, Ana; Carrero-Martínez, Franklin A.

2011-01-01

Today, more minority students are entering undergraduate programs than ever before, but they earn only 6% of all science or engineering PhDs awarded in the United States. Many studies suggest that hands-on research activities enhance students’ interest in pursuing a research career. In this paper, we present a model for the implementation of laboratory research in the undergraduate teaching laboratory using a culturally relevant approach to engage students. Laboratory modules were implemented in upper-division genetics and cell biology courses using cassava as the central theme. Students were asked to bring cassava samples from their respective towns, which allowed them to compare their field-collected samples against known lineages from agricultural stations at the end of the implementation. Assessment of content and learning perceptions revealed that our novel approach allowed students to learn while engaged in characterizing Puerto Rican cassava. In two semesters, based on the percentage of students who answered correctly in the premodule assessment for content knowledge, there was an overall improvement of 66% and 55% at the end in the genetics course and 24% and 15% in the cell biology course. Our proposed pedagogical model enhances students’ professional competitiveness by providing students with valuable research skills as they work on a problem to which they can relate. PMID:21885825

[Preliminary Study of Lonicera hypoglauca on Germination Conditions of Sand Culture Seeds and Sterilization Method of Sand Culture Seedling Sterilization].

PubMed

Tan, Mu-xiu; Zeng, Wen-wen; Wei, Peng-xiao; Mo, Qiao-cheng; Pu, Zu-ning; Cen, Xiu-fen; Shi, Feng-hua

2015-05-01

To explore the germination conditions of Lonicera hypoglauca sand culture seeds and the effects of sand culture seedlings sterilization. 0.1% HgCl2 with different sterilization time, different illumination time and temperature culture condition were adopted to study the germination conditions of sand culture seeds. Different sterilization treatments and different hardening-seedling days were used to test the sterilization effect of sand culture seedlings. The sterilization effect of the combination of 75% ethanol 30 s + 0.1% HgCl2 5 min on Lonicera hypoglauca seeds was the optimum,with the average pollution rate of 15.56%, and the average germination rate reached 51.11%. The combination of varied temperature-room temperature under light for 12 h/d was the best, with the average germination rate peaked at 75.49%, and the average germination potential reached 68.36%. The treatment of detergent liquor scrub-tap water wash on the part above the hypocotyl, which was sand cultured under the opening condition and had no root, showed the best sterilization effect, with the average pollution rate was zero, and the average survival rate peaked at 100.00%. The sterilization effect of sand culture seedlings, which was disinfected after cleaning by detergent liquor scrub-tap water wash after hardening-seeding for 30 days, was the best, with the average pollution rate of 50.00%, and the average survival rate of 100.00%. The best sterilization effect is the combination of 75% ethanol 30 s + 0.1% HgCl2 5 min; Lighting for 12 h/d of varied temperature-room temperature is regarded as the optimum culture condition. The treatment of detergent liquor scrub-tap water wash treatment on the part above the hypocotyl,which is sand cultured under the opening condition and had no root, shows the best sterilization effect. For the sand culture seedlings, before inoculated in subculture medium, should be hardening-seedling for some days and sterilized after detergent liquor scrub-tap water wash.

Laboratory Approach to the Diagnosis of Culture-Negative Infective Endocarditis.

PubMed

Subedi, S; Jennings, Z; Chen, S C-A

2017-08-01

Blood-culture negative endocarditis (BCNE) accounts for up to 35% of all cases of infective endocarditis (IE) and is a serious life-threatening condition with considerable morbidity and mortality. Rapid detection and identification of the causative pathogen is essential for timely, directed therapy. Blood-culture negative endocarditis presents a diagnostic and therapeutic challenge. Causes of BCNE are varied including: treatment with antibiotic agents prior to blood culture collection; sub-optimal specimen collection; and/or infection due to fastidious (eg. nutritionally variant streptococci), intracellular (eg. Coxiella burnetii, Bartonella species) or non-culturable or difficult to culture organisms (eg. Mycobacteria, Tropheryma whipplei and fungi); as well as non-infective aetiologies. Here, we review aetiological and diagnostic approaches to BCNE including newer molecular based techniques, with a brief summary of imaging investigation and treatment principles. Copyright © 2017 Australian and New Zealand Society of Cardiac and Thoracic Surgeons (ANZSCTS) and the Cardiac Society of Australia and New Zealand (CSANZ). Published by Elsevier B.V. All rights reserved.

Laboratory experiments duplicate conditions in the Earth’s crust

USGS Publications Warehouse

Peselnick, L.; Dieterich, J.H.; Stewart, R.M.

1974-01-01

An experimental device that simulates conditions in the Earth's crust at depths of up to 30 kilometers has been constructed by geophysicists working at the U.S Geological Survey laboratories in Menlo Park, California. A high pressure "bomb" is being used to experimentally measure the velocity of seismic waves in different types of rock at various confining pressures and temperatures. The principal purpose of these measurements is to determine the elastic and non-elastic properties of rocks and minerals under conditions of high-pressure such as exist deep in the Earth's crust. 

Clinical Microbiology Laboratories' Adoption of Culture-Independent Diagnostic Tests Is a Threat to Foodborne-Disease Surveillance in the United States.

PubMed

Shea, Shari; Kubota, Kristy A; Maguire, Hugh; Gladbach, Stephen; Woron, Amy; Atkinson-Dunn, Robyn; Couturier, Marc Roger; Miller, Melissa B

2017-01-01

INTRODUCTIONIn November 2015, the Centers for Disease Control and Prevention (CDC) sent a letter to state and territorial epidemiologists, state and territorial public health laboratory directors, and state and territorial health officials. In this letter, culture-independent diagnostic tests (CIDTs) for detection of enteric pathogens were characterized as "a serious and current threat to public health surveillance, particularly for Shiga toxin-producing Escherichia coli (STEC) and Salmonella" The document says CDC and its public health partners are approaching this issue, in part, by "reviewing regulatory authority in public health agencies to require culture isolates or specimen submission if CIDTs are used." Large-scale foodborne outbreaks are a continuing threat to public health, and tracking these outbreaks is an important tool in shortening them and developing strategies to prevent them. It is clear that the use of CIDTs for enteric pathogen detection, including both antigen detection and multiplex nucleic acid amplification techniques, is becoming more widespread. Furthermore, some clinical microbiology laboratories will resist the mandate to require submission of culture isolates, since it will likely not improve patient outcomes but may add significant costs. Specimen submission would be less expensive and time-consuming for clinical laboratories; however, this approach would be burdensome for public health laboratories, since those laboratories would need to perform culture isolation prior to typing. Shari Shea and Kristy Kubota from the Association of Public Health Laboratories, along with state public health laboratory officials from Colorado, Missouri, Tennessee, and Utah, will explain the public health laboratories' perspective on why having access to isolates of enteric pathogens is essential for public health surveillance, detection, and tracking of outbreaks and offer potential workable solutions which will allow them to do this. Marc Couturier of

Longevity and survival curves of Rhinella icterica (Anura, Bufonidae) under laboratory conditions.

PubMed

Lima, M S C S; Pederassi, J; Souza, C A S

2014-05-01

Life tables and survival curves of tadpoles from Rhinella icterica species were studied in the laboratory, under abiotic conditions controlled by a purification filter, a timer and a chiller. The survival curve for larval stage confirms a great mortality trend in the initial stages, which decreases when reaching the mature morphological condition (r = -0.94). Stages 37, 38, 39, 40 and 41 showed gradual values for their age structures, while stages 42, 43 and 44 presented high variations. Based on the results under laboratory conditions, it can be concluded that the maturity of R. icterica tadpoles development between 37 and 44 stages has a negative correlation and their predicted life expectancy is a logarithmic growth curve (y=-761.96Ln(x)+5298.5).

42 CFR 493.1467 - Condition: Laboratories performing high complexity testing; cytology general supervisor.

Code of Federal Regulations, 2010 CFR

2010-10-01

... testing; cytology general supervisor. 493.1467 Section 493.1467 Public Health CENTERS FOR MEDICARE....1467 Condition: Laboratories performing high complexity testing; cytology general supervisor. For the subspecialty of cytology, the laboratory must have a general supervisor who meets the qualification...

42 CFR 493.1467 - Condition: Laboratories performing high complexity testing; cytology general supervisor.

Code of Federal Regulations, 2014 CFR

2014-10-01

... testing; cytology general supervisor. 493.1467 Section 493.1467 Public Health CENTERS FOR MEDICARE....1467 Condition: Laboratories performing high complexity testing; cytology general supervisor. For the subspecialty of cytology, the laboratory must have a general supervisor who meets the qualification...

42 CFR 493.1467 - Condition: Laboratories performing high complexity testing; cytology general supervisor.

Code of Federal Regulations, 2012 CFR

2012-10-01

... testing; cytology general supervisor. 493.1467 Section 493.1467 Public Health CENTERS FOR MEDICARE....1467 Condition: Laboratories performing high complexity testing; cytology general supervisor. For the subspecialty of cytology, the laboratory must have a general supervisor who meets the qualification...

42 CFR 493.1467 - Condition: Laboratories performing high complexity testing; cytology general supervisor.

Code of Federal Regulations, 2013 CFR

2013-10-01

... testing; cytology general supervisor. 493.1467 Section 493.1467 Public Health CENTERS FOR MEDICARE....1467 Condition: Laboratories performing high complexity testing; cytology general supervisor. For the subspecialty of cytology, the laboratory must have a general supervisor who meets the qualification...

42 CFR 493.1467 - Condition: Laboratories performing high complexity testing; cytology general supervisor.

Code of Federal Regulations, 2011 CFR

2011-10-01

... testing; cytology general supervisor. 493.1467 Section 493.1467 Public Health CENTERS FOR MEDICARE....1467 Condition: Laboratories performing high complexity testing; cytology general supervisor. For the subspecialty of cytology, the laboratory must have a general supervisor who meets the qualification...

Creating a sustainable culture of quality through the SLMTA programme in a district hospital laboratory in Kenya.

PubMed

Maruti, Phidelis M; Mulianga, Ekesa A; Wambani, Lorna N; Wafula, Melda N; Mambo, Fidelis A; Mutisya, Shadrack M; Wakaria, Eric N; Mbati, Erick M; Amayo, Angela A; Majani, Jonathan M; Nyary, Bryan; Songwe, Kilian A

2014-01-01

Bungoma District Hospital Laboratory (BDHL), which supports a 200-bed referral facility, began its Strengthening Laboratory Management Toward Accreditation (SLMTA) journey in 2011 together with eight other laboratories in the second round of SLMTA rollout in Kenya. To describe how the SLMTA programme and enhanced quality interventions changed the culture and management style at BDHL and instilled a quality system designed to sustain progress for years to come. SLMTA implementation followed the standard three-workshop series, mentorship site visits and audits. In order to build sustainability of progress, BDHL integrated quality improvement processes into its daily operations. The lab undertook a process of changing its internal culture to align all hospital stakeholders - including upper management, clinicians, laboratory staff and maintenance staff - to the mission of sustainable quality practices at BDHL. After 16 months in the SLMTA programme, BDHL improved from zero stars (38%) to four stars (89%). Over a period of two to three years, external quality assessment results improved from 47% to 87%; staff punctuality increased from 49% to 82%; clinician complaints decreased from 83% to 16; rejection rates decreased from 12% to 3%; and annual equipment repairs decreased from 40 to 15. Twelve months later the laboratory scored three stars (81%) in an external surveillance audit conducted by Kenya Accreditation Service (KENAS). Management buy-in, staff participation, use of progress-monitoring tools and feedback systems, as well as incorporation of improvement processes into routine daily activities, were vital in developing and sustaining a culture of quality improvement.

Comprehensive model of microalgae photosynthesis rate as a function of culture conditions in photobioreactors.

PubMed

Costache, T A; Acién Fernández, F Gabriel; Morales, M M; Fernández-Sevilla, J M; Stamatin, I; Molina, E

2013-09-01

In this paper, the influence of culture conditions (irradiance, temperature, pH, and dissolved oxygen) on the photosynthesis rate of Scenedesmus almeriensis cultures is analyzed. Short-run experiments were performed to study cell response to variations in culture conditions, which take place in changing environments such as outdoor photobioreactors. Experiments were performed by subjecting diluted samples of cells to different levels of irradiance, temperature, pH, and dissolved oxygen concentration. Results demonstrate the existence of photoinhibition phenomena at irradiances higher than 1,000 μE/m(2) s; in addition to reduced photosynthesis rates at inadequate temperatures or pH-the optimal values being 35 °C and 8, respectively. Moreover, photosynthesis rate reduction at dissolved oxygen concentrations above 20 mg/l is demonstrated. Data have been used to develop an integrated model based on considering the simultaneous influence of irradiance, temperature, pH, and dissolved oxygen. The model fits the experimental results in the range of culture conditions tested, and it was validated using data obtained by the simultaneous variation of two of the modified variables. Furthermore, the model fits experimental results obtained from an outdoor culture of S. almeriensis performed in an open raceway reactor. Results demonstrate that photosynthetic efficiency is modified as a function of culture conditions, and can be used to determine the proximity of culture conditions to optimal values. Optimal conditions found (T = 35 °C, pH = 8, dissolved oxygen concentration <20 mg/l) allows to maximize the use of light by the cells. The developed model is a powerful tool for the optimal design and management of microalgae-based processes, especially outdoors, where the cultures are subject to daily culture condition variations.

[Influence of liquid or solid culture conditions on the volatile components of mycelia of Isariacateinannulata].

PubMed

Zhang, Delong; Wang, Xiaodong; Lu, Ruili; Li, Kangle; Hu, Fenglin

2011-12-01

To determine the volatile components of mycelia of Isaria cateinannulata cultured under different culture conditions, and to analyze the relationships between the culture conditions and volatile metabolites. Mycelia were cultured in solid plates with SDAY medium and liquid shake flasks with SDY medium. The culture conditions were at 25 degrees C and 8 days. Volatile components in the mycelia of I. cateinannulata were extracted with simultaneous distillation extraction and analyzed by gas chromatography-mass spectrometry. Alkenes, alkanes, heterocyclic and polycyclic aromatic hydrocarbons (PAH) were existed abundantly both in the mycelia of liquid and solid cultures, but the kinds and relative concentrations of the volatile components in mycelia of liquid and solid cultures were very different. Forty-one compounds were identified from the mycelia of solid culture and 32 compounds were identified from the mycelia of liquid culture. Esters, quinones and oximes were only found in solid cultured mycelia whereas carboxylic acids were only discovered in the mycelia of liquid culture. At the same time, mycelia of liquid culture contained much more phenols. The most abundant compounds in mycelia of liquid and solid cultures were hydrocarbons. The volatile extracts of solid cultured mycelia contained 57.6% alkenes and 9.19% alkanes. The volatile extracts of liquid cultured mycelia contained 7.85% alkenes and 22.4% alkanes. Liquid or solid culture conditions influenced the volatile components of mycelia of I. cateinannulata.

Laboratory maintenance of Treponema denticola.

PubMed

Fenno, J Christopher

2005-10-01

This unit describes the methods, media, and equipment necessary for routine laboratory culture and handling of the anaerobic oral spirochete Treponema denticola. Topics discussed include nutrient requirements, recommended media formulations, and expected growth kinetics, as well as methods and equipment necessary to maintain anaerobic conditions. An additional protocol on isolation of T. denticola from clinical samples is included.

Geomagnetic storm under laboratory conditions: randomized experiment

NASA Astrophysics Data System (ADS)

Gurfinkel, Yu I.; Vasin, A. L.; Pishchalnikov, R. Yu; Sarimov, R. M.; Sasonko, M. L.; Matveeva, T. A.

2017-10-01

The influence of the previously recorded geomagnetic storm (GS) on human cardiovascular system and microcirculation has been studied under laboratory conditions. Healthy volunteers in lying position were exposed under two artificially created conditions: quiet (Q) and storm (S). The Q regime playbacks a noise-free magnetic field (MF) which is closed to the natural geomagnetic conditions on Moscow's latitude. The S regime playbacks the initially recorded 6-h geomagnetic storm which is repeated four times sequentially. The cardiovascular response to the GS impact was assessed by measuring capillary blood velocity (CBV) and blood pressure (BP) and by the analysis of the 24-h ECG recording. A storm-to-quiet ratio for the cardio intervals (CI) and the heart rate variability (HRV) was introduced in order to reveal the average over group significant differences of HRV. An individual sensitivity to the GS was estimated using the autocorrelation function analysis of the high-frequency (HF) part of the CI spectrum. The autocorrelation analysis allowed for detection a group of subjects of study which autocorrelation functions (ACF) react differently in the Q and S regimes of exposure.

Geomagnetic storm under laboratory conditions: randomized experiment.

PubMed

Gurfinkel, Yu I; Vasin, A L; Pishchalnikov, R Yu; Sarimov, R M; Sasonko, M L; Matveeva, T A

2018-04-01

The influence of the previously recorded geomagnetic storm (GS) on human cardiovascular system and microcirculation has been studied under laboratory conditions. Healthy volunteers in lying position were exposed under two artificially created conditions: quiet (Q) and storm (S). The Q regime playbacks a noise-free magnetic field (MF) which is closed to the natural geomagnetic conditions on Moscow's latitude. The S regime playbacks the initially recorded 6-h geomagnetic storm which is repeated four times sequentially. The cardiovascular response to the GS impact was assessed by measuring capillary blood velocity (CBV) and blood pressure (BP) and by the analysis of the 24-h ECG recording. A storm-to-quiet ratio for the cardio intervals (CI) and the heart rate variability (HRV) was introduced in order to reveal the average over group significant differences of HRV. An individual sensitivity to the GS was estimated using the autocorrelation function analysis of the high-frequency (HF) part of the CI spectrum. The autocorrelation analysis allowed for detection a group of subjects of study which autocorrelation functions (ACF) react differently in the Q and S regimes of exposure.

Lincoln Laboratory demonstrates highly accurate vehicle localization under adverse weather conditions

DTIC Science & Technology

2016-05-25

2016 Lincoln Laboratory demonstrates highly accurate vehicle localization under adverse weather conditions A ground-penetrating radar system...the problems limiting the development and adoption of self-driving vehicles: how can a vehicle navigate to stay within its lane when bad weather ... weather conditions, but it is challenging, even impossible, for them to work when snow covers the markings and surfaces or precipitation obscures points

Impact of culture conditions on β-carotene encapsulation using Yarrowia lipolytica cells

NASA Astrophysics Data System (ADS)

Dang, Tran Hai; Minh, Ho Thi Thu; Van Nhi, Tran Nguyen; Ngoc, Ta Thi Minh

2017-09-01

Yeast cell was reported as an effective natural preformed material for use in encapsulation of hydrophobic compounds. The encapsulation process was normally considered as passive transfer through cellular wall and cellular membrane. Beside solubility of hydrophobic compound in phospholipid membrane or plasmolysis, membrane characteristics of yeast cell which are differed between strains and influenced by culture conditions are main factors involving the accumulation of hydrophobic compound into yeast cell. In this study, the oleaginous yeast Yarrowia lipolytica was used as micro-container shell to encapsulate a high hydrophobic compound - β-carotene. Yeast cell was cultured under different conditions and wet yeast biomass was incubated with β-carotene which was dissolved in soybean oil overnight. β-carotene accumulation was then extracted and evaluated by UV-VIS spectrometry. Optimization of culture condition was investigated using the Box-Behnken model. β-carotene encapsulation efficiency in Y. lipolytica was showed to be affected by both pH of medium and agitation conditions. The highest β-carotene encapsulation efficiency was optimized at 42.8 μg/g with Y. lipolytica cultured at pH 4.5, medium volume equal to 115 ml and agitation speed at 211 rpm.

Laboratory grown subaerial biofilms on granite: application to the study of bioreceptivity.

PubMed

Vázquez-Nion, Daniel; Silva, Benita; Troiano, Federica; Prieto, Beatriz

2017-01-01

Simulated environmental colonisation of granite was induced under laboratory conditions in order to develop an experimental protocol for studying bioreceptivity. The experimental set-up proved suitable for producing subaerial biofilms by inoculating granite blocks with planktonic multi-species phototrophic cultures derived from natural biofilms. The ability of four different cultures to form biofilms was monitored over a three-month growth period via colour measurements, quantification of photosynthetic pigments and EPS, and CLSM observations. One of the cultures under study, which comprised several taxa including Bryophyta, Charophyta, Chlorophyta and Cyanobacteria, was particularly suitable as an inoculum, mainly because of its microbial richness, its rapid adaptability to the substratum and its high colonisation capacity. The use of this culture as an inoculum in the proposed experimental set-up to produce subaerial biofilms under laboratory conditions will contribute to standardising the protocols involved, thus enabling more objective assessment of the bioreceptivity of granite in further experiments.

Laboratory evaluation and application of microwave absorption properties under simulated conditions for planetary atmospheres

NASA Technical Reports Server (NTRS)

Steffes, Paul G.

1987-01-01

Laboratory measurements were conducted to evaluate properties of atmospheric gases under simulated conditions for the outer planets. A significant addition to this effort was the capability to make such measurements at millimeter wavelengths. Measurements should soon be completed on the millimeter wave absorption from ammonia under Jovian conditions. Also studied will be the feasibility of measuring the microwave and millimeter wave properties of phosphine (PH3) under simulated Jovian conditions. Further analysis and application of the laboratory results to microwave and millimeter wave absorption data for the outer planet, such as Voyager Radio Occultation experiments, will be pursued.

Chloroplast incorporation and long-term photosynthetic performance through the life cycle in laboratory cultures of Elysia timida (Sacoglossa, Heterobranchia)

PubMed Central

2014-01-01

Introduction The Mediterranean sacoglossan Elysia timida is one of the few sea slug species with the ability to sequester chloroplasts from its food algae and to subsequently store them in a functional state in the digestive gland cells for more than a month, during which time the plastids retain high photosynthetic activity (= long-term retention). Adult E. timida have been described to feed on the unicellular alga Acetabularia acetabulum in their natural environment. The suitability of E. timida as a laboratory model culture system including its food source was studied. Results In contrast to the literature reporting that juvenile E. timida feed on Cladophora dalmatica first, and later on switch to the adult diet A. acetabulum, the juveniles in this study fed directly on A. acetabulum (young, non-calcified stalks); they did not feed on the various Cladophora spp. (collected from the sea or laboratory culture) offered. This could possibly hint to cryptic speciation with no clear morphological differences, but incipient ecological differentiation. Transmission electron microscopy of chloroplasts from A. acetabulum after initial intake by juvenile E. timida showed different states of degradation — in conglomerations or singularly — and fragments of phagosome membranes, but differed from kleptoplast images of C. dalmatica in juvenile E. timida from the literature. Based on the finding that the whole life cycle of E. timida can be completed with A. acetabulum as the sole food source, a laboratory culture system was established. An experiment with PAM-fluorometry showed that cultured E. timida are also able to store chloroplasts in long-term retention from Acetabularia peniculus, which stems from the Indo-Pacific and is not abundant in the natural environment of E. timida. Variations between three experiment groups indicated potential influences of temperature on photosynthetic capacities. Conclusions E. timida is a viable laboratory model system to study

Biomass and nutrient productivities of Tetraselmis chuii under mixotrophic culture conditions with various C:N ratios

NASA Astrophysics Data System (ADS)

Lu, Lin; Wang, Jun; Yang, Guanpin; Zhu, Baohua; Pan, Kehou

2017-03-01

Mass microalgal culture plays an irreplaceable role in aquaculture, but microalgal productivity is restricted by traditional autotrophic culture conditions. In the present study, a Tetraselmis chuii strain belonging to the phylum Chlorophyta was isolated from south Yellow Sea. The growth rate and biomass productivity of this strain was higher under mixotrophic conditions with different carbon:nitrogen (C:N) ratios than those under autotrophic conditions. When the C:N ratio was 16, the optical density and biomass productivity were 3.7- and 5-fold higher than their corresponding values under autotrophic culture conditions, respectively. Moreover, T. chuii synthesized more polysaccharides and total lipids under mixotrophic conditions. In addition, T. chuii cultured under mixotrophic conditions synthesized more types of fatty acids than autotrophic culture conditions. At a C:N ratio of 16, the percentage of C16:0 and C18:1 reached 30.08% and 24.65% of the total fatty acid (TFA) content, respectively. These findings may provide a basis for large-scale mixotrophic culture of T. chuii, as a potential bait-microalga.

An intra-laboratory cultural and real-time PCR method comparison and evaluation for the detection of subclinical paratuberculosis in dairy herds.

PubMed

Heuvelink, Annet; Hassan, Abdulwahed Ahmed; van Weering, Hilmar; van Engelen, Erik; Bülte, Michael; Akineden, Ömer

2017-05-01

Mycobacterium avium subsp. paratuberculosis (MAP) is a vigorous microorganism which causes incurable chronic enteritis, Johne's disease (JD) in cattle. A target of control programmes for JD is to accurately detect MAP-infected cattle early to reduce disease transmission. The present study evaluated the efficacy of two different cultural procedures and a TaqMan real-time PCR assay for detection of subclinical paratuberculosis in dairy herds. Therefore, sixty-one faecal samples were collected from two Dutch dairy herds (n = 40 and n = 21, respectively) which were known to be MAP-ELISA positive. All individual samples were assessed using two different cultural protocols in two different laboratories. The first cultural protocol (first laboratory) included a decontamination step with 0.75% hexadecylpyridinium chloride (HPC) followed by inoculation on Herrold's egg yolk media (HEYM). The second protocol (second laboratory) comprised of a decontamination step using 4% NaOH and malachite green-oxalic acid followed by inoculation on two media, HEYM and in parallel on modified Löwenstein-Jensen media (mLJ). For the TaqMan real-time PCR assay, all faecal samples were tested in two different laboratories using TaqMan® MAP (Johne's) reagents (Life Technologies). The cultural procedures revealed positive reactions in 1.64% of the samples for cultivation protocol 1 and 6.56 and 8.20% of the samples for cultivation protocol 2, respectively. The results of the TaqMan real-time PCR performed in two different laboratories yielded 13.11 and 19.76% positive reaction. The kappa test showed proportional agreement 0.54 between the mLJ media (second laboratory) and TaqMan® real-time PCR method (second laboratory). In conclusion, the TaqMan real-time PCR could be a strongly useful and efficient assay for the detection of subclinical paratuberculosis in dairy cattle leading to an improvement in the efficiency of MAP control strategies.

Neurosphere and adherent culture conditions are equivalent for malignant glioma stem cell lines.

PubMed

Rahman, Maryam; Reyner, Karina; Deleyrolle, Loic; Millette, Sebastien; Azari, Hassan; Day, Bryan W; Stringer, Brett W; Boyd, Andrew W; Johns, Terrance G; Blot, Vincent; Duggal, Rohit; Reynolds, Brent A

2015-03-01

Certain limitations of the neurosphere assay (NSA) have resulted in a search for alternative culture techniques for brain tumor-initiating cells (TICs). Recently, reports have described growing glioblastoma (GBM) TICs as a monolayer using laminin. We performed a side-by-side analysis of the NSA and laminin (adherent) culture conditions to compare the growth and expansion of GBM TICs. GBM cells were grown using the NSA and adherent culture conditions. Comparisons were made using growth in culture, apoptosis assays, protein expression, limiting dilution clonal frequency assay, genetic affymetrix analysis, and tumorigenicity in vivo. In vitro expansion curves for the NSA and adherent culture conditions were virtually identical (P=0.24) and the clonogenic frequencies (5.2% for NSA vs. 5.0% for laminin, P=0.9) were similar as well. Likewise, markers of differentiation (glial fibrillary acidic protein and beta tubulin III) and proliferation (Ki67 and MCM2) revealed no statistical difference between the sphere and attachment methods. Several different methods were used to determine the numbers of dead or dying cells (trypan blue, DiIC, caspase-3, and annexin V) with none of the assays noting a meaningful variance between the two methods. In addition, genetic expression analysis with microarrays revealed no significant differences between the two groups. Finally, glioma cells derived from both methods of expansion formed large invasive tumors exhibiting GBM features when implanted in immune-compromised animals. A detailed functional, protein and genetic characterization of human GBM cells cultured in serum-free defined conditions demonstrated no statistically meaningful differences when grown using sphere (NSA) or adherent conditions. Hence, both methods are functionally equivalent and remain suitable options for expanding primary high-grade gliomas in tissue culture.

Neurosphere and adherent culture conditions are equivalent for malignant glioma stem cell lines

PubMed Central

Reyner, Karina; Deleyrolle, Loic; Millette, Sebastien; Azari, Hassan; Day, Bryan W.; Stringer, Brett W.; Boyd, Andrew W.; Johns, Terrance G.; Blot, Vincent; Duggal, Rohit; Reynolds, Brent A.

2015-01-01

Certain limitations of the neurosphere assay (NSA) have resulted in a search for alternative culture techniques for brain tumor-initiating cells (TICs). Recently, reports have described growing glioblastoma (GBM) TICs as a monolayer using laminin. We performed a side-by-side analysis of the NSA and laminin (adherent) culture conditions to compare the growth and expansion of GBM TICs. GBM cells were grown using the NSA and adherent culture conditions. Comparisons were made using growth in culture, apoptosis assays, protein expression, limiting dilution clonal frequency assay, genetic affymetrix analysis, and tumorigenicity in vivo. In vitro expansion curves for the NSA and adherent culture conditions were virtually identical (P=0.24) and the clonogenic frequencies (5.2% for NSA vs. 5.0% for laminin, P=0.9) were similar as well. Likewise, markers of differentiation (glial fibrillary acidic protein and beta tubulin III) and proliferation (Ki67 and MCM2) revealed no statistical difference between the sphere and attachment methods. Several different methods were used to determine the numbers of dead or dying cells (trypan blue, DiIC, caspase-3, and annexin V) with none of the assays noting a meaningful variance between the two methods. In addition, genetic expression analysis with microarrays revealed no significant differences between the two groups. Finally, glioma cells derived from both methods of expansion formed large invasive tumors exhibiting GBM features when implanted in immune-compromised animals. A detailed functional, protein and genetic characterization of human GBM cells cultured in serum-free defined conditions demonstrated no statistically meaningful differences when grown using sphere (NSA) or adherent conditions. Hence, both methods are functionally equivalent and remain suitable options for expanding primary high-grade gliomas in tissue culture. PMID:25806119

Cell culture contamination.

PubMed

Stacey, Glyn N

2011-01-01

Microbial contamination is a major issue in cell culture, but there are a range of procedures which can be adopted to prevent or eliminate contamination. Contamination may arise from the operator and the laboratory environment, from other cells used in the laboratory, and from reagents. Some infections may present a risk to laboratory workers: containment and aseptic technique are the key defence against such risks. Remedial management of suspected infection may simply mean discarding a single potentially infected culture. However, if a more widespread problem is identified, then all contaminated cultures and associated unused media that have been opened during this period should be discarded, equipment should be inspected and cleaned, cell culture operations reviewed, and isolation from other laboratories instituted until the problem is solved. Attention to training of staff, laboratory layout, appropriate use of quarantine for new cultures or cell lines, cleaning and maintenance, and quality control are important factors in preventing contamination in cell culture laboratories.

Microfluidically supported biochip design for culture of endothelial cell layers with improved perfusion conditions.

PubMed

Raasch, Martin; Rennert, Knut; Jahn, Tobias; Peters, Sven; Henkel, Thomas; Huber, Otmar; Schulz, Ingo; Becker, Holger; Lorkowski, Stefan; Funke, Harald; Mosig, Alexander

2015-03-02

Hemodynamic forces generated by the blood flow are of central importance for the function of endothelial cells (ECs), which form a biologically active cellular monolayer in blood vessels and serve as a selective barrier for macromolecular permeability. Mechanical stimulation of the endothelial monolayer induces morphological remodeling in its cytoskeleton. For in vitro studies on EC biology culture devices are desirable that simulate conditions of flow in blood vessels and allow flow-based adhesion/permeability assays under optimal perfusion conditions. With this aim we designed a biochip comprising a perfusable membrane that serves as cell culture platform multi-organ-tissue-flow (MOTiF biochip). This biochip allows an effective supply with nutrition medium, discharge of catabolic cell metabolites and defined application of shear stress to ECs under laminar flow conditions. To characterize EC layers cultured in the MOTiF biochip we investigated cell viability, expression of EC marker proteins and cell adhesion molecules of ECs dynamically cultured under low and high shear stress, and compared them with an endothelial culture in established two-dimensionally perfused flow chambers and under static conditions. We show that ECs cultured in the MOTiF biochip form a tight EC monolayer with increased cellular density, enhanced cell layer thickness, presumably as the result of a rapid and effective adaption to shear stress by remodeling of the cytoskeleton. Moreover, endothelial layers in the MOTiF biochip express higher amounts of EC marker proteins von-Willebrand-factor and PECAM-1. EC layers were highly responsive to stimulation with TNFα as detected at the level of ICAM-1, VCAM-1 and E-selectin expression and modulation of endothelial permeability in response to TNFα/IFNγ treatment under flow conditions. Compared to static and two-dimensionally perfused cell culture condition we consider MOTiF biochips as a valuable tool for studying EC biology in vitro under

Properties of Dental Pulp-derived Mesenchymal Stem Cells and the Effects of Culture Conditions.

PubMed

Kawashima, Nobuyuki; Noda, Sonoko; Yamamoto, Mioko; Okiji, Takashi

2017-09-01

Dental pulp mesenchymal stem cells (DPMSCs) highly express mesenchymal stem cell markers and possess the potential to differentiate into neural cells, osteoblasts, adipocytes, and chondrocytes. Thus, DPMSCs are considered suitable for tissue regeneration. The colony isolation method has commonly been used to collect relatively large amounts of heterogeneous DPMSCs. Homogenous DPMSCs can be isolated by fluorescence-activated cell sorting using antibodies against mesenchymal stem cell markers, although this method yields a limited number of cells. Both quality and quantity of DPMSCs are critical to regenerative therapy, and cell culture methods need to be improved. We thus investigated the properties of DPMSCs cultured with different methods. DPMSCs in a three-dimensional spheroid culture system, which is similar to the hanging drop culture for differentiation of embryonic stem cells, showed upregulation of odonto-/osteoblastic markers and mineralized nodule formation. This suggests that this three-dimensional spheroid culturing system for DPMSCs may be suitable for inducing hard tissues. We further examined the effect of cell culture density on the properties of DPMSCs because the properties of stem cells can be altered depending on the cell density. DPMSCs cultured under the confluent cell density condition showed slight downregulation of some mesenchymal stem cell markers compared with those under the sparse condition. The ability of DPMSCs to differentiate into hard tissue-forming cells was found to be enhanced in the confluent condition, suggesting that the confluent culture condition may not be suitable for maintaining the stemness of DPMSCs. When DPMSCs are to be used for hard tissue regeneration, dense followed by sparse cell culture conditions may be a better alternative strategy. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Response of an algal consortium to diesel under varying culture conditions.

PubMed

Chavan, Anal; Mukherji, Suparna

2010-03-01

A diesel-tolerant sessile freshwater algal consortium obtained from the vicinity of Powai Lake (Mumbai, India) was cultured in the laboratory. The presence of diesel in batch cultures enhanced the maximum specific growth rate of the algal consortium. With decrease in light-dark (L:D) cycle from 20:4 to 4:20 h, the chlorophyll-a levels decreased; however, the removal of diesel was found to be maximum at L:D of 18:6 h with 37.6% degradation over and above controls. In addition to growth in the form of green clumps, white floating biomass was found surrounding the diesel droplets on the surface. This culture predominated at the least L:D ratio of 4:20 h. Studies confirmed the ability of the floating organisms to grow heterotrophically in the dark utilizing diesel as carbon source and also in the presence of light in a medium devoid of organic carbon sources.

Microbiological characteristics of a sandy loam soil exposed to tebuconazole and lambda-cyhalothrin under laboratory conditions.

PubMed

Cycoń, M; Piotrowska-Seget, Z; Kaczyńska, A; Kozdrój, J

2006-11-01

Changes in microbiological properties of a sandy loam soil in response to the addition of different concentrations of fungicide tebuconazole and pyrethroid insecticide lambda-cyhalothrin were assessed under laboratory conditions. To ascertain these changes, the potentially active soil microbial biomass, concentrations of ammonium and nitrate ions, numbers of total culturable bacteria, fungi, nitrogen-fixing bacteria, nitrifying and denitrifying bacteria were determined. Substrate-induced respiration (SIR) increased with time in both control (ranged from 13.7 to 23.7 mg/O(2)/kg(-1)/dry soil/h(-1)) and pesticide treated soil portions. For both pesticides, SIR values ranged from 12-13 to 23-25 mg/O(2)/kg(-1)/dry soil/h(-1) on days 1 and 28, respectively. Also, concentrations of nitrate and ammonium ions, numbers of total culturable bacteria, denitrifying bacteria, nitrogen-fixing bacteria (for the insecticide) and fungi (for the insecticide) were either unaffected or even stimulated by the pesticide treatments. The adverse impacts of the pesticides were observed for nitrate concentrations (on days 1 or 7), numbers of nitrifying bacteria (on day 1), denitrifying bacteria (for the insecticide on days 1 and 14), nitrogen-fixing bacteria (for tebuconazole on day 1) as well as numbers of fungi in tebuconazole-treated soil (on days 1 and 14).

Culture Conditions for Mycelial Growth of Coriolus versicolor

PubMed Central

Kang, Min-Jin; Choi, Seong-Yong; Yoo, Young-Bok; Seok, Soon-Ja; Jung, Hee-Young

2010-01-01

Coriolus versicolor, is one of the most popular medicinal mushrooms due its various biologically active components. This study was conducted to obtain basic information regarding the mycelial culture conditions of C. versicolor. Based on the culture, and MCM media were suitable for the mycelial growth of the mushroom. The optimum carbon and nitrogen sources were dextrin and yeast extract, respectively, and the optimum C/N ratio was 10 to 2 when 2% glucose was used. Other minor components required for optimal growth included thiamine-HCl and biotin as vitamins, succinic acid, lactic acid and citric acid as organic acids, as well as MgSO4·7H2O as mineral salts. PMID:23956654

Culture Conditions for Mycelial Growth of Coriolus versicolor.

PubMed

Jo, Woo-Sik; Kang, Min-Jin; Choi, Seong-Yong; Yoo, Young-Bok; Seok, Soon-Ja; Jung, Hee-Young

2010-09-01

Coriolus versicolor, is one of the most popular medicinal mushrooms due its various biologically active components. This study was conducted to obtain basic information regarding the mycelial culture conditions of C. versicolor. Based on the culture, and MCM media were suitable for the mycelial growth of the mushroom. The optimum carbon and nitrogen sources were dextrin and yeast extract, respectively, and the optimum C/N ratio was 10 to 2 when 2% glucose was used. Other minor components required for optimal growth included thiamine-HCl and biotin as vitamins, succinic acid, lactic acid and citric acid as organic acids, as well as MgSO4·7H2O as mineral salts.

Cell death in Tetrahymena thermophila: new observations on culture conditions.

PubMed

Christensen, S T; Sørensen, H; Beyer, N H; Kristiansen, K; Rasmussen, L; Rasmussen, M I

2001-01-01

We previously suggested that the cell fate of the protozoan ciliate, Tetrahymena thermophila, effectively relates to a quorum-sensing mechanism where cell-released factors support cell survival and proliferation. The cells have to be present above a critical initial density in a chemically defined nutrient medium in order to release a sufficient level of these factors to allow a new colony to flourish. At a relatively high rate of metabolism and/or macromolecular synthesis and below this critical density, cells began to die abruptly within 30 min of inoculation, and this death took the form of an explosive disintegration lasting less than 50 milliseconds. The cells died at any location in the culture, and the frequency of cell death was always lower in well-filled vials than those with medium/air interface. Cell death was inhibited by the addition of Actinomycin D or through modifications of the culture conditions either by reducing the oxygen tension or by decreasing the temperature of the growth medium. In addition, plastic caps in well-filled vials release substances, which promote cell survival. The fate of low-density cultures is related to certain 'physical' conditions, in addition to the availability of oxygen within closed culture systems. Copyright 2001 Academic Press.

Persistence in soil of Miscanthus biochar in laboratory and field conditions

PubMed Central

Budai, Alice; O’Toole, Adam; Ma, Xingzhu; Rumpel, Cornelia; Abiven, Samuel

2017-01-01

Evaluating biochars for their persistence in soil under field conditions is an important step towards their implementation for carbon sequestration. Current evaluations might be biased because the vast majority of studies are short-term laboratory incubations of biochars produced in laboratory-scale pyrolyzers. Here our objective was to investigate the stability of a biochar produced with a medium-scale pyrolyzer, first through laboratory characterization and stability tests and then through field experiment. We also aimed at relating properties of this medium-scale biochar to that of a laboratory-made biochar with the same feedstock. Biochars were made of Miscanthus biomass for isotopic C-tracing purposes and produced at temperatures between 600 and 700°C. The aromaticity and degree of condensation of aromatic rings of the medium-scale biochar was high, as was its resistance to chemical oxidation. In a 90-day laboratory incubation, cumulative mineralization was 0.1% for the medium-scale biochar vs. 45% for the Miscanthus feedstock, pointing to the absence of labile C pool in the biochar. These stability results were very close to those obtained for biochar produced at laboratory-scale, suggesting that upscaling from laboratory to medium-scale pyrolyzers had little effect on biochar stability. In the field, the medium-scale biochar applied at up to 25 t C ha-1 decomposed at an estimated 0.8% per year. In conclusion, our biochar scored high on stability indices in the laboratory and displayed a mean residence time > 100 years in the field, which is the threshold for permanent removal in C sequestration projects. PMID:28873471

Transformation of Corporate Culture in Conditions of Transition to Knowledge Economics

ERIC Educational Resources Information Center

Korsakova, Tatiana V.; Chelnokova, Elena A.; Kaznacheeva, Svetlana N.; Bicheva, Irena B.; Lazutina, Antonina L.; Perova, Tatyana V.

2016-01-01

This article is devoted to the problem of corporate culture transformations which are conditioned by changes in social-economic situation. The modern paradigm of knowledge management is assumed to become the main value for forming a new vision of corporate culture. The starting point for transformations can be found in the actual corporate culture…

Procedure for Adaptive Laboratory Evolution of Microorganisms Using a Chemostat.

PubMed

Jeong, Haeyoung; Lee, Sang J; Kim, Pil

2016-09-20

Natural evolution involves genetic diversity such as environmental change and a selection between small populations. Adaptive laboratory evolution (ALE) refers to the experimental situation in which evolution is observed using living organisms under controlled conditions and stressors; organisms are thereby artificially forced to make evolutionary changes. Microorganisms are subject to a variety of stressors in the environment and are capable of regulating certain stress-inducible proteins to increase their chances of survival. Naturally occurring spontaneous mutations bring about changes in a microorganism's genome that affect its chances of survival. Long-term exposure to chemostat culture provokes an accumulation of spontaneous mutations and renders the most adaptable strain dominant. Compared to the colony transfer and serial transfer methods, chemostat culture entails the highest number of cell divisions and, therefore, the highest number of diverse populations. Although chemostat culture for ALE requires more complicated culture devices, it is less labor intensive once the operation begins. Comparative genomic and transcriptome analyses of the adapted strain provide evolutionary clues as to how the stressors contribute to mutations that overcome the stress. The goal of the current paper is to bring about accelerated evolution of microorganisms under controlled laboratory conditions.

An optimal serum-free defined condition for in vitro culture of kidney organoids.

PubMed

Nishikawa, Masaki; Kimura, Hiroshi; Yanagawa, Naomi; Hamon, Morgan; Hauser, Peter; Zhao, Lifu; Jo, Oak D; Yanagawa, Norimoto

2018-07-02

Kidney organoid is an emerging topic of importance for research in kidney development and regeneration. Conventional culture systems for kidney organoids reported thus far use culture media containing serum, which may compromise our understanding and the potential clinical applicability of the organoid system. In our present study, we tested two serum-free culture conditions and compared their suitability for the maintenance and growth of kidney organoids in culture. One of the serum-free culture conditions was the combination of keratinocytes serum free medium (KSFM) with knockout serum replacement (KSR) (KSFM + KSR), and the other was the combination of knockout DMEM/F12 (KD/F12) and KSR (KD/F12 + KSR). With cell aggregates derived from E12.5 mouse embryonic kidneys, we found that KD/F12 + KSR was superior to KSFM + KSR in promoting the growth of the aggregate with expansion of Six2 + nephron progenitor cells (NPC) and elaborated ureteric branching morphogenesis. With KD/F12 + KSR, we found that lower concentrations of KSR at 5-10% were superior to a higher concentration (20%) in promoting the growth of aggregates without affecting the expression levels of NPC marker genes. We also found that NPC in aggregates retained their differentiation potential to develop nephron tubules through mesenchyme-to-epithelial transition (MET), after being maintained in culture under these conditions for up to 7 days. In conclusion, we have identified a defined serum-free culture condition suitable for the maintenance and growth of kidney organoids that retain the differentiation potential to develop nephron structures. This defined serum-free culture condition may serve as a useful platform for further investigation of kidney organoids in vitro. Copyright © 2018 Elsevier Inc. All rights reserved.

Cholera toxin expression by El Tor Vibrio cholerae in shallow culture growth conditions.

PubMed

Cobaxin, Mayra; Martínez, Haydee; Ayala, Guadalupe; Holmgren, Jan; Sjöling, Asa; Sánchez, Joaquín

2014-01-01

Vibrio cholerae O1 classical, El Tor and O139 are the primary biotypes that cause epidemic cholera, and they also express cholera toxin (CT). Although classical V. cholerae produces CT in various settings, the El Tor and O139 strains require specific growth conditions for CT induction, such as the so-called AKI conditions, which consist of growth in static conditions followed by growth under aerobic shaking conditions. However, our group has demonstrated that CT production may also take place in shallow static cultures. How these type of cultures induce CT production has been unclear, but we now report that in shallow culture growth conditions, there is virtual depletion of dissolved oxygen after 2.5 h of growth. Concurrently, during the first three to 4 h, endogenous CO2 accumulates in the media and the pH decreases. These findings may explain CT expression at the molecular level because CT production relies on a regulatory cascade, in which the key regulator AphB may be activated by anaerobiosis and by low pH. AphB activation stimulates TcpP synthesis, which induces ToxT production, and ToxT directly stimulates ctxAB expression, which encodes CT. Importantly, ToxT activity is enhanced by bicarbonate. Therefore, we suggest that in shallow cultures, AphB is activated by initial decreases in oxygen and pH, and subsequently, ToxT is activated by intracellular bicarbonate that has been generated from endogenous CO2. This working model would explain CT production in shallow cultures and, possibly, also in other growth conditions. Copyright © 2013 Elsevier Ltd. All rights reserved.

Inter-laboratory comparison of radiometric culture for Mycobacterium avium subsp. paratuberculosis using raw milk from known infected herds and individual dairy cattle in Victoria.

PubMed

Ridge, S E; Andreata, S; Jones, K; Cantlon, K; Francis, B; Florisson, N; Gwozdz, J

2010-07-01

To compare the results of radiometric culture conducted in three Australian laboratories for Mycobacterium avium subsp. paratuberculosis (Mptb) using bulk vat and individual animal milk samples. Milk samples were collected from 15 cows exhibiting clinical signs of Johne's disease, and subsequently confirmed as infected with Mptb, and from the bulk milk vats on 91 farms running herds known to be infected with Mptb. Each milk sample was divided into three equivalent samples and one of each of the replicates was forwarded to the three participating laboratories. The identity and nature of the samples was protected from the study collaborators. The laboratories processed the samples and undertook radiometric culture for Mptb using their standard method. Results of testing were provided to the principal investigator for collation and analysis. In total, 2 (2.2%) of 91 vat-milk samples and 8 (53.3%) of 15 individual cows' milk samples returned positive radiometric milk culture results. Only one sample, from a clinical case of Johne's disease, was identified as positive by more than one laboratory. There were differences in the absolute frequency with which Mptb was identified in the milk samples by the collaborating laboratories. Mptb was cultured from a very small percentage of Australian raw bulk milk samples sourced from known infected herds. By contrast, Mptb was successfully cultured from half of the milk samples collected from clinically affected cows. There was no statistical difference between laboratories in the proportion of vat samples or individual animal milk samples in which Mptb was detected.

Laboratory evaluation and application of microwave absorption properties under simulated conditions for planetary atmospheres

NASA Technical Reports Server (NTRS)

Steffes, Paul G.

1991-01-01

Laboratory measurements of microwave and millimeter wave properties of the simulated atmosphere of the outer planets and their satellites has continued. One of the focuses is on the development of a radiative transfer model of the Jovian atmosphere at wavelengths from 1 mm to 10 cm. This modeling effort led to laboratory measurements of the millimeter wave opacity of hydrogen sulfide (H2S) under simulated Jovian conditions. Descriptions of the modeling effort, the Laboratory experiment, and the observations are presented. Correlative studies of measurements with Pioneer-Venus radio occultation measurements with longer wavelength emission measurements have provided new ways for characterizing temporal and spatial variations in the abundance of both gases H2SO4 and SO2, and for modeling their roles in the subcloud atmosphere. Laboratory measurements were conducted on 1.35 cm (and 13 cm) opacity of gaseous SO2 and absorptivity of gaseous SO2 at the 3.2 mm wavelength under simulated Venus conditions. Laboratory measurements were completed on millimeter wave dielectric properties of liquid H2SO4, in order to model the effects of the opacity of the clouds of Venus onto millimeter wave emission spectrum.

Isolation and Culture of Bovine Oviductal Epithelial Cells for Use in the Anatomy and Physiology Laboratory and Undergraduate Research

ERIC Educational Resources Information Center

Way, Amy L.

2006-01-01

This article presents methods for the isolation and culture of epithelial cells from the bovine oviduct for use in both research and the teaching laboratory and provides examples of ways that an oviductal cell culture can be incorporated into an undergraduate research program. Cow reproductive tracts are readily available from area butchers, and…

Experiments with osteoblasts cultured under hypergravity conditions

NASA Technical Reports Server (NTRS)

Kacena, Melissa A.; Todd, Paul; Gerstenfeld, Louis C.; Landis, William J.

2004-01-01

To understand further the role of gravity in osteoblast attachment, osteoblasts were subjected to hypergravity conditions in vitro. Scanning electron microscopy of all confluent coverslips from FPA units show that the number of attached osteoblasts was similar among gravitational levels and growth durations (90 cells/microscopic field). Specifically, confluent 1.0 G control cultures contained an average of 91 +/- 8 cells/field, 3.3 G samples had 88 +/- 8 cells/field, and 4.0 G cultures averaged 90 +/- 7 cells/field. The sparsely plated cultures assessed by immunohistochemistry also had similar numbers of cells at each time point (l.0 G was similar to 3.3 and 4.0 G), but cell number changed from one time point to the next as those cells proliferated. Immunohistochemistry of centrifuged samples showed an increase in number (up to 160% increase) and thickness (up to 49% increase) of actin fibers, a decrease in intensity of fibronectin fluorescence (18-23% decrease) and an increase in number of vinculin bulbs (202-374% increase in number of vinculin bulbs/area). While hypergravity exposure did not alter the number of attached osteoblasts, it did result in altered actin, fibronectin, and vinculin elements, changing some aspects of osteoblast- substrate adhesion.

Influence of culture conditions for clinically isolated non-albicans Candida biofilm formation.

PubMed

Tan, Yulong; Leonhard, Matthias; Ma, Su; Schneider-Stickler, Berit

2016-11-01

Non-albicans Candida species have been isolated in increasing numbers in patients. Moreover, they are adept at forming biofilms. This study analyzed biofilm formation of clinically isolated non-albicans Candida, including Candida tropicalis, Candida krusei and Candida parapsilosis under the influence of different growth media (RPMI 1640, YPD and BHI) and several culture variables (inoculum concentration, incubation period and feeding conditions). The results showed that culture conditions strongly influenced non-albicans Candida species biofilm formation. YPD and BHI resulted in larger amount of biofilm formation with higher metabolic activity of biofilms. Furthermore, the growth media seems to have varying effects on adhesion and biofilm development. Growth conditions may also influence biofilm formation, which was enhanced when starting the culture with a larger inoculum, longer incubation period and using a fed-batch system. Therefore, the potential influences of external environmental factors should be considered when studying the non-albicans Candida biofilms in vitro. Copyright © 2016 Elsevier B.V. All rights reserved.

Evaluation of foal production following intracytoplasmic sperm injection and blastocyst culture of oocytes from ovaries collected immediately before euthanasia or after death of mares under field conditions.

PubMed

Hinrichs, Katrin; Choi, Young-Ho; Norris, Jody D; Love, Linda B; Bedford-Guaus, Sylvia J; Hartman, David L; Velez, Isabel C

2012-10-15

To evaluate the efficiency of foal production following intracytoplasmic sperm injection (ICSI) and blastocyst culture of oocytes from mares that died or were euthanized under field conditions. Prospective case series. 16 mares (age, 3 to 19 years) that died or were euthanized for various causes. Ovaries were collected immediately before euthanasia (n = 10) or after death (6). Ovaries were transported to the laboratory for oocyte recovery (15 mares), or oocytes were recovered at a remote location and shipped to the laboratory (1). Oocytes underwent ICSI, and presumptive zygotes were cultured for 7 to 10 days. Blastocysts were shipped to embryo transfer facilities for transcervical transfer to recipient mares. Ovaries were processed 30 minutes to 12 hours (mean ± SD, 4.6 ± 3.3 hours) after mares' deaths. A mean of 14.1 ± 8.6 oocytes/mare were cultured, and 110 of 225 (49%) matured. Twenty-one blastocysts developed after ICSI and were transferred to recipient mares. Thirteen pregnancies were established; 10 healthy foals were produced from 6 donor mares. The number of blastocysts produced per mare and number of live foals produced per mare were significantly correlated with the number of oocytes recovered. Foals were produced from mares after death or euthanasia under field conditions. Proportions of foals born overall (10 foals/16 mares) and mares from which ≥ 1 foal was produced (6/16) were greater than those reported following recovery and oviductal transfer of oocytes to inseminated recipients after death of donor mares under field conditions.

Community dynamics and metabolite target analysis of spontaneous, backslopped barley sourdough fermentations under laboratory and bakery conditions.

PubMed

Harth, Henning; Van Kerrebroeck, Simon; De Vuyst, Luc

2016-07-02

Barley flour is not commonly used for baking because of its negative effects on bread dough rheology and loaf volume. However, barley sourdoughs are promising ingredients to produce improved barley-based breads. Spontaneous barley sourdough fermentations were performed through backslopping (every 24h, 10days) under laboratory (fermentors, controlled temperature of 30°C, high dough yield of 400) and bakery conditions (open vessels, ambient temperature of 17-22°C, low dough yield of 200), making use of the same batch of flour. They differed in pH evolution, microbial community dynamics, and lactic acid bacteria (LAB) species composition. After ten backsloppings, the barley sourdoughs were characterized by the presence of the LAB species Lactobacillus fermentum, Lactobacillus plantarum, and Lactobacillus brevis in the case of the laboratory productions (fast pH decrease, pH<4.0 after two backslopping steps), and of Leuconostoc citreum, Leuconostoc mesenteroides, Weissella confusa and Weissella cibaria in the case of the bakery productions (slow pH decrease, pH4.0 after eight backslopping steps). In both sourdough productions, Saccharomyces cerevisiae was the sole yeast species. Breads made with wheat flour supplemented with 20% (on flour basis) barley sourdough displayed a firmer texture, a smaller volume, and an acceptable flavour compared with all wheat-based reference breads. Hence, representative strains of the LAB species mentioned above, adapted to the environmental conditions they will be confronted with, may be selected as starter cultures for the production of stable barley sourdoughs and flavourful breads. Copyright © 2016. Published by Elsevier B.V.

[The opportunity to use combined stem cells transplantation for haemopoesis activation in the old and mature laboratory animals under the conditions of ionizing radiation].

PubMed

Grebnev, D U

2014-01-01

The objective of this work was to study the influence of combined transplantation of stem cells (multypotent mesenchimal stromal and hem poetic stem cells) on the haemopoesis of old and mature laboratory animals under the condition of ionizing radiation. The experiments were conducted on 48 white male mice with the body weight of 30 g, age of 3-4 months, and 48 male mice of 3 years of age and body mass of 50 g. The experiments for obtaining the MMSC and HSC cultures were conducted on 16 laboratory animals: female mice of 3-4 months of age and body mass of 30 g., 18 days gestation period. The control group was formed by the animals not under the ionizing radiation. The experimental group animals got the dose of 4 Gr. These animals also got MMSC and HSC mixture intravenously in the doses of 6 mln. c/kg. and 330 thousand cell/kg prospectively. The control group animals got the 0.9% NaCl - 0.2 ml. intravenously. The infusions were made 1 hour after radiation once. As the result of the experiment it was shown that under physiological conditions combined transplantation brings the erithropoesis activation, under the ionizing radiation conditions it brings the erythroid and granulocytopoesis activation. More over the combined MMSC and HSC transplantation gives cytoprotective action on the myeloid tissue due to decrease of cyto genically changed cells in the mature animals under the condition of ionizing radiation, but in the old animals this effect can be seen even under physiological condition. Conclusions: Combined transplantation of MMSC and GSC can be used in the mature and old laboratory animals under the conditions of ionising radiation for the haemopoesis activation.

GUIDANCE UNITS FOR THE LEARNING LABORATORY TO TEACH BASIC SKILLS IN A CULTURALLY DEPRIVED AREA.

ERIC Educational Resources Information Center

Dade County Public Schools, Miami, FL.

THE PURPOSE OF THIS HANDBOOK IS TO PROVIDE GUIDANCE UNITS FOR THE LEARNING LABORATORY. THE 10 UNITS ARE STRUCTURED TO TEACH BASIC SKILLS TO CULTURALLY DISADVANTAGED STUDENTS. THE FOLLOWING AREAS ARE SUBJECTS FOR INSTRUCTIONAL UNITS OF STUDY--(1) EXPLORING THE SELF-CONCEPT, (2) ATTITUDES, (3) HOW TO STUDY, (4) HOW TO PASS EXAMINATIONS, (5) GROUP…

Light field and water clarity simulation of natural environments in laboratory conditions

NASA Astrophysics Data System (ADS)

Pe'eri, Shachak; Shwaery, Glenn

2012-06-01

Simulation of natural oceanic conditions in a laboratory setting is a challenging task, especially when that environment can be miles away. We present an attempt to replicate the solar radiation expected at different latitudes with varying water clarity conditions up to 30 m in depth using a 2.5 m deep engineering tank at the University of New Hampshire. The goals of the study were: 1) to configure an underwater light source that produced an irradiance spectrum similar to natural daylight with the sun at zenith and at 60° under clear atmospheric conditions, and 2) to monitor water clarity as a function of depth. Irradiance was measured using a spectra-radiometer with a cosine receiver to analyze the output spectrum of submersed lamps as a function of distance. In addition, an underwater reflection method was developed to measure the diffuse attenuation coefficient in real time. Two water clarity types were characterized, clear waters representing deep, open-ocean conditions, and murky waters representing littoral environments. Results showed good correlation between the irradiance measured at 400 nm to 600 nm and the natural daylight spectrum at 3 m from the light source. This can be considered the water surface conditions reference. Using these methodologies in a controlled laboratory setting, we are able to replicate illumination and water conditions to study the physical, chemical and biological processes on natural and man-made objects and/or systems in simulated, varied geographic locations and environments.

Monochloramine Cometabolism by Mixed-Culture Nitrifiers under Drinking Water Conditions

EPA Science Inventory

The current research investigated monochloramine cometabolism by nitrifying mixed cultures grown under drinking water relevant conditions and harvested from sand-packed reactors before conducting suspended growth batch kinetic experiments. Three batch reactors were used in each ...

42 CFR 493.1409 - Condition: Laboratories performing moderate complexity testing; technical consultant.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Laboratories performing moderate complexity testing; technical consultant. 493.1409 Section 493.1409 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION...

42 CFR 493.1421 - Condition: Laboratories performing moderate complexity testing; testing personnel.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Laboratories performing moderate complexity testing; testing personnel. 493.1421 Section 493.1421 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION...

42 CFR 493.1409 - Condition: Laboratories performing moderate complexity testing; technical consultant.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing moderate complexity testing; technical consultant. 493.1409 Section 493.1409 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION...

42 CFR 493.1415 - Condition: Laboratories performing moderate complexity testing; clinical consultant.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing moderate complexity testing; clinical consultant. 493.1415 Section 493.1415 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION...

42 CFR 493.1421 - Condition: Laboratories performing moderate complexity testing; testing personnel.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 5 2011-10-01 2011-10-01 false Condition: Laboratories performing moderate complexity testing; testing personnel. 493.1421 Section 493.1421 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION...

Language Personality in the Conditions of Cross-Cultural Communication: Case-Study Experience

ERIC Educational Resources Information Center

Davidovitch, Nitza; Khyhniak, Kateryna

2018-01-01

The article is devoted to the problem of identification of a language personality's traits under conditions of cross-cultural communication. It is shown that effective cross-cultural communication is revised under globalization and increasingly intensive social interactions. The results of the authors' research prove that it is possible to develop…

Volatile organic compounds and good laboratory practices in the in vitro fertilization laboratory: the important parameters for successful outcome in extended culture.

PubMed

Agarwal, Nupur; Chattopadhyay, Ratna; Ghosh, Sanghamitra; Bhoumik, Arpita; Goswami, S K; Chakravarty, Baidyanath

2017-08-01

This study aims to describe the role of implementing good laboratory practices to improve in vitro fertilization (IVF) outcomes which are of great interest for practitioners dealing with infertility. Certain modifications were introduced in May 2015 in our IVF laboratory like high-efficiency particulate air CODA system, steel furniture instead of wooden, use of new disinfectants like oosafe, and restriction of personnel entry along with avoidance of cosmetics like perfume to improve pregnancy rates. Volatile organic compound (VOC) meter reading was monitored at two time points and five different places in the laboratory to compare the embryonic development parameters before (group A: July 2014-April 2015) and after (group B: July 2015-April 2016) remodeling. The IVF outcomes from 1036 cycles were associated in this study. Reduction in VOC meter readings, enhanced air quality, improvement in blastocyst formation rate, implantation, and clinical pregnancy rate were observed in the laboratory after implementation of new facilities. Results illustrated that the attention must be focused on potential hazards which expose laboratories to elevated VOC levels. Blastocyst formation rate increased around 18%. Implantation rate, clinical pregnancy rate, and live birth rate increased by around 11, 10, and 8%, respectively. In conclusion, with proper engineering and material selection, we have been able to reduce chemical contamination and adverse effects on culture with optimized IVF results. None.

The Tissue Culture Laboratory of Dr. George Otto Gey 60 yrs ago as recalled by a former student.

PubMed

Ambrose, Charles T

2017-05-01

George Otto Gey was a pioneer in tissue culture, having introduced the roller drum, the HeLa cell line, and the use of human fetal cord serum and beef embryo extract. During his career (1920s-1960s), the field of tissue culture was in its infancy and not yet dependent upon commercial biological supply houses. While the early techniques of cell culture have been greatly improved upon, of historical interest may be personal observations of the Gey Tissue Culture Laboratory, Johns Hopkins Medical School, as recalled by a medical student working there in the 1950s. Dr. Gey served as a founding member and executive of the Tissue Culture Commission (TCC) and became the first president of the Tissue Culture Association (TCA).

Use of cultured human epidermal keratinocytes for allografting burns and conditions for temporary banking of the cultured allografts.

PubMed

Bolívar-Flores, J; Poumian, E; Marsch-Moreno, M; Montes de Oca, G; Kuri-Harcuch, W

1990-02-01

Five children who suffered burns clinically regarded as full skin thickness loss were grafted with cultured allogeneic skin from newborn prepuce. The wounds had remained open and infected without healing for about 20 days before the patients were received in the burn unit. To avoid losing surviving deep epidermal cells the wounds were débrided but not deeply excised and, a few days before allografting, they were washed with isodine solution and sterile water, and treated with silvadene cream application. All children received 76 cultured allografts of about 60 cm2 each. After allografting, the wounds were epithelized in 7-10 days and the allogeneic grafted skin began desquamation suggesting that the allograft did not 'take' permanently but was replaced by the newly formed skin. On the other hand, since allografting is an adequate therapy to provide early temporary coverage in extensively burned patients, we developed conditions for banking cultured skin to make it available for immediate use. The conditions described allow banking of the cultured grafts for 15-20 days with retention of clonal growth ability similar to that of unstored epithelia. The results show that cultured epidermal cells obtained from human newborn foreskin, when used as allografts for coverage of full skin or deep partial skin thickness burns, allow rapid epithelization of the burn wounds.

Mycotoxin production of selected Fusarium species at different culture conditions.

PubMed

Kokkonen, Meri; Ojala, Laura; Parikka, Päivi; Jestoi, Marika

2010-09-30

The toxin producing capacity of seven Fusarium species (F. langsethiae, F. sporotrichioides, F. poae, F. avenaceum, F. tricinctum, F. graminearum and F. culmorum) and the effect of culture conditions on the toxin production were studied. The strains were isolated from Finnish grains and cultivated on a grain mixture at three different water activity/temperature combinations (i.e. 0.994/15 degrees C; 0.994/25 degrees C; 0.960/25 degrees C). The mycotoxins produced were analyzed with a multi-toxin method based on liquid chromatography-tandem mass spectrometry enabling the simultaneous determination of 18 different Fusarium toxins. The general toxin profiles revealed F. langsethiae and F. sporotrichioides as producers of diacetoxyscirpenol, neosolaniol, HT-2 and T-2-toxins. F. sporotrichioides produced additionally beauvericin. In the F. poae cultures, only beauvericin was detected. F. avenaceum and F. tricinctum were capable of producing enniatins, moniliformin and antibiotic Y, and F. graminearum and F. culmorum produced zearalenone, deoxynivalenol and 3-acetyl deoxynivalenol. Differences existed in the quantitative toxin production between the individual strains representing the same species. Additionally, the culture conditions affected the range and amounts of toxins produced. In general, a(w) 0.994 and temperature of 15 degrees C favoured the type-A trichothecene production of F. langsethiae and F. sporotrichioides. The beauvericin production of F. sporotrichioides occurred more favourably at a(w) 0.960 and 25 degrees C. F. poae produced the highest concentrations of beauvericin under two different conditions, namely at a(w) 0.994/15 degrees C and a(w) 0.960/25 degrees C. None of the combinations particularly favoured toxin production of F. avenaceum, with all three toxins being produced extensively at all culture conditions. F. tricinctum produced enniatins most efficiently at a(w) 0.994/25 degrees C. The moniliformin production of both these two species

Cultured astrocytes do not release adenosine during hypoxic conditions

PubMed Central

Fujita, Takumi; Williams, Erika K; Jensen, Tina K; Smith, Nathan A; Takano, Takahiro; Tieu, Kim; Nedergaard, Maiken

2012-01-01

Recent reports based on a chemiluminescent enzymatic assay for detection of adenosine conclude that cultured astrocytes release adenosine during mildly hypoxic conditions. If so, astrocytes may suppress neural activity in early stages of hypoxia. The aim of this study was to reevaluate the observation using high-performance liquid chromatography (HPLC). The HPLC analysis showed that exposure to 20 or 120 minutes of mild hypoxia failed to increase release of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), and adenosine from cultured astrocytes. Similar results were obtained using a chemiluminescent enzymatic assay. Moreover, since the chemiluminescent enzymatic assay relies on hydrogen peroxide generation, release of free-radical scavengers from hypoxic cells can interfere with the assay. Accordingly, adenosine added to samples collected from hypoxic cultures could not be detected using the chemiluminescent enzymatic assay. Furthermore, addition of free-radical scavengers sharply reduced the sensitivity of adenosine detection. Conversely, use of a single-step assay inflated measured values due to the inability of the assay to distinguish adenosine and its metabolite inosine. These results show that cultured astrocytes do not release adenosine during mild hypoxia, an observation consistent with their high resistance to hypoxia. PMID:21989480

Laboratory diagnosis of Chlamydia pneumoniae infections

PubMed Central

Peeling, Rosanna W

1995-01-01

Chlamydia pneumoniae is an important cause of respiratory illness. There is a need for accurate and rapid laboratory diagnostic methods that will lead to improved patient care, appropriate use of antimicrobial therapy and a better understanding of the epidemiology of this emerging pathogen. Culture is highly specific but is technically demanding, expensive, has a long turnaround time and its sensitivity is highly dependent on transport conditions. Antigen detection tests such as enzyme immunoassay and direct fluorescent antibody assay, and molecular detection methods such as the polymerase chain reaction assay, may provide a rapid diagnosis without the requirement for stringent transport conditions. The results of these tests should be interpreted with caution until more thorough evaluation is available. Serology remains the method of choice. The limitations of different serological methods for the laboratory diagnosis of C pneumoniae are discussed. PMID:22514397

Adherent culture conditions enrich the side population obtained from the cochlear modiolus-derived stem/progenitor cells.

PubMed

Chao, Ting-Ting; Wang, Chih-Hung; Chen, Hsin-Chien; Shih, Cheng-Ping; Sytwu, Huey-Kang; Huang, Kun-Lun; Chen, Shao-Yuan

2013-05-01

Previously, our group reported that sphere-forming cells derived from the organ of Corti represent the stem/progenitor cells (SPCs) of the cochlea due to their properties of self-renewal and multipotency. However, long-term propagation of sphere-forming cells under suspension culture conditions may fail to maintain the characteristic stemness of these cells. Therefore, this study investigated whether an adherent culture system would be beneficial in terms of preserving more stem-like cells for long-term manipulations in vitro. Isolated modiolus-derived SPCs were placed on poly-d-lysine-coated petri dishes to form the so-called "adherent" culture system. Modiolus SPCs cultured under adherent conditions exhibited a significantly increased percentage of cells with the side population (SP) phenotype (18.6%) compared with cells cultured under conventional suspension culture conditions (0.8%). Even after repeated passages, modiolus SPCs cultured under adherent culture conditions preserved more SP phenotype cells. In comparison with the non-SP phenotype cells, the sorted SP cells exhibited more stem-like but less differentiated properties, with an upregulated expression of the ATP-binding cassette subfamily G member 2 (ABCG2), Nestin, Sox2, and Nanog proteins. Furthermore, Retinoic acid (RA) treatment confirmed the expression of the multipotent differentiation markers in the SP cells, including TUJ1, pancytokeratin, glial fibrillary acidic protein (GFAP), and p27(Kip1). Employment of an adherent culture system, instead of a suspension culture system, resulted in the enrichment of the SP cells from SPCs while retaining their stemness and multipotency. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Cultural Resource Protection Plan for the Remote-Handled Low-Level Waste Disposal Facility at the Idaho National Laboratory

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

Pace, Brenda Ringe; Gilbert, Hollie Kae

2015-05-01

This plan addresses cultural resource protection procedures to be implemented during construction of the Remote Handled Low Level Waste project at the Idaho National Laboratory. The plan proposes pre-construction review of proposed ground disturbing activities to confirm avoidance of cultural resources. Depending on the final project footprint, cultural resource protection strategies might also include additional survey, protective fencing, cultural resource mapping and relocation of surface artifacts, collection of surface artifacts for permanent curation, confirmation of undisturbed historic canal segments outside the area of potential effects for construction, and/or archaeological test excavations to assess potential subsurface cultural deposits at known culturalmore » resource locations. Additionally, all initial ground disturbing activities will be monitored for subsurface cultural resource finds, cultural resource sensitivity training will be conducted for all construction field personnel, and a stop work procedure will be implemented to guide assessment and protection of any unanticipated discoveries after initial monitoring of ground disturbance.« less

Laboratory evaluation and application of microwave absorption properties under simulated conditions for planetary atmospheres

NASA Technical Reports Server (NTRS)

Steffes, Paul G.

1988-01-01

In the first half of this grant year, laboratory measurements were conducted on the millimeter-wave properties of atmospheric gases under simulated conditions for the outer planet. Significant improvements in the current system have made it possible to accurately characterize the opacity from gaseous NH3 at longer millimeter wavelengths (7 to 10 mm) under simulated Jovian conditions. In the second half of the grant year, it is hoped to extend such measurements to even shorter millimeter-wavelengths. Further analysis and application of the laboratory results to microwave and millimeter-wave absorption data for the outer planets, such as results from Voyager Radio Occultation experiments and earth-based radio astronomical observations will be continued. The analysis of available multispectral microwave opacity data from Venus, including data from the most recent radio astronomical ovservations in the 1.3 to 3.6 cm wavelength range and newly obtained Pioneer-Venus Radio Occulatation measurements at 13 cm, using the laboratory measurements as an interpretative tool will be pursued.

Human spermatogonial stem cells display limited proliferation in vitro under mouse spermatogonial stem cell culture conditions.

PubMed

Medrano, Jose V; Rombaut, Charlotte; Simon, Carlos; Pellicer, Antonio; Goossens, Ellen

2016-11-01

To study the ability of human spermatogonial stem cells (hSSCs) to proliferate in vitro under mouse spermatogonial stem cell (mSSC) culture conditions. Experimental basic science study. Reproductive biology laboratory. Cryopreserved testicular tissue with normal spermatogenesis obtained from three donors subjected to orchiectomy due to a prostate cancer treatment. Testicular cells used to create in vitro cell cultures corresponding to the following groups: [1] unsorted human testicular cells, [2] differentially plated human testicular cells, and [3] cells enriched with major histocompatibility complex class 1 (HLA - )/epithelial cell surface antigen (EPCAM + ) in coculture with inactivated testicular feeders from the same patient. Analyses and characterization including immunocytochemistry and quantitative reverse-transcription polymerase chain reaction for somatic and germ cell markers, testosterone and inhibin B quantification, and TUNEL assay. Putative hSSCs appeared in singlets, doublets, or small groups of up to four cells in vitro only when testicular cells were cultured in StemPro-34 medium supplemented with glial cell line-derived neurotrophic factor (GDNF), leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF). Fluorescence-activated cell sorting with HLA - /EPCAM + resulted in an enrichment of 27% VASA + /UTF1 + hSSCs, compared to 13% in unsorted controls. Coculture of sorted cells with inactivated testicular feeders gave rise to an average density of 112 hSSCs/cm 2 after 2 weeks in vitro compared with unsorted cells (61 hSSCs/cm 2 ) and differentially plated cells (49 hSSCS/cm 2 ). However, putative hSSCs rarely stained positive for the proliferation marker Ki67, and their presence was reduced to the point of almost disappearing after 4 weeks in vitro. We found that hSSCs show limited proliferation in vitro under mSSC culture conditions. Coculture of HLA - /EPCAM + sorted cells with testicular

Culture Conditions Affect Expression of DUX4 in FSHD Myoblasts.

PubMed

Pandey, Sachchida Nand; Khawaja, Hunain; Chen, Yi-Wen

2015-05-08

Facioscapulohumeral muscular dystrophy (FSHD) is believed to be caused by aberrant expression of double homeobox 4 (DUX4) due to epigenetic changes of the D4Z4 region at chromosome 4q35. Detecting DUX4 is challenging due to its stochastic expression pattern and low transcription level. In this study, we examined different cDNA synthesis strategies and the sensitivity for DUX4 detection. In addition, we investigated the effects of dexamethasone and knockout serum replacement (KOSR) on DUX4 expression in culture. Our data showed that DUX4 was consistently detected in cDNA samples synthesized using Superscript III. The sensitivity of DUX4 detection was higher in the samples synthesized using oligo(dT) primers compared to random hexamers. Adding dexamethasone to the culture media significantly suppressed DUX4 expression in immortalized (1.3 fold, p < 0.01) and primary (4.7 fold, p < 0.01) FSHD myoblasts, respectively. Culture medium with KOSR increased DUX4 expression and the response is concentration dependent. The findings suggest that detection strategies and culture conditions should be carefully considered when studying DUX4 in cultured cells.

Developing a lean culture in the laboratory.

PubMed

Napoles, Leyda; Quintana, Maria

2006-07-25

The Director of Pathology at Jackson Memorial Hospital was interested in improving the operational efficiencies of the department in order to enhance the department's level of service in conjunction with the expansion of the overall health system. The decision was made to implement proven Lean practices in the laboratory under the direction of a major consulting firm. This article details the scope of the initial project as well as the operating principles of Lean manufacturing practices as applied to the clinical laboratory. The goals of the project were to improve turnaround times of laboratory results, reduce inventory and supply costs, improve staff productivity, maximize workflow, and eliminate waste. Extensive data gathering and analysis guided the work process by highlighting the areas of highest opportunity. This systematic approach resulted in recommendations for the workflow and physical layout of the laboratory. It also included the introduction of "standard workflow" and "visual controls" as critical items that streamlined operational efficiencies. The authors provide actual photographs and schematics of the reorganization and improvements to the physical layout of the laboratory. In conclusion, this project resulted in decreased turnaround times and increased productivity, as well as significant savings in the overall laboratory operations.

Proteomic analysis of extracellular proteins from Aspergillus oryzae grown under submerged and solid-state culture conditions.

PubMed

Oda, Ken; Kakizono, Dararat; Yamada, Osamu; Iefuji, Haruyuki; Akita, Osamu; Iwashita, Kazuhiro

2006-05-01

Filamentous fungi are widely used for the production of homologous and heterologous proteins. Recently, there has been increasing interest in Aspergillus oryzae because of its ability to produce heterologous proteins in solid-state culture. To provide an overview of protein secretion by A. oryzae in solid-state culture, we carried out a comparative proteome analysis of extracellular proteins in solid-state and submerged (liquid) cultures. Extracellular proteins prepared from both cultures sequentially from 0 to 40 h were subjected to two-dimensional electrophoresis, and protein spots at 40 h were identified by peptide mass fingerprinting using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. We also attempted to identify cell wall-bound proteins of the submerged culture. We analyzed 85 spots from the solid-state culture and 110 spots from the submerged culture. We identified a total of 29 proteins, which were classified into 4 groups. Group 1 consisted of extracellular proteins specifically produced in the solid-state growth condition, such as glucoamylase B and alanyl dipeptidyl peptidase. Group 2 consisted of extracellular proteins specifically produced in the submerged condition, such as glucoamylase A (GlaA) and xylanase G2 (XynG2). Group 3 consisted of proteins produced in both conditions, such as xylanase G1. Group 4 consisted of proteins that were secreted to the medium in the solid-state growth condition but trapped in the cell wall in the submerged condition, such as alpha-amylase (TAA) and beta-glucosidase (Bgl). A Northern analysis of seven genes from the four groups suggested that the secretion of TAA and Bgl was regulated by trapping these proteins in the cell wall in submerged culture and that secretion of GlaA and XynG2 was regulated at the posttranscriptional level in the solid-state culture.

Proteomic Analysis of Extracellular Proteins from Aspergillus oryzae Grown under Submerged and Solid-State Culture Conditions

PubMed Central

Oda, Ken; Kakizono, Dararat; Yamada, Osamu; Iefuji, Haruyuki; Akita, Osamu; Iwashita, Kazuhiro

2006-01-01

Filamentous fungi are widely used for the production of homologous and heterologous proteins. Recently, there has been increasing interest in Aspergillus oryzae because of its ability to produce heterologous proteins in solid-state culture. To provide an overview of protein secretion by A. oryzae in solid-state culture, we carried out a comparative proteome analysis of extracellular proteins in solid-state and submerged (liquid) cultures. Extracellular proteins prepared from both cultures sequentially from 0 to 40 h were subjected to two-dimensional electrophoresis, and protein spots at 40 h were identified by peptide mass fingerprinting using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. We also attempted to identify cell wall-bound proteins of the submerged culture. We analyzed 85 spots from the solid-state culture and 110 spots from the submerged culture. We identified a total of 29 proteins, which were classified into 4 groups. Group 1 consisted of extracellular proteins specifically produced in the solid-state growth condition, such as glucoamylase B and alanyl dipeptidyl peptidase. Group 2 consisted of extracellular proteins specifically produced in the submerged condition, such as glucoamylase A (GlaA) and xylanase G2 (XynG2). Group 3 consisted of proteins produced in both conditions, such as xylanase G1. Group 4 consisted of proteins that were secreted to the medium in the solid-state growth condition but trapped in the cell wall in the submerged condition, such as α-amylase (TAA) and β-glucosidase (Bgl). A Northern analysis of seven genes from the four groups suggested that the secretion of TAA and Bgl was regulated by trapping these proteins in the cell wall in submerged culture and that secretion of GlaA and XynG2 was regulated at the posttranscriptional level in the solid-state culture. PMID:16672490

Laboratory evaluation and application of microwave absorption properties under simulated conditions for planetary atmospheres

NASA Technical Reports Server (NTRS)

Steffes, Paul G.

1987-01-01

Radio absorptivity data for planetary atmospheres obtained from spacecraft radio occultation experiments and Earth-based radio astronomical observations can be used to infer abundances of microwave absorbing atmospheric constituents in those atmospheres, as long as reliable information regarding the microwave absorping properties of potential constituents is available. The use of theoretically derived microwave absorption properties for such atmospheric constituents, or laboratory measurements of such properties under environmental conditions which are significantly different than those of the planetary atmosphere being studied, often leads to significant misinterpretation of available opacity data. Laboratory measurement of the microwave properties of atmospheric gases under simulated conditions for the outer planets were conducted. Results of these measurements are discussed.

Evolution of organic molecules under Mars-like UV radiation conditions in space and laboratory

NASA Astrophysics Data System (ADS)

Rouquette, L.; Stalport, F.; Cottin, H.; Coll, P.; Szopa, C.; Saiagh, K.; Poch, O.; Khalaf, D.; Chaput, D.; Grira, K.; Dequaire, T.

2017-09-01

The detection and identification of organic molecules at Mars are of prime importance, as some of these molecules are life precursors and components. While in situ planetary missions are searching for them, it is essential to understand how organic molecules evolve and are preserved at the surface of Mars. Indeed the harsh conditions of the environment of Mars such as ultraviolet (UV) radiation or oxidative processes could explain the low abundance and diversity of organic molecules detected by now [1]. In order to get a better understanding of the evolution of organic matter at the surface of Mars, we exposed organic molecules under a Mars-like UV radiation environment. Similar organic samples were exposed to the Sun radiation, outside the International Space Station (ISS), and under a UV lamp (martian pressure and temperature conditions) in the laboratory. In both experiments, organic molecules tend to photodegrade under Mars-like UV radiation. Minerals, depending on their nature, can protect or accelerate the degradation of organic molecules. For some molecules, new products, possibly photoresistant, seem to be produced. Finally, experimenting in space allow us to get close to in situ conditions and to validate our laboratory experiment while the laboratory experiment is essential to study the evolution of a large amount and diversity of organic molecules.

Laboratory versus outdoor cycling conditions: differences in pedaling biomechanics.

PubMed

Bertucci, William; Grappe, Frederic; Groslambert, Alain

2007-05-01

The aim of our study was to compare crank torque profile and perceived exertion between the Monark ergometer (818 E) and two outdoor cycling conditions: level ground and uphill road cycling. Seven male cyclists performed seven tests in seated position at different pedaling cadences: (a) in the laboratory at 60, 80, and 100 rpm; (b) on level terrain at 80 and 100 rpm; and (c) on uphill terrain (9.25% grade) at 60 and 80 rpm. The cyclists exercised for 1 min at their maximal aerobic power. The Monark ergometer and the bicycle were equipped with the SRM Training System (Schoberer, Germany) for the measurement of power output (W), torque (Nxm), pedaling cadence (rpm), and cycling velocity (kmxh-1). The most important findings of this study indicate that at maximal aerobic power the crank torque profiles in the Monark ergometer (818 E) were significantly different (especially on dead points of the crank cycle) and generate a higher perceived exertion compared with road cycling conditions.

Statistical optimization of culture conditions for bacterial cellulose production using Box-Behnken design.

PubMed

Bae, Sangok; Shoda, Makoto

2005-04-05

Culture conditions in a jar fermentor for bacterial cellulose (BC) production from A. xylinum BPR2001 were optimized by statistical analysis using Box-Behnken design. Response surface methodology was used to predict the levels of the factors, fructose (X1), corn steep liquor (CSL) (X2), dissolved oxygen (DO) (X3), and agar concentration (X4). Total 27 experimental runs by combination of each factor were carried out in a 10-L jar fermentor, and a three-dimensional response surface was generated to determine the effect of the factors and to find out the optimum concentration of each factor for maximum BC production and BC yield. The fructose and agar concentration highly influenced the BC production and BC yield. However, the optimum conditions according to changes in CSL and DO concentrations were predicted at almost central values of tested ranges. The predicted results showed that BC production was 14.3 g/L under the condition of 4.99% fructose, 2.85% CSL, 28.33% DO, and 0.38% agar concentration. On the other hand, BC yield was predicted in 0.34 g/g under the condition of 3.63% fructose, 2.90% CSL, 31.14% DO, and 0.42% agar concentration. Under optimized culture conditions, improvement of BC production and BC yield were experimentally confirmed, which increased 76% and 57%, respectively, compared to BC production and BC yield before optimizing the culture conditions. Copyright (c) 2005 Wiley Periodicals, Inc.

Full-life chronic toxicity of sodium salts to the mayfly Neocloeon triangulifer in tests with laboratory cultured food.

PubMed

Soucek, David J; Dickinson, Amy

2015-09-01

Although insects occur in nearly all freshwater ecosystems, few sensitive insect models exist for use in determining the toxicity of contaminants. The objectives of the present study were to adapt previously developed culturing and toxicity testing methods for the mayfly Neocloeon triangulifer (Ephemeroptera: Baetidae), and to further develop a method for chronic toxicity tests spanning organism ages of less than 24 h post hatch to adult emergence, using a laboratory cultured diatom diet. The authors conducted 96-h fed acute tests and full-life chronic toxicity tests with sodium chloride, sodium nitrate, and sodium sulfate. The authors generated 96-h median lethal concentrations (LC50s) of 1062 mg Cl/L (mean of 3 tests), 179 mg N-NO3 /L, and 1227 mg SO4 /L. Acute to chronic ratios ranged from 2.1 to 6.4 for chloride, 2.5 to 5.1 for nitrate, and 2.3 to 8.5 for sulfate. The endpoints related to survival and development time were consistently the most sensitive in the tests. The chronic values generated for chloride were in the same range as those generated by others using natural foods. Furthermore, our weight-versus-fecundity plots were similar to those previously published using the food culturing method on which the present authors' method was based, indicating good potential for standardization. The authors believe that the continued use of this sensitive mayfly species in laboratory studies will help to close the gap in understanding between standard laboratory toxicity test results and field-based observations of community impairment. © 2015 SETAC.

Bacterial Production of Poly(3-hydroxybutyrate): An Undergraduate Student Laboratory Experiment

ERIC Educational Resources Information Center

Burns, Kristi L.; Oldham, Charlie D.; May, Sheldon W.

2009-01-01

As part of a multidisciplinary course that is cross-listed between five departments, we developed an undergraduate student laboratory experiment for culturing, isolating, and purifying the biopolymer, poly(3-hydroxybutyrate), PHB. This biopolyester accumulates in the cytoplasm of bacterial cells under specific growth conditions, and it has…

Molecular biological and immunohistological characterization of canine dermal papilla cells and the evaluation of culture conditions.

PubMed

Kobayashi, Tetsuro; Fujisawa, Akiko; Amagai, Masayuki; Iwasaki, Toshiroh; Ohyama, Manabu

2011-10-01

The dermal papilla (DP) plays pivotal roles in hair follicle morphogenesis and cycling. However, our understanding of the biology of the canine DP is extremely limited. The aim of this study was to elucidate molecular biological and immunohistochemical characteristics of canine DP cells and determine appropriate conditions for in vitro expansion. Histological investigation revealed that the canine DP expressed biomarkers of human and rodent DP, including alkaline phosphatase (ALP) and versican. When microdissected, canine DP, but not fibroblasts, strongly expressed the DP-related genes for alkaline phosphatase, Wnt inhibitory factor 1 and lymphoid enhancer-binding factor 1, confirming successful isolation. The growth rate of isolated canine DP cells was moderate in conventional culture conditions for rodent and human DP; however, AmnioMAX-C100 complete medium allowed more efficient cultivation. Dermal papilla marker gene expression was maintained in early passage cultured DP cells, but gradually lost after the third passage. Approaches to mimic the in vivo DP environment in culture, such as supplementation of keratinocyte-conditioned medium or use of extracellular matrix-coated dishes, moderately ameliorated loss of DP gene expression in canine DP cells. It is possible that constituent factors in AmnioMAX may influence culture. These findings suggested that further refinements of culture conditions may enable DP cell expansion without impairing intrinsic properties and, importantly, demonstrated that AmnioMAX-cultured early passage canine DP cells partly maintained the biological characteristics of in vivo canine DP cells. This study provides crucial information necessary for further optimization of culture conditions of canine DP. © 2011 The Authors. Veterinary Dermatology. © 2011 ESVD and ACVD.

Characterization of primary human mammary epithelial cells isolated and propagated by conditional reprogrammed cell culture.

PubMed

Jin, Liting; Qu, Ying; Gomez, Liliana J; Chung, Stacey; Han, Bingchen; Gao, Bowen; Yue, Yong; Gong, Yiping; Liu, Xuefeng; Amersi, Farin; Dang, Catherine; Giuliano, Armando E; Cui, Xiaojiang

2018-02-20

Conditional reprogramming methods allow for the inexhaustible in vitro proliferation of primary epithelial cells from human tissue specimens. This methodology has the potential to enhance the utility of primary cell culture as a model for mammary gland research. However, few studies have systematically characterized this method in generating in vitro normal human mammary epithelial cell models. We show that cells derived from fresh normal breast tissues can be propagated and exhibit heterogeneous morphologic features. The cultures are composed of CK18, desmoglein 3, and CK19-positive luminal cells and vimentin, p63, and CK14-positive myoepithelial cells, suggesting the maintenance of in vivo heterogeneity. In addition, the cultures contain subpopulations with different CD49f and EpCAM expression profiles. When grown in 3D conditions, cells self-organize into distinct structures that express either luminal or basal cell markers. Among these structures, CK8-positive cells enclosing a lumen are capable of differentiation into milk-producing cells in the presence of lactogenic stimulus. Furthermore, our short-term cultures retain the expression of ERα, as well as its ability to respond to estrogen stimulation. We have investigated conditionally reprogrammed normal epithelial cells in terms of cell type heterogeneity, cellular marker expression, and structural arrangement in two-dimensional (2D) and three-dimensional (3D) systems. The conditional reprogramming methodology allows generation of a heterogeneous culture from normal human mammary tissue in vitro . We believe that this cell culture model will provide a valuable tool to study mammary cell function and malignant transformation.

Scaling methane oxidation: From laboratory incubation experiments to landfill cover field conditions

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

Abichou, Tarek, E-mail: abichou@eng.fsu.edu; Mahieu, Koenraad; Chanton, Jeff

2011-05-15

Evaluating field-scale methane oxidation in landfill cover soils using numerical models is gaining interest in the solid waste industry as research has made it clear that methane oxidation in the field is a complex function of climatic conditions, soil type, cover design, and incoming flux of landfill gas from the waste mass. Numerical models can account for these parameters as they change with time and space under field conditions. In this study, we developed temperature, and water content correction factors for methane oxidation parameters. We also introduced a possible correction to account for the different soil structure under field conditions.more » These parameters were defined in laboratory incubation experiments performed on homogenized soil specimens and were used to predict the actual methane oxidation rates to be expected under field conditions. Water content and temperature corrections factors were obtained for the methane oxidation rate parameter to be used when modeling methane oxidation in the field. To predict in situ measured rates of methane with the model it was necessary to set the half saturation constant of methane and oxygen, K{sub m}, to 5%, approximately five times larger than laboratory measured values. We hypothesize that this discrepancy reflects differences in soil structure between homogenized soil conditions in the lab and actual aggregated soil structure in the field. When all of these correction factors were re-introduced into the oxidation module of our model, it was able to reproduce surface emissions (as measured by static flux chambers) and percent oxidation (as measured by stable isotope techniques) within the range measured in the field.« less

Mesenchymal stem cell-conditioned medium triggers neuroinflammation and reactive species generation in organotypic cultures of rat hippocampus.

PubMed

Horn, Ana Paula; Bernardi, Andressa; Luiz Frozza, Rudimar; Grudzinski, Patrícia Bencke; Hoppe, Juliana Bender; de Souza, Luiz Fernando; Chagastelles, Pedro; de Souza Wyse, Angela Terezinha; Bernard, Elena Aida; Battastini, Ana Maria Oliveira; Campos, Maria Martha; Lenz, Guido; Nardi, Nance Beyer; Salbego, Christianne

2011-07-01

Cell therapy using bone marrow-derived mesenchymal stem cells (MSCs) seems to be a new alternative for the treatment of neurodegenerative diseases. Despite several promising results with their use, possible side effects are still unknown. In a previous work, we have shown that MSC-conditioned medium is toxic to hippocampal slice cultures and aggravates cell death induced by oxygen and glucose deprivation. In this work, we investigated whether the inflammatory response and/or reactive species formation could be involved in that toxicity. Rat organotypic hippocampal cultures were exposed for 24 h to conditioned medium from MSCs isolated from rat bone marrow. A marked glial activation was observed after exposure of cultures to MSC-conditioned medium, as evidenced by glial fibrillary acid protein (GFAP) and isolectin B(4) increase. Tumor necrosis factor-α and interleukin-6 levels were increased in the culture medium, and 2,7-dihydrodichlorofluorescein diacetate oxidation (indicating reactive species generation) and inducible nitric oxide synthase (iNOS) immunocontent were also higher after exposure of cultures to MSC-conditioned medium. Antioxidants (ascorbic acid and TROLOX(®)), N(ω)-nitro-l-arginine methyl ester hydrochloride, and anti-inflammatory drugs (indomethacin and dexamethasone) reduced cell death in hippocampal organotypic cultures after their exposure to MSC-conditioned medium. The results obtained here suggest that MSC-secreted factors trigger reactive species generation and neuroinflammation in organotypic cultures of hippocampus, introducing a note of caution in the use of these cells for neurological application.

Popular culture and the "new human condition": Catastrophe narratives and climate change

NASA Astrophysics Data System (ADS)

Bulfin, Ailise

2017-09-01

Striking popular culture images of burnt landscapes, tidal waves and ice-bound cities have the potential to dramatically and emotively convey the dangers of climate change. Given that a significant number of people derive a substantial proportion of their information on the threat of climate change, or the ;new human condition;, from popular culture works such as catastrophe movies, it is important that an investigation into the nature of the representations produced be embedded in the attempt to address the issue. What climate change-related messages may be encoded in popular films, television and novels, how are they being received, and what effects may they have? This article adopts the cultural studies perspective that popular culture gives us an important means by which to access the ;structures of feeling; that characterise a society at a particular historic juncture: the views held and emotional states experienced by significant amounts of people as evident in disparate forms of cultural production. It further adopts the related viewpoint that popular culture has an effect upon the society in which it is consumed, as well as reflecting that society's desires and concerns - although the nature of the effect may be difficult to quantify. From this position, the article puts forward a theory on the role of ecological catastrophe narratives in current popular culture, before going on to review existing critical work on ecologically-charged popular films and novels which attempts to assess their effects on their audiences. It also suggests areas for future research, such as the prevalent but little studied theme of natural and environmental disaster in late-Victorian science fiction writing. This latter area is of interest because it reveals the emergence of an ecological awareness or structure of feeling as early as the late-nineteenth century, and allows the relationship of this development to environmental policy making to be investigated because of the

Development of an economical, autonomous pHstat system for culturing phytoplankton under steady state or dynamic conditions.

PubMed

Golda, Rachel L; Golda, Mark D; Hayes, Jacqueline A; Peterson, Tawnya D; Needoba, Joseph A

2017-05-01

Laboratory investigations of physiological processes in phytoplankton require precise control of experimental conditions. Chemostats customized to control and maintain stable pH levels (pHstats) are ideally suited for investigations of the effects of pH on phytoplankton physiology, for example in context of ocean acidification. Here we designed and constructed a simple, flexible pHstat system and demonstrated its operational capabilities under laboratory culture conditions. In particular, the system is useful for simulating natural cyclic pH variability within aquatic ecosystems, such as diel fluctuations that result from metabolic activity or tidal mixing in estuaries. The pHstat system operates in two modes: (1) static/set point pH, which maintains pH at a constant level, or (2) dynamic pH, which generates regular, sinusoidal pH fluctuations by systematically varying pH according to user-defined parameters. The pHstat is self-regulating through the use of interchangeable electronically controlled reagent or gas-mediated pH-modification manifolds, both of which feature flow regulation by solenoid valves. Although effective pH control was achieved using both liquid reagent additions and gas-mediated methods, the liquid manifold exhibited tighter control (±0.03pH units) of the desired pH than the gas manifold (±0.10pH units). The precise control provided by this pHstat system, as well as its operational flexibility will facilitate studies that examine responses by marine microbiota to fluctuations in pH in aquatic ecosystems. Copyright © 2017 Elsevier B.V. All rights reserved.

Folsomia Candida--An Ideal Organism for Population Studies in the Laboratory

ERIC Educational Resources Information Center

Usher, M. B.; Stoneman, C. F.

1977-01-01

Folsomia candida is presented as an ideal organism for population studies that can be carried out cheaply and easily in school laboratory conditions. Means of identifying, obtaining, and culturing these organisms are described together with some indication of the kinds of investigations which can be performed. (Author/MA)

Value of laboratory tests in employer-sponsored health risk assessments for newly identifying health conditions: analysis of 52,270 participants.

PubMed

Kaufman, Harvey W; Williams, Fred R; Odeh, Mouneer A

2011-01-01

Employer-sponsored health risk assessments (HRA) may include laboratory tests to provide evidence of disease and disease risks for common medical conditions. We evaluated the ability of HRA-laboratory testing to provide new disease-risk information to participants. We performed a cross-sectional analysis of HRA-laboratory results for participating adult employees and their eligible spouses or their domestic partners, focusing on three common health conditions: hyperlipidemia, diabetes mellitus, and chronic kidney disease. HRA with laboratory results of 52,270 first-time participants were analyzed. Nearly all participants had access to health insurance coverage. Twenty-four percent (12,392) self-reported one or more of these medical conditions: 21.1% (11,017) self-identified as having hyperlipidemia, 4.7% (2,479) self-identified as having diabetes, and 0.7% (352) self-identified as having chronic kidney disease. Overall, 36% (n = 18,540) of participants had laboratory evidence of at least one medical condition newly identified: 30.7% (16,032) had laboratory evidence of hyperlipidemia identified, 1.9% (984) had laboratory evidence of diabetes identified, and 5.5% (2,866) had laboratory evidence of chronic kidney disease identified. Of all participants with evidence of hyperlipidemia 59% (16,030 of 27,047), were newly identified through the HRA. Among those with evidence of diabetes 28% (984 of 3,463) were newly identified. The highest rate of newly identified disease risk was for chronic kidney disease: 89% (2,866 of 3,218) of participants with evidence of this condition had not self-reported it. Men (39%) were more likely than women (33%) to have at least one newly identified condition (p<0.0001). Among men, lower levels of educational achievement were associated with modestly higher rates of newly identified disease risk (p<0.0001); the association with educational achievement among women was unclear. Even among the youngest age range (20 to 29 year olds

Evaluation of culture techniques and bacterial cultures from uroliths.

PubMed

Perry, Leigh A; Kass, Philip H; Johnson, Dee L; Ruby, Annette L; Shiraki, Ryoji; Westropp, Jodi L

2013-03-01

The association between urolithiasis and growth of bacteria in the urine or urolith has not been recently evaluated in the past 15 years, and the effects of antimicrobial administration on urolith cultures have not been reported. As well, laboratory techniques for urolith cultures have not been critically evaluated. The objectives of the current study were to 1) report bacterial isolates from uroliths and their association with signalment, urolith composition, antimicrobial use, and urine cultures and 2) evaluate laboratory techniques for urolith cultures. For the first objective, a retrospective search of bacterial isolates cultured from uroliths submitted to the laboratory as well as the signalment, urine culture results, and antimicrobial use were recorded. For the second objective, 50 urolith pairs were cultured by washing each urolith either 1or 4 times and culturing the core. Five hundred twenty canine and 168 feline uroliths were reviewed. Struvite-containing uroliths had an increased prevalence of a positive culture compared to nonstruvite-containing uroliths (P < 0.0001, odds ratio [OR] = 5.4), as did uroliths from female dogs (P < 0.0001, OR = 2.9). No significant difference between culture results and previous antimicrobial administration was found (P = 0.41). Eighteen percent of cases with negative urine cultures had positive urolith cultures. There was no significant difference in core culture results whether the urolith was washed 1 or 4 times (P = 0.07). Urolith culture outcome was not always influenced by previous antimicrobial administration, and bacterial culture of a urolith may not yield the same results as those obtained from the urine. The modified protocol, which requires less time and expense for urolith cultures, may be an acceptable alternative.

Optimization of culture conditions for gamma-aminobutyric acid production in fermented adzuki bean milk.

PubMed

Song, Hung Yi; Yu, Roch Chui

2018-01-01

γ-Aminobutyric acid (GABA), a nonprotein amino acid, is widely distributed in nature and fulfills several physiological functions. In this study, various lactic acid strains commonly used to produce fermented milk products were inoculated into adzuki bean milk for producing GABA. The high GABA producing strain was selected in further experiment to improve the GABA production utilizing culture medium optimization. The results demonstrated that adzuki bean milk inoculated with Lactobacillus rhamnosus GG increased GABA content from 0.05 mg/mL to 0.44 mg/mL after 36 hours of fermentation, which showed the greatest elevation in this study. Furthermore, the optimal cultural condition to adzuki bean milk inoculated with L. rhamnosus GG to improve the GABA content was performed using response surface methodology. The results showed that GABA content was dependent on the addition of galactose, monosodium glutamate, and pyridoxine with which the increasing ratios of GABA were 23-38%, 24-68%, and 8-36%, respectively. The optimal culture condition for GABA production of adzuki bean milk was found at the content of 1.44% galactose, 2.27% monosodium glutamate, and 0.20% pyridoxine. Under the optimal cultural condition, the amount of GABA produced in the fermented adzuki bean milk was 1.12 mg/mL, which was 22.4-fold higher than that of the unfermented adzuki bean milk (0.05 mg/100 mL). The results suggested that the optimized cultural condition of adzuki bean milk inoculated with L. rhamnosus GG can increase GABA content for consumers as a daily supplement as suggested. Copyright © 2017. Published by Elsevier B.V.

Lignolytic enzymes produced by Trametes villosa ccb176 under different culture conditions

PubMed Central

Yamanaka, Renata; Soares, Clarissa F.; Matheus, Dácio R.; Machado, Kátia M.G.

2008-01-01

The expression of the enzymatic system produced by basidiomycetous fungi, which is involved in the degradation of xenobiotics, mainly depends on culture conditions, especially of the culture medium composition. Trametes villosa is a strain with a proven biotechnological potential for the degradation of organochlorine compounds and for the decolorization of textile dyes. The influence of glucose concentration, addition of a vegetable oil-surfactant emulsion, nature of the surfactant and the presence of manganese and copper on the growth, pH and production of laccase, total peroxidase and manganese-dependent peroxidase activities were evaluated. In general, acidification of the medium was observed, with the pH reaching a value close to 3.5 within the first days of growth. Laccase was the main activity detected under the different conditions and was produced throughout the culture period of the fungus, irrespective of the growth phase. Supplementation of the medium with vegetable oil emulsified with a surfactant induced manganese-dependent peroxidase activity in T. villosa. Higher specific yields of laccase activity were obtained with the addition of copper. PMID:24031184

Laboratory-based electrical conductivity at Martian mantle conditions

NASA Astrophysics Data System (ADS)

Verhoeven, Olivier; Vacher, Pierre

2016-12-01

Information on temperature and composition of planetary mantles can be obtained from electrical conductivity profiles derived from induced magnetic field analysis. This requires a modeling of the conductivity for each mineral phase at conditions relevant to planetary interiors. Interpretation of iron-rich Martian mantle conductivity profile therefore requires a careful modeling of the conductivity of iron-bearing minerals. In this paper, we show that conduction mechanism called small polaron is the dominant conduction mechanism at temperature, water and iron content conditions relevant to Mars mantle. We then review the different measurements performed on mineral phases with various iron content. We show that, for all measurements of mineral conductivity reported so far, the effect of iron content on the activation energy governing the exponential decrease in the Arrhenius law can be modeled as the cubic square root of the iron content. We recast all laboratory results on a common generalized Arrhenius law for iron-bearing minerals, anchored on Earth's mantle values. We then use this modeling to compute a new synthetic profile of Martian mantle electrical conductivity. This new profile matches perfectly, in the depth range [100,1000] km, the electrical conductivity profile recently derived from the study of Mars Global Surveyor magnetic field measurements.

Culture Training: Validation Evidence for the Culture Assimilator.

ERIC Educational Resources Information Center

Mitchell, Terence R.; And Others

The culture assimilator, a programed self-instructional approach to culture training, is described and a series of laboratory experiments and field studies validating the culture assimilator are reviewed. These studies show that the culture assimilator is an effective method of decreasing some of the stress experienced when one works with people…

Consequences of keeping Mytilus in the laboratory as assessed by different cellular condition indices

NASA Astrophysics Data System (ADS)

Cajaraville, M. P.; Díez, G.; Marigómez, I. A.; Angulo, E.

1991-12-01

Mytilus galloprovincialis Lmk. were maintained in the laboratory for three months in a semicontinuous water flow system. Animals were fed a commercial filter-feeder food and sampled after 0, 21, 35, 49, 77, and 91 days. In order to establish whether laboratory conditions and the food used were deleterious to mussels, their health status was assessed by quantifying different histological parameters of the digestive gland tissue. It was concluded that mussels kept for more than 35 days under the described laboratory conditions showed signs of stress presumably caused by the reproductive state of the mussels investigated. The food used and the nutrition-related health status of the animals were adequate, as shown by transmission electron microscopical studies after the 91-day maintenance period. A stress response was also evoked by a 10-day starvation period, which was reflected by an increased proportion of type I and type IV digestive tubules, and a reduced “Mean Epithelial Thickness” (MET). Finally, the results demonstrate the sensitivity of quantitative histological diagnosis in comparison to subjective tubule grading procedures in the assessment of the degree of stress experienced by mussels.

"Deep-media culture condition" promoted lumen formation of endothelial cells within engineered three-dimensional tissues in vitro.

PubMed

Sekiya, Sachiko; Shimizu, Tatsuya; Yamato, Masayuki; Okano, Teruo

2011-03-01

In the field of tissue engineering, the induction of microvessels into tissues is an important task because of the need to overcome diffusion limitations of oxygen and nutrients within tissues. Powerful methods to create vessels in engineered tissues are needed for creating real living tissues. In this study, we utilized three-dimensional (3D) highly cell dense tissues fabricated by cell sheet technology. The 3D tissue constructs are close to living-cell dense tissue in vivo. Additionally, creating an endothelial cell (EC) network within tissues promoted neovascularization promptly within the tissue after transplantation in vivo. Compared to the conditions in vivo, however, common in vitro cell culture conditions provide a poor environment for creating lumens within 3D tissue constructs. Therefore, for determining adequate conditions for vascularizing engineered tissue in vitro, our 3D tissue constructs were cultured under a "deep-media culture conditions." Compared to the control conditions, the morphology of ECs showed a visibly strained cytoskeleton, and the density of lumen formation within tissues increased under hydrostatic pressure conditions. Moreover, the increasing expression of vascular endothelial cadherin in the lumens suggested that the vessels were stabilized in the stimulated tissues compared with the control. These findings suggested that deep-media culture conditions improved lumen formation in engineered tissues in vitro.

Laboratory Equipment for Investigation of Coring Under Mars-like Conditions

NASA Astrophysics Data System (ADS)

Zacny, K.; Cooper, G.

2004-12-01

To develop a suitable drill bit and set of operating conditions for Mars sample coring applications, it is essential to make tests under conditions that match those of the mission. The goal of the laboratory test program was to determine the drilling performance of diamond-impregnated bits under simulated Martian conditions, particularly those of low pressure and low temperature in a carbon dioxide atmosphere. For this purpose, drilling tests were performed in a vacuum chamber kept at a pressure of 5 torr. Prior to drilling, a rock, soil or a clay sample was cooled down to minus 80 degrees Celsius (Zacny et al, 2004). Thus, all Martian conditions, except the low gravity were simulated in the controlled environment. Input drilling parameters of interest included the weight on bit and rotational speed. These two independent variables were controlled from a PC station. The dependent variables included the bit reaction torque, the depth of the bit inside the drilled hole and the temperatures at various positions inside the drilled sample, in the center of the core as it was being cut and at the bit itself. These were acquired every second by a data acquisition system. Additional information such as the rate of penetration and the drill power were calculated after the test was completed. The weight of the rock and the bit prior to and after the test were measured to aid in evaluating the bit performance. In addition, the water saturation of the rock was measured prior to the test. Finally, the bit was viewed under the Scanning Electron Microscope and the Stereo Optical Microscope. The extent of the bit wear and its salient features were captured photographically. The results revealed that drilling or coring under Martian conditions in a water saturated rock is different in many respects from drilling on Earth. This is mainly because the Martian atmospheric pressure is in the vicinity of the pressure at the triple point of water. Thus ice, heated by contact with the

Key issues concerning environmental enrichment for laboratory-held fish species.

PubMed

Williams, T D; Readman, G D; Owen, S F

2009-04-01

An improved knowledge and understanding of the fundamental biological requirements is needed for many of the species of fish held in captivity and, without this knowledge it is difficult to determine the optimal conditions for laboratory culture. The aim of this paper is to review the key issues concerning environmental enrichment for laboratory-held fish species and identify where improvements are required. It provides background information on environmental enrichment, describes enrichment techniques currently used in aquatic ecotoxicology studies, identifies potential restrictions in their use and discusses why more detailed and species-specific guidance is needed.

Nonlinear Dielectric Properties of Yeast Cells Cultured in Different Environmental Conditions

NASA Astrophysics Data System (ADS)

Kawanishi, Gomon; Fukuda, Naoki; Muraji, Masafumi

The harmonics of the electric current through yeast suspensions, the nonlinear dielectric properties of yeast cells, have particular patterns according to the biological activity of the cells and the measurement of these patterns is a technique for determining the activity of living cells. The concentration of glucose and oxygen in yeast culture medium influences the manifestation of fermentation or respiration of yeast cells. Measurements were made with yeast cells (Saccharomyces cerevisiae) cultured aerobically and anaerobically in sufficient glucose concentration, aerobic fermentation and anaerobic fermentation, and aerobically in limited glucose concentration, respiration. The results showed that the harmonics were barely apparent for yeast cells in aerobic fermentation and respiratory; however, cells in the anaerobic fermentation displayed substantial third and fifth harmonics. We can say that environmental condition affects the yeast cells' nonlinear properties, from another viewpoint, the measurements of the nonlinear properties are available to determine the activity of yeast cells adjusted to the conditions of their cultivation.

Masters of their conditions: at the crossroads of health, culture and performance.

PubMed

Arpin, Jacques

2003-09-01

Achieving one's cultural or clinical identity is a matter of performance. The performing body is made through a series of apprenticeships that lead to mastery and then transmission. Performance, like culture and medicine, has its systems of learning, i.e. in the professions of the performing arts. This article examines the methodology of theater anthropology and other aspects of performance studies. Applications to intercultural work are described that make it possible for patient and healer to collaborate to master the condition.

Culturing primary mouse pancreatic ductal cells.

PubMed

Reichert, Maximilian; Rhim, Andrew D; Rustgi, Anil K

2015-06-01

The most common subtype of pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC). PDAC resembles ductal cells morphologically. To study pancreatic ductal cell (PDC) and pancreatic intraepithelial neoplasia (PanIN)/PDAC biology, it is essential to have reliable in vitro culture conditions. Here we describe a methodology to isolate, culture, and passage PDCs and duct-like cells from the mouse pancreas. It can be used to isolate cells from genetically engineered mouse models (GEMMs), providing a valuable tool to study disease models in vitro to complement in vivo findings. The culture conditions allow epithelial cells to outgrow fibroblast and other "contaminating" cell types within a few passages. However, the resulting cultures, although mostly epithelial, are not completely devoid of fibroblasts. Regardless, this protocol provides guidelines for a robust in vitro culture system to isolate, maintain, and expand primary pancreatic ductal epithelial cells. It can be applied to virtually all GEMMs of pancreatic disease and other diseases and cancers that arise from ductal structures. Because most carcinomas resemble ductal structures, this protocol has utility in the study of other cancers in addition to PDAC, such as breast and prostate cancers. © 2015 Cold Spring Harbor Laboratory Press.

Use of benthic prey by salmonids under turbid conditions in a laboratory stream

Treesearch

Bret C. Harvey; Jason L. White

2008-01-01

The negative effect of turbidity on the reactive distance of salmonids has been well established. However, determining the consequences of this relationship for overall feeding success remains problematic, as successful foraging by salmonids across a broad range in turbidity has been observed under a variety of conditions. Previous laboratory and field observations...

Fast screening of Bifidobacterium longum sublethal stress conditions in a novel two-stage continuous culture strategy.

PubMed

Mozzetti, V; Grattepanche, F; Berger, B; Rezzonico, E; Arigoni, F; Lacroix, C

2013-06-01

A central issue in the application of probiotics as food additives is their fastidious production and their sensitivity to many environmental stresses. The importance of inducible cell-protective mechanisms triggered by application of sublethal stresses for survival under stress conditions has been demonstrated. Continuous cultures could be a suitable and more efficient method to test stress factors on one culture instead of several repeated batch cultures. In this study, the application of a two-stage continuous culture of Bifidobacterium longum NCC2705 was investigated. The first reactor was operated under fixed conditions at 37 °C and pH 6.0 and used to produce cells with controlled physiology, mimicking cells in the late exponential growth phase. Stress pretreatment combinations of pH (6.0, 5.0 and 4.0), temperature (37, 45 and 47 °C) and NaCl (0, 5 and 10%) were tested in the second reactor. Of all tested combinations, only those of pH 4.0 significantly decreased cell viability in the second reactor compared to control conditions (37 °C, pH 6.0, 0% NaCl) and, therefore, could not be considered as sublethal stresses. Pretreatments with 5 or 10% NaCl had a negative effect on cell viability after gastric lethal stress. A significant improvement in cell resistance to heat lethal stress (56 °C, 5 min) was observed for cells pretreated at 47 °C. In contrast, heat pretreatment negatively affected cell viability after freeze drying and osmotic lethal stresses. The two-stage continuous culture allowed for efficient screening of several stress pretreatments during the same experiment with up to four different conditions tested per day. Optimal sublethal stress conditions can also be applied for producing cells with traditional batch cultures.

An integrated system for synchronous culture of animal cells under controlled conditions.

PubMed

Mendoza-Pérez, Elena; Hernández, Vanessa; Palomares, Laura A; Serrato, José A

2016-01-01

The cell cycle has fundamental effects on cell cultures and their products. Tools to synchronize cultured cells allow the study of cellular physiology and metabolism at particular cell cycle phases. However, cells are most often arrested by methods that alter their homeostasis and are then cultivated in poorly controlled environments. Cell behavior could then be affected by the synchronization method and culture conditions used, and not just by the particular cell cycle phase under study. Moreover, only a few viable cells are recovered. Here, we designed an integrated system where a large number of cells from a controlled bioreactor culture is separated by centrifugal elutriation at high viabilities. In contrast to current elutriation methods, cells are injected directly from a bioreactor into an injection loop, allowing the introduction of a large number of cells into the separation chamber without stressful centrifugation. A low pulsation peristaltic pump increases the stability of the elutriation chamber. Using this approach, a large number of healthy cells at each cell cycle phase were obtained, allowing their direct inoculation into fully instrumented bioreactors. Hybridoma cells synchronized and cultured in this system behaved as expected for a synchronous culture.

Rapid Evolution of Culture-Impaired Bacteria During Adaptation to Biofilm Growth

PubMed Central

Penterman, Jon; Nguyen, Dao; Anderson, Erin; Staudinger, Benjamin J.; Greenberg, Everett P.; Lam, Joseph S.; Singh, Pradeep K.

2014-01-01

Summary Biofilm growth increases the fitness of bacteria in harsh conditions. However, bacteria from clinical and environmental biofilms can exhibit impaired growth in culture, even when the species involved are readily cultureable, and permissive conditions are used. Here we show that culture-impaired variants of Pseudomonas aeruginosa rapidly and abundantly evolve in laboratory biofilms. The culture-impaired phenotype is caused by mutations that alter the outer-membrane lipopolysaccharide structure. Within biofilms, the lipopolysaccharide mutations markedly increase bacterial fitness. However, outside the protected biofilm environment, the mutations sensitize the variants to killing by a self-produced antimicrobial agent. Thus, a biofilm-mediated adaptation produces a stark fitness trade off that compromises bacterial survival in culture. Trade offs like this could limit the ability of bacteria to transition between biofilm growth and the free-living state, and produce bacterial populations that escape detection by culture-based sampling. PMID:24412364

Comparison of culture media for the laboratory diagnosis of chancroid.

PubMed

Pillay, A; Hoosen, A A; Loykissoonlal, D; Glock, C; Odhav, B; Sturm, A W

1998-11-01

Seven different agar-based media were compared to determine the optimal set of culture media for primary isolation of Haemophilus ducreyi. Also, a new method for sampling genital ulcers -- with a disposable sterile plastic loop -- and processing specimens that provides a standardised inoculum for culture of H. ducreyi on various media is described. A total of 202 patients with genital ulcer disease was enrolled in this study. A sterile swab or plastic loop was used to sample the base of the ulcers and ulcer material was suspended in sterile phosphate-buffered saline. A 100-microl sample of this suspension was mixed with an equal volume of tryptic soy broth containing IsoVitaleX and centrifuged for 1 min. This suspension was used to inoculate the different media. Plates were incubated at 33 degrees C in micro-aerophilic conditions and examined for growth of H. ducreyi after 48 h. Of the 202 specimens, 77 (38.1%) were culture positive for H. ducreyi. None of the agar bases supported the growth of all H. ducreyi strains. Based on this observation, we recommend the universal use of Mueller-Hinton agar base supplemented with chocolate horse blood and IsovitaleX (MH-HBC) and Columbia agar base supplemented with bovine haemoglobin, activated charcoal, fetal calf serum and IsovitaleX (C-HgCh) to enable comparison of prevalence figures for chancroid. In addition, the novel sampling technique described in this study eliminates sampling bias normally associated with genital ulcer specimens.

The influence of culture conditions on the identification of Mycobacterium species by MALDI-TOF MS profiling.

PubMed

Balážová, Tereza; Makovcová, Jitka; Šedo, Ondrej; Slaný, Michal; Faldyna, Martin; Zdráhal, Zbyněk

2014-04-01

Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) represents a simple reliable approach for rapid bacterial identification based on specific peptide/protein fingerprints. However, cell-wall characteristics of mycobacterial species, and their well known stability, complicate MALDI-TOF MS profiling analysis. In this study, we tested two recently published protocols for inactivation and disruption of mycobacteria, and we also examined the influence of different culture conditions (four culture media and five cultivation times) on mass spectral quality and the discriminatory power of the method. We found a significant influence of sample pretreatment method and culture medium on species identification and differentiation for a total of 10 strains belonging to Mycobacterium phlei and Mycobacterium smegmatis. Optimum culture conditions yielding the highest identification success rate against the BioTyper database (Bruker Daltonics) and permitting the possibility of automatic acquisition of mass spectra were found to be distinct for the two mycobacterial species examined. Similarly, individual changes in growth conditions had diverse effects on the two species. For these reasons, thorough control over cultivation conditions should always be employed to maximize the performance and discriminatory power of MALDI-TOF MS profiling, and cultivation conditions must be optimized separately for individual groups of mycobacterial species/strains. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

GROSS N TRANSFORMATION RATES AND MICROBIAL POPULATION DYNAMICS UNDER FIELD AND LABORATORY CONDITIONS FROM TWO DIFFERENT ECOSYSTEMS

EPA Science Inventory

Change of soil and environmental conditions can influence microbial activities and subsequent soil nitrogen (N) transformation processes. The objective of this study was to compare gross N transformation rates between field and laboratory incubation conditions using an old-field...

Good Cell Culture Practice for stem cells and stem-cell-derived models.

PubMed

Pamies, David; Bal-Price, Anna; Simeonov, Anton; Tagle, Danilo; Allen, Dave; Gerhold, David; Yin, Dezhong; Pistollato, Francesca; Inutsuka, Takashi; Sullivan, Kristie; Stacey, Glyn; Salem, Harry; Leist, Marcel; Daneshian, Mardas; Vemuri, Mohan C; McFarland, Richard; Coecke, Sandra; Fitzpatrick, Suzanne C; Lakshmipathy, Uma; Mack, Amanda; Wang, Wen Bo; Yamazaki, Daiju; Sekino, Yuko; Kanda, Yasunari; Smirnova, Lena; Hartung, Thomas

2017-01-01

The first guidance on Good Cell Culture Practice (GCCP) dates back to 2005. This document expands this to include aspects of quality assurance for in vitro cell culture focusing on the increasingly diverse cell types and culture formats used in research, product development, testing and manufacture of biotechnology products and cell-based medicines. It provides a set of basic principles of best practice that can be used in training new personnel, reviewing and improving local procedures, and helping to assure standard practices and conditions for the comparison of data between laboratories and experimentation performed at different times. This includes recommendations for the documentation and reporting of culture conditions. It is intended as guidance to facilitate the generation of reliable data from cell culture systems, and is not intended to conflict with local or higher level legislation or regulatory requirements. It may not be possible to meet all recommendations in this guidance for practical, legal or other reasons. However, when it is necessary to divert from the principles of GCCP, the risk of decreasing the quality of work and the safety of laboratory staff should be addressed and any conclusions or alternative approaches justified. This workshop report is considered a first step toward a revised GCCP 2.0.

How to isolate, identify and determine antimicrobial susceptibility of anaerobic bacteria in routine laboratories?

PubMed

Nagy, E; Boyanova, L; Justesen, U S

2018-02-17

There has been increased interest in the study of anaerobic bacteria that cause human infection during the past decade. Many new genera and species have been described using 16S rRNA gene sequencing of clinical isolates obtained from different infection sites with commercially available special culture media to support the growth of anaerobes. Several systems, such as anaerobic pouches, boxes, jars and chambers provide suitable anaerobic culture conditions to isolate even strict anaerobic bacteria successfully from clinical specimens. Beside the classical, time-consuming identification methods and automated biochemical tests, the use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has revolutionized identification of even unusual and slow-growing anaerobes directly from culture plates, providing the possibility of providing timely information about anaerobic infections. The aim of this review article is to present methods for routine laboratories, which carry out anaerobic diagnostics on different levels. Relevant data from the literature mostly published during the last 7 years are encompassed and discussed. The review involves topics on the anaerobes that are members of the commensal microbiota and their role causing infection, the key requirements for collection and transport of specimens, processing of specimens in the laboratory, incubation techniques, identification and antimicrobial susceptibility testing of anaerobic bacteria. Advantages, drawbacks and specific benefits of the methods are highlighted. The present review aims to update and improve anaerobic microbiology in laboratories with optimal conditions as well as encourage its routine implementation in laboratories with restricted resources. Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Comparison of cyanobacterial microcystin synthetase (mcy) E gene transcript levels, mcy E gene copies, and biomass as indicators of microcystin risk under laboratory and field conditions.

PubMed

Ngwa, Felexce F; Madramootoo, Chandra A; Jabaji, Suha

2014-08-01

Increased incidences of mixed assemblages of microcystin-producing and nonproducing cyanobacterial strains in freshwater bodies necessitate development of reliable proxies for cyanotoxin risk assessment. Detection of microcystin biosynthetic genes in water blooms of cyanobacteria is generally indicative of the presence of potentially toxic cyanobacterial strains. Although much effort has been devoted toward elucidating the microcystin biosynthesis mechanisms in many cyanobacteria genera, little is known about the impacts of co-occurring cyanobacteria on cellular growth, mcy gene expression, or mcy gene copy distribution. The present study utilized conventional microscopy, qPCR assays, and enzyme-linked immunosorbent assay to study how competition between microcystin-producing Microcystis aeruginosa CPCC 299 and Planktothrix agardhii NIVA-CYA 126 impacts mcyE gene expression, mcyE gene copies, and microcystin concentration under controlled laboratory conditions. Furthermore, analyses of environmental water samples from the Missisquoi Bay, Quebec, enabled us to determine how the various potential toxigenic cyanobacterial biomass proxies correlated with cellular microcystin concentrations in a freshwater lake. Results from our laboratory study indicated significant downregulation of mcyE gene expression in mixed cultures of M. aeruginosa plus P. agardhii on most sampling days in agreement with depressed growth recorded in the mixed cultures, suggesting that interaction between the two species probably resulted in suppressed growth and mcyE gene expression in the mixed cultures. Furthermore, although mcyE gene copies and McyE transcripts were detected in all laboratory and field samples with measureable microcystin levels, only mcyE gene copies showed significant positive correlations (R(2) > 0.7) with microcystin concentrations, while McyE transcript levels did not. These results suggest that mcyE gene copies are better indicators of potential risks from microcystins

Esophageal culture

MedlinePlus

... page: //medlineplus.gov/ency/article/003764.htm Esophageal culture To use the sharing features on this page, please enable JavaScript. Esophageal culture is a laboratory test that checks for infection- ...

Endocervical culture

MedlinePlus

... page: //medlineplus.gov/ency/article/003754.htm Endocervical culture To use the sharing features on this page, please enable JavaScript. Endocervical culture is a laboratory test that helps identify infection ...

Comparison between point-of-care dermatophyte test medium and mycology laboratory culture for diagnosis of dermatophytosis in dogs and cats.

PubMed

Kaufmann, Ronnie; Blum, Shlomo E; Elad, Daniel; Zur, Gila

2016-08-01

Point-of-care Dermatophyte Test Medium (PoC-DTM) is a diagnostic procedure to rule in/rule out dermatophytosis in veterinary clinics. To evaluate the performance of PoC-DTM in the clinic compared to DTM plate culture in a mycology laboratory and to compare results obtained by general practitioners and referral clinicians. Hair samples were collected from 47 cats and 54 dogs with suspected dermatophytosis and from nine healthy controls (seven cats and two dogs). This was a multicentre blinded study. In one group (65 suspected cases, 9 healthy controls), PoC-DTM results were evaluated by clinicians in a referral clinic (SP group) who examined the colony morphology macroscopically and microscopically. In the other group (36 suspected cases) PoC-DTM results were evaluated by clinicians from general practice for colour change only, with no macroscopic or microscopic examination (GP group). All hair samples were also cultured on DTM plates in a mycology laboratory. Laboratory culture was considered the gold standard for comparison. Agreements between tests were 97% (two false positive; κ = 0.839) and 80.6% (five false positives and two false negatives; κ = 0.466) in the SP and GP groups, respectively. This difference between groups was significant (P = 0.024). When applying macroscopic and microscopic evaluation of the colony, PoC-DTM is accurate for diagnosing dermatophytes with only a 3% chance of error. However, when macroscopic and microscopic examination is not included there is significant (19.4%) chance for an incorrect diagnosis. © 2016 ESVD and ACVD.

The widely distributed hard tick, Haemaphysalis longicornis, can retain canine parvovirus, but not be infected in laboratory condition

PubMed Central

MORI, Hiroyuki; TANAKA, Tetsuya; MOCHIZUKI, Masami

2014-01-01

ABSTRACT. Ticks are known to transmit various pathogens, radically threatening humans and animals. Despite the close contact between ticks and viruses, our understanding on their interaction and biology is still lacking. The aim of this study was to experimentally assess the interaction between canine parvovirus (CPV) and a widely distributed hard tick, Haemaphysalis longicornis, in laboratory condition. After inoculation of CPV into the hemocoel of the ticks, polymerase chain reaction assay revealed that CPV persisted in inoculated unfed adult female ticks for 28 days. Canine parvovirus was recovered from the inoculated ticks using a cell culture, indicating that the virus retained intact in the ticks after inoculation, but significant positive reaction indicating virus infection was not detected in the tick organs by immunofluorescence antibody test using a monoclonal antibody. In the case of ticks inoculated with feline leukemia virus, the virus had shorter persistence in the ticks compared to CPV. These findings provide significant important information on the characteristic interaction of tick with non-tick-borne virus. PMID:25650060

Scientific equity: experiments in laboratory education in Ghana.

PubMed

Osseo-Asare, Abena Dove

2013-12-01

During the 1960s the Ministry of Education in Ghana created a network of school laboratories to increase scientific literacy among young citizens. The ministry stocked these "Science Centres" with imported beakers, Bunsen burners, and books. Education officials and university scientists worked with teachers to create lesson plans on water, air, plants, and other topics. The government hoped that scientifically minded schoolchildren would be better prepared to staff the industries of the future. The adoption of laboratory norms represented a desire for scientific equity, rather than a condition of cultural mimicry. Interviews with ministry officials and science educators, alongside letters and reports, indicate how students and teachers appropriated the laboratories in the small West African nation. Their experiences in mobilizing resources from across Ghana and around the world provide a metaphor for ongoing efforts to establish access to scientific goods in Africa.

Effects of culture conditions on monosaccharide composition of Ganoderma lucidum exopolysaccharide and on activities of related enzymes.

PubMed

Peng, Lin; Qiao, Shuangkui; Xu, Zhenghong; Guan, Feng; Ding, Zhongyang; Gu, Zhenghua; Zhang, Liang; Shi, Guiyang

2015-11-20

We investigated the relationship between monosaccharide composition of Ganoderma lucidum exopolysaccharide (EPS) and activities of EPS synthesis enzymes under various culture temperatures and initial pH values. The mole percentages of three major EPS monosaccharides, glucose, galactose and mannose, varied depending on culture conditions and the resulting EPS displayed differing anti-tumor activities. In nine tested enzymes, higher enzyme activities were correlated with higher temperature and lower initial pH. Altered mole percentages of galactose and mannose under various culture conditions were associated with activities of α-phosphoglucomutase (PGM) and phosphoglucose isomerase (PGI), respectively, and that of mannose was also associated with phosphomannose isomerase (PMI) activity only under various pH. Our findings suggest that mole percentages of G. lucidum EPS monosaccharides can be manipulated by changes of culture conditions that affect enzyme activities, and that novel fermentation strategies based on this approach may enhance production and biological activity of EPS. Copyright © 2015 Elsevier Ltd. All rights reserved.

System for measuring oxygen consumption rates of mammalian cells in static culture under hypoxic conditions.

PubMed

Kagawa, Yuki; Miyahara, Hirotaka; Ota, Yuri; Tsuneda, Satoshi

2016-01-01

Estimating the oxygen consumption rates (OCRs) of mammalian cells in hypoxic environments is essential for designing and developing a three-dimensional (3-D) cell culture system. However, OCR measurements under hypoxic conditions are infrequently reported in the literature. Here, we developed a system for measuring OCRs at low oxygen levels. The system injects nitrogen gas into the environment and measures the oxygen concentration by an optical oxygen microsensor that consumes no oxygen. The developed system was applied to HepG2 cells in static culture. Specifically, we measured the spatial profiles of the local dissolved oxygen concentration in the medium, then estimated the OCRs of the cells. The OCRs, and also the pericellular oxygen concentrations, decreased nonlinearly as the oxygen partial pressure in the environment decreased from 19% to 1%. The OCRs also depended on the culture period and the matrix used for coating the dish surface. Using this system, we can precisely estimate the OCRs of various cell types under environments that mimic 3-D culture conditions, contributing crucial data for an efficient 3-D culture system design. © 2015 American Institute of Chemical Engineers.

Routine sputum culture

MedlinePlus

Sputum culture ... There, it is placed in a special dish (culture). It is then watched to see if bacteria ... Elsevier; 2018:chap 36. Chernecky CC, Berger BJ. Culture, routine. In: Chernecky CC, Berger BJ, eds. Laboratory ...

On triatomines, cockroaches and haemolymphagy under laboratory conditions: new discoveries

PubMed Central

Durán, Pamela; Siñani, Edda; Depickère, Stéphanie

2016-01-01

For a long time, haematophagy was considered an obligate condition for triatomines (Hemiptera: Reduviidae) to complete their life cycle. Today, the ability to use haemolymphagy is suggested to represent an important survival strategy for some species, especially those in genus Belminus. As Eratyrus mucronatus and Triatoma boliviana are found with cockroaches in the Blaberinae subfamily in Bolivia, their developmental cycle from egg to adult under a “cockroach diet” was studied. The results suggested that having only cockroach haemolymph as a food source compromised development cycle completion in both species. Compared to a “mouse diet”, the cockroach diet increased: (i) the mortality at each nymphal instar; (ii) the number of feedings needed to molt; (iii) the volume of the maximum food intake; and (iv) the time needed to molt. In conclusion, haemolymph could effectively support survival in the field in both species. Nevertheless, under laboratory conditions, the use of haemolymphagy as a survival strategy in the first developmental stages of these species was not supported, as their mortality was very high. Finally, when Triatoma infestans, Rhodnius stali and Panstrongylus rufotuberculatus species were reared on a cockroach diet under similar conditions, all died rather than feeding on cockroaches. These results are discussed in the context of the ecology of each species. PMID:27706376

Optimization of culturing conditions of recombined Escherichia coli to produce umami octopeptide-containing protein.

PubMed

Zhang, Yin; Wei, Xiong; Lu, Zhou; Pan, Zhongli; Gou, Xinhua; Venkitasamy, Chandrasekar; Guo, Siya; Zhao, Liming

2017-07-15

Using synthesized peptides to verify the taste of natural peptides was probably the leading cause for tasting disputes regarding umami peptides. A novel method was developed to prepare the natural peptide which could be used to verify the taste of umami peptide. A controversial octopeptide was selected and gene engineering was used to structure its Escherichia coli. expressing vector. A response surface method was adopted to optimize the expression conditions of the recombinant protein. The results of SDS-PAGE for the recombinant protein indicated that the recombinant expression system was successfully structured. The fitting results of the response surface experiment showed that the OD 600 value was the key factor which influenced the expression of the recombinant protein. The optimal culturing process conditions predicted with the fitting model were an OD 600 value of 0.5, an IPTG concentration of 0.6mM, a culturing temperature of 28.75°C and a culturing time of 5h. Copyright © 2017 Elsevier Ltd. All rights reserved.

Relative efficiencies of two air sampling methods and three culture conditions for the assessment of airborne culturable fungi in a poultry farmhouse in France.

PubMed

Nieguitsila, Adélaïde; Arné, Pascal; Durand, Benoît; Deville, Manjula; Benoît-Valiergue, Hélène; Chermette, René; Cottenot-Latouche, Sophie; Guillot, Jacques

2011-02-01

Fungal elements represent a significant part of the biological contaminants that could be detected in the air of animal facilities. The aim of this study was to assess the relative efficiencies of two air sampling methods and three culture conditions for the quantification of airborne culturable fungi in a poultry farmhouse in France. Air samples were collected every week throughout a 15-week period. Two devices were simultaneously used-a rotative cup air sampler (CIP 10-M, Arelco, France) and an air sampler based on filtration (AirPort MD8, Sartorius, Germany). Culture of airborne viable fungi was performed on malt extract agar (ME) and dichloran glycerol-18 (DG18) at 25 or 37°C. CIP 10-M and AirPort MD8 were shown to display comparable performances but significant differences were observed between culture conditions for Aspergillus spp. (p<0.01), Scopulariopsis spp. (p=0.02) and unidentified molds (p<0.01). Copyright © 2010 Elsevier Inc. All rights reserved.

Method for Producing Non-Neoplastic, Three Dimensional, Mammalian Tissue and Cell Aggregates Under Microgravity Culture Conditions and the Products Produced Therefrom

NASA Technical Reports Server (NTRS)

Goodwin, Thomas J. (Inventor); Wolf, David A. (Inventor); Spaulding, Glenn F. (Inventor); Prewett, Tracey L. (Inventor)

1996-01-01

Normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural, and blood tissue. The cells are grown in vitro under microgravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel.

Cultured bovine granulosa cells rapidly lose important features of their identity and functionality but partially recover under long-term culture conditions.

PubMed

Yenuganti, Vengala Rao; Vanselow, Jens

2017-05-01

Cell culture models are essential for the detailed study of molecular processes. We analyze the dynamics of changes in a culture model of bovine granulosa cells. The cells were cultured for up to 8 days and analyzed for steroid production and gene expression. According to the expression of the marker genes CDH1, CDH2 and VIM, the cells maintained their mesenchymal character throughout the time of culture. In contrast, the levels of functionally important transcripts and of estradiol and progesterone production were rapidly down-regulated but showed a substantial up-regulation from day 4. FOXL2, a marker for granulosa cell identity, was also rapidly down-regulated after plating but completely recovered towards the end of culture. In contrast, expression of the Sertoli cell marker SOX9 and the lesion/inflammation marker PTGS2 increased during the first 2 days after plating but gradually decreased later on. We conclude that only long-term culture conditions (>4 days) allow the cells to recover from plating stress and to re-acquire characteristic granulosa cell features.

Pretreatments, conditioned medium and co-culture increase the incidence of somatic embryogenesis of different Cichorium species

PubMed Central

Couillerot, Jean-Paul; Windels, David; Vazquez, Franck; Michalski, Jean-Claude; Hilbert, Jean-Louis; Blervacq, Anne-Sophie

2012-01-01

Somatic embryogenesis (SE) in Cichorium involves dedifferentiation and redifferentiation of single cells and can be induced by specific in vitro culture conditions. We have tested the effect of various treatments on the incidence of SE (ISE) of an interspecific embryogenic hybrid (C. endivia x C. intybus) and of different commercial chicories (C. endivia and C. intybus) that are typically recalcitrant to SE in standard culture conditions. We found that the ISE of the hybrid is significantly increased by pretreatment of tissues by submersion in solutions of glycerol, abscisic acid, spermine, putrescine or of combinations of these compounds. Interestingly, the most efficient of these pretreatments also had an unexpectedly high effect on the ISE of the C. intybus cultivars. The ISE of the hybrid and of the commercial chicories were increased when explants were co-cultured with highly embryogenic chicory explants or when they were cultured in conditioned medium. These observations established that unidentified SE-promoting factors are released in the culture medium. HPLC analyses of secreted Arabino-Galactan Proteins (AGPs), which are known to stimulate SE, did not allow identifying a fraction containing differentially abundant AGP candidates. However, pointing to their role in promoting SE, we found that the hybrid had a drastically higher ISE when amino sugars and L-Proline, the putative precursors of secreted AGPs, were both added to the medium. PMID:22301978

Comparison of the efficiency of transplantation of bone marrow multipotent mesenchymal stromal cells cultured under normoxic and hypoxic conditions and their conditioned media on the model of acute lung injury.

PubMed

Chailakhyan, R K; Aver'yanov, A V; Zabozlaev, F G; Sobolev, P A; Sorokina, A V; Akul'shin, D A; Gerasimov, Yu V

2014-05-01

The therapeutic efficiency of intravenous injection of rat bone marrow multipotent mesenchymal stromal cells grown under conditions of normoxia and hypoxia (3% O2) and conditioned media from these cultures were compared on the rat model of acute lung injury induced by intraperitoneal injection of lipopolysaccharide. The best therapeutic efficiency was demonstrated by cells grown under hypoxic conditions. The effect of conditioned media was less pronounced and did not depend on the culturing conditions.

A sensible technique to detect mollicutes impurities in human cells cultured in GMP condition.

PubMed

Ugolotti, Elisabetta; Vanni, Irene

2014-01-01

In therapeutic trials the use of manipulated cell cultures for clinical applications is often required. Mollicutes microorganism contamination of tissue cultures is a major problem because it can determine various and severe alterations in cellular function. Thus methods able to detect and trace cell cultures with Mollicutes contamination are needed in the monitoring of cells grown under good manufacturing practice conditions, and cell lines in continuous culture must be tested at regular intervals. We here describe a multiplex quantitative polymerase chain reaction assay able to detect contaminant Mollicutes species in a single-tube reaction through analysis of 16S-23S rRNA intergenic spacer regions and Tuf and P1 cytoadhesin genes. The method shows a sensitivity, specificity, and robustness comparable with the culture and the indicator cell culture as required by the European Pharmacopoeia guidelines and was validated following International Conference on Harmonization guidelines and Food and Drug Administration requirements.

Optimization of cultural conditions for conversion of glycerol to ethanol by Enterobacter aerogenes S012

PubMed Central

2013-01-01

The aim of this research is to optimize the cultural conditions for the conversion of glycerol to ethanol by Enterobacter aerogenes S012. Taguchi method was used to screen the cultural conditions based on their signal to noise ratio (SN). Temperature (°C), agitation speed (rpm) and time (h) were found to have the highest influence on both glycerol utilization and ethanol production by the organism while pH had the lowest. Full factorial design, statistical analysis, and regression model equation were used to optimize the selected cultural parameters for maximum ethanol production. The result showed that fermentation at 38°C and 200 rpm for 48 h would be ideal for the bacteria to produce maximum amount of ethanol from glycerol. At these optimum conditions, ethanol production, yield and productivity were 25.4 g/l, 0.53 g/l/h, and 1.12 mol/mol-glycerol, repectively. Ethanol production increased to 26.5 g/l while yield and productivity decreased to 1.04 mol/mol-glycerol and 0.37 g/l/h, respectively, after 72 h. Analysis of the fermentation products was performed using HPLC, while anaerobic condition was created by purging the fermentation vessel with nitrogen gas. PMID:23388539

Cultural Resource Investigations for the Resumption of Transient Testing of Nuclear Fuels and Material at the Idaho National Laboratory

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

Pace, Brenda R.; Williams, Julie B.

2013-11-01

The U. S. Department of Energy (DOE) has a need to test nuclear fuels under conditions that subject them to short bursts of intense, high-power radiation called ‘transient testing’ in order to gain important information necessary for licensing new nuclear fuels for use in U.S. nuclear power plants, for developing information to help improve current nuclear power plant performance and sustainability, for improving the affordability of new generation reactors, for developing recyclable nuclear fuels, and for developing fuels that inhibit any repurposing into nuclear weapons. To meet this mission need, DOE is considering alternatives for re-use and modification of existingmore » nuclear reactor facilities to support a renewed transient testing program. One alternative under consideration involves restarting the Transient Reactor Test (TREAT) reactor located at the Materials and Fuels Complex (MFC) on the Idaho National Laboratory (INL) site in southeastern Idaho. This report summarizes cultural resource investigations conducted by the INL Cultural Resource Management Office in 2013 to support environmental review of activities associated with restarting the TREAT reactor at the INL. These investigations were completed in order to identify and assess the significance of cultural resources within areas of potential effect associated with the proposed action and determine if the TREAT alternative would affect significant cultural resources or historic properties that are eligible for nomination to the National Register of Historic Places. No archaeological resources were identified in the direct area of potential effects for the project, but four of the buildings proposed for modifications are evaluated as historic properties, potentially eligible for nomination to the National Register of Historic Places. This includes the TREAT reactor (building #), control building (building #), guardhouse (building #), and warehouse (building #). The proposed re-use of these

Culture media from hypoxia conditioned endothelial cells protect human intestinal cells from hypoxia/reoxygenation injury.

PubMed

Hummitzsch, Lars; Zitta, Karina; Bein, Berthold; Steinfath, Markus; Albrecht, Martin

2014-03-10

Remote ischemic preconditioning (RIPC) is a phenomenon, whereby short episodes of non-lethal ischemia to an organ or tissue exert protection against ischemia/reperfusion injury in a distant organ. However, there is still an apparent lack of knowledge concerning the RIPC-mediated mechanisms within the target organ and the released factors. Here we established a human cell culture model to investigate cellular and molecular effects of RIPC and to identify factors responsible for RIPC-mediated intestinal protection. Human umbilical vein cells (HUVEC) were exposed to repeated episodes of hypoxia (3 × 15 min) and conditioned culture media (CM) were collected after 24h. Human intestinal cells (CaCo-2) were cultured with or without CM and subjected to 90 min of hypoxia/reoxygenation injury. Reverse transcription-polymerase chain reaction, Western blotting, gelatin zymography, hydrogen peroxide measurements and lactate dehydrogenase (LDH) assays were performed. In HUVEC cultures hypoxic conditioning did not influence the profile of secreted proteins but led to an increased gelatinase activity (P<0.05) in CM. In CaCo-2 cultures 90 min of hypoxia/reoxygenation resulted in morphological signs of cell damage, increased LDH levels (P<0.001) and elevated levels of hydrogen peroxide (P<0.01). Incubation of CaCo-2 cells with CM reduced the hypoxia-induced signs of cell damage and LDH release (P<0.01) and abrogated the hypoxia-induced increase of hydrogen peroxide. These events were associated with an enhanced phosphorylation status of the prosurvival kinase Erk1/2 (P<0.05) but not Akt and STAT-5. Taken together, CM of hypoxia conditioned endothelial cells protect human intestinal cells from hypoxia/reoxygenation injury. The established culture model may help to unravel RIPC-mediated cellular events and to identify molecules released by RIPC. Copyright © 2014 Elsevier Inc. All rights reserved.

Peritoneal fluid culture

MedlinePlus

Culture - peritoneal fluid ... sent to the laboratory for Gram stain and culture. The sample is checked to see if bacteria ... The peritoneal fluid culture may be negative, even if you have ... diagnosis of peritonitis is based on other factors, in addition ...

The standardized fish bioassay procedure for detecting and culturing actively toxic Pfiesteria, used by two reference laboratories for atlantic and gulf coast states.

PubMed Central

Burkholder, J M; Marshall, H G; Glasgow, H B; Seaborn, D W; Deamer-Melia, N J

2001-01-01

In the absence of purified standards of toxins from Pfiesteria species, appropriately conducted fish bioassays are the "gold standard" that must be used to detect toxic strains of Pfiesteria spp. from natural estuarine water or sediment samples and to culture actively toxic Pfiesteria. In this article, we describe the standardized steps of our fish bioassay as an abbreviated term for a procedure that includes two sets of trials with fish, following the Henle-Koch postulates modified for toxic rather than infectious agents. This procedure was developed in 1991, and has been refined over more than 12 years of experience in research with toxic Pfiesteria. The steps involve isolating toxic strains of Pfiesteria (or other potentially, as-yet-undetected, toxic Pfiesteria or Pfiesteria-like species) from fish-killing bioassays with natural samples; growing the clones with axenic algal prey; and retesting the isolates in a second set of fish bioassays. The specific environmental conditions used (e.g., temperature, salinity, light, other factors) must remain flexible, given the wide range of conditions from which natural estuarine samples are derived. We present a comparison of information provided for fish culture conditions, reported in international science journals in which such research is routinely published, and we provide information from more than 2,000 fish bioassays with toxic Pfiesteria, along with recommendations for suitable ranges and frequency of monitoring of environmental variables. We present data demonstrating that algal assays, unlike these standardized fish bioassays, should not be used to detect toxic strains of Pfiesteria spp. Finally, we recommend how quality control/assurance can be most rapidly advanced among laboratories engaged in studies that require research-quality isolates of toxic Pfiesteria spp. PMID:11677184

Culturing conditions affect biological control activity of Trichoderma atroviride against Rhizoctonia solani in ryegrass.

PubMed

Daryaei, A; Jones, E E; Ghazalibiglar, H; Glare, T R; Falloon, R E

2016-08-01

Effects of culture conditions on productivity, germinability and bioactivity of Trichoderma atroviride LU132 conidia were assessed to identify the factors affecting conidium 'fitness' (quantity and quality) and to withstand variable environmental conditions, increase conidial productivity, and perform optimum bioactivity. The interaction effects of temperatures (20 or 30°C) vs hydrocarbon types (dextrose or sucrose in constant C : N 5 : 1) were assessed for bioactivity and colonization potential in pot experiments with ryegrass in the presence of pathogen, Rhizoctonia solani. Trichoderma atroviride produced in different culture conditions increased some growth parameters of ryegrass plant and also reduced the pathogenicity effects of R. solani. For example, Trichoderma colony produced at 20°C with sucrose increased all plant growth parameters and conidia produced at 20°C with dextrose gave the greatest bioactivity. The bimodal population cycle in T. atroviride recurred in pot experiments in a manner similar to that previously observed in agar plates but indicating that simulated natural conditions shortened the Trichoderma life cycle. Trichoderma colonized ryegrass root system and symbiotically interacted with ryegrass and greater ryegrass colonization resulted from medium production treatment with dextrose rather than sucrose. This study is the first report on the effects of inoculum production conditions on conidium quality of Trichoderma to colonize and to maintain populations in host rhizospheres, and also the ability to promote plant growth and suppress a soil-borne disease. The results of these experiments provide new knowledge on how manipulation of culture conditions of T. atroviride LU132 can influence conidium fitness, as a basis for optimizing commercial production of the fungus as a biological control agent. © 2016 The Society for Applied Microbiology.

Flow field measurements in the cell culture unit

NASA Technical Reports Server (NTRS)

Walker, Stephen; Wilder, Mike; Dimanlig, Arsenio; Jagger, Justin; Searby, Nancy

2002-01-01

The cell culture unit (CCU) is being designed to support cell growth for long-duration life science experiments on the International Space Station (ISS). The CCU is a perfused loop system that provides a fluid environment for controlled cell growth experiments within cell specimen chambers (CSCs), and is intended to accommodate diverse cell specimen types. Many of the functional requirements depend on the fluid flow field within the CSC (e.g., feeding and gas management). A design goal of the CCU is to match, within experimental limits, all environmental conditions, other than the effects of gravity on the cells, whether the hardware is in microgravity ( micro g), normal Earth gravity, or up to 2g on the ISS centrifuge. In order to achieve this goal, two steps are being taken. The first step is to characterize the environmental conditions of current 1g cell biology experiments being performed in laboratories using ground-based hardware. The second step is to ensure that the design of the CCU allows the fluid flow conditions found in 1g to be replicated from microgravity up to 2g. The techniques that are being used to take these steps include flow visualization, particle image velocimetry (PIV), and computational fluid dynamics (CFD). Flow visualization using the injection of dye has been used to gain a global perspective of the characteristics of the CSC flow field. To characterize laboratory cell culture conditions, PIV is being used to determine the flow field parameters of cell suspension cultures grown in Erlenmeyer flasks on orbital shakers. These measured parameters will be compared to PIV measurements in the CSCs to ensure that the flow field that cells encounter in CSCs is within the bounds determined for typical laboratory experiments. Using CFD, a detailed simulation is being developed to predict the flow field within the CSC for a wide variety of flow conditions, including microgravity environments. Results from all these measurements and analyses of the

Miniaturized Mass-Spectrometry-Based Analysis System for Fully Automated Examination of Conditioned Cell Culture Media

PubMed Central

Weber, Emanuel; Pinkse, Martijn W. H.; Bener-Aksam, Eda; Vellekoop, Michael J.; Verhaert, Peter D. E. M.

2012-01-01

We present a fully automated setup for performing in-line mass spectrometry (MS) analysis of conditioned media in cell cultures, in particular focusing on the peptides therein. The goal is to assess peptides secreted by cells in different culture conditions. The developed system is compatible with MS as analytical technique, as this is one of the most powerful analysis methods for peptide detection and identification. Proof of concept was achieved using the well-known mating-factor signaling in baker's yeast, Saccharomyces cerevisiae. Our concept system holds 1 mL of cell culture medium and allows maintaining a yeast culture for, at least, 40 hours with continuous supernatant extraction (and medium replenishing). The device's small dimensions result in reduced costs for reagents and open perspectives towards full integration on-chip. Experimental data that can be obtained are time-resolved peptide profiles in a yeast culture, including information about the appearance of mating-factor-related peptides. We emphasize that the system operates without any manual intervention or pipetting steps, which allows for an improved overall sensitivity compared to non-automated alternatives. MS data confirmed previously reported aspects of the physiology of the yeast-mating process. Moreover, matingfactor breakdown products (as well as evidence for a potentially responsible protease) were found. PMID:23091722

Laboratory Evaluation and Application of Microwave Absorption Properties Under Simulated Conditions for Planetary Atmospheres

NASA Technical Reports Server (NTRS)

Steffes, Paul G.

1997-01-01

Radio absorptivity data for planetary atmospheres obtained from spacecraft radio occultation experiments and earth-based radio astronomical observations can be used to infer abundances of microwave absorbing constituents in those atmospheres, as long as reliable information regarding the microwave absorbing properties of potential constituents is available. The use of theoretically-derived microwave absorption properties for such atmospheric constituents, or using laboratory measurements of such properties under environmental conditions which are significantly different than those of the planetary atmosphere being studied, often leads to significant misinterpretation of available opacity data. Laboratory measurements completed under this grant (NAGW-533), have shown that the opacity from, SO2 under simulated Venus conditions is best described by a different lineshape than was previously used in theoretical predictions. The recognition of the need to make such laboratory measurements of simulated planetary atmospheres over a range of temperatures and pressures which correspond to the altitudes probed by both radio occultation experiments and radio astronomical observations, and over a range of frequencies which correspond to those used in both radio occultation experiments and radio astronomical observations, has led to the development of a facility at Georgia Tech which is capable of making such measurements. It has been the goal of this investigation to conduct such measurements and to apply the results to a wide range of planetary observations, both spacecraft and earth-based, in order to determine the identity and abundance profiles of constituents in those planetary atmospheres.

Laboratory simulations of acid-sulfate weathering under volcanic hydrothermal conditions: Implications for early Mars.

PubMed

Marcucci, Emma C; Hynek, Brian M

2014-03-01

We have completed laboratory experiments and thermochemical equilibrium models to investigate secondary mineral formation under conditions akin to volcanic, hydrothermal acid-sulfate weathering systems. Our research used the basaltic mineralogy at Cerro Negro Volcano, Nicaragua, characterized by plagioclase, pyroxene, olivine, and volcanic glass. These individual minerals and whole-rock field samples were reacted in the laboratory with 1 molal sulfuric acid at varying temperatures (65, 150, and 200°C), fluid:rock weight ratios (1:1, 4:1, and 10:1), and durations (1-60 days). Thermochemical equilibrium models were developed using Geochemist's Workbench. To understand the reaction products and fluids, we employed scanning electron microscopy/energy dispersive spectroscopy, X-ray diffraction, and inductively coupled plasma-atomic emission spectroscopy. The results of our experiments and models yielded major alteration minerals that include anhydrite, natroalunite, minor iron oxide, and amorphous Al-Si gel. We found that variations in experimental parameters did not drastically change the suite of minerals produced; instead, abundance, size, and crystallographic shape changed. Our results also suggest that it is essential to separate phases formed during experiments from those formed during fluid evaporation to fully understand the reaction processes. Our laboratory reacted and model predicted products are consistent with the mineralogy observed at places on Mars. However, our results indicate that determination of the formation conditions requires microscopic imagery and regional context, as well as a thorough understanding of contributions from both experiment precipitation and fluid evaporation minerals.

Laboratory simulations of acid-sulfate weathering under volcanic hydrothermal conditions: Implications for early Mars

PubMed Central

Marcucci, Emma C; Hynek, Brian M

2014-01-01

We have completed laboratory experiments and thermochemical equilibrium models to investigate secondary mineral formation under conditions akin to volcanic, hydrothermal acid-sulfate weathering systems. Our research used the basaltic mineralogy at Cerro Negro Volcano, Nicaragua, characterized by plagioclase, pyroxene, olivine, and volcanic glass. These individual minerals and whole-rock field samples were reacted in the laboratory with 1 molal sulfuric acid at varying temperatures (65, 150, and 200°C), fluid:rock weight ratios (1:1, 4:1, and 10:1), and durations (1–60 days). Thermochemical equilibrium models were developed using Geochemist's Workbench. To understand the reaction products and fluids, we employed scanning electron microscopy/energy dispersive spectroscopy, X-ray diffraction, and inductively coupled plasma-atomic emission spectroscopy. The results of our experiments and models yielded major alteration minerals that include anhydrite, natroalunite, minor iron oxide, and amorphous Al-Si gel. We found that variations in experimental parameters did not drastically change the suite of minerals produced; instead, abundance, size, and crystallographic shape changed. Our results also suggest that it is essential to separate phases formed during experiments from those formed during fluid evaporation to fully understand the reaction processes. Our laboratory reacted and model predicted products are consistent with the mineralogy observed at places on Mars. However, our results indicate that determination of the formation conditions requires microscopic imagery and regional context, as well as a thorough understanding of contributions from both experiment precipitation and fluid evaporation minerals. PMID:26213665

In vitro culture of individual mouse preimplantation embryos: the role of embryo density, microwells, oxygen, timing and conditioned media.

PubMed

Kelley, Rebecca L; Gardner, David K

2017-05-01

Single embryo culture is suboptimal compared with group culture, but necessary for embryo monitoring, and culture systems should be improved for single embryos. Pronucleate mouse embryos were used to assess the effect of culture conditions on single embryo development. Single culture either before or after compaction reduced cell numbers (112.2 ± 3.1; 110.2 ± 3.5) compared with group culture throughout (127.0 ± 3.4; P < 0.05). Reduction of media volume from 20 µl to 2 µl increased blastocyst cell numbers in single embryos cultured in 5% oxygen (84.4 ± 3.2 versus 97.8 ± 2.8; P < 0.05), but not in 20% oxygen (55.2 ± 2.9 versus 57.1 ± 2.8). Culture in microwell plates for the EmbryoScope and Primo Vision time-lapse systems changed cleavage timings and increased inner cell mass cell number (24.1 ± 1.0; 23.4 ± 1.2) compared with a 2 µl microdrop (18.4 ± 1.0; P < 0.05). Addition of embryo-conditioned media to single embryos increased hatching rate and blastocyst cell number (91.5 ± 4.7 versus 113.1 ± 4.4; P < 0.01). Single culture before or after compaction is therefore detrimental; oxygen, media volume and microwells influence single embryo development; and embryo-conditioned media may substitute for group culture. Copyright © 2017 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

[Traditional and modern approaches to culture of preimplantation mammalian embryos in vitro].

PubMed

Brusentsev, E Iu; Igonina, T N; Amstislavskiĭ, S Ia

2014-01-01

This review covers the basic principles and methods of in vitro culture of preimplantation mammalian embryos. The features of in vitro development of embryos of various species of animals with allowance for the composition of nutrient media are described, with special attention paid to those species that have traditionally been consideredas laboratory (i.e., mice, rats, and hamsters). The effects of suboptimal culturing conditions of preimplantation embryos on the formation of the phenotype of individuals developed from these embryos are discussed. New approaches to optimize the conditions of the development of preimplantation mammalian embryos in vitro are analyzed.

Idaho National Laboratory Cultural Resource Monitoring Report for 2013

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

Williams, Julie B.

2013-10-01

This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during 2013. Throughout the year, thirty-eight cultural resource localities were revisited including: two locations with Native American human remains, one of which is also a cave; fourteen additional caves; seven prehistoric archaeological sites ; four historic archaeological sites; one historic trail; one nuclear resource (Experimental Breeder Reactor-I, a designated National Historic Landmark); and nine historic structures located at the Central Facilities Area. Of the monitored resources, thirty-three were routinely monitored, and five were monitored to assess project compliance with cultural resourcemore » recommendations along with the effects of ongoing project activities. On six occasions, ground disturbing activities within the boundaries of the Power Burst Facility/Critical Infrastructure Test Range Complex (PBF/CITRC) were observed by INL CRM staff prepared to respond to any additional finds of Native American human remains. In addition, two resources were visited more than once as part of the routine monitoring schedule or to monitor for additional damage. Throughout the year, most of the cultural resources monitored had no visual adverse changes resulting in Type 1determinations. However, Type 2 impacts were noted at eight sites, indicating that although impacts were noted or that a project was operating outside of culturally cleared limitations, cultural resources retained integrity and noted impacts did not threaten National Register eligibility. No new Type 3 or any Type 4 impacts that adversely impacted cultural resources and threatened National Register eligibility were observed at cultural resources monitored in 2013.« less

Chemostat Culture for Yeast Physiology.

PubMed

Kerr, Emily O; Dunham, Maitreya J

2017-07-05

The use of chemostat culture facilitates the careful comparison of different yeast strains growing in well-defined conditions. Variations in physiology can be measured by examining gene expression, metabolite levels, protein content, and cell morphology. In this protocol, we show how a combination of sample types can be collected during harvest from a single 20-mL chemostat in a ministat array, with special attention to coordinating the handling of the most time-sensitive sample types. © 2017 Cold Spring Harbor Laboratory Press.

Directing Adipose-Derived Stem Cells into Keratinocyte-Like Cells: Impact of Medium Composition and Culture Condition.

PubMed

Petry, L; Kippenberger, S; Meissner, M; Kleemann, J; Kaufmann, R; Rieger, U M; Wellenbrock, S; Reichenbach, G; Zöller, N; Valesky, E

2018-04-28

Adipose-derived stem cells (ASC) are known to transdifferentiate into a wide range of different cell species in vitro including along the epidermal lineage. This property makes them a promising tool for regenerative medicine in order to restore the epidermal barrier. The present study is dedicated to identify in vitro conditions enabling transdifferentiation to a keratinocyte-like phenotype. Especially, the impact of different culture conditions (media compositions, 2D-, 3D-cultures) and extracellular matrix (ECM) molecules was evaluated. ASC derived from subcutaneous abdominal fat were characterized by stemness associated markers and subjected to different media. Epithelial differentiation in 2D cultures was monitored by pan-cytokeratin expression using flow cytometry and immunocytochemistry. In order to evaluate the impact of different ECM molecules on epidermal stratification, 3D cultures were produced, lifted to the air-liquid-interface (ALI) and examined by histological analysis and quantitative real-time RT-PCR. We identified a medium composition containing retinoic acid, hydrocortisone, ascorbic acid and BMP-4 enabling maximum pan-cytokeratin expression in 2D cultures. Moreover, adhesion to type IV collagen further promotes the pan-cytokeratin expression. When cultures were lifted to the ALI, significant stratification was observed, particularly in supports coated with type IV collagen or fibronectin. Moreover, epidermal differentiation markers (involucrin, cytokeratin 1 and 14) become induced. Conditions with hampered wound healing such as non-healing ulcers demand new treatment regimes. The here introduced optimized protocols for transdifferentiation of ASC into keratinocyte-like cells may help to establish more effective treatment procedures. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Coupled numerical modeling of gas hydrates bearing sediments from laboratory to field-scale conditions

NASA Astrophysics Data System (ADS)

Sanchez, M. J.; Santamarina, C.; Gai, X., Sr.; Teymouri, M., Sr.

2017-12-01

Stability and behavior of Hydrate Bearing Sediments (HBS) are characterized by the metastable character of the gas hydrate structure which strongly depends on thermo-hydro-chemo-mechanical (THCM) actions. Hydrate formation, dissociation and methane production from hydrate bearing sediments are coupled THCM processes that involve, amongst other, exothermic formation and endothermic dissociation of hydrate and ice phases, mixed fluid flow and large changes in fluid pressure. The analysis of available data from past field and laboratory experiments, and the optimization of future field production studies require a formal and robust numerical framework able to capture the very complex behavior of this type of soil. A comprehensive fully coupled THCM formulation has been developed and implemented into a finite element code to tackle problems involving gas hydrates sediments. Special attention is paid to the geomechanical behavior of HBS, and particularly to their response upon hydrate dissociation under loading. The numerical framework has been validated against recent experiments conducted under controlled conditions in the laboratory that challenge the proposed approach and highlight the complex interaction among THCM processes in HBS. The performance of the models in these case studies is highly satisfactory. Finally, the numerical code is applied to analyze the behavior of gas hydrate soils under field-scale conditions exploring different features of material behavior under possible reservoir conditions.

Biological responses of the marine diatom Chaetoceros socialis to changing environmental conditions: A laboratory experiment

PubMed Central

Roevros, Nathalie; Dehairs, Frank; Chou, Lei

2017-01-01

Diatoms constitute a major group of phytoplankton, accounting for ~20% of the world’s primary production. It has been shown that iron (Fe) can be the limiting factor for phytoplankton growth, in particular, in the HNLC (High Nutrient Low Chlorophyll) regions. Iron plays thus an essential role in governing the marine primary productivity and the efficiency of biological carbon pump. Oceanic systems are undergoing continuous modifications at varying rates and magnitudes as a result of changing climate. The objective of our research is to evaluate how changing environmental conditions (dust deposition, ocean warming and acidification) can affect marine Fe biogeochemistry and diatom growth. Laboratory culture experiments using a marine diatom Chaetoceros socialis were conducted at two temperatures (13°C and 18°C) and under two pCO2 (carbon dioxide partial pressure) (400 μatm and 800 μatm) conditions. The present study clearly highlights the effect of ocean acidification on enhancing the release of Fe upon dust deposition. Our results also confirm that being a potential source of Fe, dust provides in addition a readily utilizable source of macronutrients such as dissolved phosphate (PO4) and silicate (DSi). However, elevated atmospheric CO2 concentrations may also have an adverse impact on diatom growth, causing a decrease in cell size and possible further changes in phytoplankton composition. Meanwhile, ocean warming may lead to the reduction of diatom production and cell size, inducing poleward shifts in the biogeographic distribution of diatoms. The changing climate has thus a significant implication for ocean phytoplankton growth, cell size and primary productivity, phytoplankton distribution and community composition, and carbon (C), nitrogen (N), phosphorus (P), silicon (Si) and Fe biogeochemical cycles in various ways. PMID:29190826

Dynamic cell culture system (7-IML-1)

NASA Technical Reports Server (NTRS)

Cogoli, Augusto

1992-01-01

This experiment is one of the Biorack experiments being flown on the International Microgravity Laboratory 1 (MIL-1) mission as part of an investigation studying cell proliferation and performance in space. One of the objectives of this investigation is to assess the potential benefits of bioprocessing in space with the ultimate goal of developing a bioreactor for continuous cell cultures in space. This experiment will test the operation of an automated culture chamber that was designed for use in a Bioreactor in space. The device to be tested is called the Dynamic Cell Culture System (DCCS). It is a simple device in which media are renewed or chemicals are injected automatically, by means of osmotic pumps. This experiment uses four Type I/O experiment containers. One DCCS unit, which contains a culture chamber with renewal of medium and a second chamber without a medium supply fits in each container. Two DCCS units are maintained under zero gravity conditions during the on-orbit period. The other two units are maintained under 1 gh conditions in a 1 g centrifuge. The schedule for incubator transfer is given.

Effects of Lactobacillus plantarum Strain OLL2712 Culture Conditions on the Anti-inflammatory Activities for Murine Immune Cells and Obese and Type 2 Diabetic Mice.

PubMed

Toshimitsu, T; Ozaki, S; Mochizuki, J; Furuichi, K; Asami, Y

2017-04-01

Studies on the health-promoting effects of lactic acid bacteria (LAB) are numerous, but few provide examples of the relationship between LAB function and culture conditions. We verified the effect of differences in culture conditions on Lactobacillus plantarum OLL2712 functionality; this strain exhibits anti-inflammatory activity and preventive effects against metabolic disorders. We measured interleukin-10 (IL-10) and IL-12 production in murine immune cells treated with OLL2712 cells prepared under various culture conditions. The results showed that the IL-10-inducing activities of OLL2712 cells on murine immune cells differed dramatically between OLL2712 groups at different culture phases and using different culture medium components, temperatures, and neutralizing pHs. In particular, exponential-phase cells had much more IL-10-inducing activity than stationary-phase cells. We confirmed that the Toll-like receptor 2 (TLR2) stimulation activity of OLL2712 cells depended on culture conditions in conjunction with IL-10-inducing activity. We also demonstrated functional differences by culture phases in vivo ; OLL2712 cells at exponential phase had more anti-inflammatory activity and anti-metabolic-disorder effects on obese and diabetic mice than those by their stationary-phase counterparts. These results suggest that culture conditions affect the functionality of anti-inflammatory LAB. IMPORTANCE While previous studies demonstrated that culture conditions affected the immunomodulatory properties of lactic acid bacteria (LAB), few have comprehensively investigated the relationship between culture conditions and LAB functionality. In this study, we demonstrated several culture conditions of Lactobacillus plantarum OLL2712 for higher anti-inflammatory activity. We also showed that culture conditions concretely influenced the health-promoting functions of OLL2712 in vivo , particularly against metabolic disorders. Further, we characterized a novel mechanism by which

Ligninolytic enzyme production in selected sub-tropical white rot fungi under different culture conditions.

PubMed

Tekere, M; Zvauya, R; Read, J S

2001-01-01

Lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase activities in selected sub-tropical white rot fungal species from Zimbabwe were determined. The enzyme activities were assayed at varying concentrations of C, N and Mn2+. Manganese peroxidase and laccase activities were the only expressed activities in the fungi under the culture conditions tested. Trametes species, T. cingulata, T. elegans and T. pocas produced the highest manganese peroxidase activities in a medium containing high carbon and low nitrogen conditions. High nitrogen conditions favoured high manganese peroxidase activity in DSPM95, L. velutinus and Irpex spp. High manganese peroxidase activity was notable for T. versicolor when both carbon and nitrogen in the medium were present at high levels. Laccase production by the isolates was highest under conditions of high nitrogen and those conditions with both nitrogen and carbon at high concentration. Mn2+ concentrations between 11-25 ppm gave the highest manganese peroxidase activity compared to a concentration of 40 ppm or when there was no Mn2+ added. Laccase activity was less influenced by Mn2+ levels. While some laccase activity was produced in the absence of Mn2+, the enzyme levels were higher when Mn2+ was added to the culture medium.

Primary Human Uterine Leiomyoma Cell Culture Quality Control: Some Properties of Myometrial Cells Cultured under Serum Deprivation Conditions in the Presence of Ovarian Steroids.

PubMed

Bonazza, Camila; Andrade, Sheila Siqueira; Sumikawa, Joana Tomomi; Batista, Fabrício Pereira; Paredes-Gamero, Edgar J; Girão, Manoel J B C; Oliva, Maria Luiza V; Castro, Rodrigo Aquino

2016-01-01

Cell culture is considered the standard media used in research to emulate the in vivo cell environment. Crucial in vivo experiments cannot be conducted in humans and depend on in vitro methodologies such as cell culture systems. However, some procedures involving the quality control of cells in culture have been gradually neglected by failing to acknowledge that primary cells and cell lines change over time in culture. Thus, we report methods based on our experience for monitoring primary cell culture of human myometrial cells derived from uterine leiomyoma. We standardized the best procedure of tissue dissociation required for the study of multiple genetic marker systems that include species-specific antigens, expression of myofibroblast or myoblast markers, growth curve, serum deprivation, starvation by cell cycle synchronization, culture on collagen coated plates, and 17 β-estradiol (E2) and progesterone (P4) effects. The results showed that primary myometrial cells from patients with uterine leiomyoma displayed myoblast phenotypes before and after in vitro cultivation, and leiomyoma cells differentiated into mature myocyte cells under the appropriate differentiation-inducing conditions (serum deprivation). These cells grew well on collagen coated plates and responded to E2 and P4, which may drive myometrial and leiomyoma cells to proliferate and adhere into a focal adhesion complex involvement in a paracrine manner. The establishment of these techniques as routine procedures will improve the understanding of the myometrial physiology and pathogenesis of myometrium-derived diseases such as leiomyoma. Mimicking the in vivo environment of fibrotic conditions can prevent false results and enhance results that are based on cell culture integrity.

Increased Risk of Genetic and Epigenetic Instability in Human Embryonic Stem Cells Associated with Specific Culture Conditions

PubMed Central

Garitaonandia, Ibon; Amir, Hadar; Boscolo, Francesca Sesillo; Wambua, Gerald K.; Schultheisz, Heather L.; Sabatini, Karen; Morey, Robert; Waltz, Shannon; Wang, Yu-Chieh; Tran, Ha; Leonardo, Trevor R.; Nazor, Kristopher; Slavin, Ileana; Lynch, Candace; Li, Yingchun; Coleman, Ronald; Gallego Romero, Irene; Altun, Gulsah; Reynolds, David; Dalton, Stephen; Parast, Mana; Loring, Jeanne F.; Laurent, Louise C.

2015-01-01

The self-renewal and differentiation capacities of human pluripotent stem cells (hPSCs) make them a promising source of material for cell transplantation therapy, drug development, and studies of cellular differentiation and development. However, the large numbers of cells necessary for many of these applications require extensive expansion of hPSC cultures, a process that has been associated with genetic and epigenetic alterations. We have performed a combinatorial study on both hESCs and hiPSCs to compare the effects of enzymatic vs. mechanical passaging, and feeder-free vs. mouse embryonic fibroblast feeder substrate, on the genetic and epigenetic stability and the phenotypic characteristics of hPSCs. In extensive experiments involving over 100 continuous passages, we observed that both enzymatic passaging and feeder-free culture were associated with genetic instability, higher rates of cell proliferation, and persistence of OCT4/POU5F1-positive cells in teratomas, with enzymatic passaging having the stronger effect. In all combinations of culture conditions except for mechanical passaging on feeder layers, we noted recurrent deletions in the genomic region containing the tumor suppressor gene TP53, which was associated with decreased mRNA expression of TP53, as well as alterations in the expression of several downstream genes consistent with a decrease in the activity of the TP53 pathway. Among the hESC cultures, we also observed culture-associated variations in global gene expression and DNA methylation. The effects of enzymatic passaging and feeder-free conditions were also observed in hiPSC cultures. Our results highlight the need for careful assessment of the effects of culture conditions on cells intended for clinical therapies. PMID:25714340

Methods and Compositions Based on Culturing Microorganisms in Low Sedimental Fluid Shear Conditions

NASA Technical Reports Server (NTRS)

Ott, C. Mark; Nickerson, Cheryl A.; Wilson, James W.; Sarker, Shameema; Nauman, Eric A.; Schurr, Michael J.; Nelman-Gonzalez, Mayra A.

2012-01-01

The benefits of applying a low sedimental fluid shear environment to manipulate microorganisms were examined. Microorganisms obtained from a low sedimental fluid shear culture, which exhibit modified phenotypic and molecular genetic characteristics, are useful for the development of novel and improved diagnostics, therapeutics, vaccines, and bio-industrial products. Furthermore, application of low sedimental fluid conditions to microorganisms permits identification of molecules uniquely expressed under these conditions, providing a basis for the design of new therapeutic targets.

Laboratory evaluation and application of microwave absorption properties under simulated conditions for planetary atmospheres

NASA Technical Reports Server (NTRS)

Steffes, P. G.

1986-01-01

The recognition of the need to make laboratory measurements of simulated planetary atmospheres over a range of temperatures and pressure which correspond to the altitudes probed by radio occultation experiments, and over a range of frequencies which correspond to both radio occultation experiments and radio astronomical observations, has led to the development of a facility at Georgia Tech which is capable of making such measurements. Construction was completed of the outer planets simulator and measurements were conducted of the microwave absorption and refraction from nitrogen under simulated Titan conditions. The results of these and previous laboratory measurements were applied to a wide range of microwave opacity measurements, in order to derive constituent densities and distributions in planetary atmospheres such as Venus.

Differential Radiosensitizing Effect of Valproic Acid in Differentiation Versus Self-Renewal Promoting Culture Conditions

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

Debeb, Bisrat G.; Xu Wei; Mok, Henry

2010-03-01

Purpose: It has been shown that valproic acid (VA) enhances the proliferation and self-renewal of normal hematopoietic stem cells and that breast cancer stem/progenitor cells can be resistant to radiation. From these data, we hypothesized that VA would fail to radiosensitize breast cancer stem/progenitor cells grown to three-dimensional (3D) mammospheres. Methods and Materials: We used the MCF7 breast cancer cell line grown under stem cell-promoting culture conditions (3D mammosphere) and standard nonstem cell monolayer culture conditions (two-dimensional) to examine the effect of pretreatment with VA on radiation sensitivity in clonogenic survival assays and on the expression of embryonic stem cellmore » transcription factors. Results: 3D-cultured MCF-7 cells expressed higher levels of Oct4, Nanog, and Sox2. The 3D passage enriched self-renewal and increased radioresistance in the 3D mammosphere formation assays. VA radiosensitized adherent cells but radioprotected 3D cells in single-fraction clonogenic assays. Moreover, fractionated radiation sensitized VA-treated adherent MCF7 cells but did not have a significant effect on VA-treated single cells grown to mammospheres. Conclusion: We have concluded that VA might preferentially radiosensitize differentiated cells compared with those expressing stem cell surrogates and that stem cell-promoting culture is a useful tool for in vitro evaluation of novel cancer therapeutic agents and radiosensitizers.« less

Bronchoscopic culture

MedlinePlus

... a laboratory exam to check a piece of tissue or fluid from the lungs for infection-causing germs. ... Culture - bronchoscopic ... used to get a sample ( biopsy ) of lung tissue or fluid. The sample ... a special dish (culture). It is then watched to see if bacteria ...

Effect of Culture Conditions on Viability of Mouse and Rat Embryos Developed in Vitro

PubMed Central

Popova, Elena; Bader, Michael; Krivokharchenko, Alexander

2011-01-01

Currently in vitro culture of mouse preimplantation embryos has become a very important technique to investigate different mechanisms of early embryogenesis. However, there is a big difference in the preimplantation development between mammalian species. Despite close relatedness to mice, in vitro cultivation of rat preimplantation embryos is still delicate and needs further investigation and optimizations. In this study we have compared the in vitro developmental potential of mouse and rat embryos cultured at different culture conditions in parallel experiments. Interestingly, mouse zygotes developed in vitro until blastocyst stage even in inadequate medium without any phosphates and with low osmolarity which was formulated especially for cultivation of rat embryos. Rat parthenotes and zygotes developed in M16 medium formulated for mouse embryos only till 2-cell stage and further development is blocked completely at this stage. Moreover, developmental ability of rat embryos in vitro was significantly lower in comparison with mouse even in special rat mR1ECM medium. Mouse and rat embryos at 2-cell stage obtained in vivo developed until blastocyst stages significantly more efficiently compared to zygotes. Culture of mouse zygotes in glass capillaries resulted in a significantly higher rate of morula and blastocyst development compared with dishes. The Well-of-the-Well system resulted in a significant improvement when compared with dishes for the culture of rat zygotes only until morula stage. Reduced oxygen tension increased the developmental rate of rat but not mouse zygotes until blastocyst stage. This study demonstrates that development of early preimplantation embryos is altered by different culture conditions and show strong differences even between two related species such as mice and rats. Therefore, for understanding the fundamental mechanisms of early mammalian development it is very important to use embryos of various species. PMID:24710194

Cardiac Cells Beating in Culture: A Laboratory Exercise

ERIC Educational Resources Information Center

Weaver, Debora

2007-01-01

This article describes how to establish a primary tissue culture, where cells are taken directly from an organ of a living animal. Cardiac cells are taken from chick embryos and transferred to culture dishes. These cells are not transformed and therefore have a limited life span. However, the unique characteristics of cardiac cells are maintained…

Experimental and theoretical study of microalgal competition in laboratory and natural ecosystems

NASA Astrophysics Data System (ADS)

Pisman, T. I.; Somova, L. A.

An important aspect of studying mixed cultures of microalgae is the artificial ecosystems containing algal culture as a regeneration link and a source of vegetable substances. The peculiarities of studying the stability of microalgae mixed cultures in the laboratory and natural environment have been considered in the work. The role of factors most essentially affecting the species structure of phytoplankton community (temperature factor, light intensity, pH environments, elements of mineral nutrition, algal metabolites, predation and fluctuation of environmental conditions) has been displayed. As a result of experimental and theoretical modelling of the microalgae Chlorella vulgaris and Scenedesmus quadricauda competition under limitation on nitrogen, the impossibility of their co-existence has been revealed. Under these conditions Chl. vulgaris turned out to be less competitive than Sc. quadricauda. The influence of the ratio of biogenic elements concentration in the environment, which should be recognized as an independent regulatory factor limiting growth of populations in the community and, thus affecting its structure, has been analyzed.

Culture - joint fluid

MedlinePlus

... this page: //medlineplus.gov/ency/article/003742.htm Culture - joint fluid To use the sharing features on this page, please enable JavaScript. Joint fluid culture is a laboratory test to detect infection-causing ...

Morphometric changes of Triatoma flavida Neiva, 1911 (Hemiptera:Triatominae) in the transition from sylvatic to laboratory conditions.

PubMed

Rodríguez Rodríguez, Jinnay; Fuentes González, Omar; Nodarse, Jorge Fraga; Monzote Fidalgo, Lianet; Dujardin, Jean-Pierre

2007-01-01

The one-generational metric changes occurring in Triatoma flavida (Hemiptera: Triatominae) when carried from its wild habitat (caves) to laboratory, were examined using traditional morphometric techniques. As for other species of Triatoma, Rhodnius or Panstrongylus studied in similar conditions, a significant reduction of head, thorax and wing size was observed. Sexual dimorphism of the wings, while present in the wild sample, was not detected anymore in the laboratory individuals. Biological significance and epidemiological importance are discussed.

Developing best practice for fungal specimen management: audit of UK microbiology laboratories.

PubMed

Lasseter, G; Palmer, M; Morgan, J; Watts, J; Yoxall, H; Kibbler, C; McNulty, C

2011-01-01

This study represents an audit of microbiology laboratories in the UK to ascertain whether they are aware of, or follow, the Health Protection Agency (HPA) National Standard Methods Standard Operating Procedure (NSM SOP) for the investigation of dermatological specimens for superficial mycoses, or use a locally adapted version. A questionnaire audit was distributed to 179 NHS microbiology laboratories throughout England, Wales, Scotland and Northern Ireland. The NSM SOP was followed by 92% of laboratories for the microscopy of dermatological samples; light microscopy/ KOH digestion was used by 63% and fluorescence microscopy/KOH digestion by 29% of laboratories. Preliminary reports post-microscopy were issued by 98% of laboratories, with 93% issuing reports within 48 hours. Adherence to the NSM SOP guidelines for culture was low; only 34% of laboratories incubated microscopy-negative specimens for the recommended 14 days, while approximately 60% incubated microscopy-positive specimens for 21 days. The culture medium recommended by the NSM SOP was used in 82% of laboratories. Comments were added to culture reports by 51% of laboratories; most were added manually and comments varied between laboratories. Nail samples were the most common sample received from primary care, followed by skin and hair. These results show no significant difference in the rate of microscopy positives versus culture positives. Microscopy and culture are the easiest and cheapest methods available to UK laboratories for the investigation of suspected superficial fungal infections. Although most laboratories included in this audit claimed to follow the NSM SOP for microscopy and culture, these results show that the techniques used vary throughout the UK. To maximise the service provided to primary care, UK laboratories should use standardise methods based on the NSM SOP.

Quality evaluation of green tea leaf cultured under artificial light condition using gas chromatography/mass spectrometry.

PubMed

Miyauchi, Shunsuke; Yonetani, Tsutomu; Yuki, Takayuki; Tomio, Ayako; Bamba, Takeshi; Fukusaki, Eiichiro

2017-02-01

For an experimental model to elucidate the relationship between light quality during plant culture conditions and plant quality of crops or vegetables, we cultured tea plants (Camellia sinensis) and analyzed their leaves as tea material. First, metabolic profiling of teas from a tea contest in Japan was performed with gas chromatography/mass spectrometry (GC/MS), and then a ranking predictive model was made which predicted tea rankings from their metabolite profile. Additionally, the importance of some compounds (glutamine, glutamic acid, oxalic acid, epigallocatechin, phosphoric acid, and inositol) was elucidated for measurement of the quality of tea leaf. Subsequently, tea plants were cultured in artificial conditions to control these compounds. From the result of prediction by the ranking predictive model, the tea sample supplemented with ultraviolet-A (315-399 nm) showed the highest ranking. The improvement in quality was thought to come from the high amino-acid and decreased epigallocatechin content in tea leaves. The current study shows the use and value of metabolic profiling in the field of high-quality crops and vegetables production that has been conventionally evaluated by human sensory analysis. Metabolic profiling enables us to form hypothesis to understand and develop high quality plant cultured under artificial condition. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Bacillus Classification Based on Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry-Effects of Culture Conditions.

PubMed

Shu, Lin-Jie; Yang, Yu-Liang

2017-11-14

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a reliable and rapid technique applied widely in the identification and classification of microbes. MALDI-TOF MS has been used to identify many endospore-forming Bacillus species; however, endospores affect the identification accuracy when using MALDI-TOF MS because they change the protein composition of samples. Since culture conditions directly influence endospore formation and Bacillus growth, in this study we clarified how culture conditions influence the classification of Bacillus species by using MALDI-TOF MS. We analyzed members of the Bacillus subtilis group and Bacillus cereus group using different incubation periods, temperatures and media. Incubation period was found to affect mass spectra due to endospores which were observed mixing with vegetative cells after 24 hours. Culture temperature also resulted in different mass spectra profiles depending on the temperature best suited growth and sporulation. Conversely, the four common media for Bacillus incubation, Luria-Bertani agar, nutrient agar, plate count agar and brain-heart infusion agar did not result in any significant differences in mass spectra profiles. Profiles in the range m/z 1000-3000 were found to provide additional data to the standard ribosomal peptide/protein region m/z 3000-15000 profiles to enable easier differentiation of some highly similar species and the identification of new strains under fresh culture conditions. In summary, control of culture conditions is vital for Bacillus identification and classification by MALDI-TOF MS.

Primary Human Uterine Leiomyoma Cell Culture Quality Control: Some Properties of Myometrial Cells Cultured under Serum Deprivation Conditions in the Presence of Ovarian Steroids

PubMed Central

Sumikawa, Joana Tomomi; Batista, Fabrício Pereira; Paredes-Gamero, Edgar J.; Girão, Manoel J. B. C.; Oliva, Maria Luiza V.

2016-01-01

Cell culture is considered the standard media used in research to emulate the in vivo cell environment. Crucial in vivo experiments cannot be conducted in humans and depend on in vitro methodologies such as cell culture systems. However, some procedures involving the quality control of cells in culture have been gradually neglected by failing to acknowledge that primary cells and cell lines change over time in culture. Thus, we report methods based on our experience for monitoring primary cell culture of human myometrial cells derived from uterine leiomyoma. We standardized the best procedure of tissue dissociation required for the study of multiple genetic marker systems that include species-specific antigens, expression of myofibroblast or myoblast markers, growth curve, serum deprivation, starvation by cell cycle synchronization, culture on collagen coated plates, and 17 β-estradiol (E2) and progesterone (P4) effects. The results showed that primary myometrial cells from patients with uterine leiomyoma displayed myoblast phenotypes before and after in vitro cultivation, and leiomyoma cells differentiated into mature myocyte cells under the appropriate differentiation-inducing conditions (serum deprivation). These cells grew well on collagen coated plates and responded to E2 and P4, which may drive myometrial and leiomyoma cells to proliferate and adhere into a focal adhesion complex involvement in a paracrine manner. The establishment of these techniques as routine procedures will improve the understanding of the myometrial physiology and pathogenesis of myometrium-derived diseases such as leiomyoma. Mimicking the in vivo environment of fibrotic conditions can prevent false results and enhance results that are based on cell culture integrity. PMID:27391384

Successful Consecutive Expansion of Limbal Explants Using a Biosafe Culture Medium under Feeder Layer-Free Conditions.

PubMed

López-Paniagua, Marina; Nieto-Miguel, Teresa; de la Mata, Ana; Galindo, Sara; Herreras, José M; Corrales, Rosa M; Calonge, Margarita

2017-05-01

Transplantation of in vitro cultured limbal epithelial stem cells (LESCs) is a treatment widely used for LESC deficiency. However, the number of limbal tissue donors is limited, and protocols for LESC cultivation often include compounds and/or feeder layers that can induce side effects and/or increase the cost of the culture procedure. We investigated the feasibility of obtaining more than one limbal primary culture (LPC) from the same biopsy using a culture medium in which several potentially harmful compounds were replaced at the same time by biosafe supplements, allowing the LESC cultivation without feeder layers. We established feeder layer-free LPCs with three culture media: (1) a modified supplemental hormonal epithelial medium, containing potential harmful components (cholera toxin, dimethylsulfoxide, and fetal bovine serum [FBS]), (2) IOBA-FBS, a medium with FBS but with no other harmful supplements, and (3) IOBA-HS, similar to IOBA-FBS but with human serum instead of FBS. Additionally, the same limbal explant was consecutively cultured with IOBA-HS producing three cultures. LPCs were characterized by real-time reverse transcription polymerase chain reaction and/or immunofluorescence. LPCs cultured with the three media under feeder layer-free conditions showed cuboidal cells and no significant differences in the percentage of positive cells for limbal (ABCG2, p63, and K14) and corneal (K3, K12) proteins. Except for ABCG2, the relative mRNA expression of the LESC markers was significantly higher when IOBA-FBS or IOBA-HS was used. LPC1 showed characteristics similar to LPC0, while LPC2 cell morphology became elongated and the expression of some LESC markers was diminished. IOBA-HS enables the culturing of up to two biosafe homologous LPCs from one limbal tissue under feeder layer-free conditions. The routine use of this culture medium could improve both the biosafety and the number of available LPCs for potential clinical transplantation, as well as decrease

A study of 6S workplace improvement in Ergonomic Laboratory

NASA Astrophysics Data System (ADS)

Sari, AD; Suryoputro, MR; Rahmillah, FI

2017-12-01

This article discusses 6S implementation in Ergonomic Laboratory, Department of Industrial Engineering, Islamic University of Indonesia. This research is improvement project of 5S implementation in Ergonomic laboratory. Referring to the 5S implementation of the previous year, there have been improvements from environmental conditions or a more organized workplace however there is still a lack of safety aspects. There are several safeties problems such as equipment arrangement, potential hazards of room dividers that cause injury several times, placement of fire extinguisher, no evacuation path and assembly point in case of fire, as well as expired hydrant condition and lack of awareness of stakeholders related to safety. Therefore, this study aims to apply the 6S kaizen method to the Ergonomic laboratory to facilitate the work process, reduce waste, improve work safety and improve staff performance. Based on the score 6S assessment increased audit results by 32 points, before implementation is 75 point while after implementation is 107 point. This has implications for better use for mitigate people in laboratory area, save time when looking for tools and materials, safe workplace, as well as improving the culture and spirit of ‘6S’ on staff due to better and safetier working environment.

Luria in Uzbekistan: the vicissitudes of cross-cultural neuropsychology.

PubMed

Nell, V

1999-03-01

If the material conditions of culture shape cognitive structures, as Luria and Vygotsky argued, the "extraordinarily deep and rapid restructuring of historical forms" (Luria, 1971, 265) in the Soviet Republics that followed the Bolshevik revolution of 1917 provided a natural laboratory to determine whether processes of modernization changed traditional ways of thinking. This was the purpose of Luria's 1931 expedition to the Soviet Republic of Uzbekistan in central Asia. Luria's initial reports attracted vitriolic criticism because he had allegedly belittled "primitive" Uzbeki culture. The lasting importance of the Uzbek expedition is its emphasis on culture as a determinant of cognitive processes that remains valid to the present: in 1984, Gilbert replicated Luria's field studies in South Africa with near-identical results. Yet current neuropsychology has been slow to recognize the need for culturally sensitive assessment.

Induction of a photomixotrophic plant cell culture of Helianthus annuus and optimization of culture conditions for improved α-tocopherol production.

PubMed

Geipel, Katja; Song, Xue; Socher, Maria Lisa; Kümmritz, Sibylle; Püschel, Joachim; Bley, Thomas; Ludwig-Müller, Jutta; Steingroewer, Juliane

2014-03-01

Tocopherols, collectively known as vitamin E, are lipophilic antioxidants, which are synthesized only by photosynthetic organisms. Due to their enormous potential to protect cells from oxidative damage, tocopherols are used, e.g., as nutraceuticals and additives in pharmaceuticals. The most biologically active form of vitamin E is α-tocopherol. Most tocopherols are currently produced via chemical synthesis. Nevertheless, this always results in a racemic mixture of different and less effective stereoisomers because the natural isomer has the highest biological activity. Therefore, tocopherols synthesized in natural sources are preferred for medical purposes. The annual sunflower (Helianthus annuus L.) is a well-known source for α-tocopherol. Within the presented work, sunflower callus and suspension cultures were established growing under photomixotrophic conditions to enhance α-tocopherol yield. The most efficient callus induction was achieved with sunflower stems cultivated on solid Murashige and Skoog medium supplemented with 30 g l(-1) sucrose, 0.5 mg l(-1) of the auxin 1-naphthalene acetic acid, and 0.5 mg l(-1) of the cytokinin 6-benzylaminopurine. Photomixotrophic sunflower suspension cultures were induced by transferring previously established callus into liquid medium. The effects of light intensity, sugar concentration, and culture age on growth rate and α-tocopherol synthesis rate were characterized. A considerable increase (max. 230%) of α-tocopherol production in the cells was obtained within the photomixotrophic cell culture compared to a heterotrophic cell culture. These results will be useful for improving α-tocopherol yields of plant in vitro cultures.

Dynamically triggered slip leading to sustained fault gouge weakening under laboratory shear conditions

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

Johnson, Paul Allan

We investigate dynamic wave-triggered slip under laboratory shear conditions. The experiment is composed of a three-block system containing two gouge layers composed of glass beads and held in place by a fixed load in a biaxial configuration. When the system is sheared under steady state conditions at a normal load of 4 MPa, we find that shear failure may be instantaneously triggered by a dynamic wave, corresponding to material weakening and softening if the system is in a critical shear stress state (near failure). Following triggering, the gouge material remains in a perturbed state over multiple slip cycles as evidencedmore » by the recovery of the material strength, shear modulus, and slip recurrence time. This work suggests that faults must be critically stressed to trigger under dynamic conditions and that the recovery process following a dynamically triggered event differs from the recovery following a spontaneous event.« less

Dynamically triggered slip leading to sustained fault gouge weakening under laboratory shear conditions

DOE PAGES

Johnson, Paul Allan

2016-02-28

We investigate dynamic wave-triggered slip under laboratory shear conditions. The experiment is composed of a three-block system containing two gouge layers composed of glass beads and held in place by a fixed load in a biaxial configuration. When the system is sheared under steady state conditions at a normal load of 4 MPa, we find that shear failure may be instantaneously triggered by a dynamic wave, corresponding to material weakening and softening if the system is in a critical shear stress state (near failure). Following triggering, the gouge material remains in a perturbed state over multiple slip cycles as evidencedmore » by the recovery of the material strength, shear modulus, and slip recurrence time. This work suggests that faults must be critically stressed to trigger under dynamic conditions and that the recovery process following a dynamically triggered event differs from the recovery following a spontaneous event.« less

[Effects of culture conditions on biomass and active components of adventitious roots culture in Panax ginseng].

PubMed

Huang, Tao; Gao, Wenyuan; Wang, Juan; Cao, Yu

2010-01-01

To optimize the culture condition of adventitious roots of Panax ginseng. The adventitious roots were obtained through tissue culture by manipulation of inoculum, various sucrose concentrations and salt strength. The contents of ginsenosides Re, Rb1 and Rg1 were determined by HPLC while the contents of polysaccharides were determined by ultraviolet spectrophotometry. The multiplication of adventitious roots reached the peak when the inoculum was 20 g x L(-1). The effects of sucrose concentration and salt strength on adventitious roots were observed. The contents of polysaccharides were higher when the medium contained more sucrose. 40 g x L(-1) sucrose was favorable for roots growth and biosynthesis of Re, while 30 g x L(-1) was favorable for the biosynthesis of Rb1 and Rg1. 3/4MS medium was benefit for the growth of adventitious roots and the biosynthesis of ginsenosides. The contents of polysaccharides were decreased with the increase of salt strength. The results showed that inoculum, various sucrose concentrations and salt strength have significant influences on adventitious roots growth, secondary metabolite and polysaccharide synthesis in P. ginseng.

In Vitro Conservation of Date Palm Shoot-Tip Explants and Callus Cultures Under Minimal Growth Conditions.

PubMed

El-Dawayati, Maiada M

2017-01-01

Date palm fruit production has great economic significance for many countries. There is a fundamental necessity to conserve valuable date palm germplasm, but there are various problems with in vivo and ex situ conservation. In vitro storage has several advantages over conventional germplasm conservation methods. The in vitro technique offers a developed method of slow-growth storage, which is considered as an alternate solution for short- and medium-term storage of date palm germplasm under controlled conditions. Minimal growth conditions for germplasm conservation are generally achieved by reducing growth rate through modification of environmental growing conditions and culture, by using low temperatures, and the addition of growth retardants and osmotic agents. This chapter describes a protocol for short-term in vitro conservation of date palm shoot-tip and callus cultures under slow-growth storage conditions, using sucrose as an osmotic agent and abscisic acid (ABA) as a growth retardant at 15 °C for 12 months.

The laboratory environmental algae pond simulator (LEAPS) photobioreactor: Validation using outdoor pond cultures of Chlorella sorokiniana and Nannochloropsis salina

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

Huesemann, M.; Williams, P.; Edmundson, S.

A bench-scale photobioreactor system, termed Laboratory Environmental Algae Pond Simulator (LEAPS), was designed and constructed to simulate outdoor pond cultivation for a wide range of geographical locations and seasons. The LEAPS consists of six well-mixed glass column photobioreactors sparged with CO2-enriched air to maintain a set-point pH, illuminated from above by a programmable multicolor LED lighting (0 to 2,500 µmol/m2-sec), and submerged in a temperature controlled water-bath (-2 °C to >60 °C). Measured incident light intensities and water temperatures deviated from the respective light and temperature set-points on average only 2.3% and 0.9%, demonstrating accurate simulation of light and temperaturemore » conditions measured in outdoor ponds. In order to determine whether microalgae strains cultured in the LEAPS exhibit the same linear phase biomass productivity as in outdoor ponds, Chlorella sorokiniana and Nannochloropsis salina were cultured in the LEAPS bioreactors using light and temperature scripts measured previously in the respective outdoor pond studies. For Chlorella sorokiniana, the summer season biomass productivity in the LEAPS was 6.6% and 11.3% lower than in the respective outdoor ponds in Rimrock, Arizona, and Delhi, California; however, these differences were not statistically significant. For Nannochloropsis salina, the winter season biomass productivity in the LEAPS was statistically significantly higher (15.2%) during the 27 day experimental period than in the respective outdoor ponds in Tucson, Arizona. However, when considering only the first 14 days, the LEAPS biomass productivity was only 9.2% higher than in the outdoor ponds, a difference shown to be not statistically significant. Potential reasons for the positive or negative divergence in LEAPS performance, relative to outdoor ponds, are discussed. To demonstrate the utility of the LEAPS in predicting productivity, two other strains – Scenedesmus obliquus and

Laboratory Evaluation and Application of Microwave Absorption Properties Under Simulated Conditions for Planetary Atmospheres

NASA Technical Reports Server (NTRS)

Steffes, Paul G.

1998-01-01

Radio absorptivity data for planetary atmospheres obtained from spacecraft radio occultation experiments, entry probe radio signal absorption measurements, and earth-based radio astronomical observations can be used to infer abundances of microwave absorbing constituents in those atmospheres, as long as reliable information regarding the microwave absorbing properties of potential constituents is available. The use of theoretically-derived microwave absorption properties for such atmospheric constituents, or using laboratory measurements of such properties taken under environmental conditions which are significantly different than those of the planetary atmosphere being studied, often leads to significant misinterpretation of available opacity data. For example, laboratory measurements completed recently by Kolodner and Steffes (ICARUS 132, pp. 151-169, March 1998, attached as Appendix A) under this grant (NAGS-4190), have shown that the opacity from gaseous H2SO4 under simulated Venus conditions is best described by a different formalism than was previously used. The recognition of the need to make such laboratory measurements of simulated planetary atmospheres over a range of temperatures and pressures which correspond to the altitudes probed by both spacecraft entry probe and orbiter radio occultation experiments and by radio astronomical observations, and over a range of frequencies which correspond to those used in such experiments, has led to the development of a facility at Georgia Tech which is capable of making such measurements. It has been the goal of this investigation to conduct such measurements and to apply the results to a wide range of planetary observations, both spacecraft and earth-based, in order to determine the identity and abundance profiles of constituents in those planetary atmospheres.

A comparison of residual smear layer and erosion following different endodontic irrigation protocols tested under clinical and laboratory conditions.

PubMed

Cehreli, Zafer C; Uyanik, M Ozgur; Nagas, Emre; Tuncel, Behram; Er, Nuray; Comert, Fugen Dagli

2013-09-01

To compare the smear layer removal efficacy and erosive effects of different irrigation protocols under clinical and laboratory conditions. Mandibular third molars (n = 32) of 30-45 year-old patients were instrumented with rotary files and were randomly assigned to one of the following groups for final irrigation: (1) 5.25% NaOCl; (2) 17% EDTA; and (3) BioPure MTAD. Thereafter, the teeth were immediately extracted and processed for micromorphological investigation. In vitro specimen pairs were prepared by repeating the clinical experiments on freshly-extracted mandibular third molars. To compare open and closed systems, laboratory experiments were repeated on 32 additional teeth with enlarged apical foramen. The cleanliness of the root canals and the extent of erosion were assessed by environmental scanning electron microscopy. Specimens prepared under clinical and laboratory conditions had similar cleanliness and erosion scores (p > 0.05). Under both conditions, the tested solutions were more effective in removing the smear layer in the coronal and middle regions than in the apical one. Comparison of closed and open systems showed similar levels of cleanliness and erosion in all regions (p > 0.05), with the exception of 17% EDTA showing significantly higher levels of cleanliness and erosion in the apical third of open-end specimens. Based on clinical correlates of in vitro root canal cleanliness and erosion, laboratory testing of root canal irrigants on extracted teeth with closed apices can serve as a reliable method to simulate the clinical condition. EDTA was the most effective final irrigation solution in removing the smear layer at the expense of yielding the greatest erosive effect.

Conditioning Factors of an Organizational Learning Culture

ERIC Educational Resources Information Center

Rebelo, Teresa Manuela; Gomes, Adelino Duarte

2011-01-01

Purpose: The aim of this study is to assess the relationship between some variables (organizational structure, organizational dimension and age, human resource characteristics, the external environment, strategy and quality) and organizational learning culture and evaluate the way they interact with this kind of culture.…

Effects of culture conditions and biofilm formation on the iodine susceptibility of Legionella pneumophila

NASA Technical Reports Server (NTRS)

Cargill, K. L.; Pyle, B. H.; Sauer, R. L.; McFeters, G. A.

1992-01-01

The susceptibility of Legionella pneumophila to iodination was studied with cultures grown in well water, on rich agar media, and attached to stainless-steel surfaces. Legionella pneumophila grown in water cultures in association with other microorganisms were less sensitive to disinfection by chlorine and iodine than were agar-passaged cultures. Differences in sensitivity to disinfection between water-cultured and agar-grown legionellae were determined by comparing C x T values (concentration in milligrams per litre multiplied by time in minutes to achieve 99% decrease in viability) and CM x T values (concentration in molarity). Iodine (1500x) gave a greater difference in CM x T values than did chlorine (68x). Iodine was 50 times more effective than chlorine when used with agar-grown cultures but was only twice as effective when tested against water-grown Legionella cultures. C x T x S values (C x T multiplied by percent survivors), which take into consideration the percent surviving bacteria, were used to compare sensitivities in very resistant populations, such as those in biofilms. Water cultures of legionellae associated with stainless-steel surfaces were 135 times more resistant to iodination than were unattached legionellae, and they were 210,000 times more resistant than were agar-grown cultures. These results indicate that the conditions under which legionellae are grown can dramatically affect their susceptibility to some disinfectants and must be considered when evaluating the efficacy of a disinfecting agent.

Chromium picolinate positively influences the glucose transporter system via affecting cholesterol homeostasis in adipocytes cultured under hyperglycemic diabetic conditions

PubMed Central

Pattar, Guruprasad R.; Tackett, Lixuan; Liu, Ping; Elmendorf, Jeffrey S.

2008-01-01

Since trivalent chromium (Cr3+) enhances glucose metabolism, interest in the use of Cr3+as a therapy for type 2 diabetes has grown in the mainstream medical community. Moreover, accumulating evidence suggests that Cr3+ may also benefit cardiovascular disease (CVD) and atypical depression. We have found that cholesterol, a lipid implicated in both CVD and neurodegenerative disorders, also influences cellular glucose uptake. A recent study in our laboratory shows that exposure of 3T3-L1 adipocytes to chromium picolinate (CrPic, 10 nM) induces a loss of plasma membrane cholesterol. Concomitantly, accumulation of intracellularly sequestered glucose transporter GLUT4 at the plasma membrane was dependent on the CrPic-induced cholesterol loss. Since CrPic supplementation has the greatest benefit on glucose metabolism in hyperglycemic insulin-resistant individuals, we asked here if the CrPic effect on cells was glucose-dependent. We found that GLUT4 redistribution in cells treated with CrPic occurs only in cells cultured under high glucose (25 mM) conditions that resemble the diabetic-state, and not in cells cultured under non-diabetic (5.5 mM glucose) conditions. Examination of the effect of CrPic on proteins involved in cholesterol homeostasis revealed that the activity of sterol regulatory element-binding protein (SREBP), a membrane-bound transcription factor ultimately responsible for controlling cellular cholesterol balance, was upregulated by CrPic. In addition, ABCA1, a major player in mediating cholesterol efflux was decreased, consistent with SREBP transcriptional repression of the ABCA1 gene. Although the exact mechanism of Cr3+-induced cholesterol loss remains to be determined, these cellular responses highlight a novel and significant effect of chromium on cholesterol homeostasis. Furthermore, these findings provide an important clue to our understanding of how chromium supplementation might benefit hypercholesterolemia-associated disorders. PMID:16870493

Chromium picolinate positively influences the glucose transporter system via affecting cholesterol homeostasis in adipocytes cultured under hyperglycemic diabetic conditions.

PubMed

Pattar, Guruprasad R; Tackett, Lixuan; Liu, Ping; Elmendorf, Jeffrey S

2006-11-07

Since trivalent chromium (Cr(3+)) enhances glucose metabolism, interest in the use of Cr(3+)as a therapy for type 2 diabetes has grown in the mainstream medical community. Moreover, accumulating evidence suggests that Cr(3+) may also benefit cardiovascular disease (CVD) and atypical depression. We have found that cholesterol, a lipid implicated in both CVD and neurodegenerative disorders, also influences cellular glucose uptake. A recent study in our laboratory shows that exposure of 3T3-L1 adipocytes to chromium picolinate (CrPic, 10 nM) induces a loss of plasma membrane cholesterol. Concomitantly, accumulation of intracellularly sequestered glucose transporter GLUT4 at the plasma membrane was dependent on the CrPic-induced cholesterol loss. Since CrPic supplementation has the greatest benefit on glucose metabolism in hyperglycemic insulin-resistant individuals, we asked here if the CrPic effect on cells was glucose-dependent. We found that GLUT4 redistribution in cells treated with CrPic occurs only in cells cultured under high glucose (25 mM) conditions that resemble the diabetic-state, and not in cells cultured under non-diabetic (5.5 mM glucose) conditions. Examination of the effect of CrPic on proteins involved in cholesterol homeostasis revealed that the activity of sterol regulatory element-binding protein (SREBP), a membrane-bound transcription factor ultimately responsible for controlling cellular cholesterol balance, was upregulated by CrPic. In addition, ABCA1, a major player in mediating cholesterol efflux was decreased, consistent with SREBP transcriptional repression of the ABCA1 gene. Although the exact mechanism of Cr(3+)-induced cholesterol loss remains to be determined, these cellular responses highlight a novel and significant effect of chromium on cholesterol homeostasis. Furthermore, these findings provide an important clue to our understanding of how chromium supplementation might benefit hypercholesterolemia-associated disorders.

Expression and Secretion of Endostar Protein by Escherichia Coli: Optimization of Culture Conditions Using the Response Surface Methodology.

PubMed

Mohajeri, Abbas; Abdolalizadeh, Jalal; Pilehvar-Soltanahmadi, Younes; Kiafar, Farhad; Zarghami, Nosratollah

2016-10-01

Endostar as a specific drug in treatment of the nonsmall cell lung cancer is produced using Escherichia coli expression system. Plackett-Burman design (PBD) and response surface methodology (RSM) are statistical tools for experimental design and optimization of biotechnological processes. This investigation aimed to predict and develop the optimal culture condition and its components for expression and secretion of endostar into the culture medium of E. coli. The synthetic endostar coding sequence was fused with PhoA signal peptide. The nine factors involved in the production of recombinant protein-postinduction temperature, cell density, rotation speed, postinduction time, concentration of glycerol, IPTG, peptone, glycine, and triton X-100-were evaluated using PBD. Four significant factors were selected based on PBD results for optimizing culture condition using RSM. Endostar was purified using cation exchange chromatography and size exclusion chromatography. The maximum level of endostar was obtained under the following condition: 13.57-h postinduction time, 0.76 % glycine, 0.7 % triton X-100, and 4.87 % glycerol. The predicted levels of endostar was significantly correlated with experimental levels (R 2 = 0.982, P = 0.00). The obtained results indicated that PBD and RSM are effective tools for optimization of culture condition and its components for endostar production in E. coli. The most important factors in the enhancement of the protein production are glycerol, glycine, and postinduction time.

Idaho National Laboratory Cultural Resource Monitoring Report for FY 2008

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

Brenda R. Pace

2009-01-01

This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during fiscal year 2008 (FY 2008). Throughout the year, 45 cultural resource localities were revisited including: two locations of heightened Shoshone-Bannock tribal sensitivity, four caves, one butte, twenty-eight prehistoric archaeological sites, three historic homesteads, two historic stage stations, one historic canal construction camp, three historic trails, and Experimental Breeder Reactor-I, which is a designated National Historic Landmark. Several INL project areas were also monitored in FY 2008 to assess project compliance with cultural resource recommendations, confirm the locations of previously recordedmore » cultural resources in relation to project activities, to assess the damage caused by fire-fighting efforts, and to watch for cultural materials during ground disturbing activities. Although impacts were documented at a few locations, no significant adverse effects that would threaten the National Register eligibility of any resource were observed. Monitoring also demonstrated that INL projects generally remain in compliance with recommendations to protect cultural resources« less

Evaluation of Mycology Laboratory Proficiency Testing

PubMed Central

Reilly, Andrew A.; Salkin, Ira F.; McGinnis, Michael R.; Gromadzki, Sally; Pasarell, Lester; Kemna, Maggi; Higgins, Nancy; Salfinger, Max

1999-01-01

Changes over the last decade in overt proficiency testing (OPT) regulations have been ostensibly directed at improving laboratory performance on patient samples. However, the overt (unblinded) format of the tests and regulatory penalties associated with incorrect values allow and encourage laboratorians to take extra precautions with OPT analytes. As a result OPT may measure optimal laboratory performance instead of the intended target of typical performance attained during routine patient testing. This study addresses this issue by evaluating medical mycology OPT and comparing its fungal specimen identification error rates to those obtained in a covert (blinded) proficiency testing (CPT) program. Identifications from 188 laboratories participating in the New York State mycology OPT from 1982 to 1994 were compared with the identifications of the same fungi recovered from patient specimens in 1989 and 1994 as part of the routine procedures of 88 of these laboratories. The consistency in the identification of OPT specimens was sufficient to make accurate predictions of OPT error rates. However, while the error rates in OPT and CPT were similar for Candida albicans, significantly higher error rates were found in CPT for Candida tropicalis, Candida glabrata, and other common pathogenic fungi. These differences may, in part, be due to OPT’s use of ideal organism representatives cultured under optimum growth conditions. This difference, as well as the organism-dependent error rate differences, reflects the limitations of OPT as a means of assessing the quality of routine laboratory performance in medical mycology. PMID:10364601

Optimization of culture condition for ACEI and GABA production by lactic acid bacteria.

PubMed

Tung, Yi-Ting; Lee, Bao-Hong; Liu, Chin-Feng; Pan, Tzu-Ming

2011-01-01

Gamma-aminobutyric acid (GABA) and angiotensin-converting enzyme inhibitor (ACEI) are compounds which can influence hypertension. The goal of this study is to optimize the culture condition for GABA and ACEI production by Lactobacillus plantarum NTU 102 fermented skim milk. In this study, we used 3-factor-3-level Box-Behnken design combining with response surface methodology, where the 3 factors represent the concentration of skim milk, the concentration of monosodium glutamate, and culture temperature. Best conditions for GABA and ACEI production differed. The results indicated that L. plantarum NTU 102 produced the highest combined levels of GABA and ACEI at 37 °C, in milk having 8% to 12% nonfat solids supplemented with 0.6% to 1% MSG. Agitation of the medium during fermentation had no effect on GABA or ACEI production but extended incubation (up to 6 d) increases levels of the bioactive compounds. L. plantarum NTU 102 fermented products may be a potential functional food source for regulating hypertension. © 2011 Institute of Food Technologists®

Apoptosis as the focus of an authentic research experience in a cell physiology laboratory.

PubMed

Byrd, Shere K

2016-06-01

Curriculum-embedded independent research is a high-impact teaching practice that has been shown to increase student engagement and learning. This article describes a multiweek laboratory project for an upper-division undergraduate cell physiology laboratory using apoptosis via the mitochondrial pathway as the overarching theme. Students did literature research on apoptotic agents that acted via the mitochondrial pathway. Compounds ranged from natural products such as curcumin to synthetic compounds such as etoposide. Groups of two to three students planned a series of experiments using one of three cultured cell lines that required them to 1) learn to culture cells; 2) determine treatment conditions, including apoptotic agent solubility and concentration ranges that had been reported in the literature; 3) choose two methods to validate/quantify apoptotic capacity of the reagent; and 4) attempt to "rescue" cells from undergoing apoptosis using one of several available compounds/methods. In essence, given some reagent and equipment constraints, students designed an independent experiment to highlight the effects of different apoptotic agents on cells in culture. Students presented their experimental designs as in a laboratory group meeting and their final findings as a classroom "symposium." This exercise can be adapted to many different types of laboratories with greater or lesser equipment and instrumentation constraints, incorporates several core cell physiology methods, and encourages key experimental design and critical thinking components of independent research. Copyright © 2016 The American Physiological Society.

What are the critical steps in processing blood cultures? A prospective audit evaluating current practice of reporting blood cultures in a centralised laboratory serving secondary care hospitals.

PubMed

Meda, Manjula; Clayton, James; Varghese, Reela; Rangaiah, Jayakeerthi; Grundy, Clive; Dashti, Farnaz; Garner, David; Groves, Katherine; Fitzmaurice, Karen; Hutley, E

2017-04-01

To assess current procedures of processing positive blood cultures against national standards with an aim to evaluate its clinical impact and to determine the utility of currently available rapid identification and susceptibility tests in processing of blood cultures. Blood cultures from three secondary care hospitals, processed at a centralised laboratory, were prospectively audited. Data regarding processing times, communication with prescribers, changes to patient management and mortality within 30 days of a significant blood culture were collected in a preplanned pro forma for a 4-week period. Of 2206 blood cultures, 211 positive blood cultures flagged positive. Sixty-nine (3.1%) of all cultures were considered to be contaminated. Fifty per cent of blood cultures that flagged positive had a Gram stain reported within 2 hours. Two (0.99%) patients with a significant bacteraemia had escalation of antimicrobial treatment at the point of reporting the Gram stain that was subsequently deemed necessary once sensitivity results were known. Most common intervention was de-escalation of therapy for Gram-positive organisms at the point of availability of pathogen identification (25.6% in Gram positive vs 10% in Gram negative; p=0.012). For Gram-negative organisms, the most common intervention was de-escalation of therapy at the point of availability of sensitivity results (43% in Gram negatives vs 17.9% in Gram positive; p=0.0097). Overall mortality within 30 days of a positive blood culture was 10.9% (23/211). Antibiotic resistance may have contributed to mortality in four of these patients (three Gram negative and one Gram positive). Gram stain result had the least impact on antibiotic treatment interventions (escalation or de-escalation). Tests that improve identification time for Gram-positive pathogens and sensitivity time for Gram-negative pathogens had the greatest impact in making significant changes to antimicrobial treatment. Published by the BMJ

Condition and biochemical profile of blue mussels (Mytilus edulis L.) cultured at different depths in a cold water coastal environment

NASA Astrophysics Data System (ADS)

Gallardi, Daria; Mills, Terry; Donnet, Sebastien; Parrish, Christopher C.; Murray, Harry M.

2017-08-01

The growth and health of cultured blue mussels (Mytilus edulis) are affected by environmental conditions. Typically, culture sites are situated in sheltered areas near shore (i.e., < 1 km distance from land, < 20 m depth); however, land runoff, user conflicts and environmental impact in coastal areas are concerns and interest in developing deep water (> 20 m depth) mussel culture has been growing. This study evaluated the effect of culture depth on blue mussels in a cold water coastal environment (Newfoundland, Canada). Culture depth was examined over two years from September 2012 to September 2014; mussels from three shallow water (5 m) and three deep water (15 m) sites were compared for growth and biochemical composition; culture depths were compared for temperature and chlorophyll a. Differences between the two years examined were noted, possibly due to harsh winter conditions in the second year of the experiment. In both years shallow and deep water mussels presented similar condition; in year 2 deep water mussels had a significantly better biochemical profile. Lipid and glycogen analyses showed seasonal variations, but no significant differences between shallow and deep water were noted. Fatty acid profiles showed a significantly higher content of omega-3 s (20:5ω3; EPA) and lower content of bacterial fatty acids in deep water sites in year 2. Everything considered, deep water appeared to provide a more favorable environment for mussel growth than shallow water under harsher weather conditions.

[Culture conditions for gametes and embryos: Which culture medium? Which impact on newborn?

PubMed

Koscinski, I; Merten, M; Kazdar, N; Guéant, J-L

2018-05-01

Many studies have examined the impact of cell/embryo culture media on the development of human embryo during IVF process, but few studies have followed up and compared the effects of these culture media on the developmental outcome of children conceived by IVF. As recurrent experimental evidence from animal studies suggests potential long-term effects of embryo culture media on the health outcome of IVF-conceived children, more studies are needed to clarify the role of the culture media and mechanisms underlying such effects. In human, however, the effects of culture media are difficult to pinpoint due to complications stem from both the influence of maternal nutrition during the gestational period and the parental genetic. Based on a simple review of the literature integrating animal experimentations and human clinic studies, we suggest that the composition of culture medium should be considered beyond the character of unique or sequential medium, corresponding to "let embryo choose" or "back to nature" respectively. Instead, we suggest that the main components of embryo culture media should be considered from the point of view of metabolic consequences and potential epigenetic effects. Given that energetic metabolites can regulate epigenetic machinery, we hypothesize that metabolic abnormalities linked to morphological abnormalities could reveal epigenetic defects in embryos. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Transformation of proanthocyanidin A2 to its isomers under different physiological pH conditions and common cell culture medium.

PubMed

Lu, Wen-Chien; Huang, Wei-Ting; Kumaran, Alaganandam; Ho, Chi-Tang; Hwang, Lucy Sun

2011-06-08

Proanthocyanidins constitute an important class of polyphenols ubiquitously found in plants. They have been extensively studied for their antioxidant capacity and bioactivity in vitro and in animal models. However, their stability under different pH conditions and in cell culture medium has not been well documented. In the present study, it was observed that proanthocyanidin A2 (PA2) was relatively more stable in acidic condition than in weak alkaline condition. PA2 was also quite unstable in basal-Dulbecco's Modified Eagle medium (b-DMEM medium) at 37 °C. The addition of PA2 to the cell culture medium accelerated its epimerization with a half-life of <15 min, and ethylenediaminetetraacetic acid (EDTA) could not stop the reaction. The results also demonstrated that the major isomers transformed in the weak alkaline condition or cell culture medium at 37 °C were identified as epicatechin-(4β→8; 2β→O→7)-ent-catechin (proanthocyanidin A4) and epicatechin-(4β→6; 2β→O→7)-ent-catechin. The rates of transformation were dependent on the pH or the components of the medium. Therefore, the results obtained for PA2 in the cell culture bioassays, which were usually carried out for 24 h, might not represent the true activity of the original PA2. The stability and transformation of PA2 should be considered when the bioactivity of PA2 is evaluated in a given cell culture system.

Low-Oxygen Culture Conditions Extend the Multipotent Properties of Human Retinal Progenitor Cells

PubMed Central

Tucker, Budd A.; Young, Michael J.

2014-01-01

Purpose: Development of an effective cell-based therapy is highly dependent upon having a reproducible cell source suitable for transplantation. One potential source, isolated from the developing fetal neural retina, is the human retinal progenitor cell (hRPC). One limiting factor for the use of hRPCs is their in vitro expansion limit. As such, the aim of this study was to determine whether culturing hRPCs under 3% O2 would support their proliferative capacity while maintaining multipotency. Methods: To determine the effect of low oxygen on the ability of hRPCs to self-renew, rates of proliferation and apoptosis, telomerase activity, and expression of proliferative, stemness, and differentiation markers were assessed for hRPCs cultured in 3% and 20% oxygen conditions. Results: Culture under 3% oxygen increases the proliferation rate and shifts the proliferation limit of hRPCs to greater 40 divisions. This increased capacity for proliferation is correlated with an upregulation of Ki67, CyclinD1, and telomerase activity and a decrease in p53 expression and apoptosis. Increased expression of cMyc, Klf4, Oct4, and Sox2 in 3% O2 is correlated with stabilization of both HIF1α and HIF2α. The eye field development markers Pax6, Sox2, and Otx2 are present in hRPCs up to passage 16 in 3% O2. Following in vitro differentiation hRPCs expanded in the 3% O2 were able to generate specialized retinal cells, including rods and cones. Conclusions: Low-oxygen culture conditions act to maintain both multipotency and self-renewal properties of hRPCs in vitro. The extended expansion limits permit the development of a clinical-grade reagent for transplantation. PMID:24320879

Performance of Gram staining on blood cultures flagged negative by an automated blood culture system.

PubMed

Peretz, A; Isakovich, N; Pastukh, N; Koifman, A; Glyatman, T; Brodsky, D

2015-08-01

Blood is one of the most important specimens sent to a microbiology laboratory for culture. Most blood cultures are incubated for 5-7 days, except in cases where there is a suspicion of infection caused by microorganisms that proliferate slowly, or infections expressed by a small number of bacteria in the bloodstream. Therefore, at the end of incubation, misidentification of positive cultures and false-negative results are a real possibility. The aim of this work was to perform a confirmation by Gram staining of the lack of any microorganisms in blood cultures that were identified as negative by the BACTEC™ FX system at the end of incubation. All bottles defined as negative by the BACTEC FX system were Gram-stained using an automatic device and inoculated on solid growth media. In our work, 15 cultures that were defined as negative by the BACTEC FX system at the end of the incubation were found to contain microorganisms when Gram-stained. The main characteristic of most bacteria and fungi growing in the culture bottles that were defined as negative was slow growth. This finding raises a problematic issue concerning the need to perform Gram staining of all blood cultures, which could overload the routine laboratory work, especially laboratories serving large medical centers and receiving a large number of blood cultures.

Medium components and culture conditions affect the thermotolerance of aerial conidia of fungal biocontrol agent Beauveria bassiana.

PubMed

Ying, S-H; Feng, M-G

2006-09-01

To produce more thermotolerable conidia of Beauveria bassiana, a well-known fungal biocontrol agent, by optimizing the medium components and culture conditions. The conidia produced on media including 0.5-6% glucose, sucrose or starch as carbon source and 50-300-microg ml(-1) Cu2+, Zn2+, Mn2+ or Fe3+ as additive to Sabouraud dextrose medium at 15-30 degrees C, pH 4-8 or KCl-adjusted water availabilities were exposed to 30-min wet heat stress at 48 degrees C. The medium components for conidial production with greatly enhanced thermotolerance included 4% glucose as optimum or 1% starch as alternative for the carbon source and < or =50-microg ml(-1) Mn2+ for the metal additive. The culture conditions were optimized as 25 degrees C and pH 5-6. Conidial thermotolerance decreased remarkably when sucrose and Fe3+ or Cu2+ were used in the cultures, but altered slightly when 50-200-microg ml(-1) Zn2+ were included. The tolerance of B. bassiana conidia to the thermal stress was significantly affected by the medium composition and culture conditions under which the conidia were produced. Proper treatment of small grains as mass production substrates for more glucose release and supplement of glucose or 50-microg ml(-1) Mn2+ are possible means to enhancing conidial thermotolerance and field persistence for improved insect control.

The Naval Health Research Center Respiratory Disease Laboratory.

PubMed

Ryan, M; Gray, G; Hawksworth, A; Malasig, M; Hudspeth, M; Poddar, S

2000-07-01

Concern about emerging and reemerging respiratory pathogens prompted the development of a respiratory disease reference laboratory at the Naval Health Research Center. Professionals working in this laboratory have instituted population-based surveillance for pathogens that affect military trainees and responded to threats of increased respiratory disease among high-risk military groups. Capabilities of this laboratory that are unique within the Department of Defense include adenovirus testing by viral shell culture and microneutralization serotyping, influenza culture and hemagglutination inhibition serotyping, and other special testing for Streptococcus pneumoniae, Streptococcus pyogenes, Mycoplasma pneumonia, and Chlamydia pneumoniae. Projected capabilities of this laboratory include more advanced testing for these pathogens and testing for other emerging pathogens, including Bordetella pertussis, Legionella pneumoniae, and Haemophilus influenzae type B. Such capabilities make the laboratory a valuable resource for military public health.

Individuality, phenotypic differentiation, dormancy and ‘persistence’ in culturable bacterial systems: commonalities shared by environmental, laboratory, and clinical microbiology

PubMed Central

Kell, Douglas; Potgieter, Marnie; Pretorius, Etheresia

2015-01-01

For bacteria, replication mainly involves growth by binary fission. However, in a very great many natural environments there are examples of phenotypically dormant, non-growing cells that do not replicate immediately and that are phenotypically ‘nonculturable’ on media that normally admit their growth. They thereby evade detection by conventional culture-based methods. Such dormant cells may also be observed in laboratory cultures and in clinical microbiology. They are usually more tolerant to stresses such as antibiotics, and in clinical microbiology they are typically referred to as ‘persisters’. Bacterial cultures necessarily share a great deal of relatedness, and inclusive fitness theory implies that there are conceptual evolutionary advantages in trading a variation in growth rate against its mean, equivalent to hedging one’s bets. There is much evidence that bacteria exploit this strategy widely. We here bring together data that show the commonality of these phenomena across environmental, laboratory and clinical microbiology. Considerable evidence, using methods similar to those common in environmental microbiology, now suggests that many supposedly non-communicable, chronic and inflammatory diseases are exacerbated (if not indeed largely caused) by the presence of dormant or persistent bacteria (the ability of whose components to cause inflammation is well known). This dormancy (and resuscitation therefrom) often reflects the extent of the availability of free iron. Together, these phenomena can provide a ready explanation for the continuing inflammation common to such chronic diseases and its correlation with iron dysregulation. This implies that measures designed to assess and to inhibit or remove such organisms (or their access to iron) might be of much therapeutic benefit. PMID:26629334

Laboratory Evaluation and Application of Microwave Absorption Properties under Simulated Conditions for Planetary Atmospheres

NASA Technical Reports Server (NTRS)

Steffes, Paul G.

2002-01-01

Radio absorptivity data for planetary atmospheres obtained from spacecraft radio occultation experiments, entry probe radio signal absorption measurements, and earth-based or spacecraft-based radio astronomical (emission) observations can be used to infer abundances of microwave absorbing constituents in those atmospheres, as long as reliable information regarding the microwave absorbing properties of potential constituents is available. The use of theoretically-derived microwave absorption properties for such atmospheric constituents, or the use of laboratory measurements of such properties taken under environmental conditions that are significantly different than those of the planetary atmosphere being studied, often leads to significant misinterpretation of available opacity data. Laboratory measurements have shown that the centimeter-wavelength opacity from gaseous phosphine (PH3) under simulated conditions for the outer planets far exceeds that predicted from theory over a wide range of temperatures and pressures. This fundamentally changed the resulting interpretation of Voyager radio occultation data at Saturn and Neptune. It also directly impacts planning and scientific goals for study of Saturn's atmosphere with the Cassini Radio Science Experiment and the Rossini RADAR instrument. The recognition of the need to make such laboratory measurements of simulated planetary atmospheres over a range of temperatures and pressures which correspond to the altitudes probed by both radio occultation experiments and radio astronomical observations, and over a range of frequencies which correspond to those used in both spacecraft entry probe and orbiter (or flyby) radio occultation experiments and radio astronomical observations, has led to the development of a facility at Georgia Tech which is capable of making such measurements. It has been the goal of this investigation to conduct such measurements and to apply the results to a wide range of planetary observations

Algal recycling enhances algal productivity and settleability in Pediastrum boryanum pure cultures.

PubMed

Park, Jason B K; Craggs, Rupert J; Shilton, Andy N

2015-12-15

Recycling a portion of gravity harvested algae (i.e. algae and associated bacteria biomass) has been shown to improve both algal biomass productivity and harvest efficiency by maintaining the dominance of a rapidly-settleable colonial alga, Pediastrum boryanum in both pilot-scale wastewater treatment High Rate Algal Ponds (HRAP) and outdoor mesocosms. While algal recycling did not change the relative proportions of algae and bacteria in the HRAP culture, the contribution of the wastewater bacteria to the improved algal biomass productivity and settleability with the recycling was not certain and still required investigation. P. boryanum was therefore isolated from the HRAP and grown in pure culture on synthetic wastewater growth media under laboratory conditions. The influence of recycling on the productivity and settleability of the pure P. boryanum culture was then determined without wastewater bacteria present. Six 1 L P. boryanum cultures were grown over 30 days in a laboratory growth chamber simulating New Zealand summer conditions either with (Pr) or without (Pc) recycling of 10% of gravity harvested algae. The cultures with recycling (Pr) had higher algal productivity than the controls (Pc) when the cultures were operated at both 4 and 3 d hydraulic retention times by 11% and 38% respectively. Furthermore, algal recycling also improved 1 h settleability from ∼60% to ∼85% by increasing the average P. boryanum colony size due to the extended mean cell residence time and promoted formation of large algal bio-flocs (>500 μm diameter). These results demonstrate that the presence of wastewater bacteria was not necessary to improve algal productivity and settleability with algal recycling. Copyright © 2015 Elsevier Ltd. All rights reserved.

Importance of supply integrity for in vitro fertilization and embryo culture.

PubMed

Morbeck, Dean E

2012-06-01

The quality of in vitro culture conditions is a key component of a successful clinical embryology laboratory. Many, but not all, supplies used in the embryology laboratory are screened by the supplier with a bioassay. Embryology laboratories use a variety of approaches to verify the quality of mineral oil, protein, and disposables before clinical use; however, a best practice has not been determined. Some laboratories test every supply, even those already screened by the supplier, whereas other laboratories perform as little testing as possible. Despite screening by the supplier, recent reports of embryo toxicity, specifically with mineral oil, highlight that the integrity of the supply system has gaps. This review describes current bioassay quality control testing and discusses how it applies to screening of products with documented lot-to-lot variation. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Virtual Cultural Landscape Laboratory Based on Internet GIS Technology

NASA Astrophysics Data System (ADS)

Bill, R.

2012-07-01

In recent years the transfer of old documents (books, paintings, maps etc.) from analogue to digital form has gained enormous importance. Numerous interventions are concentrated in the digitalisation of library collections, but also commercial companies like Microsoft or Google try to convert large analogue stocks such as books, paintings, etc. in digital form. Data in digital form can be much easier made accessible to a large user community, especially to the interested scientific community. The aim of the described research project is to set up a virtual research environment for interdisciplinary research focusing on the landscape of the historical Mecklenburg in the north-east of Germany. Georeferenced old maps from 1786 and 1890 covering complete Mecklenburg should be combined with current geo-information, satellite and aerial imagery to support spatio-temporal research aspects in different scales in space (regional 1:200,000 to local 1:25.000) and time (nearly 250 years in three time steps, the last 30 years also in three time slices). The Virtual Laboratory for Cultural Landscape Research (VKLandLab) is designed and developed by the Chair of Geodesy and Geoinformatics, hosted at the Computing Centre (ITMZ) and linked to the Digital Library (UB) at Rostock University. VKLandLab includes new developments such as wikis, blogs, data tagging, etc. and proven components already integrated in various data-related infrastructures such as InternetGIS, data repositories and authentication structures. The focus is to build a data-related infrastructure and a work platform that supports students as well as researchers from different disciplines in their research in space and time.

Stability of spinosad resistance in Frankliniella occidentalis (Pergande) under laboratory conditions.

PubMed

Bielza, P; Quinto, V; Grávalos, C; Fernández, E; Abellán, J; Contreras, J

2008-08-01

The stability of spinosad resistance in western flower thrips (WFT), Frankliniella occidentalis (Pergande), populations with differing initial frequencies of resistance was studied in laboratory conditions. The stability of resistance was assessed in bimonthly residual bioassays in five populations with initial frequencies of 100, 75, 50, 25 and 0% of resistant individuals. There were no consistent changes in susceptibility of the susceptible strain after eight months without insecticide pressure. In the resistant strain, very highly resistant to spinosad (RF50>23,000-fold), resistance was maintained up to eight months without further exposure to spinosad. In the absence of any immigration of susceptible genes into the population, resistance was stable. In the case of the population with different initial frequency of resistant thrips, spinosad resistance declined significantly two months later in the absence of selection pressure. With successive generations, these strains did not change significantly in sensitivity. Spinosad resistance in F. occidentalis declined significantly in the absence of selection pressure and the presence of susceptible WFT. These results suggest that spinosad resistance probably is unstable under field conditions, primarily due to the immigration of susceptible WFT. Factors influencing stability or reversion of spinosad resistance are discussed.

Elimination of remaining undifferentiated induced pluripotent stem cells in the process of human cardiac cell sheet fabrication using a methionine-free culture condition.

PubMed

Matsuura, Katsuhisa; Kodama, Fumiko; Sugiyama, Kasumi; Shimizu, Tatsuya; Hagiwara, Nobuhisa; Okano, Teruo

2015-03-01

Cardiac tissue engineering is a promising method for regenerative medicine. Although we have developed human cardiac cell sheets by integration of cell sheet-based tissue engineering and scalable bioreactor culture, the risk of contamination by induced pluripotent stem (iPS) cells in cardiac cell sheets remains unresolved. In the present study, we established a novel culture method to fabricate human cardiac cell sheets with a decreased risk of iPS cell contamination while maintaining viabilities of iPS cell-derived cells, including cardiomyocytes and fibroblasts, using a methionine-free culture condition. When cultured in the methionine-free condition, human iPS cells did not survive without feeder cells and could not proliferate or form colonies on feeder cells or in coculture with cells for cardiac cell sheet fabrication. When iPS cell-derived cells after the cardiac differentiation were transiently cultured in the methionine-free condition, gene expression of OCT3/4 and NANOG was downregulated significantly compared with that in the standard culture condition. Furthermore, in fabricated cardiac cell sheets, spontaneous and synchronous beating was observed in the whole area while maintaining or upregulating the expression of various cardiac and extracellular matrix genes. These findings suggest that human iPS cells are methionine dependent and a methionine-free culture condition for cardiac cell sheet fabrication might reduce the risk of iPS cell contamination.

Optimization of the conditions of isolation and culture of dairy goat male germline stem cells (mGSC).

PubMed

Zhu, Haijing; Liu, Chao; Li, Mingzhao; Sun, Junwei; Song, Wencong; Hua, Jinlian

2013-02-01

Male germline stem cells (mGSC) reside in the basement of seminiferous tubules of the testis and have the capacity of self-renewal and differentiation into sperm throughout the life of animals. Reports on mice and human mGSC have demonstrated that mGSC are an unlimited resource of pluripotent stem cells for sperm production. The conditions of isolation and culture of mouse and human mGSC are well developed; however, the systematic culture conditions of dairy goat mGSC are still deficient although there have been several reports of successful cultures. With the present research, several key elements of isolation and culture of dairy goat mGSC have been determined. Details for the conditions of isolation of dairy testicular spermatogonium cells were optimized, and effects of several extracellular matrix types, ages of dairy goat, and cytokines on enrichment and culture of mGSC were compared. Biological characteristics of the cells were also evaluated by RT-PCR and immunofluorescent staining. The results indicated there is one kind of enzyme cocktail (CTHD (1mg/ml collagenase, 10μg/ml DNase, 1mg/ml hyaluronidase and 1mg/ml trypsin) combined TD (0.25% trypsin and 10mg/ml DNaseI)) that can be used to successfully isolate dairy goat testicular spermatogonium cells efficiently; and fibronectin as well as laminin were efficient extracellular matrix to enrich mGSC among the extracellular matrix types evaluated. Age of dairy goat clearly influenced the cultures of dairy goat mGSC with the efficiency of establishment of an mGSC line being greater if the age of the dairy goat is younger. Some cytokines e.g. BIO (A GSK3 inhibitor, 6-bromoindirubin-3'-oxime) and basic fibroblast growth factor (bFGF) acted positively on the maintenance of proliferation and pluripotency of mGSC. Leukemia inhibitory factor (LIF) might, however, inhibit the proliferation of dairy goat mGSC. These cultured mGSC maintained similar characteristics as mouse and human mGSC. These results provide an

Medical Service Clinical Laboratory Procedures--Bacteriology.

ERIC Educational Resources Information Center

Department of the Army, Washington, DC.

This manual presents laboratory procedures for the differentiation and identification of disease agents from clinical materials. Included are procedures for the collection of specimens, preparation of culture media, pure culture methods, cultivation of the microorganisms in natural and simulated natural environments, and procedures in…

Identification of a New Marine Bacterial Strain SD8 and Optimization of Its Culture Conditions for Producing Alkaline Protease

PubMed Central

Cui, Hongxia; Yang, Muyang; Wang, Liping; Xian, Cory J.

2015-01-01

While much attention has been given to marine microorganisms for production of enzymes, which in general are relatively more stable and active compared to those from plants and animals, studies on alkaline protease production from marine microorganisms have been very limited. In the present study, the alkaline protease producing marine bacterial strain SD8 isolated from sea muds in the Geziwo Qinhuangdao sea area of China was characterized and its optimal culture conditions were investigated. Strain SD8 was initially classified to belong to genus Pseudomonas by morphological, physiological and biochemical characterizations, and then through 16S rDNA sequence it was identified to be likely Pseudomonas hibiscicola. In addition, the culture mediums, carbon sources and culture conditions of strain SD8 were optimized for maximum production of alkaline protease. Optimum enzyme production (236U/mL when cultured bacteria being at 0.75 mg dry weight/mL fermentation broth) was obtained when the isolate at a 3% inoculum size was grown in LB medium at 20 mL medium/100mL Erlenmeyer flask for 48h culture at 30°C with an initial of pH 7.5. This was the first report of strain Pseudomonas hibiscicola secreting alkaline protease, and the data for its optimal cultural conditions for alkaline protease production has laid a foundation for future exploration for the potential use of SD8 strain for alkaline protease production. PMID:26716833

IMAGE Project: Results of Laboratory Tests on Tracers for Supercritical Conditions.

NASA Astrophysics Data System (ADS)

Brandvoll, Øyvind; Opsahl Viig, Sissel; Nardini, Isabella; Muller, Jiri

2016-04-01

The use of tracers is a well-established technique for monitoring dynamic behaviour of water and gas through a reservoir. In geothermal reservoirs special challenges are encountered due to high temperatures and pressures. In this work, tracer candidates for monitoring water at supercritical conditions (temperature > 374°C, pressure ca 218 bar), are tested in laboratory experiments. Testing of tracers at supercritical water conditions requires experimental set-ups which tolerate harsh conditions with respect to high temperature and pressure. In addition stringent HES (health, environment and safety) factors have to be taken into consideration when designing and performing the experiments. The setup constructed in this project consists of a pressure vessel, high pressure pump, instrumentation for pressure and temperature control and instrumentation required for accurate sampling of tracers. In order to achieve accurate results, a special focus has been paid to the development of the tracer sampling technique. Perfluorinated cyclic hydrocarbons (PFCs) have been selected as tracer candidates. This group of compounds is today commonly used as gas tracers in oil reservoirs. According to the literature they are stable at temperatures up to 400°C. To start with, five PFCs have been tested for thermal stability in static experiments at 375°C and 108 bar in the experimental setup described above. The tracer candidates will be further tested for several months at the relevant conditions. Preliminary results indicate that some of the PFC compounds show stability after three months. However, in order to arrive at conclusive results, the experiments have to be repeated over a longer period and paying special attention to more accurate sampling procedures.

Brain stem slice conditioned medium contains endogenous BDNF and GDNF that affect neural crest boundary cap cells in co-culture.

PubMed

Kaiser, Andreas; Kale, Ajay; Novozhilova, Ekaterina; Siratirakun, Piyaporn; Aquino, Jorge B; Thonabulsombat, Charoensri; Ernfors, Patrik; Olivius, Petri

2014-05-30

Conditioned medium (CM), made by collecting medium after a few days in cell culture and then re-using it to further stimulate other cells, is a known experimental concept since the 1950s. Our group has explored this technique to stimulate the performance of cells in culture in general, and to evaluate stem- and progenitor cell aptitude for auditory nerve repair enhancement in particular. As compared to other mediums, all primary endpoints in our published experimental settings have weighed in favor of conditioned culture medium, where we have shown that conditioned culture medium has a stimulatory effect on cell survival. In order to explore the reasons for this improved survival we set out to analyze the conditioned culture medium. We utilized ELISA kits to investigate whether brain stem (BS) slice CM contains any significant amounts of brain-derived neurotrophic factor (BDNF) and glial cell derived neurotrophic factor (GDNF). We further looked for a donor cell with progenitor characteristics that would be receptive to BDNF and GDNF. We chose the well-documented boundary cap (BC) progenitor cells to be tested in our in vitro co-culture setting together with cochlear nucleus (CN) of the BS. The results show that BS CM contains BDNF and GDNF and that survival of BC cells, as well as BC cell differentiation into neurons, were enhanced when BS CM were used. Altogether, we conclude that BC cells transplanted into a BDNF and GDNF rich environment could be suitable for treatment of a traumatized or degenerated auditory nerve. Copyright © 2014 Elsevier B.V. All rights reserved.

Cultured Human Retinal Pigment Epithelial (hRPE) Sheets: A Search for Suitable Storage Conditions.

PubMed

Khan, Ayyad Z; Utheim, Tor P; Reppe, Sjur; Sandvik, Leiv; Lyberg, Torstein; Roald, Borghild B-H; Ibrahim, Ibrahim B; Eidet, Jon R

2018-04-01

The advancement of human retinal pigment epithelial cell (hRPE) replacement therapy is partly dependent on optimization of cell culture, cell preservation, and storage medium. This study was undertaken to search for a suitable storage temperature and storage medium for hRPE. hRPE monolayer sheets were cultured under standard conditions at 37°C and then randomized for storage at six temperatures (4, 16, 20, 24, 28, and 37°C) for 7 days. After revealing a suitable storage temperature, hRPE sheets were subsequently stored with and without the silk protein sericin added to the storage medium. Live/dead assay, light microscopy, pH, and phenotypic expression of various proteins were used to assess cell cultures stored at different temperatures. After 7 days of storage, hRPE morphology was best preserved at 4°C. Addition of sericin to the storage medium maintained the characteristic morphology of the preserved cells, and improved pigmentation and levels of pigmentation-related proteins in the cultured hRPE sheets following a 7-day storage period at 4°C.

An Introductory Undergraduate Course Covering Animal Cell Culture Techniques

ERIC Educational Resources Information Center

Mozdziak, Paul E.; Petitte, James N.; Carson, Susan D.

2004-01-01

Animal cell culture is a core laboratory technique in many molecular biology, developmental biology, and biotechnology laboratories. Cell culture is a relatively old technique that has been sparingly taught at the undergraduate level. The traditional methodology for acquiring cell culture training has been through trial and error, instruction when…

Culture Conditions for Production of Biomass, Adenosine, and Cordycepin from Cordyceps sinensis CS1197: Optimization by Desirability Function Method

PubMed Central

Ghatnur, Shashidhar M.; Parvatam, Giridhar; Balaraman, Manohar

2015-01-01

Background: Cordyceps sinensis (CS) is a traditional Chinese medicine contains potent active metabolites such as nucleosides and polysaccharides. The submerged cultivation technique is studied for the large scale production of CS for biomass and metabolites production. Objective: To optimize culture conditions for large-scale production of CS1197 biomass and metabolites production. Materials and Methods: The CS1197 strain of CS was isolated from dead larvae of natural CS and the authenticity was assured by the presence of two major markers adenosine and cordycepin by high performance liquid chromatography and mass spectrometry. A three-level Box-Behnken design was employed to optimize process parameters culturing temperature, pH, and inoculum volume for the biomass yield, adenosine and cordycepin. The experimental results were regressed to a second-order polynomial equation by a multiple regression analysis for the prediction of biomass yield, adenosine and cordycepin production. Multiple responses were optimized based on desirability function method. Results: The desirability function suggested the process conditions temperature 28°C, pH 7 and inoculum volume 10% for optimal production of nutraceuticals in the biomass. The water extracts from dried CS1197 mycelia showed good inhibition for 2 diphenyl-1-picrylhydrazyl and 2,2-azinobis-(3-ethyl-benzo-thiazoline-6-sulfonic acid-free radicals. Conclusion: The result suggests that response surface methodology-desirability function coupled approach can successfully optimize the culture conditions for CS1197. SUMMARY Authentication of CS1197 strain by the presence of adenosine and cordycepin and culturing period was determined to be for 14 daysContent of nucleosides in natural CS was found higher than in cultured CS1197 myceliumBox-Behnken design to optimize critical cultural conditions: temperature, pH and inoculum volumeWater extract showed better antioxidant activity proving credible source of natural antioxidants

Three-dimensional culture conditions differentially affect astrocyte modulation of brain endothelial barrier function in response to transforming growth factor β1.

PubMed

Hawkins, Brian T; Grego, Sonia; Sellgren, Katelyn L

2015-05-22

Blood-brain barrier (BBB) function is regulated by dynamic interactions among cell types within the neurovascular unit, including astrocytes and endothelial cells. Co-culture models of the BBB typically involve astrocytes seeded on two-dimensional (2D) surfaces, which recent studies indicate cause astrocytes to express a phenotype similar to that of reactive astrocytes in situ. We hypothesized that the culture conditions of astrocytes would differentially affect their ability to modulate BBB function in vitro. Brain endothelial cells were grown alone or in co-culture with astrocytes. Astrocytes were grown either as conventional (2D) monolayers, or in a collagen-based gel which allows them to grow in a three-dimensional (3D) construct. Astrocytes were viable in 3D conditions, and displayed a marked reduction in their expression of glial fibrillary acidic protein (GFAP), suggesting reduced activation. Stimulation of astrocytes with transforming growth factor (TGF)β1 decreased transendothelial electrical resistance (TEER) and reduced expression of claudin-5 in co-cultures, whereas treatment of endothelial cells in the absence of astrocytes was without effect. The effect of TGFβ1 on TEER was significantly more pronounced in endothelial cells cultured with 3D astrocytes compared to 2D astrocytes. These results demonstrate that astrocyte culture conditions differentially affect their ability to modulate brain endothelial barrier function, and suggest a direct relationship between reactive gliosis and BBB permeability. Moreover, these studies demonstrate the potential importance of physiologically relevant culture conditions to in vitro modeling of disease processes that affect the neurovascular unit. Copyright © 2015 Elsevier B.V. All rights reserved.

Development and comparison of a real-time PCR assay for detection of Dichelobacter nodosus with culturing and conventional PCR: harmonisation between three laboratories

PubMed Central

2012-01-01

Background Ovine footrot is a contagious disease with worldwide occurrence in sheep. The main causative agent is the fastidious bacterium Dichelobacter nodosus. In Scandinavia, footrot was first diagnosed in Sweden in 2004 and later also in Norway and Denmark. Clinical examination of sheep feet is fundamental to diagnosis of footrot, but D. nodosus should also be detected to confirm the diagnosis. PCR-based detection using conventional PCR has been used at our institutes, but the method was laborious and there was a need for a faster, easier-to-interpret method. The aim of this study was to develop a TaqMan-based real-time PCR assay for detection of D. nodosus and to compare its performance with culturing and conventional PCR. Methods A D. nodosus-specific TaqMan based real-time PCR assay targeting the 16S rRNA gene was designed. The inclusivity and exclusivity (specificity) of the assay was tested using 55 bacterial and two fungal strains. To evaluate the sensitivity and harmonisation of results between different laboratories, aliquots of a single DNA preparation were analysed at three Scandinavian laboratories. The developed real-time PCR assay was compared to culturing by analysing 126 samples, and to a conventional PCR method by analysing 224 samples. A selection of PCR-products was cloned and sequenced in order to verify that they had been identified correctly. Results The developed assay had a detection limit of 3.9 fg of D. nodosus genomic DNA. This result was obtained at all three laboratories and corresponds to approximately three copies of the D. nodosus genome per reaction. The assay showed 100% inclusivity and 100% exclusivity for the strains tested. The real-time PCR assay found 54.8% more positive samples than by culturing and 8% more than conventional PCR. Conclusions The developed real-time PCR assay has good specificity and sensitivity for detection of D. nodosus, and the results are easy to interpret. The method is less time-consuming than either

Clinical and Laboratory Potential Predictors of Blood Culture Positivity in Under Five Children with Clinically Severe Pneumonia - Khartoum -Sudan.

PubMed

Salih, Karimeldin Mohamed Ali; El-Samani, El-Fatih; Bilal, Jalal Ali; Eldouch, Widad; Ibrahim, Salah Ahmed

2015-08-01

Blood culture is necessary for appropriate management of clinically severe pneumonia in children under five years of age. However, in limited resource countries it might be unduly costly and waste of valuable time because of the high negative culture rate. This study aims to identify clinical and laboratory parameters that potentially predict a positive blood culture in cases of severe pneumonia. A hospital based study, enrolled 189 cases satisfying the WHO definition of severe pneumonia. Age, gender, clinical history, physical examination, temperature, complete blood count, C-reactive protein, blood culture and Chest X Ray for all the patients were recorded. Forty one patients had positive blood culture giving a prevalence of 21.7%. All variables were used in a dichotomous manner. White Blood Count (WBC) more than 20 000, very high C-reactive protein (C-RP ≥8mg/L) and Temperature more than 40(o)C, had a positive predictive value of 46.1%, 44.3% and 40.0% respectively for a positive culture as well as a Negative Predictive Value of 91.1%, 91.6% and 91.7% respectively. The WBC more than 20 000 and temperature above 40(o)C had a significant association with a positive blood culture. Their adjusted Odds Ratios were 3.9 (95% CI: 1.4-10.90) and 3.1 (95% CI: 1.2-8.4) respectively. This was not the case for C-RP (Odds Ratio=2.2, 95% CI: 0.7-2.2) or positive Chest X Ray (Odds Ratio=1.5, 95% CI: 0.6-3.6). Temperature of more than 40(o)C, Very high C-RP and WBC of more than 20 000 are good indicators of a potential positive blood culture. It is therefore recommended that further research be undertaken to refine these predictors as screening tools before resorting to blood culture. It is also recommended that antibiotic treatment may be initiated on the basis of the high temperature and WBC, while waiting for the culture results.

The Role of Culture Theory in Cross-Cultural Training: A Multimethod Study of Culture-Specific, Culture-General, and Culture Theory-Based Assimilators.

ERIC Educational Resources Information Center

Bhawuk, Dharm P. S.

1998-01-01

In a multimethod evaluation of cross-cultural training tools involving 102 exchange students at a midwestern university, a theory-based individualism and collectivism assimilator tool had significant advantages over culture-specific and culture-general assimilators and a control condition. Results support theory-based culture assimilators. (SLD)

Surge capacity for response to bioterrorism in hospital clinical microbiology laboratories.

PubMed

Shapiro, Daniel S

2003-12-01

Surge capacity is the ability to rapidly mobilize to meet an increased demand. While large amounts of federal funding have been allocated to public health laboratories, little federal funding has been allocated to hospital microbiology laboratories. There are concerns that hospital laboratories may have inadequate surge capacities to deal with a significant bioterrorism incident. A workflow analysis of a clinical microbiology laboratory that serves an urban medical center was performed to identify barriers to surge capacity in the setting of a bioterrorism event and to identify solutions to these problems. Barriers include a national shortage of trained medical technologists, the inability of clinical laboratories to deal with a dramatic increase in the number of blood cultures, a delay while manufacturers increase production of critical products and then transport and deliver these products to clinical laboratories, and a shortage of class II biological safety cabinets. Federal funding could remedy staffing shortages by making the salaries of medical technologists comparable to those of similarly educated health care professionals and by providing financial incentives for students to enroll in clinical laboratory science programs. Blood culture bottles, and possibly continuous-monitoring blood culture instruments, should be added to the national antibiotic stockpile. Federal support must ensure that companies that manufacture essential laboratory supplies are capable of rapidly scaling up production. Hospitals must provide increased numbers of biological safety cabinets and amounts of space dedicated to clinical microbiology laboratories. Laboratories should undertake limited cross-training of technologists, ensure that adequate packaging supplies are available, and be able to move to a 4-day blood culture protocol.

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.

Finite element study of scaffold architecture design and culture conditions for tissue engineering.

PubMed

Olivares, Andy L; Marsal, Elia; Planell, Josep A; Lacroix, Damien

2009-10-01

Tissue engineering scaffolds provide temporary mechanical support for tissue regeneration and transfer global mechanical load to mechanical stimuli to cells through its architecture. In this study the interactions between scaffold pore morphology, mechanical stimuli developed at the cell microscopic level, and culture conditions applied at the macroscopic scale are studied on two regular scaffold structures. Gyroid and hexagonal scaffolds of 55% and 70% porosity were modeled in a finite element analysis and were submitted to an inlet fluid flow or compressive strain. A mechanoregulation theory based on scaffold shear strain and fluid shear stress was applied for determining the influence of each structures on the mechanical stimuli on initial conditions. Results indicate that the distribution of shear stress induced by fluid perfusion is very dependent on pore distribution within the scaffold. Gyroid architectures provide a better accessibility of the fluid than hexagonal structures. Based on the mechanoregulation theory, the differentiation process in these structures was more sensitive to inlet fluid flow than axial strain of the scaffold. This study provides a computational approach to determine the mechanical stimuli at the cellular level when cells are cultured in a bioreactor and to relate mechanical stimuli with cell differentiation.

Practical Application of Electrochemical Nitrate Sensor under Laboratory and Forest Nursery Conditions.

PubMed

Caron, William-Olivier; Lamhamedi, Mohammed S; Viens, Jeff; Messaddeq, Younès

2016-07-28

The reduction of nitrate leaching to ensure greater protection of groundwater quality has become a global issue. The development of new technologies for more accurate dosing of nitrates helps optimize fertilization programs. This paper presents the practical application of a newly developed electrochemical sensor designed for in situ quantification of nitrate. To our knowledge, this paper is the first to report the use of electrochemical impedance to determine nitrate concentrations in growing media under forest nursery conditions. Using impedance measurements, the sensor has been tested in laboratory and compared to colorimetric measurements of the nitrate. The developed sensor has been used in water-saturated growing medium and showed good correlation to certified methods, even in samples obtained over a multi-ion fertilisation season. A linear and significant relationship was observed between the resistance and the concentration of nitrates (R² = 0.972), for a range of concentrations of nitrates. We also observed stability of the sensor after exposure of one month to the real environmental conditions of the forest nursery.

Practical Application of Electrochemical Nitrate Sensor under Laboratory and Forest Nursery Conditions

PubMed Central

Caron, William-Olivier; Lamhamedi, Mohammed S.; Viens, Jeff; Messaddeq, Younès

2016-01-01

The reduction of nitrate leaching to ensure greater protection of groundwater quality has become a global issue. The development of new technologies for more accurate dosing of nitrates helps optimize fertilization programs. This paper presents the practical application of a newly developed electrochemical sensor designed for in situ quantification of nitrate. To our knowledge, this paper is the first to report the use of electrochemical impedance to determine nitrate concentrations in growing media under forest nursery conditions. Using impedance measurements, the sensor has been tested in laboratory and compared to colorimetric measurements of the nitrate. The developed sensor has been used in water-saturated growing medium and showed good correlation to certified methods, even in samples obtained over a multi-ion fertilisation season. A linear and significant relationship was observed between the resistance and the concentration of nitrates (R2 = 0.972), for a range of concentrations of nitrates. We also observed stability of the sensor after exposure of one month to the real environmental conditions of the forest nursery. PMID:27483266

Cell Culture as an Alternative in Education.

ERIC Educational Resources Information Center

Nardone, Roland M.

1990-01-01

Programs that are intended to inform and provide "hands-on" experience for students and to facilitate the introduction of cell culture-based laboratory exercises into the high school and college laboratory are examined. The components of the CellServ Program and the Cell Culture Toxicology Training Programs are described. (KR)

Production of Trametes pubescens laccase under submerged and semi-solid culture conditions on agro-industrial wastes.

PubMed

Gonzalez, Juan C; Medina, Sandra C; Rodriguez, Alexander; Osma, Johann F; Alméciga-Díaz, Carlos J; Sánchez, Oscar F

2013-01-01

Laccases are copper-containing enzymes involved in the degradation of lignocellulosic materials and used in the treatment of phenol-containing wastewater. In this study we investigated the effect of culture conditions, i.e. submerged or semi-solid, and copper supplementation on laccase production by Trametespubescens grown on coffee husk, soybean pod husk, or cedar sawdust. The highest specific laccase activity was achieved when the culture was conducted under submerged conditions supplemented with copper (5 mM), and using coffee husk as substrate. The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1) and 60 kDa (Lac2). Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3. In addition, they showed similar stability and electro-chemical properties. At optimal culture conditions laccase activity was 7.69 ± 0.28 U mg(-1) of protein for the crude extract, and 0.08 ± 0.001 and 2.86 ± 0.05 U mg(-1) of protein for Lac1 and Lac2, respectively. In summary, these results show the potential of coffee husk as an important and economical growth medium to produce laccase, offering a new alternative use for this common agro-industrial byproduct.

Production of Trametes pubescens Laccase under Submerged and Semi-Solid Culture Conditions on Agro-Industrial Wastes

PubMed Central

Rodriguez, Alexander; Osma, Johann F.; Alméciga-Díaz, Carlos J.; Sánchez, Oscar F.

2013-01-01

Laccases are copper-containing enzymes involved in the degradation of lignocellulosic materials and used in the treatment of phenol-containing wastewater. In this study we investigated the effect of culture conditions, i.e. submerged or semi-solid, and copper supplementation on laccase production by Trametes pubescens grown on coffee husk, soybean pod husk, or cedar sawdust. The highest specific laccase activity was achieved when the culture was conducted under submerged conditions supplemented with copper (5 mM), and using coffee husk as substrate. The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1) and 60 kDa (Lac2). Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3. In addition, they showed similar stability and electro-chemical properties. At optimal culture conditions laccase activity was 7.69±0.28 U mg-1 of protein for the crude extract, and 0.08±0.001 and 2.86±0.05 U mg-1 of protein for Lac1 and Lac2, respectively. In summary, these results show the potential of coffee husk as an important and economical growth medium to produce laccase, offering a new alternative use for this common agro-industrial byproduct. PMID:24019936

Particle Size Distribution of Serratia marcescens Aerosols Created During Common Laboratory Procedures and Simulated Laboratory Accidents

PubMed Central

Kenny, Michael T.; Sabel, Fred L.

1968-01-01

Andersen air samplers were used to determine the particle size distribution of Serratia marcescens aerosols created during several common laboratory procedures and simulated laboratory accidents. Over 1,600 viable particles per cubic foot of air sampled were aerosolized during blending operations. More than 98% of these particles were less than 5 μ in size. In contrast, 80% of the viable particles aerosolized by handling lyophilized cultures were larger than 5 μ. Harvesting infected eggs, sonic treatment, centrifugation, mixing cultures, and dropping infectious material produced aerosols composed primarily of particles in the 1.0- to 7.5-μ size range. Images Fig. 1 PMID:4877498

Idaho National Laboratory Cultural Resource Monitoring Report for FY 2009

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

Brenda R. Pace; Julie B. Braun

2009-10-01

This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during fiscal year 2009 (FY 2009). Throughout the year, thirty-eight cultural resource localities were revisited including: two locations with Native American human remains, one of which is a cave, two additional caves, twenty-two prehistoric archaeological sites, six historic homesteads, two historic stage stations, two historic trails, and two nuclear resources, including Experimental Breeder Reactor-I, which is a designated National Historic Landmark. Several INL project areas were also monitored in FY 2009 to assess project compliance with cultural resource recommendations and monitormore » the effects of ongoing project activities. Although impacts were documented at a few locations and trespassing citations were issued in one instance, no significant adverse effects that would threaten the National Register eligibility of any resources were observed. Monitoring also demonstrated that several INL projects generally remain in compliance with recommendations to protect cultural resources.« less

Living Conditions and Psychological Distress in Latino Migrant Day Laborers: The Role of Cultural and Community Protective Factors.

PubMed

Organista, Kurt C; Ngo, Samantha; Neilands, Torsten B; Kral, Alex H

2017-03-01

The purpose of this study was to examine the relationship between typically difficult living conditions and psychological distress in Latino migrant day laborers (LMDLs), with attention to the potentially protective roles of contact with family in country of origin (i.e., communication, sending money, etc.), availability of local culture (i.e., food, music, people from one's country of origin), and utilization of community resources perceived to be culturally competent (i.e., services that are respectful, able to serve Latinos, able to solve problems, in Spanish, etc.). Participants were 344 LMDLs surveyed in the San Francisco Bay Area. As hypothesized: (a) difficult living conditions were related to depression, anxiety, and desesperación [desperation], the latter a popular Latino idiom of psychological distress recently validated on LMDLs; (b) contact with family moderated the relation between difficult living conditions and depression and desesperación but not anxiety and (c) access to local culture, and utilization of community resources, mediated the relation between difficult living conditions and depression and desesperación but not anxiety. Implications for intervening at local and larger levels in order to provide some protection against distress built into the LMDL experience in the United States are discussed. © Society for Community Research and Action 2016.

Oxygen Partial Pressure Is a Rate-Limiting Parameter for Cell Proliferation in 3D Spheroids Grown in Physioxic Culture Condition

PubMed Central

Gomes, Aurélie; Guillaume, Ludivine; Grimes, David Robert; Fehrenbach, Jérôme; Lobjois, Valérie; Ducommun, Bernard

2016-01-01

The in situ oxygen partial pressure in normal and tumor tissues is in the range of a few percent. Therefore, when studying cell growth in 3D culture systems, it is essential to consider how the physiological oxygen concentration, rather than the one in the ambient air, influences the proliferation parameters. Here, we investigated the effect of reducing oxygen partial pressure from 21% to 5% on cell proliferation rate and regionalization in a 3D tumor spheroid model. We found that 5% oxygen concentration strongly inhibited spheroid growth, changed the proliferation gradient and reduced the 50% In Depth Proliferation index (IDP50), compared with culture at 21% oxygen. We then modeled the oxygen partial pressure profiles using the experimental data generated by culturing spheroids in physioxic and normoxic conditions. Although hypoxia occurred at similar depth in spheroids grown in the two conditions, oxygen partial pressure was a major rate-limiting factor with a critical effect on cell proliferation rate and regionalization only in spheroids grown in physioxic condition and not in spheroids grown at atmospheric normoxia. Our findings strengthen the need to consider conducting experiment in physioxic conditions (i.e., tissue normoxia) for proper understanding of cancer cell biology and the evaluation of anticancer drugs in 3D culture systems. PMID:27575790

Oxygen Partial Pressure Is a Rate-Limiting Parameter for Cell Proliferation in 3D Spheroids Grown in Physioxic Culture Condition.

PubMed

Gomes, Aurélie; Guillaume, Ludivine; Grimes, David Robert; Fehrenbach, Jérôme; Lobjois, Valérie; Ducommun, Bernard

2016-01-01

The in situ oxygen partial pressure in normal and tumor tissues is in the range of a few percent. Therefore, when studying cell growth in 3D culture systems, it is essential to consider how the physiological oxygen concentration, rather than the one in the ambient air, influences the proliferation parameters. Here, we investigated the effect of reducing oxygen partial pressure from 21% to 5% on cell proliferation rate and regionalization in a 3D tumor spheroid model. We found that 5% oxygen concentration strongly inhibited spheroid growth, changed the proliferation gradient and reduced the 50% In Depth Proliferation index (IDP50), compared with culture at 21% oxygen. We then modeled the oxygen partial pressure profiles using the experimental data generated by culturing spheroids in physioxic and normoxic conditions. Although hypoxia occurred at similar depth in spheroids grown in the two conditions, oxygen partial pressure was a major rate-limiting factor with a critical effect on cell proliferation rate and regionalization only in spheroids grown in physioxic condition and not in spheroids grown at atmospheric normoxia. Our findings strengthen the need to consider conducting experiment in physioxic conditions (i.e., tissue normoxia) for proper understanding of cancer cell biology and the evaluation of anticancer drugs in 3D culture systems.

The antioxidant effect of the Malaysian Gelam honey on pancreatic hamster cells cultured under hyperglycemic conditions.

PubMed

Batumalaie, Kalaivani; Qvist, Rajes; Yusof, Kamaruddin Mohd; Ismail, Ikram Shah; Sekaran, Shamala Devi

2014-05-01

Type 2 diabetes consists of progressive hyperglycemia, insulin resistance, and pancreatic β-cell failure which could result from glucose toxicity, inflammatory cytokines, and oxidative stress. In the present study, we investigate the effect of pretreatment with Gelam honey (Melaleuca spp.) and the individual flavonoid components chrysin, luteolin, and quercetin, on the production of reactive oxygen species (ROS), cell viability, lipid peroxidation, and insulin content in hamster pancreatic cells (HIT-T15 cells), cultured under normal and hyperglycemic conditions. Phenolic extracts from a local Malaysian species of Gelam honey (Melaleuca spp.) were prepared using the standard extraction methods. HIT-T15 cells were cultured in 5 % CO2 and then preincubated with Gelam honey extracts (20, 40, 60, and 80 μg/ml) as well as some of its flavonoid components chrysin, luteolin, and quercetin (20, 40, 60, and 80 μM), prior to stimulation by 20 and 50 mM of glucose. The antioxidative effects were measured in these cultured cells at different concentrations and time point by DCFH-DA assay. Pretreatment of cells with Gelam honey extract or the flavonoid components prior to culturing in 20 or 50 mM glucose showed a significant decrease in the production of ROS, glucose-induced lipid peroxidation, and a significant increase in insulin content and the viability of cells cultured under hyperglycemic condition. Our results show the in vitro antioxidative property of the Gelam honey and the flavonoids on the β-cells from hamsters and its cytoprotective effect against hyperglycemia.

Cross-polarization microwave radar return at severe wind conditions: laboratory model and geophysical model function.

NASA Astrophysics Data System (ADS)

Troitskaya, Yuliya; Abramov, Victor; Ermoshkin, Alexey; Zuikova, Emma; Kazakov, Vassily; Sergeev, Daniil; Kandaurov, Alexandr

2014-05-01

Satellite remote sensing is one of the main techniques of monitoring severe weather conditions over the ocean. The principal difficulty of the existing algorithms of retrieving wind based on dependence of microwave backscattering cross-section on wind speed (Geophysical Model Function, GMF) is due to its saturation at winds exceeding 25 - 30 m/s. Recently analysis of dual- and quad-polarization C-band radar return measured from satellite Radarsat-2 suggested that the cross-polarized radar return has much higher sensitivity to the wind speed than co-polarized back scattering [1] and conserved sensitivity to wind speed at hurricane conditions [2]. Since complete collocation of these data was not possible and time difference in flight legs and SAR images acquisition was up to 3 hours, these two sets of data were compared in [2] only statistically. The main purpose of this paper is investigation of the functional dependence of cross-polarized radar cross-section on the wind speed in laboratory experiment. Since cross-polarized radar return is formed due to scattering at small-scale structures of the air-sea interface (short-crested waves, foam, sprays, etc), which are well reproduced in laboratory conditions, then the approach based on laboratory experiment on radar scattering of microwaves at the water surface under hurricane wind looks feasible. The experiments were performed in the Wind-wave flume located on top of the Large Thermostratified Tank of the Institute of Applied Physics, where the airflow was produced in the flume with the straight working part of 10 m and operating cross section 0.40?0.40 sq. m, the axis velocity can be varied from 5 to 25 m/s. Microwave measurements were carried out by a coherent Doppler X-band (3.2 cm) scatterometer with the consequent receive of linear polarizations. Experiments confirmed higher sensitivity to the wind speed of the cross-polarized radar return. Simultaneously parameters of the air flow in the turbulent boundary layer

Practical methods for handling human periodontal ligament stem cells in serum-free and serum-containing culture conditions under hypoxia: implications for regenerative medicine.

PubMed

Murabayashi, Dai; Mochizuki, Mai; Tamaki, Yuichi; Nakahara, Taka

2017-07-01

Stem cell-based therapies depend on the reliable expansion of patient-derived mesenchymal stem cells (MSCs) in vitro. The supplementation of cell culture media with serum is associated with several risks; accordingly, serum-free media are commercially available for cell culture. Furthermore, hypoxia is known to accelerate the expansion of MSCs. The present study aimed to characterize the properties of periodontal ligament-derived MSCs (PDLSCs) cultivated in serum-free and serum-containing media, under hypoxic and normoxic conditions. Cell growth, gene and protein expression, cytodifferentiation potential, genomic stability, cytotoxic response, and in vivo hard tissue generation of PDLSCs were examined. Our findings indicated that cultivation in serum-free medium does not affect the MSC phenotype or chromosomal stability of PDLSCs. PDLSCs expanded in serum-free medium exhibited more active growth than in fetal bovine serum-containing medium. We found that hypoxia does not alter the cell growth of PDLSCs under serum-free conditions, but inhibits their osteogenic and adipogenic cytodifferentiation while enabling maintenance of their multidifferentiation potential regardless of the presence of serum. PDLSCs expanded in serum-free medium were found to retain common MSC characteristics, including the capacity for hard tissue formation in vivo. However, PDLSCs cultured in serum-free culture conditions were more susceptible to damage following exposure to extrinsic cytotoxic stimuli than those cultured in medium supplemented with serum, suggesting that serum-free culture conditions do not exert protective effects against cytotoxicity on PDLSC cultures. The present work provides a comparative evaluation of cell culture in serum-free and serum-containing media, under hypoxic and normoxic conditions, for applications in regenerative medicine.

Effect of cultural conditions on antrodin C production by basidiomycete Antrodia camphorata in solid-state fermentation.

PubMed

Xia, Yongjun; Wang, Yuanlong; Zhang, Bobo; Xu, Ganrong; Ai, Lianzhong

2014-01-01

Antrodia camphorata is a medicinal fungus and antrodin C is one of the main bioactive components of A. camphorata in the submerged fermentation (SmF). To optimize the culture conditions, the factors influencing the production of antrodin C by A. camphorata under solid-state fermentation (SSF) were investigated in this study. Different solid substrates and external nitrogen sources were tested for their efficiency in producing antrodin C. The response surface methodology was applied to evaluate the influence of several variables, namely, the concentrations of soybean meal, initial moisture content, and inoculum density on antrodin C production in solid-state fermentation. The experimental results show that the optimum fermentation medium for antrodin C production by A. camphorata was composed of 0.578 g soybean meal, 0.05 g Na2 HPO4 , 0.05 g MgSO4 for 100 g rice, with 51.83% initial moisture content, 22 day culture time, 28 °C culture temperature, and 35.54% inoculum density. At optimized conditions, 6,617.36 ± 92.71 mg kg(-1) yield of antrodin C was achieved. Solid-state fermentation is one good cultural method to improve the production of antrodin C by A. camphorata. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

Demographics of cattle positive for Mycobacterium avium subspecies paratuberculosis by faecal culture, from submissions to the Cork Regional Veterinary Laboratory

PubMed Central

2009-01-01

The demography of bovine infections caused by Mycobacterium avium subspecies paratuberculosis (MAP) in Ireland is poorly defined. The objective of this study was to describe the demographics of cattle positive to MAP on faecal culture, based on submissions to the Cork Regional Veterinary Laboratory (Cork RVL) from 1994 to 2006. The study focused on all available faecal samples from adult cattle with non-responsive chronic diarrhoea that were submitted by private veterinary practitioners to Cork RVL for MAP culture. For each MAP-positive by faecal culture animal, data were collated from Cork RVL and Cattle Movement Monitoring Scheme (CMMS) records. Johne's disease (JD) was confirmed in 110 animals from 86 herds by the Cork RVL between 1994 and 2006, with a rate of positive cases between 15% and 18% over last four years of the study. Two breeds (Holstein/Friesian or Limousin) made up 78% of submissions. Movements were assessed for the 57 study animals with available movement information, 90% died within one year of the test and 26% tested positive in the herd they were born into. The study provides preliminary information about movement trends and demographics of animals with MAP positive submissions. Although the study area is restricted, it includes the most intensive (and economically-important) dairy region in Ireland. The demographics of JD infection from the study area are in agreement with international reports. Further work is required to determine demographic trends, incidence and prevalence of JD throughout Ireland. It is hoped this work may contribute to the development of a surveillance strategy for MAP by regional veterinary laboratories. PMID:21851736

Marine Biotoxins: Laboratory Culture and Molecular Structure

DTIC Science & Technology

1988-10-11

has been known for several years. has so far been isolated from toxic zoanthid corals. Palv~tha sp. Preliminary evidence suggests that the producer of...epiphyte of the zoanthid coral has not been realized. C. Purpose of Present Work The purposes of this work are the successful culture of G_. toxicus

Exploration Laboratory Analysis - ARC

NASA Technical Reports Server (NTRS)

Krihak, Michael K.; Fung, Paul P.

2012-01-01

The Exploration Laboratory Analysis (ELA) project supports the Exploration Medical Capability (ExMC) risk, Risk of Inability to Adequately Treat an Ill or Injured Crew Member, and ExMC Gap 4.05: Lack of minimally invasive in-flight laboratory capabilities with limited consumables required for diagnosing identified Exploration Medical Conditions. To mitigate this risk, the availability of inflight laboratory analysis instrumentation has been identified as an essential capability in future exploration missions. Mission architecture poses constraints on equipment and procedures that will be available to treat evidence-based medical conditions according to the Space Medicine Exploration Medical Conditions List (SMEMCL). The SMEMCL provided diagnosis and treatment for the evidence-based medical conditions and hence, a basis for developing ELA functional requirements.

In vitro storage of cedar shoot cultures under minimal growth conditions.

PubMed

Renau-Morata, Begoña; Arrillaga, Isabel; Segura, Juan

2006-07-01

We developed procedures for slow-growth storage of Cedrus atlantica and Cedrus libani microcuttings of juvenile and adult origin, noting factors favouring the extension of subculture intervals. Microcuttings could be stored effectively up to 6 months at 4 degrees C and reduced light intensity, provided that they were grown on a diluted modified MS medium. The addition of 6% mannitol to the storage media affected negatively survival and multiplication capacity of the cultures. The slow-growth storage conditions used in our experiments did not induce remarkable effects on both RAPD variability and average DNA methylation in the species.

FGF1 and IGF1-conditioned 3D culture system promoted the amplification and cancer stemness of lung cancer cells.

PubMed

Liu, Pengpeng; Zhang, Rui; Yu, Wenwen; Ye, Yingnan; Cheng, Yanan; Han, Lei; Dong, Li; Chen, Yongzi; Wei, Xiyin; Yu, Jinpu

2017-12-01

Lung cancer stem cells (LCSCs) are considered as the cellular origins of metastasis and relapse of lung cancer. However, routine two-dimensional culture system (2D-culture) hardly mimics the growth and functions of LCSCs in vivo and therefore significantly decreases the stemness activity of LCSCs. In this study, we constructed a special BME-based three-dimensional culture system (3D-culture) to amplify LCSCs in human lung adenocarcinoma cell line A549 cells and found 3D-culture promoted the enrichment and amplification of LCSCs in A549 cells displaying higher proliferation potential and invasion activity, but lower apoptosis. The expression and secretion levels of FGF1 and IGF1 were dramatically elevated in 3D-culture compared to 2D-culture. After growing in FGF1 and IGF1-conditioned 3D-culture, the proportion of LCSCs with specific stemness phenotypes in A549 cells significantly increased compared to that in conventional 3D suspension culture system. Further results indicated that FGF1 and IGF1 promoted the amplification and cancer stemness of LCSCs dependent on MAPK signaling pathway. Our data firstly established a growth factors-conditioned 3D-culture for LCSCs and demonstrated the effects of FGF1 and IGF1 in promoting the enrichment and amplification of LCSCs which might provide a feasible cell model in vitro for both mechanism study and translational research on lung cancer. Copyright © 2017 Elsevier Ltd. All rights reserved.

Synchronized mammalian cell culture: part I--a physical strategy for synchronized cultivation under physiological conditions.

PubMed

Barradas, Oscar Platas; Jandt, Uwe; Becker, Max; Bahnemann, Janina; Pörtner, Ralf; Zeng, An-Ping

2015-01-01

Conventional analysis and optimization procedures of mammalian cell culture processes mostly treat the culture as a homogeneous population. Hence, the focus is on cell physiology and metabolism, cell line development, and process control strategy. Impact on cultivations caused by potential variations in cellular properties between different subpopulations, however, has not yet been evaluated systematically. One main cause for the formation of such subpopulations is the progress of all cells through the cell cycle. The interaction of potential cell cycle specific variations in the cell behavior with large-scale process conditions can be optimally determined by means of (partially) synchronized cultivations, with subsequent population resolved model analysis. Therefore, it is desirable to synchronize a culture with minimal perturbation, which is possible with different yield and quality using physical selection methods, but not with frequently used chemical or whole-culture methods. Conventional nonsynchronizing methods with subsequent cell-specific, for example, flow cytometric analysis, can only resolve cell-limited effects of the cell cycle. In this work, we demonstrate countercurrent-flow centrifugal elutriation as a useful physical method to enrich mammalian cell populations within different phases of a cell cycle, which can be further cultivated for synchronized growth in bioreactors under physiological conditions. The presented combined approach contrasts with other physical selection methods especially with respect to the achievable yield, which makes it suitable for bioreactor scale cultivations. As shown with two industrial cell lines (CHO-K1 and human AGE1.HN), synchronous inocula can be obtained with overall synchrony degrees of up to 82% in the G1 phase, 53% in the S phase and 60% in the G2/M phase, with enrichment factors (Ysync) of 1.71, 1.79, and 4.24 respectively. Cells are able to grow with synchrony in bioreactors over several cell cycles. This

The impact of physiological oxygen during culture, and vitrification for cryopreservation, on the outcome of extended culture in human IVF.

PubMed

Gardner, David K

2016-02-01

Extended culture has facilitated the move to single blastocyst transfer, resulting in significant increases in implantation and live birth rate, while concomitantly reducing fetal loss during pregnancy. However, concerns have been raised regarding subsequent neo-natal outcomes following extended culture. Analysis of the literature reveals differences in outcomes according to geographical region and between individual clinics. A common factor amongst reports of potentially adverse outcomes following blastocyst transfer appears to be that atmospheric (~20%) oxygen was typically employed for embryo culture. Clinics and countries utilizing physiological concentrations of oxygen (~5%) have not reported adverse perinatal outcomes with blastocyst transfer. Atmospheric oxygen imposes significant negative effects upon the embryo's molecular and cellular physiology, and further it increases the sensitivity of the preimplantation embryo to other stressors in the laboratory. With the recent adoption of vitrification for blastocyst cryopreservation, cumulative pregnancy rates per cycle with extended culture will increase significantly. Consequently, rather than perceiving extended culture as a potentially negative procedure, it is concluded that neo-natal data need to be interpreted in light of the conditions used to culture and cryopreserve blastocysts, and that furthermore a policy of embryo culture using 20% oxygen can no longer be justified. Copyright © 2015 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Verification of the karst flow model under laboratory controlled conditions

NASA Astrophysics Data System (ADS)

Gotovac, Hrvoje; Andric, Ivo; Malenica, Luka; Srzic, Veljko

2016-04-01

Karst aquifers are very important groundwater resources around the world as well as in coastal part of Croatia. They consist of extremely complex structure defining by slow and laminar porous medium and small fissures and usually fast turbulent conduits/karst channels. Except simple lumped hydrological models that ignore high karst heterogeneity, full hydraulic (distributive) models have been developed exclusively by conventional finite element and finite volume elements considering complete karst heterogeneity structure that improves our understanding of complex processes in karst. Groundwater flow modeling in complex karst aquifers are faced by many difficulties such as a lack of heterogeneity knowledge (especially conduits), resolution of different spatial/temporal scales, connectivity between matrix and conduits, setting of appropriate boundary conditions and many others. Particular problem of karst flow modeling is verification of distributive models under real aquifer conditions due to lack of above-mentioned information. Therefore, we will show here possibility to verify karst flow models under the laboratory controlled conditions. Special 3-D karst flow model (5.6*2.6*2 m) consists of concrete construction, rainfall platform, 74 piezometers, 2 reservoirs and other supply equipment. Model is filled by fine sand (3-D porous matrix) and drainage plastic pipes (1-D conduits). This model enables knowledge of full heterogeneity structure including position of different sand layers as well as conduits location and geometry. Moreover, we know geometry of conduits perforation that enable analysis of interaction between matrix and conduits. In addition, pressure and precipitation distribution and discharge flow rates from both phases can be measured very accurately. These possibilities are not present in real sites what this model makes much more useful for karst flow modeling. Many experiments were performed under different controlled conditions such as different

Comparative bionomics of four populations of Meccus longipennis (Hemiptera: Reduviidae: Triatominae) under laboratory conditions.

PubMed

Martínez-Ibarra, José Alejandro; Nogueda-Torres, Benjamín; Licón-Trillo, Ángel; Villagrán-Herrera, María Elena; de Diego-Cabrera, José Antonio; Montañez-Valdez, Oziel Dante; Rocha-Chávez, Gonzalo

2013-04-01

The values of biological parameters related to the life cycles of four populations of Meccus longipennis (Reduviidae: Triatominae) were evaluated. Cohorts of each of the four studied populations from different geographical areas of Mexico were maintained under similar laboratory conditions and then compared. The population from El Saucito de Araujo was different from the other three studied populations, which could help explain the secondary importance of M. longipennis in the state of Chihuahua. This paper also supports the proposition that biological traits are important criteria for determining relationships between populations.

Roles of laboratories and laboratory systems in effective tuberculosis programmes.

PubMed

Ridderhof, John C; van Deun, Armand; Kam, Kai Man; Narayanan, P R; Aziz, Mohamed Abdul

2007-05-01

Laboratories and laboratory networks are a fundamental component of tuberculosis (TB) control, providing testing for diagnosis, surveillance and treatment monitoring at every level of the health-care system. New initiatives and resources to strengthen laboratory capacity and implement rapid and new diagnostic tests for TB will require recognition that laboratories are systems that require quality standards, appropriate human resources, and attention to safety in addition to supplies and equipment. To prepare the laboratory networks for new diagnostics and expanded capacity, we need to focus efforts on strengthening quality management systems (QMS) through additional resources for external quality assessment programmes for microscopy, culture, drug susceptibility testing (DST) and molecular diagnostics. QMS should also promote development of accreditation programmes to ensure adherence to standards to improve both the quality and credibility of the laboratory system within TB programmes. Corresponding attention must be given to addressing human resources at every level of the laboratory, with special consideration being given to new programmes for laboratory management and leadership skills. Strengthening laboratory networks will also involve setting up partnerships between TB programmes and those seeking to control other diseases in order to pool resources and to promote advocacy for quality standards, to develop strategies to integrate laboratories functions and to extend control programme activities to the private sector. Improving the laboratory system will assure that increased resources, in the form of supplies, equipment and facilities, will be invested in networks that are capable of providing effective testing to meet the goals of the Global Plan to Stop TB.

Animal serum-free culture conditions for isolation and expansion of multipotent mesenchymal stromal cells from human BM.

PubMed

Müller, I; Kordowich, S; Holzwarth, C; Spano, C; Isensee, G; Staiber, A; Viebahn, S; Gieseke, F; Langer, H; Gawaz, M P; Horwitz, E M; Conte, P; Handgretinger, R; Dominici, M

2006-01-01

Multipotent mesenchymal stromal cells (MSC) have become important tools in regenerative and transplantation medicine. Rapidly increasing numbers of patients are receiving in vitro-expanded MSC. Culture conditions typically include FSC because human serum does not fully support growth of human MSC in vitro (MSC(FCS)). Concerns regarding BSE, other infectious complications and host immune reactions have fueled investigation of alternative culture supplements. As PDGF has long been identified as a growth factor for MSC, we tested media supplementation with platelet lysate for support of MSC proliferation. We found that primary cultures of BM-derived MSC can be established with animal serum-free media containing fresh frozen plasma and platelets (MSC(FFPP)). Moreover, MSC(FFPP) showed vigorous proliferation that was superior to classical culture conditions containing FCS. MSC(FFPP) morphology was equivalent to MSC(FCS), and MSC(FFPP) expressed CD73, CD90, CD105, CD106, CD146 and HLA-ABC while being negative for CD34, CD45 and surface HLA-DR, as expected. In addition to being phenotypically identical, MSC(FFPP) could efficiently differentiate into adipocytes and osteoblasts. In terms of immune regulatory properties, MSC(FFPP) were indistinguishable from MSC(FCS). Proliferation of PBMC induced by IL-2 in combination with OKT-3 or by PHA was inhibited in the presence of MSC(FFPP). Taken together, FCS can be replaced safely by FFPP in cultures of MSC for clinical purposes.

Oxidative stress differentially impacts male and female bovine embryos depending on the culture medium and the stress condition.

PubMed

Dallemagne, Matthew; Ghys, Emmanuelle; De Schrevel, Catalina; Mwema, Ariane; De Troy, Delphine; Rasse, Catherine; Donnay, Isabelle

2018-09-01

Male and female embryos are known to differ for their metabolism and response to environmental factors very early in development. The present study aimed to evaluate the response to oxidative stress of male and female bovine embryos at the morula-blastocyst stages in terms of developmental rates, total cell number and apoptotic rates in two culture conditions. Embryos where cultured in a medium supplemented with either 5% fetal calf serum (FCS) or 4 mg/mL bovine serum albumin and a mixture of insulin, transferrin and selenium (BSA-ITS). Oxidative stress was applied at Day-5 post insemination (pi) by adding either AAPH or menadione to the culture medium, and blastocysts were analyzed at Day-7pi. The impact on development and blastocyst quality was dependent on the culture medium and the stress inducer but differed between male and female embryos. Male embryos resisted better to oxidative stress in FCS supplemented medium, no matter the stress inducer. Accordingly, the impact on blastocyst cell number tended to be higher in female blastocysts after stress induction with AAPH in FCS supplemented medium. On the other hand, in BSA-ITS supplemented medium, female embryos were more resistant to AAPH induced stress, while menadione had no impact on sex ratio. The weaker resistance of males to AAPH in this medium is in accordance with their trend to show a higher increase in apoptotic rates than females in this condition. In conclusion, this study shows that oxidative stress has differential impact on male and female bovine blastocysts depending on the culture condition and on the way oxidative stress is induced. Copyright © 2018 Elsevier Inc. All rights reserved.

Optimizing N-Fixing cyanobacteria culture to restore arid degraded soils

NASA Astrophysics Data System (ADS)

Roncero-Ramos, Beatriz; Román, Raúl; Gómez, Cintia; Chamizo, Sonia; Rodriguez-Caballero, Emilio; Cantón, Yolanda

2017-04-01

Cyanobacteria present several metabolic activities and mechanisms of adaptation which enable them to colonize different habitats, in almost all biome and continents, especially under extreme environmental conditions, as on the surface of the most arid soils and under the highest temperatures. In drylands, they are usually found among plants, cohabiting with organisms such as algae, lichens, mosses, bacteria and fungi, and in association with soil surface particles, forming communities known as biocrusts. Because they can survive under water stress and are considered ecosystem engineers, facilitating the establishment of other organisms, they can play a key role in the development of a successful restoration approach to recover the functionality of soils in arid and semiarid regions. In addition cyanobacteria can be cultured "ex-situ" obtaining high quantities of biomass to be used as soil inoculum at large scale. For these reasons, the inoculation of degrades soils with cyanobacteria can be considered an alternative to traditional restoration. This approach is expected to promote: the stabilization of the soil surface and the decrease of water and wind erosion; the increase of soil fertility by fixing N and C; and the succession of more developed organisms as mosses or vascular and annual plants. The objectives were: to evaluate the potential of a soil native cyanobacteria strain to be artificially cultured and the optimization of the process, and to analyze the effects of the inoculation of the biomass on soil under laboratory conditions. Cyanobacteria were isolated from biocrusts sampled on a limestone quarry located at the southeastern edge of the Sierra de Gádor massif (Spain). It was genetically and morphological identified as belonging to the nitrogen-fixing genera Nostoc. Essays were accomplished in bubble columns reactors (0.25 L), using different culture media: BG11+N, BG110, and two media made with fertilizers. Illumination simulated a circadian cycle

Nanoparticle growth and surface chemistry changes in cell-conditioned culture medium.

PubMed

Kendall, Michaela; Hodges, Nikolas J; Whitwell, Harry; Tyrrell, Jess; Cangul, Hakan

2015-02-05

When biomolecules attach to engineered nanoparticle (ENP) surfaces, they confer the particles with a new biological identity. Physical format may also radically alter, changing ENP stability and agglomeration state within seconds. In order to measure which biomolecules are associated with early ENP growth, we studied ENPs in conditioned medium from A549 cell culture, using dynamic light scattering (DLS) and linear trap quadrupole electron transfer dissociation mass spectrometry. Two types of 100 nm polystyrene particles (one uncoated and one with an amine functionalized surface) were used to measure the influence of surface type. In identically prepared conditioned medium, agglomeration was visible in all samples after 1 h, but was variable, indicating inter-sample variability in secretion rates and extracellular medium conditions. In samples conditioned for 1 h or more, ENP agglomeration rates varied significantly. Agglomerate size measured by DLS was well correlated with surface sequestered peptide number for uncoated but not for amine coated polystyrene ENPs. Amine-coated ENPs grew much faster and into larger agglomerates associated with fewer sequestered peptides, but including significant sequestered lactose dehydrogenase. We conclude that interference with extracellular peptide balance and oxidoreductase activity via sequestration is worthy of further study, as increased oxidative stress via this new mechanism may be important for cell toxicity. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Inquiring Scaffolds in Laboratory Tasks: An Instance of a "Worked Laboratory Guide Effect"?

ERIC Educational Resources Information Center

Schmidt-Borcherding, Florian; Hänze, Martin; Wodzinski, Rita; Rincke, Karsten

2013-01-01

The study explores if established support devices for paper-pencil problem solving, namely worked examples and incremental scaffolds, are applicable to laboratory tasks. N?=?173 grade eight students solved in dyads a physics laboratory task in one of three conditions. In condition A (unguided problem solving), students were asked to determine the…

Damage of natural stone tablets exposed to exhaust gas under laboratory conditions

NASA Astrophysics Data System (ADS)

Farkas, Orsolya; Szabados, György; Török, Ákos

2016-04-01

Natural stone tablets were exposed to exhaust gas under laboratory conditions to assess urban stone damage. Cylindrical test specimens (3 cm in diameter) were made from travertine, non-porous limestone, porous limestone, rhyolite tuff, sandstone, andesite, granite and marble. The samples were exposed to exhaust gas that was generated from diesel engine combustion (engine type: RÁBA D10 UTSLL 160, EURO II). The operating condition of the internal combustion engine was: 1300 r/m (app 50%). The exhaust gas was diverted into a pipe system where the samples were placed perpendicular to main flow for 1, 2, 4, 8 and 10 hours, respectively. The exhaust emission was measured by using AVL particulate measurement technology; filter paper method (AVL 415). The stone samples were documented and selective parameters were measured prior to and after exhaust gas exposure. Density, volume, ultrasonic pulse velocity, mineral composition and penetration depth of emission related particulate matter were recorded. The first results indicate that after 10 hours of exposure significant amount of particulate matter deposited on the stone surface independently from the surface properties and porosity. The black soot particles uniformly covered all types of stones, making hard to differentiate the specimens.

Intraspecific non-sexual interactions of Grammostola schulzei (Araneae: Theraphosidae) under laboratory conditions.

PubMed

Ferretti, Nelson E; Pérez-Miles, Fernando

2011-09-01

Intraspecific interactions of araneomorph spiders have received considerable attention, but there are few detailed studies on intraspecific interactions of mygalomorph spiders. Moreover, a thorough understanding of theraphosid biology and ecology is necessary from a conservation standpoint because natural populations may be threatened by habitat disturbances and captures for pet commerce. We described the behavior of conspecific individuals of Grammostola schulzei during non-sexual interactions, under laboratory conditions. Pairs of individuals involving adult males, adult females and juveniles were confronted and observed in resident and intruder conditions, totalizing 115 trials. When confronted two adult females, they retreated or grappled, and performed gaping display with bite attempts, usually resulted in severe injury of the intruder spiders. When confronted females with large juveniles, we frequently observed cannibalism on juveniles. Juveniles exposed to females or to other juveniles retreated or made leg tapping with forelegs and palpal drumming, which are common displays of courting adult males. Adult males courted and clasped some juveniles, but juveniles avoided or reject clasping. The behaviors observed during intraspecific interactions could play an important role determining spatial distribution and could lead to behavioral adaptations of territoriality.

Influence of culture conditions and medium composition on the production of antibacterial compounds by marine Serratia sp. WPRA3.

PubMed

Jafarzade, Mahtab; Yahya, Nur Ain; Shayesteh, Fatemeh; Usup, Gires; Ahmad, Asmat

2013-06-01

This study was undertaken to investigate the influence of culture conditions and medium components on production of antibacterial compounds by Serratia sp. WPRA3 (JX020764) which was isolated from marine water of Port Dickson, Malaysia. Biochemical, morphological, and molecular characteristics suggested that the isolate is a new candidate of the Serratia sp. The isolate showed strong antimicrobial activity against fungi, Gram-negative and Gram-positive bacteria. This bacterium exhibited optimum antibacterial compounds production at 28°C, pH 7 and 200 rev/min aeration during 72 h of incubation period. Highest antibacterial activity was obtained when sodium chloride (2%), yeast extract (0.5%), and glucose concentration (0.75%) were used as salt, nitrogen, and carbon sources respectively. Different active fractions were obtained by Thin-Layer Chromatography (TLC) and Flash Column Chromatography (FCC) from ethyl acetate crude extracts namely OCE and RCE in different culture conditions, OCE (pH 5, 200 rev/min) and RCE (pH 7/without aeration). In conclusion, the results suggested different culture conditions have a significant impact on the types of secondary metabolites produced by the bacterium.

Development of tissue culture techniques and hardware to study mineralization under microgravity conditions

NASA Astrophysics Data System (ADS)

van Loon, J. J. W. A.; Veldhuijzen, J. P.; Windgassen, E. J.; Brouwer, T.; Wattel, K.; van Vilsteren, M.; Maas, P.

1994-08-01

To study the effects of weightlessness on mouse fetal long bone rudiment growth and mineralization we have developed a tissue culture system for the Biorack facility of Spacelab. The technique uses standard liquid tissue culture medium, supplemented with Na-β-glycerophosphate, confined in gas permeable polyethylene bags mounted inside ESA Biorack Type I experiment containers. The containers can be flushed with an air/5% CO2 gas mixture necessary for the physiological bicarbonate buffer used. Small amounts of fluid can be introduced at the beginning (e.g. radioactive labels for incorporation studies) or at the end of the experiment (fixatives). A certain form of mechanical stimulation (continuous compression) can be used to counteract the, possibly, adverse effect of μ-gravity. Using 16 day old metatarsals the in vitro calcification process under μ-gravity conditions can be studied for a 4 day period.

Ruinous resident: the hydroid Ectopleura crocea negatively affects suspended culture of the mussel Mytilus galloprovincialis.

PubMed

Fitridge, Isla; Keough, Michael J

2013-01-01

Hydroids are major biofouling organisms in global aquaculture. Colonies of the hydroid Ectopleura crocea have recently established in Australian commercial mussel leases culturing Mytilus galloprovincialis. This study examined the impacts of E. crocea on mussel culture at two stages of the production cycle: spatfall and grow-out. Hydroids most commonly fouled the body, edge and dorsal regions of the mussel shell and cause a reduction in the length (4%) and weight (23%) of juvenile mussels. They also consumed mussel larvae in the field and in the laboratory. Prey numbers of many taxa, including mussel larvae, were consistent in natural hydroid diets regardless of the temporal variation in prey availability, implying some selectivity in hydroid feeding. In the laboratory, E. crocea consumed settling plantigrade mussel larvae more readily than trochophore or veliger larvae. Fouling by E. crocea is detrimental to mussel condition, and may affect the availability of wild mussel larvae in the commercial culture of M. galloprovincialis.

Roles of laboratories and laboratory systems in effective tuberculosis programmes

PubMed Central

van Deun, Armand; Kam, Kai Man; Narayanan, PR; Aziz, Mohamed Abdul

2007-01-01

Abstract Laboratories and laboratory networks are a fundamental component of tuberculosis (TB) control, providing testing for diagnosis, surveillance and treatment monitoring at every level of the health-care system. New initiatives and resources to strengthen laboratory capacity and implement rapid and new diagnostic tests for TB will require recognition that laboratories are systems that require quality standards, appropriate human resources, and attention to safety in addition to supplies and equipment. To prepare the laboratory networks for new diagnostics and expanded capacity, we need to focus efforts on strengthening quality management systems (QMS) through additional resources for external quality assessment programmes for microscopy, culture, drug susceptibility testing (DST) and molecular diagnostics. QMS should also promote development of accreditation programmes to ensure adherence to standards to improve both the quality and credibility of the laboratory system within TB programmes. Corresponding attention must be given to addressing human resources at every level of the laboratory, with special consideration being given to new programmes for laboratory management and leadership skills. Strengthening laboratory networks will also involve setting up partnerships between TB programmes and those seeking to control other diseases in order to pool resources and to promote advocacy for quality standards, to develop strategies to integrate laboratories’ functions and to extend control programme activities to the private sector. Improving the laboratory system will assure that increased resources, in the form of supplies, equipment and facilities, will be invested in networks that are capable of providing effective testing to meet the goals of the Global Plan to Stop TB. PMID:17639219

Studies of mineralization in tissue culture: optimal conditions for cartilage calcification

NASA Technical Reports Server (NTRS)

Boskey, A. L.; Stiner, D.; Doty, S. B.; Binderman, I.; Leboy, P.

1992-01-01

The optimal conditions for obtaining a calcified cartilage matrix approximating that which exists in situ were established in a differentiating chick limb bud mesenchymal cell culture system. Using cells from stage 21-24 embryos in a micro-mass culture, at an optimal density of 0.5 million cells/20 microliters spot, the deposition of small crystals of hydroxyapatite on a collagenous matrix and matrix vesicles was detected by day 21 using X-ray diffraction, FT-IR microscopy, and electron microscopy. Optimal media, containing 1.1 mM Ca, 4 mM P, 25 micrograms/ml vitamin C, 0.3 mg/ml glutamine, no Hepes buffer, and 10% fetal bovine serum, produced matrix resembling the calcifying cartilage matrix of fetal chick long bones. Interestingly, higher concentrations of fetal bovine serum had an inhibitory effect on calcification. The cartilage phenotype was confirmed based on the cellular expression of cartilage collagen and proteoglycan mRNAs, the presence of type II and type X collagen, and cartilage type proteoglycan at the light microscopic level, and the presence of chondrocytes and matrix vesicles at the EM level. The system is proposed as a model for evaluating the events in cell mediated cartilage calcification.

Degradation of trimethylbenzene isomers by an enrichment culture under N{sub 2}O-reducing conditions

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

Haener, A.; Hoehener, P.; Zeyer, J.

1997-03-01

In mineral oil-contaminated soils and aquifers, monoaromatic hydrocarbons are often a major concern because of high water solubility and toxicity. Under aerobic conditions, these compounds are rapidly mineralized. However, only limited data are available on the anaerobic degradation. This study reports on the growth of an enrichment culture on 1,3,5-trimethylbenzene (TMB) and 1,2,4-TMB under N2O-reducing conditions, and provides carbon mass and electron balances for the biodegradation of 1,3,5-TMB. 43 refs., 3 figs.

The laboratory diagnosis of syphilis.

PubMed

Ratnam, Sam

2005-01-01

Syphilis has several clinical manifestations, making laboratory testing a very important aspect of diagnosis. In North America, many unsuspected cases are discovered by laboratory testing. The etiological agent, Treponema pallidum, cannot be cultured, and there is no single optimal alternative test. Serological testing is the most frequently used approach in the laboratory diagnosis of syphilis. The present paper discusses the various serological and alternative tests currently available along with their limitations, and relates their results to the likely corresponding clinical stage of the disease. The need to use multiple tests is discussed, and the importance of quality control is noted. The complexity of syphilis serology means that the services of reference laboratories and clinical experts are often needed.

[The opportunity to use combined stem cells transplantation for haemopoesis activation in the old and mature laboratory animals under the conditions of ionizing radiation].

PubMed

Grebnev, D Iu; Maklakova, I Iu; Iastrebov, A P

2014-01-01

The objective of this work was to study the influence of combined transplantation of stem cells (multypotent mesenchimal stromal and haemopoetic stem cells) on the haemopoesis of old and mature laboratory animals under the condition of ionizing radiation. The result of the experiment shows that under physiological conditions the combined transplantation brings the erithropoesis activation, under the ionizing radiation conditions it brings the erythroid and granulocytopoesis activation. Moreover the combined MMSC and HSC transplantation gives cytoprotective action on the myeloid tissue due to decrease of cyto genically changed cells in the mature animals under the condition of ionizing radiation, but in the old animals this effect can be seen even under physiological condition. Combined transplantation of MMSC and GSC can be used in the mature and old laboratory animals under the conditions of ionising radiation for the haemopoesis activation.

Advanced cell culture techniques for cancer drug discovery.

PubMed

Lovitt, Carrie J; Shelper, Todd B; Avery, Vicky M

2014-05-30

Human cancer cell lines are an integral part of drug discovery practices. However, modeling the complexity of cancer utilizing these cell lines on standard plastic substrata, does not accurately represent the tumor microenvironment. Research into developing advanced tumor cell culture models in a three-dimensional (3D) architecture that more prescisely characterizes the disease state have been undertaken by a number of laboratories around the world. These 3D cell culture models are particularly beneficial for investigating mechanistic processes and drug resistance in tumor cells. In addition, a range of molecular mechanisms deconstructed by studying cancer cells in 3D models suggest that tumor cells cultured in two-dimensional monolayer conditions do not respond to cancer therapeutics/compounds in a similar manner. Recent studies have demonstrated the potential of utilizing 3D cell culture models in drug discovery programs; however, it is evident that further research is required for the development of more complex models that incorporate the majority of the cellular and physical properties of a tumor.

Advanced Cell Culture Techniques for Cancer Drug Discovery

PubMed Central

Lovitt, Carrie J.; Shelper, Todd B.; Avery, Vicky M.

2014-01-01

Human cancer cell lines are an integral part of drug discovery practices. However, modeling the complexity of cancer utilizing these cell lines on standard plastic substrata, does not accurately represent the tumor microenvironment. Research into developing advanced tumor cell culture models in a three-dimensional (3D) architecture that more prescisely characterizes the disease state have been undertaken by a number of laboratories around the world. These 3D cell culture models are particularly beneficial for investigating mechanistic processes and drug resistance in tumor cells. In addition, a range of molecular mechanisms deconstructed by studying cancer cells in 3D models suggest that tumor cells cultured in two-dimensional monolayer conditions do not respond to cancer therapeutics/compounds in a similar manner. Recent studies have demonstrated the potential of utilizing 3D cell culture models in drug discovery programs; however, it is evident that further research is required for the development of more complex models that incorporate the majority of the cellular and physical properties of a tumor. PMID:24887773

Survival, food consumption and growth of Norway lobster (Nephrops norvegicus) kept in laboratory conditions.

PubMed

Mente, Elena

2010-09-01

Successful commercial aquaculture of crustacean species is dependent on satisfying their nutritional requirements and on producing rapidly growing and healthy animals. The results of the present study provide valuable information for feeding habits and growth of Nephrops norvegicus L., 1758) under laboratory conditions. The aim of the present study was to examine food consumption, growth and physiology of the Norway lobster N. norvegicus under laboratory conditions. N. norvegicus (15 g wet weight) were distributed into 1001 tanks consisting of five numbered compartments each. They were fed the experimental diets (frozen mussels and pellets) for a period of 6 months. A group of starved Nephrops was stocked and fasted for 8 months. Although Nephrops grew well when fed the frozen mussels diet, feeding on a dry pellet feed was unsatisfactory. The starvation group, despite the fact that showed the highest mortality (50%), exhibited a remarkable tolerance to the lack of food supply. The study offers further insight by correlating the amino acid profiles of Nephrops tail muscle with the two diets. The deviations from the mussel's diet for asparagine, alanine and glutamic acid suggest a deficiency of these amino acids in this diet. The results of the present study showed that the concentrations of free amino acids are lower in relative amount than those of protein-bound amino acids, except for arginine, proline and glycine. The present study contributes to the improvement of our knowledge on nutritional requirements of the above species. © 2010 ISZS, Blackwell Publishing and IOZ/CAS.

Walk this way: validity evidence of iphone health application step count in laboratory and free-living conditions.

PubMed

Duncan, Markus J; Wunderlich, Kelly; Zhao, Yingying; Faulkner, Guy

2018-08-01

Several attempts have been made to demonstrate the accuracy of the iPhone pedometer function in laboratory test conditions. However, no studies have attempted to evaluate evidence of convergent validity of the iPhone step counts as a surveillance tool in the field. This study takes a pragmatic approach to evaluating Health application derived iPhone step counts by measuring accuracy of a standardized criterion iPhone SE and a heterogeneous sample of participant owned iPhones (6 or newer) in a laboratory condition, as well as comparing personal iPhones to accelerometer derived steps in a free-living test. During lab tests, criterion and personal iPhones differed from manually counted steps by a mean bias of less than ±5% when walking at 5km/h, 7.5km/h and 10km/h on a treadmill, which is generally considered acceptable for pedometers. In the free-living condition steps differed by a mean bias of 21.5% or 1340 steps/day when averaged across observation days. Researchers should be cautioned in considering the use of iPhone models as a research grade pedometer for physical activity surveillance or evaluation, likely due to the iPhone not being continually carried by participants; if compliance can be maximized then the iPhone might be suitable.

Anthocyanin production in callus cultures of Cleome rosea: modulation by culture conditions and characterization of pigments by means of HPLC-DAD/ESIMS.

PubMed

Simões, Claudia; Brasil Bizarri, Carlos Henrique; da Silva Cordeiro, Lívia; Carvalho de Castro, Tatiana; Machado Coutada, Leonardo César; Ribeiro da Silva, Antônio Jorge; Albarello, Norma; Mansur, Elisabeth

2009-10-01

Leaf and stem explants of Cleome rosea formed calluses when cultured on MS medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or 4-amino-3,5,6-trichloropicolinic acid (PIC). The highest biomass accumulation was obtained in the callus cultures initiated from stem explants on medium supplemented with 0.90 microM 2,4-D. Reddish-pink regions were observed on callus surface after 6-7 months in culture and these pigments were identified as anthocyanins. Anthocyanins production was enhanced by reducing temperature and increasing light irradiation. Pigmented calluses transferred to MS1/2 with a 1:4 ratio NH(4)(+)/NO(3)(-), 70 g L(-1) sucrose and supplementation with 0.90 microM 2,4-D maintained a high biomass accumulation and showed an increase of 150% on anthocyanin production as compared with the initial culture conditions. Qualitative analysis of calluses was performed by high performance liquid chromatography coupled to diode array detector and electrospray ionization mass spectrometry (HPLC-DAD/ESIMS). Eleven anthocyanins were characterized and the majority of them were identified as acylated cyanidins, although two peonidins were also detected. The major peak was composed by two anthocyanins, whose proposed identity were cyanidin 3-(p-coumaroyl) diglucoside-5-glucoside and cyanidin 3-(feruloyl) diglucoside-5-glucoside.

A 3D-psoriatic skin model for dermatological testing: The impact of culture conditions.

PubMed

Duque-Fernandez, Alexandra; Gauthier, Lydia; Simard, Mélissa; Jean, Jessica; Gendreau, Isabelle; Morin, Alexandre; Soucy, Jacques; Auger, Michèle; Pouliot, Roxane

2016-12-01

Inadequate representation of the human tissue environment during a preclinical screen can result in inaccurate predictions of compound effects. Consequently, pharmaceutical investigators are searching for preclinical models that closely resemble original tissue for predicting clinical outcomes. The current research aims to compare the impact of using serum-free medium instead of complete culture medium during the last step of psoriatic skin substitute reconstruction. Skin substitutes were produced according to the self-assembly approach. Serum-free conditions have no negative impact on the reconstruction of healthy or psoriatic skin substitutes presented in this study regarding their macroscopic or histological appearances. ATR-FTIR results showed no significant differences in the CH 2 bands between psoriatic substitutes cultured with or without serum, thus suggesting that serum deprivation did not have a negative impact on the lipid organization of their stratum corneum . Serum deprivation could even lead to a better organization of healthy skin substitute lipids. Percutaneous analyses demonstrated that psoriatic substitutes cultured in serum-free conditions showed a higher permeability to hydrocortisone compared to controls, while no significant differences in benzoic acid and caffeine penetration profiles were observed. Results obtained with this 3D-psoriatic skin substitute demonstrate the potential and versatility of the model. It could offer good prediction of drug related toxicities at preclinical stages performed in order to avoid unexpected and costly findings in the clinic. Together, these findings offer a new approach for one of the most important challenges of the 21st century, namely, prediction of drug toxicity.

Current and Past Strategies for Bacterial Culture in Clinical Microbiology

PubMed Central

Lagier, Jean-Christophe; Edouard, Sophie; Pagnier, Isabelle; Mediannikov, Oleg; Drancourt, Michel

2015-01-01

SUMMARY A pure bacterial culture remains essential for the study of its virulence, its antibiotic susceptibility, and its genome sequence in order to facilitate the understanding and treatment of caused diseases. The first culture conditions empirically varied incubation time, nutrients, atmosphere, and temperature; culture was then gradually abandoned in favor of molecular methods. The rebirth of culture in clinical microbiology was prompted by microbiologists specializing in intracellular bacteria. The shell vial procedure allowed the culture of new species of Rickettsia. The design of axenic media for growing fastidious bacteria such as Tropheryma whipplei and Coxiella burnetii and the ability of amoebal coculture to discover new bacteria constituted major advances. Strong efforts associating optimized culture media, detection methods, and a microaerophilic atmosphere allowed a dramatic decrease of the time of Mycobacterium tuberculosis culture. The use of a new versatile medium allowed an extension of the repertoire of archaea. Finally, to optimize the culture of anaerobes in routine bacteriology laboratories, the addition of antioxidants in culture media under an aerobic atmosphere allowed the growth of strictly anaerobic species. Nevertheless, among usual bacterial pathogens, the development of axenic media for the culture of Treponema pallidum or Mycobacterium leprae remains an important challenge that the patience and innovations of cultivators will enable them to overcome. PMID:25567228

Idaho National Laboratory Cultural Resource Monitoring Report for FY 2010

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

INL Cultural Resource Management Office

2010-10-01

This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during fiscal year 2010 (FY 2010). Throughout the year, thirty-three cultural resource localities were revisited, including somethat were visited more than once, including: two locations with Native American human remains, one of which is a cave, two additional caves, twenty-six prehistoric archaeological sites, two historic stage stations, and Experimental Breeder Reactor-I, which is a designated National Historic Landmark. The resources that were monitored included seventeen that are routinely visited and sixteen that are located in INL project areas. Although impacts weremore » documented at a few locations and one trespassing incident (albeit sans formal charges) was discovered, no significant adverse effects that would threaten the National Register eligibility of any resources were observed. Monitoring also demonstrated that several INL projects generally remain in compliance with recommendations to protect cultural resources.« less

Nutrient requirements and other factors involved in the culture of human kidney cells on microcarrier beads

NASA Technical Reports Server (NTRS)

Lewis, Marian L.; Morrison, Dennis R.

1987-01-01

The culture of human kidney cells on microcarrier beads in the Bioprocessing Laboratory at the Johnson Space Center is described. These were the first series of studies performed before and during 1983 to determine optimum conditions, including medium type, bead type and density. The composition of several medium types and the molecular weights of some common culture medium supplements and cellular proteins are included. The microgravity cell-to-bead attachment experiment performed on Space Transportation System Flight 8 is described.

Issues for laboratory outreach programs.

PubMed

1994-01-01

As we saw in the last "As We See It," many hospitals have begun outreach programs. We explored why outreach programs are established, the steps needed to develop a program, and the way to establish the proper business culture in a hospital laboratory for running a successful program. In this issue we identify the new skills laboratory managers need to be outreach managers, show how some programs maintain a competitive advantage, and explain some of the effects health-care reform will have on outreach services, as we ask: What are the requirements and issues involved in operating a successful laboratory outreach program?

INCIDENCE AND DETECTION OF PLEUROPNEUMONIA-LIKE ORGANISMS IN CELL CULTURES BY FLUORESCENT ANTIBODY AND CULTURAL PROCEDURES1

PubMed Central

Barile, Michael F.; Malizia, Walter F.; Riggs, Donald B.

1962-01-01

Barile, Michael F. (National Institutes of Health, Bethesda, Md.), Walter F. Malizia, and Donald B. Riggs. Incidence and detection of pleuropneumonia-like organisms in cell cultures by fluorescent antibody and cultural procedures. J. Bacteriol. 84:130–136. 1962—A total of 102 tissue-cell cultures from 17 separate laboratories was examined for pleuropneumonia-like organisms (PPLO) by the fluorescent antibody and cultural procedures. PPLO were isolated from 48 of the 49 tissue-cell cultures found positive for PPLO by the fluorescent antibody procedure, and results of the two procedures agreed in 101 of the 102 (99%) cases. PPLO were isolated from none of 10 primary-cell cultures prepared from six animal species and from 48 of 92 (52%) continuous-cell cultures prepared from eight animal species. Cells grown in media containing antibiotics were more frequently contaminated with PPLO (72%) than cells grown in antibiotic-free media (7%). Cultures (91%) from tissue-culture-producing laboratories and cultures (76%) used for propagation of microorganisms were contaminated with PPLO, although none used for tissue-culture metabolic studies was contaminated. In addition, our findings support the view that PPLO contamination of cell cultures is probably owing to bacterial contaminants which revert to L forms in the presence of antibiotics. Images PMID:13865001

Blood culture

MedlinePlus

... into (contaminating) the blood sample and causing a false-positive result (see below). The sample is sent to a laboratory. There, it is placed in a special dish (culture). It is then watched to see if bacteria ...

Cell Culture Made Easy.

ERIC Educational Resources Information Center

Dye, Frank J.

1985-01-01

Outlines steps to generate cell samples for observation and experimentation. The procedures (which use ordinary laboratory equipment) will establish a short-term primary culture of normal mammalian cells. Information on culture vessels and cell division and a list of questions to generate student interest and involvement in the topics are…

Simulation of outdoor pond cultures using indoor LED-lighted and temperature-controlled raceway ponds and Phenometrics photobioreactors

DOE PAGES

Huesemann, Michael; Dale, T.; Chavis, A.; ...

2016-12-02

Two innovative culturing systems, the LED-lighted and temperature-controlled 800 liter indoor raceways at Pacific Northwest National Laboratory (PNNL) and the Phenometrics environmental Photobioreactors™ (ePBRs) were evaluated in terms of their ability to accurately simulate the microalgae growth performance of outdoor cultures subjected to fluctuating sunlight and water temperature conditions. When repeating a 60-day outdoor pond culture experiment (batch and semi-continuous at two dilution rates) conducted in Arizona with the freshwater strain Chlorella sorokiniana DOE 1412 in these two indoor simulators, it was found that ash-free dry weight based biomass growth and productivity in the PNNL climate-simulation ponds was comparatively slightlymore » higher (8–13%) but significantly lower (44%) in the ePBRs. The difference in biomass productivities between the indoor and outdoor ponds was not statistically significant. When the marine Picochlorum soloecismus was cultured in five replicate ePBRs at Los Alamos National Laboratory (LANL) and in duplicate indoor climate-simulation ponds at PNNL, using the same inoculum, medium, culture depth, and light and temperature scripts, the optical density based biomass productivity and the rate of increase in cell counts in the ePBRs was about 35% and 66%, respectively, lower compared than in the indoor ponds. Potential reasons for the divergence in growth performance in these pond simulators, relative to outdoor raceways, are discussed. In conclusion, the PNNL climate-simulation ponds provide reasonably reliable biomass productivity estimates for microalgae strains cultured in outdoor raceways under different climatic conditions.« less

Sandia National Laboratories: Careers: Life at Sandia: People and Culture

Science.gov Websites

; Culture Work-Life Balance Special Programs Students and Postdocs Benefits and Perks Hiring Process Life at -Life Balance Careers People and Culture Solar cell picture Quality people, quality work Integrity , the incredible work-life balance." View All Jobs Equal opportunity employer/Disability/Vet/GLBT

Cultural Comparison of Chronic Conditions, Functional Status, and Acceptance in Older African-American and White Adults

PubMed Central

McDonald, Patricia E.; Zauszniewski, Jaclene A.; Bekhet, Abir K.

2010-01-01

Acceptance of functional decline accompanying chronic illness is challenging for all elders, and even more so for African-American elders. This study examined functional status and the number, types, and acceptance of chronic conditions in 16 African-American and 46 White elders. African-American elders reported better functioning but resembled Whites in number of chronic conditions and acceptance. All African-Americans reported hypertension; 76% of Whites reported arthritis. Greater acceptance was correlated with fewer chronic conditions (r = −.23, p < .05) and better functioning (r = −.59, p < .01). Poorer functioning (i.e., functional disability) was correlated with more chronic conditions (r = .27, p < .05). Culturally sensitive interventions are needed to enhance elders’ acceptance of chronic conditions and to improve their functioning. PMID:20857770

Effectiveness of practices to reduce blood culture contamination: A Laboratory Medicine Best Practices systematic review and meta-analysis☆

PubMed Central

Snyder, Susan R.; Favoretto, Alessandra M.; Baetz, Rich Ann; Derzon, James H.; Madison, Bereneice M.; Mass, Diana; Shaw, Colleen S.; Layfield, Christopher D.; Christenson, Robert H.; Liebow, Edward B.

2015-01-01

Objectives This article is a systematic review of the effectiveness of three practices for reducing blood culture contamination rates: venipuncture, phlebotomy teams, and prepackaged preparation/collection (prep) kits. Design and methods The CDC-funded Laboratory Medicine Best Practices Initiative systematic review methods for quality improvement practices were used. Results Studies included as evidence were: 9 venipuncture (vs. versus intravenous catheter), 5 phlebotomy team; and 7 prep kit. All studies for venipuncture and phlebotomy teams favored these practices, with meta-analysis mean odds ratios for venipuncture of 2.69 and phlebotomy teams of 2.58. For prep kits 6 studies’ effect sizes were not statistically significantly different from no effect (meta-analysis mean odds ratio 1.12). Conclusions Venipuncture and the use of phlebotomy teams are effective practices for reducing blood culture contamination rates in diverse hospital settings and are recommended as evidence-based “best practices” with high overall strength of evidence and substantial effect size ratings. No recommendation is made for or against prep kits based on uncertain improvement. PMID:22709932

Effectiveness of practices to reduce blood culture contamination: a Laboratory Medicine Best Practices systematic review and meta-analysis.

PubMed

Snyder, Susan R; Favoretto, Alessandra M; Baetz, Rich Ann; Derzon, James H; Madison, Bereneice M; Mass, Diana; Shaw, Colleen S; Layfield, Christopher D; Christenson, Robert H; Liebow, Edward B

2012-09-01

This article is a systematic review of the effectiveness of three practices for reducing blood culture contamination rates: venipuncture, phlebotomy teams, and prepackaged preparation/collection (prep) kits. The CDC-funded Laboratory Medicine Best Practices Initiative systematic review methods for quality improvement practices were used. Studies included as evidence were: 9 venipuncture (vs. versus intravenous catheter), 5 phlebotomy team; and 7 prep kit. All studies for venipuncture and phlebotomy teams favored these practices, with meta-analysis mean odds ratios for venipuncture of 2.69 and phlebotomy teams of 2.58. For prep kits 6 studies' effect sizes were not statistically significantly different from no effect (meta-analysis mean odds ratio 1.12). Venipuncture and the use of phlebotomy teams are effective practices for reducing blood culture contamination rates in diverse hospital settings and are recommended as evidence-based "best practices" with high overall strength of evidence and substantial effect size ratings. No recommendation is made for or against prep kits based on uncertain improvement. Copyright © 2012 The Canadian Society of Clinical Chemists. All rights reserved.

Growing Arabidopsis in vitro: cell suspensions, in vitro culture, and regeneration.

PubMed

Barkla, Bronwyn J; Vera-Estrella, Rosario; Pantoja, Omar

2014-01-01

An understanding of basic methods in Arabidopsis tissue culture is beneficial for any laboratory working on this model plant. Tissue culture refers to the aseptic growth of cells, organs, or plants in a controlled environment, in which physical, nutrient, and hormonal conditions can all be easily manipulated and monitored. The methodology facilitates the production of a large number of plants that are genetically identical over a relatively short growth period. Techniques, including callus production, cell suspension cultures, and plant regeneration, are all indispensable tools for the study of cellular biochemical and molecular processes. Plant regeneration is a key technology for successful stable plant transformation, while cell suspension cultures can be exploited for metabolite profiling and mining. In this chapter we report methods for the successful and highly efficient in vitro regeneration of plants and production of stable cell suspension lines from leaf explants of both Arabidopsis thaliana and Arabidopsis halleri.

A simple eccentric stirred tank mini-bioreactor: mixing characterization and mammalian cell culture experiments.

PubMed

Bulnes-Abundis, David; Carrillo-Cocom, Leydi M; Aráiz-Hernández, Diana; García-Ulloa, Alfonso; Granados-Pastor, Marisa; Sánchez-Arreola, Pamela B; Murugappan, Gayathree; Alvarez, Mario M

2013-04-01

In industrial practice, stirred tank bioreactors are the most common mammalian cell culture platform. However, research and screening protocols at the laboratory scale (i.e., 5-100 mL) rely primarily on Petri dishes, culture bottles, or Erlenmeyer flasks. There is a clear need for simple-easy to assemble, easy to use, easy to clean-cell culture mini-bioreactors for lab-scale and/or screening applications. Here, we study the mixing performance and culture adequacy of a 30 mL eccentric stirred tank mini-bioreactor. A detailed mixing characterization of the proposed bioreactor is presented. Laser induced fluorescence (LIF) experiments and computational fluid dynamics (CFD) computations are used to identify the operational conditions required for adequate mixing. Mammalian cell culture experiments were conducted with two different cell models. The specific growth rate and the maximum cell density of Chinese hamster ovary (CHO) cell cultures grown in the mini-bioreactor were comparable to those observed for 6-well culture plates, Erlenmeyer flasks, and 1 L fully instrumented bioreactors. Human hematopoietic stem cells were successfully expanded tenfold in suspension conditions using the eccentric mini-bioreactor system. Our results demonstrate good mixing performance and suggest the practicality and adequacy of the proposed mini-bioreactor. Copyright © 2012 Wiley Periodicals, Inc.

Exchange of nitrogen dioxide (NO2) between plants and the atmosphere under laboratory and field conditions

NASA Astrophysics Data System (ADS)

Breuninger, C.; Meixner, F. X.; Thielmann, A.; Kuhn, U.; Dindorf, T.; Kesselmeier, J.

2012-04-01

Nitric oxide (NO), nitrogen dioxide (NO2), often denoted as nitrogen oxides (NOx), and ozone (O3) are considered as most important compounds in atmospheric chemistry. In remote areas NOx concentration is related to biological activities of soils and vegetation. The emitted NOx will not entirely be subject of long range transport through the atmosphere. Aside oxidation of NO2 by the OH radical (forming HNO3), a considerable part of it is removed from the atmosphere through the uptake of NO2 by plants. The exchange depends on stomatal activity and on NO2 concentrations in ambient air. It is known that NO2 uptake by plants represents a large NO2 sink, but the magnitude and the NO2 compensation point concentration are still under discussion. Our dynamic chamber system allows exchange measurements of NO2 under field conditions (uncontrolled) as well as studies under controlled laboratory conditions including fumigation experiments. For NO2 detection we used a highly NO2 specific blue light converter (photolytic converter) with subsequent chemiluminescence analysis of the generated NO. Furthermore, as the exchange of NO2 is a complex interaction of transport, chemistry and plant physiology, in our field experiments we determined fluxes of NO, NO2, O3, CO2 and H2O. For a better knowledge of compensation point values for the bi-directional NO2 exchange we investigated a primary representative of conifers, Picea abies, under field and laboratory conditions, and re-analyzed older field data of the deciduous tree Quercus robur.

Optimization of cultural conditions for biosurfactant production by Pleurotus djamor in solid state fermentation.

PubMed

Velioglu, Zulfiye; Ozturk Urek, Raziye

2015-11-01

Being eco-friendly, less toxic, more biodegradable and biocompatible, biological surfactants have higher activity and stability compared to synthetic ones. In spite of the fact that there are abundant benefits of biosurfactants over the synthetic congeners, the problem related with the economical and large scale production proceeds. The utilization of several industrial wastes in the production media as substrates reduces the production cost. This current study aims optimization of biosurfactant production conditions by Pleurotus djamor, grown on sunflower seed shell, grape wastes or potato peels as renewable cheap substrates in solid state fermentation. After determination of the best substrate for biosurfactant production, we indicate optimum size and amount of solid substrate, volume of medium, temperature, pH and Fe(2+) concentrations on biosurfactant production. In optimum conditions, by reducing water surface tension to 28.82 ± 0.3 mN/m and having oil displacement diameter of 3.9 ± 0.3 cm, 10.205 ± 0.5 g/l biosurfactant was produced. Moreover, chemical composition of biosurfactant produced in optimum condition was determined by FTIR. Lastly, laboratory's large-scale production was carried out in optimum conditions in a tray bioreactor designed by us and 8.9 ± 0.5 g/l biosurfactant was produced with a significant surface activity (37.74 ± 0.3 mN/m). With its economical suggestions and applicability of laboratory's large-scale production, this work indicates the possibility of using low cost agro-industrial wastes as renewable substrates for biosurfactant production. Therefore, using economically produced biosurfactant will reduce cost in several applications such as bioremediation, oil recovery and biodegradation of toxic chemicals. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

INNOVATIONS IN EQUIPMENT AND TECHNIQUES FOR THE BIOLOGY TEACHING LABORATORY.

ERIC Educational Resources Information Center

BARTHELEMY, RICHARD E.; AND OTHERS

LABORATORY TECHNIQUES AND EQUIPMENT APPROPRIATE FOR TEACHING BIOLOGICAL SCIENCE CURRICULUM STUDY BIOLOGY ARE EMPHASIZED. MAJOR CATEGORIES INCLUDE (1) LABORATORY FACILITIES, (2) EQUIPMENT AND TECHNIQUES FOR CULTURE OF MICRO-ORGANISMS, (3) LABORATORY ANIMALS AND THEIR HOUSING, (4) TECHNIQUES FOR STUDYING PLANT GROWTH, (5) TECHNIQUES FOR STUDYING…

Development and Maturation of the Neuromuscular Junciton in Cell Culture Under Conditions of Simulated Zero-gravity

NASA Technical Reports Server (NTRS)

Gruener, R.

1985-01-01

Alterations in gravitational conditions which alter the normal development and interactions of nerve and muscle cells grown in culture is examined. Clinostat conditions, similating Og, which produce changes in cell morphology and growth patterns is studied. Data show that rotation of cocultures of nerve and muscle cells results in morphologic changes which are predicted to significantly alter the functional interactions between the elements of a prototypic synapse. It is further predicted that similar alterations may occur in central synapses which may therefore affect the development of the central nervous system when subjected to altered gravitational conditions.

Indoor cultivation and cultural characteristics of Wolfiporia cocos sclerotia using mushroom culture bottles.

PubMed

Kubo, Toshiyuki; Terabayashi, Susumu; Takeda, Shuichi; Sasaki, Hiroshi; Aburada, Masaki; Miyamoto, Ken-ichi

2006-06-01

We newly developed an indoor cultivation technique for Wolfiporia cocos (Wolf) Ryvarden et Gilbertson (Syn. Poria cocos Wolf), not with soil, but using mushroom culture bottles with pine logs, and clarified some cultural characteristics of sclerotia in the laboratory. To determine the optimum conditions for sclerotia growth, the weight of sclerotia and concentration of CO2 in three different air filters; cloth, paper and urethane resin, and closed bottles were tested. When the cloth air filter was used, the growth rate was the fastest and the yield was maximal. These results suggested that the aeration was an important environmental factor for cultivation. To clarify the characteristics of culture in the cloth air filtered and closed bottles, the weight of sclerotia, the compositions of pine logs and the contents of pachymic acid and dehydropachymic acid were examined during 24 weeks. The growth of scleroia and the wood decaying efficiency in the cloth air filtered bottles were better than those in the closed bottles. Also, it was found that W. cocos was a brown rot fungus due to the alkaline solubility of pine logs in the wood decay process. In addition, the contents of pachymic acid and dehydropachymic acid and the TLC pattern between the cultivated and commercial sclerotia did not differ remarkably.

Trends in Testing for Mycobacterium tuberculosis Complex From US Public Health Laboratories, 2009-2013.

PubMed

Tyrrell, Frances; Stafford, Cortney; Yakrus, Mitchell; Youngblood, Monica; Hill, Andrew; Johnston, Stephanie

We investigated data from US public health laboratories funded through the Centers for Disease Control and Prevention's Tuberculosis Elimination and Laboratory Cooperative Agreement to document trends and challenges in meeting national objectives in tuberculosis (TB) laboratory diagnoses. We examined data on workload and turnaround time from public health laboratories' progress reports during 2009-2013. We reviewed methodologies, laboratory roles, and progress toward rapid detection of Mycobacterium tuberculosis complex through nucleic acid amplification (NAA) testing. We compared selected data with TB surveillance reports to estimate public health laboratories' contribution to national diagnostic services. During the study period, culture and drug susceptibility tests decreased, but NAA testing increased. Public health laboratories achieved turnaround time benchmarks for drug susceptibility tests at lower levels than for acid-fast bacilli smear and identification from culture. NAA positivity in laboratories among surveillance-reported culture-positive TB cases increased from 26.6% (2355 of 8876) in 2009 to 40.0% (2948 of 7358) in 2013. Public health laboratories provided an estimated 50.9% (4285 of 8413 in 2010) to 57.2% (4210 of 7358 in 2013) of culture testing and 88.3% (6822 of 7727 in 2011) to 94.4% (6845 of 7250 in 2012) of drug susceptibility tests for all US TB cases. Public health laboratories contribute substantially to TB diagnoses in the United States. Although testing volumes mostly decreased, the increase in NAA testing indicates continued progress in rapid M tuberculosis complex detection.

SEE locomotor behavior test discriminates C57BL/6J and DBA/2J mouse inbred strains across laboratories and protocol conditions.

PubMed

Kafkafi, Neri; Lipkind, Dina; Benjamini, Yoav; Mayo, Cheryl L; Elmer, Gregory I; Golani, Ilan

2003-06-01

Conventional tests of behavioral phenotyping frequently have difficulties differentiating certain genotypes and replicating these differences across laboratories and protocol conditions. This study explores the hypothesis that automated tests can be designed to quantify ethologically relevant behavior patterns that more readily characterize heritable and replicable phenotypes. It used SEE (Strategy for the Exploration of Exploration) to phenotype the locomotor behavior of the C57BL/6 and DBA/2 mouse inbred strains across 3 laboratories. The 2 genotypes differed in 15 different measures of behavior, none of which had a significant genotype-laboratory interaction. Within the same laboratory, most of these differences were replicated in additional experiments despite the test photoperiod phase being changed and saline being injected. Results suggest that well-designed tests may considerably enhance replicability across laboratories.

Automation in the clinical microbiology laboratory.

PubMed

Novak, Susan M; Marlowe, Elizabeth M

2013-09-01

Imagine a clinical microbiology laboratory where a patient's specimens are placed on a conveyor belt and sent on an automation line for processing and plating. Technologists need only log onto a computer to visualize the images of a culture and send to a mass spectrometer for identification. Once a pathogen is identified, the system knows to send the colony for susceptibility testing. This is the future of the clinical microbiology laboratory. This article outlines the operational and staffing challenges facing clinical microbiology laboratories and the evolution of automation that is shaping the way laboratory medicine will be practiced in the future. Copyright © 2013 Elsevier Inc. All rights reserved.

Optimization of Cell Adhesion on Mg Based Implant Materials by Pre-Incubation under Cell Culture Conditions

PubMed Central

Willumeit, Regine; Möhring, Anneke; Feyerabend, Frank

2014-01-01

Magnesium based implants could revolutionize applications where orthopedic implants such as nails, screws or bone plates are used because they are load bearing and degrade over time. This prevents a second surgery to remove conventional implants. To improve the biocompatibility we studied here if and for how long a pre-incubation of the material under cell culture conditions is favorable for cell attachment and proliferation. For two materials, Mg and Mg10Gd1Nd, we could show that 6 h pre-incubation are already enough to form a natural protective layer suitable for cell culture. PMID:24857908

Optimization of cell adhesion on mg based implant materials by pre-incubation under cell culture conditions.

PubMed

Willumeit, Regine; Möhring, Anneke; Feyerabend, Frank

2014-05-05

Magnesium based implants could revolutionize applications where orthopedic implants such as nails, screws or bone plates are used because they are load bearing and degrade over time. This prevents a second surgery to remove conventional implants. To improve the biocompatibility we studied here if and for how long a pre-incubation of the material under cell culture conditions is favorable for cell attachment and proliferation. For two materials, Mg and Mg10Gd1Nd, we could show that 6 h pre-incubation are already enough to form a natural protective layer suitable for cell culture.

Laboratory Capacity for Antimicrobial Susceptibility Surveillance of Neisseria gonorrhoeae-District of Columbia, 2007-2012.

PubMed

Garrett, Tiana A; Davies-Cole, John; Furness, Bruce

2015-08-01

In the District of Columbia (DC), Neisseria gonorrhoeae (gonorrhea) infections accounted for more than 25% of 9321 incident sexually transmitted infections reported in 2011; untreated infections can lead to reproductive complications and a higher risk for HIV transmission. In DC, limited capacity to measure the prevalence of antibiotic-resistant N. gonorrhoeae is available; culture-based antibiotic susceptibility testing (AST) is needed to monitor antimicrobial resistance. We examined the capacity of laboratories that report to the DC Department of Health to perform AST for ongoing surveillance of antibiotic-resistant N. gonorrhoeae and to identify suspected treatment failures. We created a survey about diagnostic methods for gonorrhea testing and identified 33 laboratories that reported gonorrhea results to Department of Health in 2007 to 2012. Laboratories were assessed for use of bacterial culture or nucleic acid amplification testing (NAAT) for gonorrhea testing, prevalence of AST on gonorrhea-positive cultures, and types of antibiotics tested during AST. We estimated the prevalence of laboratory practices on the basis of self-report by staff. Nineteen (58%) laboratories completed the survey, representing 92% of the gonorrhea reporting. Seventeen (89%) of 19 laboratories conducted testing by culture; only 6 (35%) performed AST; 79% performed NAAT. Barriers to AST included longer completion times and limited number of provider requests for AST. Commercial laboratories (32%) were more likely to conduct both culture and NAAT, compared with health care facilities (11%). We report a low prevalence of laboratories performing AST because of multiple barriers. State-specific strategies addressing these barriers are needed to improve detection of antibiotic-resistant gonorrhea stains circulating among the population.

Atrazine degradation by fungal co-culture enzyme extracts under different soil conditions.

PubMed

Chan-Cupul, Wilberth; Heredia-Abarca, Gabriela; Rodríguez-Vázquez, Refugio

2016-01-01

This investigation was undertaken to determine the atrazine degradation by fungal enzyme extracts (FEEs) in a clay-loam soil microcosm contaminated at field application rate (5 μg g(-1)) and to study the influence of different soil microcosm conditions, including the effect of soil sterilization, water holding capacity, soil pH and type of FEEs used in atrazine degradation through a 2(4) factorial experimental design. The Trametes maxima-Paecilomyces carneus co-culture extract contained more laccase activity and hydrogen peroxide (H2O2) content (laccase = 18956.0 U mg protein(-1), H2O2 = 6.2 mg L(-1)) than the T. maxima monoculture extract (laccase = 12866.7 U mg protein(-1), H2O2 = 4.0 mg L(-1)). Both extracts were able to degrade atrazine at 100%; however, the T. maxima monoculture extract (0.32 h) achieved a lower half-degradation time than its co-culture with P. carneus (1.2 h). The FEE type (p = 0.03) and soil pH (p = 0.01) significantly affected atrazine degradation. The best degradation rate was achieved by the T. maxima monoculture extract in an acid soil (pH = 4.86). This study demonstrated that both the monoculture extracts of the native strain T. maxima and its co-culture with P. carneus can efficiently and quickly degrade atrazine in clay-loam soils.

Definition of culture conditions for Arxula adeninivorans, a rational basis for studying heterologous gene expression in this dimorphic yeast.

PubMed

Stöckmann, Christoph; Palmen, Thomas G; Schroer, Kirsten; Kunze, Gotthard; Gellissen, Gerd; Büchs, Jochen

2014-06-01

The yeast Arxula adeninivorans is considered to be a promising producer of recombinant proteins. However, growth characteristics are poorly investigated and no industrial process has been established yet. Though of vital interest for strain screening and production processes, rationally defined culture conditions remain to be developed. A cultivation system was evolved based on targeted sampling and mathematical analysis of rationally designed small-scale cultivations in shake flasks. The oxygen and carbon dioxide transfer rates were analyzed as conclusive online parameters. Oxygen limitation extended cultivation and led to ethanol formation in cultures supplied with glucose. Cultures were inhibited at pH-values below 2.8. The phosphorus demand was determined as 1.55 g phosphorus per 100 g cell dry weight. Synthetic SYN6 medium with 20 g glucose l(-1) was optimized for cultivation in shake flasks by buffering at pH 6.4 with 140 mmol MES l(-1). Optimized SYN6 medium and operating conditions provided non-limited cultivations without by-product formation. A maximal specific growth rate of 0.32 h(-1) and short fermentations of 15 h were achieved. A pH optimum curve was derived from the oxygen transfer rates of differently buffered cultures, showing maximal growth between pH 2.8 and 6.5. Furthermore, it was shown that the applied medium and cultivation conditions were also suitable for non-limiting growth and product formation of a genetically modified A. adeninivorans strain expressing a heterologous phytase.

Adherence of Escherichia coli O157:H7 to epithelial cells in vitro and in pig gut loops is affected by bacterial culture conditions

PubMed Central

Yin, Xianhua; Feng, Yanni; Wheatcroft, Roger; Chambers, James; Gong, Joshua; Gyles, Carlton L.

2011-01-01

The objectives of this study were to determine the effect of bacterial culture conditions on adherence of enterohemorrhagic Escherichia coli (EHEC) O157:H7 strain 86-24 in vivo to pig enterocytes and to compare the results with adherence in vitro to cultured HEp-2 and IPEC-J2 cells. Growth of O157:H7 in MacConkey broth (MB) resulted in almost no adherence to both HEp-2 and IPEC-J2 cells; prior exposure of the bacteria to pH 2.5 reduced adherence. There was greater adherence by bacteria from static cultures than by those from shaken cultures and by bacteria cultured in brain–heart infusion (BHI) plus NaHCO3 (BHIN) than by bacteria cultured in BHI. In contrast, in pig ileal loops, bacteria cultured in MB adhered well to enterocytes, and prior exposure to pH 2.5 had no effect on adherence. Among several media tested for their effect on bacterial adherence in the pig intestine, MB and BHIN proved to be the best. Bacterial adherence was dose-dependent and was more extensive in the ileum than in the colon. This study demonstrated that there are remarkable differences between culture conditions that promote adherence of an EHEC O157:H7 strain in vitro and in vivo, that culture conditions profoundly affect adherence to epithelial cells in vitro and in vivo, and that pig ileal loops are better suited to adherence studies than are colon loops. PMID:21731177

Laboratory mouse housing conditions can be improved using common environmental enrichment without compromising data.

PubMed

André, Viola; Gau, Christine; Scheideler, Angelika; Aguilar-Pimentel, Juan A; Amarie, Oana V; Becker, Lore; Garrett, Lillian; Hans, Wolfgang; Hölter, Sabine M; Janik, Dirk; Moreth, Kristin; Neff, Frauke; Östereicher, Manuela; Racz, Ildiko; Rathkolb, Birgit; Rozman, Jan; Bekeredjian, Raffi; Graw, Jochen; Klingenspor, Martin; Klopstock, Thomas; Ollert, Markus; Schmidt-Weber, Carsten; Wolf, Eckhard; Wurst, Wolfgang; Gailus-Durner, Valérie; Brielmeier, Markus; Fuchs, Helmut; Hrabé de Angelis, Martin

2018-04-01

Animal welfare requires the adequate housing of animals to ensure health and well-being. The application of environmental enrichment is a way to improve the well-being of laboratory animals. However, it is important to know whether these enrichment items can be incorporated in experimental mouse husbandry without creating a divide between past and future experimental results. Previous small-scale studies have been inconsistent throughout the literature, and it is not yet completely understood whether and how enrichment might endanger comparability of results of scientific experiments. Here, we measured the effect on means and variability of 164 physiological parameters in 3 conditions: with nesting material with or without a shelter, comparing these 2 conditions to a "barren" regime without any enrichments. We studied a total of 360 mice from each of 2 mouse strains (C57BL/6NTac and DBA/2NCrl) and both sexes for each of the 3 conditions. Our study indicates that enrichment affects the mean values of some of the 164 parameters with no consistent effects on variability. However, the influence of enrichment appears negligible compared to the effects of other influencing factors. Therefore, nesting material and shelters may be used to improve animal welfare without impairment of experimental outcome or loss of comparability to previous data collected under barren housing conditions.

Laboratory mouse housing conditions can be improved using common environmental enrichment without compromising data

PubMed Central

Gau, Christine; Scheideler, Angelika; Aguilar-Pimentel, Juan A.; Amarie, Oana V.; Becker, Lore; Garrett, Lillian; Hans, Wolfgang; Hölter, Sabine M.; Janik, Dirk; Moreth, Kristin; Neff, Frauke; Östereicher, Manuela; Racz, Ildiko; Rathkolb, Birgit; Rozman, Jan; Bekeredjian, Raffi; Graw, Jochen; Klingenspor, Martin; Klopstock, Thomas; Ollert, Markus; Schmidt-Weber, Carsten; Wolf, Eckhard; Wurst, Wolfgang; Gailus-Durner, Valérie; Brielmeier, Markus; Fuchs, Helmut; Hrabé de Angelis, Martin

2018-01-01

Animal welfare requires the adequate housing of animals to ensure health and well-being. The application of environmental enrichment is a way to improve the well-being of laboratory animals. However, it is important to know whether these enrichment items can be incorporated in experimental mouse husbandry without creating a divide between past and future experimental results. Previous small-scale studies have been inconsistent throughout the literature, and it is not yet completely understood whether and how enrichment might endanger comparability of results of scientific experiments. Here, we measured the effect on means and variability of 164 physiological parameters in 3 conditions: with nesting material with or without a shelter, comparing these 2 conditions to a “barren” regime without any enrichments. We studied a total of 360 mice from each of 2 mouse strains (C57BL/6NTac and DBA/2NCrl) and both sexes for each of the 3 conditions. Our study indicates that enrichment affects the mean values of some of the 164 parameters with no consistent effects on variability. However, the influence of enrichment appears negligible compared to the effects of other influencing factors. Therefore, nesting material and shelters may be used to improve animal welfare without impairment of experimental outcome or loss of comparability to previous data collected under barren housing conditions. PMID:29659570

Insights into an Optimization of Plasmodium vivax Sal-1 In Vitro Culture: The Aotus Primate Model.

PubMed

Shaw-Saliba, Kathryn; Thomson-Luque, Richard; Obaldía, Nicanor; Nuñez, Marlon; Dutary, Sahir; Lim, Caeul; Barnes, Samantha; Kocken, Clemens H M; Duraisingh, Manoj T; Adams, John H; Pasini, Erica M

2016-07-01

Malaria is one of the most significant tropical diseases, and of the Plasmodium species that cause human malaria, P. vivax is the most geographically widespread. However, P. vivax remains a relatively neglected human parasite since research is typically limited to laboratories with direct access to parasite isolates from endemic field settings or from non-human primate models. This restricted research capacity is in large part due to the lack of a continuous P. vivax in vitro culture system, which has hampered the ability for experimental research needed to gain biological knowledge and develop new therapies. Consequently, efforts to establish a long-term P. vivax culture system are confounded by our poor knowledge of the preferred host cell and essential nutrients needed for in vitro propagation. Reliance on very heterogeneous P. vivax field isolates makes it difficult to benchmark parasite characteristics and further complicates development of a robust and reliable culture method. In an effort to eliminate parasite variability as a complication, we used a well-defined Aotus-adapted P. vivax Sal-1 strain to empirically evaluate different short-term in vitro culture conditions and compare them with previous reported attempts at P. vivax in vitro culture Most importantly, we suggest that reticulocyte enrichment methods affect invasion efficiency and we identify stabilized forms of nutrients that appear beneficial for parasite growth, indicating that P. vivax may be extremely sensitive to waste products. Leuko-depletion methods did not significantly affect parasite development. Formatting changes such as shaking and static cultures did not seem to have a major impact while; in contrast, the starting haematocrit affected both parasite invasion and growth. These results support the continued use of Aotus-adapted Sal-1 for development of P. vivax laboratory methods; however, further experiments are needed to optimize culture conditions to support long-term parasite

Insights into an Optimization of Plasmodium vivax Sal-1 In Vitro Culture: The Aotus Primate Model

PubMed Central

Obaldía, Nicanor; Nuñez, Marlon; Dutary, Sahir; Lim, Caeul; Barnes, Samantha; Kocken, Clemens H. M.; Duraisingh, Manoj T.; Adams, John H.; Pasini, Erica M.

2016-01-01

Malaria is one of the most significant tropical diseases, and of the Plasmodium species that cause human malaria, P. vivax is the most geographically widespread. However, P. vivax remains a relatively neglected human parasite since research is typically limited to laboratories with direct access to parasite isolates from endemic field settings or from non-human primate models. This restricted research capacity is in large part due to the lack of a continuous P. vivax in vitro culture system, which has hampered the ability for experimental research needed to gain biological knowledge and develop new therapies. Consequently, efforts to establish a long-term P. vivax culture system are confounded by our poor knowledge of the preferred host cell and essential nutrients needed for in vitro propagation. Reliance on very heterogeneous P. vivax field isolates makes it difficult to benchmark parasite characteristics and further complicates development of a robust and reliable culture method. In an effort to eliminate parasite variability as a complication, we used a well-defined Aotus-adapted P. vivax Sal-1 strain to empirically evaluate different short-term in vitro culture conditions and compare them with previous reported attempts at P. vivax in vitro culture Most importantly, we suggest that reticulocyte enrichment methods affect invasion efficiency and we identify stabilized forms of nutrients that appear beneficial for parasite growth, indicating that P. vivax may be extremely sensitive to waste products. Leuko-depletion methods did not significantly affect parasite development. Formatting changes such as shaking and static cultures did not seem to have a major impact while; in contrast, the starting haematocrit affected both parasite invasion and growth. These results support the continued use of Aotus-adapted Sal-1 for development of P. vivax laboratory methods; however, further experiments are needed to optimize culture conditions to support long-term parasite

Culture conditions and salt effects on essential oil composition of sweet marjoram (Origanum majorana) from Tunisia.

PubMed

Baâtour, Olfa; Tarchoune, Imen; Mahmoudi, Hela; Nassri, Nawel; Abidi, Wissal; Kaddour, Rym; Hamdaoui, Ghaith; Nasri-Ayachi, Mouhiba Ben; Lachaâl, Mohtar; Marzouk, Brahim

2012-06-01

O. majorana shoots were investigated for their essential oil (EO) composition. Two experiments were carried out; the first on hydroponic medium in a culture chamber and the second on inert sand in a greenhouse for 20 days. Plants were cultivated for 17 days in hydroponic medium supplemented with NaCl 100 mmol L⁻¹. The results showed that the O. majorana hydroponic medium offered higher essential oil yield than that from the greenhouse. The latter increased significantly in yield (by 50 %) under saline constraint while it did not change in the culture chamber. Under greenhouse conditions and in the absence of salt treatment, the major constituents were terpinen-4-ol and trans-sabinene hydrate. However, in the culture chamber, the major volatile components were cis-sabinene hydrate and terpinen-4-ol. In the presence of NaCl, new compounds appeared, such as eicosane, spathulenol, eugenol, and phenol. In addition, in the greenhouse, with or without salt, a very important change of trans-sabinene hydrate concentration in EO occurred, whereas in the culture chamber change appeared in cis-sabinene hydrate content.

Physicochemical characterization of engineered nanoparticles under physiological conditions: effect of culture media components and particle surface coating.

PubMed

Fatisson, Julien; Quevedo, Ivan R; Wilkinson, Kevin J; Tufenkji, Nathalie

2012-03-01

The use of engineered nanoparticles (ENPs) in commercial products has increased substantially over the last few years. Some research has been conducted in order to determine whether or not such materials are cytotoxic, but questions remain regarding the role that physiological media and sera constituents play in ENP aggregation or stabilization. In this study, several characterization methods were used to evaluate the particle size and surface potential of 6 ENPs suspended in a number of culture media and in the presence of different culture media constituents. Dynamic light scattering (DLS) and fluorescence correlation spectroscopy (FCS) were employed for size determinations. Results were interpreted on the basis of ENP surface potentials evaluated from particle electrophoretic mobilities (EPM). Measurements made after 24h of incubation at 37°C showed that the cell culture medium constituents had only moderate impact on the physicochemical properties of the ENP, although incubation in bovine serum albumin destabilized the colloidal system. In contrast, most of the serum proteins increased colloidal stabilization. Moreover, the type of ENP surface modification played a significant role in ENP behavior whereby the complexity of interactions between the ENPs and the medium components generally decreased with increasing complexity of the particle surface. This investigation emphasizes the importance of ENP characterization under conditions that are representative of cell culture media or physiological conditions for improved assessments of nanoparticle cytotoxicity. Copyright © 2011 Elsevier B.V. All rights reserved.

Biotransformation of BTEX under anaerobic, denitrifying conditions: Field and laboratory observations

NASA Astrophysics Data System (ADS)

Barbaro, J. R.; Barker, J. F.; Lemon, L. A.; Mayfield, C. I.

1992-11-01

Three natural-gradient injection experiments in the Borden aquifer (Ontario, Canada) (˜ 100-300 days in duration) and a 452-day laboratory microcosm experiment were performed to evaluate the biotransformation of BTEX (benzene, toluene, ethylbenzene and o-, m-, p-xylenes) derived from gasoline under anaerobic, denitrifying conditions. Both NO 3-- amended and unamended control (i.e. no NO 3- added) experiments were performed. In the unamended control injection experiment, toluene biotransformed between 1 and 5 m from the injection well. All other aromatic compounds were recalcitrant in this field experiment and all aromatic compounds were recalcitrant in unamended control microcosms. After an acclimatization period, toluene biotransformed relatively rapidly in the presence of NO 3- in both the laboratory and field to a residual level of ˜ 100 μg L -1. In the presence of NO 3- the xylene isomers and ethylbenzene biotransformed to a lesser degree. Benzene was recalcitrant in all experiments. The acetylene blockage technique was used to demonstrate that denitrifying bacteria were active in the presence of NO 3-. In the NO 3--amended injection experiments, little BTEX mass loss occurred beyond the 1-m multilevel-piezometer fence. However, NO 3- continued to decline downgradient, suggesting that other sources of carbon were being utilized by denitrifying bacteria in preference to residual BTEX. In addition to observations on mass loss, these experiments provided evidence of inhibition of BTEX biotransformation in the presence of acetylene, and competitive utilization between toluene, ethylbenzene and the xylene isomers. Given the recalcitrance of benzene and high thresholds of the compounds that did biotransform, the addition of NO 3- as an alternate electron acceptor would not be successful in this aquifer as a remedial measure.

Do hypoxia/normoxia culturing conditions change the neuroregulatory profile of Wharton Jelly mesenchymal stem cell secretome?

PubMed

Teixeira, Fábio G; Panchalingam, Krishna M; Anjo, Sandra Isabel; Manadas, Bruno; Pereira, Ricardo; Sousa, Nuno; Salgado, António J; Behie, Leo A

2015-07-24

The use of human umbilical cord Wharton Jelly-derived mesenchymal stem cells (hWJ-MSCs) has been considered a new potential source for future safe applications in regenerative medicine. Indeed, the application of hWJ-MSCs into different animal models of disease, including those from the central nervous system, has shown remarkable therapeutic benefits mostly associated with their secretome. Conventionally, hWJ-MSCs are cultured and characterized under normoxic conditions (21 % oxygen tension), although the oxygen levels within tissues are typically much lower (hypoxic) than these standard culture conditions. Therefore, oxygen tension represents an important environmental factor that may affect the performance of mesenchymal stem cells in vivo. However, the impact of hypoxic conditions on distinct mesenchymal stem cell characteristics, such as the secretome, still remains unclear. In the present study, we have examined the effects of normoxic (21 % O2) and hypoxic (5 % O2) conditions on the hWJ-MSC secretome. Subsequently, we address the impact of the distinct secretome in the neuronal cell survival and differentiation of human neural progenitor cells. The present data indicate that the hWJ-MSC secretome collected from normoxic and hypoxic conditions displayed similar effects in supporting neuronal differentiation of human neural progenitor cells in vitro. However, proteomic analysis revealed that the use of hypoxic preconditioning led to the upregulation of several proteins within the hWJ-MSC secretome. Our results suggest that the optimization of parameters such as hypoxia may lead to the development of strategies that enhance the therapeutic effects of the secretome for future regenerative medicine studies and applications.

Assessment of the Laboratory Learning Environment in an Inquiry-Oriented Chemistry Laboratory in Arab and Jewish High Schools in Israel

ERIC Educational Resources Information Center

Dkeidek, Iyad; Mamlok-Naaman, Rachel; Hofstein, Avi

2012-01-01

An inquiry-oriented laboratory in chemistry was integrated into the chemistry curriculum in Jewish high schools in Israel, and after a short period was also implemented in Arab sector. In this study, we investigated the effect of culture on the perceptions of laboratory classroom learning environments by comparing the perceptions of Arab and…

Laboratory Diagnosis of Infective Endocarditis

PubMed Central

Liesman, Rachael M.; Pritt, Bobbi S.; Maleszewski, Joseph J.

2017-01-01

ABSTRACT Infective endocarditis is life-threatening; identification of the underlying etiology informs optimized individual patient management. Changing epidemiology, advances in blood culture techniques, and new diagnostics guide the application of laboratory testing for diagnosis of endocarditis. Blood cultures remain the standard test for microbial diagnosis, with directed serological testing (i.e., Q fever serology, Bartonella serology) in culture-negative cases. Histopathology and molecular diagnostics (e.g., 16S rRNA gene PCR/sequencing, Tropheryma whipplei PCR) may be applied to resected valves to aid in diagnosis. Herein, we summarize recent knowledge in this area and propose a microbiologic and pathological algorithm for endocarditis diagnosis. PMID:28659319

Cost analysis in a clinical microbiology laboratory.

PubMed

Brezmes, M F; Ochoa, C; Eiros, J M

2002-08-01

The use of models for business management and cost control in public hospitals has led to a need for microbiology laboratories to know the real cost of the different products they offer. For this reason, a catalogue of microbiological products was prepared, and the costs (direct and indirect) for each product were analysed, along with estimated profitability. All tests performed in the microbiology laboratory of the "Virgen de la Concha" Hospital in Zamora over a 2-year period (73192 tests) were studied. The microbiological product catalogue was designed using homogeneity criteria with respect to procedures used, workloads and costs. For each product, the direct personnel costs (estimated from workloads following the method of the College of American Pathologists, 1992 version), the indirect personnel costs, the direct and indirect material costs and the portion of costs corresponding to the remaining laboratory costs (capital and structural costs) were calculated. The average product cost was 16.05 euros. The average cost of a urine culture (considered, for purposes of this study, as a relative value unit) reached 13.59 euros, with a significant difference observed between positive and negative cultures (negative urine culture, 10.72 euros; positive culture, 29.65 euros). Significant heterogeneity exists, both in the costs of different products and especially in the cost per positive test. The application of a detailed methodology of cost analysis facilitates the calculation of the real cost of microbiological products. This information provides a basic tool for establishing clinical management strategies.

Current and past strategies for bacterial culture in clinical microbiology.

PubMed

Lagier, Jean-Christophe; Edouard, Sophie; Pagnier, Isabelle; Mediannikov, Oleg; Drancourt, Michel; Raoult, Didier

2015-01-01

A pure bacterial culture remains essential for the study of its virulence, its antibiotic susceptibility, and its genome sequence in order to facilitate the understanding and treatment of caused diseases. The first culture conditions empirically varied incubation time, nutrients, atmosphere, and temperature; culture was then gradually abandoned in favor of molecular methods. The rebirth of culture in clinical microbiology was prompted by microbiologists specializing in intracellular bacteria. The shell vial procedure allowed the culture of new species of Rickettsia. The design of axenic media for growing fastidious bacteria such as Tropheryma whipplei and Coxiella burnetii and the ability of amoebal coculture to discover new bacteria constituted major advances. Strong efforts associating optimized culture media, detection methods, and a microaerophilic atmosphere allowed a dramatic decrease of the time of Mycobacterium tuberculosis culture. The use of a new versatile medium allowed an extension of the repertoire of archaea. Finally, to optimize the culture of anaerobes in routine bacteriology laboratories, the addition of antioxidants in culture media under an aerobic atmosphere allowed the growth of strictly anaerobic species. Nevertheless, among usual bacterial pathogens, the development of axenic media for the culture of Treponema pallidum or Mycobacterium leprae remains an important challenge that the patience and innovations of cultivators will enable them to overcome. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Sensor set-up for wireless measurement of automotive rim and wheel parameters in laboratory conditions

NASA Astrophysics Data System (ADS)

Borecki, M.; Prus, P.; Korwin-Pawlowski, M. L.; Rychlik, A.; Kozubel, W.

2017-08-01

Modern rims and wheels are tested at the design and production stages. Tests can be performed in laboratory conditions and on the ride. In the laboratory, complex and costly equipment is used, as for example wheel balancers and impact testers. Modern wheel balancers are equipped with electronic and electro-mechanical units that enable touch-less measurement of dimensions, including precision measurement of radial and lateral wheel run-out, automatic positioning and application of the counterweights, and vehicle wheel set monitoring - tread wear, drift angles and run-out unbalance. Those tests are performed by on-wheel axis measurements with laser distance meters. The impact tester enables dropping of weights from a defined height onto a wheel. Test criteria are the loss of pressure of the tire and generation of cracks in the wheel without direct impact of the falling weights. In the present paper, a set up composed of three accelerometers, a temperature sensor and a pressure sensor is examined as the base of a wheel tester. The sensor set-up configuration, on-line diagnostic and signal transmission are discussed.

Anaerobic Biodegradation Of Methyl tert-Butyl Ether Under Iron-Reducing Conditions In Batch And Continuous-Flow Cultures

EPA Science Inventory

The feasibility of biodegradation of the fuel oxygenate methyl tert-butyl ether (MTBE) under iron-reducing conditions was explored in batch and continuous-flow systems. A porous pot completely-mixed reactor was seeded with diverse cultures and operated under iron-reducing...

The use of propidium iodide to assess excitotoxic neuronal death in primary mixed cortical cultures.

PubMed

Lau, Anthony C; Cui, Hong; Tymianski, Michael

2007-01-01

Neurodegenerative disorders are subjects of intense scrutiny in biomedical research because of their often-debilitating effects. Currently, many laboratories are engaged in developing or testing drugs to prevent neuronal loss in a variety of these pathologies. A key to testing such drugs is the use of a fast, reliable, and easily reproducible model of neurodegeneration and neuroprotection. Our laboratory has previously used propidium iodide (PI) to assess the degree of neurodegeneration and neuroprotection under a variety of conditions. Ultimately, efforts are underway in the laboratory to prevent delayed neuronal loss following acute ischemic insults using drug therapies. It is now believed that a key mechanism of neurodegeneration following acute ischemia or anoxia is a result of excitotoxicity via N-methyl-D-aspartate receptors (NMDARs) and subsequent overproduction of nitric oxide via neuronal nitric oxide synthase (nNOS). Thus, for the purposes of this chapter, the insult used to induce cell death will be various concentrations of NMDA and the compound used to demonstrate neuroprotection will be the nonspecific NOS inhibitor No-nitro-L-arginine methyl ester (L-NAME). Assessment of neuronal death is accomplished by measuring changes in PI fluorescence using a fluorescent plate reader. This chapter will outline the necessary steps required to (1) produce primary mixed cortical cultures, (2) apply PT and NMDA to these cultures, (3) quantify the results obtained from these cultures, and (4) image these cultures in conjunction with Hoechst 33342 and immunocytochemistry using fluorescence microscopy.

Laboratory investigation of spray generation mechanism in wind-wave interaction under strong wind conditions

NASA Astrophysics Data System (ADS)

Kandaurov, Alexander; Troitskaya, Yuliya; Sergeev, Daniil; Ermakova, Olga; Kazakov, Vassily

2015-04-01

The sea spray is considered as a possible mechanism of the reduction of sea surface aerodynamic drag coefficient at hurricane conditions [1]. In this paper the mechanism of generation of spray in the near-surface layer of the atmosphere in a strong wind through the mechanism of «bag-breakup instability» was investigated in laboratory conditions with the help of high-speed video shooting. The laboratory experiments were performed on the Thermostratified Wind-Wave Channel of the IAP RAS (length 10 m, cross section of air channel 0.4 x 0.4 m, wind velocity up to 24 m/s) [2]. Experiments were carried out for the wind speeds from 14 to 22 m/s. In this range spray generation characteristics change dramatically from almost no spray generation to so called catastrophic regime with multiple cascade breakups on each crest. Shooting was performed with High-speed digital camera NAC Memrecam HX-3 in two different setups to obtain both statistical data and detailed spray generation mechanism overview. In first setup bright LED spotlight with mate screen the side of a channel was used for horizontal shadow-method shooting. Camera was placed in semi-submerged box on the opposite side of the channel. Shooting was performed at the distance of 7.5 m from the beginning of the working section. Series of short records of the surface evolution were made at 10 000 fps with 55 to 119 µm/px scale revealed the dominant mechanism of spray generation - bag-breakup instability. Sequences of high resolution images allowed investigating the details of this "bags" evolution. Shadow method provided better image quality for such conditions than side illumination and fluorescence methods. To obtain statistical data on "bags" sizes and densities vertical shadow method was used. Submerged light box was created with two 300 W underwater lamps and mate screen places at the fetch of 6.5 m. Long records (up to 8 seconds) were made with 4500 fps at 124-256 µm/px scales. Specially developed software

Use of CHROMagar Candida for genital specimens in the diagnostic laboratory.

PubMed Central

Houang, E T; Chu, K C; Koehler, A P; Cheng, A F

1997-01-01

OBJECTIVE: To evaluate CHROMagar Candida (CA), a new yeast differential medium, for yeast isolation in a clinical laboratory for the routine examination of high vaginal swabs. METHODS: Results of high vaginal swab cultures processed in a standard manner on plates containing equal halves of Sabouraud dextrose agar (SDA) and CA were compared. Non-Candida albicans yeast isolates were further speciated with API 20C AUX or API 32C. To assess the ease of use of CA, laboratory staff lacking in experience of the medium were asked to identify 23 unlabelled yeast cultures on CA by referring to six labelled reference plates. RESULTS: Of the 1784 swab cultures processed, yeasts were isolated from 373 SDA and 368 CA. Of the 78 non-albicans isolates further speciated, CA identified correctly all cultures of C krusei and C tropicalis, and 82% of C glabrata. All the 38 inexperienced laboratory staff achieved 100% accuracy for C albicans and over 90% for C krusei and C tropicalis. CONCLUSIONS: CA is a satisfactory isolation medium for genital specimens, allowing immediate and correct identification of the commonly encountered yeasts and easy recognition of mixed cultures. Images PMID:9306935

Response of Xylella fastidiosa to zinc: decreased culturability, increased exopolysaccharide production, and formation of resilient biofilms under flow conditions.

PubMed

Navarrete, Fernando; De La Fuente, Leonardo

2014-02-01

The bacterial plant pathogen Xylella fastidiosa produces biofilm that accumulates in the host xylem vessels, affecting disease development in various crops and bacterial acquisition by insect vectors. Biofilms are sensitive to the chemical composition of the environment, and mineral elements being transported in the xylem are of special interest for this pathosystem. Here, X. fastidiosa liquid cultures were supplemented with zinc and compared with nonamended cultures to determine the effects of Zn on growth, biofilm, and exopolysaccharide (EPS) production under batch and flow culture conditions. The results show that Zn reduces growth and biofilm production under both conditions. However, in microfluidic chambers under liquid flow and with constant bacterial supplementation (closer to conditions inside the host), a dramatic increase in biofilm aggregates was seen in the Zn-amended medium. Biofilms formed under these conditions were strongly attached to surfaces and were not removed by medium flow. This phenomenon was correlated with increased EPS production in stationary-phase cells grown under high Zn concentrations. Zn did not cause greater adhesion to surfaces by individual cells. Additionally, viability analyses suggest that X. fastidiosa may be able to enter the viable but nonculturable state in vitro, and Zn can hasten the onset of this state. Together, these findings suggest that Zn can act as a stress factor with pleiotropic effects on X. fastidiosa and indicate that, although Zn could be used as a bactericide treatment, it could trigger the undesired effect of stronger biofilm formation upon reinoculation events.

Response of Xylella fastidiosa to Zinc: Decreased Culturability, Increased Exopolysaccharide Production, and Formation of Resilient Biofilms under Flow Conditions

PubMed Central

Navarrete, Fernando

2014-01-01

The bacterial plant pathogen Xylella fastidiosa produces biofilm that accumulates in the host xylem vessels, affecting disease development in various crops and bacterial acquisition by insect vectors. Biofilms are sensitive to the chemical composition of the environment, and mineral elements being transported in the xylem are of special interest for this pathosystem. Here, X. fastidiosa liquid cultures were supplemented with zinc and compared with nonamended cultures to determine the effects of Zn on growth, biofilm, and exopolysaccharide (EPS) production under batch and flow culture conditions. The results show that Zn reduces growth and biofilm production under both conditions. However, in microfluidic chambers under liquid flow and with constant bacterial supplementation (closer to conditions inside the host), a dramatic increase in biofilm aggregates was seen in the Zn-amended medium. Biofilms formed under these conditions were strongly attached to surfaces and were not removed by medium flow. This phenomenon was correlated with increased EPS production in stationary-phase cells grown under high Zn concentrations. Zn did not cause greater adhesion to surfaces by individual cells. Additionally, viability analyses suggest that X. fastidiosa may be able to enter the viable but nonculturable state in vitro, and Zn can hasten the onset of this state. Together, these findings suggest that Zn can act as a stress factor with pleiotropic effects on X. fastidiosa and indicate that, although Zn could be used as a bactericide treatment, it could trigger the undesired effect of stronger biofilm formation upon reinoculation events. PMID:24271184

Laboratory evaluation and application of microwave absorption properties under simulated conditions for planetary atmospheres

NASA Technical Reports Server (NTRS)

Steffes, P. G.

1985-01-01

Radio absorptivity data for planetary atmospheres obtained from spacecraft radio occultation experiments and Earth-based radio astronomical observations can be used to infer abundances of microwave absorbing atmospheric constituents in those atmospheres, as long as reliable information regarding the microwave absorbing properties of potential constituents is available. The use of theoretically-derived microwave absorption properties for such atmospheric constituents, or laboratory measurements of such properties under environmental conditions which are significantly different than those of the planetary atmosphere being studied, often lead to significant misinterpretation of available opacity data. Steffes and Eshleman showed that under environmental conditions corresponding to the middle atmosphere of Venus, the microwave absorption due to atmospheric SO2 was 50 percent greater than that calculated from Van Vleck-Weiskopff theory. Similarly, the opacity from gaseous H2SO4 was found to be a factor of 7 greater than theoretically predicted for conditions of the Venus middle atmosphere. The recognition of the need to make such measurements over a range of temperatures and pressures which correspond to the periapsis altitudes of radio occultation experiments, and over a range of frequencies which correspond to both radio occultation experiments and radio astronomical observations, has led to the development of a facility at Georgia Tech which is capable of making such measurements.

Development of an algorithm for production of inactivated arbovirus antigens in cell culture

PubMed Central

Goodman, C.H.; Russell, B.J.; Velez, J.O.; Laven, J.J.; Nicholson, W.L; Bagarozzi, D.A.; Moon, J.L.; Bedi, K.; Johnson, B.W.

2015-01-01

Arboviruses are medically important pathogens that cause human disease ranging from a mild fever to encephalitis. Laboratory diagnosis is essential to differentiate arbovirus infections from other pathogens with similar clinical manifestations. The Arboviral Diseases Branch (ADB) reference laboratory at the CDC Division of Vector-Borne Diseases (DVBD) produces reference antigens used in serological assays such as the virus-specific immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA). Antigen production in cell culture has largely replaced the use of suckling mice; however, the methods are not directly transferable. The development of a cell culture antigen production algorithm for nine arboviruses from the three main arbovirus families, Flaviviridae, Togaviridae, and Bunyaviridae, is described here. Virus cell culture growth and harvest conditions were optimized, inactivation methods were evaluated, and concentration procedures were compared for each virus. Antigen performance was evaluated by the MAC-ELISA at each step of the procedure. The antigen production algorithm is a framework for standardization of methodology and quality control; however, a single antigen production protocol was not applicable to all arboviruses and needed to be optimized for each virus. PMID:25102428

Effects of medium components and culture conditions on mycelial biomass and the production of bioactive ingredients in submerged culture of Xylaria nigripes (Ascomycetes), a Chinese medicinal fungus.

PubMed

Chen, Jian-Zhi; Lo, Hui-Chen; Lin, Fang-Yi; Chang, Shih-Liang; Hsieh, Changwei; Liang, Zeng-Chin; Ho, Wai-Jane; Hsu, Tai-Hao

2014-01-01

The optimal culture conditions were investigated to maximize the production of mycelial biomass and bioactive ingredients in submerged cultivation of Xylaria nigripes, a Chinese medicinal fungus. The one-factor-at-a-time method was used to explore the effects of medium components, including carbon, nitrogen, mineral sources, and initial pH of the medium and environmental factors, such as culture temperature and rotation speed, on mycelial growth and production of bioactive ingredients. The results indicated that the optimal culture temperature and rotation speed were 25°C and 100 rpm in a medium with 20 g fructose, 6 g yeast extract, and 2 g magnesiun sulfate heptahydrate as carbon, nitrogen, and mineral sources, respectively, in 1 L distilled water with an initial medium pH of 5.5. With optimal medium components and conditions of cultivation, the maximal production of mycelial biomass was 6.64 ± 0.88 g/L, with maximal production of bioactive ingredients such as extracellular polysaccharides (2.36 ± 0.18 mg/mL), intracellular polysaccharides (2.38 ± 0.07 mg/g), adenosine (43.27 ± 2.37 mg/g), total polyphenols (36.57 ± 1.36 mg/g), and triterpenoids (31.29 ± 1.17 mg/g) in a shake flask culture. These results suggest that different bioactive ingredients including intracellular polysaccharides, adenosine, total polyphenols and triterpenoids in mycelia and extracellular polysaccharides in broth can be obtained from one simple medium for submerged cultivation of X. nigripes.

Load dissipation by corn residue on tilled soil in laboratory and field-wheeling conditions.

PubMed

Reichert, José M; Brandt, André A; Rodrigues, Miriam F; Reinert, Dalvan J; Braida, João A

2016-06-01

Crop residues may partially dissipate applied loads and reduce soil compaction. We evaluated the effect of corn residue on energy-applied dissipation during wheeling. The experiment consisted of a preliminary laboratory test and a confirmatory field test on a Paleaudalf soil. In the laboratory, an adapted Proctor test was performed with three energy levels, with and without corn residue. Field treatments consisted of three 5.1 Mg tractor wheeling intensities (0, 2, and 6), with and without 12 Mg ha(-1) corn residue on the soil surface. Corn residue on the soil surface reduced soil bulk density in the adapted Proctor test. By applying energy of 52.6 kN m m(-3) , soil dissipated 2.98% of applied energy, whereas with 175.4 kN m m(-3) a dissipation of 8.60% was obtained. This result confirms the hypothesis that surface mulch absorbs part of the compaction effort. Residue effects on soil compaction observed in the adapted Proctor test was not replicated under subsoiled soil field conditions, because of differences in applied pressure and soil conditions (structure, moisture and volume confinement). Nevertheless, this negative result does not mean that straw has no effect in the field. Such effects should be measured via stress transmission and compared to soil load-bearing capacity, rather than on bulk deformations. Wheeling by heavy tractor on subsoiled soil increased compaction, independently of surface residue. Two wheelings produced a significantly increase, but six wheelings did not further increase compaction. Reduced traffic intensity on recently tilled soil is necessary to minimize soil compaction, since traffic intensity show a greater effect than surface mulch on soil protection from excessive compaction. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

DO TIE LABORATORY BASED METHODS REALLY REFLECT FIELD CONDITIONS

EPA Science Inventory

Sediment Toxicity Identification and Evaluation (TIE) methods have been developed for both interstitial waters and whole sediments. These relatively simple laboratory methods are designed to identify specific toxicants or classes of toxicants in sediments; however, the question ...

Activity, aggression, and habitat use of ruffe (Gymnocephalus cernuus) and round goby (Apollonia melanostoma) under laboratory conditions

USGS Publications Warehouse

Savino, J.F.; Riley, S.C.; Holuszko, M.J.

2007-01-01

Potential negative ecological interactions between ruffe Gymnocephalus cernuus and round gobyApollonia melanostoma (formerly Neogobius melanostomus) might affect the colonization dynamics of these invasive species where they are sympatric in the Great Lakes. In order to determine the potential for ecological interactions between these species, we examined the activity, aggression, and habitat use of round gobies and ruffe in single species and mixed species laboratory experiments. Trials included conditions in which food was concentrated (in light or darkness) or scattered. Results showed that ruffe were more active than gobies, particularly when food was scattered. Activity of both species was significantly lower during darkness. Round gobies were significantly more aggressive than ruffe, and total aggression was lower in mixed species trials. Habitat use by ruffe and round gobies overlapped considerably, but we observed significant differences between species in their use of specific habitats that depended on experimental conditions. Overall, ruffe used open habitats more often than did round gobies, primarily when food was scattered. Round gobies used rocks significantly more frequently than did ruffe, but their use of rock habitat decreased during dark conditions. Ruffe were found more often in plant habitats and less often near the wall of the pool in trials during daylight with concentrated food. Activity and habitat use of ruffe and round goby did not significantly differ between single and mixed species trials. Overall, we found little evidence for negative ecological interactions between ruffe and round goby in these laboratory experiments.

Keratinocytes propagated in serum-free, feeder-free culture conditions fail to form stratified epidermis in a reconstituted skin model.

PubMed

Lamb, Rebecca; Ambler, Carrie A

2013-01-01

Primary human epidermal stem cells isolated from skin tissues and subsequently expanded in tissue culture are used for human therapeutic use to reconstitute skin on patients and to generate artificial skin in culture for academic and commercial research. Classically, epidermal cells, known as keratinocytes, required fibroblast feeder support and serum-containing media for serial propagation. In alignment with global efforts to remove potential animal contaminants, many serum-free, feeder-free culture methods have been developed that support derivation and growth of these cells in 2-dimensional culture. Here we show that keratinocytes grown continually in serum-free and feeder-free conditions were unable to form into a stratified, mature epidermis in a skin equivalent model. This is not due to loss of cell potential as keratinocytes propagated in serum-free, feeder-free conditions retain their ability to form stratified epidermis when re-introduced to classic serum-containing media. Extracellular calcium supplementation failed to improve epidermis development. In contrast, the addition of serum to commercial, growth media developed for serum-free expansion of keratinocytes facilitated 3-dimensional stratification in our skin equivalent model. Moreover, the addition of heat-inactivated serum improved the epidermis structure and thickness, suggesting that serum contains factors that both aid and inhibit stratification.

Keratinocytes Propagated in Serum-Free, Feeder-Free Culture Conditions Fail to Form Stratified Epidermis in a Reconstituted Skin Model