The content of indigo precursors in Isatis tinctoria leaves--a comparative study of selected accessions and post-harvest treatments.

PubMed

Oberthür, Christine; Graf, Heidemarie; Hamburger, Matthias

2004-12-01

We recently clarified the nature of indigo precursors in woad (Isatis tinctoria L.), by identifying the major indoxyl glycoside as isatan A (indoxyl-3-O-(6'-O-malonyl-beta-D-ribohexo-3-ulopyranoside)), and by correcting the structure of the related isatan B (indoxyl-3-O-beta-D-ribohexo-3-ulopyranoside). A quantitative densitometric assay for isatans A and B, and indican, was established and validated. HPTLC separation on silica gel was followed by densitometric analysis of indigoid pigments formed after treatment with dilute acid or base. The seasonal variation of indoxyl glycosides in woad leaves was investigated with first-year plants (rosette stage) of five defined I. tinctoria L. and one I. indigotica L. accessions. Isatan A content reached up to 7.6% of dry weight in I. tinctoria, and up to 21.8% in I. indigotica. The influence of various post-harvest treatments was studied. High concentrations of isatans A and B were found in freeze-dried leaf samples, whereas the content of indican was lowest. Conventional drying at ambient or 40 degrees C led to complete disappearance of isatans A and B. The concentration of indican, in contrast, was 3- to 5-fold higher in leaf samples submitted to drying at ambient and 40 degrees C, respectively.

Estimation of capsaicin through scanning densitometry and evaluation of different varieties of capsicum in India.

PubMed

Gantait, Arunava; Maji, Amal; Barman, Tapu; Banerji, Pratim; Venkatesh, P; Mukherjee, Pulok K

2012-01-01

Capsicum annuum L. (family: Solanaceae) possesses therapeutic benefits for the treatment of rheumatism, neuropathy, psoriasis, flatulence and so on. In this study fruits of four different varieties of C. annuum from four different geographical regions in India were evaluated based on their total content of capsaicin. Ethanol extracts of the fruits were used. HPTLC plates were developed in a mobile phase containing benzene, ethyl acetate and methanol (75:20:5). Densitometric scanning was performed at a wavelength of 283 nm in the absorbance mode. The calibration curve was described by the equation Y=393.587+3.836*X with a correlation coefficient (r) of 0.99890. The content of capsaicin in Nagaland, Manipur, West Bengal and Shimla varieties was found to be 3.71%, 1.78%, 0.54% and 0.06%, respectively. The developed densitometric method was found to be specific, accurate and precise. A recovery study and precision showed low levels of %RSD values. The linearity range of the curve for capsaicin was found to be 300-900 ng per spot. The limit of detection and the limit of quantification values were determined to be 31 and 94 ng, respectively, proving the sensitivity of the method. Thus the method can be used to control the total content of capsaicin on an industrial scale.

Validation Thin Layer Chromatography for the Determination of Acetaminophen in Tablets and Comparison with a Pharmacopeial Method

PubMed Central

Pyka, Alina; Budzisz, Marika; Dołowy, Małgorzata

2013-01-01

Adsorption thin layer chromatography (NP-TLC) with densitometry has been established for the identification and the quantification of acetaminophen in three leading commercial products of pharmaceutical tablets coded as brand: P1 (Product no. 1), P2 (Product no. 2), and P3 (Product no. 3). Applied chromatographic conditions have separated acetaminophen from its related substances, namely, 4-aminophenol and and 4′-chloroacetanilide. UV densitometry was performed in absorbance mode at 248 nm. The presented method was validated by specificity, range, linearity, accuracy, precision, detection limit, quantitative limit, and robustness. The TLC-densitometric method was also compared with a pharmacopeial UV-spectrophotometric method for the assay of acetaminophen, and the results confirmed statistically that the NP-TLC-densitometric method can be used as a substitute method. It could be said that the validated NP-TLC-densitometric method is suitable for the routine analysis of acetaminophen in quantity control laboratories. PMID:24063006

Validation of a reversed phase high performance thin layer chromatographic-densitometric method for secoisolariciresinol diglucoside determination in flaxseed.

PubMed

Coran, Silvia A; Bartolucci, Gianluca; Bambagiotti-Alberti, Massimo

2008-10-17

The validation of a HPTLC-densitometric method for the determination of secoisolariciresinol diglucoside (SDG) in flaxseed was performed improving the reproducibility of a previously reported HPTLC densitometric procedure by the use of fully wettable reversed phase plates (silica gel 60 RP18W F(254S), 10cmx10cm) with MeOH:HCOOH 0.1% (40:60, v/v) mobile phase. The analysis required only the alkaline hydrolysis in aqueous medium of undefatted samples and densitometry at 282nm of HPTLC runs. The method was validated following the protocol proposed by the Société Francaise des Sciences et Techniques Pharmaceutiques (SFSTP) giving rise to a dependable and high throughput procedure well suited to routine application. SDG was quantified in the range of 321-1071ng with RSD of repeatability and intermediate precision not exceeding 3.61% and accuracy inside the acceptance limits. Flaxseed of five cultivars of different origin was elected as test-bed.

Quantitative analysis of γ-oryzanol content in cold pressed rice bran oil by TLC-image analysis method.

PubMed

Sakunpak, Apirak; Suksaeree, Jirapornchai; Monton, Chaowalit; Pathompak, Pathamaporn; Kraisintu, Krisana

2014-02-01

To develop and validate an image analysis method for quantitative analysis of γ-oryzanol in cold pressed rice bran oil. TLC-densitometric and TLC-image analysis methods were developed, validated, and used for quantitative analysis of γ-oryzanol in cold pressed rice bran oil. The results obtained by these two different quantification methods were compared by paired t-test. Both assays provided good linearity, accuracy, reproducibility and selectivity for determination of γ-oryzanol. The TLC-densitometric and TLC-image analysis methods provided a similar reproducibility, accuracy and selectivity for the quantitative determination of γ-oryzanol in cold pressed rice bran oil. A statistical comparison of the quantitative determinations of γ-oryzanol in samples did not show any statistically significant difference between TLC-densitometric and TLC-image analysis methods. As both methods were found to be equal, they therefore can be used for the determination of γ-oryzanol in cold pressed rice bran oil.

Quantitative analysis of γ-oryzanol content in cold pressed rice bran oil by TLC-image analysis method

PubMed Central

Sakunpak, Apirak; Suksaeree, Jirapornchai; Monton, Chaowalit; Pathompak, Pathamaporn; Kraisintu, Krisana

2014-01-01

Objective To develop and validate an image analysis method for quantitative analysis of γ-oryzanol in cold pressed rice bran oil. Methods TLC-densitometric and TLC-image analysis methods were developed, validated, and used for quantitative analysis of γ-oryzanol in cold pressed rice bran oil. The results obtained by these two different quantification methods were compared by paired t-test. Results Both assays provided good linearity, accuracy, reproducibility and selectivity for determination of γ-oryzanol. Conclusions The TLC-densitometric and TLC-image analysis methods provided a similar reproducibility, accuracy and selectivity for the quantitative determination of γ-oryzanol in cold pressed rice bran oil. A statistical comparison of the quantitative determinations of γ-oryzanol in samples did not show any statistically significant difference between TLC-densitometric and TLC-image analysis methods. As both methods were found to be equal, they therefore can be used for the determination of γ-oryzanol in cold pressed rice bran oil. PMID:25182282

Nuclear morphometry in flat epithelial atypia of the breast as a predictor of malignancy: a digital image-based histopathologic analysis.

PubMed

Williams, Phillip A; Djordjevic, Bojana; Ayroud, Yasmine; Islam, Shahidul; Gravel, Denis; Robertson, Susan J; Parra-Herran, Carlos

2014-12-01

To identify morphometric features unique to flat epithelial atypia associated with cancer using digital image analysis. Cases with diagnosis of flat epithelial atypia were retrieved and divided into 2 groups: flat epithelial atypia associated with invasive or in situ carcinoma (n = 31) and those without malignancy (n = 27). Slides were digitally scanned. Nuclear features were analyzed on representative images at 20x magnification using digital morphometric software. Parameters related to nuclear shape and size (diameter, perimeter) were similar in both groups. However, cases with malignancy had significantly higher densitometric green (p = 0.02), red (p = 0.03), and grey (p = 0.02) scale levels as compared to cases without cancer. A mean grey densitometric level > 119.45 had 71% sensitivity and 70.4% specificity in detecting cases with concomitant carcinoma. Morphometry of features related to nuclear staining appears to be useful in predicting risk of concurrent malignancy in patients with flat epithelial atypia, when added to a comprehensive histopathologic evaluation.

Optimization of analytical parameters for inferring relationships among Escherichia coli isolates from repetitive-element PCR by maximizing correspondence with multilocus sequence typing data.

PubMed

Goldberg, Tony L; Gillespie, Thomas R; Singer, Randall S

2006-09-01

Repetitive-element PCR (rep-PCR) is a method for genotyping bacteria based on the selective amplification of repetitive genetic elements dispersed throughout bacterial chromosomes. The method has great potential for large-scale epidemiological studies because of its speed and simplicity; however, objective guidelines for inferring relationships among bacterial isolates from rep-PCR data are lacking. We used multilocus sequence typing (MLST) as a "gold standard" to optimize the analytical parameters for inferring relationships among Escherichia coli isolates from rep-PCR data. We chose 12 isolates from a large database to represent a wide range of pairwise genetic distances, based on the initial evaluation of their rep-PCR fingerprints. We conducted MLST with these same isolates and systematically varied the analytical parameters to maximize the correspondence between the relationships inferred from rep-PCR and those inferred from MLST. Methods that compared the shapes of densitometric profiles ("curve-based" methods) yielded consistently higher correspondence values between data types than did methods that calculated indices of similarity based on shared and different bands (maximum correspondences of 84.5% and 80.3%, respectively). Curve-based methods were also markedly more robust in accommodating variations in user-specified analytical parameter values than were "band-sharing coefficient" methods, and they enhanced the reproducibility of rep-PCR. Phylogenetic analyses of rep-PCR data yielded trees with high topological correspondence to trees based on MLST and high statistical support for major clades. These results indicate that rep-PCR yields accurate information for inferring relationships among E. coli isolates and that accuracy can be enhanced with the use of analytical methods that consider the shapes of densitometric profiles.

Simultaneous Determination of Crypto-Chlorogenic Acid, Isoquercetin, and Astragalin Contents in Moringa oleifera Leaf Extracts by TLC-Densitometric Method

PubMed Central

Vongsak, Boonyadist; Sithisarn, Pongtip; Gritsanapan, Wandee

2013-01-01

Moringa oleifera Lamarck (Moringaceae) is used as a multipurpose medicinal plant for the treatment of various diseases. Isoquercetin, astragalin, and crypto-chlorogenic acid have been previously found to be major active components in the leaves of this plant. In this study, a thin-layer-chromatography (TLC-)densitometric method was developed and validated for simultaneous quantification of these major components in the 70% ethanolic extracts of M. oleifera leaves collected from 12 locations. The average amounts of crypto-chlorogenic acid, isoquercetin, and astragalin were found to be 0.0473, 0.0427, and 0.0534% dry weight, respectively. The method was validated for linearity, precision, accuracy, limit of detection, limit of quantitation, and robustness. The linearity was obtained in the range of 100–500 ng/spot with a correlation coefficient (r) over 0.9961. Intraday and interday precisions demonstrated relative standard deviations of less than 5%. The accuracy of the method was confirmed by determining the recovery. The average recoveries of each component from the extracts were in the range of 98.28 to 99.65%. Additionally, the leaves from Chiang Mai province contained the highest amounts of all active components. The proposed TLC-densitometric method was simple, accurate, precise, and cost-effective for routine quality controlling of M. oleifera leaf extracts. PMID:23533530

Prevalence of vertebral fracture and densitometric osteoporosis in Spanish adult men: The Camargo Cohort Study.

PubMed

Olmos, José M; Hernández, José L; Martínez, Josefina; Pariente, Emilio; Castillo, Jesús; Prieto-Alhambra, Daniel; González-Macías, Jesús

2018-01-01

The aim of this study was to assess the prevalence of densitometric osteoporosis and vertebral fractures in Spanish men aged ≥50 years, and to study how the relationship between them may change depending on how osteoporosis is diagnosed. A community-based population of 1003 men aged ≥50 years was studied. Bone mineral density (BMD) was measured by DXA at the lumbar spine, femoral neck and total hip. Vertebral fractures were assessed by lateral thoracic and lumbar spine radiographs. The prevalence of osteoporosis was estimated with both the World Health Organization (WHO) (T-score of <-2.5 at the femoral neck, calculated using the young white female normal reference database) and the National Osteoporosis Foundation (NOF) criteria (T-score of <-2.5 at the femoral neck, total hip or lumbar spine, calculated using the young white male normal reference database). The prevalence of osteoporosis using the WHO criterion was 1.1% and using the NOF criterion was 13%, while that of vertebral fractures was 21.3%. The area under the curve (AUC) for the relationship between BMD and vertebral fracture prevalence was 0.64. The odds ratio for osteoporosis using the WHO definition was 2.57 (p = 0.13), and 1.78 (p = 0.007) using the NOF definition. Vertebral fracture prevalence rose with age. The prevalence of osteoporosis increased only moderately in men aged >70 years with the WHO criterion, and showed no change using the NOF definition. The prevalence of osteoporosis in Spanish men using the WHO definition is too small to have any meaningful clinical use. Although the figure is higher using the NOF definition, it would seem that population-based studies of BMD in men are of questionable value.

Spectrofluorometry, thin layer chromatography, and column high-performance liquid chromatography determination of rabeprazole sodium in the presence of its acidic and oxidized degradation products.

PubMed

Osman, Afaf Osman; Osman, Afaf; Osman, Mohamed

2009-01-01

The objective of this study is to develop validated stability-indicating spectrofluorometric, TLC-densitometric, and HPLC methods for the determination of rabeprazole sodium and its degradation products. The first method was based on measuring the fluorescence intensity of the drug at 416 and 311 nm for the emission and at 320 and 274 nm for the excitation for acid and oxidized solutions, respectively. The second method was based on the separation of the drug from its acidic and oxidized degradation products followed by densitometric measurement of the intact drug spot at 284 nm. The separation was carried out on Fluka TLC sheets of silica gel 60 F254 using isopropyl alcohol--30% ammonia (80 + 2, v/v) mobile phase. The third method was based on HPLC separation of rabeprazole sodium from its acidic and oxidized degradation products on a reversed-phase Waters Nova-Pak C18 column using 0.05 M potassium dihydrogen phosphate-methanol-acetonitrile (5 + 3 + 2, v/v/v) pH 7 +/- 0.2 mobile phase. The proposed procedures were successfully applied for the determination of rabeprazole sodium in pure form, laboratory-prepared mixtures, tablet, and expired batch. The obtained results were statistically compared with those of a reported method and validated according to United States Pharmacopeia guidelines. Two main acidic degradation products of the drug were separated and subjected to IR spectrometry and MS to confirm their structures, and the schemes for their formation were elucidated.

Quantitative data standardization of X-ray based densitometry methods

NASA Astrophysics Data System (ADS)

Sergunova, K. A.; Petraikin, A. V.; Petrjajkin, F. A.; Akhmad, K. S.; Semenov, D. S.; Potrakhov, N. N.

2018-02-01

In the present work is proposed the design of special liquid phantom for assessing the accuracy of quantitative densitometric data. Also are represented the dependencies between the measured bone mineral density values and the given values for different X-ray based densitometry techniques. Shown linear graphs make it possible to introduce correction factors to increase the accuracy of BMD measurement by QCT, DXA and DECT methods, and to use them for standardization and comparison of measurements.

A densitometric analysis of IIaO film flown aboard the space shuttle transportation system STS #3, 7, and 8

NASA Technical Reports Server (NTRS)

Hammond, Ernest C., Jr.

1989-01-01

Since the United States of America is moving into an age of reusable space vehicles, both electronic and photographic materials will continue to be an integral part of the recording techniques available. Film as a scientifically viable recording technique in astronomy is well documented. There is a real need to expose various types of films to the Shuttle environment. Thus, the main objective was to look at the subtle densitometric changes of canisters of IIaO film that was placed aboard the Space Shuttle 3 (STS-3).

A densitometric analysis of commercial 35mm films

NASA Technical Reports Server (NTRS)

Hammond, Ernest C., Jr.; Ruffin, Christopher, III

1989-01-01

IIaO films have been subjected to various sensitometric tests. The have included thermal and aging effects and reciprocity failure studies. In order to compare the special IIaO film with popular brands of 35 mm films and their possible use in astrophotography, Agfa, Fuji and Kodak print and slide formats, as well as black and white and color formats, were subjected to sensitometric, as well as densitometric analysis. A scanning electron microscope was used to analyze grain structure size, and shape as a function of both speed and brand. Preliminary analysis of the grain structure using an ISI-SS40 scanning electron microscope indicates that the grain sizes for darker densities are much larger than the grain size for lighter densities. Researchers analyze the scanning electron microscope findings of the various grains versus densities as well as enhancement of the grains, using the IP-8500 Digital Image Processor.

Generalized metabolic bone disease in Neurofibromatosis type I

USDA-ARS?s Scientific Manuscript database

Skeletal abnormalities are a recognized component of Neurofibromatosis type I (NF1), but a generalized metabolic bone defect in NF1 has not been fully characterized thus far. The purpose of this study was to characterize at the densitometric, biochemical, and pathological level the bone involvement ...

The influence of exercise on bone atrophy

NASA Technical Reports Server (NTRS)

1978-01-01

The relationship between the skeletal system, the muscular system, and exercise in bed rest studies are described. The regime of exercises performed, the mineral balance data derived, and the bone densitometric data obtained are discussed. A brief review of some of the histological results are also given.

Scan-rescan reproducibility of CT densitometric measures of emphysema

NASA Astrophysics Data System (ADS)

Chong, D.; van Rikxoort, E. M.; Kim, H. J.; Goldin, J. G.; Brown, M. S.

2011-03-01

This study investigated the reproducibility of HRCT densitometric measures of emphysema in patients scanned twice one week apart. 24 emphysema patients from a multicenter study were scanned at full inspiration (TLC) and expiration (RV), then again a week later for four scans total. Scans for each patient used the same scanner and protocol, except for tube current in three patients. Lung segmentation with gross airway removal was performed on the scans. Volume, weight, mean lung density (MLD), relative area under -950HU (RA-950), and 15th percentile (PD-15) were calculated for TLC, and volume and an airtrapping mask (RA-air) between -950 and -850HU for RV. For each measure, absolute differences were computed for each scan pair, and linear regression was performed against volume difference in a subgroup with volume difference <500mL. Two TLC scan pairs were excluded due to segmentation failure. The mean lung volumes were 5802 +/- 1420mL for TLC, 3878 +/- 1077mL for RV. The mean absolute differences were 169mL for TLC volume, 316mL for RV volume, 14.5g for weight, 5.0HU for MLD, 0.66p.p. for RA-950, 2.4HU for PD-15, and 3.1p.p. for RA-air. The <500mL subgroup had 20 scan pairs for TLC and RV. The R2 values were 0.8 for weight, 0.60 for MLD, 0.29 for RA-950, 0.31 for PD-15, and 0.64 for RA-air. Our results indicate that considerable variability exists in densitometric measures over one week that cannot be attributed to breathhold or physiology. This has implications for clinical trials relying on these measures to assess emphysema treatment efficacy.

Serum Concentration of Macro-, Micro-, and Trace Elements in Silver Fox (Vulpes vulpes) and Their Interrelationships with Morphometric, Densitometric, and Mechanical Properties of the Mandible.

PubMed

Tatara, Marcin R; Łuszczewska-Sierakowska, Iwona; Krupski, Witold

2017-12-20

The optimal content of macro-, micro-, and trace elements in tissues ensures proper systemic growth and development and optimal health status in animals and humans. However, very little is known on the elemental content in the plasma compartment in Silver fox. The aim of this study was to determine the content of selected elements in serum obtained from 8-month-old female (N = 8) and male (N = 7) silver foxes. Moreover, relationships of the evaluated elements with the morphological, densitometric, and mechanical parameters of the mandible were determined. Serum content of 12 different elements was measured using inductively coupled plasma-atomic emission spectrometry. The morphometric and densitometric properties of the mandible were determined using quantitative computed tomography method, while mechanical endurance was tested using a three-point bending test. Serum concentration of calcium was significantly higher by 20% in male foxes (P = 0.01), while manganese concentration was significantly lower in males by over 17% (P = 0.03). Positive correlations of serum concentration of calcium, phosphorus, and magnesium with the morphological traits of the mandible such as weight, length, and bone volume were stated (P < 0.05). In the group of elements playing regulatory functions, the positive relationships between serum concentrations of selenium, chromium, manganese, copper, and cobalt were found (P < 0.05). The elaborated experimental model may serve for further studies on foxes, especially focused on nutritional factors affecting elemental homeostasis, whole-body metabolism, and systemic growth and development. Daily diet formulation and precise delivery for farm foxes, together with relatively large animal population maintained at the same environmental conditions, regularly subjected to slaughter procedure, enable economical experimentation with various dietary and pharmacological manipulations.

Impact of osteoporosis and vertebral fractures on quality-of-life. a population-based study in Valencia, Spain (The FRAVO Study).

PubMed

Sanfélix-Genovés, José; Hurtado, Isabel; Sanfélix-Gimeno, Gabriel; Reig-Molla, Begoña; Peiró, Salvador

2011-04-06

To describe the health related quality of life in a population sample of postmenopausal women over the age of 50 and resident in the city of Valencia (Spain), according to the presence/absence of osteoporosis and the severity of prevalent morphometric vertebral fractures. A cross-sectional age-stratified population-based sample of 804 postmenopausal women of 50 years of age and older were assessed with the SF-12 questionnaire. Information about demographic features, lifestyle, clinical features, educational level, anti-osteoporotic and other treatments, comorbidities and risk factors for osteoporosis were collected using an interviewer-administered questionnaire and densitometric evaluation of spine and hip and spine x-rays were carried out. In the non-adjusted analysis, mild and moderate-severe vertebral fractures were associated with decreased scores in the SF-12 Physical Component Summary (PCS) but not in the Mental Component Summary (MCS), while densitometric osteoporosis with no accompanying fracture was not associated with a worse health related quality of life. In multivariate analysis worse PCS scores were associated to the age groups over 70 (-2.43 for 70-74 group and -2.97 for 75 and older), chronic conditions (-4.66, -6.79 and -11.8 according to the presence of 1, 2 or at least 3 conditions), obesity (-5.35), peripheral fracture antecedents (-3.28), hypoestrogenism antecedents (-2.61) and the presence of vertebral fracture (-2.05). After adjusting for confounding factors, the physical components of health related quality of life were significantly lower in women with prevalent osteoporotic vertebral fractures than in women--osteoporotic or not--without vertebral fractures.

X-RAY DENSITOMETRY OF NORWAY SPRUCE SUBFOSSIL WOOD FROM THE AUSTRIAN ALPS

PubMed Central

KŁUSEK, MARZENA; GRABNER, MICHAEL

2016-01-01

The processing of subfossil wood poses some difficulties in densitometric research. Problems arise because of the physio-chemical changes of wood occurring in the sedimentation environment. Subfossil wood modification can result from the uptake of mineral and organic substances into the wood tissue. It can also occur as the effect of microbiological degradation of wood. The goal of this study was to identify the appropriate method of subfossil wood preparation for the densitometric research. For this purpose the wood of Norway spruce from Lake Schwarzensee was subjected to extraction in deionized water, acetone and diluted acetic acid. The application of acetic acid did not significantly influence the density of the wood and acetone seemed to be too aggressive. The best result was obtained by rinsing the samples in cold de-ionized water. This extraction procedure allowed removal of unwanted water-soluble, organic and inorganic compounds from wood and simultaneously did not lead to the degradation of subfossil samples. PMID:27158247

Interrelationships Between Morphometric, Densitometric, and Mechanical Properties of Teeth in 5-Month-Old Polish Merino Sheep.

PubMed

Tatara, Marcin R; Szabelska, Anna; Krupski, Witold; Tymczyna, Barbara; Łuszczewska-Sierakowska, Iwona; Bieniaś, Jarosław; Ostapiuk, Monika

2018-06-01

Interrelationships between morphological, densitometric, and mechanical properties of deciduous mandibular teeth (incisors, canine, second premolar) were investigated. To perform morphometric, densitometric, and mechanical analyses, teeth were obtained from 5-month-old sheep. Measurements of mean volumetric tooth mineral density and total tooth volume were performed using quantitative computed tomography. Microcomputed tomography was used to measure total enamel volume, volumetric enamel mineral density, total dentin volume, and volumetric dentin mineral density. Maximum elastic strength and ultimate force of teeth were determined using 3-point bending and compression tests. Pearson correlation coefficients were determined between all investigated variables. Mutual dependence was observed between morphological and mechanical properties of the investigated teeth. The highest number of positive correlations of the investigated parameters was stated in first incisor indicating its superior predictive value of tooth quality and masticatory organ function in sheep. Positive correlations of the volumetric dentin mineral density in second premolar with final body weight may indicate predictive value of this parameter in relation with growth rate in sheep. Evaluation of deciduous tooth properties may prove helpful for breeding selection and further reproduction of sheep possessing favorable traits of teeth and better masticatory organ function, leading to improved performance and economic efficiency of the flock.

Simultaneous Quantification of Syringic Acid and Kaempferol in Extracts of Bergenia Species Using Validated High-Performance Thin-Layer Chromatographic-Densitometric Method.

PubMed

Srivastava, Nishi; Srivastava, Amit; Srivastava, Sharad; Rawat, Ajay Kumar Singh; Khan, Abdul Rahman

2016-03-01

A rapid, sensitive, selective and robust quantitative densitometric high-performance thin-layer chromatographic method was developed and validated for separation and quantification of syringic acid (SYA) and kaempferol (KML) in the hydrolyzed extracts of Bergenia ciliata and Bergenia stracheyi. The separation was performed on silica gel 60F254 high-performance thin-layer chromatography plates using toluene : ethyl acetate : formic acid (5 : 4: 1, v/v/v) as the mobile phase. The quantification of SYA and KML was carried out using a densitometric reflection/absorption mode at 290 nm. A dense spot of SYA and KML appeared on the developed plate at a retention factor value of 0.61 ± 0.02 and 0.70 ± 0.01. A precise and accurate quantification was performed using linear regression analysis by plotting the peak area vs concentration 100-600 ng/band (correlation coefficient: r = 0.997, regression coefficient: R(2) = 0.996) for SYA and 100-600 ng/band (correlation coefficient: r = 0.995, regression coefficient: R(2) = 0.991) for KML. The developed method was validated in terms of accuracy, recovery and inter- and intraday study as per International Conference on Harmonisation guidelines. The limit of detection and limit of quantification of SYA and KML were determined, respectively, as 91.63, 142.26 and 277.67, 431.09 ng. The statistical data analysis showed that the method is reproducible and selective for the estimation of SYA and KML in extracts of B. ciliata and B. stracheyi. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Morphometric, densitometric and mechanical properties of mandibular deciduous teeth in 5-month-old Polish Merino sheep

PubMed Central

2014-01-01

Background Caries, enamel hypoplasia, molar incisor hipomineralization, amylogenesis imperfecta, dentine dysplasia, hypophosphatasia and other dental disorders lead to tooth mineralization disturbances and structural abnormalities, decreasing masticatory organ functions. Dental disorders in sheep can lead to premature slaughter before they have attained final stage of their reproductive life and induce economic loss due to high flock replacement costs. Growth rate, health status and meat quality of sheep depends on tooth properties and quality determining in large extent efficiency of the masticatory apparatus and initial food break up. Considering lack of basic anatomical and physiological data on teeth properties in sheep, the aim of the study was to evaluate morphometric, densitometric and mechanical traits of deciduous mandibular incisor, canine and the second premolar obtained at the slaughter age of 5 months of life. Results The obtained results have shown the highest values of weight, total tooth volume, enamel volume and dentine volume in second premolar. Morphometric and mechanical parameters of incisors reached the highest values in first incisor and decreased gradually in second and third incisor, and in canine. Densitometric measurements have not revealed significant differences of the volumetric tooth mineral density in hard dental tissues between the investigated teeth. Conclusions In conclusion, proposed methodological approach is noninvasive since the deciduous teeth undergo physiological replacement with permanent teeth. Deciduous teeth can be easy collected for analyses from large animal population and may reflect mineral status and metabolism resulting from postnatal growth and development of the whole flock. In individual cases, evaluation of properties of deciduous teeth may serve for breeding selection and further reproduction of sheep possessing favorable traits of teeth and better masticatory system functions. PMID:24548814

Artificial neural network in breast lesions from fine-needle aspiration cytology smear.

PubMed

Subbaiah, R M; Dey, Pranab; Nijhawan, Raje

2014-03-01

Artificial neural networks (ANNs) are applied in engineering and certain medical fields. ANN has immense potential and is rarely been used in breast lesions. In this present study, we attempted to build up a complete robust back propagation ANN model based on cytomorphological data, morphometric data, nuclear densitometric data, and gray level co-occurrence matrix (GLCM) of ductal carcinoma and fibroadenomas of breast cases diagnosed on fine-needle aspiration cytology (FNAC). We selected 52 cases of fibroadenomas and 60 cases of infiltrating ductal carcinoma of breast diagnosed on FNAC by two cytologists. Essential cytological data was quantitated by two independent cytologists (SRM, PD). With the help of Image J software, nuclear morphomeric, densitometric, and GLCM features were measured in all the cases on hematoxylin and eosin-stained smears. With the available data, an ANN model was built up with the help of Neurointelligence software. The network was designed as 41-20-1 (41 input nodes, 20 hidden nodes, 1 output node). The network was trained by the online back propagation algorithm and 500 iterations were done. Learning was adjusted after every iteration. ANN model correctly identified all cases of fibroadenomas and infiltrating carcinomas in the test set. This is one of the first successful composite ANN models of breast carcinomas. This basic model can be used to diagnose the gray zone area of the breast lesions on FNAC. We assume that this model may have far-reaching implications in future. Copyright © 2013 Wiley Periodicals, Inc.

Effects of administration of four different doses of Escherichia coli phytase on femur properties of 16-week-old turkeys.

PubMed

Tatara, Marcin R; Krupski, Witold; Kozłowski, Krzysztof; Drażbo, Aleksandra; Jankowski, Jan

2015-03-18

The enzyme phytase is able to initiate the release of phosphates from phytic acid, making it available for absorption within gastrointestinal tract and following utilization. The aim of the study was to determine effects of Escherichia coli phytase administration on morphological, densitometric and mechanical properties of femur in 16-week-old turkeys. One-day-old BUT Big-6 males were assigned to six weight-matched groups. Turkeys receiving diet with standard phosphorus (P) and calcium (Ca) content belonged to the positive control group (Group I). Negative control group (Group II) consisted of birds fed diet with lowered P and Ca content. Turkeys belonging to the remaining groups have received the same diet as group II but enriched with graded levels of Escherichia coli phytase: 125 (Group III), 250 (Group IV), 500 (Group V) and 1000 (Group VI) FTU/kg. At the age of 112 days of life, the final body weights were determined and the turkeys were sacrificed to obtain right femur for analyses. Geometric and densitometric properties of femur were determined using quantitative computed tomography (QCT) technique, while mechanical evaluation was performed in three-point bending test. Phytase administration increased cross-sectional area, second moment of inertia, mean relative wall thickness, cortical bone mineral density and maximum elastic strength decreasing cortical bone area of femur (P < 0.05). Reduced dietary Ca and P content decreased final body weight of turkeys by 6.5% (P = 0.006). The most advantageous effects of Escherichia coli phytase administration on geometric, densitometric and mechanical properties of femur were observed in turkeys receiving 125 and 250 FTU/kg of the diet. Phytase administration at the dosages of 500 and 1000 FTU/kg of the diet improved the final body weight in turkeys. The results obtained in this study indicate a possible practical application of Escherichia coli phytase in turkey feeding to improve skeletal system properties and function.

Application of a validated stability-indicating densitometric thin-layer chromatographic method to stress degradation studies on moxifloxacin.

PubMed

Motwani, Sanjay K; Khar, Roop K; Ahmad, Farhan J; Chopra, Shruti; Kohli, K; Talegaonkar, S

2007-01-16

A simple, sensitive, selective, precise and stability-indicating high-performance thin-layer chromatographic (HPTLC) method for densitometric determination of moxifloxacin both as a bulk drug and from pharmaceutical formulation was developed and validated as per the International Conference on Harmonization (ICH) guidelines. The method employed TLC aluminium plates pre-coated with silica gel 60F-254 as the stationary phase and the mobile phase consisted of n-propanol-ethanol-6M ammonia solution (4:1:2, v/v/v). Densitometric analysis of moxifloxacin was carried out in the absorbance mode at 298 nm. Compact spots for moxifloxacin were found at R(f) value of 0.58+/-0.02. The linear regression analysis data for the calibration plots showed good linear relationship with r=0.9925 in the working concentration range of 100-800 ng spot(-1). The method was validated for precision, accuracy, ruggedness, robustness, specificity, recovery, limit of detection (LOD) and limit of quantitation (LOQ). The LOD and LOQ were 3.90 and 11.83 ng spot(-1), respectively. Drug was subjected to acid and alkali hydrolysis, oxidation, dry heat, wet heat treatment and photodegradation. All the peaks of degradation products were well resolved from the standard drug with significantly different R(f) values. Statistical analysis proves that the developed HPTLC method is reproducible and selective. As the method could effectively separate the drug from its degradation products, it can be employed as stability-indicating one. Moreover, the proposed HPTLC method was utilized to investigate the kinetics of the acidic and alkaline degradation processes at different temperatures. Arrhenius plot was constructed and apparent pseudo-first-order rate constant, half-life and activation energy were calculated. In addition the pH-rate profile for degradation of moxifloxacin in constant ionic strength buffer solutions within the pH range 1.2-10.8 was studied.

Development and validation of a reversed-phase high-performance thin-layer chromatography-densitometric method for determination of atorvastatin calcium in bulk drug and tablets.

PubMed

Shirkhedkar, Atul A; Surana, Sanjay J

2010-01-01

Atorvastatin calcium is a synthetic HMG-CoA reductase inhibitor that is used as a cholesterol-lowering agent. A simple, sensitive, selective, and precise RP-HPTLC-densitometric determination of atorvastatin calcium both as bulk drug and from pharmaceutical formulation was developed and validated according to International Conference on Harmonization guidelines. The method used aluminum sheets precoated with silica gel 60 RP18F254S as the stationary phase, and the mobile phase consisted of methanol-water (3.5 + 1.5, v/v). The system gave a compact band for atorvastatin calcium with an Rf value of 0.62 +/- 0.02. Densitometric quantification was carried out at 246 nm. The linear regression analysis data for the calibration plots showed a good linear relationship with r = 0.9992 in the working concentration range of 100-800 ng/band. The method was validated for precision, accuracy, ruggedness, robustness, specificity, recovery, LOD, and LOQ. The LOD and LOQ were 6 and 18 ng, respectively. The drug underwent hydrolysis when subjected to acidic conditions and was found to be stable under alkali, oxidation, dry heat, and photodegradation conditions. Statistical analysis proved that the developed RP-HPTLC-densitometry method is reproducible and selective and that it can be applied for identification and quantitative determination of atorvastatin calcium in bulk drug and tablet formulation.

Detection Progress of Selected Drugs in TLC

PubMed Central

Pyka, Alina

2014-01-01

This entry describes applications of known indicators and dyes as new visualizing reagents and various visualizing systems as well as photocatalytic reactions and bioautography method for the detection of bioactive compounds including drugs and compounds isolated from herbal extracts. Broadening index, detection index, characteristics of densitometric band, modified contrast index, limit of detection, densitometric visualizing index, and linearity range of detected compounds were used for the evaluation of visualizing effects of applied visualizing reagents. It was shown that visualizing effect depends on the chemical structure of the visualizing reagent, the structure of the substance detected, and the chromatographic adsorbent applied. The usefulness of densitometry to direct detection of some drugs was also shown. Quoted papers indicate the detection progress of selected drugs investigated by thin-layer chromatography (TLC). PMID:24551853

Skeletal Geometry and Indices of Bone Strength in Artistic Gymnasts

PubMed Central

Dowthwaite, Jodi N.; Scerpella, Tamara A.

2010-01-01

This review addresses bone geometry and indices of skeletal strength associated with exposure to gymnastic loading during growth. A brief background characterizes artistic gymnastics as a mechanical loading model and outlines densitometric techniques, skeletal outcomes and challenges in assessment of skeletal adaptation. The literature on bone geometric adaptation to gymnastic loading is sparse and consists of results for disparate skeletal sites, maturity phases, gender compositions and assessment methods, complicating synthesis of an overriding view. Furthermore, most studies assess only females, with little information on males and adults. Nonetheless, gymnastic loading during growth appears to yield significant enlargement of total and cortical bone geometry (+10 to 30%) and elevation of trabecular density (+20%) in the forearm, yielding elevated indices of skeletal strength (+20 to +50%). Other sites exhibit more moderate geometric and densitometric adaptations (5 to 15%). Mode of adaptation appears to be site-specific; some sites demonstrate marked periosteal and endosteal expansion, whereas other sites exhibit negligible or moderate periosteal expansion coupled with endocortical contraction. Further research is necessary to address sex-, maturity- and bone tissue-specific adaptation, as well as maintenance of benefits beyond loading cessation. PMID:19949278

Effects of developer depletion on image quality of Kodak Insight and Ektaspeed Plus films.

PubMed

Casanova, M S; Casanova, M L S; Haiter-Neto, F

2004-03-01

To evaluate the effect of processing solution depletion on the image quality of F-speed dental X-ray film (Insight), compared with Ektaspeed Plus. The films were exposed with a phantom and developed in manual and automatic conditions, in fresh and progressively depleted solutions. The comparison was based on densitometric analysis and subjective appraisal. The processing solution depletion presented a different behaviour depending on whether manual or automatic technique was used. The films were distinctly affected by depleted processing solutions. The developer depletion was faster in automatic than manual conditions. Insight film was more resistant than Ektaspeed Plus to the effects of processing solution depletion. In the present study there was agreement between the objective and subjective appraisals.

Estimation of yohimbine base in complex mixtures by quantitative HPTLC application.

PubMed

Adel-Kader, Maged Saad; Alwahebi, Naif Wahebi Hamadan; Alam, Prawez

2017-01-01

The indole alkaloid Yohimbine has been used for over two centuries in the treatment of erectly dysfunction. Several formulations containing yohimbine salts, yohimbe bark power or extract are marketed worldwide. Determination of the amount of yohimbine in such formulation is a challenging task due to their complex nature. Extraction followed by acid-base purification resulted in a relatively pure alkaloids containing fractions. The exact amounts of yohimbine free base in different formulations were determined by densitometric HPTLC validated methods using silica gel TLC plates. Standard curve for yohimbine was generated using yohimbine hydrochloride subjected to the same acid-base treatment as the used samples. All formulations found to contain yohimbine though some with less concentration than the labeled amount.

Stability-Indicating TLC-Densitometric Assay for Methyltestosterone and Quantum Chemical Calculations.

PubMed

Musharraf, Syed Ghulam; Ul Arfeen, Qamar; Ul Haq, Faraz; Khatoon, Aliya; Azher Ali, Rahat

2017-10-01

Methyltestosterone is a synthetic testosterone derivative commonly used for the treatment of testosterone deficiency in males and one the anabolic steroids whose use is banned by World Anti-Doping Agency (WADA). This study presents a simple, cost-effective and rapid stability-indicating assay for densitometric quantification of methyltestosterone in pharmaceutical formulation. The developed method employed pre-coated TLC plates with mobile phase hexane:acetone (6.5:3.5 v/v). Limit of detection and limit of quantitation were found to be 2.06 and 6.24 ng/spot, respectively. Stress degradation study of methyltestosterone was conducted by applying various stress conditions such as hydrolysis under acidic, basic and neutral conditions, heating in anhydrous conditions and exposure to light. Methyltestosterone was found to be susceptible to photodegradation, acidic and basic hydrolysis. Degraded products were well resolved with significantly different Rf values. Acid degraded product was identified as 17,17-dimethyl-18-norandrosta-4,13(14)-dien-3-one through spectroscopic methods. The reactivity of methyltestosterone under applied stress conditions was also explained by quantum chemical calculations. The developed method is found to be repeatable, selective and accurate for quantification of methyltestosterone and can be employed for routine analysis. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

First derivative ratio spectrophotometric, HPTLC-densitometric, and HPLC determination of nicergoline in presence of its hydrolysis-induced degradation product.

PubMed

Ahmad, Abdel Kader S; Kawy, M Abdel; Nebsen, M

2002-10-15

Three methods are presented for the determination of Nicergoline in presence of its hydrolysis-induced degradation product. The first method was based on measurement of the first derivative of ratio spectra amplitude of Nicergoline at 291 nm. The second method was based on separation of Nicergoline from its degradation product followed by densitometric measurement of the spots at 287 nm. The separation was carried out on HPTLC silica gel F(254) plates, using methanol-ethyl acetate-glacial acetic acid (5:7:3, v/v/v) as mobile phase. The third method was based on high performance liquid chromatographic (HPLC) separation and determination of Nicergoline from its degradation product on a reversed phase, nucloesil C(18) column using a mobile phase of methanol-water-glacial acetic acid (80:20:0.1, v/v/v) with UV detection at 280 nm. Chlorpromazine hydrochloride was used as internal standard. Laboratory prepared mixtures containing different percentages of the degradation product were analysed by the proposed methods and satisfactory results were obtained. These methods have been successfully applied to the analysis of Nicergoline in Sermion tablets. The validities of these methods were ascertained by applying standard addition technique, the mean percentage recovery +/- R.S.D.% was found to be 99.47 +/- 0.752, 100.01 +/- 0.940, 99.75 +/- 0.740 for the first derivative of ratio spectra method, the HPTLC method and the HPLC method, respectively. The proposed methods were statistically compared with the manufacturer's HPLC method of analysis of Nicergoline and no significant difference was found with respect to both precision and accuracy. They have the advantage of being stability indicating. Therefore, they can be used for routine analysis of the drug in quality control laboratories. Copyright 2002 Elsevier Science B.V.

Soil, water, and vegetation conditions in south Texas

NASA Technical Reports Server (NTRS)

Wiegand, C. L.; Gausman, H. W.; Leamer, R. W.; Richardson, A. J.; Everitt, J. H.; Gerbermann, A. H. (Principal Investigator)

1976-01-01

The author has identified the following significant results. Field spectral measurements and laboratory densitometric measurements showed that tree canopy reflectance differences among the Marrs, Redblush, and Valencia varieties in the visible spectral region were due to their different leaf chlorophyll concentrations. Field measurements of visible light reflectance were directly related to the tonal responses on infrared color photos of the varietal tree canopies. Consequently, densitometric measurements of the foliage on the infrared color transparency with red-filtered light successfully discriminated among the three varieties. Reflectance measurements with a field spectroradiometer on nine dates the growing season of two wheat varieties, Milam and Penjamo, documented their spectra over the 0.45 to 2.50 micron wavelength interval associated with plant cover and physiological development. An image analyzer system was used to optically planimeter the percentage of soil background, vegetation and shadow in the vertical photographs taken within the FOV of the spectroradiometer on each measurement date.

Validated stability-indicating densitometric thin-layer chromatography: application to stress degradation studies of minocycline.

PubMed

Jain, Nilu; Jain, Gaurav Kumar; Ahmad, Farhan Jalees; Khar, Roop Krishen

2007-09-19

A simple, stability-indicating high-performance thin-layer liquid chromatographic (HPTLC) method for analysis of minocycline was developed and validated. The densitometric analysis was carried out at 345 nm using methanol-acetonitrile-isopropyl alcohol-water (5:4:0.5:0.5, v/v/v/v) as mobile phase. The method employed TLC aluminium plates pre-coated with silica gel 60F-254 as the stationary phase. To achieve good result, plates were sprayed with a 10% (w/v) solution of disodium ethylene diaminetetraacetic acid (EDTA), the pH of which was adjusted to 9.0. Compact spots of minocycline were found at R(f) = 0.30+/-0.02. For proposed procedure, linearity (r = 0.9997), limit of detection (3.7 ng spot(-1)), recovery (99.23-100.16%), and precision (% R.S.D. < or = 0.364) was found to be satisfactory. The drug undergoes acidic and basic degradation, oxidation and photodegradation. All the peaks of degradation products were well resolved from the pure drug with significantly different R(f) values. The acidic and alkaline degradation kinetics of minocycline, evaluated using this method, is found to be of first order.

RP-LC and TLC Densitometric Determination of Paracetamol and Pamabrom in Presence of Hazardous Impurity of Paracetamol and Application to Pharmaceuticals

PubMed Central

El-Houssini, Ola Mohamed

2013-01-01

Two simple, accurate and reproducible methods were developed and validated for the simultaneous determination of paracetamol (PARA) and pamabrom (PAMB) in pure form and in tablets. The first method was based on reserved-phase high-performance liquid chromatography, on a Thermo Hypersil ODS column using methanol:0.01 M sodium hexane sulfonate:formic acid (67.5:212.5:1 v/v/v) as the mobile phase. The flow rate was 2 mL/min and the column temperature was adjusted to 35 °C. Quantification was achieved with UV detection at 277 nm over concentration range of 100–600 and 4–24 μg/mL, with mean percentage recoveries were found to be 99.90 ± 0.586 and 99.26 ± 0.901 for PARA and PAMB, respectively. The second method was based on thin-layer chromatography separation of PARA and PAMB followed by densitometric measurement of the spots at 254 nm and 277 nm for PARA and PAMB respectively. Separation was carried out on aluminum sheet of silica gel 60F254 using dichloromethane:methanol:glacial acetic acid (7.5:1:0.5 v/v/v) as the mobile phase over concentration range of 1–10 and 0.32–3.20 μg per spot, with mean percentage recovery of 100.52 ± 1.332 and 99.71 ± 1.478 for PARA and PAMB, respectively. The methods retained their accuracy in presence of up to 50% of P-aminophenol and could be successfully applied in tablets. PMID:24046511

High performance liquid chromatographic and thin layer densitometric methods for the determination of risperidone in the presence of its degradation products in bulk powder and in tablets.

PubMed

El-Sherif, Zeinab A; El-Zeany, Badr; El-Houssini, Ola M

2005-01-04

Two reproducible stability indicating methods were developed for the determination of risperidone (RISP) in presence of its degradation products in pure form and in tablets. The first method was based on reversed phase high performance liquid chromatography (HPLC), on Lichrosorb RP C 18 column (250 mm i.d., 4 mm, 10 microm), using methanol:0.05 M potassium dihydrogen phosphate pH 7 (65:35 (v/v)) as the mobile phase at a flow rate of 1 ml min(-1) at ambient temperature. Quantification was achieved with UV detection at 280 nm over a concentration range of 25-500 microg ml(-1) with mean percentage recovery of 99.87 +/- 1.049. The method retained its accuracy in the presence of up to 90% of RISP degradation products. The second method was based on TLC separation of RISP from its degradation products followed by densitometric measurement of the intact drug spot at 280 nm. The separation was carried out on aluminum sheet of silica gel 60F254 using acetonitrile:methanol:propanol:triethanolamine (8.5:1.2:0.6:0.2 (v/v/v/v)), as the mobile phase, over a concentration range of 2-10 microg per spot and mean percentage recovery of 100.1 +/- 1.18. The two methods were simple, precise, sensitive and could be successfully applied for the determination of pure, laboratory prepared mixtures and tablets. The results obtained were compared with the manufacturer's method.

The use of ERTS imagery for lake classification. [turbidity due to phytoplankton

NASA Technical Reports Server (NTRS)

Scarpace, F. L.; Wade, R. E.; Fisher, L. T.

1975-01-01

The feasibility of using photographic representations of the ERTS imagery to classify lakes in the State of Wisconsin as to their trophic level was studied. Densitometric readings in band 5 of ERTS 70 mm imagery were taken for all the lakes in Wisconsin greater than 100 acres (approximately 1000 lakes). An algorithm has been developed from ground truth measurements to predict from satellite imagery an indicator of trophic status.

The calibration of photographic and spectroscopic films. A densitometric analysis of IIaO film flown aboard the space shuttle transportation system STS3, STS8, and STS7

NASA Technical Reports Server (NTRS)

Hammond, Ernest C., Jr.

1987-01-01

The results of these studies have implications for the utilization of the IIaO spectroscopic film on the future shuttle and space lab missions. These responses to standard photonic energy sources will have immediate application for the uneven responses of the film photographing a star field in a terrestrial or extraterrestrial environment with associated digital imaging equipment.

A new high-performance thin-layer chromatographic method for determining bile salt hydrolase activity.

PubMed

Rohawi, Nur Syakila; Ramasamy, Kalavathy; Agatonovic-Kustrin, Snezana; Lim, Siong Meng

2018-06-05

A quantitative assay using high-performance thin-layer chromatography (HPTLC) was developed to investigate bile salt hydrolase (BSH) activity in Pediococcus pentosaceus LAB6 and Lactobacillus plantarum LAB12 probiotic bacteria isolated from Malaysian fermented food. Lactic acid bacteria (LAB) were cultured in de Man Rogosa and Sharpe (MRS) broth containing 1 mmol/L of sodium-based glyco- and tauro-conjugated bile salts for 24 h. The cultures were centrifuged and the resultant cell free supernatant was subjected to chromatographic separation on a HPTLC plate. Conjugated bile salts were quantified by densitometric scans at 550 nm and results were compared to digital image analysis of chromatographic plates after derivatisation with anisaldehyde/sulfuric acid. Standard curves for bile salts determination with both methods show good linearity with high coefficient of determination (R 2 ) between 0.97 and 0.99. Method validation indicates good sensitivity with low relative standard deviation (RSD) (<10%), low limits of detection (LOD) of 0.4 versus 0.2 μg and limit of quantification (LOQ) of 1.4 versus 0.7 μg, for densitometric vs digital image analysis method, respectively. The bile salt hydrolase activity was found to be higher against glyco- than tauro-conjugated bile salts (LAB6; 100% vs >38%: LAB12; 100% vs >75%). The present findings strongly show that quantitative analysis via digitally-enhanced HPTLC offers a rapid quantitative analysis for deconjugation of bile salts by probiotics. Copyright © 2018. Published by Elsevier B.V.

Larval x-ray irradiation influences protein expression in pupae of the Oriental fruit fly, Bactrocera Dorsalis

USDA-ARS?s Scientific Manuscript database

Third instar larvae were exposed to X-ray treatment of the Oriental fruit fly, Bactrocera dorsalis. Irradiated pupae were collected daily. Biological performance parameters of pupae and adults of larvae treated with X-ray irradiation were evaluated. Standard proteomics procedures such as densitometr...

Estimation of L-dopa from Mucuna pruriens LINN and formulations containing M. pruriens by HPTLC method.

PubMed

Modi, Ketan Pravinbhai; Patel, Natvarlal Manilal; Goyal, Ramesh Kishorilal

2008-03-01

A selective, precise, and accurate high-performance thin-layer chromatographic (HPTLC) method has been developed for the analysis of L-dopa in Mucuna pruriens seed extract and its formulations. The method involves densitometric evaluation of L-dopa after resolving it by HPTLC on silica gel plates with n-butanol-acetic acid-water (4.0+1.0+1.0, v/v) as the mobile phase. Densitometric analysis of L-dopa was carried out in the absorbance mode at 280 nm. The relationship between the concentration of L-dopa and corresponding peak areas was found to be linear in the range of 100 to 1200 ng/spot. The method was validated for precision (inter and intraday), repeatability, and accuracy. Mean recovery was 100.30%. The relative standard deviation (RSD) values of the precision were found to be in the range 0.64-1.52%. In conclusion, the proposed TLC method was found to be precise, specific and accurate and can be used for identification and quantitative determination of L-dopa in herbal extract and its formulations.

A validated densitometric method for analysis of atorvastatin calcium and metoprolol tartarate as bulk drugs and in combined capsule dosage forms.

PubMed

Patole, Sm; Khodke, As; Potale, Lv; Damle, Mc

2011-01-01

A simple, accurate and precise high-performance thin-layer chromatographic method has been developed for the estimation of Atorvastatin Calcium and Metoprolol Tartarate simultaneously from a capsule dosage form. The method employed Silica gel 60F (254s)precoated plates as stationary phase and a mixture of Chloroform: Methanol: Glacial acetic acid (dil.) :: (9:1.5:0.2 ml %v/v) as mobile phase. Densitometric scanning was performed at 220 nm using Camag TLC scanner 3. The method was linear in the drug concentrations' range of 500 to 2500 ng/spot for Atorvastatin Calcium, also for Metoprolol Tartarate with correlation coefficient of 0.984 for Atorvastatin Calcium and 0.995 for Metoprolol Tartarate respectively. The retention factor for Atorvastatin Calcium was 0.45 ± 0.04 and for Metoprolol Tartarate was 0.25 ± 0.02. The method was validated as per ICH (International Conference on Harmonisation) Guidelines, proving its utility in estimation of Atorvastatin Calcium and Metoprolol Tartarate in combined dosage form.

The application of satellite data in monitoring strip mines

NASA Technical Reports Server (NTRS)

Sharber, L. A.; Shahrokhi, F.

1977-01-01

Strip mines in the New River Drainage Basin of Tennessee were studied through use of Landsat-1 imagery and aircraft photography. A multilevel analysis, involving conventional photo interpretation techniques, densitometric methods, multispectral analysis and statistical testing was applied to the data. The Landsat imagery proved adequate for monitoring large-scale change resulting from active mining and land-reclamation projects. However, the spatial resolution of the satellite imagery rendered it inadequate for assessment of many smaller strip mines, in the region which may be as small as a few hectares.

MicroCT-based phenomics in the zebrafish skeleton reveals virtues of deep phenotyping in a distributed organ system.

PubMed

Hur, Matthew; Gistelinck, Charlotte A; Huber, Philippe; Lee, Jane; Thompson, Marjorie H; Monstad-Rios, Adrian T; Watson, Claire J; McMenamin, Sarah K; Willaert, Andy; Parichy, David M; Coucke, Paul; Kwon, Ronald Y

2017-09-08

Phenomics, which ideally involves in-depth phenotyping at the whole-organism scale, may enhance our functional understanding of genetic variation. Here, we demonstrate methods to profile hundreds of phenotypic measures comprised of morphological and densitometric traits at a large number of sites within the axial skeleton of adult zebrafish. We show the potential for vertebral patterns to confer heightened sensitivity, with similar specificity, in discriminating mutant populations compared to analyzing individual vertebrae in isolation. We identify phenotypes associated with human brittle bone disease and thyroid stimulating hormone receptor hyperactivity. Finally, we develop allometric models and show their potential to aid in the discrimination of mutant phenotypes masked by alterations in growth. Our studies demonstrate virtues of deep phenotyping in a spatially distributed organ system. Analyzing phenotypic patterns may increase productivity in genetic screens, and facilitate the study of genetic variants associated with smaller effect sizes, such as those that underlie complex diseases.

Densitometric evaluation of Soludent and GBX developers.

PubMed

Patel, J R

1985-01-01

A quick-developing solution (Soludent) and a new developer (Kodak GBX) were compared with a standard x-ray liquid developer. Of the three solutions evaluated, Kodak GBX solution produced slightly greater useful densities in the radiograph at all temperatures evaluated. The rapid-developing solution produced acceptable radiographs in 80% less time, with only slightly higher film fog.

Videodensitometric Methods for Cardiac Output Measurements

NASA Astrophysics Data System (ADS)

Mischi, Massimo; Kalker, Ton; Korsten, Erik

2003-12-01

Cardiac output is often measured by indicator dilution techniques, usually based on dye or cold saline injections. Developments of more stable ultrasound contrast agents (UCA) are leading to new noninvasive indicator dilution methods. However, several problems concerning the interpretation of dilution curves as detected by ultrasound transducers have arisen. This paper presents a method for blood flow measurements based on UCA dilution. Dilution curves are determined by real-time densitometric analysis of the video output of an ultrasound scanner and are automatically fitted by the Local Density Random Walk model. A new fitting algorithm based on multiple linear regression is developed. Calibration, that is, the relation between videodensity and UCA concentration, is modelled by in vitro experimentation. The flow measurement system is validated by in vitro perfusion of SonoVue contrast agent. The results show an accurate dilution curve fit and flow estimation with determination coefficient larger than 0.95 and 0.99, respectively.

Validated Chromatographic Methods for the Analysis of Two Binary Mixtures Containing Pyridoxine Hydrochloride.

PubMed

Habib, Neven M; Abdelrahman, Maha M; Abdelwhab, Nada S; Ali, Nourudin W

2017-03-01

Accurate and precise TLC-densitometric and HPLC-diode-array detector (DAD) methods have been developed and validated to resolve two binary mixtures containing pyridoxine hydrochloride (PYH) with either cyclizine hydrochloride (CYH) or meclizine hydrochloride (MEH). In the developed TLC-densitometric method, chromatographic separation of the three studied drugs was carried out on silica gel 60 F254 plates using a developing system containing methylene chloride + acetone + methanol (7 + 1 + 0.5, v/v/v) scanning separated bands at 220 nm. Beer-Lambert law was obeyed in the ranges of 0.2-5, 0.2-4, and 0.2-4 µg/band for PYH, CYH, and MEH, respectively. On the other hand, the developed HPLC-DAD method depended on chromatographic separation on a Zorbax Eclipse C18 column using methanol-KH2PO4 (0.05 M; 90 + 10, v/v; pH 5, with H3PO4 and KOH) as the mobile phase, a flow rate of 1 mL/min, and UV scanning at 220 nm. A linear relationship was obtained between the integrated peak area and the concentration in the ranges of 10-50, 10-50, and 7-50 µg/mL for PYH, CYH, and MEH, respectively. The proposed methods were successfully applied for the determination of the cited drugs in their pharmaceutical formulations. Statistical comparison with the reported methods using Student's t- and F-tests found there were no significant differences between the proposed and reported methods for accuracy and precision.

Simultaneous Determination of Acetylsalicylic Acid, Hydrochlorothiazide, Enalapril, and Atorvastatin in a Polypill-Based Quaternary Mixture by TLC.

PubMed

Maślanka, Anna; Stolarczyk, Mariusz; Apola, Anna; Kwiecień, Anna; Hubicka, Urszula; Opoka, Włodzimierz

2018-05-01

A new chromatographic-densitometric method has been developed for the qualitative and quantitative determination of the active ingredients in a simulated mixture corresponding to the PolyIran polypill, composed of acetylsalicylic acid, hydrochlorothiazide (HCT), enalapril (ENA), and atorvastatin (ATR), whose efficacy in the treatment and prevention of cardiovascular disease has been documented in clinical trials. Chromatographic separation was performed using TLC silica gel 60 plates with fluorescent indicator F254 as the stationary phase and a mixture of n-hexane-ethyl acetate-methanol-water-acetic acid (8.4 + 8 + 3 + 0.4 + 0.2, v/v/v/v/v) as the mobile phase. Densitometric measurements were carried out at λ = 210 nm when determining ENA and at λ = 265 nm in the case of the other drugs. Peaks of examined substances were well separated in the recorded chromatograms, enabling the evaluation of the results in terms of both qualitative and quantitative analysis. The method was specific for the analyzed components and was characterized by high sensitivity. The LOD was between 0.043 and 0.331 μg/spot, and LOQ was between 0.100 and 0.942 μg/spot. Recovery was in the range of 97.02-101.34%. The linearity range was broad and ranged from 0.600 to 6.000 μg/spot for acetylsalicylic acid, from 0.058 to 1.102 μg/spot for HCT, from 0.505 to 6.560 μg/spot for ENA, and from 0.100 to 1.000 μg/spot for ATR. The method was characterized by good precision, with RSD values that ranged from 0.10 to 2.26%.

Dosage variability of topical ocular hypotensive products: a densitometric assessment.

PubMed

Gaynes, Bruce I; Singa, Ramesh M; Cao, Ying

2009-02-01

To ascertain consequence of variability in drop volume obtained from multiuse topical ocular hypotensive products in terms of uniformity of product dosage. Densitometric assessment of drop volume dispensed from 2 alternative bottle positions. All except one product demonstrated a statistically significant difference in drop volume when administered at either a 45-degree or 90-degree bottle angle (Student t test, P<0.001). Product-specific drop volume ranged from a nadir of 22.36 microL to a high of 53.54 microL depending on bottle angle of administration. Deviation in drop dose was directly proportional to variability in drop volume. Variability in per drop dosage was conspicuous among products with a coefficient of variation from 1.49% to 15.91%. In accordance with drop volume, all products demonstrated a statistically significant difference in drop dose at 45-degree versus 90-degree administration angles. Drop volume was found unrelated to drop uniformity (Spearman r=0.01987 and P=0.9463). Variability and lack of uniformity in drop dosage is clearly evident among select ocular hypotensive products and is related to angle of drop administration. Erratic dosage of topical ocular hypotensive therapy may contribute in part to therapeutic failure and/or toxicity.

Densitometric HPTLC analysis of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements, teas and commercial creams.

PubMed

Alam, Prawez

2013-08-01

To develop and validate a simple, accurate HPTLC method for the analysis of 8-gingerol and to determine the quantity of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements, teas and commercial creams. The analysis was performed on 10×20 cm aluminium-backed plates coated with 0.2 mm layers of silica gel 60 F254 (E-Merck, Germany) with n-hexane: ethyl acetate 60: 40 (v/v) as mobile phase. Camag TLC Scanner III was used for the UV densitometric scanning at 569. This system was found to give a compact spot of 8-gingerol at retention factor (Rf) value of (0.39±0.04) and linearity was found in the ranges 50-500 ng/spot (r (2)=0.9987). Limit of detection (12.76 ng/spot), limit of quantification (26.32 ng/spot), accuracy (less than 2 %) and recovery (ranging from 98.22-99.20) were found satisfactory. The HPTLC method developed for quantification of 8-gingerol was found to be simple, accurate, reproducible, sensitive and is applicable to the analysis of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements, teas and commercial creams.

Correlation of DNA content and nucleomorphometric features with World Health Organization grading of meningiomas.

PubMed

Grunewald, J P; Röhl, F W; Kirches, E; Dietzmann, K

1998-02-01

Many studies dealing with extracranial cancer showed a strong correlation of DNA ploidy to a poor clinical outcome, recurrence, or malignancy. In brain tumors, analysis of DNA content did not always provided significant diagnostic information. In this study, DNA density and karyometric parameters of 50 meningiomas (26 Grade I, 10 Grade II, 14 Grade III) were quantitatively evaluated by digital cell image analyses of Feulgen-stained nuclei. In particular, the densitometric parameter SEXT, which describes nuclear DNA content, as well as the morphometric values LENG (a computer-assisted measurement of nuclear circumference), AREA (a computer-assisted measurement of nuclear area), FCON (a parameter that describes nuclear roundness), and CONC (a describing nuclear contour), evaluated with the software IMAGE C, were correlated to World Health Organization (WHO) grading using univariate and multivariate methods. AREA and LENG values showed significant differences between tumors of Grades I and III. FCON values were unable to distinguish WHO Grade III from Grade I/II but were useful in clearly separating Grade II from Grade I tumors. CONC values detected differences between WHO Grades II and I/III tumors but not between the latter. SEXT values clearly distinguished Grade III from Grade I/II tumors. The 1c, 2c, 2.5c, and 5c exceeding rates showed no predictive values. Only the 6c exceeding rate showed a significant difference between Grades I and III. These results outline the characteristic features of the atypical (Grade II) meningiomas, which make them a recognizable tumor entity distinct from benign and anaplastic meningiomas. The combination of DNA densitometric and morphometric findings seems to be a powerful addition to the histopathologic classification of meningiomas, as suggested by the WHO.

Quantitative estimation of pulegone in Mentha longifolia growing in Saudi Arabia. Is it safe to use?

PubMed

Alam, Prawez; Saleh, Mahmoud Fayez; Abdel-Kader, Maged Saad

2016-03-01

Our TLC study of the volatile oil isolated from Mentha longifolia showed a major UV active spot with higher Rf value than menthol. Based on the fact that the components of the oil from same plant differ quantitatively due to environmental conditions, the major spot was isolated using different chromatographic techniques and identified by spectroscopic means as pulegone. The presence of pulegone in M. longifolia, a plant widely used in Saudi Arabia, raised a hot debate due to its known toxicity. The Scientific Committee on Food, Health & Consumer Protection Directorate General, European Commission set a limit for the presence of pulegone in foodstuffs and beverages. In this paper we attempted to determine the exact amount of pulegone in different extracts, volatile oil as well as tea flavoured with M. longifolia (Habak) by densitometric HPTLC validated methods using normal phase (Method I) and reverse phase (Method II) TLC plates. The study indicated that the style of use of Habak in Saudi Arabia resulted in much less amount of pulegone than the allowed limit.

Management of natural resources through automatic cartographic inventory

NASA Technical Reports Server (NTRS)

Rey, P. A.; Gourinard, Y.; Cambou, F. (Principal Investigator)

1974-01-01

The author has identified the following significant results. Significant correspondence codes relating ERTS imagery to ground truth from vegetation and geology maps have been established. The use of color equidensity and color composite methods for selecting zones of equal densitometric value on ERTS imagery was perfected. Primary interest of temporal color composite is stressed. A chain of transfer operations from ERTS imagery to the automatic mapping of natural resources was developed.

An evaluation of space acquired data as a tool for wildlife management in Alaska

NASA Technical Reports Server (NTRS)

Vantries, B. J. (Principal Investigator)

1973-01-01

The author has identified the following significant results. Interpretation of ERTS-1 imagery by color-coded densitometric displays and digital processes data verified that with adequate quadrat in situ sampling ERTS-1 data could be extrapolated to describe accurately the vegetative characteristics of analogous sites, and that surface acres of water for waterfowl production were obtainable for ponds a minimum of 5 acres in size.

Bone Geometry as a Predictor of Tissue Fragility and Stress Fracture Risk

DTIC Science & Technology

2006-10-01

WidthML/2) directions. Moment of inertia and section modulus were assessed because these geometric measures are related to the bending and...mandible: Thomas; 1963. Ferretti JL, Capozza RF, Mondelo N, Zanchetta JR. Interrelationships between densitometric, geometric , and mechanical...fractures. Am J Sports Med, 28:245-251, 2000. Miller GJ, Purkey WW, Jr. The geometric properties of paired human tibiae. J Biomech, 13:1-8, 1980

Uni-dimensional double development HPTLC-densitometry method for simultaneous analysis of mangiferin and lupeol content in mango (Mangifera indica) pulp and peel during storage.

PubMed

Jyotshna; Srivastava, Pooja; Killadi, Bharti; Shanker, Karuna

2015-06-01

Mango (Mangifera indica) fruit is one of the important commercial fruit crops of India. Similar to other tropical fruits it is also highly perishable in nature. During storage/ripening, changes in its physico-chemical quality parameters viz. firmness, titrable acidity, total soluble solid content (TSSC), carotenoids content, and other biochemicals are inevitable. A uni-dimensional double-development high-performance thin-layer chromatography (UDDD-HPTLC) method was developed for the real-time monitoring of mangiferin and lupeol in mango pulp and peel during storage. The quantitative determination of both compounds of different classes was achieved by densitometric HPTLC method. Silica gel 60F254 HPTLC plates and two solvent systems viz. toluene/EtOAC/MeOH and EtOAC/MeOH, respectively were used for optimum separation and selective evaluation. Densitometric quantitation of mangiferin was performed at 390nm, while lupeol at 610nm after post chromatographic derivatization. Validated method was used to real-time monitoring of mangiferin and lupeol content during storage in four Indian cultivars, e.g. Bombay green (Bgreen), Dashehari, Langra, and Chausa. Significant correlations (p<0.05) between of acidity and TSSC with mangiferin and lupeol in pulp and peel during storage were also observed. Copyright © 2014 Elsevier Ltd. All rights reserved.

Densitometric HPTLC analysis of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements, teas and commercial creams

PubMed Central

Alam, Prawez

2013-01-01

Objective To develop and validate a simple, accurate HPTLC method for the analysis of 8-gingerol and to determine the quantity of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements, teas and commercial creams. Methods The analysis was performed on 10×20 cm aluminium-backed plates coated with 0.2 mm layers of silica gel 60 F254 (E-Merck, Germany) with n-hexane: ethyl acetate 60: 40 (v/v) as mobile phase. Camag TLC Scanner III was used for the UV densitometric scanning at 569. Results This system was found to give a compact spot of 8-gingerol at retention factor (Rf) value of (0.39±0.04) and linearity was found in the ranges 50-500 ng/spot (r2=0.9987). Limit of detection (12.76 ng/spot), limit of quantification (26.32 ng/spot), accuracy (less than 2 %) and recovery (ranging from 98.22-99.20) were found satisfactory. Conclusions The HPTLC method developed for quantification of 8-gingerol was found to be simple, accurate, reproducible, sensitive and is applicable to the analysis of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements, teas and commercial creams. PMID:23905021

Multiplex titration RT-PCR: rapid determination of gene expression patterns for a large number of genes

NASA Technical Reports Server (NTRS)

Nebenfuhr, A.; Lomax, T. L.

1998-01-01

We have developed an improved method for determination of gene expression levels with RT-PCR. The procedure is rapid and does not require extensive optimization or densitometric analysis. Since the detection of individual transcripts is PCR-based, small amounts of tissue samples are sufficient for the analysis of expression patterns in large gene families. Using this method, we were able to rapidly screen nine members of the Aux/IAA family of auxin-responsive genes and identify those genes which vary in message abundance in a tissue- and light-specific manner. While not offering the accuracy of conventional semi-quantitative or competitive RT-PCR, our method allows quick screening of large numbers of genes in a wide range of RNA samples with just a thermal cycler and standard gel analysis equipment.

The influence of center of rotation on the assessment of trabecular bone densitometric and structural properties.

PubMed

Sheng, Zhi-Feng; Dai, Ru-Chun; Wu, Xian-Ping; Ma, Yu-Lin; Xu, Kang; Zhang, Yu-Hai; Jiang, Ye-Bin; Liao, Er-Yuan

2008-12-01

The center of rotation is a physical location in the microCT scanner, defined by the axis of rotation of the sample stage. This physical location is always well defined during calibration of the instrument and fitted by an appropriate algorithm. However, in real images of limited contrast and with X-ray photon noise, this algorithm exhibits poorer precision and the optimum center of rotation cannot be always acquired. Thus, adjustment by operator is necessary to determine whether the center of rotation was correct, in order that the structural information of the sample can be correctly interpreted. In this paper, the effect of center of rotation on the assessment of densitometric and structural properties of trabecular bone was firstly evaluated. Twenty female Sprague-Dawley rats of 7-month-old were randomly assigned to ovariectomized (OVX) and SHAM-operated (SHAM) groups. The left tibiae were harvested at 3 weeks postoperatively. High resolution microCT was used to identify the densitometric and microstructural properties of trabeculae in the proximal ends of tibia. After CT scanning, the best artificial center of rotation for each scan was obtained. Bone parameters analyses were performed on the centers at different places away from the best artificial center of +/-0.2, +/-0.5, +/-1.0, +/-1.5, and +/-2.0 pixels, respectively. The general linear model (GLM) repeated measures procedure was used to investigate the difference in the parameters between the two groups (OVX vs. SHAM) and the possible effects of center displacements. A significant difference between OVX and SHAM groups was found in all parameters (p < 0.05) except Tb.Th, DA, and BS/BV. TBMD, DA, BS/BV, and Conn.D were decreased while BV/TV and Tb.Th were increased with the center deflection. Variations of these parameters were acceptable when the displacements were limited within +/-1.5 pixels for tBMD, BV/TV, DA, and Conn.D, and +/-1.0 pixels for Tb.Th and BS/BV. These changes were similar in both OVX and SHAM groups. The changing curves of bone parameters vs. centers could be well fitted by quadric regression models, by which the real center could be acquired, and thus the precision of microCT analysis would be improved. There were some inevitable differences between the best artificial and real centers.

Protein changes in leaf-sheath pulvini of barley (hordeum) induced by gravistimulation

NASA Technical Reports Server (NTRS)

Kaufman, P. B.; Song, I.

1984-01-01

Sodium dodecyl sulfate polyacrylamade gel electrophoresis (SDS-PAGE) pattern of salt soluble proteins elicite by gravistimulation were shown in the top and the bottom halves of the gravistimulated pulvini as follows: at least five proteins were increased in the tissues derived from the bottom halves of the pulvini in the approximate molecular weight range of 91, 57, 50, 22, 17 kilodatons. SDS densitometric scans indicated that the two of them are probably cellulase and invertase.

Thin layer chromatography-densitometric determination of some non-sedating antihistamines in combination with pseudoephedrine or acetaminophen in synthetic mixtures and in pharmaceutical formulations.

PubMed

El-Kommos, Michael E; El-Gizawy, Samia M; Atia, Noha N; Hosny, Noha M

2014-03-01

The combination of certain non-sedating antihistamines (NSA) such as fexofenadine (FXD), ketotifen (KET) and loratadine (LOR) with pseudoephedrine (PSE) or acetaminophen (ACE) is widely used in the treatment of allergic rhinitis, conjunctivitis and chronic urticaria. A rapid, simple, selective and precise densitometric method was developed and validated for simultaneous estimation of six synthetic binary mixtures and their pharmaceutical dosage forms. The method employed thin layer chromatography aluminum plates precoated with silica gel G 60 F254 as the stationary phase. The mobile phases chosen for development gave compact bands for the mixtures FXD-PSE (I), KET-PSE (II), LOR-PSE (III), FXD-ACE (IV), KET-ACE (V) and LOR-ACE (VI) [Retardation factor (Rf ) values were (0.20, 0.32), (0.69, 0.34), (0.79, 0.13), (0.36, 0.70), (0.51, 0.30) and (0.76, 0.26), respectively]. Spectrodensitometric scanning integration was performed at 217, 218, 218, 233, 272 and 251 nm for the mixtures I-VI, respectively. The linear regression data for the calibration plots showed an excellent linear relationship. The method was validated for precision, accuracy, robustness and recovery. Limits of detection and quantitation were calculated. Statistical analysis proved that the method is reproducible and selective for the simultaneous estimation of these binary mixtures. Copyright © 2013 John Wiley & Sons, Ltd.

Quantitative measurement of protein digestion in simulated gastric fluid.

PubMed

Herman, Rod A; Korjagin, Valerie A; Schafer, Barry W

2005-04-01

The digestibility of novel proteins in simulated gastric fluid is considered to be an indicator of reduced risk of allergenic potential in food, and estimates of digestibility for transgenic proteins expressed in crops are required for making a human-health risk assessment by regulatory authorities. The estimation of first-order rate constants for digestion under conditions of low substrate concentration was explored for two protein substrates (azocoll and DQ-ovalbumin). Data conformed to first-order kinetics, and half-lives were relatively insensitive to significant variations in both substrate and pepsin concentration when high purity pepsin preparations were used. Estimation of digestion efficiency using densitometric measurements of relative protein concentration based on SDS-PAGE corroborated digestion estimates based on measurements of dye or fluorescence release from the labeled substrates. The suitability of first-order rate constants for estimating the efficiency of the pepsin digestion of novel proteins is discussed. Results further support a kinetic approach as appropriate for comparing the digestibility of proteins in simulated gastric fluid.

Bone Augmentation in Rabbit Tibia Using Microfixed Cobalt-Chromium Membranes with Whole Blood and Platelet-Rich Plasma.

PubMed

Decco, Oscar A; Beltrán, Víctor; Zuchuat, Jésica I; Cura, Andrea C; Lezcano, María F; Engelke, Wilfried

2015-07-30

Bone augmentation is a subject of intensive investigation in regenerative bone medicine and constitutes a clinical situation in which autogenous bone grafts or synthetic materials are used to aid new bone formation. Based on a non-critical defect, Co-Cr barrier membranes were placed on six adult Fauve de Bourgogne rabbits, divided into two groups: whole blood and PRP. Three densitometric controls were performed during the experiment. The animals were euthanized at 30, 45, 60, and 110 days. The presence of newly formed bone was observed. Samples for histological studies were taken from the augmentation center. External and internal bone tissue augmentation was observed in almost all cases. Significant differences between PRP- and whole blood-stimulated bone augmentation were not observed. At 60 days, bones with PRP presented higher angiogenesis, which may indicate more proliferation and cellular activity. PRP activates the bone regeneration process under optimized conditions by stimulation of osteoblast proliferation after six weeks, when a significant difference in cellular activity was observed. Membranes could stimulate bone augmentation at the site of placement and in the surrounding areas.

Automated characterization of normal and pathologic lung tissue by topological texture analysis of multidetector CT

NASA Astrophysics Data System (ADS)

Boehm, H. F.; Fink, C.; Becker, C.; Reiser, M.

2007-03-01

Reliable and accurate methods for objective quantitative assessment of parenchymal alterations in the lung are necessary for diagnosis, treatment and follow-up of pulmonary diseases. Two major types of alterations are pulmonary emphysema and fibrosis, emphysema being characterized by abnormal enlargement of the air spaces distal to the terminal, nonrespiratory bronchiole, accompanied by destructive changes of the alveolar walls. The main characteristic of fibrosis is coursening of the interstitial fibers and compaction of the pulmonary tissue. With the ability to display anatomy free from superimposing structures and greater visual clarity, Multi-Detector-CT has shown to be more sensitive than the chest radiograph in identifying alterations of lung parenchyma. In automated evaluation of pulmonary CT-scans, quantitative image processing techniques are applied for objective evaluation of the data. A number of methods have been proposed in the past, most of which utilize simple densitometric tissue features based on the mean X-ray attenuation coefficients expressed in terms of Hounsfield Units [HU]. Due to partial volume effects, most of the density-based methodologies tend to fail, namely in cases, where emphysema and fibrosis occur within narrow spatial limits. In this study, we propose a methodology based upon the topological assessment of graylevel distribution in the 3D image data of lung tissue which provides a way of improving quantitative CT evaluation. Results are compared to the more established density-based methods.

Emerging differential roles of the pRb tumor suppressor in trichodysplasia spinulosa-associated polyomavirus and Merkel cell polyomavirus pathogeneses.

PubMed

Wu, Julie H; Simonette, Rebecca A; Nguyen, Harrison P; Doan, Hung Q; Rady, Peter L; Tyring, Stephen K

2016-03-01

Merkel cell carcinoma (MCC) and trichodysplasia spinulosa (TS) are two proliferative cutaneous diseases caused by the Merkel cell polyomavirus (MCPyV) and trichodysplasia spinulosa-associated polyomavirus (TSPyV) respectively. Recently, studies have elucidated a key role of the small tumor (sT) antigen in the proliferative pathogenic mechanisms of MCPyV and likely TSPyV. While both sT antigens have demonstrated a capacity in regulating cellular pathways, it remains unknown whether MCPyV and TSPyV sT antigens contribute similarly or differentially to cell proliferation. The present study aims to explore the proliferative potential of MCPyV and TSPyV sT antigens by investigating their regulatory effects on the retinoblastoma protein (pRb) tumor suppressor. Inducible cell lines expressing MCPyV sT or TSPyV sT were created using a lentiviral packaging system. Cellular proteins were extracted and subjected to SDS-PAGE followed by Western blot detection and densitometric analysis. Expression of TSPyV sT markedly enhanced the phosphorylation of pRb in Western blot experiments. In contrast, expression of MCPyV sT did not alter pRb phosphorylation under the same experimental conditions. Densitometric analysis revealed that TSPyV sT antigen expression nearly doubled the ratio of phosphorylated to total pRb (P<0.001, Student's T-test), while MCPyV sT antigen expression did not cause significant change in pRb phosphorylation status. Given that hyperphosphorylation of pRb is associated with dysregulation of the cell cycle, S-phase induction, and increased cell proliferation, our findings support an important role of TSPyV-mediated pRb deactivation in the development of TS. The observation that the pRb tumor suppressor is inactivated by TSPyV sT but not MCPyV sT provides further insights into the distinct pathobiological mechanisms of MCC and TS. Copyright © 2016 Elsevier B.V. All rights reserved.

A Computer Simulation for Predicting the Time Course of Thermal and Cardiovascular Responses to Various Combinations of Heat Stress, Clothing and Exercise

DTIC Science & Technology

1991-06-01

Anderson and A. Keys, Densitometric analysis of body composition : revision of some quantitative assumptions. Ann. N.Y. Acad. Sci. 110: 113-140, 1963. 6...cylinder at midpoint between adjacent compartments [cm] A, = effective radiating area of the body surface [M’] BF,, n rate of blood flow through...Sutalation for Predicting the Time Cou~rse of ’Thermal and Cardiovrascular Responses to varicus Cmtinations of Heat Stresso Clothing and Excercise 6. AUTHOR

Direct Visualization of Surfaces from Computed Tomography Data,

DTIC Science & Technology

1988-01-01

dominant method - slice-by-slice - makes comprehension of convoluted, small, or faint structures difficult. From a densitometric point of view, the human ...Di t SPeclal 4 N(xI) = Vf(lx) IVf (XI)I* There are many ways to estimate the gradient vector Vf(xi). The selection of an operator depends on the...the data is relatively noise-free or has been pre-smoothed. A graph of a(xi) as a function of f(xi) and IVf (xl)l for three tissue types A, B, and C

Genome size and chromosome number in velvet worms (Onychophora).

PubMed

Jeffery, Nicholas W; Oliveira, Ivo S; Gregory, T Ryan; Rowell, David M; Mayer, Georg

2012-12-01

The Onychophora (velvet worms) represents a small group of invertebrates (~180 valid species), which is commonly united with Tardigrada and Arthropoda in a clade called Panarthropoda. As with the majority of invertebrate taxa, genome size data are very limited for the Onychophora, with only one previously published estimate. Here we use both flow cytometry and Feulgen image analysis densitometry to provide genome size estimates for seven species of velvet worms from both major subgroups, Peripatidae and Peripatopsidae, along with karyotype data for each species. Genome sizes in these species range from roughly 5-19 pg, with densitometric estimates being slightly larger than those obtained by flow cytometry for all species. Chromosome numbers range from 2n = 8 to 2n = 54. No relationship is evident between genome size, chromosome number, or reproductive mode. Various avenues for future genomic research are presented based on these results.

Selective determination of aloin in different matrices by HPTLC densitometry in fluorescence mode.

PubMed

Coran, Silvia A; Bartolucci, Gianluca; Bambagiotti-Alberti, Massimo

2011-01-25

A novel method based on the fluorescence excited solely on aloin by a H₃BO₃ derivatizing procedure, allowed its rapid and selective determination among the co-occurring components in a variety of complex matrices as several Aloes dried extracts and related commercial products. HPTLC LiChrospher silica gel 60 F254S, 20 cm x 10 cm, plates with ethyl formate: CH₃OH:H₂O (100:14.5:10, v/v) as the mobile phase were used. Densitometric determinations were performed in fluorescence mode, exciting wavelength 365 nm, optical filter K540 after derivatization with H₃BO₃. The method was validated giving rise to a dependable and high throughput procedure well suited to routine application. Aloin was quantified in the range of 110-330 ng with RSD of repeatability and intermediate precision not exceeding 2.3% and accuracy inside the acceptance limits. Copyright © 2010 Elsevier B.V. All rights reserved.

[Can the degree of renal artery stenosis be automatically quantified?].

PubMed

Cherrak, I; Jaulent, M C; Azizi, M; Plouin, P F; Degoulet, P; Chatellier, G

2000-08-01

The objective of the reported study is to validate a computer system, QUASAR, dedicated to the quantification of renal artery stenoses. This system estimates automatically the reference diameter and calculates the minimum diameter to compute a degree of stenosis. A hundred and eighty images of atheromatous stenoses between 10% and 80% were collected from two French independent protocols. For the 49 images of the EMMA protocol, the results from QUASAR were compared with the visual estimation of an initial investigator and with the results from a reference method based on a panel of fixe experienced experts. For the 131 images of the ASTARTE protocol, the results from QUASAR were compared with those from a semi-automatic quantification system and with those from a system based on densitometric analysis. The present work validates QUASAR in a population of narrow atheromatous stenoses (> 50%). In the context of the EMMA protocol, QUASAR is not significantly different from the mean of the fixe experts. It is unbiased and more precise than the estimation of a single investigator. In the context of the ASTARTE protocol, there is no significant difference between the three methods for the stenoses higher than 50%, however, globally, QUASAR surestimates significantly (up to 10%) the degree of stenosis.

Different Spectrophotometric and Chromatographic Methods for Determination of Mepivacaine and Its Toxic Impurity.

PubMed

Abdelwahab, Nada S; Fared, Nehal F; Elagawany, Mohamed; Abdelmomen, Esraa H

2017-09-01

Stability-indicating spectrophotometric, TLC-densitometric, and ultra-performance LC (UPLC) methods were developed for the determination of mepivacaine HCl (MEP) in the presence of its toxic impurity, 2,6-dimethylanaline (DMA). Different spectrophotometric methods were developed for the determination of MEP and DMA. In a dual-wavelength method combined with direct spectrophotometric measurement, the absorbance difference between 221.4 and 240 nm was used for MEP measurements, whereas the absorbance at 283 nm was used for measuring DMA in the binary mixture. In the second-derivative method, amplitudes at 272.2 and 232.6 nm were recorded and used for the determination of MEP and DMA, respectively. The developed TLC-densitometric method depended on chromatographic separation using silica gel 60 F254 TLC plates as a stationary phase and methanol-water-acetic acid (9 + 1 + 0.1, v/v/v) as a developing system, with UV scanning at 230 nm. The developed UPLC method depended on separation using a C18 column (250 × 4.6 mm id, 5 μm particle size) as a stationary phase and acetonitrile-water (40 + 60, v/v; pH 4 with phosphoric acid) as a mobile phase at a flow rate of 0.4 mL/min, with UV detection at 215 nm. The chromatographic run time was approximately 1 min. The proposed methods were validated with respect to International Conference on Harmonization guidelines regarding precision, accuracy, ruggedness, robustness, and specificity.

Simultaneous densitometric determination of anthelmintic drug albendazole and its metabolite albendazole sulfoxide by HPTLC in human plasma and pharmaceutical formulations.

PubMed

Pandya, Jui J; Sanyal, Mallika; Shrivastav, Pranav S

2017-09-01

A new, simple, accurate and precise high-performance thin-layer chromatographic method has been developed and validated for simultaneous determination of an anthelmintic drug, albendazole, and its active metabolite albendazole, sulfoxide. Planar chromatographic separation was performed on aluminum-backed layer of silica gel 60G F 254 using a mixture of toluene-acetonitrile-glacial acetic acid (7.0:2.9:0.1, v/v/v) as the mobile phase. For quantitation, the separated spots were scanned densitometrically at 225 nm. The retention factors (R f ) obtained under the established conditions were 0.76 ± 0.01 and 0.50 ± 0.01 and the regression plots were linear (r 2  ≥ 0.9997) in the concentration ranges 50-350 and 100-700 ng/band for albendazole and albendazole sulfoxide, respectively. The method was validated for linearity, specificity, accuracy (recovery) and precision, repeatability, stability and robustness. The limit of detection and limit of quantitation found were 9.84 and 29.81 ng/band for albendazole and 21.60 and 65.45 ng/band for albendazole sulfoxide, respectively. For plasma samples, solid-phase extraction of analytes yielded mean extraction recoveries of 87.59 and 87.13% for albendazole and albendazole sulfoxide, respectively. The method was successfully applied for the analysis of albendazole in pharmaceutical formulations with accuracy ≥99.32%. Copyright © 2017 John Wiley & Sons, Ltd.

Novel Role of Merlin Tumor Suppressor in Autophagy and its Implication in Treating NF2-Associated Tumors

DTIC Science & Technology

2014-04-01

with the idea that Merlin serves as an adaptor linking LC3 to DIC. Moreover, Merlin-KD led to reduce levels of p62 degradation and LC3 -II accumulation...autophagy flux In response to nutrient starvation (Stv), Merlin-knockdown (KD) MEFs show attenuated levels of LC3 -II accumulation or p62 ...degradation, as compared with control MEFs (Cr-KD). Graphs: Densitometric analysis of LC3 -II and p62 on Western blots. Pixel intensities of LC3 -II and p62

Application of spectrophotometric, densitometric, and HPLC techniques as stability indicating methods for determination of Zaleplon in pharmaceutical preparations

NASA Astrophysics Data System (ADS)

Metwally, Fadia H.; Abdelkawy, M.; Abdelwahab, Nada S.

2007-12-01

Spectrophotometric, spectrodensitometric and HPLC are stability indicating methods described for determination of Zaleplon in pure and dosage forms. As Zaleplon is easily degradable, the proposed techniques in this manuscript are adopted for its determination in presence of its alkaline degradation product, namely N-[4-(3-cyano-pyrazolo[1,5a]pyridin-7-yl)-phenyl]- N-ethyl-acetamide. These approaches are successfully applied to quantify Zaleplon using the information included in the absorption spectra of appropriate solutions. The second derivative (D 2) spectrophotometric method, allows determination of Zaleplon without interference of its degradate at 235.2 nm using 0.01N HCl as a solvent with obedience to Beer's law over a concentration range of 1-10 μg ml -1 with mean percentage recovery 100.24 ± 0.86%. The first derivative of the ratio spectra ( 1DD) based on the simultaneous use of ( 1DD) and measurement at 241.8 nm using the same solvent and over the same concentration range as (D 2) spectrophotometric method, with mean percentage recovery 99.9 ± 1.07%. The spectrodensitometric analysis allows the separation and quantitation of Zaleplon from its degradate on silica gel plates using chloroform:acetone:ammonia solution (9:1:0.2 by volume) as a mobile phase. This method depends on quantitave densitometric evaluation of thin layer chromatogram of Zaleplon at 338 nm over a concentration range of 0.2-1 μg band -1, with mean percentage recovery 99.73 ± 1.35. Also a reversed-phase liquid chromatographic method using 5-C8 (22 cm × 4.6 mm i.d. 5 μm particle size) column was described and validated for quantitation of Zaleplon using acetonitrile:deionised water (35:65, v/v) as a mobile phase using Paracetamol as internal standard and a flow rate of 1.5 ml min -1 with UV detection of the effluent at 232 nm at ambient temperature over a concentration range of 2-20 μg ml -1 with mean percentage recovery 100.19 ± 1.15%. The insignificance difference of the proposed methods results with those of the reference one proved their accuracy and precision.

Bone Augmentation in Rabbit Tibia Using Microfixed Cobalt-Chromium Membranes with Whole Blood and Platelet-Rich Plasma

PubMed Central

Decco, Oscar A.; Beltrán, Víctor; Zuchuat, Jésica I.; Cura, Andrea C.; Lezcano, María F.; Engelke, Wilfried

2015-01-01

Background: Bone augmentation is a subject of intensive investigation in regenerative bone medicine and constitutes a clinical situation in which autogenous bone grafts or synthetic materials are used to aid new bone formation. Method: Based on a non-critical defect, Co-Cr barrier membranes were placed on six adult Fauve de Bourgogne rabbits, divided into two groups: whole blood and PRP. Three densitometric controls were performed during the experiment. The animals were euthanized at 30, 45, 60, and 110 days. The presence of newly formed bone was observed. Samples for histological studies were taken from the augmentation center. Results: External and internal bone tissue augmentation was observed in almost all cases. Significant differences between PRP- and whole blood–stimulated bone augmentation were not observed. At 60 days, bones with PRP presented higher angiogenesis, which may indicate more proliferation and cellular activity. Conclusion: PRP activates the bone regeneration process under optimized conditions by stimulation of osteoblast proliferation after six weeks, when a significant difference in cellular activity was observed. Membranes could stimulate bone augmentation at the site of placement and in the surrounding areas. PMID:28793476

Qualitative and quantitative analysis of four species of Curcuma rhizomes using twice development thin layer chromatography.

PubMed

Zhang, J S; Guan, J; Yang, F Q; Liu, H G; Cheng, X J; Li, S P

2008-11-04

The rhizomes of Curcuma phaeocaulis, Curcuma kwangsiensis, Curcuma wenyujin and Curcuma longa are used as Ezhu or Jianghuang in traditional Chinese medicine for a long time. Due to their similar morphological characters, it is difficult to distinguish their origins of raw materials used in clinic. In this study, a simple, rapid and reliable twice development TLC method was developed for qualitative and quantitative analysis of the four species of Curcuma rhizomes. The chromatography was performed on silica gel 60F(254) plate with chloroform-methanol-formic acid (80:4:0.8, v/v/v) and petroleum ether-ethyl acetate (90:10, v/v) as mobile phase for twice development. The TLC markers were colorized with 1% vanillin-H(2)SO(4) solution. The four species of Curcuma were easily discriminated based on their characteristic TLC profiles, and simultaneous quantification of eight compounds, including bisdemethoxycurcumin, demethoxycurcumin, curcumine, curcumenol, curcumol, curdione, furanodienone and curzerene, in Curcuma were also performed densitometrically at lambda(scan)=518nm and lambda(reference)=800 nm. The investigated compounds had good linearity (r(2)>0.9905) within test ranges. Therefore, the developed TLC method can be used for quality control of Curcuma rhizomes.

Application of LANDSAT data to wetland study and land use classification in west Tennessee

NASA Technical Reports Server (NTRS)

Jones, N. L.; Shahrokhi, F.

1977-01-01

The Obion-Forked Deer River Basin in northwest Tennessee is confronted with several acute land use problems which result in excessive erosion, sedimentation, pollution, and hydrologic runoff. LANDSAT data was applied to determine land use of selected watershed areas within the basin, with special emphasis on determining wetland boundaries. Densitometric analysis was performed to allow numerical classification of objects observed in the imagery on the basis of measurements of optical densities. Multispectral analysis of the LANDSAT imagery provided the capability of altering the color of the image presentation in order to enhance desired relationships. Manual mapping and classification techniques were performed in order to indicate a level of accuracy of the LANDSAT data as compared with high and low altitude photography for land use classification.

[CIP and CAP fragments of parathormone and selected parameters of calcium-phosphate balance in patients with chronic kidney disease treated with repeated haemodialysis].

PubMed

Polak-Jonkisz, Dorota; Zwolińska, Danuta; Nahaczewska, Wiesława

2010-01-01

Chronic kidney disease (CKD) leads to bone and mineral complications, which are manifested, among others, by hyperparathyroidism, calcium-phosphate and vitamin D balance disturbances. The results of investigation assessing the usefulness of CAP/CIP ratio, (cyclase activating PTH/cyclase inactive PTH) as a marker of bone turnover and bone disturbances in this group of patients are contradictory. was to estimate the concentration of CAP and CIP of parathormone, connection with selected calcium-phosphate balance parameters and usefulness of CAP/CIP ratio to differentiate bone mineral density in patients with CKD treated with repeated haemodialysis. The study included 31 children aged 5 to 18 years. Group I - 15 haemodialysed children. Group II - 16 healthy children. The patients underwent the following serum measurements: calcium concentration (Ca), inorganic phosphate (P), 1.25-dihydroxyvitamin D, parathormone (intact PTH), and CAP, CIP were evaluated with Scantibodies Laboratory Inc test. In group I the densitometric examination was done using the Lunar DPX-L system, performing the overall bone measurement. In children from group I the average values of iPTH concentration and both CIP and CAP components were significantly elevated (p<0.05) as compared to group II. CAP/CIP ratio in group I was <1; in healthy children >1. Average concentrations of Ca and 1.25(OH)2D in serum of group I were lowered, although without statistical significance in comparison with group II. CAP/CIP ratio does not differentiate the children with bone disturbances. Densitometric examination revealed osteopenic changes in 3 children and osteoporosis in 2 children. There were no statistically significant correlations between the examined parameters. 1. The CIP/CAP ratio does not differentiate the bone mineral density status and it is not associated with biochemical parameters of calcium-phosphate metabolism. 2. This indicates its poor diagnostic utility with reference to mineralization disturbances in children with chronic kidney disease.

Does Guided Bone Regeneration Prevent Unfavorable Bone Shapes in Distraction Gap?

PubMed

Demetoglu, Umut; Alkan, Alper; Kiliç, Erdem; Ozturk, Mustafa; Bilge, Suheyb

2018-03-01

Complications related to distraction osteogenesis can cause degradation of newly regenerated bone. Additionally, an unfavorable shape of the regenerated bone at the distraction gap can reduce the quantity of regenerated bone. The aim of the present study was to report on the prevention of unfavorable shapes of regenerated bone using guided bone regeneration during distraction. Bilateral alveolar distraction was performed in 10 beagle dog mandibles. One side of the mandible formed the experimental group and the other side served as the control group. In the experimental group, guided bone regeneration was performed simultaneously with distraction osteogenesis. In the control group, only alveolar distraction was applied. At the end of a 1-week latent period, all mandibles were distracted 10 mm (1 mm/day). After the distraction period, 3 months were allowed for consolidation. After consolidation, all the dogs were euthanized, and the shape of the regenerated bone was determined to be either favorable or unfavorable. Densitometric evaluation and area measurements were performed using computed tomography scans. Statistical evaluation was performed using the independent t test, with a significance level of P < .05. In the experimental group, no unfavorable bone shape developed in the distraction gap, and the new bone had a surface and volume similar to those of the segments. In contrast, in the control group, 4 mandibles had an unfavorable bone shape in the distraction gap and 4 showed favorable bone healing with no defect. The surface area of the regenerating bone in the experimental group was significantly greater than that in the control group. Also, the surface area differed significantly between the experimental and control groups (P < .05). However, the densitometric values did not differ between the 2 groups (P < .05). Concomitant use of guided bone regeneration with distraction osteogenesis could be an optimal method for generating a favorable bone shape within the distraction gap. Copyright © 2017 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.

Identification of serum protein markers for early diagnosis of pregnancy in buffalo.

PubMed

Buragohain, Lukumoni; Nanda, Trilok; Ghosh, Arnab; Ghosh, Mayukh; Kumar, Rajesh; Kumar, Sunil; Gupta, Sambhu Sharan; Bharali, Arpita; Mohanty, Ashok K; Singh, Inderjeet; Balhara, Ashok Kumar

2017-08-01

Improper or delayed pregnancy diagnosis has significant impact over animal production, particularly in buffaloes which inherently suffer from several reproductive inefficiencies. Thus the present study has undertaken to identify serum protein markers pertaining to early pregnancy diagnosis in buffaloes. Serum samples were collected from 10 pregnant Murrah Buffalo heifers at weekly intervals from days 0-35 post-artificial insemination and from 12 inseminated non-pregnant cyclic buffalo heifers on days 0, 7, 14 and 21. Two-dimensional gel electrophoresis and densitometric analysis revealed the presence of five protein spots showing average density fold change of ≥4 during early pregnancy. Mass spectrometry analysis identified these up-regulated proteins as anti-testosterone antibody light chain, apolipoprotein A-II precursor, serum amyloid A, cytokeratin type II, component IV isoform 1, which are have established roles in embryogenesis, but over-expression of the fifth identified protein immunoglobulin lambda light chain in pregnancy has been elucidated as a novel finding in the current study. Further, with bioinformatics analysis, potential antigenic B-cell epitopes were predicted for all these five proteins. An antibody cocktail-based approach involving antibodies against all these five up-regulated entire proteins or their epitopes could be developed for early detection of pregnancy in buffaloes. © 2016 Japanese Society of Animal Science. © 2016 Japanese Society of Animal Science.

Stability indicating spectrophotometric and spectrodensitometric methods for the determination of diatrizoate sodium in presence of its degradation product

NASA Astrophysics Data System (ADS)

Abd El-Rahman, Mohamed K.; Riad, Safaa M.; Abdel Gawad, Sherif A.; Fawaz, Esraa M.; Shehata, Mostafa A.

2015-02-01

Three sensitive, selective, and precise stability indicating methods for the determination of the X-ray contrast agent, diatrizoate sodium (DTA), in the presence of its acidic degradation product (highly cytotoxic 3,5 diamino metabolite) and in pharmaceutical formulation were developed and validated. The first method is a first derivative (D1) spectrophotometric one, which allows the determination of DTA in the presence of its degradate at 231.2 nm (corresponding to zero crossing of the degradate) over a concentration range of 2-24 μg/mL with mean percentage recovery 99.95 ± 0.97%. The second method is the first derivative of the ratio spectra (DD1) by measuring the peak amplitude at 227 nm over the same concentration range as D1 spectrophotometric method, with mean percentage recovery 99.99 ± 1.15%. The third method is a TLC-densitometric one, where DTA was separated from its degradate on silica gel plates using chloroform:methanol:ammonium hydroxide (20:10:2 by volume) as a developing system. This method depends on quantitative densitometric evaluation of thin layer chromatogram of DTA at 238 nm over a concentration range of 4-20 μg/spot, with mean percentage recovery 99.88 ± 0.89%. The selectivity of the proposed methods was tested using laboratory-prepared mixtures. The proposed methods have been successfully applied to the analysis of DTA in pharmaceutical dosage forms without interference from other dosage form additives. The results were statistically compared with the official US pharmacopeial method. No significant difference for either accuracy or precision was observed.

DNA fragmentation and nuclear phenotype in tendons exposed to low-intensity infrared laser

NASA Astrophysics Data System (ADS)

de Paoli, Flavia; Ramos Cerqueira, Larissa; Martins Ramos, Mayara; Campos, Vera M.; Ferreira-Machado, Samara C.; Geller, Mauro; de Souza da Fonseca, Adenilson

2015-03-01

Clinical protocols are recommended in device guidelines outlined for treating many diseases on empirical basis. However, effects of low-intensity infrared lasers at fluences used in clinical protocols on DNA are controversial. Excitation of endogenous chromophores in tissues and free radicals generation could be described as a consequence of laser used. DNA lesions induced by free radicals cause changes in DNA structure, chromatin organization, ploidy degrees and cell death. In this work, we investigated whether low-intensity infrared laser therapy could alter the fibroblasts nuclei characteristics and induce DNA fragmentation. Tendons of Wistar rats were exposed to low-intensity infrared laser (830 nm), at different fluences (1, 5 and 10 J/cm2), in continuous wave (power output of 10mW, power density of 79.6 mW/cm2). Different frequencies were analyzed for the higher fluence (10 J/cm2), at pulsed emission mode (2.5, 250 and 2500 Hz), with the laser source at surface of skin. Geometric, densitometric and textural parameters obtained for Feulgen-stained nuclei by image analysis were used to define nuclear phenotypes. Significant differences were observed on the nuclear phenotype of tendons after exposure to laser, as well as, high cell death percentages was observed for all fluences and frequencies analyzed here, exception 1 J/cm2 fluence. Our results indicate that low-intensity infrared laser can alter geometric, densitometric and textural parameters in tendon fibroblasts nuclei. Laser can also induce DNA fragmentation, chromatin lost and consequently cell death, using fluences, frequencies and emission modes took out from clinical protocols.

Densitometric and biochemical values of broiler tibias at different ages.

PubMed

Barreiro, F R; Sagula, A L; Junqueira, O M; Pereira, G T; Baraldi-Artoni, S M

2009-12-01

The objective of this experiment was to determine the normal values of bone radiographic density (BRD) by using the optical densitometry in radiographic images and the biochemical values represented by serum calcium, ash percentage, and minerals (calcium, phosphorus, and magnesium) from tibia ash of Cobb broilers at 8, 22, and 43 d of age. A total of 14 broilers were used for densitometric analysis, and 15 were used for biochemical dosages. The BRD values increased (P < 0.05) with age and in all tibia regions (proximal epiphysis, diaphysis, and distal epiphysis), concluding that growth was a determinative factor for bone performance, demanding a higher BRD during broiler development. Tibia proximal epiphysis presented higher BRD values in relation to the other bone regions (P < 0.05), as a result of a possible biomechanical adaptation to ligaments and tension of the muscle tendons at this region, allowing the support of the muscle mass increase. The serum calcium values were kept constant, as a result of the appropriate nutritional levels of the diet that supported the animal homeostasis. The bone ash and mineral percentage increased (P < 0.05) at 22 d of age, due to the higher mineral requirement in this age. The correlation between bone densitometry and the invasive techniques showed that the bone densitometry can substitute the determination of mineral percentage in the ash. This experiment presented normal values of the noninvasive and invasive methods more used in aviculture, allowing us to compare, subsequently, pathological and physiological values or results of broilers fed with different diets.

Marshes and turbid waters in the French Atlantic littoral

NASA Technical Reports Server (NTRS)

Verger, F. (Principal Investigator)

1973-01-01

The author has identified the following significant results. The multispectral information provided by ERTS-1 is very rich for the coastal regions but the cloud cover, even when only partial, often cuts up the data, and lessens its practical value. The research by densitometric methods has thus far been the most fruitful. It consists of measuring densities along preferential axes. This method has enabled the investigators to perfect a system of computer cartography for the best image. Comparative study of microdensitometric transects in four MSS bands for carefully dilimited profiles on the ground, makes it possible to isolate and recognize various types of countryside: (1) countryside of coastal plains; (2) countryside of coastal sand dunes and beaches; and (3) forms and processes of offshore domains. This study shows the usefulness of the ERTS program in establishing a rapid cartography of the physiographic units of the coastal plains in the interest of a rational program of exploitation and development.

Prediction of Hip Failure Load: In Vitro Study of 80 Femurs Using Three Imaging Methods and Finite Element Models-The European Fracture Study (EFFECT).

PubMed

Pottecher, Pierre; Engelke, Klaus; Duchemin, Laure; Museyko, Oleg; Moser, Thomas; Mitton, David; Vicaut, Eric; Adams, Judith; Skalli, Wafa; Laredo, Jean Denis; Bousson, Valérie

2016-09-01

Purpose To evaluate the performance of three imaging methods (radiography, dual-energy x-ray absorptiometry [DXA], and quantitative computed tomography [CT]) and that of a numerical analysis with finite element modeling (FEM) in the prediction of failure load of the proximal femur and to identify the best densitometric or geometric predictors of hip failure load. Materials and Methods Institutional review board approval was obtained. A total of 40 pairs of excised cadaver femurs (mean patient age at time of death, 82 years ± 12 [standard deviation]) were examined with (a) radiography to measure geometric parameters (lengths, angles, and cortical thicknesses), (b) DXA (reference standard) to determine areal bone mineral densities (BMDs), and (c) quantitative CT with dedicated three-dimensional analysis software to determine volumetric BMDs and geometric parameters (neck axis length, cortical thicknesses, volumes, and moments of inertia), and (d) quantitative CT-based FEM to calculate a numerical value of failure load. The 80 femurs were fractured via mechanical testing, with random assignment of one femur from each pair to the single-limb stance configuration (hereafter, stance configuration) and assignment of the paired femur to the sideways fall configuration (hereafter, side configuration). Descriptive statistics, univariate correlations, and stepwise regression models were obtained for each imaging method and for FEM to enable us to predict failure load in both configurations. Results Statistics reported are for stance and side configurations, respectively. For radiography, the strongest correlation with mechanical failure load was obtained by using a geometric parameter combined with a cortical thickness (r(2) = 0.66, P < .001; r(2) = 0.65, P < .001). For DXA, the strongest correlation with mechanical failure load was obtained by using total BMD (r(2) = 0.73, P < .001) and trochanteric BMD (r(2) = 0.80, P < .001). For quantitative CT, in both configurations, the best model combined volumetric BMD and a moment of inertia (r(2) = 0.78, P < .001; r(2) = 0.85, P < .001). FEM explained 87% (P < .001) and 83% (P < .001) of bone strength, respectively. By combining (a) radiography and DXA and (b) quantitative CT and DXA, correlations with mechanical failure load increased to 0.82 (P < .001) and 0.84 (P < .001), respectively, for radiography and DXA and to 0.80 (P < .001) and 0.86 (P < .001) , respectively, for quantitative CT and DXA. Conclusion Quantitative CT-based FEM was the best method with which to predict the experimental failure load; however, combining quantitative CT and DXA yielded a performance as good as that attained with FEM. The quantitative CT DXA combination may be easier to use in fracture prediction, provided standardized software is developed. These findings also highlight the major influence on femoral failure load, particularly in the trochanteric region, of a densitometric parameter combined with a geometric parameter. (©) RSNA, 2016 Online supplemental material is available for this article.

May–June Maximum Temperature Reconstruction from Mean Earlywood Density in North Central China and Its Linkages to the Summer Monsoon Activities

PubMed Central

Chen, Feng; Yuan, Yujiang

2014-01-01

Cores of Pinus tabulaformis from Tianshui were subjected to densitometric analysis to obtain mean earlywood density data. Climate response analysis indicates that May–June maximum temperature is the main factor limiting the mean earlywood density (EWD) of Chinese pine trees in the Shimen Mountains. Based on the EWD chronology, we have reconstructed May–June maximum temperature 1666 to 2008 for Tianshui, north central China. The reconstruction explains 40.1% of the actual temperature variance during the common period 1953–2008. The temperature reconstruction is representative of temperature conditions over a large area to the southeast and northwest of the sampling site. Preliminary analysis of links between large-scale climatic variation and the temperature reconstruction shows that there is a relationship between extremes in spring temperature and anomalous atmospheric circulation in the region. It is thus revealed that the mean earlywood density chronology of Pinus tabulaformis has enough potential to reconstruct the temperature variability further into the past. PMID:25207554

Systematic Development and Validation of a Thin-Layer Densitometric Bioanalytical Method for Estimation of Mangiferin Employing Analytical Quality by Design (AQbD) Approach

PubMed Central

Khurana, Rajneet Kaur; Rao, Satish; Beg, Sarwar; Katare, O.P.; Singh, Bhupinder

2016-01-01

The present work aims at the systematic development of a simple, rapid and highly sensitive densitometry-based thin-layer chromatographic method for the quantification of mangiferin in bioanalytical samples. Initially, the quality target method profile was defined and critical analytical attributes (CAAs) earmarked, namely, retardation factor (Rf), peak height, capacity factor, theoretical plates and separation number. Face-centered cubic design was selected for optimization of volume loaded and plate dimensions as the critical method parameters selected from screening studies employing D-optimal and Plackett–Burman design studies, followed by evaluating their effect on the CAAs. The mobile phase containing a mixture of ethyl acetate : acetic acid : formic acid : water in a 7 : 1 : 1 : 1 (v/v/v/v) ratio was finally selected as the optimized solvent for apt chromatographic separation of mangiferin at 262 nm with Rf 0.68 ± 0.02 and all other parameters within the acceptance limits. Method validation studies revealed high linearity in the concentration range of 50–800 ng/band for mangiferin. The developed method showed high accuracy, precision, ruggedness, robustness, specificity, sensitivity, selectivity and recovery. In a nutshell, the bioanalytical method for analysis of mangiferin in plasma revealed the presence of well-resolved peaks and high recovery of mangiferin. PMID:26912808

Extraction and quantification of gymnemic acids through gymnemagenin from callus cultures of Gymnema sylvestre.

PubMed

Kanetkar, P V; Singhal, R S; Laddha, K S; Kamat, M Y

2006-01-01

The phyto-constituents of Gymnema sylvestre are used in the treatment of diabetes and obesity. The present work reports on the extraction of gymnemic acid through gymnemagenin from callus cultures of G. sylvestre. Components were separated on pre-coated silica gel 60 GF254 plates with chloroform:methanol (8:2) and scanned using a densitometric scanner at 205 nm in the near-UV region. Linearity of determination of gymnemagenin was observed in the range 2-10 microg. The average percentage recovery of gymnemagenin from leaf callus extracts was 98.9+/-0.3.

Dynamics of distribution and density of phreatophytes and other arid land plant communities

NASA Technical Reports Server (NTRS)

Turner, R. M. (Principal Investigator)

1973-01-01

The author has identified the following significant results. Ground truth measurements of plant coverage on six satellite overflight dates reveal unique trends in coverage for the five desert or semi-desert communities selected. Densitometry and multispectral additive color viewing were used in a preliminary analysis of imagery using the electronic satellite image analyzer console at Stanford Research Institute. The densitometric analysis shows promise for mapping boundaries between plant communities. Color additive viewing of a chronologic sequence of the same scene shown in rapid order will provide a method for mapping phreatophyte communities.

Geographic applications of ERTS-1 data to landscape change

NASA Technical Reports Server (NTRS)

Rehder, J. B.

1973-01-01

The analysis of landscape change requires large area coverage on a periodic basis in order to analyze aggregate changes over an extended period of time. To date, only the ERTS program can provide this capability. Three avenues of experimentation and analysis are being used in the investigation: (1) a multi-scale sampling procedure utilizing aircraft imagery for ground truth and control; (2) a densitometric and computer analytical experiment for the analysis of gray tone signatures, comparisons and ultimately for landscape change detection and monitoring; and (3) an ERTS image enhancement procedure for the detection and analysis of photomorphic regions.

Linear Calibration of Radiographic Mineral Density Using Video-Digitizing Methods

NASA Technical Reports Server (NTRS)

Martin, R. Bruce; Papamichos, Thomas; Dannucci, Greg A.

1990-01-01

Radiographic images can provide quantitative as well as qualitative information if they are subjected to densitometric analysis. Using modem video-digitizing techniques, such densitometry can be readily accomplished using relatively inexpensive computer systems. However, such analyses are made more difficult by the fact that the density values read from the radiograph have a complex, nonlinear relationship to bone mineral content. This article derives the relationship between these variables from the nature of the intermediate physical processes, and presents a simple mathematical method for obtaining a linear calibration function using a step wedge or other standard.

Linear Calibration of Radiographic Mineral Density Using Video-Digitizing Methods

NASA Technical Reports Server (NTRS)

Martin, R. Bruce; Papamichos, Thomas; Dannucci, Greg A.

1990-01-01

Radiographic images can provide quantitative as well as qualitative information if they are subjected to densitometric analysis. Using modern video-digitizing techniques, such densitometry can be readily accomplished using relatively inexpensive computer systems. However, such analyses are made more difficult by the fact that the density values read from the radiograph have a complex, nonlinear relationship to bone mineral content. This article derives the relationship between these variables from the nature of the intermediate physical processes, and presents a simple mathematical method for obtaining a linear calibration function using a step wedge or other standard.

Standardisation of Gymnema sylvestre r. Br. with reference to gymnemagenin by high-performance thin-layer chromatography.

PubMed

Puratchimani, V; Jha, S

2004-01-01

A simple and reproducible HPTLC method for the determination of gymnemagenin (1) in Gymnema sylvestre has been developed. Components were separated on pre-coated silica gel 60 F254 plates with chloroform:methanol (9:1) and scanned using a densitometric scanner in the UV reflectance mode at 290 nm. Linearity of determination of 1 was observed in the range 4-10 microg. The average percentage recovery of 1 from an extract was 99.09 +/- 0.29, and the content of 1 in leaves of the title plant was 1.61% (dry weight).

Identification and Structure Elucidation of Forced Degradation Products of the Novel Propionic acid Derivative Loxoprofen: Development of Stability-Indicating Chromatographic Methods Validated as per ICH Guidelines.

PubMed

Eissa, Maya S; Abd El-Sattar, Osama I

2017-04-01

Loxoprofen sodium (LOX) is a recently developed novel propionic acid derivative. Owing to its instability under both hydrolytic and oxidative conditions, the development of simple, rapid and sensitive methods for its determination in the presence of its possible forced degradation products becomes essential. Two simple chromatographic methods, high-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC), were developed associated with ultraviolet (UV) detection. In HPTLC-densitometric method, the separation of LOX from its degradation products was achieved using silica gel F254 plates and toluene:acetone:acetic acid (1.8:1.0:0.1, v/v/v) as the developing system followed by densitometric scanning at 220 nm. In the HPLC-UV method, the separation was performed using isocratic elution system with acetonitrile: 0.15% triethylamine (pH 2.2) (50:50, v/v) on C18 analytical column. The flow rate was optimized at 1.0 mL·min-1 and UV detection was achieved at 220 nm. Validation was performed in accordance with the International Conference on Harmonization guidelines and the method was perfectly applied for determination of LOX in its pharmaceutical preparation. The results obtained were statistically compared to those obtained after application of the official HPLC method, where no significant difference was found incompliance with precision and accuracy. Identification and characterization of the possible hydrolytic degradation product under alkaline conditions and that produced during oxidative degradation using hydrogen peroxide were structurally elucidated using infrared and mass spectrometry analyses. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Comparison of autogenous cancellous bone grafting and extracorporeal shock wave therapy on osteotomy healing in the tibial tuberosity advancement procedure in dogs. Radiographic densitometric evaluation.

PubMed

Barnes, K; Lanz, O; Werre, S; Clapp, K; Gilley, R

2015-01-01

To compare optical values in the osteotomy gap created after a tibial tuberosity advancement (TTA) treated with autogenous cancellous bone graft, extracorporeal shock wave therapy, a combination of autogenous cancellous bone graft and extracorporeal shock wave therapy, and absence of both autogenous cancellous bone graft and extracorporeal shock wave therapy using densitometry. Dogs that were presented for surgical repair of a cranial cruciate ligament rupture were randomly assigned to one of four groups: TTA with autogenous cancellous bone graft (TTA-G), TTA with autogenous cancellous bone graft and extracorporeal shock wave therapy (TTA-GS), TTA with extracorporeal shock wave therapy (TTA-S), and TTA with no additional therapy (TTA-O). Mediolateral radiographs at zero, four and eight weeks after surgery were evaluated to compare healing of the osteotomy gap via densitometry. An analysis of variance was used to compare the densitometric values between groups. At four weeks after surgery, a significant difference in osteotomy gap density was noted between TTA-GS (8.4 millimetres of aluminium equivalent [mmAleq]) and TTA-S (6.1 mmAleq), and between TTA-GS (8.4 mmAleq) and TTA-O (6.4 mmAleq). There were no significant differences noted between any groups at the eight week re-evaluation. There were no significant differences in the osteotomy gap density at eight weeks after surgery regardless of the treatment modality used. The combination of autogenous cancellous bone graft and extracorporeal shock wave therapy may lead to increased radiographic density of the osteotomy gap in the first four weeks after surgery. Densitometry using an aluminium step wedge is a feasible method for comparison of bone density after TTA in dogs.

Automated brain computed tomographic densitometry of early ischemic changes in acute stroke

PubMed Central

Stoel, Berend C.; Marquering, Henk A.; Staring, Marius; Beenen, Ludo F.; Slump, Cornelis H.; Roos, Yvo B.; Majoie, Charles B.

2015-01-01

Abstract. The Alberta Stroke Program Early CT score (ASPECTS) scoring method is frequently used for quantifying early ischemic changes (EICs) in patients with acute ischemic stroke in clinical studies. Varying interobserver agreement has been reported, however, with limited agreement. Therefore, our goal was to develop and evaluate an automated brain densitometric method. It divides CT scans of the brain into ASPECTS regions using atlas-based segmentation. EICs are quantified by comparing the brain density between contralateral sides. This method was optimized and validated using CT data from 10 and 63 patients, respectively. The automated method was validated against manual ASPECTS, stroke severity at baseline and clinical outcome after 7 to 10 days (NIH Stroke Scale, NIHSS) and 3 months (modified Rankin Scale). Manual and automated ASPECTS showed similar and statistically significant correlations with baseline NIHSS (R=−0.399 and −0.277, respectively) and with follow-up mRS (R=−0.256 and −0.272), except for the follow-up NIHSS. Agreement between automated and consensus ASPECTS reading was similar to the interobserver agreement of manual ASPECTS (differences <1 point in 73% of cases). The automated ASPECTS method could, therefore, be used as a supplementary tool to assist manual scoring. PMID:26158082

[The relationship between the parameters of mineral density of bone tissue and somatotype in women residing in the Republic of Karelia].

PubMed

Pashkova, I G; Gaivoronskiy, I V; Aleksina, L A; Kornev, M A

2014-01-01

Comprehensive anthropometric and densitometric study using the dual x-ray absorptiometry was conducted to determine the relationship between the mineral density of bone tissue and somatotype in 360 women aged 20 to 87 years, permanently residing in the Republic of Karelia. Significant direct correlation was detected between the somatotype and the amount of mineral substances in the vertebrae, bone mineral density and the area of the lumbar vertebrae. Bone mineral density level of the lumbar vertebrae was higher in women with europlastic and athletic somatotypes, which were characterized by high values of body mass and length, body muscle and fat mass. Low values of bone mineral density of vertebrae were identified in women belonging to subathletic, mesoplastic and stenoplastic somatotypes. The risk of developing osteopenia and osteoporosis is increased in women with low body muscle mass.

Complex tibial fracture outcomes following treatment with low-intensity pulsed ultrasound.

PubMed

Leung, Kwok-Sui; Lee, Wing-Sze; Tsui, Hon-For; Liu, Paul Po-Lung; Cheung, Wing-Hoi

2004-03-01

A clinical study was conducted to investigate the effect of low-intensity pulsed ultrasound (US) stimulation (LIPUS) on the healing of complex tibial fractures. Thirty complex tibial fractures were randomly assigned to the treatment with LIPUS (n = 16) or by a dummy machine (sham-exposed: n = 14). The fractures were immobilized by either internal or external fixations according to the clinical indications. LIPUS was given 20 min/day for 90 days. Fracture healing was monitored by clinical, radiological, densitometric and biochemical assessments. The LIPUS-treated group showed statistically significantly better healing, as demonstrated by all assessments. Complications were minimal in the LIPUS group. There were two cases of delayed union, with one in each group. There were two cases of infection in the control group. The delayed-union cases were subsequently treated by LIPUS and the infection cases were treated with standard protocol. Fracture healing in these patients was again treated by LIPUS.

Screening and determination of sibutramine in adulterated herbal slimming supplements by HPTLC-UV densitometry.

PubMed

Mathon, Caroline; Ankli, Anita; Reich, Eike; Bieri, Stefan; Christen, Philippe

2014-01-01

The adulteration of herbal supplements is of growing importance, especially when they contain undeclared compounds like sibutramine that are unsafe drugs. Sibutramine was withdrawn from US and European markets in 2010. In this study, an HPTLC-UV densitometric method was developed for the quantification of sibutramine in herbal diet foods. Sample extracts were directly applied onto HPTLC silica gel plates and separated with a mobile phase made of a toluene-methanol mixture. Sibutramine was quantified at 225 nm and its unequivocal identification was confirmed by MS using a TLC-MS interface. During two surveys, 52 weight loss supplements obtained via the Internet were screened. Half of those were adulterated with sibutramine at amounts reaching up to 35 mg per capsule. The results of this validated HPTLC method were compared with those obtained by HPLC-UV and HPLC-MS/MS. The results were not significantly different with the three methods.

Management of natural resources through automatic cartographic inventory. [France

NASA Technical Reports Server (NTRS)

Rey, P.; Gourinard, Y.; Cambou, F. (Principal Investigator)

1974-01-01

The author has identified the following significant results. (1) Accurate recognition of previously known ground features from ERTS-1 imagery has been confirmed and a probable detection range for the major signatures can be given. (2) Unidentified elements, however, must be decoded by means of the equal densitometric value zone method. (3) Determination of these zonings involves an analogical treatment of images using the color equidensity methods (pseudo-color), color composites and especially temporal color composite (repetitive superposition). (4) After this analogical preparation, the digital equidensities can be processed by computer in the four MSS bands, according to a series of transfer operations from imagery and automatic cartography.

PERFILS: a program for the quantitative treatment of footprinting data.

PubMed

Salas, X; Portugal, J

1993-10-01

PERFILS, a computer program written in Borland TurboPascal, performs quantitative analysis of footprinting experiments using any IBM PC or compatible microcomputer. The program uses the height of the bands obtained from densitometric scanning of footprinting autoradiographs to calculate a differential cleavage plot. Such a plot displays, on a logarithmic scale, the difference of susceptibility of a DNA fragment to DNase I, or any other cleaving agent, in the presence of any ligand versus the sequence. PERFILS calculates the fractional cleavage values for control and ligand, giving a table of values for each internucleotidic bond and rendering the differential cleavage plot in only a few seconds.

Systematic Development and Validation of a Thin-Layer Densitometric Bioanalytical Method for Estimation of Mangiferin Employing Analytical Quality by Design (AQbD) Approach.

PubMed

Khurana, Rajneet Kaur; Rao, Satish; Beg, Sarwar; Katare, O P; Singh, Bhupinder

2016-01-01

The present work aims at the systematic development of a simple, rapid and highly sensitive densitometry-based thin-layer chromatographic method for the quantification of mangiferin in bioanalytical samples. Initially, the quality target method profile was defined and critical analytical attributes (CAAs) earmarked, namely, retardation factor (Rf), peak height, capacity factor, theoretical plates and separation number. Face-centered cubic design was selected for optimization of volume loaded and plate dimensions as the critical method parameters selected from screening studies employing D-optimal and Plackett-Burman design studies, followed by evaluating their effect on the CAAs. The mobile phase containing a mixture of ethyl acetate : acetic acid : formic acid : water in a 7 : 1 : 1 : 1 (v/v/v/v) ratio was finally selected as the optimized solvent for apt chromatographic separation of mangiferin at 262 nm withRf 0.68 ± 0.02 and all other parameters within the acceptance limits. Method validation studies revealed high linearity in the concentration range of 50-800 ng/band for mangiferin. The developed method showed high accuracy, precision, ruggedness, robustness, specificity, sensitivity, selectivity and recovery. In a nutshell, the bioanalytical method for analysis of mangiferin in plasma revealed the presence of well-resolved peaks and high recovery of mangiferin. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

15N-metabolic labeling for comparative plasma membrane proteomics in Arabidopsis cells.

PubMed

Lanquar, Viviane; Kuhn, Lauriane; Lelièvre, Françoise; Khafif, Mehdi; Espagne, Christelle; Bruley, Christophe; Barbier-Brygoo, Hélène; Garin, Jérôme; Thomine, Sébastien

2007-03-01

An important goal for proteomic studies is the global comparison of proteomes from different genotypes, tissues, or physiological conditions. This has so far been mostly achieved by densitometric comparison of spot intensities after protein separation by 2-DE. However, the physicochemical properties of membrane proteins preclude the use of 2-DE. Here, we describe the use of in vivo labeling by the stable isotope 15N as an alternative approach for comparative membrane proteomic studies in plant cells. We confirm that 15N-metabolic labeling of proteins is possible and efficient in Arabidopsis suspension cells. Quantification of 14N versus 15N MS signals reflects the relative abundance of 14N and 15N proteins in the sample analyzed. We describe the use of 15N-metabolic labeling to perform a partial comparative analysis of Arabidopsis cells following cadmium exposure. By focusing our attention on plasma membrane proteins, we were able to confidently identify proteins showing up to 5-fold regulation compared to unexposed cells. This study provides a proof of principle that 15N-metabolic labeling is a useful technique for comparative membrane proteome studies.

Scanning fluorescent microscopy is an alternative for quantitative fluorescent cell analysis.

PubMed

Varga, Viktor Sebestyén; Bocsi, József; Sipos, Ferenc; Csendes, Gábor; Tulassay, Zsolt; Molnár, Béla

2004-07-01

Fluorescent measurements on cells are performed today with FCM and laser scanning cytometry. The scientific community dealing with quantitative cell analysis would benefit from the development of a new digital multichannel and virtual microscopy based scanning fluorescent microscopy technology and from its evaluation on routine standardized fluorescent beads and clinical specimens. We applied a commercial motorized fluorescent microscope system. The scanning was done at 20 x (0.5 NA) magnification, on three channels (Rhodamine, FITC, Hoechst). The SFM (scanning fluorescent microscopy) software included the following features: scanning area, exposure time, and channel definition, autofocused scanning, densitometric and morphometric cellular feature determination, gating on scatterplots and frequency histograms, and preparation of galleries of the gated cells. For the calibration and standardization Immuno-Brite beads were used. With application of shading compensation, the CV of fluorescence of the beads decreased from 24.3% to 3.9%. Standard JPEG image compression until 1:150 resulted in no significant change. The change of focus influenced the CV significantly only after +/-5 microm error. SFM is a valuable method for the evaluation of fluorescently labeled cells. Copyright 2004 Wiley-Liss, Inc.

The Design of a Quantitative Western Blot Experiment

PubMed Central

Taylor, Sean C.; Posch, Anton

2014-01-01

Western blotting is a technique that has been in practice for more than three decades that began as a means of detecting a protein target in a complex sample. Although there have been significant advances in both the imaging and reagent technologies to improve sensitivity, dynamic range of detection, and the applicability of multiplexed target detection, the basic technique has remained essentially unchanged. In the past, western blotting was used simply to detect a specific target protein in a complex mixture, but now journal editors and reviewers are requesting the quantitative interpretation of western blot data in terms of fold changes in protein expression between samples. The calculations are based on the differential densitometry of the associated chemiluminescent and/or fluorescent signals from the blots and this now requires a fundamental shift in the experimental methodology, acquisition, and interpretation of the data. We have recently published an updated approach to produce quantitative densitometric data from western blots (Taylor et al., 2013) and here we summarize the complete western blot workflow with a focus on sample preparation and data analysis for quantitative western blotting. PMID:24738055

Simultaneous Feulgen densitometry and autoradiographic grain counting with the Quantimet 720D image-analysis system. III. Improvements in Feulgen densitometry.

PubMed

Sklarew, R J

1983-10-01

A method has been developed for densitometric estimation of the Feulgen-stained DNA content of 3H-labeled nuclei in autoradiographs in conjunction with automated grain counting using a Quantimet Imaging System. Refinements in the methodology are reported which include 1) the incorporation of an Image-Editor Module into the Quantimet module configuration; 2) the optimization of incident illumination based upon evaluation of various light sources; 3) changes in the optical configuration which reduce glare and minimize the level of monitor shading correction; 4) the optimization of scanner sensitivity; and 5) the evaluation of cell-flattening and staining with respect to densitometry resolution and sensitivity. These refinements resulted in a CV of less than 6.4% in the G-1 and G-2 DNA peaks of rat kidney cells in autoradiographs compared to the previous CV of 10.5%, and a G-2 to G-1 ratio of 2.025. For a fixed field position the CV was 5.1% and the replication error less than 1.0%.

Landsat Image Map Production Methods at the U. S. Geological Survey

USGS Publications Warehouse

Kidwell, R.D.; Binnie, D.R.; Martin, S.

1987-01-01

To maintain consistently high quality in satellite image map production, the U. S. Geological Survey (USGS) has developed standard procedures for the photographic and digital production of Landsat image mosaics, and for lithographic printing of multispectral imagery. This paper gives a brief review of the photographic, digital, and lithographic procedures currently in use for producing image maps from Landsat data. It is shown that consistency in the printing of image maps is achieved by standardizing the materials and procedures that affect the image detail and color balance of the final product. Densitometric standards are established by printing control targets using the pressplates, inks, pre-press proofs, and paper to be used for printing.

A comparison between high-altitude horizon photography and direct sampling of El Chichon cloud particles

NASA Technical Reports Server (NTRS)

Coletti, A.; Hofmann, D. J.; Rosen, J. M.

1986-01-01

Perturbations to the visible radiation by the El Chichon aerosol layers in the stratosphere observed on May 18, 1982 in Laredo, Texas using in situ, time-lapsed photography are analyzed. The densitometric data are compared with optical counter data. Good correlation is detected for the scattered light intensities of the sky estimated with the two techniques. It is observed that the optical thickness of the stratosphere from 18.8 km to the top of the atmosphere = 0.18 and the residual optical thickness at 27 km = 0.0007. The relationship between the isodensity contours and the height of the observations, cloud cover, specific vertical aerosol distribution, and earth curvature is examined.

Applications of ERTS-1 data to landscape change in eastern Tennessee

NASA Technical Reports Server (NTRS)

Rehder, J. B. (Principal Investigator)

1973-01-01

The author has identified the following significant results. The analysis of landscape change in eastern Tennessee from ERTS-1 data is being derived from three avenues of experimentation and analysis: (1) a multi-stage sampling procedure utilizing ground and aircraft imagery for ground truth and control; (2) a densitometric and computer analytical experiment for the analysis of gray tone signatures and comparisons for landscape change detection and monitoring; and (3) an ERTS image enhancement procedure for the detection and analysis of photomorphic regions. Significant results include: maps of strip mining changes and forest inventory, watershed identification and delimitation, and agricultural regions derived from spring plowing patterns appearing on the ERTS-1 imagery.

Cayler cardiofacial syndrome and del 22q11: Part of the CATCH22 phenotype

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

Giannotti, A.; Digilio, M.C.; Marino, B.

1994-11-15

The authors report evidence supporting the hypothesis that del(22)(q11) can be a pathogenetic mechanism for the association between hypoplasia of the depressor anguli oris muscle (DAOM) and conotruncal cardiac malformations. A series of over 180 patients was investigated with deletions of 22q11 with conotruncal defects. About 2/3 of these patients had isolated, nonfamilial cardiac defects. Hemizygosity was searched using the HD7k probe and densitometric analysis. In the patients with molecular evidence of del(22)(q11), hemizygosity was confirmed also using fluorescence in situ hybridization (FISH) with SC11.1 probe. No deletion was found in the parents of hemizygous patients. 16 refs.

Densitometric evaluation of three intra-oral radiographic films.

PubMed

Seeliger, J E; Prinsloo, J J

1989-05-01

The radiographic, or diagnostic, quality of the processed radiograph depends upon a number of factors, one of the most important being the characteristics of the film. The purpose of this study was to determine which of three intra-oral radiographic films, obtainable in this country, would give the best results in terms of density range, speed, contrast and base plus fog values. Agfa Dentus, Flow X-Ray and Kodak (all speed group D films) were exposed, using a calibrated G.E. 1000 x-ray generator at 65 kVp, 10 mA and 50 impulses (1 second) exposure time. The target-film-distance was 40 cm, the total filtration 2.0 mm Aluminium and the half-value layer 2.7 mm Aluminium equivalent. An aluminium step-wedge with 8 steps, in steps of 1.5 mm, and a natural premolar tooth, with a carious lesion, embedded in acrylic, were used as phantoms. An 8 mm-thick layer of base-plate wax and a 3 mm-thick lead plate were used to simulate tissue-scatter and prevent back-scatter, respectively. To determine the base plug fog value, an unexposed film from the same batch was processed simultaneously with each of the three films evaluated. All processing was done in a Dürr AC 245 L processor with automatic replenishment and a 6-minute cycle. The processing chemicals, viz., Kolchem High Stability X-ray developer and fixer, were mixed and used in strict accordance with the manufacturer's recommendations. The radiographic densities of each step of the step-wedge, and of carious and normal dentine of the phantom tooth, were determined by means of an RMI Digital Densitometer.(ABSTRACT TRUNCATED AT 250 WORDS)

Simultaneous Quantification of Gymnemic Acid as Gymnemagenin and Charantin as β-Sitosterol Using Validated HPTLC Densitometric Method.

PubMed

Ahamad, Javed; Amin, Saima; Mir, Showkat R

2015-08-01

Gymnemic acid and charantin are well-established antidiabetic phytosterols found in Gymnema sylvestre and Momordica charantia, respectively. The fact that these plants are often used together in antidiabetic poly-herbal formulations lured us to develop an HPTLC densitometric method for the simultaneous quantification of their bioactive compounds. Indirect estimation of gymnemic acid as gymnemagenin and charantin as β-sitosterol after hydrolysis has been proposed. Aluminum-backed silica gel 60 F254 plates (20 × 10 cm) were used as stationary phase and toluene-ethyl acetate-methanol-formic acid (60 : 20 : 15 : 5, v/v) as mobile phase. Developed chromatogram was scanned at 550 nm after derivatization with modified vanillin-sulfuric acid reagent. Regression analysis of the calibration data showed an excellent linear relationship between peak area versus concentration of the analytes. Linearity was found to be in the range of 500-2,500 and 100-500 ng/band for gymnemagenin and β-sitosterol, respectively. The suitability of the developed HPTLC method for simultaneous estimation of analytes was established by validating it as per the ICH guidelines. The limits of detection and quantification for gymnemagenin were found to be ≈60 and ≈190 ng/band, and those for β-sitosterol ≈30 and ≈90 ng/band, respectively. The developed method was found to be linear (r(2) = 0.9987 and 0.9943), precise (relative standard deviation <1.5 and <2% for intra- and interday precision) and accurate (mean recovery ranged between 98.43-101.44 and 98.68-100.20%) for gymnemagenin and β-sitosterol, respectively. The proposed method was also found specific and robust for quantification of both the analytes and was successfully applied to herbal drugs and in-house herbal formulation without any interference. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Validated Densitometric TLC-Method for the Simultaneous Analysis of (R)- and (S)-Citalopram and its Related Substances Using Macrocyclic Antibiotic as a Chiral Selector: Application to the Determination of Enantiomeric Purity of Escitalopram

PubMed Central

Soliman, Suzan Mahmoud

2012-01-01

A novel economic procedure for the simultaneous stereospecific separation and analysis of (R)- and (S)-citalopram and its related substances or impurities has been developed and validated. Chromatography was performed on silica gel 60 F254 plates with acetonitrile: methanol: water (15:2.5:2.5: v/v/v) as a mobile phase containing 1.5 mM norvancomycin or 2.5 mM vancomycin as a selector at ambient temperature. (R)- and (S)-citalopram enantiomers in presence of its related substances; citalopram citadiol and citalopram N-oxide were well separated with significant Rf values of 0.33 ± 0.02, 0.85 ± 0.02, 0.45 ± 0.02 and 0.22 ± 0.02, respectively. The spots were detected with either iodine vapor, or by use of a UV lamp followed by densitometric measurement at 239 nm. All variables affecting the resolution, such as concentration of chiral selectors, mobile phase system at different temperatures and pH-values were investigated and the conditions were optimized. Calibration plots for analysis of (R)- and (S)-enantiomers were linear in the range of 0.2-16.8 μg/10 μl (R≥0.9994, n=6) with acceptable precision (%RSD<2.0) and accuracy (99.70 ± 0.85% and 99.51 ± 0.61% for (S)-citalopram and escitalopram, respectively). The limit of detection and quantification were 0.08 μg/10 μl and 0.25 μg/10 μl, respectively, for (R)- and (S)-citalopram. The proposed method is simple, selective, and robust and can be applied for quantitative determination of enantiomeric purity of (R)- and (S)-citalopram (escitalopram) as well as the related impurities in drug substances and pharmaceutical preparations. The method can be useful to investigate adulteration of pure isomer with the cheep racemic form. PMID:23675256

Cross-reactivity of anti-chicken IgY antibody with immunoglobulins of exotic avian species.

PubMed

Cray, Carolyn; Villar, David

2008-09-01

A major challenge in the serologic diagnosis of infectious diseases in exotic birds is the limited availability of species-specific antibodies. The purpose of the current study was to determine if there is cross reactivity between commercially available anti-chicken IgY antibodies and immunoglobulins of several avian species, with particular emphasis on psittacines. To quantitate the reactivity with anti-chicken IgY, Western blot analysis was performed using plasma samples from many different avian species. Results were compared with gamma globulin fraction quantitation obtained by protein electrophoresis. By Western blot, 2 protein bands corresponding to the heavy and light chains of chicken IgY were identified in species from 21 avian orders using 1 of 2 rabbit anti-chicken IgY antibodies. Densitometric analysis showed that the amount of immunoglobulin estimated from Western blots correlated strongly with data from protein electrophoresis assays. The results demonstrate that some commercially available anti-chicken IgY antibodies exhibit good cross-reactivity with most avian species.

Characteristics of Kodak Insight, an F-speed intraoral film.

PubMed

Ludlow, J B; Platin, E; Mol, A

2001-01-01

This study reports film speed, contrast, exposure latitude, resolution, and response to processing solution depletion of Kodak Insight intraoral film. Densitometric curves were generated by using International Standards Organization protocol. Additional curves were generated for Ultra-speed, Ektaspeed Plus, and Insight films developed in progressively depleted processing solutions. Eight observers viewed images of a resolution test tool for maximum resolution assessment. Images of an aluminum step-wedge were reviewed to determine useful exposure latitude. Insight's sensitivity in fresh automatic processor solutions places it in the F-speed group. An average gradient of 1.8 was found with all film types. Insight provided 93% of the useful exposure latitude of Ektaspeed Plus film. Insight maintained contrast in progressively depleted processing solutions. Like Ektaspeed Plus, Insight was able to resolve at least 20 line-pairs per millimeter. Under International Standards Organization conditions, Insight required only 77% of the exposure of Ektaspeed Plus film. Insight film provided stable contrast in depleted processing solutions.

A duplicated PLP gene causing Pelizaeus-Merzbacher disease detected by comparative multiplex PCR

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

Inoue, K.; Sugiyama, N.; Kawanishi, C.

1996-07-01

Pelizaeus-Merzbacher disease (PMD) is an X-linked dysmyelinating disorder caused by abnormalities in the proteolipid protein (PLP) gene, which is essential for oligodendrocyte differentiation and CNS myelin formation. Although linkage analysis has shown the homogeneity at the PLP locus in patients with PMD, exonic mutations in the PLP gene have been identified in only 10% - 25% of all cases, which suggests the presence of other genetic aberrations, including gene duplication. In this study, we examined five families with PMD not carrying exonic mutations in PLP gene, using comparative multiplex PCR (CM-PCR) as a semiquantitative assay of gene dosage. PLP genemore » duplications were identified in four families by CM-PCR and confirmed in three families by densitometric RFLP analysis. Because a homologous myelin protein gene, PMP22, is duplicated in the majority of patients with Charcot-Marie-Tooth 1A, PLP gene overdosage may be an important genetic abnormality in PMD and affect myelin formation. 38 ref., 5 figs., 2 tabs.« less

HPTLC determination of diosgenin in fenugreek seeds.

PubMed

Król-Kogus, Barbara; Lamine, Khenifi Mohammed; Migas, Piotr; Boudjeniba, Messaoud; Krauze-Baranowska, Mirosława

2018-03-01

A new HPTLC-densitometric method for diosgenin determination in fenugreek seeds was established after optimization of the conditions for efficient saponin extraction and acid hydrolysis. Several procedures were tested, the best of which was a three-step Soxhlet extraction, followed by hydrolysis of the obtained methanolic extract with 2 mol L-1 H2SO4. Best diosgenin separation from other hydrolysis products was obtained on HPTLC Si60F254 plates u sing a mixture of n-heptane/ethyl acetate (7:3, V/V) and modified anisaldehyde as a spraying reagent. The method was preliminarily validated and the determined amounts of diosgenin in fenugreek seeds of Polish and African origin were found to be similar and ranged from 0.12-0.18 %.

Modulatory influence of juvenile hormone analogue (JHa) and 20-hydroxyecdysone on lipophorin synthesis in red cotton bug, Dysdercus cingulatus Fabr.

PubMed

Mohan, K G; Muraleedharan, D

2005-12-01

Topical supply of methoprene, a juvenile hormone analogue (JHa) caused notable morphological disturbance in insects. Topical supply of methoprene to newly emerged adult female D. cingulatus caused notable disturbance and induced a dramatic reduction in the total haemolymph protein pattern and lipophorin production in tissues like fat body, ovary and haemolymph. Total protein concentration in haemolymph also showed significant reduction in 1 day old insects but increased slightly as age advanced. Application of 20-hydroxyecdysone (20-HE) to 2-day-old adult female stimulated protein synthesis intensively. Lipophorin levels in fat body and ovary also simultaneously increased. Densitometric analysis revealed that methoprene inhibits while 20-HE stimulates lipophorin production in D. cingulatus.

Development of a simplified densitometer for the determination of aflatoxins by thin-layer chromatography.

PubMed

Stroka, J; Anklam, E

2000-12-29

A simple, miniaturised and low power consuming (battery, fully semiconductor based) detector cell (SeBaDeC) was developed for the densitometric measurement of aflatoxins on TLC plates. A UV-light emitting diode (UV-LED) with a peak emission wavelength of 370 nm was used for fluorescence excitation, while a photo diode with a peak sensitivity of 440 nm in combination with a 418 nm cut-off filter was applied for detecting the fluorescence intensity. The resulting signal was further amplified by means of a commonly used operational amplifier integrated circuit (OA) and directly converted into a digital signal with a simple analogue-digital-converter (ADC). This signal was recorded at the serial (RS232) port of a portable PC and processed with a spreadsheet program. The software used for data recording is freeware and available in its source code, and the long lifetime of the UV-LED (up to 10 000 h) permits a maintenance free application of this device. This simplified device has shown to be able to detect concentrations of aflatoxins of 1 ng, thus offering a cheap and sensitive alternative to currently available TCL scanners.

Nuclear morphology for the detection of alterations in bronchial cells from lung cancer: an attempt to improve sensitivity and specificity.

PubMed

Fafin-Lefevre, Mélanie; Morlais, Fabrice; Guittet, Lydia; Clin, Bénédicte; Launoy, Guy; Galateau-Sallé, Françoise; Plancoulaine, Benoît; Herlin, Paulette; Letourneux, Marc

2011-08-01

To identify which morphologic or densitometric parameters are modified in cell nuclei from bronchopulmonary cancer based on 18 parameters involving shape, intensity, chromatin, texture, and DNA content and develop a bronchopulmonary cancer screening method relying on analysis of sputum sample cell nuclei. A total of 25 sputum samples from controls and 22 bronchial aspiration samples from patients presenting with bronchopulmonary cancer who were professionally exposed to cancer were used. After Feulgen staining, 18 morphologic and DNA content parameters were measured on cell nuclei, via image cytom- etry. A method was developed for analyzing distribution quantiles, compared with simply interpreting mean values, to characterize morphologic modifications in cell nuclei. Distribution analysis of parameters enabled us to distinguish 13 of 18 parameters that demonstrated significant differences between controls and cancer cases. These parameters, used alone, enabled us to distinguish two population types, with both sensitivity and specificity > 70%. Three parameters offered 100% sensitivity and specificity. When mean values offered high sensitivity and specificity, comparable or higher sensitivity and specificity values were observed for at least one of the corresponding quantiles. Analysis of modification in morphologic parameters via distribution analysis proved promising for screening bronchopulmonary cancer from sputum.

Computed tomographic-based quantification of emphysema and correlation to pulmonary function and mechanics.

PubMed

Washko, George R; Criner, Gerald J; Mohsenifar, Zab; Sciurba, Frank C; Sharafkhaneh, Amir; Make, Barry J; Hoffman, Eric A; Reilly, John J

2008-06-01

Computed tomographic based indices of emphysematous lung destruction may highlight differences in disease pathogenesis and further enable the classification of subjects with Chronic Obstructive Pulmonary Disease. While there are multiple techniques that can be utilized for such radiographic analysis, there is very little published information comparing the performance of these methods in a clinical case series. Our objective was to examine several quantitative and semi-quantitative methods for the assessment of the burden of emphysema apparent on computed tomographic scans and compare their ability to predict lung mechanics and function. Automated densitometric analysis was performed on 1094 computed tomographic scans collected upon enrollment into the National Emphysema Treatment Trial. Trained radiologists performed an additional visual grading of emphysema on high resolution CT scans. Full pulmonary function test results were available for correlation, with a subset of subjects having additional measurements of lung static recoil. There was a wide range of emphysematous lung destruction apparent on the CT scans and univariate correlations to measures of lung function were of modest strength. No single method of CT scan analysis clearly outperformed the rest of the group. Quantification of the burden of emphysematous lung destruction apparent on CT scan is a weak predictor of lung function and mechanics in severe COPD with no uniformly superior method found to perform this analysis. The CT based quantification of emphysema may augment pulmonary function testing in the characterization of COPD by providing complementary phenotypic information.

Application of Scion image software to the simultaneous determination of curcuminoids in turmeric (Curcuma longa).

PubMed

Sotanaphun, Uthai; Phattanawasin, Panadda; Sriphong, Lawan

2009-01-01

Curcumin, desmethoxycurcumin and bisdesmethoxycurcumin are bioactive constituents of turmeric (Curcuma longa). Owing to their different potency, quality control of turmeric based on the content of each curcuminoid is more reliable than that based on total curcuminoids. However, to perform such an assay, high-cost instrument is needed. To develop a simple and low-cost method for the simultaneous quantification of three curcuminoids in turmeric using TLC and the public-domain software Scion Image. The image of a TLC chromatogram of turmeric extract was recorded using a digital scanner. The density of the TLC spot of each curcuminoid was analysed by the Scion Image software. The density value was transformed to concentration by comparison with the calibration curve of standard curcuminoids developed on the same TLC plate. The polynomial regression data for all curcuminoids showed good linear relationship with R(2) > 0.99 in the concentration range of 0.375-6 microg/spot. The limits of detection and quantitation were 43-73 and 143-242 ng/spot, respectively. The method gave adequate precision, accuracy and recovery. The contents of each curcuminoid determined using this method were not significantly different from those determined using the TLC densitometric method. TLC image analysis using Scion Image is shown to be a reliable method for the simultaneous analysis of the content of each curcuminoid in turmeric.

Oral treatment with retinoic acid decreases bone mass in rats.

PubMed

Hotchkiss, Charlotte E; Latendresse, John; Ferguson, Sherry A

2006-12-01

13-cis-retinoic acid (13-cis-RA, isotretinoin) is used to treat severe recalcitrant acne. Other retinoids have adverse effects on bone. Recent studies of human patients treated with 13-cis-RA have had varying results, perhaps because of variability among patients and the lack of control groups. The effects of retinoids have been studied in rodents, but little information is available regarding the effects of clinically relevant retinoid doses as evaluated by use of bone densitometric techniques. We treated rats for 15 or 20 wk with 13-cis-RA, all-trans-RA, or soybean oil (control) by gavage. We used dual-energy X-ray absorptiometry, histomorphometry, and histologic evaluation to evaluate effects on bone. Spontaneous long bone fractures occurred in some rats treated with 15 mg/kg all-trans-RA daily. Bone mineral density, bone mineral content, bone diameter, and cortical thickness of the femur were reduced in rats treated daily with 10 or 15 mg/kg all-trans-RA or 30 mg/kg 13-cis-RA. The lumbar spine was not affected. Although the effects of 13-cis-RA were not as dramatic as those of all-trans-RA, further study of the effects of 13-cis-RA on long bones is warranted.

Ibandronate and cementless total hip arthroplasty: densitometric measurement of periprosthetic bone mass and new therapeutic approach to the prevention of aseptic loosening

PubMed Central

Muratore, Maurizio; Quarta, Eugenio; Quarta, Laura; Calcagnile, Fabio; Grimaldi, Antonella; Orgiani, M. Antonio; Marsilio, Antonio; Rollo, Giuseppe

2012-01-01

Summary Studies of the mechanisms of periprosthetic bone loss have led to the development of pharmacologic strategies intended to enhance bone mass recovery after surgery and consequently prevent aseptic loosening and prolong the implant survival. Bisphosphonates, potent anti-resorptive drugs widely used in the treatment of osteoporosis and other disorders of bone metabolism, were shown to be particularly effective in reducing periprosthetic bone resorption in the first year after hip and knee arthroplasty, both cemented and cementless. Based on these results, we investigated the inhibitory effects of ibandronate on periprosthetic bone loss in a 2-year study of postmenopausal women that underwent cementless total hip arthroplasty. In the first 6 months both groups (A, treated with ibandronate 3 mg i.v. within five days after surgery and then with oral ibandronate 150 mg/month, plus calcium and vitamin D supplementation; and B, treated with calcium and vitamin D supplementation only) experienced bone loss, though to a lesser extent in group A. After 12 months, group A showed a remarkable BMD recovery, that was statistically significant versus baseline values (about +1, 74% of global BMD) and most evident in region R1 (+3, 81%) and R2 (+4, 12%); in group B, on the contrary, BMD values were unchanged compared with those at 6 months post-surgery. Quality of life scores also showed a greater improvement in group A, both at 6 and 12 months after surgery, likely because of the pain-reducing effects of ibandronate treatment. PMID:22783337

[Promoter hypermethylation status of the mismatch repair gene hMLH1 in patients with sporadic renal cell carcinoma].

PubMed

Salinas-Sánchez, Antonio S; Rubio-del-Campo, Antonio; Sánchez-Sánchez, Francisco; Giménez-Bachs, José M; Donate-Moreno, María J; García-Olmo, Dolores C; Escribano-Martínez, Julio

2006-04-01

Epigenetic inactivation is a gene function abnormality that produces no changes in the DNA sequence, with the most frequent epigenetic alteration being hypermethylation of CpG islands in the promoter regions of the genes. Based on recent indications of a potential relationship between mismatch repair genes and renal cell carcinoma (RCC), we were interested in investigating the existence of promoter hypermethylation of the hMLH1 gene in tumor DNA samples from patients with sporadic RCC. Sixty-five tumor tissue specimens were collected consecutively. The DNA was first obtained and purified, then digested with the restriction enzymes Hpa II and Msp I, followed by polimerase chain reaction amplification of 3 promoter regions of the hMLH1 gene, agarose gel electrophoresis, and densitometric analysis of the images of the amplified bands. Mean patient age was 63.7 years. The most frequent cell type was clear cell carcinoma (67.7%). 73.9% of tumors were diagnosed in stages below pT2, 9.3% had gland involvement and 20%, distant metastasis. No somatic hypermethylation was detected in the promoter region of the hMLH1 gene in any of the patients studied. Our data indicate that promoter hypermethylation of the hMLH1 gene is not implicated in the pathogenesis of sporadic RCC, and therefore the existence of another type of mutation, microsatellite instability and/or loss of heterozygosity should be examined to determine the possible role of this gene in sporadic RCC.

Comparison of four software packages for CT lung volumetry in healthy individuals.

PubMed

Nemec, Stefan F; Molinari, Francesco; Dufresne, Valerie; Gosset, Natacha; Silva, Mario; Bankier, Alexander A

2015-06-01

To compare CT lung volumetry (CTLV) measurements provided by different software packages, and to provide normative data for lung densitometric measurements in healthy individuals. This retrospective study included 51 chest CTs of 17 volunteers (eight men and nine women; mean age, 30 ± 6 years), who underwent spirometrically monitored CT at total lung capacity (TLC), functional residual capacity (FRC), and mean inspiratory capacity (MIC). Volumetric differences assessed by four commercial software packages were compared with analysis of variance (ANOVA) for repeated measurements and benchmarked against the threshold for acceptable variability between spirometric measurements. Mean lung density (MLD) and parenchymal heterogeneity (MLD-SD) were also compared with ANOVA. Volumetric differences ranged from 12 to 213 ml (0.20 % to 6.45 %). Although 16/18 comparisons (among four software packages at TLC, MIC, and FRC) were statistically significant (P < 0.001 to P = 0.004), only 3/18 comparisons, one at MIC and two at FRC, exceeded the spirometry variability threshold. MLD and MLD-SD significantly increased with decreasing volumes, and were significantly larger in lower compared to upper lobes (P < 0.001). Lung volumetric differences provided by different software packages are small. These differences should not be interpreted based on statistical significance alone, but together with absolute volumetric differences. • Volumetric differences, assessed by different CTLV software, are small but statistically significant. • Volumetric differences are smaller at TLC than at MIC and FRC. • Volumetric differences rarely exceed spirometric repeatability thresholds at MIC and FRC. • Differences between CTLV measurements should be interpreted based on comparison of absolute differences. • MLD increases with decreasing volumes, and is larger in lower compared to upper lobes.

Quantification of sunscreen ethylhexyl triazone in topical skin-care products by normal-phase TLC/densitometry.

PubMed

Sobanska, Anna W; Pyzowski, Jaroslaw

2012-01-01

Ethylhexyl triazone (ET) was separated from other sunscreens such as avobenzone, octocrylene, octyl methoxycinnamate, and diethylamino hydroxybenzoyl hexyl benzoate and from parabens by normal-phase HPTLC on silica gel 60 as stationary phase. Two mobile phases were particularly effective: (A) cyclohexane-diethyl ether 1 : 1 (v/v) and (B) cyclohexane-diethyl ether-acetone 15 : 1 : 2 (v/v/v) since apart from ET analysis they facilitated separation and quantification of other sunscreens present in the formulations. Densitometric scanning was performed at 300 nm. Calibration curves for ET were nonlinear (second-degree polynomials), with R > 0.998. For both mobile phases limits of detection (LOD) were 0.03 and limits of quantification (LOQ) 0.1 μg spot(-1). Both methods were validated.

Experiment K-6-18. Study of muscarinic and gaba (benzodiazepine) receptors in the sensory-motor cortex, hippcampus and spinal code

NASA Technical Reports Server (NTRS)

Daunton, N.; Damelio, F.; Krasnov, I.

1990-01-01

Frontal lobe samples of rat brains flown aboard Cosmos 1887 were processed for the study of muscarinic (cholinergic) and GABA (benzodiazepine) receptors and for immunocytochemical localization of the neurotransmitter gamma-aminobutyric acid (GABA) and glial fibrillary acidic protein (GFAP). Although radioactive labeling of both muscarinic cholinergic and GABA (benzodiazepine) receptors proved to be successful with the techniques employed, distinct receptor localization of individual laminae of the frontal neocortex was not possible since the sampling of the area was different in the various groups of animals. In spite of efforts made for proper orientation and regional identification of laminae, it was found that a densitometric (quantitation of autoradiograms) analysis of the tissue did not contribute to the final interpretation of the effects of weightlessness on these receptors. As to the immunocytochemical studies the use of both markers, GFAP and GABA antiserum, confirmed the suitability of the techniques for use in frozen material. However, similar problems to those encountered in the receptor studies prevented an adequate interpretation of the effects of micro-G exposure on the localization and distribution of GABA and GFAP. This study did, however, confirm the feasibility of investigating neurotransmitters and their receptors in future space flight experiments.

Propolis Modifies Collagen Types I and III Accumulation in the Matrix of Burnt Tissue.

PubMed

Olczyk, Pawel; Wisowski, Grzegorz; Komosinska-Vassev, Katarzyna; Stojko, Jerzy; Klimek, Katarzyna; Olczyk, Monika; Kozma, Ewa M

2013-01-01

Wound healing represents an interactive process which requires highly organized activity of various cells, synthesizing cytokines, growth factors, and collagen. Collagen types I and III, serving as structural and regulatory molecules, play pivotal roles during wound healing. The aim of this study was to compare the propolis and silver sulfadiazine therapeutic efficacy throughout the quantitative and qualitative assessment of collagen types I and III accumulation in the matrix of burnt tissues. Burn wounds were inflicted on pigs, chosen for the evaluation of wound repair because of many similarities between pig and human skin. Isolated collagen types I and III were estimated by the surface plasmon resonance method with a subsequent collagenous quantification using electrophoretic and densitometric analyses. Propolis burn treatment led to enhanced collagens and its components expression, especially during the initial stage of the study. Less expressed changes were observed after silver sulfadiazine (AgSD) application. AgSD and, with a smaller intensity, propolis stimulated accumulation of collagenous degradation products. The assessed propolis therapeutic efficacy, throughout quantitatively and qualitatively analyses of collagen types I and III expression and degradation in wounds matrix, may indicate that apitherapeutic agent can generate favorable biochemical environment supporting reepithelization.

Visualization of newt aragonitic otoconial matrices using transmission electron microscopy

NASA Technical Reports Server (NTRS)

Steyger, P. S.; Wiederhold, M. L.

1995-01-01

Otoconia are calcified protein matrices within the gravity-sensing organs of the vertebrate vestibular system. These protein matrices are thought to originate from the supporting or hair cells in the macula during development. Previous studies of mammalian calcitic, barrel-shaped otoconia revealed an organized protein matrix consisting of a thin peripheral layer, a well-defined organic core and a flocculent matrix inbetween. No studies have reported the microscopic organization of the aragonitic otoconial matrix, despite its protein characterization. Pote et al. (1993b) used densitometric methods and inferred that prismatic (aragonitic) otoconia have a peripheral protein distribution, compared to that described for the barrel-shaped, calcitic otoconia of birds, mammals, and the amphibian utricle. By using tannic acid as a negative stain, we observed three kinds of organic matrices in preparations of fixed, decalcified saccular otoconia from the adult newt: (1) fusiform shapes with a homogenous electron-dense matrix; (2) singular and multiple strands of matrix; and (3) more significantly, prismatic shapes outlined by a peripheral organic matrix. These prismatic shapes remain following removal of the gelatinous matrix, revealing an internal array of organic matter. We conclude that prismatic otoconia have a largely peripheral otoconial matrix, as inferred by densitometry.

Antimicrobial activity of silica coated silicon nano-tubes (SCSNT) and silica coated silicon nano-particles (SCSNP) synthesized by gas phase condensation.

PubMed

Tank, Chiti; Raman, Sujatha; Karan, Sujoy; Gosavi, Suresh; Lalla, Niranjan P; Sathe, Vasant; Berndt, Richard; Gade, W N; Bhoraskar, S V; Mathe, Vikas L

2013-06-01

Silica-coated, silicon nanotubes (SCSNTs) and silica-coated, silicon nanoparticles (SCSNPs) have been synthesized by catalyst-free single-step gas phase condensation using the arc plasma process. Transmission electron microscopy and scanning tunneling microscopy showed that SCSNTs exhibited a wall thickness of less than 1 nm, with an average diameter of 14 nm and a length of several 100 nm. Both nano-structures had a high specific surface area. The present study has demonstrated cheaper, resistance-free and effective antibacterial activity in silica-coated silicon nano-structures, each for two Gram-positive and Gram-negative bacteria. The minimum inhibitory concentration (MIC) was estimated, using the optical densitometric technique, and by determining colony-forming units. The MIC was found to range in the order of micrograms, which is comparable to the reported MIC of metal oxides for these bacteria. SCSNTs were found to be more effective in limiting the growth of multidrug-resistant Staphylococcus aureus over SCSNPs at 10 μg/ml (IC 50 = 100 μg/ml).

Optimizing Bone Health in Duchenne Muscular Dystrophy.

PubMed

Buckner, Jason L; Bowden, Sasigarn A; Mahan, John D

2015-01-01

Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder characterized by progressive muscle weakness, with eventual loss of ambulation and premature death. The approved therapy with corticosteroids improves muscle strength, prolongs ambulation, and maintains pulmonary function. However, the osteoporotic impact of chronic corticosteroid use further impairs the underlying reduced bone mass seen in DMD, leading to increased fragility fractures of long bones and vertebrae. These serious sequelae adversely affect quality of life and can impact survival. The current clinical issues relating to bone health and bone health screening methods in DMD are presented in this review. Diagnostic studies, including biochemical markers of bone turnover and bone mineral density by dual energy X-ray absorptiometry (DXA), as well as spinal imaging using densitometric lateral spinal imaging, and treatment to optimize bone health in patients with DMD are discussed. Treatment with bisphosphonates offers a method to increase bone mass in these children; oral and intravenous bisphosphonates have been used successfully although treatment is typically reserved for children with fractures and/or bone pain with low bone mass by DXA.

A volumetric pulmonary CT segmentation method with applications in emphysema assessment

NASA Astrophysics Data System (ADS)

Silva, José Silvestre; Silva, Augusto; Santos, Beatriz S.

2006-03-01

A segmentation method is a mandatory pre-processing step in many automated or semi-automated analysis tasks such as region identification and densitometric analysis, or even for 3D visualization purposes. In this work we present a fully automated volumetric pulmonary segmentation algorithm based on intensity discrimination and morphologic procedures. Our method first identifies the trachea as well as primary bronchi and then the pulmonary region is identified by applying a threshold and morphologic operations. When both lungs are in contact, additional procedures are performed to obtain two separated lung volumes. To evaluate the performance of the method, we compared contours extracted from 3D lung surfaces with reference contours, using several figures of merit. Results show that the worst case generally occurs at the middle sections of high resolution CT exams, due the presence of aerial and vascular structures. Nevertheless, the average error is inferior to the average error associated with radiologist inter-observer variability, which suggests that our method produces lung contours similar to those drawn by radiologists. The information created by our segmentation algorithm is used by an identification and representation method in pulmonary emphysema that also classifies emphysema according to its severity degree. Two clinically proved thresholds are applied which identify regions with severe emphysema, and with highly severe emphysema. Based on this thresholding strategy, an application for volumetric emphysema assessment was developed offering new display paradigms concerning the visualization of classification results. This framework is easily extendable to accommodate other classifiers namely those related with texture based segmentation as it is often the case with interstitial diseases.

Stress and transcriptional regulation of tick ferritin HC.

PubMed

Mulenga, A; Simser, J A; Macaluso, K R; Azad, A F

2004-08-01

We previously identified a partial Dermacentor variabilis cDNA encoding ferritin HC (HC) subunit homolog (DVFER) that was differentially upregulated in Rickettsia montanensis infected ticks (Mulenga et al., 2003a). We have used rapid amplification of cDNA ends to clone full-length DVFER cDNA and its apparent ortholog from the wood tick, D. andersoni (DAFER), both of which show high sequence similarity to vertebrate than insect ferritin. Both DVFER and DAFER contain the stem-loop structure of a putative iron responsive element in the 5' untranslated region (nucleotide positions, 16-42) and the feroxidase centre loop typical for vertebrate ferritin HC subunits. Quantitative Western and Northern blotting analyses of protein and RNA from unfed and partially fed whole tick as well as dissected tick tissues demonstrated that DVFER is constitutively and ubiquitously expressed. Based on densitometric analysis of detected protein and mRNA bands, DVFER is predominantly expressed in the midgut, and to a lesser extent in the salivary glands, ovary and fatbody. Sham treatment (mechanical injury) and Escherichia coli challenge of D. variabilis ticks stimulated statistically significant (approximately 1.5- and approximately 3.0-fold, respectively) increases in DVFER mRNA abundance over time point matched naive control ticks. These data suggest that DVFER mRNA is nonspecifically up regulated in response to mechanical injury or bacterial infection induced stress.

Poverty is a risk factor for osteoporotic fractures.

PubMed

Navarro, M C; Sosa, M; Saavedra, P; Lainez, P; Marrero, M; Torres, M; Medina, C D

2009-03-01

This study assesses the possible association between poverty and osteoporosis and/or fragility fractures in a population of postmenopausal women. We found that postmenopausal women with low socioeconomic status had lower values of BMD at the lumbar spine, a higher prevalence of densitometric osteoporosis, and a higher prevalence of total and vertebral fractures. Some lifestyles are related to the presence of osteoporosis and/or fragility fractures, whereas poverty is related to some lifestyles. Because of this, we studied the possible association of poverty with osteoporosis and fractures. This was an observational, cross-sectional study performed in the Canary Islands, Spain. Participants consisted of a total of 1,139 ambulatory postmenopausal women aged 50 years or older with no previous osteoporosis diagnosis and who were enrolled in some epidemiological studies. The prevalence of fractures (vertebral and non-vertebral) and the prevalence of osteoporosis (T-score <-2.5 either at the lumbar spine or the femoral neck). A previously validated questionnaire elicited the most important risk factors for osteoporosis: socioeconomic status, defined by the annual income was also assessed by a personal interview. A dorso-lateral X-ray of the spine was performed, and bone mineral density (BMD) was measured by DXA in the lumbar spine (L2-L4) and proximal femur. Compared to women with a medium and high socioeconomic status (n = 665), those who were classified into poverty (annual family income lower than 6,346.80 Euros, in a one-member family, n = 474), were older and heavier and had lower height, lower prevalence of tobacco and alcohol consumption, lower use of HRT and higher use of thiazides. After correcting for age and body mass index (BMI), women in poverty had lower spine BMD values than women with a medium and high socioeconomic status (0.840 g/cm(2) vs. 0.867 g/cm(2), p = 0.005), but there were no statistical differences in femoral neck BMD between groups. The prevalence of osteoporosis was also higher in women in poverty [40.6% vs. 35.6%, (OR 1.35, CI 95%: 1.03; 1.76)] after adjusting by age and BMI. Moreover, 37.8% of women in poverty had a history of at least one fragility fracture compared to 27.7% of women not in poverty (OR: 1.45, CI 95%: 1.11; 1.90). The prevalence of vertebral fractures was also higher in women in poverty 24.7% vs. 13.4%, (OR 2.01, CI 95%: 1.44; 2.81). Postmenopausal women with low socioeconomic status had lower values of BMD at the lumbar spine, and a higher prevalence of densitometric osteoporosis, and a higher prevalence of total and vertebral fractures. Because of this, apart from the well known risk factors for osteoporosis, poverty should be taken into account as a possible risk factor for both osteoporosis and fragility fractures, in order to establish sanitary strategies to protect unfavoured postmenopausal women.

Sera of IgA nephropathy patients contain a heterogeneous population of relatively cationic alpha-heavy chains.

PubMed

Shuib, A S; Chua, C T; Hashim, O H

1998-01-01

Sera of IgA nephropathy (IgAN) patients and normal subjects were analysed by two-dimensional (2-D) gel electrophoresis. Densitometric analysis of the 2-D gels of IgAN patients and normal subjects revealed that their protein maps were comparable. There was no shift of pI values in the major alpha-heavy chain spots. However, the volume of the alpha-heavy chain bands were differently distributed. Distribution was significantly lower at the anionic region in IgAN patients (mean anionic:cationic ratio of 1.184 +/- 0.311) as compared to normal healthy controls (mean anionic:cationic ratio of 2.139 +/- 0.538). Our data are in support of the previously reported findings that IgA1 of IgAN patients were lacking in sialic acid residues.

Quantification of Sunscreen Ethylhexyl Triazone in Topical Skin-Care Products by Normal-Phase TLC/Densitometry

PubMed Central

Sobanska, Anna W.; Pyzowski, Jaroslaw

2012-01-01

Ethylhexyl triazone (ET) was separated from other sunscreens such as avobenzone, octocrylene, octyl methoxycinnamate, and diethylamino hydroxybenzoyl hexyl benzoate and from parabens by normal-phase HPTLC on silica gel 60 as stationary phase. Two mobile phases were particularly effective: (A) cyclohexane-diethyl ether 1 : 1 (v/v) and (B) cyclohexane-diethyl ether-acetone 15 : 1 : 2 (v/v/v) since apart from ET analysis they facilitated separation and quantification of other sunscreens present in the formulations. Densitometric scanning was performed at 300 nm. Calibration curves for ET were nonlinear (second-degree polynomials), with R > 0.998. For both mobile phases limits of detection (LOD) were 0.03 and limits of quantification (LOQ) 0.1 μg spot−1. Both methods were validated. PMID:22629203

Regional distribution of neuropeptide Y mRNA in postmortem human brain.

PubMed

Brené, S; Lindefors, N; Kopp, J; Sedvall, G; Persson, H

1989-12-01

The distribution of messenger RNA encoding neuropeptide Y (NPY) was studied in 11 different postmortem human brain regions using in situ hybridization histochemistry, and RNA blot analysis. In situ hybridization data revealed that the highest numerical density of labeled cells corresponded to neurons in accumbens area, caudate nucleus, putamen, and substantia innominata. Significantly fewer NPY mRNA-containing neurons were found in frontal and parietal cortex, amygdaloid body and dentate gyrus. No NPY mRNA-containing cells were found in substantia nigra. NPY mRNA-positive neurons from all regions studied showed relatively similar labeling, as revealed by computerized image analysis. Blot analysis showed an approximately 0.8 kb NPY mRNA in all brain regions studied, except in substantia nigra and cerebellum. Densitometric scanning of the autoradiograms revealed levels of NPY mRNA in the following order: putamen greater than caudate nucleus greater than frontal cortex (Brodmann areas 4 and 6) greater than temporal cortex (Brodmann area 38) greater than parietal cortex (Brodmann areas 5 and 7) greater than frontal cortex (Brodmann area 11). Hence, although NPY mRNA is widely distributed in neurons of the human brain large regional variation exists, with the highest expression in accumbens area and parts of the basal ganglia.

Dual-energy computed tomography for the detection of focal liver lesions.

PubMed

Lago, K N; Vallejos, J; Capuñay, C; Salas, E; Reynoso, E; Carpio, J B; Carrascosa, P M

To qualitatively and quantitatively explore the spectral study of focal liver lesions, comparing it with the usual polychromatic assessment with single-energy computed tomography. We prospectively studied 50 patients with at least one focal liver lesion who were referred for abdominal multidetector computed tomography with intravenous contrast material. The portal phase was acquired with dual energy sources. The density of the lesions and of the surrounding liver parenchyma was measured both in the baseline polychromatic acquisition and in the posterior monochromatic reconstructions at 40 keV, 70 keV, and 140 keV. Spectral curves were traced and the dual-energy indices and contrast-to-noise ratio were calculated. Lastly, the quality of the images and the detectability of the lesions were assessed qualitatively. Densitometric differences between the different types of lesions (avascular and vascularized) and the liver were greater at low energy levels (left side of the spectral curve) than in the polychromatic evaluation. In the subjective assessment, the 40keV energy level had the greatest lesion detectability. Monochromatic spectral study with dual-energy computed tomography provides better lesion detectability at 40keV compared to that provided by the ordinary polychromatic evaluation. Copyright © 2017 SERAM. Publicado por Elsevier España, S.L.U. All rights reserved.

Identification of vertebral deformities in the Polish population by morphometric X-ray absorptiometry - results of the EPOLOS study.

PubMed

Skowrońska-Jóźwiak, Elzbieta; Płudowski, Paweł; Karczmarewicz, Elzbieta; Lorenc, Roman; Lewiński, Andrzej

2009-01-01

The aim of the study was the determination of the prevalence of asymptomatic vertebral deformities in healthy persons of the Polish population, based on morphometric X-ray absorptiometry (MXA), and comparison of the results with data from literature, obtained by other techniques. The study involved 829 persons, including 520 women and 309 men, aged 18-79 years, untreated for osteoporosis before. The Th(4) to L(4) vertebrae were examined. Lateral scans of the thoracic-lumbar spine were made by an Expert-XL densitometer. Six point digitization was used to calculate the anterior (Ha), central (Hc), and posterior (Hp) height of the Th(4)-L(4) vertebral bodies. The vertebrae were defined as having prevalent deformities when at least one ratio value (Ha/Hp, Hc/Hp, Hp/Hp up, or Hp/Hp low) fell 3 SDs below or even more than the reference mean of that ratio at any vertebral level. The analysis was performed on 9629 vertebrae, of which 167 (1.75%), evaluated as deformed and considered as fractures, were observed in 113 patients (13.63 % of the examined patients). In 81 persons (74% of the patients with fractures; 9.7% of the studied population), single fractures were demonstrated, while in 28 persons, multiple deformities prevailed. Fractures occurred in 108 women (20.7% of the examined women) and 42 men (13.5% of the examined men). The highest incidence of deformities was observed in women over 55 years of age. First-degree deformities dominated. Deformities of the Th(8) and Th(6) vertebrae were most frequently observed. 1. Using MXA, it was found that in the Polish population deformities of vertebrae are common, as was demonstrated in X-ray morphometric studies in the European Vertebral Observation Study (EVOS). 2. Densitometric morphometry, as a non-invasive technique, may become a useful tool in the diagnostics of vertebral fractures.

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

Landry, L.G.; Pell, E.J.

Plants exposed to ozone (O{sub 3}) exhibited symptoms of premature senescence, including early decline in quantity of rubisco. O{sub 3}-induced oxidation may cause changes in protein conformation of rubisco, resulting in enhanced proteolysis. To test this hypothesis, rubisco was purified from two hybrid clones of Populus maximowizii x trichocarpa, clones 388 and 245, and treated in vitro with O{sub 3} or air. Rubisco was then challenged with bromelain, papain, chymotrypsin, carboxypeptidase A, or endoproteinase Glu-C and percent degradation measured by SDS-PAGE and densitometric scanning of the gels. Degree of rubisco sensitivity to oxidation may be related to available sulfhydryl (SH)more » groups on the protein. The number of SH groups in native and denatured rubisco was measured for purified rubisco of both clones by DTNB titration method. The relationship between sensitivity to proteolysis and number and availability of SH groups is discussed.« less

Effects of glucocorticoids on skeletal growth in rabbits evaluated by dual-photon absorptiometry, microscopic connectivity and vertebral compressive strength.

PubMed

Grardel, B; Sutter, B; Flautre, B; Viguier, E; Lavaste, F; Hardouin, P

1994-07-01

The effects of corticosteroid on bone were examined in female growing rabbits treated with 0.7 mg/kg per day prednisolone for 5 months. The evolution of whole-body total bone mineral measured by dual-photon absorptiometry showed a significant difference between the prednisolone-treated group and the control group from the first to the fifth month. The histomorphometric profile of corticosteroid-induced osteoporosis was observed, in particular the lower bone volume and thinner and fewer trabecular plates. Mechanical tests are possible on rabbit vertebrae and showed a very significant difference in bone strength between the prednisolone-treated and control groups, and a good correlation between mechanical tests and histomorphometric or densitometric results. This bone corticosteroid model shows that vertebral compression tests are possible on rabbit lumbar vertebrae. It may contribute to a better evaluation of corticosteroid treatments.

Crucial aspects of high performance thin layer chromatography quantitative validation. The case of determination of rosmarinic acid in different matrices.

PubMed

Coran, Silvia A; Mulas, Stefano; Mulinacci, Nadia

2012-01-13

A new HPTLC method was envisaged to determine rosmarinic acid (RA) in different matrices with the aim of testing the influence of optimizing the main HPTLC operative parameters in view of a more stringent validation process. HPTLC LiChrospher silica gel 60 F254s, 20 cm × 10 cm, plates with toluene:ethyl formate:formic acid (6:4:1, v/v) as the mobile phase were used. Densitometric determinations were performed in reflectance mode at 330 nm. The method was validated giving rise to a dependable and high throughput procedure well suited to routine applications. RA was quantified in the range of 132-660 ng with RSD of repeatability and intermediate precision not exceeding 2.0% and accuracy within the acceptance limits. The method was tested on several commercial preparations containing RA in different amounts. Copyright © 2011 Elsevier B.V. All rights reserved.

Putative melatonin receptors in a human biological clock

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

Reppert, S.M.; Weaver, D.R.; Rivkees, S.A.

In vitro autoradiography with /sup 125/I-labeled melatonin was used to examine melatonin binding sites in human hypothalamus. Specific /sup 125/I-labeled melatonin binding was localized to the suprachiasmatic nuclei, the site of a putative biological clock, and was not apparent in other hypothalamic regions. Specific /sup 125/I-labeled melatonin binding was consistently found in the suprachiasmatic nuclei of hypothalami from adults and fetuses. Densitometric analysis of competition experiments with varying concentrations of melatonin showed monophasic competition curves, with comparable half-maximal inhibition values for the suprachiasmatic nuclei of adults (150 picomolar) and fetuses (110 picomolar). Micromolar concentrations of the melatonin agonist 6-chloromelatonin completelymore » inhibited specific /sup 125/I-labeled melatonin binding, whereas the same concentrations of serotonin and norepinephrine caused only a partial reduction in specific binding. The results suggest that putative melatonin receptors are located in a human biological clock.« less

Enabling three-dimensional densitometric measurements using laboratory source X-ray micro-computed tomography

NASA Astrophysics Data System (ADS)

Pankhurst, M. J.; Fowler, R.; Courtois, L.; Nonni, S.; Zuddas, F.; Atwood, R. C.; Davis, G. R.; Lee, P. D.

2018-01-01

We present new software allowing significantly improved quantitative mapping of the three-dimensional density distribution of objects using laboratory source polychromatic X-rays via a beam characterisation approach (c.f. filtering or comparison to phantoms). One key advantage is that a precise representation of the specimen material is not required. The method exploits well-established, widely available, non-destructive and increasingly accessible laboratory-source X-ray tomography. Beam characterisation is performed in two stages: (1) projection data are collected through a range of known materials utilising a novel hardware design integrated into the rotation stage; and (2) a Python code optimises a spectral response model of the system. We provide hardware designs for use with a rotation stage able to be tilted, yet the concept is easily adaptable to virtually any laboratory system and sample, and implicitly corrects the image artefact known as beam hardening.

Monocarboxylate transporter 1 (MCT1) in the liver of pre-ruminant and adult bovines.

PubMed

Kirat, D; Inoue, H; Iwano, H; Yokota, H; Taniyama, H; Kato, S

2007-01-01

This study investigated the distribution and expression of monocarboxylate transporter 1 (MCT1) in the livers of pre-ruminant calves and adult bovines (bulls and cows), using different molecular biological techniques. Reverse transcription-polymerase chain reaction (RT-PCR) verified the presence of mRNA encoding for MCT1 in both pre-ruminant and adult bovine livers. Immunohistochemically, MCT1 was clearly demonstrated on the sinusoidal surfaces of bovine hepatocytes but its expression varied widely between pre-ruminants and adult bovines. In pre-ruminants, a faint hepatocellular expression of MCT1 was observed in a few hepatocytes, whereas an intense immunoreactive staining for MCT1 was shown in the majority of adult bovine hepatocytes. Western blot analysis also confirmed the results of the immunohistochemistry. Quantitative immunoblotting, as estimated by densitometric analysis, showed that the level of MCT1 in the liver of adult bovines was 8-9-fold greater (P<0.01) than that in pre-ruminant calf livers although no significant differences were detected between bulls and cows. The results demonstrated that MCT1 may play a crucial role in the transport of propionate in bovine liver, suggesting that MCT1 expression may be influenced by developmental and metabolic regulations.

Determination of molecular mass values of chondroitin sulfates by fluorophore-assisted carbohydrate electrophoresis (FACE).

PubMed

Buzzega, Dania; Maccari, Francesca; Volpi, Nicola

2010-03-11

Fluorophore-assisted carbohydrate electrophoresis (FACE) was applied to determine the molecular mass (M) values of various chondroitin sulfate (CS) samples. After labeling with 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS), FACE was able to resolve each CS sample as a discrete band depending on the M value. After densitometric acquisition, the migration distance of each CS standard was acquired and the third grade polynomial calibration standard curve was determined by plotting the logarithms of the M values as a function of migration ratio. Purified CS samples of different origin and the European Pharmacopeia CS standard were analyzed by both FACE and conventional high-performance size-exclusion liquid chromatography (HPSEC) methods. The molecular weight value on the top of the chromatographic peak (M(p)), the number-average M(n), weight-average M(w), and polydispersity (M(w)/M(n)) were examined by both techniques and found to be quite similar. This study demonstrates that FACE analysis is a suitable, sensitive and simple method for the determination of the M values of CS macromolecules with possible utilization in virtually any kind of research and development such as quality control laboratories. Copyright 2009 Elsevier B.V. All rights reserved.

Application of LANDSAT images to wetland study and land use classification in west Tennessee, part 1

NASA Technical Reports Server (NTRS)

Shahrokhi, F. (Principal Investigator); Jones, N. L.

1977-01-01

The author has identified the following significant results. densitometric analysis was performed on LANDSAT data to permit numerical classification of objects observed in the imagery on the basis of measurements of optical density. Relative light transmission measurements were taken on four types of scene elements in each of three LANDSAT black and white bands in order to determine which classification could be distinguished. The analysis of band 6 determined forest and agricultural classifications, but not the urban and wetlands. Both bands 4 and 5 showed a significant difference existed between the confirmed classification of wetlands-agriculture, and urban areas. Therefore, the combination of band 6 with either 4 or 5 would permit the separation of the urban from the wetland classification. To enhance the urban and wetland boundaries, the LANDSAT black and white bands were combined in a multispectral additive color viewer. Several combinations of filters and light intensities were used to obtain maximum discrimination between points of interest. The best results for enhancing wetland boundaries and urban areas were achieved by using a color composite (a blue, green, and red filter on bands 4, 5 and 6 respectively).

Alpha lipoic acid (ALA) modulates expression of apoptosis associated proteins in hippocampus of rats exposed during postnatal period to sodium arsenite (NaAsO2).

PubMed

Dixit, Shilpi; Dhar, Pushpa; Mehra, Raj D

2015-01-01

The present study focused on the role of exogenous alpha lipoic acid (ALA) in amelioration of inorganic arsenic ( iAs ) induced effects on apoptosis and apoptosis associated proteins in developing rat hippocampus. NaAsO 2 (1.5/2.0 mg/kg bw) alone or along with ALA (70 mg/kg bw) was administered to rat pups (experimental groups) by intraperitoneal (i.p.) route from postnatal day (PND) 4-15. Controls received no treatment/distilled water/ALA. On PND 16, the animals were perfusion fixed and the brains were processed for paraffin embedding (CV and TUNEL staining) and cryopreservation (immunohistochemistry). The fresh brain tissue was used for Western blotting. Significant increase was observed in TUNEL positive cells and Bax (pro-apoptotic protein) expression in hippocampal sub-regions of iAs alone treated groups, whereas Bcl-2 expression was intensified in animals receiving ALA with iAs . Densitometric analysis (Western blots) revealed optimal restoration of Bax and Bcl-2 ratio in animals receiving ALA with iAs , thereby suggesting the protective role of ALA in iAs induced developmental neurotoxicity.

Total lung capacity, residual volume and predicted residual volume in a densitometric study of older men.

PubMed Central

Latin, R W; Ruhling, R O

1986-01-01

Results of investigations using various lung volumes for hydrostatic weighing determinations (HWD) appear to be inconclusive. Often, these lung volumes are predicted and not clinically determined. For this reason, total lung capacity (TLC), a measured residual volume (RV), and a predicted residual volume (PRV) were used during HWDs to compare the techniques. Twenty-five older men, 56 to 70 years (means +/- 62.1 + 4.2 years) performed HWDs at RV (10 trials) and at TLC (3-5 trials). Values for body density and fat free mass were not significantly different between RV and TLC; both values were, however, significantly different from those derived using PRV. There were statistically significant differences (p less than 0.05) between all 3 per cent body fat values but the 1.1 per cent difference between TLC and RV may not be physiologically important. It was concluded that TLC and RV may be used comparably during HWDs, but a PRV may produce significantly different values. Since HWD at TLC is easily performed and circumvents the difficulties associated with the RV technique, it may be the preferred method for older subjects. PMID:3730758

Muscarinic receptor M1 and M2 subtypes in the human eye: QNB, pirenzipine, oxotremorine, and AFDX-116 in vitro autoradiography.

PubMed Central

Gupta, N; McAllister, R; Drance, S M; Rootman, J; Cynader, M S

1994-01-01

Muscarinic cholinergic agents are used to lower intraocular pressure in the medical management of glaucoma and subtypes of muscarinic receptors have now been recognised in many tissues including the eye. To localise muscarinic receptors and their M1 and M2 subtypes in the human eye, in vitro ligand binding and autoradiographic techniques with densitometric quantitation on postmortem eye sections were used. As ligands, [3H] quinuclydinyl benzylate (QNB) (non-subtype specific muscarinic antagonist), [3H]pirenzipine (M1 antagonist), [3H]oxotremorine (M2 muscarinic agonist), [3H]AFDX-116(11[(2[diethylaminomethyl]1-piperidinyl)acetyl]5 , 11dihydro-6H-pyrido [2,3b][1,4]benzodiazepine-6-one) (M2 antagonist) were studied. Specific binding sites for QNB, pirenzipine, and AFDX-116 were localised in the entire ciliary muscle, the iris, and ciliary epithelium. [3H]oxotremorine localised only in the longitudinal portion of the ciliary muscle, and additionally, was not localised in the iris or ciliary epithelium. These results suggest that oxotremorine, by binding selectively to receptors on the longitudinal ciliary muscle and inducing its contraction, may modulate outflow facility independently from accommodation and miosis. Images PMID:7918268

Radioactive Phosphorylation of Alcohols to Monitor Biocatalytic Diels-Alder Reactions

PubMed Central

Nierth, Alexander; Jäschke, Andres

2011-01-01

Nature has efficiently adopted phosphorylation for numerous biological key processes, spanning from cell signaling to energy storage and transmission. For the bioorganic chemist the number of possible ways to attach a single phosphate for radioactive labeling is surprisingly small. Here we describe a very simple and fast one-pot synthesis to phosphorylate an alcohol with phosphoric acid using trichloroacetonitrile as activating agent. Using this procedure, we efficiently attached the radioactive phosphorus isotope 32P to an anthracene diene, which is a substrate for the Diels-Alderase ribozyme—an RNA sequence that catalyzes the eponymous reaction. We used the 32P-substrate for the measurement of RNA-catalyzed reaction kinetics of several dye-labeled ribozyme variants for which precise optical activity determination (UV/vis, fluorescence) failed due to interference of the attached dyes. The reaction kinetics were analyzed by thin-layer chromatographic separation of the 32P-labeled reaction components and densitometric analysis of the substrate and product radioactivities, thereby allowing iterative optimization of the dye positions for future single-molecule studies. The phosphorylation strategy with trichloroacetonitrile may be applicable for labeling numerous other compounds that contain alcoholic hydroxyl groups. PMID:21731729

Agarose and Polyacrylamide Gel Electrophoresis Methods for Molecular Mass Analysis of 5–500 kDa Hyaluronan

PubMed Central

Bhilocha, Shardul; Amin, Ripal; Pandya, Monika; Yuan, Han; Tank, Mihir; LoBello, Jaclyn; Shytuhina, Anastasia; Wang, Wenlan; Wisniewski, Hans-Georg; de la Motte, Carol; Cowman, Mary K.

2011-01-01

Agarose and polyacrylamide gel electrophoresis systems for the molecular mass-dependent separation of hyaluronan (HA) in the size range of approximately 5–500 kDa have been investigated. For agarose-based systems, the suitability of different agarose types, agarose concentrations, and buffers systems were determined. Using chemoenzymatically synthesized HA standards of low polydispersity, the molecular mass range was determined for each gel composition, over which the relationship between HA mobility and logarithm of the molecular mass was linear. Excellent linear calibration was obtained for HA molecular mass as low as approximately 9 kDa in agarose gels. For higher resolution separation, and for extension to molecular masses as low as approximately 5 kDa, gradient polyacrylamide gels were superior. Densitometric scanning of stained gels allowed analysis of the range of molecular masses present in a sample, and calculation of weight-average and number-average values. The methods were validated for polydisperse HA samples with viscosity-average molecular masses of 112, 59, 37, and 22 kDa, at sample loads of 0.5 µg (for polyacrylamide) to 2.5 µg (for agarose). Use of the methods for electrophoretic mobility shift assays was demonstrated for binding of the HA-binding region of aggrecan (recombinant human aggrecan G1-IGD-G2 domains) to a 150 kDa HA standard. PMID:21684248

Stress degradation studies and development of stability-indicating TLC-densitometry method for determination of prednisolone acetate and chloramphenicol in their individual and combined pharmaceutical formulations

PubMed Central

2012-01-01

A rapid and reproducible stability indicating TLC method was developed for the determination of prednisolone acetate and chloramphenicol in presence of their degraded products. Uniform degradation conditions were maintained by refluxing sixteen reaction mixtures for two hours at 80°C using parallel synthesizer including acidic, alkaline and neutral hydrolysis, oxidation and wet heating degradation. Oxidation at room temperature, photochemical and dry heating degradation studies were also carried out. Separation was done on TLC glass plates, pre-coated with silica gel 60F-254 using chloroform: methanol (14:1 v/v). Spots at Rf 0.21 ± 0.02 and Rf 0.41 ± 0.03 were recognized as chloramphenicol and prednisolone acetate, respectively. Quantitative analysis was done through densitometric measurements at multiwavelength (243 nm, λmax of prednisolone acetate and 278 nm, λmax of chloramphenicol), simultaneously. The developed method was optimized and validated as per ICH guidelines. Method was found linear over the concentration range of 200-6000 ng/spot with the correlation coefficient (r2 ± S.D.) of 0.9976 ± 3.5 and 0.9920 ± 2.5 for prednisolone acetate and chloramphenicol, respectively. The developed TLC method can be applied for routine analysis of prednisolone acetate and chloramphenicol in presence of their degraded products in their individual and combined pharmaceutical formulations. PMID:22264235

Agarose-gel electrophoresis for the quality assurance and purity of heparin formulations.

PubMed

Volpi, Nicola; Buzzega, Dania

2012-01-01

The adulteration of raw heparin (Hep) with a synthetic oversulfated chondroitin sulfate (OSCS) not found in nature produced in 2007-2008 a global crisis giving rise to the development of additional, new and specific methods for its quality assurance and purity. In this study, a simple and sensitive agarose-gel electrophoresis method has been developed for the visualization of OSCS in Hep samples along with other natural glycosaminoglycans possibly present as "process-related impurities", in particular dermatan sulfate (DS) and chondroitin sulfate (CS). Agarose-gel electrophoresis under non-conventional conditions is able to separate OSCS from Hep with its two components, the slow-moving and fast-moving species, DS and CS by performing separation for 15 h (overnight) and under high voltage (100 mA, ∼200 V). Densitometric scanning enabled us to calculate a limit of detection of ∼0.5 μg OSCS with a linear behaviour from 0.1 to 5 μg, comparable to CS/DS. Contaminated samples from Hep manufacturers were analyzed and quantitative data were found comparable to previous studies. Due to its capacity to process many samples in a single run and to the equipment commonly available in laboratories, this analytical method would be suitable for the identification and quantification of contamination by other polysaccharides, in particular OSCS and DS, within Hep preparations and formulations. Copyright © 2012 Elsevier B.V. All rights reserved.

CNS development under altered gravity: cerebellar glial and neuronal protein expression in rat neonates exposed to hypergravity

NASA Astrophysics Data System (ADS)

Nguon, K.; Li, G.-H.; Sajdel-Sulkowska, E. M.

2004-01-01

The future of space exploration depends on a solid understanding of the developmental process under microgravity, specifically in relation to the central nervous system (CNS). We have previously employed a hypergravity paradigm to assess the impact of altered gravity on the developing rat cerebellum [Exp. Biol. Med. 226 (2000) 790]. The present study addresses the molecular mechanisms involved in the cerebellar response to hypergravity. Specifically, the study focuses on the expression of selected glial and neuronal cerebellar proteins in rat neonates exposed to hypergravity (1.5 G) from embryonic day (E)11 to postnatal day (P)6 or P9 (the time of maximal cerebellar changes) comparing them against their expression in rat neonates developing under normal gravity. Proteins were analyzed by quantitative Western blots of cerebellar homogenates; RNA analysis was performed in the same samples using quantitative PCR. Densitometric analysis of Western blots suggested a reduction in glial (glial acidic protein, GFAP) and neuronal (neuronal cell adhesion moiecule, NCAM-L1, synaptophysin) proteins, but the changes in individual cerebellar proteins in hypergravity-exposed neonates appeared both age- and gender-specific. RNA analysis suggested a reduction in GFAP and synaptophysin mRNAs on P6. These data suggest that exposure to hypergravity may interfere with the expression of selected cerebellar proteins. These changes in protein expression may be involved in mediating the effect of hypergravity on the developing rat cerebellum.

QUANTIFICATION OF GLYCYRRHIZIN BIOMARKER IN GLYCYRRHIZA GLABRA RHIZOME AND BABY HERBAL FORMULATIONS BY VALIDATED RP-HPTLC METHODS

PubMed Central

Alam, Prawez; Foudah, Ahmed I.; Zaatout, Hala H.; T, Kamal Y; Abdel-Kader, Maged S.

2017-01-01

Background: A simple and sensitive thin-layer chromatographic method has been established for quantification of glycyrrhizin in Glycyrrhiza glabra rhizome and baby herbal formulations by validated Reverse Phase HPTLC method. Materials and Methods: RP-HPTLC Method was carried out using glass coated with RP-18 silica gel 60 F254S HPTLC plates using methanol-water (7: 3 v/v) as mobile phase. Results: The developed plate was scanned and quantified densitometrically at 256 nm. Glycyrrhizin peaks from Glycyrrhiza glabra rhizome and baby herbal formulations were identified by comparing their single spot at Rf = 0.63 ± 0.01. Linear regression analysis revealed a good linear relationship between peak area and amount of glycyrrhizin in the range of 2000-7000 ng/band. Conclusion: The method was validated, in accordance with ICH guidelines for precision, accuracy, and robustness. The proposed method will be useful to enumerate the therapeutic dose of glycyrrhizin in herbal formulations as well as in bulk drug. PMID:28573236

Determination of cortisol in human plasma by thin-layer chromatography and fluorescence derivatization with isonicotinic acid hydrazide.

PubMed

Fenske, Martin

2008-01-01

The present work describes a specific and rapid determination of cortisol in human plasma. The method includes liquid-liquid extraction of plasma samples, thin-layer chromatography (TLC) of ethanolic extracts on aluminium foil-backed silica gel 60 TLC plates, derivatization of cortisol with isonicotinic acid hydrazide, and densitometric measurement of the fluorescence intensity of cortisol hydrazone. The fluorescence was linearly related to cortisol amounts; the correlation coefficients of standard curve plots were r>0.99. The coefficient of variation ranged between 2.8-7.9% (20 ng, within-assay/between assay variation) and 1.6-6.8% (80 ng, within-assay/between assay variation). The recovery of cortisol from plasma spiked with 21-deoxycortisol was 85%+/-4%. Cortisol concentration in the plasma was 66+/-32 ng/mL (mean+/-standard deviation, n=24). The advantage of this method is its simplicity to separate cortisol from other steroids by TLC, its specificity (formation of cortisol hydrazone), and the rapid quantitation of cortisol by densitometry.

Quantification through TLC-densitometric analysis, repellency and anticholinesterase activity of the homemade extract of Indian cloves.

PubMed

Affonso, Raphael S; Lima, Josélia A; Lessa, Bruno M; Caetano, João V O; Obara, Marcos T; Nóbrega, Andréa B; Nepovimova, Eugenie; Musilek, Kamil; Kuca, Kamil; Slana, Gláucia B C A; França, Tanos C C

2018-02-01

The rise of the mosquitoes-transmitted diseases, like dengue, zika and chikungunya in Brazil in the last years has increased concerns on protection against mosquitoes bites. However, the prohibitive prices of the commercially available repellents for the majority of the Brazilian population has provoked a search for cheaper solutions, like the use of the homemade ethanolic extract of Indian clove (Syzygium aromaticum L.) as repellent, which has been reported as quite efficient by the local press. In order to verify this, we performed here the quantification of the main components of this extract through high-performance thin-layer chromatography (HPTLC)-densitometry and evaluated its efficiency as a repellent and its acetylcholinesterase (AChE) inhibition capacity. Our results have proved HPTLC-densitometry as an efficient and appropriate method for this quantification and confirmed the repellency activity, as well as its capacity of AChE inhibition. Copyright © 2017 John Wiley & Sons, Ltd.

Simultaneous quantification of withanolides in Withania somnifera by a validated high-performance thin-layer chromatographic method.

PubMed

Srivastava, Pooja; Tiwari, Neerja; Yadav, Akhilesh K; Kumar, Vijendra; Shanker, Karuna; Verma, Ram K; Gupta, Madan M; Gupta, Anil K; Khanuja, Suman P S

2008-01-01

This paper describes a sensitive, selective, specific, robust, and validated densitometric high-performance thin-layer chromatographic (HPTLC) method for the simultaneous determination of 3 key withanolides, namely, withaferin-A, 12-deoxywithastramonolide, and withanolide-A, in Ashwagandha (Withania somnifera) plant samples. The separation was performed on aluminum-backed silica gel 60F254 HPTLC plates using dichloromethane-methanol-acetone-diethyl ether (15 + 1 + 1 + 1, v/v/v/v) as the mobile phase. The withanolides were quantified by densitometry in the reflection/absorption mode at 230 nm. Precise and accurate quantification could be performed in the linear working concentration range of 66-330 ng/band with good correlation (r2 = 0.997, 0.999, and 0.996, respectively). The method was validated for recovery, precision, accuracy, robustness, limit of detection, limit of quantitation, and specificity according to International Conference on Harmonization guidelines. Specificity of quantification was confirmed using retention factor (Rf) values, UV-Vis spectral correlation, and electrospray ionization mass spectra of marker compounds in sample tracks.

Simultaneous HPTLC Determination of Rabeprazole and Itopride Hydrochloride From Their Combined Dosage Form.

PubMed

Suganthi, A; John, Sofiya; Ravi, T K

2008-01-01

A simple, precise, sensitive, rapid and reproducible HPTLC method for the simultaneous estimation of the rabeprazole and itopride hydrochloride in tablets was developed and validated. This method involves separation of the components by TLC on precoated silica gel G60F254 plate with solvent system of n-butanol, toluene and ammonia (8.5:0.5:1 v/v/v) and detection was carried out densitometrically using a UV detector at 288 nm in absorbance mode. This system was found to give compact spots for rabeprazole (Rf value of 0.23 0.02) and for itopride hydrochloride (Rf value of 0.75+/-0.02). Linearity was found to be in the range of 40-200 ng/spot and 300-1500 ng/spot for rabeprazole and itopride hydrochloride. The limit of detection and limit of quantification for rabeprazole were 10 and 20 ng/spot and for itopride hydrochloride were 50 and 100 ng/spot, respectively. The method was found to be beneficial for the routine analysis of combined dosage form.

RP-HPTLC densitometric determination and validation of vanillin and related phenolic compounds in accelerated solvent extract of Vanilla planifolia*.

PubMed

Sharma, Upendra Kumar; Sharma, Nandini; Gupta, Ajai Prakash; Kumar, Vinod; Sinha, Arun Kumar

2007-12-01

A simple, fast and sensitive RP-HPTLC method is developed for simultaneous quantitative determination of vanillin and related phenolic compounds in ethanolic extracts of Vanilla planifolia pods. In addition to this, the applicability of accelerated solvent extraction (ASE) as an alternative to microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE) and Soxhlet extraction was also explored for the rapid extraction of phenolic compounds in vanilla pods. Good separation was achieved on aluminium plates precoated with silica gel RP-18 F(254S) in the mobile phase of methanol/water/isopropanol/acetic acid (30:65:2:3, by volume). The method showed good linearity, high precision and good recovery of compounds of interest. ASE showed good extraction efficiency in less time as compared to other techniques for all the phenolic compounds. The present method would be useful for analytical research and for routine analysis of vanilla extracts for their quality control.

Qualitative and quantitative analysis of seven oligosaccharides in Morinda officinalis using double-development HPTLC and scanning densitometry.

PubMed

Zhou, Bin; Chang, Jun; Wang, Ping; Li, Jie; Cheng, Dan; Zheng, Peng-Wu

2014-01-01

The quality of Morindaofficinalis, which has been used as a Yang-tonic agent for a long time in China, can be evaluated. A double-development high performance thin layer chromatography (HPTLC) method has been established to simultaneously analyze quality and quantity of seven inulin-type oligosaccharides (DP=3-9) in Morindaofficinalis. The chromatography was performed on a silica gel 60 plate with the 7:5:2:1 proportion (v/v) of n-butanol-isopropanol-water-acetic acid for the first and second developments, respectively. The bands were visualized by the reaction with aniline-diphenylamine-phosphoric acid solution and analyzed by densitometric TLC at 540 nm. Quantification of seven oligosaccharides was achieved by densitometry at 540 nm. The investigated standard sugar had good linearity (R2>0.99) within test ranges. The amounts of seven oligosaccharides were calculated by the relative correction factor (RCF). Therefore, the developed TLC method could be used for quality control of Morindaofficinalis.

Bone-marrow densitometry: Assessment of marrow space of human vertebrae by single energy high resolution-quantitative computed tomography

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

Peña, Jaime A.; Damm, Timo; Bastgen, Jan

Purpose: Accurate noninvasive assessment of vertebral bone marrow fat fraction is important for diagnostic assessment of a variety of disorders and therapies known to affect marrow composition. Moreover, it provides a means to correct fat-induced bias of single energy quantitative computed tomography (QCT) based bone mineral density (BMD) measurements. The authors developed new segmentation and calibration methods to obtain quantitative surrogate measures of marrow-fat density in the axial skeleton. Methods: The authors developed and tested two high resolution-QCT (HR-QCT) based methods which permit segmentation of bone voids in between trabeculae hypothesizing that they are representative of bone marrow space. Themore » methods permit calculation of marrow content in units of mineral equivalent marrow density (MeMD). The first method is based on global thresholding and peeling (GTP) to define a volume of interest away from the transition between trabecular bone and marrow. The second method, morphological filtering (MF), uses spherical elements of different radii (0.1–1.2 mm) and automatically places them in between trabeculae to identify regions with large trabecular interspace, the bone-void space. To determine their performance, data were compared ex vivo to high-resolution peripheral CT (HR-pQCT) images as the gold-standard. The performance of the methods was tested on a set of excised human vertebrae with intact bone marrow tissue representative of an elderly population with low BMD. Results: 86% (GTP) and 87% (MF) of the voxels identified as true marrow space on HR-pQCT images were correctly identified on HR-QCT images and thus these volumes of interest can be considered to be representative of true marrow space. Within this volume, MeMD was estimated with residual errors of 4.8 mg/cm{sup 3} corresponding to accuracy errors in fat fraction on the order of 5% both for GTP and MF methods. Conclusions: The GTP and MF methods on HR-QCT images permit noninvasive localization and densitometric assessment of marrow fat with residual accuracy errors sufficient to study disorders and therapies known to affect bone marrow composition. Additionally, the methods can be used to correct BMD for fat induced bias. Application and testing in vivo and in longitudinal studies are warranted to determine the clinical performance and value of these methods.« less

Development and Validation of a HPTLC Method for Simultaneous Estimation of L-Glutamic Acid and γ-Aminobutyric Acid in Mice Brain

PubMed Central

Sancheti, J. S.; Shaikh, M. F.; Khatwani, P. F.; Kulkarni, Savita R.; Sathaye, Sadhana

2013-01-01

A new robust, simple and economic high performance thin layer chromatographic method was developed for simultaneous estimation of L-glutamic acid and γ-amino butyric acid in brain homogenate. The high performance thin layer chromatographic separation of these amino acid was achieved using n-butanol:glacial acetic acid:water (22:3:5 v/v/v) as mobile phase and ninhydrin as a derivatising agent. Quantitation of the method was achieved by densitometric method at 550 nm over the concentration range of 10-100 ng/spot. This method showed good separation of amino acids in the brain homogenate with Rf value of L-glutamic acid and γ-amino butyric acid as 21.67±0.58 and 33.67±0.58, respectively. The limit of detection and limit of quantification for L-glutamic acid was found to be 10 and 20 ng and for γ-amino butyric acid it was 4 and 10 ng, respectively. The method was also validated in terms of accuracy, precision and repeatability. The developed method was found to be precise and accurate with good reproducibility and shows promising applicability for studying pathological status of disease and therapeutic significance of drug treatment. PMID:24591747

Development and Validation of a HPTLC Method for Simultaneous Estimation of L-Glutamic Acid and γ-Aminobutyric Acid in Mice Brain.

PubMed

Sancheti, J S; Shaikh, M F; Khatwani, P F; Kulkarni, Savita R; Sathaye, Sadhana

2013-11-01

A new robust, simple and economic high performance thin layer chromatographic method was developed for simultaneous estimation of L-glutamic acid and γ-amino butyric acid in brain homogenate. The high performance thin layer chromatographic separation of these amino acid was achieved using n-butanol:glacial acetic acid:water (22:3:5 v/v/v) as mobile phase and ninhydrin as a derivatising agent. Quantitation of the method was achieved by densitometric method at 550 nm over the concentration range of 10-100 ng/spot. This method showed good separation of amino acids in the brain homogenate with Rf value of L-glutamic acid and γ-amino butyric acid as 21.67±0.58 and 33.67±0.58, respectively. The limit of detection and limit of quantification for L-glutamic acid was found to be 10 and 20 ng and for γ-amino butyric acid it was 4 and 10 ng, respectively. The method was also validated in terms of accuracy, precision and repeatability. The developed method was found to be precise and accurate with good reproducibility and shows promising applicability for studying pathological status of disease and therapeutic significance of drug treatment.

Stability-Indicating HPTLC Method for Simultaneous Estimation of Flurbiprofen and Chloramphenicol in Ophthalmic Solution.

PubMed

Sadakwala, Vaishnavi M; Chauhan, Renu S; Shah, Shailesh A; Shah, Dinesh R

2016-01-01

A specific, accurate and reproducible stability-indicating high performance thin layer chromatography (HPTLC) method was developed for the estimation of flurbiprofen and chloramphenicol in the presence of their degradation products. Degradation studies of both the drugs were carried out in acidic, alkaline, neutral, oxidative, photolytic and thermal stress conditions. Separation was performed on thin layer chromatography plate precoated with silica gel 60 F254 using ethyl acetate : n-hexane : methanol : tri-ethyl amine (5 : 4 : 2 : 0.5, v/v/v/v). Spots at retention factor 0.29 and 0.62 were recognized as flurbiprofen and chloramphenicol, respectively, and were quantified through densitometric measurements at wavelength 267 nm. Method was found to be linear over the concentration range 12-60 ng/spot with correlation coefficient of 0.9997 for flurbiprofen and 200-1,000 ng/spot with correlation coefficient of 0.9977 for chloramphenicol. The proposed method was applied to the estimation of flurbiprofen and chloramphenicol in commercial ophthalmic formulation. The developed HPTLC method can be applied for routine analysis of flurbiprofen and chloramphenicol in the presence of their degradation products in their individual as well as combined pharmaceutical formulations. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Purification of two high molecular weight proteases from rabbit reticulocyte lysate

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

Hough, R.; Pratt, G.; Rechsteiner, M.

1987-05-01

The authors have purified two large proteases from rabbit reticulocyte lysate. The enzymes are so similar in their chromatographic behavior that each is the only significant contaminant of the other during the final stages of purification. At pH 7.8, both hydrolyze /sup 125/I-..cap alpha..-casein and 4-methylcoumaryl-7-amide (MCA) derivatives with tyrosine, phenylalanine or arginine at the P/sub 1/ position. The larger, ATP-dependent enzyme degrades ubiquitin-lysozyme conjugates, but it does not degrade unmodified lysozyme. Hydrolysis of Suc-Leu-Leu-Val-Tyr-MCA by this enzyme is also stimulated two-fold in the presence of ATP. The protease has a molecular weight of 950,000 based on sedimentation, gel filtrationmore » and non-denaturing PAGE. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the protease is composed of a number of subunits with molecular masses between 32 and 110 kDa. Densitometric analysis showed equivalent amounts of the two larger chains, and the presence of one copy of each in the native enzyme would be consistent with an M/sub r/ of 950,000. The smaller protease has a molecular weight of 700,000 and is composed of 8 to 10 subunits ranging from 21,000 to 32,000. It cleaves ubiquitin-lysozyme conjugates only slightly, and hydrolysis of conjugates or fluorogenic peptide substrates is not stimulated by ATP. This protease appears similar, if not identical, to the multicatalytic protease complex first purified by Wilk and Orlowski.« less

Thalassemic osteopathy: a new marker of bone deposition.

PubMed

Baldini, M; Forti, S; Orsatti, A; Marcon, A; Ulivieri, F M; Airaghi, L; Zanaboni, L; Cappellini, M D

2014-01-01

Osteopathy represents a prominent cause of morbidity in patients with beta-thalassemia major (TM) and manifests as osteopenia/osteoporosis. Biochemical turnover markers (BTMs) are considered a useful, non-invasive tool for the clinical follow-up of osteoporotic patients; they can provide a dynamic view of the remodeling process and give information on the metabolic activity of bone tissue as well as on the pathogenesis of bone loss. The amino-terminal pro-peptide of type I procollagen (P1NP) is a recently introduced marker that is considered the most sensitive index of bone formation. Although demonstrated in several categories of patients with bone disease, there is little information on the clinical usefulness of this bone formation index in thalassemic patients. We evaluated the P1NP levels of 53 adult patients with b-thalassemia major (21 males and 32 females, mean age 34.5 ± 5.7, range 22-46 years) and associated osteopathy. We investigated the correlation between P1NP and bone condition as examined by dual X-ray photon absorptiometry and with BTMs expressing bone resorption and bone mineralization (carboxyterminal collagen cross-linked (CTX) terminal regions of type I collagen and osteocalcin, respectively). P1NP serum levels were correlated with CTX levels (r=0.545, p<0.001); the results were unchanged when males and females, as well as osteoporotic and osteopenic subgroups, were considered separately. No correlation was demonstrated neither between OC and CTX (r=0.17, p=ns), nor between P1NP and OC levels (r=0.11, p=ns). No correlation was demonstrated among the P1NP/CTX ratio and age, OC or densitometric values and no difference was found in the same ratio between osteopenic (0.19 ± 0.16) and osteoporotic (0.15 ± 0.14) patients. Similar results were obtained for the OC/CTX ratio, as it was not correlated with age, P1NP or densitometric values. This is the first report of circulating P1NP in patients with TM-associated osteoporosis. P1NP and CTX assays show good precision and low analytical CV, and, compared to other markers, they can acceptably reflect bone metabolic processes and promptly respond to antiosteoporotic treatments. We trust that this sensitive marker can be useful in the assessment of treatment efficacy and can overcome the pitfalls due to wide variability in the normal values of most BTMs that create difficulty in pinpointing the individual patient's response. © 2013.

[Total body composition in adult patients with growth hormone deficiency before and after its administration].

PubMed

Weiss, V; Krsek, M; Marek, J; Stĕpán, J; Malík, J

2003-08-01

Effect of growth hormone (GH) on the growth and development of children is generally known. Effects of GH in adults are favorable, though. The aim of the work was to verify effects of GH administration on body composition in adult patients with GH deficit (GHD). The authors examined 15 adult patients with GHD originated in 13 of them in adulthood and in two of them in childhood. Their mean age was 43.9 +/- 11.3 years, the mean body mass was 80.0 +/- 15.2 kg. The GH deficit was verified by the stimulation insulin tolerance test. For the period of 12 months, they were subcutaneously administered recombinant human GH in a substitution dose of 0.5 to 1.5 IU/m2 body surface/day. A stable substitution of the hormone was applied for the period of at least six months in all these patients provided any deficit of other hormones had not been demonstrated. The examination by whole-body dosimeter Lunar DPX-L was made in the patients before the GH treatment began and after 12 months of therapy. It enabled to determine the amount of lean body mass (LBM) and fatty mass. After 12 months of GH treatment the mean level of insulin-like growth factor (IGF-I) was increased (P = 0.002). A statistically significant increase of total LBM (48.6 +/- 9.8 vs. 50.8 +/- 9.9 kg, P = 0.004) developed, the fatty mass did not change. Nine of these 15 patients were further followed and the administration of GH proceeded for six months. The densitometric examination was repeated, but no change of LBM was observed. The administration of GH was halted and after the period of 12 months the whole-body densitometric examination was done. The increase of LBM lasted. The amount of fat mass did not change, a decrease of fatty mass was observed after the GH administration ended. After 12 months of GH treatment there was also an increase of maximal output reached on bicycle ergometer (157.3 +/- 34.2 vs. 197.5 +/- 68.1 W, P = 0.006). A positive correlation between LBM and maximal output reached on bicycle ergometer before GH administration (r = 0.58, P = 0.02) was observed. A favorable effect of the substitution dose of GS administered to adult patients with GHD on the increase of LBM and physical output was confirmed.

[The incidence of osteopenia and osteoporosis in patients with cementless total hip arthroplasty].

PubMed

Lacko, M; Schreierová, D; Čellár, R; Vaško, G

2015-01-01

The aim of the study was to evaluate the incidence of osteopenia and osteoporosis in the patients elected to cementless total hip replacement. The group evaluated comprised 100 patients with primary or secondary forms of coxarthrosis who underwent cementless total hip arthroplasty (THA). The results of densitometric examination of the lumbar spine and proximal femur were analysed. Based on the lowest T-score value, the patients were divided into three groups, i.e., fist, normal bone density; second, osteopenia; third, osteoporosis. Clinical examination included patient medical history, Harris hip scores and visual analogue scale assessment for pain intensity; the Kellgren-Lawrence classification was used to measure the grade of hip osteoarthritis; blood tests were made to assess the levels of total calcium, ionised calcium, phosphorus, vitamin D and the markers of bone resorption and formation. Osteoporosis was found in 32 and osteopenia in 21 patients; 47 patients had normal bone density. Osteoporosis was detected in the lumbar spine of 21 patients, in the proximal femur of nine patients and at both sites in two patients. In 13 patients this diagnosis was made for the fist time. The patients with osteoporosis had a significantly lower body mass index. There were no differences in the other characteristics, i.e., age, functional, radiological and laboratory findings, among the groups; all three showed the mean vitamin D concentration below the lowest level of its physiological range. Insufficient vitamin D levels were found in 54 women and 18 men. Of all patients, only 13 women and four men took vitamin D supplements. DISCUSSION Osteoarthritis and osteoporosis are the most frequent complex musculoskeletal diseases. Several studies have suggested that these disorders are mutually exclusive. This assumption has been based on the absence of radiographic evidence of osteoarthritis seen in many elderly patients with femoral neck fractures. Our relatively frequent findings of osteopenia and osteoporosis in this study show that patients with osteoarthritis may not be protected from the development of generalised osteoporosis. Our results are compared with the relevant literature data and potential effects of osteoporosis and vitamin D deficiency on total hip arthroplasty are discussed. Hip osteoarthritis does not reduce the risk of generalised osteoporosis. We found a relative high incidence of osteopenia and osteoporosis as well as vitamin D deficiency in patient with advanced form of hip osteoarthritis.

Stability-indicating chromatographic methods for the determination of some oxicams.

PubMed

Taha, Elham Anwer; Salama, Nahla Nour; Abdel Fattah, Laila el-Said

2004-01-01

Two sensitive and selective methods were developed for the determination of some oxicams, namely, lornoxicam (LOX), tenoxicam (TEX), and meloxicam (MEX), in the presence of their alkaline degradation products. The first method is based on the thin-layer chromatographic separation of the 3 drugs from their alkaline degradation products, followed by densitometric measurement of the intact drug spots for LOX, TEX, and MEX at 380, 370, and 364 nm, respectively. The developing systems used for separation are ethyl acetate-methanol-26% ammonia (17 + 3 + 0.35, v/v/v) for LOX and TEX and chloroform-n-hexane-96.0% acetic acid (18 + 1 + 1, v/v/v) for MEX. The linear ranges were 0.25-6.0 microg/spot for LOX and TEX and 0.5-10 microg/spot for MEX, with mean recoveries of 99.80 +/- 1.32, 100.57 +/- 1.34, and 100.71 +/- 1.57%, respectively. The second method is based on the liquid chromatographic separation of the 3 drugs from their alkaline degradation products on a reversed-phase C18 column, using mobile phases of methanol-acetonitrile-acetate buffer, pH 4.6 (4.5 + 0.5 + 5.0, v/v/v) for LOX and MEX and methanol-acetonitrile-acetate buffer, pH 4.6 (1.9 + 0.1 + 3.0, v/v/v) for TEX at ambient temperature. Quantification is achieved by UV detection at 280 nm, based on peak area. The linear ranges were 0.5-20 microg/mL for LOX and TEX and 1.25-50 microg/mL for MEX, with mean recoveries of 99.81 +/- 1.01, 98.90 +/- 1.61, and 100.86 +/- 1.55%, respectively. The methods were validated according to guidelines of the International Conference on Harmonization. The developed methods were successfully applied to the determination of LOX, TEX, and MEX in bulk powder, laboratory-prepared mixtures containing different percentages of degradation products, and pharmaceutical dosage forms.

Proteolytic roles of matrix metalloproteinase (MMP)-13 during progression of chronic periodontitis: initial evidence for MMP-13/MMP-9 activation cascade.

PubMed

Hernández Ríos, Marcela; Sorsa, Timo; Obregón, Fabián; Tervahartiala, Taina; Valenzuela, María Antonieta; Pozo, Patricia; Dutzan, Nicolás; Lesaffre, Emmanuel; Molas, Marek; Gamonal, Jorge

2009-12-01

Matrix metalloproteinases (MMP)-13 can initiate bone resorption and activate proMMP-9 in vitro, and both these MMPs have been widely implicated in tissue destruction associated with chronic periodontitis. We studied whether MMP-13 activity and TIMP-1 levels in gingival crevicular fluid (GCF) associated with progression of chronic periodontitis assessed clinically and by measuring carboxy-terminal telopeptide of collagen I (ICTP) levels. We additionally addressed whether MMP-13 could potentiate gelatinase activation in diseased gingival tissue. In this prospective study, GCF samples from subjects undergoing clinical progression of chronic periodontitis and healthy controls were screened for ICTP levels, MMP-13 activity and TIMP-1. Diseased gingival explants were cultured, treated or not with MMP-13 with or without adding CL-82198, a synthetic MMP-13 selective inhibitor, and assayed by gelatin zymography and densitometric analysis. Active sites demonstrated increased ICTP levels and MMP-13 activity (p<0.05) in progression subjects. The MMP-9 activation rate was elevated in MMP-13-treated explants (p<0.05) and MMP-13 inhibitor prevented MMP-9 activation. MMP-13 could be implicated in the degradation of soft and hard supporting tissues and proMMP-9 activation during progression of chronic periodontitis. MMP-13 and -9 can potentially form an activation cascade overcoming the protective TIMP-1 shield, which may become useful for diagnostic aims and a target for drug development.

CNS development under altered gravity: cerebellar glial and neuronal protein expression in rat neonates exposed to hypergravity

NASA Technical Reports Server (NTRS)

Nguon, K.; Li, G-H; Sajdel-Sulkowska, E. M.

2004-01-01

The future of space exploration depends on a solid understanding of the developmental process under microgravity, specifically in relation to the central nervous system (CNS). We have previously employed a hypergravity paradigm to assess the impact of altered gravity on the developing rat cerebellum. The present study addresses the molecular mechanisms involved in the cerebellar response to hypergravity. Specifically, the study focuses on the expression of selected glial and neuronal cerebellar proteins in rat neonates exposed to hypergravity (1.5 G) from embryonic day (E)11 to postnatal day (P)6 or P9 (the time of maximal cerebellar changes) comparing them against their expression in rat neonates developing under normal gravity. Proteins were analyzed by quantitative Western blots of cerebellar homogenates; RNA analysis was performed in the same samples using quantitative PCR. Densitometric analysis of Western blots suggested a reduction in glial (glial acidic protein, GFAP) and neuronal (neuronal cell adhesion molecule, NCAM-L1, synaptophysin) proteins, but the changes in individual cerebellar proteins in hypergravity-exposed neonates appeared both age- and gender-specific. RNA analysis suggested a reduction in GFAP and synaptophysin mRNAs on P6. These data suggest that exposure to hypergravity may interfere with the expression of selected cerebellar proteins. These changes in protein expression may be involved in mediating the effect of hypergravity on the developing rat cerebellum. c2003 COSPAR. Published by Elsevier Ltd. All rights reserved.

HPTLC method for the simultaneous determination of four indole alkaloids in Rauwolfia tetraphylla: a study of organic/green solvent and continuous/pulse sonication.

PubMed

Gupta, Shikha; Shanker, Karuna; Srivastava, Santosh K

2012-07-01

A new validated high-performance thin-layer chromatographic (HPTLC) method has been developed for the simultaneous quantitation of four antipsychotic indole alkaloids (IAs), reserpiline (RP, 1), α-yohimbine (YH, 2), isoreserpiline (IRP, 3) and 10-methoxy tetrahydroalstonine (MTHA, 4) as markers in the leaves of Rauwolfia tetraphylla. Extraction efficiency of the targeted IAs from the leaf matrix with organic and ecofriendly (green) solvents using percolation, ultrasonication and microwave techniques were studied. Non-ionic surfactants, viz. Triton X-100, Triton X-114 and Genapol X-80 were used for extraction and no back-extraction or liquid chromatographic steps were used to remove the targeted IAs from the surfactant-rich extractant phase. The optimized cloud point extraction was found a potentially useful methodology for the preconcentration of the targeted IAs. The separation was achieved on silica gel 60F(254) HPTLC plates using hexane-ethylacetate-methanol (5:4:1, v/v/v) as mobile phase. The quantitation of IAs (1-4) was carried out using the densitometric reflection/absorption mode at 520 nm after post chromatographic derivatization using Dragendorff's reagent. The method was validated for peak purity, precision, accuracy, robustness, limit of detection (LOD) and quantitation (LOQ). Method specificity was confirmed using retention factor (R(f)) and visible spectral (post chromatographic scan) correlation of marker compounds in the samples and standard tracks. Copyright © 2012 Elsevier B.V. All rights reserved.

Computerized tomographic quantification of chronic obstructive pulmonary disease as the principal determinant of frontal P vector.

PubMed

Chhabra, Lovely; Sareen, Pooja; Gandagule, Amit; Spodick, David

2012-04-01

Verticalization of the P-wave axis is characteristic of chronic obstructive pulmonary disease (COPD). We studied the correlation of P-wave axis and computerized tomographically quantified emphysema in patients with COPD/emphysema. Individual correlation of P-wave axis with different structural types of emphysema was also studied. High-resolution computerized tomographic scans of 23 patients >45 years old with known COPD were reviewed to assess the type and extent of emphysema using computerized tomographic densitometric parameters. Electrocardiograms were then independently reviewed and the P-wave axis was calculated in customary fashion. Degree of the P vector (DOPV) and radiographic percent emphysematous area (RPEA) were compared for statistical correlation. The P vector and RPEA were also directly compared to the forced expiratory volume at 1 second. RPEA and the P vector had a significant positive correlation in all patients (r = +0.77, p <0.0001) but correlation was very strong in patients with predominant lower lobe emphysema (r = +0.89, p <0.001). Forced expiratory volume at 1 second and the P vector had almost a linear inverse correlation in predominantly lower lobe emphysema (r = -0.92, p <0.001). DOPV positively correlated with radiographically quantified emphysema. DOPV and RPEA were strong predictors of qualitative lung function in patients with predominantly lower lobe emphysema. In conclusion, a combination of high DOPV and predominantly lower lobe emphysema indicates severe obstructive lung dysfunction in patients with COPD. Copyright © 2012 Elsevier Inc. All rights reserved.

Autoradiographic disposition of (1-methyl-/sup 14/C)- and (2-/sup 14/C)caffeine in mice

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

Lachance, M.P.; Marlowe, C.; Waddell, W.J.

1983-11-01

Male, C57B1/6J mice received either (1-methyl-14C)caffeine or (2-14C)caffeine via the tail vein at a dose of 0.7 or 11 mg/kg, respectively. At 0.1, 0.33, 1, 3, 9, and 24 hr after treatment, the mice were anesthetized with ether and frozen by immersion in dry ice/hexane. The mice were processed for whole-body autoradiography by the Ullberg technique; this procedure does not allow thawing or contact with solvents. All autoradiographs revealed some retention of radioactivity at early time intervals in the lacrimal glands, seminal vesicle fluid, nasal and olfactory epithelium, and retinal melanocytes. The remaining portion of the animal was densitometrically uniformmore » except for the lower levels noted in the CNS and adipose tissues. Excretion of radioactivity by the liver and kidneys seems to be the major routes of elimination. Localization in the liver at late time intervals was confined principally to the centrilobular region. Late sites of retention, observed only after (1-methyl-14C)caffeine administration, included the pancreas, minor and major salivary glands, splenic red pulp, thymal cortex, bone marrow, and gastrointestinal epithelium. Sites of localization present in both studies included the olfactory epithelium, lacrimal glands, hair follicles, and retinal melanocytes. Further studies are needed to determine whether the localization at these various sites is due to metabolic degradation, active transport, or possibly a specific receptor interaction.« less

Management of natural resources through automatic cartographic inventory

NASA Technical Reports Server (NTRS)

Rey, P.; Gourinard, Y.; Cambou, F. (Principal Investigator)

1973-01-01

The author has identified the following significant results. Over those parts of the ARNICA test site where ERTS-1 data were available, the search for correspondences between images and ground truth acquired by the vegetation and geology maps was quite positive. The probability of recognition of soil use types can be estimated at: (1) 100% for water plans, rivers, canals, swamplands, and wetlands; (2) 80%-100% for the major types of forestry, farmland zones, moorlands and pasturelands, and urbanization; (3) 20%-50% for communication lines; (4) 60%-80% for forestry species and organization of agricultural areas; (5) 40%-60% for finer discrimination between forest types and more accurate identification of cultivations; (6) 60%-90% for major geological features. These percentages will be improved upon as soon as it is possible to use the repetitive imagery. An early use of automatic cartography using ERTS-1 imagery was made possible for pine forests in the Central Pyrenees, the densitometric signature of which were particularly significant. Important observations were made in related fields of water resources, snow survey, estuary dynamics, and meteorology.

7-Ketocholesterol and 7-hydroxycholesterol in pork meat and its gravy thermally treated without additives and in the presence of onion and garlic.

PubMed

Janoszka, Beata

2010-12-01

The effect of onion and garlic on the formation of two cholesterol oxidation products (COPs): 7-ketocholesterol and 7-hydroxycholesterol was evaluated by comparing their concentrations in meat and gravy samples obtained from three pork dishes prepared in the presence and absence of these flavourings. The concentration of these compounds in meat samples was between 82.4 and 1331.6 ng/g of cooked meat. Gravies contained lower amounts: from 18.3 to 45.6 ng/g of cooked meat. The addition of onion (30 g/100g of meat) caused a decrease in 7-ketocholesterol and 7-hydroxycholesterol concentrations in all of the investigated pork dishes by 9.5-79%, whilst the addition of 15 g of garlic to 100g of meat lowered the concentration by 17 to 88%. The greatest decrease was found in grilled minced chops. The quantitative assessment of 7-ketocholesterol and 7-hydroxycholesterol was carried out by thin-layer chromatography with densitometric detection. Copyright © 2010 The American Meat Science Association. Published by Elsevier Ltd.. All rights reserved.

Immunoturbidimetric assay for estimating free light chains of immunoglobulins in urine and serum.

PubMed Central

Tillyer, C R; Iqbal, J; Raymond, J; Gore, M; McIlwain, T J

1991-01-01

An immunoturbidimetric assay for the assessment of free kappa and lambda light chains of immunoglobulins was developed using a commercial polyclonal antiserum with reactivity towards epitopes on the light chains, which are not expressed when they are bound to heavy chains. The assay, on a centrifugal analyser, is simple and rapid. The limit of detection is 5 mg/l of free light chain, with an assay range of 5-120 mg/l, intrabatch precisions from 1.5-6.4%, and interbatch precisions from 6.5-8.9%. The assay was only slightly less sensitive than colloidal gold staining of cellulose acetate electrophoreses for the detection of Bence-Jones protein in urine. For the serial monitoring of response to chemotherapy in patients with myeloma, the assay correlated well with serum paraprotein estimates obtained by densitometric scanning of Ponceau stained cellulose acetate electrophoreses, but not with serum beta-2 microglobulin measurements, even after correction for the effects of creatinine. These assays may prove to be of use for the monitoring of tumour response in the treatment of Bence-Jones myeloma. PMID:1906071

Development and validation of a HPTLC method for simultaneous estimation of lornoxicam and thiocolchicoside in combined dosage form.

PubMed

Sahoo, Madhusmita; Syal, Pratima; Hable, Asawaree A; Raut, Rahul P; Choudhari, Vishnu P; Kuchekar, Bhanudas S

2011-07-01

To develop a simple, precise, rapid and accurate HPTLC method for the simultaneous estimation of Lornoxicam (LOR) and Thiocolchicoside (THIO) in bulk and pharmaceutical dosage forms. The separation of the active compounds from pharmaceutical dosage form was carried out using methanol:chloroform:water (9.6:0.2:0.2 v/v/v) as the mobile phase and no immiscibility issues were found. The densitometric scanning was carried out at 377 nm. The method was validated for linearity, accuracy, precision, LOD (Limit of Detection), LOQ (Limit of Quantification), robustness and specificity. The Rf values (±SD) were found to be 0.84 ± 0.05 for LOR and 0.58 ± 0.05 for THIO. Linearity was obtained in the range of 60-360 ng/band for LOR and 30-180 ng/band for THIO with correlation coefficients r(2) = 0.998 and 0.999, respectively. The percentage recovery for both the analytes was in the range of 98.7-101.2 %. The proposed method was optimized and validated as per the ICH guidelines.

Profiling of components and validated determination of iridoids in Gardenia Jasminoides Ellis fruit by a high-performance-thin-layer- chromatography/mass spectrometry approach.

PubMed

Coran, Silvia A; Mulas, Stefano; Vasconi, Alessio

2014-01-17

A novel method was set up with the aim to obtain a simultaneous cross comparative evaluation of different Gardenia Jasminoides Ellis fruits by the HPTLC fingerprint approach. The main components among the iridoid, hydroxycinnamic derivative and crocin classes were identified by TLC-MS ancillary techniques. The iridoids geniposide, gardenoside and genepin-1-β-d-gentiobioside were also quantitated by densitometric scanning at 240nm. LiChrospher HPTLC Silica gel 60 RP-18 W F254, 20cm×10cm plates with acetonitrile: formic acid 0.1% (40:60 v/v) as the mobile phase was used. The method was validated giving rise to a dependable and high throughput procedure well suited to routine applications. Iridoids were quantified in the range of 240-1140ng with RSD of repeatability and intermediate precision between 0.9-2.5% and accuracy with bias 1.6-2.6%. The method was tested on six commercial Gardenia Jasminoides fruit samples. Copyright © 2013 Elsevier B.V. All rights reserved.

Simultaneous HPTLC Determination of Rabeprazole and Itopride Hydrochloride From Their Combined Dosage Form

PubMed Central

Suganthi, A.; John, Sofiya; Ravi, T. K.

2008-01-01

A simple, precise, sensitive, rapid and reproducible HPTLC method for the simultaneous estimation of the rabeprazole and itopride hydrochloride in tablets was developed and validated. This method involves separation of the components by TLC on precoated silica gel G60F254 plate with solvent system of n-butanol, toluene and ammonia (8.5:0.5:1 v/v/v) and detection was carried out densitometrically using a UV detector at 288 nm in absorbance mode. This system was found to give compact spots for rabeprazole (Rf value of 0.23 0.02) and for itopride hydrochloride (Rf value of 0.75±0.02). Linearity was found to be in the range of 40-200 ng/spot and 300-1500 ng/spot for rabeprazole and itopride hydrochloride. The limit of detection and limit of quantification for rabeprazole were 10 and 20 ng/spot and for itopride hydrochloride were 50 and 100 ng/spot, respectively. The method was found to be beneficial for the routine analysis of combined dosage form. PMID:20046748

Influence of hypergravity on fish inner ear otoliths: II. Incorporation of calcium and kinetotic behaviour.

PubMed

Beier, M; Anken, R H; Rahmann, H

2002-01-01

Larval siblings of cichlid fish (Oreochromis mossambicus) were subjected to hypergravity (hg; 3 g, 14 days) during development. Following the transfer to 1 g (i.e., stopping the centrifuge) they were separated into normally and kinetotically swimming individuals (the latter performed spinning movements). During hg, the animals were maintained in aquarium water containing alizarin-complexone (AC), a fluorescent calcium tracer. Densitometric measurements of AC uptake into inner ear otoliths (optical density of AC/micrometers2) revealed that the kinetotic individuals had incorporated significantly more AC/calcium than the normally behaving fish. Since the amount of otolithic calcium can be taken as an approximation for otolith weight, the present results indicate that the otoliths of kinetotically swimming samples were heavier than those of the normally behaving larvae, thus exhibiting a higher absolute weight asymmetry of the otoliths between the right vs. the left side of the body. This supports an earlier concept according to which otolith (or statolith) asymmetry is the cause for kinetoses such as human static space sickness. c2002 COSPAR. Published by Elsevier Science Ltd. All rights reserved.

Transient regional osteoporosis.

PubMed

Cano-Marquina, Antonio; Tarín, Juan J; García-Pérez, Miguel-Ángel; Cano, Antonio

2014-04-01

Transient regional osteoporosis (TRO) is a disease that predisposes to fragility fracture in weight bearing joints of mid-life women and men. Pregnant women may also suffer the process, usually at the hip. The prevalence of TRO is lower than the systemic form, associated with postmenopause and advanced age, but may be falsely diminished by under-diagnosis. The disease may be uni- or bilateral, and may migrate to distinct joints. One main feature of TRO is spontaneous recovery. Pain and progressive limitation in the functionality of the affected joint(s) are key symptoms. In the case of the form associated with pregnancy, difficulties in diagnosis derive from the relatively young age at presentation and from the clinical overlapping with the frequent aches during gestation. Densitometric osteoporosis in the affected region is not always present, but bone marrow edema, with or without joint effusion, is detected by magnetic resonance. There are not treatment guidelines, but the association of antiresorptives to symptomatic treatment seems to be beneficial. Surgery or other orthopedic interventions can be required for specific indications, like hip fracture, intra-medullary decompression, or other. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Stability of catechins in green tea nutraceutical products: application of solid phase extraction-thin layer chromatography densitometry.

PubMed

Abd-Elsalam, Heba-Alla H; Al-Ghobashy, Medhat A; Zaazaa, Hala E; Ibrahim, Mohamed A

2014-08-01

Epigallocatechin gallate (EGCG) is a powerful antioxidant and commonly used nutraceutical. Accelerated stability of EGCG in tablet formulations was investigated. LLE and SPE were employed for sample clean-up and enrichment of EGCG over caffeine. Samples were analysed after spiking with fixed concentration of gallic acid (GA), in order to verify reproducibility of analysis. A TLC-densitometric assay was developed and validated for determination of % loss EGCG. EGCG, GA and caffeine were resolved with Rf values 0.54, 0.69 and 0.80, respectively. LC-MS/MS was used to verify identity and purity of the EGCG band. Determination was carried out over a concentration range of 0.50-5.00μg/band and 0.20-2.40μg/band for GA and caffeine, respectively. Results showed significant reduction in EGCG content after one, three and six months: 24.00%, 28.00% and 52.00% respectively. Results continue to demonstrate that stability of nutraceutical products should be investigated in-depth using industry-oriented protocols before granting marketing authorisation. Copyright © 2014 Elsevier Ltd. All rights reserved.

Enhanced healing of full-thickness burn wounds using di-rhamnolipid

PubMed Central

Stipcevic, Tamara; Piljac, Ante; Piljac, Goran

2006-01-01

The aim of this study was to investigate the properties of di-rhamnolipid [α-L-rhamnopyranosyl-(1–2)-α-L-rhamnopyranosyl-3-hydroxydecanoyl-3-hydroxydecanoic acid, also referred to as di-rhamnolipid BAC-3] relating to the process of cutaneous wound healing. Di-rhamnolipid was prepared in a eucerin ointment and applied topically on full-thickness burn wounds in normal Sprague–Dawley rats covering 5% of the total body surface area. The rate of wound closure was measured over the period of 45 days. The collagen content was evaluated microscopically, by performing densitometric analysis on Verhoeff’s stained histopathological slides of wound biopsies taken at the end of 45th day of di-rhamnolipid treatment. Di-rhamnolipid toxicity was assessed with the subcutaneous multi-dose study in Swiss–Webster mice. The treatment of full-thickness-burn wounds with topical 0.1% di-rhamnolipid accelerated the closure of wounds on day 21 of the treatment by 32% compared to the control ( p < 0.05). On day 35, the wounds closed in all animals-treated with 0.1% di-rhamnolipid ointment while some rats in the control group had open wounds on days 35 and even 45. Histologic comparisons have shown that di-rhamnolipid significantly decreased collagen content in burn wounds (47.5%, p < 0.05) as compared to the vehicle-treated (control) wounds. Di-rhamnolipid was well-tolerated. The results of this study raise the possibility of potential efficacy of di-rhamnolipid in accelerating normal wound healing and perhaps in overcoming defects associated with healing failure in chronic wounds. PMID:16380213

CHARACTERISTICS OF RAT LIVER EXPOSED TO NANOPARTICLES OF LEAD COMPOUNDS.

PubMed

Omelchuk, S; Aleksijchuk, V; Sokurenko, L; Blagaia, A; Prudchenko, S

2016-12-01

In recent times, the lead becomes great importance in environmental pollution, including its nanoparticles. In the literature, there is little data on the changes in the liver after the exposure with lead nanoparticles. The purpose of this study was the identification and determination of macroscopic, microscopic, and biochemical changes of the structural elements of the rat's liver exposed to the action of lead compounds. The study was carried out on 60 male Wistar rats. The first and second groups of animals were intraperitoneally injected with colloidal solution of nanoparticles of Lead Sulfide size of 10 nm and 30 nm, and the third group was intraperitoneally injected with a solution of nitrate lead. Macroscopic, histological, histochemical, biochemical methods and gas chromatography were used to identify the changes of fatty acids metabolism. The experiment has found that body weights of animals in all tested groups were decreased after 6 weeks of lead nanoparticles injection, while relative liver weight was increased. Levels of total lipids and cholesterol, total protein and albumin in the blood serum in study groups have decreased, and the level of triglycerides and glucose have increased. Moderate dystrophic changes were observed in the histological examinations of the liver, and this was confirmed by morphometric and densitometric parameters. Changes of fatty acid composition of lipids of the liver exposed to nanoparticles were the result of increasing arachidonic fatty acid content and reduction of the stearic fatty acid content. Thus, it has been proven by the experiment that the effect of lead nanoparticles depends on their size.

A laboratory evaluation of four quality control devices for radiographic processing.

PubMed

Rushton, V E; Horner, K

1994-08-01

Quality assurance programmes for radiographic processing traditionally employ expensive sensitometric and densitometric techniques. However cheap and simple devices for monitoring radiographic processing are available. The aim of this study was to make a comparison of four such devices in terms of their ability to detect variations in radiographic density of clinical significance. Three of the devices are commercially available while the fourth is easily manufactured from waste materials. Ideal bitewing exposure times were selected for four different kilovoltage/film speed combinations. Phantom bitewing radiographs, exposed using these exposure times, were processed using a variety of times and developer temperatures to simulate variations in radiographic quality due to inadequate processing conditions. Test films, produced using the four monitoring devices, were exposed and processed under identical conditions. The phantom bitewings were judged to have 'acceptable' quality when the optical density of that part of the film not showing calcified structures was within +/- 0.5 of that of the film processed under optimal conditions. The efficacy of the monitoring devices in indicating the adequacy of processing was assessed by a comparison of their readings with those made from the phantom bitewings. None of the monitoring devices was ideal for all the kilovoltage/film speed combinations tested, but the homemade device proved to be the most generally effective. We conclude that guidelines to dentists on radiographic quality assurance should include reference to and details of this simple device.

Quantitative Determination of L-DOPA in Seeds of Mucuna Pruriens Germplasm by High Performance Thin Layer Chromatography

PubMed Central

Raina, Archana P.; Khatri, Renu

2011-01-01

Mucuna pruriens Linn. is an important medicinal plant used for treatment of Parkinson's disease and many others in ancient Indian medical system. L-DOPA extracted from seeds of Mucuna is a constituent of more than 200 indigenous drug formulations and is more effective as drug than the synthetic counterpart. A densitometric high performance thin-layer chromatographic (HPTLC) method was developed for quantification of L-DOPA content present in the seeds extract. The method involves separation of L-DOPA on precoated silica gel 60 GF254 HPTLC plates using a solvent system of n-butanol-acetic-acid-water (4:1:1, v/v) as the mobile phase. Quantification was done at 280 nm using absorbance reflectance mode. Linearity was found in the concentration range of 100 to 1000 ng/spot with the correlation coefficient value of 0.9980. The method was validated for accuracy, precision and repeatability. Mean recovery was 100.89%. The LOD and LOQ for L-DOPA determination were found to be 3.41 ng/spot and 10.35 ng/spot respectively. The proposed HPTLC method was found to be precise, specific and accurate for quantitative determination of L-DOPA. It can be used for rapid screening of large germplasm collections of Mucuna pruriens for L-DOPA content. The method was used to study variation in fifteen accessions of Mucuna germplasm collected from different geographical regions. PMID:22707835

Quantitative Determination of L-DOPA in Seeds of Mucuna Pruriens Germplasm by High Performance Thin Layer Chromatography.

PubMed

Raina, Archana P; Khatri, Renu

2011-07-01

Mucuna pruriens Linn. is an important medicinal plant used for treatment of Parkinson's disease and many others in ancient Indian medical system. L-DOPA extracted from seeds of Mucuna is a constituent of more than 200 indigenous drug formulations and is more effective as drug than the synthetic counterpart. A densitometric high performance thin-layer chromatographic (HPTLC) method was developed for quantification of L-DOPA content present in the seeds extract. The method involves separation of L-DOPA on precoated silica gel 60 GF(254) HPTLC plates using a solvent system of n-butanol-acetic-acid-water (4:1:1, v/v) as the mobile phase. Quantification was done at 280 nm using absorbance reflectance mode. Linearity was found in the concentration range of 100 to 1000 ng/spot with the correlation coefficient value of 0.9980. The method was validated for accuracy, precision and repeatability. Mean recovery was 100.89%. The LOD and LOQ for L-DOPA determination were found to be 3.41 ng/spot and 10.35 ng/spot respectively. The proposed HPTLC method was found to be precise, specific and accurate for quantitative determination of L-DOPA. It can be used for rapid screening of large germplasm collections of Mucuna pruriens for L-DOPA content. The method was used to study variation in fifteen accessions of Mucuna germplasm collected from different geographical regions.

Comparative proteomic analysis of the ribosomes in 5-fluorouracil resistance of a human colon cancer cell line using the radical-free and highly reducing method of two-dimensional polyacrylamide gel electrophoresis.

PubMed

Kimura, Kosei; Wada, Akira; Ueta, Masami; Ogata, Akihiko; Tanaka, Satoru; Sakai, Akiko; Yoshida, Hideji; Fushitani, Hideo; Miyamoto, Akiko; Fukushima, Masakazu; Uchiumi, Toshio; Tanigawa, Nobuhiko

2010-11-01

Many auxiliary functions of ribosomal proteins (r-proteins) have received considerable attention in recent years. However, human r-proteins have hardly been examined by proteomic analysis. In this study, we isolated ribosomal particles and subsequently compared the proteome of r-proteins between the DLD-1 human colon cancer cell line and its 5-fluorouracil (5-FU)-resistant sub-line, DLD-1/5-FU, using the radical-free and highly reducing method of two-dimensional polyacrylamide gel electrophoresis, which has a superior ability to separate basic proteins, and we discuss the role of r-proteins in 5-FU resistance. Densitometric analysis was performed to quantify modulated proteins, and protein spots showing significant changes were identified by employing matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. Three basic proteins (L15, L37 and prohibitin) which were significantly modulated between DLD-1 and DLD-1/5-FU were identified. Two proteins, L15 and L37, showed down-regulated expression in DLD-1/5-FU in comparison to DLD-1. Prohibitin, which is not an r-protein and is known to be localized in the mitochondria, showed up-regulated expression in DLD-1/5-FU. These 3 proteins may be related to 5-FU resistance.

Genetic linkage analysis using pooled DNA and infrared detection of tailed STRP primer patterns

NASA Astrophysics Data System (ADS)

Oetting, William S.; Wildenberg, Scott C.; King, Richard A.

1996-04-01

The mapping of a disease locus to a specific chromosomal region is an important step in the eventual isolation and analysis of a disease causing gene. Conventional mapping methods analyze large multiplex families and/or smaller nuclear families to find linkage between the disease and a chromosome marker that maps to a known chromosomal region. This analysis is time consuming and tedious, typically requiring the determination of 30,000 genotypes or more. For appropriate populations, we have instead utilized pooled DNA samples for gene mapping which greatly reduces the amount of time necessary for an initial chromosomal screen. This technique assumes a common founder for the disease locus of interest and searches for a region of a chromosome shared between affected individuals. Our analysis involves the PCR amplification of short tandem repeat polymorphisms (STRP) to detect these shared regions. In order to reduce the cost of genotyping, we have designed unlabeled tailed PCR primers which, when combined with a labeled universal primer, provides for an alternative to synthesizing custom labeled primers. The STRP pattern is visualized with an infrared fluorescence based automated DNA sequencer and the patterns quantitated by densitometric analysis of the allele pattern. Differences in the distribution of alleles between pools of affected and unaffected individuals, including a reduction in the number of alleles in the affected pool, indicate the sharing of a region of a chromosome. We have found this method effective for markers 10 - 15 cM away from the disease locus for a recessive genetic disease.

Stability-indicating methods for the determination of racecadotril in the presence of its degradation products.

PubMed

Mohamed, Afaf O; Fouad, Manal M; Hasan, Mona M; Abdel Razeq, Sawsan A; Elsherif, Zeinab A

2009-12-01

Three stability-indicating methods were developed for the determination of racecadotril (RCT) in the presence of its alkaline degradation products. The first was an HPLC method in which efficient chromatographic separation was achieved on a C18 analytical column and a mobile phase of acetonitrile-methanol-water-acetic acid (52:28:20:0.1, v/v/v/v). Linearity was obtained in the range of 4-40 microg/mL with mean accuracy of 99.5 +/- 0.88%. The second method was a densitometric evaluation of thin-layer chromatograms of the drug using a mobile phase of isopropanol-ammonia (33%)-n-hexane (9:0.5:20, v/v/v). The chromatograms were scanned at 232 nm, a wavelength at which RCT can be readily separated from its degradation products and determined in the range of 2-20 microg per spot with mean accuracy of 99.5 +/- 0.56%. The third method is based on the use of first-derivative spectrophotometry (D1) at 240 nm, and the drug was determined in the range of 5-40 microg/mL with mean accuracy of 99.2 +/- 1.02%. The three methods provided satisfactory recovery of the intact drug (100.8 +/- 0.82, 100.4 +/- 0.55, and 99.9 +/- 0.72%, respectively) in the presence of up to 90% of its degradation products. Determination was also successful when analyzing RCT in a formulation in the form of acetorphan packets. Results were statistically analyzed and found to be in accordance with those given by a reported method.

Development and Validation of a High-Performance Thin-Layer Chromatographic Method for the Simultaneous Determination of Two Binary Mixtures Containing Ketorolac Tromethamine with Phenylephrine Hydrochloride and with Febuxostat

PubMed Central

El Yazbi, Fawzy A.; Hassan, Ekram M.; Khamis, Essam F.; Ragab, Marwa A.A.; Hamdy, Mohamed M.A.

2016-01-01

A validated and highly selective high-performance thin-layer chromatography (HPTLC) method was developed for the determination of ketorolac tromethamine (KTC) with phenylephrine hydrochloride (PHE) (Mixture 1) and with febuxostat (FBX) (Mixture 2) in bulk drug and in combined dosage forms. The proposed method was based on HPTLC separation of the drugs followed by densitometric measurements of their spots at 273 and 320 nm for Mixtures 1 and 2, respectively. The separation was carried out on Merck HPTLC aluminum sheets of silica gel 60 F254 using chloroform–methanol–ammonia (7:3:0.1, v/v) and (7.5:2.5:0.1, v/v) as mobile phase for KTC/PHE and KTC/FBX mixtures, respectively. Linear regression lines were obtained over the concentration ranges 0.20–0.60 and 0.60–1.95 µg band−1 for KTC and PHE (Mixture 1), respectively, and 0.10–1.00 and 0.25–2.50 µg band−1 for KTC and FBX (Mixture 2), respectively, with correlation coefficients higher than 0.999. The method was successfully applied to the analysis of the two drugs in their synthetic mixtures and in their dosage forms. The mean percentage recoveries were in the range of 98–102%, and the RSD did not exceed 2%. The method was validated according to ICH guidelines and showed good performances in terms of linearity, sensitivity, precision, accuracy and stability. PMID:26847918

Comparative evaluation of bovine derived hydroxyapatite and synthetic hydroxyapatite graft in bone regeneration of human maxillary cystic defects: a clinico-radiological study.

PubMed

Kattimani, Vivekanand S; Chakravarthi, Srinivas P; Neelima Devi, K Naga; Sridhar, Meka S; Prasad, L Krishna

2014-01-01

Bone grafts are frequently used in the treatment of bone defects. Bone harvesting can cause postoperative complications and sometimes does not provide a sufficient quantity of bone. Therefore, synthetic biomaterials have been investigated as an alternative to autogenous bone grafts. The aim of this study was to evaluate and compare bovine derived hydroxyapatite (BHA) and synthetic hydroxyapatite (SHA) graft material as bone graft substitute in maxillary cystic bony defects. Patients were analyzed by computerized densitometric study and digital radiography. In this study, 12 patients in each group were included randomly after clinical and radiological evaluation. The integration of hydroxyapatite was assessed with mean bone density, surgical site margin, and radiological bone formation characteristics, of the successful graft cases using computer densitometry and radio-visiograph. Statistical analysis was carried out using Mann-Whitney U-test, Wilcoxon matched pairs test and paired t-test. By the end of 24 th week, the grafted defects radiologically and statistically showed similar volumes of bone formation. However, the significant changes observed in the formation of bone and merging of material and surgical site margin at 1 st week to 1 st month. The results were significant and correlating with all the parameters showing the necessity of the grafting for early bone formation. However, the bone formation pattern is different in both BHA and SHA group at 3 rd month interval with significant P value. Both BHA and SHA graft materials are biocompatible for filling bone defects, showing less resorption and enhanced bone formation with similar efficacy. Our study showed maximum bone healing within 12 weeks of grafting of defects. The BHA is economical; however, price difference between the two is very nominal.

Morphological and histomorphometric evaluation of the ventral rectus sheath of the rectus abdominis muscle, fascia lata and pectoral fascia. The beginning of a morphological information bank of human fascias.

PubMed

Morales-Avalos, Rodolfo; Soto-Domínguez, Adolfo; García-Juárez, Jaime; Cardenas-Serna, Marcela; Esparza-Hernández, Claudia N; Carreño-Salcedo, Sofía Alejandra; Montes-de-Oca-Luna, Roberto; Loera-Arias, María de Jesús; Saucedo-Cárdenas, Odila; Elizondo-Omaña, Rodrigo E; Guzmán-López, Santos

2017-03-01

The aim of this study was to characterize and compare the morphological and histomorphometric characteristics of the pectoral fascia, fascia lata and ventral rectus sheath. Twenty cadaveric samples of these fascias were analyzed and stained with hematoxylin and eosin, orcein, Van Gieson, Masson's trichrome and Verhoeff¨s stain (1200 slides in total). Morphological evaluation, semiquantitative, morphometric and microdensitometric analysis of elastic fibers present in each of the tissues and a morphometrical analysis of tissue thickness were performed. The mean value of the pectoral fascia thickness was 612±68.13 μm; 84±246 μm for the fascia lata and 584±92 μm for the ventral rectus sheath. The area occupied by the elastic fibers in the pectoral fascia was 12.24±5.84%; 6,54±3.85% for the fascia lata and 11.11±5.26% for the ventral rectus sheath. There were no statistically significant differences when comparing the mean values between the pectoral fascia and the ventral rectus sheath (p=0.07). There were statistically significant differences when comparing the fascia lata to the pectoral fascia and the ventral rectus sheath (p≤0.001). This study reports other morphological characteristics not described in previous histological studies of the analyzed tissues. The results of the morphometric and densitometric analysis in this study reveal that the fascia lata has the fewest elastic fibers of all the tissues analyzed, and the pectoral fascia has the most. These results will be useful for the beginning of a morphological information bank of human fascias.

[Prevalence of osteoporosis, estimation of probability of fracture and bone metabolism study in patients with newly diagnosed prostate cancer in the health area of Lugo].

PubMed

Miguel-Carrera, Jonatan; García-Porrua, Carlos; de Toro Santos, Francisco Javier; Picallo-Sánchez, Jose Antonio

2018-03-01

To study the prevalence of osteoporosis and fracture probability in patients diagnosed with prostate cancer. Observational descriptive transversal study. SITE: Study performed from Primary Care of Lugo in collaboration with Rheumatology and Urology Services of our referral hospital. Patients diagnosed with prostate cancer without bone metastatic disease from January to December 2012. Epidemiologic, clinical, laboratory and densitometric variables involved in osteoporosis were collected. The likelihood of fracture was estimated by FRAX ® Tool. Eighty-three patients met the inclusion criteria. None was excluded. The average age was 67 years. The Body Mass Index was 28.28. Twenty-five patients (30.1%) had previous osteoporotic fractures. Other prevalent risk factors were alcohol (26.5%) and smoking (22.9%). Eighty-two subjects had vitamin D below normal level (98.80%). Femoral Neck densitometry showed that 8.9% had osteoporosis and 54% osteopenia. The average fracture risk in this population, estimated by FRAX ® , was 2.63% for hip fracture and 5.28% for major fracture. Cut level for FRAX ® major fracture value without DXA >5% and ≥7.5% proposed by Azagra et al. showed 24 patients (28.92%) and 8 patients (9.64%) respectively. The prevalence of osteoporosis in this population was very high. The more frequent risk factors associated with osteoporosis were: previous osteoporotic fracture, alcohol consumption, smoking and family history of previous fracture. The probability of fracture using femoral neck FRAX ® tool was low. Vitamin D deficiency was very common (98.8%). Copyright © 2017 Elsevier España, S.L.U. All rights reserved.

Imaging Phenotype of Occupational Endotoxin-Related Lung Function Decline.

PubMed

Lai, Peggy S; Hang, Jing-Qing; Zhang, Feng-Ying; Sun, J; Zheng, Bu-Yong; Su, Li; Washko, George R; Christiani, David C

2016-09-01

Although occupational exposures contribute to a significant proportion of obstructive lung disease, the phenotype of obstructive lung disease associated with work-related organic dust exposure independent of smoking remains poorly defined. We identified the relative contributions of smoking and occupational endotoxin exposure to parenchymal and airway remodeling as defined by quantitative computed tomography (CT). The Shanghai Textile Worker Study is a longitudinal study of endotoxin-exposed cotton workers and endotoxin-unexposed silk workers that was initiated in 1981. Spirometry, occupational endotoxin exposure, and smoking habits were assessed at 5-year intervals. High-resolution computed tomography (CT) was performed in 464 retired workers in 2011, along with quantitative lung densitometric and airway analysis. Significant differences in all CT measures were noted across exposure groups. Occupational endotoxin exposure was associated with a decrease (-1.3%) in percent emphysema (LAAI-950), a 3.3-Hounsfield unit increase in 15th percentile density, an 18.1-g increase in lung mass, and a 2.3% increase in wall area percent. Current but not former smoking was associated with a similar CT phenotype. Changes in LAAI-950 were highly correlated with 15th percentile density (correlation -1.0). Lung mass was the only measure associated with forced expiratory volume in 1 sec (FEV1) decline, with each 10-g increase in lung mass associated with an additional loss (-6.1 mL) of FEV1 (p = 0.001) between 1981 and 2011. There are many similarities between the effects of occupational endotoxin exposure and those of tobacco smoke exposure on lung parenchyma and airway remodeling. The effects of occupational endotoxin exposure appear to persist even after the cessation of exposure. LAAI-950 may not be a reliable indicator of emphysema in subjects without spirometric impairment. Lung mass is a CT-based biomarker of accelerated lung function decline. Lai PS, Hang J, Zhang F, Sun J, Zheng BY, Su L, Washko GR, Christiani DC. 2016. Imaging phenotype of occupational endotoxin-related lung function decline. Environ Health Perspect 124:1436-1442; http://dx.doi.org/10.1289/EHP195.

Development and validation of a normal-phase high-performance thin layer chromatographic method for the analysis of sulfamethoxazole and trimethoprim in co-trimoxazole tablets.

PubMed

Shewiyo, D H; Kaale, E; Risha, P G; Dejaegher, B; Smeyers-Verbeke, J; Vander Heyden, Y

2009-10-16

Pneumocystis carinii pneumonia (PCP) is often the ultimate mortal cause for immunocompromised individuals, such as HIV/AIDS patients. Currently, the most effective medicine for treatment and prophylaxis is co-trimoxazole, a synergistic combination of sulfamethoxazole (SMX) and trimethoprim (TMP). In order to ensure a continued availability of high quality co-trimoxazole tablets within resource-limited countries, Medicines Regulatory Authorities must perform quality control of these products. However, most pharmacopoeial methods are based on high-performance liquid chromatographic (HPLC) methods. Because of the lack of equipment, the Tanzania Food and Drugs Authority (TFDA) laboratory decided to develop and validate an alternative method of analysis based on the TLC technique with densitometric detection, for the routine quality control of co-trimoxazole tablets. SMX and TMP were separated on glass-backed silica gel 60 F(254) plates in a high-performance thin layer chromatograph (HPTLC). The mobile phase was comprised of toluene, ethylacetate and methanol (50:28.5:21.5, v:v:v). Detection wavelength was 254 nm. The R(f) values were 0.30 and 0.61 for TMP and SMX, respectively. This method was validated for linearity, precision, trueness, specificity and robustness. Cochran's criterion test indicated homoscedasticity of variances for the calibration data. The F-tests for lack-of-fit indicated that straight lines were adequate to describe the relationship between spot areas and concentrations for each compound. The percentage relative standard deviations for repeatability and time-different precisions were 0.98 and 1.32, and 0.83 and 1.64 for SMX and TMP, respectively. Percentage recovery values were 99.00%+/-1.83 and 99.66%+/-1.21 for SMX and TMP, respectively. The method was found to be robust and was then successfully applied to analyze co-trimoxazole tablet samples.

Phototoxic Effect of Topical Fluoroquinolones Administered Before Corneal Crosslinking in a Murine Model.

PubMed

Reviglio, Victor E; Osaba, Matias; Sambuelli, Gabriela; Kuo, Irene C

2017-03-01

Corneal crosslinking by UV light (UV-CXL) has become a popular treatment for keratoconus and corneal ectasia. Fluoroquinolones (FQs), commonly administered topically before UV-CXL, are known to be phototoxic to the skin and lens. The purpose of this study was to investigate phototoxic effects of topical FQ treatment on murine corneas before UV-CXL, in which the corneal epithelium was kept intact. Murine corneas were treated with various antibiotics with or without riboflavin before UV-CXL. At 24 h, the animals were sacrificed, and the corneas were analyzed for histologic evidence of inflammation and apoptosis and for expression of apoptosis markers BAX and caspases 3 and 9 and for expression of matrix metalloproteinase 9 (MMP-9). Spectrofluorometric analysis was performed. Corneas treated with topical FQ with or without riboflavin before UV-CXL showed mild corneal stromal inflammation, apoptosis by both terminal deoxynucleotidyl transferase dUTP nick end labeling staining and increased expression of BAX gene and caspases 3 and 9 by densitometric analysis. Untreated corneas, corneas treated with azithromycin before UV-CXL, and corneas undergoing UV-CXL without any antibiotic or riboflavin pretreatment showed normal histology, no staining for apoptosis, and no increased production of apoptosis markers by polymerase chain reaction. The phototoxic effects of FQs on the cornea may lead surgeons to consider another antibiotic class for prophylaxis against infectious keratitis in UV-CXL. These effects, along with the known cytotoxic effects of FQs independent of UV radiation, may contribute to some of the complications of corneal UV-CXL. Dosage studies may be warranted.

Comparative efficacy of photo-activated disinfection and calcium hydroxide for disinfection of remaining carious dentin in deep cavities: a clinical study

PubMed Central

Sharma, Sidhartha; Shah, Naseem

2014-01-01

Objectives To comparatively evaluate the efficacy of photo-activated disinfection (PAD), calcium hydroxide (CH) and their combination on the treatment outcome of indirect pulp treatment (IPT). Materials and Methods Institutional ethical clearance and informed consent of the patients were taken. The study was also registered with clinical registry of India. Sixty permanent molars exhibiting deep occlusal carious lesion in patients with the age range of 18 - 22 yr were included. Clinical and radiographic evaluation and set inclusion and exclusion criteria's were followed. Gross caries excavation was accomplished. In group I (n = 20) PAD was applied for sixty seconds. In group II (n = 20), CH was applied to the remaining carious dentin, while in group III (n = 20), PAD application was followed by CH placement. The teeth were permanently restored. They were clinically and radiographically followed-up at 45 day, 6 mon and 12 mon. Relative density of the remaining affected dentin was measured by 'Radiovisiography (RVG) densitometric' analysis. Results Successful outcome with an increase in radiographic grey values were observed in all three groups. However, on inter-group comparison, this change was not significant (p > 0.05). Conclusions PAD and CH both have equal disinfection efficacy in the treatment of deep carious dentin. PAD alone is as effective for treatment of deep carious lesion as calcium hydroxide and hence can be used as an alternative to CH. They can be used independently in IPT, since combining both does not offer any additional therapeutic benefits. PMID:25110643

Effect of Hypergravity on Carbonanhydrase Reactivity in inner Ear Ioncytes of developing Cichlid Fish

NASA Astrophysics Data System (ADS)

Beier, M.; Anken, R.; Rahmann, H.

It has been shown earlier that hypergravity slows down inner ear otolith growth in developing fish. Otolith growth in terms of mineralisation mainly depends on the enzyme carboanhydrase (CAH), which is responsible for the provision of the pH- value necessary for calcium carbonate deposition and thus also is presumed to play a prominent role in Ménière's disease (a sensory - motor disorder inducing vertigo and kinetosis). Larval siblings of cichlid fish (Oreochromis mossambicus) were subjected to hypergravity (3g; 6 hours) during development and separated into normally and kinetotically swimming individuals following the transfer to 1g (i.e., stopping the centrifuge; kinetotically behaving fish performed spinning movements). Subsequently, CAH was histochemically demonstrated in inner ear ionocytes (cells involved in the endolymphatic ion exchange) and enzyme reactivity was determined densitometrically. The results showed that CAH-reactivity was significantly increased in normally behaving hyper-g specimens as compared to controls kept at 1g, whereas no difference in enzyme reactivity was evident between the controls and kinetotically behaving fish. On the background of earlier studies, according to which (1) hypergravity induces a decrease of otolith growth and (2) the otolithic calcium incorporation (visualized using the calcium -tracer alizarin complexone) of kinetotically swimming hyper - g fish was lower as compared to normally behaving hyper - g animals, the present study strongly supports the concept that an increase in CAH-reactivity may result in a decrease of otolithic calcium deposition. The mechanism regulating CAH-activity hitherto remains to be determined. Acknowledgement: This work was financially supported by the German Aerospace Center (DLR) (FKZ: 50 WB 9997).

The effects of once-weekly teriparatide on hip structure and biomechanical properties assessed by CT.

PubMed

Ito, M; Oishi, R; Fukunaga, M; Sone, T; Sugimoto, T; Shiraki, M; Nishizawa, Y; Nakamura, T

2014-03-01

Once-weekly administration of 56.5 μg teriparatide improved cortical bone parameters and biomechanical parameters at the proximal femur by CT geometry analysis. The aim of this study was to evaluate the effects of weekly administration of teriparatide [human PTH (1-34)] on bone geometry, volumetric bone mineral density (vBMD), and parameters of bone strength at the proximal femur which were longitudinally investigated using computed tomography (CT). The subjects were a subgroup of a recent, randomly assigned, double-blind study (578 subjects) comparing the anti-fracture efficacy of a once-weekly subcutaneous injection of 56.5 μg teriparatide with placebo (TOWER trial). Sixty-six ambulatory postmenopausal women with osteoporosis were enrolled at 15 study sites having multi-detector row CT, and included women injected with teriparatide (n = 29, 74.2 ± 5.1 years) or with placebo (n = 37, 74.8 ± 5.3 years). CT data were obtained at baseline and follow-up scans were performed at 48 and 72 weeks. The data were analyzed to obtain cross-sectional densitometric, geometric, and biomechanical parameters including the section modulus (SM) and buckling ratio (BR) of the femoral neck, inter-trochanter, and femoral shaft. We found that once-weekly teriparatide increased cortical thickness/cross-sectional area (CSA) and total area, and improved biomechanical properties (i.e., decreasing BR) at the femoral neck and shaft. Teriparatide did not change the cortical perimeter. Our longitudinal analysis of proximal femur geometry by CT revealed that once-weekly administration of 56.5 μg teriparatide improved cortical bone parameters at the femoral neck and shaft and also improved biomechanical parameters.

Reduction of lymphocyte G protein-coupled receptor kinase-2 (GRK2) after exercise training predicts survival in patients with heart failure.

PubMed

Rengo, Giuseppe; Galasso, Gennaro; Femminella, Grazia D; Parisi, Valentina; Zincarelli, Carmela; Pagano, Gennaro; De Lucia, Claudio; Cannavo, Alessandro; Liccardo, Daniela; Marciano, Caterina; Vigorito, Carlo; Giallauria, Francesco; Ferrara, Nicola; Furgi, Giuseppe; Filardi, Pasquale Perrone; Koch, Walter J; Leosco, Dario

2014-01-01

Increased cardiac G protein-coupled receptor kinase-2 (GRK2) expression has a pivotal role at inducing heart failure (HF)-related β-adrenergic receptor (βAR) dysfunction. Importantly, abnormalities of βAR signalling in the failing heart, including GRK2 overexpression, are mirrored in circulating lymphocytes and correlate with HF severity. Exercise training has been shown to exert several beneficial effects on the failing heart, including normalization of cardiac βAR function and GRK2 protein levels. In the present study, we evaluated whether lymphocyte GRK2 levels and short-term changes of this kinase after an exercise training programme can predict long-term survival in HF patients. For this purpose, we prospectively studied 193 HF patients who underwent a 3-month exercise training programme. Lymphocyte GRK2 protein levels, plasma N-terminal pro-brain natriuretic peptide, and norepinephrine were measured at baseline and after training along with clinical and functional parameters (left ventricular ejection fraction, NYHA class, and peak-VO2). Cardiac-related mortality was evaluated during a mean follow-up period of 37 ± 20 months. Exercise was associated with a significant reduction of lymphocyte GRK2 protein levels (from 1.29 ± 0.52 to 1.16 ± 0.65 densitometric units, p < 0.0001). Importantly, exercise related changes of GRK2 (delta values) robustly predicted survival in our study population. Interestingly, HF patients who did not show reduced lymphocyte GRK2 protein levels after training presented the poorest outcome. Our data offer the first demonstration that changes of lymphocyte GRK2 after exercise training can strongly predict outcome in advanced HF.

Extracorporeal shock wave therapy in treatment of delayed bone-tendon healing.

PubMed

Wang, Lin; Qin, Ling; Lu, Hong-bin; Cheung, Wing-hoi; Yang, Hu; Wong, Wan-nar; Chan, Kai-ming; Leung, Kwok-sui

2008-02-01

Extracorporeal shock wave therapy is indicated for treatment of chronic injuries of soft tissues and delayed fracture healing and nonunion. No investigation has been conducted to study the effect of shock wave on delayed healing at the bone-tendon junction. Shock wave promotes osteogenesis, regeneration of fibrocartilage zone, and remodeling of healing tissue in delayed healing of bone-tendon junction surgical repair. Controlled laboratory study. Twenty-eight mature rabbits were used for establishing a delayed healing model at the patella-patellar tendon complex after partial patellectomy and then divided into control and shock wave groups. In the shock wave group, a single shock wave treatment was given at week 6 postoperatively to the patella-patellar tendon healing complex. Seven samples were harvested at week 8 and 7 samples at week 12 for radiologic, densitometric, histologic, and mechanical evaluations. Radiographic measurements showed 293.4% and 185.8% more new bone formation at the patella-patellar tendon healing junction in the shock wave group at weeks 8 and 12, respectively. Significantly better bone mineral status was found in the week 12 shock wave group. Histologically, the shock wave group showed more advanced remodeling in terms of better alignment of collagen fibers and thicker and more mature regenerated fibrocartilage zone at both weeks 8 and 12. Mechanical testing showed 167.7% and 145.1% higher tensile load and strength in the shock wave group at week 8 and week 12, respectively, compared with controls. Extracorporeal shock wave promotes osteogenesis, regeneration of fibrocartilage zone, and remodeling in the delayed bone-to-tendon healing junction in rabbits. These results provide a foundation for future clinical studies toward establishment of clinical indication for treatment of delayed bone-to-tendon junction healing.

Beta-galactosidase deficiency in a Korat cat: a new form of feline GM1-gangliosidosis.

PubMed

De Maria, R; Divari, S; Bo, S; Sonnio, S; Lotti, D; Capucchio, M T; Castagnaro, M

1998-09-01

A 7-month-old Korat cat was referred for a slowly progressive neurological disease. Circulating monocytes and lymphocytes showed the presence of single or multiple empty vacuoles and blood leukocytes enzyme assay revealed a very low beta-galactosidase activity level (4.7 nmol/mg per h) as compared to unaffected parents and relatives. Histologically, the cat, euthanized at the owner request at 21 months of age, presented diffuse vacuolization and enlargement of neurons throughout the brain, spinal cord and peripheral ganglia, severe cerebellar neuronal cell loss, and moderate astrocytosis. Stored material was stained with periodic acid-Schiff on frozen sections and with the lectins Ricinus conmmunis agglutinin-I, concanavalin A and wheat germ agglutinin on paraffin-embedded sections. Ultrastructurally, neuronal vacuoles were filled with concentrically whorled lamellae and small membrane-bound vesicles. In the affected cat, beta-galactosidase activity was markedly reduced in brain (18.9%) and liver (33.25%), while total beta-hexosaminidase activity showed a remarkable increase. Quantitation of total gangliosides revealed a 3-fold increase in brain and 1.7-fold in liver of affected cat. High-performance thin layer chromatography (HPTLC) detected a striking increase of GM1-ganglioside. On densitometric analysis of HPTLC bands, the absorption of GM1-ganglioside band was 98.52% of all stained bands (GD1a, GD1b, GT1b). Based on clinical onset, morphological and histochemical features, and biochemical findings, the Korat cat GM1-gangliosidosis is comparable with the human type II (juvenile) form. However, clinical progression, survival time and level of beta-galactosidase deficiency do not completely fit with those of human type II GM1-gangliosidosis. The disease in the Korat cat is also different from other reported forms of feline GM1-gangliosidosis.

Ultrasound Tissue Characterization of Vulnerable Atherosclerotic Plaque

PubMed Central

Picano, Eugenio; Paterni, Marco

2015-01-01

A thrombotic occlusion of the vessel fed by ruptured coronary atherosclerotic plaque may result in unstable angina, myocardial infarction or death, whereas embolization from a plaque in carotid arteries may result in transient ischemic attack or stroke. The atherosclerotic plaque prone to such clinical events is termed high-risk or vulnerable plaque, and its identification in humans before it becomes symptomatic has been elusive to date. Ultrasonic tissue characterization of the atherosclerotic plaque is possible with different techniques—such as vascular, transesophageal, and intravascular ultrasound—on a variety of arterial segments, including carotid, aorta, and coronary districts. The image analysis can be based on visual, video-densitometric or radiofrequency methods and identifies three distinct textural patterns: hypo-echoic (corresponding to lipid- and hemorrhage-rich plaque), iso- or moderately hyper-echoic (fibrotic or fibro-fatty plaque), and markedly hyperechoic with shadowing (calcific plaque). Hypoechoic or dishomogeneous plaques, with spotty microcalcification and large plaque burden, with plaque neovascularization and surface irregularities by contrast-enhanced ultrasound, are more prone to clinical complications than hyperechoic, extensively calcified, homogeneous plaques with limited plaque burden, smooth luminal plaque surface and absence of neovascularization. Plaque ultrasound morphology is important, along with plaque geometry, in determining the atherosclerotic prognostic burden in the individual patient. New quantitative methods beyond backscatter (to include speed of sound, attenuation, strain, temperature, and high order statistics) are under development to evaluate vascular tissues. Although not yet ready for widespread clinical use, tissue characterization is listed by the American Society of Echocardiography roadmap to 2020 as one of the most promising fields of application in cardiovascular ultrasound imaging, offering unique opportunities for the early detection and treatment of atherosclerotic disease. PMID:25950760

Impact of an osteoporosis specialized unit on bone health in breast cancer survivals treated with aromatase inhibitors.

PubMed

Martínez, Purificación; Galve, Elena; Arrazubi, Virginia; Sala, M Ángeles; Fernández, Seila; Pérez, Clara E; Arango, Juan F; Torre, Iñaki

2017-10-11

Considering the increased fracture risk in early breast cancer patients treated with aromatase inhibitors (AI), we assessed the impact of a preventive intervention conducted by a specialized osteoporosis unit on bone health at AI treatment start. Retrospective cohort of postmenopausal women who started treatment with AI after breast cancer surgical/chemotherapy treatment and were referred to the osteoporosis unit for a comprehensive assessment of bone health. Bone densitometry and fracture screening by plain X-ray were performed at the baseline visit and once a year for 5 years. The final record included 130 patients. At AI treatment start, 49% had at least one high-risk factor for fractures, 55% had osteopenia, and 39% osteoporosis. Based on the baseline assessment, 79% of patients initiated treatment with bisphosphonates, 88% with calcium, and 79% with vitamin D. After a median of 65 (50-77) months, 4% developed osteopenia or osteoporosis, and 14% improved their densitometric diagnosis. Fifteen fractures were recorded in 11 (8.5%) patients, all of them receiving preventive treatment (10 with bisphosphonates). During the follow-up period, patients with one or more high-risk factors for fracture showed a greater frequency of fractures (15% vs. 3%) and experienced the first fracture earlier than those without high-risk factors (mean of 99 and 102 months, respectively; P=0.023). The preventive intervention of a specialized unit at the start of AI treatment in breast cancer survivors allows the identification of patients with high fracture risk and may contribute to preventing bone events in these patients. Copyright © 2017 Elsevier España, S.L.U. and Sociedad Española de Reumatología y Colegio Mexicano de Reumatología. All rights reserved.

Influence of long-term hyper-gravity on the reactivity of succinic acid dehydrogenase and NADPH-diaphorase in the central nervous system of fish: a histochemical study

NASA Astrophysics Data System (ADS)

Anken, R. H.; Rahmann, H.

In the course of a densitometric evaluation, the histochemically demonstrated reactivity of succinic acid dehydrogenase (SDH) and of NADPH-diaphorase (NADPHD) was determined in different brain nuclei of two teleost fish (cichlid fish Oreochromis mossambicus, swordtail fish Xiphophorus helleri), which had been kept under 3g hyper-gravity for 8 days. SDH was chosen since it is a rate limiting enzyme of the Krebs cycle and therefore it is regarded as a marker for metabolic and neuronal activity. NADPHD reactivity reflects the activity of nitric oxide synthase. Nitric oxide (NO) is a gaseous intercellular messenger that has been suggested to play a major role in several different in vivo models of neuronal plasticity including learning. Within particular vestibulum-connected brain centers, significant effects of hyper-gravity were obtained, e.g., in the magnocellular nucleus, a primary vestibular relay ganglion of the brain stem octavolateralis area, in the superior rectus subdivision of the oculomotoric nucleus and within cerebellar eurydendroid cells, which in teleosts possibly resemble the deep cerebellar nucleus of higher vertebrates. Non-vestibulum related nuclei did not respond to hypergravity in a significant way. The effect of hyper-gravity found was much less distinct in adult animals as compared to the circumstances seen in larval fish (Anken et al., Adv. Space Res. 17, 1996), possibly due to a development correlated loss of neuronal plasticity.

[Clinical and physiological evaluation of bone changes among astronauts after long-term space flights

NASA Technical Reports Server (NTRS)

Grigoriev, A. I.; Oganov, V. S.; Bakulin, A. V.; Poliakov, V. V.; Voronin, L. I.; Morgun, V. V.; Shnaider, V. S.; Murashko, L. V.; Novikov, V. E.; LeBlank, A.;

Analysis of Bufo arenarum oviductal secretion during the sexual cycle.

PubMed

Crespo, Claudia A; Ramos, Inés; Medina, Marcela F; Fernández, Silvia N

2009-11-01

SummaryBufo arenarum oocytes are oviposited surrounded by jelly coats, one component of the extracellular matrix required for fertilization. The secretion, released to the oviductal lumen, was analysed by SDS-PAGE. The coomassie blue staining evidenced an electrophoretic pattern with molecules ranging between 300 and 19 kDa that showed variations in their secretion profiles during the sexual cycle. In the preovulatory period the densitometric analysis showed the presence of nine peaks with marked predominance of the 74 kDa molecule. Once ovulation has occurred, the jelly coats become arranged around the oocytes during their transit throughout the oviductal pars convoluta (PC), revealing the addition of three proteins only observed during this period, which suggests a differential secretion. Some of these proteins could not diffuse under any extraction treatment, indicating for them a structural or in situ function. Proteins of low molecular mass diffused totally while others showed a partial diffusing capacity. After ovulation a marked decrease in the relative amount of all the proteins released to the lumen, especially the 74 kDa protein, could be detected. During this period, unlike the other stages of the sexual cycle, a differential secretion pattern was observed along the PC. The histochemical analysis performed during the ovulatory period showed the presence of glycoconjugates including both acidic and neutral groups. The present results are in agreement with previous ultrastructural and histochemical studies that describe the role of Bufo arenarum jelly coats in fertilization.

Quantitative computed tomography for the prediction of pulmonary function after lung cancer surgery: a simple method using simulation software.

PubMed

Ueda, Kazuhiro; Tanaka, Toshiki; Li, Tao-Sheng; Tanaka, Nobuyuki; Hamano, Kimikazu

2009-03-01

The prediction of pulmonary functional reserve is mandatory in therapeutic decision-making for patients with resectable lung cancer, especially those with underlying lung disease. Volumetric analysis in combination with densitometric analysis of the affected lung lobe or segment with quantitative computed tomography (CT) helps to identify residual pulmonary function, although the utility of this modality needs investigation. The subjects of this prospective study were 30 patients with resectable lung cancer. A three-dimensional CT lung model was created with voxels representing normal lung attenuation (-600 to -910 Hounsfield units). Residual pulmonary function was predicted by drawing a boundary line between the lung to be preserved and that to be resected, directly on the lung model. The predicted values were correlated with the postoperative measured values. The predicted and measured values corresponded well (r=0.89, p<0.001). Although the predicted values corresponded with values predicted by simple calculation using a segment-counting method (r=0.98), there were two outliers whose pulmonary functional reserves were predicted more accurately by CT than by segment counting. The measured pulmonary functional reserves were significantly higher than the predicted values in patients with extensive emphysematous areas (<-910 Hounsfield units), but not in patients with chronic obstructive pulmonary disease. Quantitative CT yielded accurate prediction of functional reserve after lung cancer surgery and helped to identify patients whose functional reserves are likely to be underestimated. Hence, this modality should be utilized for patients with marginal pulmonary function.

Experiment M-6: Bone Demineralization

NASA Technical Reports Server (NTRS)

Mack, Pauline B.; Vose, George; Vogt, Fred B.; LaChance, Paul A.

1966-01-01

Densitometric evaluations of serial radiographs of "normal" subjects have often shown rather frequent changes in bone mass within relatively short periods of time. For this reason it was decided to make two pre-flight and two post flight radiographs of the Gemini V backup crew. In comparing the changes observed preflight and post flight as the conventional os calcis scanning site between the two crews, it was found that no changes greater than 4 percent were evident in either member of the backup crew. In comparing the changes observed preflight and postflight as the conventional o calcis scanning site between the two crews, it was found that no changes greater than 4 percent were evident in either member of the backup crew. This is in contract to the 15.1 and 8.9 percent losses observed in the prime crew. It has long been known that the skeletal system experiences a general loss of mineral under immobilization or extended bed rest. However, in both Gemini IV and Gemini V studies, bone mass losses were greater in both the os calcis and phalanx than were shown by the TWU bed-rest subjects during the same period of time. Although the bone mass losses in the 8-day Gemini V flight were generally greater than in the 4-day Gemini IV flight, the information to date is still insufficient to conclude that the losses tend to progress linearly with time, or whether a form of physiological adaptation may occur in longer space flights.

Simultaneous Analysis of Losartan Potassium, Amlodipine Besylate, and Hydrochlorothiazide in Bulk and in Tablets by High-Performance Thin Layer Chromatography with UV-Absorption Densitometry

PubMed Central

Santhana Lakshmi, Karunanidhi; Lakshmi, Sivasubramanian

2012-01-01

A Simple high-performance thin layer chromatography (HPTLC) method for separation and quantitative analysis of losartan potassium, amlodipine, and hydrochlorothiazide in bulk and in pharmaceutical formulations has been established and validated. After extraction with methanol, sample and standard solutions were applied to silica gel plates and developed with chloroform : methanol : acetone : formic acid 7.5 : 1.3 : 0.5 : 0.03 (v/v/v/v) as mobile phase. Zones were scanned densitometrically at 254 nm. The R f values of amlodipine besylate, hydrochlorothiazide, and losartan potassium were 0.35, 0.57, and 0.74, respectively. Calibration plots were linear in the ranges 500–3000 ng per spot for losartan potassium, amlodipine and hydrochlorothiazide, the correlation coefficients, r, were 0.998, 0.998, and 0.999, respectively. The suitability of this method for quantitative determination of these compounds was by validation in accordance with the requirements of pharmaceutical regulatory standards. The method can be used for routine analysis of these drugs in bulk and in formulation. PMID:22567550

Simultaneous determination of atenolol and amlodipine in tablets by high-performance thin-layer chromatography.

PubMed

Argekar, A P; Powar, S G

2000-01-01

A new simple, precise, rapid and selective high-performance thin-layer chromatographic (HPTLC) method has been developed for the simultaneous determination of atenolol (ATL) and amlodipine (AMLO) in tablets, using methylene chloride:methanol:ammonia solution (25% NH3) (8.8:1.3:0.1; v/v) as the mobile phase and Merck HPTLC plates (0.2 mm thickness) precoated with 60F254 silica gel on aluminium sheet as the stationary phase. Detection was carried out densitometrically using a UV detector at 230 nm. The retention factors of ATL and AMLO were 0.33 and 0.75, respectively. Calibration curves were linear in the range 10-500 microg ml(-1) for both. Assays of ATL and AMLO were 49.87 mg per tablet (relative standard deviation (R.S.D.), 1.3%) and 4.90 mg per tablet (R.S.D., 1.38%) for brand I, and 49.27 mg per tablet (R.S.D., 1.12%) and 4.98 mg per tablet (R.S.D., 1.42%) for brand II, respectively. The percentage recoveries for ATL and AMLO for brands I and II were 99.06 and 99.30%, and 99.27 and 99.15%, respectively.

[Morphological characteristic of erythrocytes in experimental hypervitaminosis A].

PubMed

Minashkina, T A

2011-01-01

This investigation was aimed at the analysis of the shape and morpho-densitometric parameters of the erythrocytes in rats with experimental hypervitaminosis A. Male Wistar rats received 0.64 mg/g (1167 IU/g) of retinol palmitate (RP) in oil solution orally for 11 consecutive days. Rats fed oil alone and intact animals were used as control groups. At days 5 and 6 of the experiment, the first manifestations of hypervitaminosis A were observed (body mass loss, localized erythema and hemorrhages). In contrast to control groups, in rats with hypervitaminosis A, the area of erythrocyte cytoplasm decreased gradually in response to RP administration. Discocyte/spherocyte/stomatocyte ratio also changed dynamically: the proportion of discocytes progressively decreased, while the amount of spherocytes and stomatocytes increased. These results show that excess of the vitamin A alters the erythrocyte membrane structure. Integral optical density of erythrocyte cytoplasm in RP-treated rats as well as in oil-fed rats was lower than in intact animals. This may be an indirect evidence of the fall in erythrocyte hemoglobin content. The changes observed in erythrocytes of RP-treated rats may serve as an additional criterion for evaluation of hypervitaminosis A severity.

Seed viability detection using computerized false-color radiographic image enhancement

NASA Technical Reports Server (NTRS)

Vozzo, J. A.; Marko, Michael

1994-01-01

Seed radiographs are divided into density zones which are related to seed germination. The seeds which germinate have densities relating to false-color red. In turn, a seed sorter may be designed which rejects those seeds not having sufficient red to activate a gate along a moving belt containing the seed source. This results in separating only seeds with the preselected densities representing biological viability lending to germination. These selected seeds demand a higher market value. Actual false-coloring isn't required for a computer to distinguish the significant gray-zone range. This range can be predetermined and screened without the necessity of red imaging. Applying false-color enhancement is a means of emphasizing differences in densities of gray within any subject from photographic, radiographic, or video imaging. Within the 0-255 range of gray levels, colors can be assigned to any single level or group of gray levels. Densitometric values then become easily recognized colors which relate to the image density. Choosing a color to identify any given density allows separation by morphology or composition (form or function). Additionally, relative areas of each color are readily available for determining distribution of that density by comparison with other densities within the image.

Development and validation of a HPTLC method for simultaneous estimation of lornoxicam and thiocolchicoside in combined dosage form

PubMed Central

Sahoo, Madhusmita; Syal, Pratima; Hable, Asawaree A.; Raut, Rahul P.; Choudhari, Vishnu P.; Kuchekar, Bhanudas S.

2011-01-01

Aim: To develop a simple, precise, rapid and accurate HPTLC method for the simultaneous estimation of Lornoxicam (LOR) and Thiocolchicoside (THIO) in bulk and pharmaceutical dosage forms. Materials and Methods: The separation of the active compounds from pharmaceutical dosage form was carried out using methanol:chloroform:water (9.6:0.2:0.2 v/v/v) as the mobile phase and no immiscibility issues were found. The densitometric scanning was carried out at 377 nm. The method was validated for linearity, accuracy, precision, LOD (Limit of Detection), LOQ (Limit of Quantification), robustness and specificity. Results: The Rf values (±SD) were found to be 0.84 ± 0.05 for LOR and 0.58 ± 0.05 for THIO. Linearity was obtained in the range of 60–360 ng/band for LOR and 30–180 ng/band for THIO with correlation coefficients r2 = 0.998 and 0.999, respectively. The percentage recovery for both the analytes was in the range of 98.7–101.2 %. Conclusion: The proposed method was optimized and validated as per the ICH guidelines. PMID:23781452

The application of LANDSAT-1 imagery for monitoring strip mines in the new river watershed in northeast Tennessee, part 2

NASA Technical Reports Server (NTRS)

Shahrokhi, F. (Principal Investigator); Sharber, L. A.

1977-01-01

The author has identified the following significant results. LANDSAT imagery and supplementary aircraft photography of the New River drainage basin were subjected to a multilevel analysis using conventional photointerpretation methods, densitometric techniques, multispectral analysis, and statistical tests to determine the accuracy of LANDSAT-1 imagery for measuring strip mines of common size. The LANDSAT areas were compared with low altitude measurements. The average accuracy over all the mined land sample areas mapped from LANDSAT-1 was 90%. The discrimination of strip mine subcategories is somewhat limited on LANDSAT imagery. A mine site, whether active or inactive, can be inferred by lack of vegetation, by shape, or image texture. Mine ponds are difficult or impossible to detect because of their small size and turbidity. Unless bordered and contrasted with vegetation, haulage roads are impossible to delineate. Preparation plants and refuge areas are not detectable. Density slicing of LANDSAT band 7 proved most useful in the detection of reclamation progress within the mined areas. For most state requirements for year-round monitoring of surface mined land, LANDSAT is of limited value. However, for periodic updating of regional surface maps, LANDSAT may provide sufficient accuracies for some users.

Susceptibility to kinetotic Behaviour during Parabolic Aircraft Flights and otolithic Calcium Incorporation in Fish

NASA Astrophysics Data System (ADS)

Forster, A.; Anken, R.; Hilbig, R.

According to an earlier concept, otolith (or statolith) asymmetry is the cause for susceptibility to kinetoses (e.g., human static space sickness). Indeed, we could recently show that fish showing a kinetotic behaviour after development at hypergravity had incorporated significantly more otolithic calcium (and had an higher otolith asymmetry concerning calcium incorporation) as had normally swimming hyper-g specimens. In order to determine whether a (predispositioned) high asymmetry of otolithic calcium incorporation may also be the cause for kinetosis susceptibility in the microgravity environment (to be achived during parabolic aircraft flights, PFs), larval cichlid fish (Oreochromis mossambicus) were (prior to the PFs) maintained in aquarium water containing alizarin-complexone (AC), a fluorescent calcium tracer. Subsequently, the behaviour of the animals during the microgravity phases of the PF experiment was qualitatively assessed and the specimens were seperated into normally and kinetotically swimming individuals (the latter performed spinning movements). Finally, otolithic AC (and thus calzium) incorporation was densitometrically determined in the otoliths and correlated with the animals' behavior. The respective data will be communicated at the meeting. Acknowledgement: This work was financially supported by the German Aerospace Center (DLR) (FKZ: 50 WB 9997).

Simultaneous quantification of stevioside and rebaudioside A in different stevia samples collected from the Indian subcontinent

PubMed Central

Chester, Karishma; Tamboli, Ennus T.; Singh, Mhaveer; Ahmad, Sayeed

2012-01-01

Background: A high performance thin layer chromatographic (HPTLC) method was developed for simultaneous estimation of stevioside and rebaudioside A in Stevia rebaudiana samples collected from different regions of Indian subcontinent. Materials and Methods: The separation was achieved by using acetone: ethyl acetate: water (5:4:1, v/v/v) as the solvent system on precoated silica gel 60 F254 TLC plates. The densitometric quantification of stevia glycosides was carried out at wavelength 360 nm in absorption mode after spraying with anisaldehyde sulphuric acid as detecting reagent. Results: The well resolved peaks for stevioside and rebaudioside A were observed at Rf values 0.31± 0.02 and 0.21± 0.02 respectively. The calibration curves were found linear with a wide range of concentration 100 - 2000 ng spot-1 with good correlation coefficient 0.996 and 0.991 for stevioside and rebaudioside A, respectively. Conclusions: The proposed method was validated as per the ICH (International Conferences on Harmonization) guidelines and found simple, sensitive, economic, reproducible, robust and accurate for quantitative analysis of stevia glycosides, which can be applied for quality control of stevia as well as to check. PMID:23248559

[Osteoporosis in all young daughters of a mother with multiple osteoporotic fractures. A case of familial osteoporosis].

PubMed

Parisi, M S; Díaz, A G; Oliveri, M B; Di Gregorio, S; Mautalen, C A

2001-01-01

We herein describe a family whose female members are all osteoporotic: a postmenopausal mother and her three premenopausal daughters. The mother aged 60 presented axial and peripheral fractures, and very low bone mineral density (BMD). She reported that her grandmother had suffered a hip fracture. The eldest daughter aged 30 suffered multiple vertebral fractures during pregnancy and lactation associated with very low BMD. In view of these observations, the other two daughters aged 29 and 27 years respectively were evaluated. BMD was found to be severely diminished according to densitometric values for osteoporosis established by WHO, but they had no history of bone fractures. Probably the strong genetic component in bone mass is responsible for the severely diminished BMD observed in all the women in this family, as well as the occurrence of bone fractures in two of them. To our knowledge, there are no similar reports in the literature. Our results evidence the importance of evaluating bone mass in the offspring of an individual presenting severe osteoporosis, in order to detect family members with low bone mass and at high risk of developing bone fractures.

Spinal Bone Texture Assessed by Trabecular Bone Score in Adolescent Girls With Anorexia Nervosa

PubMed Central

Donaldson, Abigail A.; Feldman, Henry A.; O'Donnell, Jennifer M.; Gopalakrishnan, Geetha

2015-01-01

Context: Trabecular bone score (TBS) is a bone assessment tool that offers information beyond that afforded by dual-energy x-ray absorptiometry (DXA) bone mineral density (BMD) measurements. Adolescents with anorexia nervosa (AN) are known to exhibit compromised bone density and skeletal strength. Objectives: This study aimed to determine TBS among adolescents with AN and evaluate the correlation with anthropometric, clinical and densitometric variables. Design: Areal BMD spinal measures were analyzed for TBS. Findings were compared with clinical (height, weight, body mass index [BMI], age, pubertal development, 25-hydroxyvitamin D) and self-reported data (illness duration, amenorrhea, exercise, fracture, family history of osteoporosis, and antidepressant use), and BMD measures by DXA and peripheral quantitative computed tomography (pQCT). Setting and Participants: This was an urban adolescent program consisting of 57 females with AN, age 11–18 y. Interventions: Interventions included DXA (absolute BMD and Z-score), pQCT (volumetric BMD [vBMD] and stress-strain index [SSI]), laboratory evaluation, and questionnaire administration. Main Outcome Measures: Main outcome measures included TBS, areal and vBMD, SSI, fracture history, disease duration. Results: The TBS of six participants (11%) showed degraded and 19 (33%) partially degraded microarchitecture. Spinal TBS was correlated (P < .05) with age, height, weight, BMI, pubertal stage, BMD, and body composition by DXA, and BMD and SSI by pQCT. TBS was not correlated with disease duration, fracture, vitamin D status, race, or ethnicity, and self-reported health data. Conclusions: TBS showed evidence of degraded microarchitecture in over 40% of this study sample, and strongly correlated with anthropometric data and measures of BMD and skeletal strength. TBS is a novel tool that captures another dimension of bone health in adolescents with AN. PMID:26108094

Effects of combined maternal administration with alpha-ketoglutarate (AKG) and β-hydroxy-β-methylbutyrate (HMB) on prenatal programming of skeletal properties in the offspring.

PubMed

Tatara, Marcin R; Krupski, Witold; Tymczyna, Barbara; Studziński, Tadeusz

2012-05-11

Nutritional manipulations during fetal growth may induce long-term metabolic effects in postnatal life. The aim of the study was to test whether combined treatment of pregnant sows with alpha-ketoglutarate and β-hydroxy-β-methylbutyrate induces additive long-term effects on skeletal system properties in the offspring. The study was performed on 290 pigs obtained from 24 sows divided into 4 equal groups and subjected to experimental treatment during two weeks before delivery. The first group consisted of control sows, while the second group received alpha-ketoglutarate. The third group was treated with β-hydroxy-β-methylbutyrate and the fourth group underwent combined administration of alpha-ketoglutarate and β-hydroxy-β-methylbutyrate. Piglets obtained from sows were reared until slaughter age to perform morphometric, densitometric and mechanical analyses of femur. Serum evaluations of growth hormone, insulin-like growth factor-1, bone-specific alkaline phosphatase and osteocalcin were performed in newborns and 90-day old piglets; additionally, plasma amino acid concentration was measured in newborns. Maternal treatment with alpha-ketoglutarate and β-hydroxy-β-methylbutyrate significantly reduced fattening time and increased birth body weight, daily body weight gain, bone weight, volumetric bone mineral density, geometrical parameters and mechanical endurance of femur. These effects were associated with increased serum concentrations of growth hormone, insulin-like growth factor-1, bone-specific alkaline phosphatase and osteocalcin. Furthermore, alpha-ketoglutarate and β-hydroxy-β-methylbutyrate administered solely or in combination significantly increased plasma level of 19 amino acids. Hormonal and amino acid evaluations in pigs indicate additive effects of AKG and HMB on systemic growth and development; however, determination of bone properties has not shown such phenomenon.

Effects of combined maternal administration with alpha-ketoglutarate (AKG) and β-hydroxy-β-methylbutyrate (HMB) on prenatal programming of skeletal properties in the offspring

PubMed Central

2012-01-01

Background Nutritional manipulations during fetal growth may induce long-term metabolic effects in postnatal life. The aim of the study was to test whether combined treatment of pregnant sows with alpha-ketoglutarate and β-hydroxy-β-methylbutyrate induces additive long-term effects on skeletal system properties in the offspring. Methods The study was performed on 290 pigs obtained from 24 sows divided into 4 equal groups and subjected to experimental treatment during two weeks before delivery. The first group consisted of control sows, while the second group received alpha-ketoglutarate. The third group was treated with β-hydroxy-β-methylbutyrate and the fourth group underwent combined administration of alpha-ketoglutarate and β-hydroxy-β-methylbutyrate. Piglets obtained from sows were reared until slaughter age to perform morphometric, densitometric and mechanical analyses of femur. Serum evaluations of growth hormone, insulin-like growth factor-1, bone-specific alkaline phosphatase and osteocalcin were performed in newborns and 90-day old piglets; additionally, plasma amino acid concentration was measured in newborns. Results Maternal treatment with alpha-ketoglutarate and β-hydroxy-β-methylbutyrate significantly reduced fattening time and increased birth body weight, daily body weight gain, bone weight, volumetric bone mineral density, geometrical parameters and mechanical endurance of femur. These effects were associated with increased serum concentrations of growth hormone, insulin-like growth factor-1, bone-specific alkaline phosphatase and osteocalcin. Furthermore, alpha-ketoglutarate and β-hydroxy-β-methylbutyrate administered solely or in combination significantly increased plasma level of 19 amino acids. Conclusions Hormonal and amino acid evaluations in pigs indicate additive effects of AKG and HMB on systemic growth and development; however, determination of bone properties has not shown such phenomenon. PMID:22578071

GABAergic system impairment in the hippocampus and superior temporal gyrus of patients with paranoid schizophrenia: A post-mortem study.

PubMed

Steiner, Johann; Brisch, Ralf; Schiltz, Kolja; Dobrowolny, Henrik; Mawrin, Christian; Krzyżanowska, Marta; Bernstein, Hans-Gert; Jankowski, Zbigniew; Braun, Katharina; Schmitt, Andrea; Bogerts, Bernhard; Gos, Tomasz

2016-11-01

Glutamic acid decarboxylase (GAD) is a key enzyme in GABA synthesis and alterations in GABAergic neurotransmission related to glial abnormalities are thought to play a crucial role in the pathophysiology of schizophrenia. This study aimed to identify potential differences regarding the neuropil expression of GAD between paranoid and residual schizophrenia. GAD65/67 immunostained histological sections were evaluated by quantitative densitometric analysis of GAD-immunoreactive (ir) neuropil. Regions of interest were the hippocampal formation (CA1 field and dentate gyrus [DG]), superior temporal gyrus (STG), and laterodorsal thalamic nucleus (LD). Data from 16 post-mortem schizophrenia patient samples (10 paranoid and 6 residual schizophrenia cases) were compared with those from 16 matched controls. Overall, schizophrenia patients showed a lower GAD-ir neuropil density (P=0.014), particularly in the right CA1 (P=0.033). However, the diagnostic subgroups differed significantly (P<0.001), mainly because of lower right CA1 GAD-ir neuropil density in paranoid versus residual patients (P=0.036) and controls (P<0.003). Significant GAD-ir neuropil reduction was also detected in the right STG layer V of paranoid versus residual schizophrenia cases (P=0.042). GAD-ir neuropil density correlated positively with antipsychotic dosage, particularly in CA1 (right: r=0.850, P=0.004; left: r=0.800, P=0.010). Our finding of decreased relative density of GAD-ir neuropil suggests hypofunction of the GABAergic system, particularly in hippocampal CA1 field and STG layer V of patients with paranoid schizophrenia. The finding that antipsychotic medication seems to counterbalance GABAergic hypofunction in schizophrenia patients suggests the possibility of exploring new treatment avenues which target this system. Copyright © 2016 Elsevier B.V. All rights reserved.

Analysis of the influence of the T393C polymorphism of the GNAS gene on the clinical expression of primary hyperparathyroidism.

PubMed

Piedra, María; Berja, Ana; Ramos, Laura; García-Unzueta, María Teresa; Morán, Jesús Manuel; Ruiz, David; Amado, José Antonio

2017-12-01

The receptor of parathyroid hormone and parathyroid hormone-related-protein (PTH/PTHrp) is located in the cell membrane of target tissues - kidney and osteoblasts. It is a G protein-coupled-receptor whose G s α subunit is encoded by the GNAS gene. Our aim was to study whether the single nucleotide polymorphism (SNP) T393C of the GNAS gene is associated with renal stones, bone mineral density (BMD), or bone remodelling markers in primary hyperparathyroidism (PHPT). An analysis was made of clinical and biochemical parameters and densitometric values in three areas and their relationship with the T393C SNP of the GNAS gene in 261 patients with primary hyperparathyroidism and in 328 healthy controls. Genotyping was performed using the Custom Taqman ® SNP Genotyping assay. The genotype frequencies of GNAS T/C 393 were similar in the control and PHPT groups. No association was found between genotypes and clinical expression of PHPT (renal stones and bone fractures). A nonstatistically significant trend was seen to lower BMD in the lumbar spine, femoral neck, and total hip in both PHPT and control C homozygote subjects. Genetic susceptibility to PHPT related to the GNAS T393C polymorphism or a major influence in its development and clinical expression were found. A C allele-related susceptibility to lower BMD in trabecular bone in both PHPT and control subjects is not sufficient to suggest a more severe clinical expression of PHPT. This trend may be considered as a basis for further studies with larger sample sizes and complementary functional evaluation. Copyright © 2017 SEEN y SED. Publicado por Elsevier España, S.L.U. All rights reserved.

Establishing Biomechanical Mechanisms in Mouse Models: Practical Guidelines for Systematically Evaluating Phenotypic Changes in the Diaphyses of Long Bones

PubMed Central

Jepsen, Karl J; Silva, Matthew J; Vashishth, Deepak; Guo, X Edward; van der Meulen, Marjolein CH

2016-01-01

Mice are widely used in studies of skeletal biology, and assessment of their bones by mechanical testing is a critical step when evaluating the functional effects of an experimental perturbation. For example, a gene knockout may target a pathway important in bone formation and result in a “low bone mass” phenotype. But how well does the skeleton bear functional loads; eg, how much do bones deform during loading and how resistant are bones to fracture? By systematic evaluation of bone morphological, densitometric, and mechanical properties, investigators can establish the “biomechanical mechanisms” whereby an experimental perturbation alters whole-bone mechanical function. The goal of this review is to clarify these biomechanical mechanisms and to make recommendations for systematically evaluating phenotypic changes in mouse bones, with a focus on long-bone diaphyses and cortical bone. Further, minimum reportable standards for testing conditions and outcome variables are suggested that will improve the comparison of data across studies. Basic biomechanical principles are reviewed, followed by a description of the cross-sectional morphological properties that best inform the net cellular effects of a given experimental perturbation and are most relevant to biomechanical function. Although morphology is critical, whole-bone mechanical properties can only be determined accurately by a mechanical test. The functional importance of stiffness, maximum load, postyield displacement, and work-to-fracture are reviewed. Because bone and body size are often strongly related, strategies to adjust whole-bone properties for body mass are detailed. Finally, a comprehensive framework is presented using real data, and several examples from the literature are reviewed to illustrate how to synthesize morphological, tissue-level, and whole-bone mechanical properties of mouse long bones. PMID:25917136

Hsp70 expression in thermally stressed Ostrea edulis, a commercially important oyster in Europe

PubMed Central

Piano, Annamaria; Asirelli, Christian; Caselli, Federico; Fabbri, Elena

2002-01-01

Synthesis of heat shock proteins (Hsps) in response to elevated temperatures and other denaturing agents is a common feature of prokaryotic and eukaryotic cells. The heat-induced expression of Hsp70 family members in the gills and mantle of Ostrea edulis, a highly valued fisheries resource inhabiting primarily estuarine environments, has been studied. O edulis is exposed to a variety of natural and anthropogenic stresses in the environment. Two isoforms of about 72 kDa and 77 kDa were constitutively present in unstressed organisms, reflecting the housekeeping function performed by these proteins under normal circumstances. Their expression in animals undergoing thermal stress was highly variable, and on the average, little change occurred under different experimental conditions. A third isoform of about 69 kDa was induced in both tissues after exposure to ≥32°C; its synthesis was detected within 4 hours of poststress recovery at 15°C, reaching the maximum expression after 24 hours in the gills and after 48 hours in the mantle and declining thereafter. Hsp69 expression was low at 38°C, a temperature lethal for about 50% of the individuals tested. Densitometric analysis of Western blots revealed that Hsp69 was mostly responsible for the significant heat-induced overexpression of Hsp70s in O edulis. Comparison with heat shock responses in tissues of Crassostrea gigas indicated a similar pattern of Hsp70 expression. In this organism, however, Hsp69 was induced after exposure to ≥38°C. We conclude that tissue expression of Hsp69 in O edulis, and possibly other bivalves, is an early sign of thermal stress; determining whether these changes also correlate with other major environmental stresses is the goal of ongoing studies. PMID:12482201

Development and validation of high-performance liquid chromatography and high-performance thin-layer chromatography methods for the quantification of khellin in Ammi visnaga seed

PubMed Central

Kamal, Abid; Khan, Washim; Ahmad, Sayeed; Ahmad, F. J.; Saleem, Kishwar

2015-01-01

Objective: The present study was used to design simple, accurate and sensitive reversed phase-high-performance liquid chromatography RP-HPLC and high-performance thin-layer chromatography (HPTLC) methods for the development of quantification of khellin present in the seeds of Ammi visnaga. Materials and Methods: RP-HPLC analysis was performed on a C18 column with methanol: Water (75: 25, v/v) as a mobile phase. The HPTLC method involved densitometric evaluation of khellin after resolving it on silica gel plate using ethyl acetate: Toluene: Formic acid (5.5:4.0:0.5, v/v/v) as a mobile phase. Results: The developed HPLC and HPTLC methods were validated for precision (interday, intraday and intersystem), robustness and accuracy, limit of detection and limit of quantification. The relationship between the concentration of standard solutions and the peak response was linear in both HPLC and HPTLC methods with the concentration range of 10–80 μg/mL in HPLC and 25–1,000 ng/spot in HPTLC for khellin. The % relative standard deviation values for method precision was found to be 0.63–1.97%, 0.62–2.05% in HPLC and HPTLC for khellin respectively. Accuracy of the method was checked by recovery studies conducted at three different concentration levels and the average percentage recovery was found to be 100.53% in HPLC and 100.08% in HPTLC for khellin. Conclusions: The developed HPLC and HPTLC methods for the quantification of khellin were found simple, precise, specific, sensitive and accurate which can be used for routine analysis and quality control of A. visnaga and several formulations containing it as an ingredient. PMID:26681890

Gene expression analysis in zebrafish embryos: a potential approach to predict effect concentrations in the fish early life stage test.

PubMed

Weil, Mirco; Scholz, Stefan; Zimmer, Michaela; Sacher, Frank; Duis, Karen

2009-09-01

Based on the hypothesis that analysis of gene expression could be used to predict chronic fish toxicity, the zebrafish (Danio rerio) embryo test (DarT), developed as a replacement method for the acute fish test, was expanded to a gene expression D. rerio embryo test (Gene-DarT). The effects of 14 substances on lethal and sublethal endpoints of the DarT and on expression of potential marker genes were investigated: the aryl hydrocarbon receptor 2, cytochrome P450 1A (cypla), heat shock protein 70, fizzy-related protein 1, the transcription factors v-maf musculoaponeurotic fibrosarcoma oncogene family protein g (avian) 1 and NF-E2-p45-related factor, and heme oxygenase 1 (hmox1). After exposure of zebrafish embryos for 48 h, differential gene expression was evaluated using reverse transcriptase-polymerase chain reaction, gel electrophoresis, and densitometric analysis of the gels. All tested compounds significantly affected the expression of at least one potential marker gene, with cyp1a and hmox1 being most sensitive. Lowest-observed-effect concentrations (LOECs) for gene expression were below concentrations resulting in 10% lethal effects in the DarT. For 10 (3,4- and 3,5-dichloroaniline, 1,4-dichlorobenzene, 2,4-dinitrophenol, atrazine, parathion-ethyl, chlorotoluron, genistein, 4-nitroquinoline-1-oxide, and cadmium) out of the 14 tested substances, LOEC values derived with the Gene-DarT differ by a factor of less than 10 from LOEC values of fish early life stage tests with zebrafish. For pentachloroaniline and pentachlorobenzene, the Gene-DarT showed a 23- and 153-fold higher sensitivity, respectively, while for lindane, it showed a 13-fold lower sensitivity. For ivermectin, the Gene-DarT was by a factor of more than 1,000 less sensitive than the acute fish test. The results of the present study indicate that gene expression analysis in zebrafish embryos could principally be used to predict effect concentrations in the fish early life stage test.

The peripheral quantitative computed tomographic and densitometric analysis of skeletal tissue in male Wistar rats after chromium sulfate treatment.

PubMed

Bieńko, Marek; Radzki, Radosław Piotr; Wolski, Dariusz

2017-09-21

This study evaluates the effects of three different doses of chromium sulphate on bone density and the tomographic parameters of skeletal tissue of rats. The experiment was performed on 40 male Wistar rats which received, by gavage, during 90 days, a chromium sulphate in either a daily dose of 400, 600 or 800 µg/kg BW. At the end of experiment, the rats were scanned using the densitometry method (DXA) to determine the bone mineral density, bone mineral content of total skeleton and vertebral column (L2-L4) and parameters of body composition (Lean Mass and Fat Mass). The isolated femora were scanned using peripheral a quantitative computed tomography method (pQCT) for a separate analysis of the trabecular and cortical bone tissue. The ultimate strength, work to ultimate and the Young modulus of femora was also investigated by the three-point bending test. The negative impact of chromium was observed in relation to bone tissue. All doses significantly decreased total skeleton density and mineral content, and also had impact upon the isolated femora and vertebral column. Trabecular volumetric bone mineral density and trabecular bone mineral content measured by pQCT in distal femur metaphysis were significantly lower in the experimental groups than in the control. Higher doses of chromium also significantly decreased values of ultimate strength and Young modulus in the investigated femora. The results of the experiment demonstrate that chromium sulphate is dose dependent, and exerts a disadvantageous effect on the skeleton, as it decreases bone density and resistance.

PCR-denaturing gradient gel electrophoresis analysis of microbial community in soy-daddawa, a Nigerian fermented soybean (Glycine max (L.) Merr.) condiment.

PubMed

Ezeokoli, Obinna T; Gupta, Arvind K; Mienie, Charlotte; Popoola, Temitope O S; Bezuidenhout, Cornelius C

2016-03-02

Soy-daddawa, a fermented soybean (Glycine max (L.) Merr.) condiment, plays a significant role in the culinary practice of West Africa. It is essential to understand the microbial community of soy-daddawa for a successful starter culture application. This study investigated the microbial community structure of soy-daddawa samples collected from Nigerian markets, by PCR-denaturing gradient gel electrophoresis (DGGE) targeting the V3-V5 region of the 16S rRNA gene of bacteria and internal transcribed spacer 2 (ITS2) region of fungi. Six bacterial and 16 fungal (nine yeasts and seven molds) operational taxonomic units (OTUs)/species were obtained at 97% sequence similarity. Taxonomic assignments revealed that bacterial OTUs belonged to the phyla Firmicutes and Actinobacteria, and included species from the genera Atopostipes, Bacillus, Brevibacterium and Nosocomiicoccus. Densitometric analysis of DGGE image/bands revealed that Bacillus spp. were the dominant OTU/species in terms of population numbers. Fungal OTUs belonged to the phyla Ascomycota and Zygomycota, and included species from the genera, Alternaria, Aspergillus, Candida, Cladosporium, Dokmaia, Issatchenkia, Kodamaea, Lecythophora, Phoma, Pichia, Rhizopus, Saccharomyces and Starmerella. The majority of fungal species have not been previously reported in soy-daddawa. Potential opportunistic human pathogens such as Atopostipes suicloacalis, Candida rugosa, Candida tropicalis, and Kodamaea ohmeri were detected. Variation in soy-daddawa microbial communities amongst samples and presence of potential opportunistic pathogens emphasises the need for starter culture employment and good handling practices in soy-daddawa processing. Copyright © 2016 Elsevier B.V. All rights reserved.

Anti-LRP/LR Specific Antibody IgG1-iS18 Impedes Adhesion and Invasion of Liver Cancer Cells

PubMed Central

Chetty, Carryn; Khumalo, Thandokuhle; Da Costa Dias, Bianca; Reusch, Uwe; Knackmuss, Stefan; Little, Melvyn; Weiss, Stefan F. T.

2014-01-01

Two key events, namely adhesion and invasion, are pivotal to the occurrence of metastasis. Importantly, the 37 kDa/67 kDa laminin receptor (LRP/LR) has been implicated in enhancing these two events thus facilitating cancer progression. In the current study, the role of LRP/LR in the adhesion and invasion of liver cancer (HUH-7) and leukaemia (K562) cells was investigated. Flow cytometry revealed that the HUH-7 cells displayed significantly higher cell surface LRP/LR levels compared to the poorly-invasive breast cancer (MCF-7) control cells, whilst the K562 cells displayed significantly lower cell surface LRP/LR levels in comparison to the MCF-7 control cells. However, Western blotting and densitometric analysis revealed that all three tumorigenic cell lines did not differ significantly with regards to total LRP/LR levels. Furthermore, treatment of liver cancer cells with anti-LRP/LR specific antibody IgG1-iS18 (0.2 mg/ml) significantly reduced the adhesive potential of cells to laminin-1 and the invasive potential of cells through the ECM-like Matrigel, whilst leukaemia cells showed no significant differences in both instances. Additionally, Pearson's correlation coefficients suggested direct proportionality between cell surface LRP/LR levels and the adhesive and invasive potential of liver cancer and leukaemia cells. These findings suggest the potential use of anti-LRP/LR specific antibody IgG1-iS18 as an alternative therapeutic tool for metastatic liver cancer through impediment of the LRP/LR- laminin-1 interaction. PMID:24798101

Measurement of percent oxyhemoglobin by optical densitometry in perfluorocarbon supplemented blood.

PubMed

Murrah, C P; Agnihotri, A K; Spruell, R D; Clymer-Hawk, J J; Ferguson, E R; Wiley, J H; Holman, W L

1996-01-01

A new generation of perfluorocarbon emulsions is being clinically evaluated as erythrocyte substitutes. However, the effect of perfluorocarbon emulsions on optical densitometric measurements of percent oxyhemoglobin (%O2Hb) has not been fully characterized. The purpose of this study is to quantify the effect of blood perfluorochemical concentration and hematocrit (Hct) on %O2Hb measurements. The authors hypothesize that perfluorocarbon emulsions affect the accuracy of %O2Hb measurements, and that the effect of perfluorochemical concentration and Hct on these measurements can be mathematically described. Porcine blood was used in this experiment. Blood with a Hct of 18% or 9% was mixed with a perfluorocarbon emulsion (Oxygent [AF0142]; Alliance Pharmaceutical Corporation, San Diego, CA). The concentrations tested were 0 g (Group I; n = 69 measurements for a Hct of 18%, and n = 35 measurements for a Hct of 9%), 0.73 g (Group II; n = 47 at 18%, n = 33 at 9%), 1.45 g (Group III; n = 46 at 18%, n = 30 at 9%), and 2.90 g (Group IV; n = 45 at 18%, n = 31 at 9%) of perfluorochemical per deciliter of blood (g PFC/dl). A tonometer was used to establish a range of oxygen tensions within each group while maintaining physiologic pH and PCO2. Error in %O2Hb measurements increases with higher perfluorochemical concentrations and lower Hct values. These errors in %O2Hb measurements are predictable; as such, an equation for correcting %O2Hb measurements in perfluorocarbon supplemented blood can be generated.

Radiographic morphometry and densitometry predict strength of cadaveric proximal humeri more reliably than age and DXA scan density.

PubMed

Skedros, John G; Knight, Alex N; Pitts, Todd C; O'Rourke, Peter J; Burkhead, Wayne Z

2016-02-01

Methods are needed for identifying poorer quality cadaver proximal humeri to ensure that they are not disproportionately segregated into experimental groups for fracture studies. We hypothesized that measurements made from radiographs of cadaveric proximal humeri are stronger predictors of fracture strength than chronological age or bone density values derived from dual-energy x-ray absorptiometry (DXA) scans. Thirty-three proximal humeri (range: 39-78 years) were analyzed for: (1) bone mineral density (BMD, g/cm(2)) using DXA, (2) bulk density (g/cm(3)) using DXA and volume displacement, (3) regional bone density in millimeters of aluminum (mmAl) using radiographs, and (4) regional mean (medial+lateral) cortical thickness and cortical index (CI) using radiographs. The bones were then fractured simulating a fall. Strongest correlations with ultimate fracture load (UFL) were: mean cortical thickness at two diaphyseal locations (r = 0.71; p < 0.001), and mean mmAl in the humeral head (r = 0.70; p < 0.001). Weaker correlations were found between UFL and DXA-BMD (r = 0.60), bulk density (r = 0.43), CI (r = 0.61), and age (r = -0.65) (p values <0.01). Analyses between UFL and the product of any two characteristics showed six combinations with r-values >0.80, but none included DXA-derived density, CI, or age. Radiographic morphometric and densitometric measurements from radiographs are therefore stronger predictors of UFL than age, CI, or DXA-derived density measurements. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

Relationship between pre-sarcopenia, sarcopenia and bone mineral density in elderly men.

PubMed

Pereira, Fernando Borges; Leite, André Ferreira; de Paula, Ana Patrícia

2015-02-01

Analyze the influence of sarcopenia in bone health of elderly men. This cross-sectional study evaluated 198 men aged over 60 years. Body composition was measured by dual energy X-ray absorptiometry. The BMD was measured at the femoral neck, total hip, lumbar spine and 33% radius. The diagnosis of abnormal BMD was defined for men who presented densitometric diagnosis of osteopenia or osteoporosis defined by T-score of femoral neck, total hip and lumbar spine. The pre-sarcopenia and sarcopenia were defined according to the European Working Group on Sarcopenia in Older People. The group diagnosed with normal BMD, compared to the group of abnormal BMD, have significantly higher body weight, body mass index, grip strength, lean mass, fat mass, and relative appendicular skeletal muscle mass (RASM). However, after multiple linear regression analysis, we found that only the RASM, lean mass, and handgrip strength in the dominant hand influenced the variability of the BMD after adjustment for age and weight. Regression analyzes showed a positive association between greater appendicular lean mass and a smaller number of elderly patients with abnormal BMD diagnostic. The regression analyzes showed that elderly men diagnosed with pre-sarcopenia and sarcopenia had more abnormal BMD than non-sarcopenic elderly men. We concluded that pre-sarcopenia and sarcopenia were associated with abnormal BMD. The lean mass, compared to fat mass, has a greater positive influence on the BMD of elderly men. This result suggests the importance of the increase in lean mass for the bone health of elderly men.

The peak bone mass concept: is it still relevant?

PubMed

Schönau, Eckhard

2004-08-01

The peak bone mass concept implies that optimal skeletal development during childhood and adolescence will prevent fractures in late adulthood. This concept is based on the observation that areal bone density increases with growth during childhood, is highest around 20 years of age and declines thereafter. However, it is now clear that strong bones in the youngster do not necessarily lead to a fracture-free old age. In the recent bone densitometric literature, the terms bone mass and bone density are typically used synonymously. In physics, density has been defined as the mass of a body divided by its volume. In clinical practice and science, "bone density" usually has a different meaning-the degree to which a radiation beam is attenuated by a bone, as judged from a two-dimensional projection image (areal bone density). The attenuation of a radiation beam does not only depend on physical density, but also on bone size. A small bone therefore has a lower areal bone density than a larger bone, even if the physical density is the same. Consequently, a low areal bone density value can simply reflect the small size of an otherwise normal bone. At present, bone mass analysis is very useful for epidemiological studies on factors that may have an impact on bone development. There is an ongoing discussion about whether the World Health Organization (WHO) definition of osteoporosis is over-simplistic and requires upgrading to include indices representing the distribution of bone and mineral (bone strength indices). The following suggestions and recommendations outline a new concept: bone mass should not be related to age. There is now more and more evidence that bone mass should be related to bone size or muscle function. Thus analyzed, there is no such entity as a "peak bone mass". Many studies are currently under way to evaluate whether these novel approaches increase sensitivity and specificity of fracture prediction in an individual. Furthermore, the focus of many bone researchers is shifting away from bone mass to bone geometry or bone strength. Bone mass is one surrogate marker of bone strength. Widely available techniques for measurement of bone mass, such as dual-energy X-ray absorptiometry, radiogrammetry, and computed tomography, can also be used to measure variables of bone geometry such as cortical thickness, cortical area, and moment of inertia.

Rapid and cost-effective determination of acrylamide in coffee by planar chromatography and fluorescence detection after derivatization with dansulfinic acid.

PubMed

Alpmann, Alexander; Morlock, Gertrud

2009-01-01

A new method has been developed for the determination of acrylamide in ground coffee by planar chromatography using prechromatographic in situ derivatization with dansulfinic acid. After pressurized fluid extraction of acrylamide from the coffee samples, the extracts were passed through activated carbon and concentrated. These extracts were applied onto a silica gel 60 HPTLC plate and oversprayed with dansulfinic acid. By heating the plate, acrylamide was derivatized into the fluorescent product dansylpropanamide. The chromatographic separation with ethyl acetate-tert.-butyl methyl ether (8 + 2, v/v) mobile phase was followed by densitometric quantification at 254/>400 nm using a 4 point calibration via the standard addition method over the whole system for which acrylamide was added at different concentrations at the beginning of the extraction process. The method was validated for commercial coffee. The linearity over the whole procedure showed determination coefficients between 0.9995 and 0.9825 (n = 6). Limit of quantitation at a signal-to-noise ratio of 10 was determined to be 48 microg/kg. The within-run precision (relative standard deviation, n = 6) of the chromatographic method was 3%. Commercial coffee samples analyzed showed acrylamide contents between 52 and 191 microg/kg, which was in correlation with amounts reported in previous publications.

HPTLC Method for the Determination of Paracetamol, Pseudoephedrine and Loratidine in Tablets and Human Plasma

PubMed Central

Farid, Nehal Fayek; Abdelaleem, Eglal A.

2016-01-01

A sensitive, accurate and selective high performance thin layer chromatography (HPTLC) method was developed and validated for the simultaneous determination of paracetamol (PAR), its toxic impurity 4-aminophenol (4-AP), pseudoephedrine HCl (PSH) and loratidine (LOR). The proposed chromatographic method has been developed using HPTLC aluminum plates precoated with silica gel 60 F254 using acetone–hexane–ammonia (4:5:0.1, by volume) as a developing system followed by densitometric measurement at 254 nm for PAR, 4-AP and LOR, while PSH was scanned at 208 nm. System suitability testing parameters were calculated to ascertain the quality performance of the developed chromatographic method. The method was validated with respect to USP guidelines regarding accuracy, precision and specificity. The method was successfully applied for the determination of PAR, PSH and LOR in ATSHI® tablets. The three drugs were also determined in plasma by applying the proposed method in the ranges of 0.5–6 µg/band, 1.6–12 µg/band and 0.4–2 µg/band for PAR, PSH and LOR, respectively. The results obtained by the proposed method were compared with those obtained by a reported HPLC method, and there was no significance difference between both methods regarding accuracy and precision. PMID:26762956

HPTLC detection of altitudinal variation of the potential antivenin stigmasterol in different populations of the tropical ethnic antidote Rauvolfia serpentina.

PubMed

Dey, Abhijit; Pandey, Devendra Kumar

2014-09-01

To determine the altitudinal variation of stigmasterol, a potential antivenin, in roots from seven populations of Rauvolfia serpentina (L). Benth. ex Kurz. (Apocynaceae) (R. serpentina), an important herb found in Indian subcontinent which has long been used in the treatment of snakebite, blood pressure and schizophrenia. Altitudinal variation of stigmasterol content in R. serpentina roots was analyzed by high performance thin layer chromatography. Chromatography was performed on silica gel 60 F254 thin layer chromatography plates with benzene-acetone 86:14 (v/v) as mobile phase. Densitometric analysis was done at λ=366 nm after derivatization with vanillin-10% (v/v) sulphuric acid alcohol reagent. The method was validated for precision and recovery. The present experiment demonstrates a simple, rapid, precise and sensitive high performance thin layer chromatography protocol for qualitative and quantitative determination of stigmasterol from different populations of R. serpentina. Results demonstrated that in root samples stigmasterol was present at Rf value of 0.44. This investigation demonstrates that stigmasterol content in R. serpentina roots varies in different altitudes. Popular ethnomedicinal use of this herb against snakebite may be contributed by the occurrence of stigmasterol in its roots. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Osteoporosis diagnosis in men: the T-score controversy revisited.

PubMed

Binkley, Neil; Adler, Robert; Bilezikian, John P

2014-12-01

Osteoporosis becomes common with aging in both sexes, but is often ignored in men. The 2013 International Society for Clinical Densitometry consensus conference endorsed a Caucasian female referent database for T-score calculation in men. This recommendation has generated controversy and concern. Accumulating data indicate that at the same DXA-measured body mineral density (BMD) (g/cm(2)), men and women are at approximately the same fracture risk. With this point in mind, using the same database to derive the T-score in men and women is reasonable. As a result, a greater proportion of men who sustain a fragility fracture will have T-scores that are higher than they would if a male database were used; in fact, many men will fracture at T-scores that are "normal." This highlights the importance of diagnosing osteoporosis not just by T-score, but also by the presence of fragility fracture and/or by estimations of fracture risk as generated by tools such as the FRAX calculator. The practical consequences of this change in densitometric definition of osteoporosis in men should be monitored, including the proportion of men at risk identified and treated as well as defining the response to treatment in those assessed by this more comprehensive approach.

Different Stability-Indicating Chromatographic Techniques for the Determination of Netobimin

PubMed Central

Ramadan, Nesrin K.; Mohamed, Afaf O.; Shawky, Sara E.; Salem, Maissa Y.

2012-01-01

Two simple, accurate, and sensitive methods were developed for the determination of netobimin in the presence of its degradation product. Method (A) was an HPLC method, performed on C18 column using acetonitrile/methanol/0.01 M potassium dihydrogen phosphate (56 : 14 : 30 by volume) as a mobile phase with a flow rate of 0.5 mL/min. Detection was performed at 254 nm. Method (B) was a TLC method, using silica gel 60 F254 plates; the optimized mobile phase was toluene/methanol/chloroform/ammonium hydroxide (5 : 4 : 6 : 0.1 by volume). The spots were scanned densitometrically at 346 nm. Linearity ranges were 1–10 μg/mL for method (A) and 0.5–5 μg/band for method (B), and the mean percentage recoveries were 99.3 ± 0.7% and 99.7 ± 0.7% for methods (A) and (B), respectively. The proposed methods were found to be specific for netobimin in the presence of up to 90% of its degradation product. Statistical comparison between the results obtained by these methods and the manufacturer method was done, and no significance difference was obtained. PMID:22567566

Effects of zinc and DHA on the epigenetic regulation of human neuronal cells.

PubMed

Sadli, Nadia; Ackland, M Leigh; De Mel, Damitha; Sinclair, Andrew J; Suphioglu, Cenk

2012-01-01

Dietary intake of zinc and omega-3 fatty acids (DHA) have health benefits for a number of human diseases. However, the molecular basis of these health benefits remains unclear. Recently, we reported that zinc and DHA affect expression levels of histones H3 and H4 in human neuronal M17 cells. Here, using immunoblotting and densitometric analysis, we aimed to investigate the effect of zinc and DHA on post-translational modifications of histone H3 in M17 cells. In response to increase in zinc concentration, we observed increase in deacetylation, methylation and phosphorylation of H3 and decrease in acetylation. We also investigated the role of zinc in apoptosis, and found that zinc reduced the levels of the anti-apoptotic marker Bcl-2 while increasing the apoptotic marker caspase-3 levels, correlating with cell viability assays. Conversely, DHA treatment resulted in increase in acetylation of H3 and Bcl-2 levels and decrease in deacetylation, methylation, phosphorylation of H3 and caspase-3 levels, suggesting that DHA promotes gene expression and neuroprotection. Our novel findings show the opposing effects of zinc and DHA on the epigenetic regulation of human neuronal cells and highlight the potential benefit of dietary intake of DHA for management of neurodegenerative diseases. Copyright © 2012 S. Karger AG, Basel.

Comparison of an imaging heel quantitative ultrasound device (DTU-one) with densitometric and ultrasonic measurements.

PubMed

Diessel, E; Fuerst, T; Njeh, C F; Hans, D; Cheng, S; Genant, H K

2000-01-01

The purpose of this study was to evaluate a new imaging ultrasound scanner for the heel, the DTU-one (Osteometer MediTech, Denmark), by comparing quantitative ultrasound (QUS) results with bone mineral density (BMD) of the heel and femur from dual X-ray absorptiometry (DXA), and by comparing the DTU-one with another QUS device, the UBA 575+. The regions of interest in the DXA heel scan were matched with the regions evaluated by the two QUS devices. 134 healthy and 16 osteoporotic women aged 30-84 years old were enrolled in the study. In vivo short-term precision of the DTU-one for broadband ultrasound attenuation (BUA) and speed of sound (SOS) was 2.9% and 0.1%, respectively, and long-term precision was 3.8% and 0.2%, respectively. Highest correlations (r) between QUS and BMD measurements were achieved when comparing DTU-one results with BMD in matched regions of the DXA heel scan. Correlation coefficients (r) were 0.81 for BUA and SOS. Highest correlations with the UBA 575+ were 0.68 and 0.72, respectively. The comparison of BMD in different femoral sites with BUA and SOS (DTU-one) varied from 0.62 to 0.69 when including the entire study population. The correlation between BMD values within different sites of the femur tended to be higher (from r = 0.81 to 0.93). When comparing BUA with BUA and SOS with SOS on the two QUS devices, the absolute QUS values differed significantly. However, correlations were relatively high, with 0.76 for BUA and 0.82 for SOS. In conclusion, the results of the new quantitative ultrasound device, the DTU-one, are highly correlated (r = 0.8) with results obtained using the UBA 575+ and with BMD in the heel. The precision of the DTU-one is comparable to other QUS devices for BUA and is high for SOS.

Prevalence of osteoporosis, vertebral fractures and hypovitaminosis D in postmenopausal women living in a rural environment.

PubMed

Gómez-de-Tejada Romero, María-Jesús; Navarro Rodríguez, María-del-Carmen; Saavedra Santana, Pedro; Quesada Gómez, José-Manuel; Jódar Gimeno, Esteban; Sosa Henríquez, Manuel

2014-03-01

First, to study the difference between two groups of postmenopausal women living in different population centres (rural vs urban) in the prevalence of osteoporosis, fragility fractures and factors which may influence them: hypovitaminosis D, bone mineral density, coexistence of other diseases which predispose to their appearance; secondly, to observe the influence of low socioeconomic status, categorised as poverty. 1229 postmenopausal women were studied, of whom 390 (31.7%), were living in rural areas and 839 (68.3%), in urban areas. Data regarding risk factors related to osteoporosis were obtained, and, among other biochemical measures, 25 hydroxyvitamin D and parathyroid hormone were determined. Bone densitometry was carried out in the lumbar spine and proximal femur, as well as lateral X-rays of the dorsal and lumbar spine. The women who lived in rural areas were older, shorter, heavier and had a higher body mass index than those from urban areas. Among the women from rural areas there was a higher prevalence of poverty, and higher levels of obesity, arterial hypertension and diabetes mellitus were observed, as well as a higher prevalence of densitometric osteoporosis. The rural women had lower values of bone mineral density in the lumbar spine and a higher prevalence of vertebral fractures and hypovitaminosis D. The variables which were associated independently with living in rural areas were poverty, obesity, vertebral fractures, BMD in the lumbar spine and levels of 25 hydroxyvitamin D. In our study, postmenopausal women who live in rural populations have more poverty, lower values of vitamin D, lower BMD in the lumbar spine and a higher prevalence of vertebral fractures and of osteoporosis. The higher prevalence of obesity, arterial hypertension and diabetes mellitus observed in these women may be adjuvant factors, all fostered by their socioeconomic state of poverty. Copyright © 2014. Published by Elsevier Ireland Ltd.

Homocysteine, folate, and vitamin B12 levels and vertebral fracture risk in postmenopausal women.

PubMed

El Maghraoui, Abdellah; Ghozlani, Imad; Mounach, Aziza; Rezqi, Asmaa; Oumghar, Khalid; Achemlal, Lahsen; Bezza, Ahmed; Ouzzif, Zhor

2012-01-01

The objective of this study was to examine the influence of homocysteine, vitamin B(12), and folate on the prevalence of asymptomatic osteoporotic vertebral fractures (VFs) using vertebral fracture assessment (VFA) in postmenopausal women. The study cohort consisted of 188 consecutive postmenopausal women (mean age, weight, and body mass index of 57.9 ± 8.5 [41-91]yr, 74.4 ± 13.5 [38-150]kg, and 30.4 ± 5.2 [17.1-50.7]kg/m(2), respectively). Lateral VFA images and scans of the lumbar spine and proximal femur were obtained using a Lunar Prodigy Vision densitometer (GE Healthcare Inc., Waukesha, WI). VFs were defined using a combination of Genant's semiquantitative approach and morphometry. Fifty-eight (30.9%) patients had densitometric osteoporosis. VFs were identified using VFA in 76 (40.4%) patients: 61 women had grade 1 VFs and 15 had grade 2 or 3 VFs. No statistical difference was shown between the 3 groups (absence of VFs, VFs grade 1, and VFs grade 2/3) concerning the biological parameters. Comparison of patients according to quartiles of homocysteine levels showed that women in the highest quartile were older and had a lower bone mineral density (BMD); however, the prevalence of VFs was not statistically different from that of women in the other quartile groups. Stepwise regression analysis showed that homocysteine was not independently associated with the presence of VFs, which was mainly related to the osteoporotic status. Although a weak association was observed between hyperhomocysteinemia and low BMD and a trend to higher prevalence of grade 2/3 VFs was observed, our study did not confirm that homocysteine, vitamin B(12), and folate status are important determinants of prevalent asymptomatic VFs in postmenopausal women. Copyright © 2012 The International Society for Clinical Densitometry. Published by Elsevier Inc. All rights reserved.

Behavioral and anatomical characterization of the bilateral sciatic nerve chronic constriction (bCCI) injury: correlation of anatomic changes and responses to cold stimuli

PubMed Central

2010-01-01

Background Unilateral constrictive sciatic nerve injury (uCCI) is a common neuropathic pain model. However, the bilateral constrictive injury (bCCI) model is less well studied, and shows unique characteristics. In the present study, we sought to correlate effects of bCCI on nocifensive responses to cold and mechanical stimuli with selected dorsal horn anatomic markers. bCCI or sham ligation of both rat sciatic nerves were followed up to 90 days of behavioural testing. Additional rats sacrificed at 15, 30 and 90 days were used for anatomic analyses. Behavioural tests included hindpaw withdrawal responses to topical acetone, cold plate testing, an operant thermal preference task and hindpaw withdrawal thresholds to mechanical probing. Results All nocifensive responses to cold increased and remained enhanced for >45 days. Mechanical withdrawal thresholds decreased for 25 days only. Densitometric analyses of immunoperoxidase staining in the superficial dorsal horn at L4-5 revealed decreased cholecystokinin (CCK) staining at all times after bCCI, decreased mu opiate receptor (MOR) staining, maximal at 15 days, increased neuropeptide Y (NPY) staining only at days 15 and 30, and increased neurokinin-1 receptor (NK-1R) staining at all time points, maximal at 15 days. Correlation analyses at 45 days post-bCCI, were significant for individual rat nocifensive responses in each cold test and CCK and NK-1R, but not for MOR or NPY. Conclusions These results confirm the usefulness of cold testing in bCCI rats, a new approach using CCI to model neuropathic pain, and suggest a potential value of studying the roles of dorsal horn CCK and substance P in chronic neuropathic pain. Compared to human subjects with neuropathic pain, responses to cold stimuli in rats with bCCI may be a useful model of neuropathic pain. PMID:20105332

Behavioral and anatomical characterization of the bilateral sciatic nerve chronic constriction (bCCI) injury: correlation of anatomic changes and responses to cold stimuli.

PubMed

Datta, Sukdeb; Chatterjee, Koel; Kline, Robert H; Wiley, Ronald G

2010-01-27

Unilateral constrictive sciatic nerve injury (uCCI) is a common neuropathic pain model. However, the bilateral constrictive injury (bCCI) model is less well studied, and shows unique characteristics. In the present study, we sought to correlate effects of bCCI on nocifensive responses to cold and mechanical stimuli with selected dorsal horn anatomic markers. bCCI or sham ligation of both rat sciatic nerves were followed up to 90 days of behavioural testing. Additional rats sacrificed at 15, 30 and 90 days were used for anatomic analyses. Behavioural tests included hindpaw withdrawal responses to topical acetone, cold plate testing, an operant thermal preference task and hindpaw withdrawal thresholds to mechanical probing. All nocifensive responses to cold increased and remained enhanced for >45 days. Mechanical withdrawal thresholds decreased for 25 days only. Densitometric analyses of immunoperoxidase staining in the superficial dorsal horn at L4-5 revealed decreased cholecystokinin (CCK) staining at all times after bCCI, decreased mu opiate receptor (MOR) staining, maximal at 15 days, increased neuropeptide Y (NPY) staining only at days 15 and 30, and increased neurokinin-1 receptor (NK-1R) staining at all time points, maximal at 15 days. Correlation analyses at 45 days post-bCCI, were significant for individual rat nocifensive responses in each cold test and CCK and NK-1R, but not for MOR or NPY. These results confirm the usefulness of cold testing in bCCI rats, a new approach using CCI to model neuropathic pain, and suggest a potential value of studying the roles of dorsal horn CCK and substance P in chronic neuropathic pain. Compared to human subjects with neuropathic pain, responses to cold stimuli in rats with bCCI may be a useful model of neuropathic pain.

Expression and characterization of an N-truncated form of the NifA protein of Azospirillum brasilense.

PubMed

Nishikawa, C Y; Araújo, L M; Kadowaki, M A S; Monteiro, R A; Steffens, M B R; Pedrosa, F O; Souza, E M; Chubatsu, L S

2012-02-01

Azospirillum brasilense is a nitrogen-fixing bacterium associated with important agricultural crops such as rice, wheat and maize. The expression of genes responsible for nitrogen fixation (nif genes) in this bacterium is dependent on the transcriptional activator NifA. This protein contains three structural domains: the N-terminal domain is responsible for the negative control by fixed nitrogen; the central domain interacts with the RNA polymerase σ(54) co-factor and the C-terminal domain is involved in DNA binding. The central and C-terminal domains are linked by the interdomain linker (IDL). A conserved four-cysteine motif encompassing the end of the central domain and the IDL is probably involved in the oxygen-sensitivity of NifA. In the present study, we have expressed, purified and characterized an N-truncated form of A. brasilense NifA. The protein expression was carried out in Escherichia coli and the N-truncated NifA protein was purified by chromatography using an affinity metal-chelating resin followed by a heparin-bound resin. Protein homogeneity was determined by densitometric analysis. The N-truncated protein activated in vivo nifH::lacZ transcription regardless of fixed nitrogen concentration (absence or presence of 20 mM NH(4)Cl) but only under low oxygen levels. On the other hand, the aerobically purified N-truncated NifA protein bound to the nifB promoter, as demonstrated by an electrophoretic mobility shift assay, implying that DNA-binding activity is not strictly controlled by oxygen levels. Our data show that, while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription.

Expression and characterization of an N-truncated form of the NifA protein of Azospirillum brasilense

PubMed Central

Nishikawa, C.Y.; Araújo, L.M.; Kadowaki, M.A.S.; Monteiro, R.A.; Steffens, M.B.R.; Pedrosa, F.O.; Souza, E.M.; Chubatsu, L.S.

2012-01-01

Azospirillum brasilense is a nitrogen-fixing bacterium associated with important agricultural crops such as rice, wheat and maize. The expression of genes responsible for nitrogen fixation (nif genes) in this bacterium is dependent on the transcriptional activator NifA. This protein contains three structural domains: the N-terminal domain is responsible for the negative control by fixed nitrogen; the central domain interacts with the RNA polymerase σ54 factor and the C-terminal domain is involved in DNA binding. The central and C-terminal domains are linked by the interdomain linker (IDL). A conserved four-cysteine motif encompassing the end of the central domain and the IDL is probably involved in the oxygen-sensitivity of NifA. In the present study, we have expressed, purified and characterized an N-truncated form of A. brasilense NifA. The protein expression was carried out in Escherichia coli and the N-truncated NifA protein was purified by chromatography using an affinity metal-chelating resin followed by a heparin-bound resin. Protein homogeneity was determined by densitometric analysis. The N-truncated protein activated in vivo nifH::lacZ transcription regardless of fixed nitrogen concentration (absence or presence of 20 mM NH4Cl) but only under low oxygen levels. On the other hand, the aerobically purified N-truncated NifA protein bound to the nifB promoter, as demonstrated by an electrophoretic mobility shift assay, implying that DNA-binding activity is not strictly controlled by oxygen levels. Our data show that, while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription. PMID:22267004

Liver/kidney microsomal antibody type 1 and liver cytosol antibody type 1 concentrations in type 2 autoimmune hepatitis.

PubMed

Muratori, L; Cataleta, M; Muratori, P; Lenzi, M; Bianchi, F B

1998-05-01

Liver/kidney microsomal antibody type 1 (LKM1) and liver cytosol antibody type 1 (LC1) are the serological markers of type 2 autoimmune hepatitis (AIH). Since LKM1 and LC1 react against two distinct liver specific autoantigens (cytochrome P450IID6 (CYP2D6) and a 58 kDa cytosolic polypeptide respectively), the aim was to see whether LKM1 and LC1 concentrations correlate with liver disease activity. Twenty one patients with type 2 AIH were studied. All sera were tested by indirect immunofluorescence, counterimmunoelectrophoresis, and immunoblotting visualised by enhanced chemiluminescence. To evaluate LKM1 and LC1 levels, the 50 kDa microsomal reactivity (corresponding to CYP2D6) and the 58 kDa cytosolic reactivity were quantified by densitometric analysis. Seven patients were positive for LKM1, nine for LC1, and five for both. Serial serum samples at onset and during immunosuppressive treatment were analysed in 13 patients (four positive for LKM1, six positive for LC1 and three positive for both). During remission, LKM1 concentration remained essentially unchanged in six of seven patients, and decreased in only one. Conversely, in two of nine patients, LC1 was completely lost, and, in the remaining seven, LC1 concentration was reduced by more than 50%. After immunosuppression tapering or withdrawal, flare ups of liver necrosis ensued with increasing LC1 concentration, but not LKM1. LC1 concentration, at variance with that of LKM1, parallels liver disease activity, and its participation in the pathogenic mechanisms of liver injury can be hypothesised.

Liver/kidney microsomal antibody type 1 and liver cytosol antibody type 1 concentrations in type 2 autoimmune hepatitis

PubMed Central

Muratori, L; Cataleta, M; Muratori, P; Lenzi, M; Bianchi, F

1998-01-01

Background—Liver/kidney microsomal antibody type 1 (LKM1) and liver cytosol antibody type 1 (LC1) are the serological markers of type 2 autoimmune hepatitis (AIH). 
Aims—Since LKM1 and LC1 react against two distinct liver specific autoantigens (cytochrome P450IID6 (CYP2D6) and a 58 kDa cytosolic polypeptide respectively), the aim was to see whether LKM1 and LC1 concentrations correlate with liver disease activity. 
Patients—Twenty one patients with type 2 AIH were studied. 
Methods—All sera were tested by indirect immunofluorescence, counterimmunoelectrophoresis, and immunoblotting visualised by enhanced chemiluminescence. To evaluate LKM1 and LC1 levels, the 50 kDa microsomal reactivity (corresponding to CYP2D6) and the 58 kDa cytosolic reactivity were quantified by densitometric analysis. 
Results—Seven patients were positive for LKM1, nine for LC1, and five for both. Serial serum samples at onset and during immunosuppressive treatment were analysed in 13 patients (four positive for LKM1, six positive for LC1 and three positive for both). During remission, LKM1 concentration remained essentially unchanged in six of seven patients, and decreased in only one. Conversely, in two of nine patients, LC1 was completely lost, and, in the remaining seven, LC1 concentration was reduced by more than 50%. After immunosuppression tapering or withdrawal, flare ups of liver necrosis ensued with increasing LC1 concentration, but not LKM1. 
Conclusions—LC1 concentration, at variance with that of LKM1, parallels liver disease activity, and its participation in the pathogenic mechanisms of liver injury can be hypothesised. 

 Keywords: autoantibodies; immunoblotting; LKM1; LC1; immunosuppression PMID:9659171

Development and validation of HPTLC fingerprints of three species of Alpinia with biomarker Galangin.

PubMed

Upadhye, Anuradha S; Rajopadhye, Anagha; Dias, Lourelle

2018-01-16

Alpinia galanga (L.) Willd. commonly called as Rasna, Greater galangal or Kulinjan is a medicinally important rhizome used in Indian traditional system of medicine to cure a number of ailments. A. galanga is the main source of a galangin -a medicinally important flavanol which has a number of pharmacological properties viz. anti-mutagenic, and anti-inflammatory. Due to the high demand for the rhizome of A. galanga traders are now substituting it with rhizomes of A. calcarata and A. officinarum. The present study aims to develop high performance thin layer chromatographic (HPTLC) fingerprinting of A. galanga with its adulterants or substitutes and to quantify bioactive galangin present thereof. Methanolic extracts were obtained from rhizomes of the three species of Alpinia used for HPTLC analysis using silica gel 60 F254 plates and hexane: ethyl acetate: acetic acid (6.2: 2.8: 1.0 v/v/v); the densitometric analysis was performed at 272 nm. By comparison of Rf values and of the spectra of the bands with those of the standard galangin was identified in all three samples. HPTLC quantitative analysis of the methanolic extracts showed the decline trend in the quantity of the galangin in the three species of Alpinia as A. galanga (7.67 ± 0.36 mg/g) > A. officinarum (5.77 ± 0.71 mg/g) > A. calcarata (4.31 ± 0.44 mg/g). The HPTLC method was validated using International Conference on Harmonization (ICH) guidelines. The HPTLC method showed good linearity, recovery and high precision of biomarker. Rapid and reproducible method is useful for routine analysis of galangin and quality control of Alpinia galangal along with its adulterants or substitutes.

Quantification of steviol glycosides in food products, Stevia leaves and formulations by planar chromatography, including proof of absence for steviol and isosteviol.

PubMed

Wald, Julian P; Morlock, Gertrud E

2017-07-14

Steviol glycosides may degrade in food products under certain processing and storage conditions. Hence, a method was developed that separated in the same chromatographic run seven important steviol glycosides, and additionally as a sum parameter, their reported breakdown products steviol and isosteviol. Through derivatizations with the 2-naphthol and the primuline reagent, the detection was selective and inexpensive. In case needed, the baseline separation of steviol and isosteviol was also demonstrated after a plate cut and subsequent short development (two-step method). The HPTLC method was robust with regard to varying sample matrix loads, as the stationary phase was used only once. A high sample throughput was achieved, i.e. 23 separations were performed in parallel on one plate. The total analysis time took 1h (30min application, 15min separation and 15min derivatization/densitometry) leading to a calculated analysis time of 2.6min per sample. The solvent consumption was 8mL in total (0.4mL per analysis). HPTLC-ESI-MS was employed for confirmation of the results obtained. Mass spectra were recorded only from the zones of interest, and not from matrix or background, leading to decisive advantages, such as less need for MS cleaning. The optimized HPTLC method was shown to effectively support quality control, as marketed samples may be falsified with cheaper synthetic sweeteners, which was also demonstrated in this study. The accuracy of the densitometric quantification in HPTLC was considered as high, as standards and samples were separated on fresh adsorbent and detected simultaneously under identical conditions, which minimized the influence of errors. Finally, the Aliivibrio fischeri bioassay was employed to obtain information on bioactive compounds in Stevia leaf extracts. Copyright © 2017 Elsevier B.V. All rights reserved.

In vivo short-term precision of hip structure analysis variables in comparison with bone mineral density using paired dual-energy X-ray absorptiometry scans from multi-center clinical trials.

PubMed

Khoo, Benjamin C C; Beck, Thomas J; Qiao, Qi-Hong; Parakh, Pallav; Semanick, Lisa; Prince, Richard L; Singer, Kevin P; Price, Roger I

2005-07-01

Hip structural analysis (HSA) is a technique for extracting strength-related structural dimensions of bone cross-sections from two-dimensional hip scan images acquired by dual energy X-ray absorptiometry (DXA) scanners. Heretofore the precision of the method has not been thoroughly tested in the clinical setting. Using paired scans from two large clinical trials involving a range of different DXA machines, this study reports the first precision analysis of HSA variables, in comparison with that of conventional bone mineral density (BMD) on the same scans. A key HSA variable, section modulus (Z), biomechanically indicative of bone strength during bending, had a short-term precision percentage coefficient of variation (CV%) in the femoral neck of 3.4-10.1%, depending on the manufacturer or model of the DXA equipment. Cross-sectional area (CSA), a determinant of bone strength during axial loading and closely aligned with conventional DXA bone mineral content, had a range of CV% from 2.8% to 7.9%. Poorer precision was associated with inadequate inclusion of the femoral shaft or femoral head in the DXA-scanned hip region. Precision of HSA-derived BMD varied between 2.4% and 6.4%. Precision of DXA manufacturer-derived BMD varied between 1.9% and 3.4%, arising from the larger analysis region of interest (ROI). The precision of HSA variables was not generally dependent on magnitude, subject height, weight, or conventional femoral neck densitometric variables. The generally poorer precision of key HSA variables in comparison with conventional DXA-derived BMD highlights the critical roles played by correct limb repositioning and choice of an adequate and appropriately positioned ROI.

EMBRYONIC DEVELOPMENT AND A QUANTITATIVE MODEL OF PROGRAMMED DNA ELIMINATION IN MESOCYCLOPS EDAX (S. A. FORBES, 1891) (COPEPODA: CYCLOPOIDA)

PubMed Central

Clower, Michelle K.; Holub, Ashton S.; Smith, Rebecca T.; Wyngaard, Grace A.

2016-01-01

The highly programmed fragmentation of chromosomes and elimination of large amounts of nuclear DNA from the presomatic cell lineages (i.e., chromatin diminution), occurs in the embryos of the freshwater zooplankton Mesocyclops edax (S. A. Forbes, 1891) (Crustacea: Copepoda). The somatic genome is reorganized and reduced to a size five times smaller even though the germline genome remains intact. We present the first comprehensive, quantitative model of DNA content throughout embryogenesis in a copepod that possesses embryonic DNA elimination. We used densitometric image analysis to measure the DNA content of polar bodies, germline and somatic nuclei, and excised DNA “droplets.” We report: 1) variable DNA contents of polar bodies, some of which do not contain the amount corresponding to the haploid germline genome size; 2) presence of pronuclei in newly laid embryo sacs; 3) gonomeric chromosomes in the second to fourth cleavage divisions and in the primordial germ cell and primordial endoderm cell during the fifth cleavage division; 4) timing of early embryonic cell stages, elimination of DNA, and divisions of the primordial germ cell and primordial endoderm cell at 22°C; and 5) persistence of a portion of the excised DNA “droplets” throughout embryogenesis. DNA elimination is a trait that spans multiple embryonic stages and a knowledge of the timing and variability of the associated cytological events with DNA elimination will promote the study of the molecular mechanisms involved in this trait. We propose the “genome yolk hypothesis” as a functional explanation for the persistence of the eliminated DNA that might serve as a resource during postdiminution cleavage divisions. PMID:27857452

CT Metrics of Airway Disease and Emphysema in Severe COPD

PubMed Central

Kim, Woo Jin; Silverman, Edwin K.; Hoffman, Eric; Criner, Gerard J.; Mosenifar, Zab; Sciurba, Frank C.; Make, Barry J.; Carey, Vincent; Estépar, Raúl San José; Diaz, Alejandro; Reilly, John J.; Martinez, Fernando J.; Washko, George R.

2009-01-01

Background: CT scan measures of emphysema and airway disease have been correlated with lung function in cohorts of subjects with a range of COPD severity. The contribution of CT scan-assessed airway disease to objective measures of lung function and respiratory symptoms such as dyspnea in severe emphysema is less clear. Methods: Using data from 338 subjects in the National Emphysema Treatment Trial (NETT) Genetics Ancillary Study, densitometric measures of emphysema using a threshold of −950 Hounsfield units (%LAA-950) and airway wall phenotypes of the wall thickness (WT) and the square root of wall area (SRWA) of a 10-mm luminal perimeter airway were calculated for each subject. Linear regression analysis was performed for outcome variables FEV1 and percent predicted value of FEV1 with CT scan measures of emphysema and airway disease. Results: In univariate analysis, there were significant negative correlations between %LAA-950 and both the WT (r = −0.28, p = 0.0001) and SRWA (r = −0.19, p = 0.0008). Airway wall thickness was weakly but significantly correlated with postbronchodilator FEV1% predicted (R = −0.12, p = 0.02). Multivariate analysis showed significant associations between either WT or SRWA (β = −5.2, p = 0.009; β = −2.6, p = 0.008, respectively) and %LAA-950 (β = −10.6, p = 0.03) with the postbronchodilator FEV1% predicted. Male subjects exhibited significantly thicker airway wall phenotypes (p = 0.007 for WT and p = 0.0006 for SRWA). Conclusions: Airway disease and emphysema detected by CT scanning are inversely related in patients with severe COPD. Airway wall phenotypes were influenced by gender and associated with lung function in subjects with severe emphysema. PMID:19411295

Measurement of body fat and hydration of the fat-free body in health and disease

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

Streat, S.J.; Beddoe, A.H.; Hill, G.L.

1985-06-01

Body fat mass, fat-free body mass and body water are basic components of body composition which are used in nutritional and metabolic studies and in patient care. A method of measuring total body fat (TBF), fat-free mass (FFM) and its hydration (TBW/FFM) involving prompt gamma in vivo neutron activation analysis (IVNAA) and tritium dilution has been compared with the more traditional methods of densitometry and skinfold anthropometry in 36 normal volunteers, and with skinfold anthropometry in 56 patients presenting for nutritional support. While the mean values of TBF were in reasonable agreement for the three methods in normals it wasmore » founds that skinfold anthropometry underestimated TBF relative to the IVNAA/tritium method by, on average, 3.0 kg (19%) in patients. Furthermore, the ranges of values in normals of the ratio TBW/FFM for the anthropometric (0.62 to 0.80) and densitometric (0.65 to 0.80) methods were much wider than the range for the IVNAA/tritium method (0.69 to 0.76), in which TBW was measured by tritium dilution in all cases. In the patients, the ranges of this ratio were 0.52 to 0.90 for the anthropometric method and 0.67 to 0.82 for the IVNAA/tritium method; clearly anthropometry yields values of TBW/FFM which are outside accepted biological limits. On the basis of these findings, ranges of TBW/FFM are suggested for both normal adults (0.69 to 0.75) and patients requiring nutritional support (0.67 to 0.83). Finally it is concluded that the IVNAA/tritium method is a suitable method for measuring TBF and FFM and particularly so when body composition is abnormal.« less

Stability-Indicating HPTLC Method for Studying Stress Degradation Behavior of Sulbutiamine HCl

PubMed Central

Farid, Nehal F.; Abdelwahab, Nada S.

2016-01-01

Sulbutiamine (SUL) is an ester of thiazides with neurotropic action. A new stability indicating HPTLC method has been developed and validated for the determination of SUL in the presence of different degradation products. The drug was subjected to different stress conditions following ICH strategy such as hydrolytic degradation (neutral, alkaline and acidic hydrolysis), oxidation, photodegradation and dry heat degradation. The drug demonstrated degradation under all decomposition conditions except neutral hydrolysis and dry heat, where the drug was completely degraded with 0.1 N NaOH, 1 N HCl and 30% H2O2 while it was partially degradaed by 0.1 N HCl, 3% H2O2 and UV light. Structure elucidation of the resulting degradation products was performed using ESI-Q-MS–MS. A well-defined peak for SUL was obtained at Rf = 0.46 and was completely separated from all obtained degradation products. Chromatographic separation was carried out on HPTLC aluminum plates precoated with silica gel 60 F254 using acetone–methylene chloride–ammonia buffer (pH 8.5 ± 0.2) (7:3:0.5, v/v) as a developing system. Densitometric scanning of the separated peaks was performed at 254 nm. System suitability testing parameters were calculated to ascertain the quality performance of the developed method. The method was validated with respect to USP guidelines regarding accuracy, precision, specificity, robustness and ruggedness. Good correlation coefficients were achieved in the range of 0.4–5.0 µg/band, and the limit of detection and limit of quantitation were found to be 0.11 and 0.33 µg/band, respectively. The utility of the suggested method was verified by application to Arcalion forte® tablets where no interference from additives was found. PMID:26759487

Identification of avocado (Persea americana) root proteins induced by infection with the oomycete Phytophthora cinnamomi using a proteomic approach.

PubMed

Acosta-Muñiz, Carlos H; Escobar-Tovar, Lina; Valdes-Rodríguez, Silvia; Fernández-Pavia, Silvia; Arias-Saucedo, Luis J; de la Cruz Espindola Barquera, Maria; Gómez Lim, Miguel Á

2012-01-01

Avocado root rot, caused by Phytophthora cinnamomi, is the most important disease that limits avocado production. A proteomic approach was employed to identify proteins that are upregulated by infection with P. cinnamomi. Different proteins were shown to be differentially expressed after challenge with the pathogen by two-dimensional (2-D) gel electrophoresis. A densitometric evaluation of protein expression indicated differential regulation during the time-course analyzed. Some proteins induced in response to the infection were identified by standard peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry and sequencing by MALDI LIFT-TOF/TOF tandem mass spectrometry. Of the 400 protein spots detected on 2-D gels, 21 seemed to change in abundance by 3 hours after infection. Sixteen proteins were upregulated, 5 of these were only detected in infected roots and 11 showed an increased abundance. Among the differentially expressed proteins identified are homologs to isoflavone reductase, glutathione S-transferase, several abscisic acid stress-ripening proteins, cinnamyl alcohol dehydrogenase, cinnamoyl-CoA reductase, cysteine synthase and quinone reductase. A 17.3-kDa small heat-shock protein and a glycine-rich RNA-binding protein were identified as downregulated. Our group is the first to report on gene induction in response to oomycete infection in roots from avocado, using proteomic techniques. Copyright © Physiologia Plantarum 2011.

Densitometric tomography using the measurement of muon flux

NASA Astrophysics Data System (ADS)

Hivert, F.; Busto, J.; Brunner, J.; Salin, P.; Gaffet, S.

2013-12-01

The knowledge of the subsurface properties is essentially obtained by geophysical methods, e.g. seismic imaging, electric prospection or gravimetry. The present work develops a recent method to investigate the in situ density of rocks using atmospheric the muon flux measurement , its attenuation depending on the rock density and thickness. This new geophysical technique have been mainly applied in volcanology (Lesparre N., 2011) using scintillator detectors. The present project (T2DM2) aims to realize underground muons flux measurements in order to characterizing the rock massif density variations above the LSBB underground research facility in Rustrel (France). The muon flux will be measure with a new Muon telescope instrumentation using Micromegas detectors in Time Projection Chambers (TPC) configuration. The first step of the work presented considers the muon flux simulation using the Gaisser model, for the interactions between muons and atmospheric particles, and the MUSIC code (Kudryavtsev V. A., 2008) for the muons/rock interactions. The results show that the muon flux attenuation caused by density variations are enough significant to be observed until around 500 m depth and for period of time in the order of one month. Such a duration scale and depth of investigation is compatible with the duration of the water transfer processes involved within the Karst unsaturated zone where LSBB is located. Our work now concentrates on the optimization of the spatial distribution of detectors that will be deployed in future.

Analysis of cytoskeletal proteins in posterior capsule opacification after implantation of acrylic and hydrogel intraocular lenses.

PubMed

Matsushima, Hiroyuki; Mukai, Kouichiro; Obara, Yoshitaka; Yoshida, Shinichiro; Clark, John I

2004-01-01

To analyze selected lens cytoskeletal proteins in posterior capsule opacification (PCO) 2 weeks after intraocular lens (IOL) implantation in rabbits. Department of Ophthalmology, Dokkyo University School of Medicine, Tochigi, Japan. Eight 10-week-old albino rabbits were prepared and anesthetized for phacoemulsification and aspiration of the crystalline lens and implantation of an acrylic or a hydrogel IOL. Two weeks postoperatively, the rabbits were killed and the IOLs removed for immunohistochemistry. Deparaffinized tissue sections were processed with antibodies against alpha-smooth muscle actin (alpha-SMA) and beta-crystallin to observe the types of PCO with the 2 IOL types. The proteins in the PCO tissue and the normal lens were homogenized, centrifuged, and analyzed using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) densitometric analysis and Western immunoblotting for actin and vimentin. Immunohistochemistry demonstrated a fibroblastic cell type expressing alpha-SMA and partial regeneration of epithelial cells, resulting in a lenticular structure that stained irregularly for beta-crystallin. The immunoreactivity of fibroblast-like cells to beta-crystallin appeared weaker than that of the regenerated lenticular structure. SDS-PAGE showed variability in the content of cytoskeletal proteins in the insoluble fractions of the PCO. Degradation of the cytoskeletal components was greater with the acrylic IOL than with the hydrogel IOL. Cytoskeletal proteins expressed during the formation of PCO and IOL implantation may have potential as therapeutic target proteins to improve the biocompatibility of IOLs.

HPTLC Method for the Determination of Paracetamol, Pseudoephedrine and Loratidine in Tablets and Human Plasma.

PubMed

Farid, Nehal Fayek; Abdelaleem, Eglal A

2016-04-01

A sensitive, accurate and selective high performance thin layer chromatography (HPTLC) method was developed and validated for the simultaneous determination of paracetamol (PAR), its toxic impurity 4-aminophenol (4-AP), pseudoephedrine HCl (PSH) and loratidine (LOR). The proposed chromatographic method has been developed using HPTLC aluminum plates precoated with silica gel 60 F254 using acetone-hexane-ammonia (4:5:0.1, by volume) as a developing system followed by densitometric measurement at 254 nm for PAR, 4-AP and LOR, while PSH was scanned at 208 nm. System suitability testing parameters were calculated to ascertain the quality performance of the developed chromatographic method. The method was validated with respect to USP guidelines regarding accuracy, precision and specificity. The method was successfully applied for the determination of PAR, PSH and LOR in ATSHI(®) tablets. The three drugs were also determined in plasma by applying the proposed method in the ranges of 0.5-6 µg/band, 1.6-12 µg/band and 0.4-2 µg/band for PAR, PSH and LOR, respectively. The results obtained by the proposed method were compared with those obtained by a reported HPLC method, and there was no significance difference between both methods regarding accuracy and precision. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

CT-assisted reverse engineering for aging aircraft resupply

NASA Astrophysics Data System (ADS)

Yancey, Robert N.

1998-03-01

The service life of military aircraft is being extended beyond original design intents. When the C-130 is eventually retired, it will have been in service for 79 years, well beyond its planned life expectancy of 40 years. Similarly, the KC-135s are presently expected to remain operational for 86 years, and the B-52 for 94 years. Not only are inventories of parts in short supply, but it is necessary to acquire parts no one expected to replace. The first step in any resupply activity is the creation of a data package. If nor computer-aided design (CAD) model exists, the demands of modern electronic commerce dictate than one be created. Creating a CAD model of an existing part is referred to as 'reverse engineering.' Computed tomography (CT) offers an ideal way to obtain metrology data critical to reveres engineering activities. Industrial CT systems have progressed to the point where they can nondestructively measure part dimensions at an accuracy competitive with coordinate measuring machines and a speed competitive with laser scanners. However, of the existing methods, only CT can nondestructively dimension interior surfaces, and only CT has the ability to densitometrically quantify the internal state of materials. The use of CT to help create CAD models for resupply efforts will be described and examples presented. Additionally, examples will be presented how the CT-created CAD models were then sued to fabricate replacement parts for aging systems.

Qualitative and quantitative analysis of hyaluronan oligosaccharides with high performance thin layer chromatography using reagent-free derivatization on amino-modified silica and electrospray ionization-quadrupole time-of-flight mass spectrometry coupling on normal phase.

PubMed

Rothenhöfer, Martin; Scherübl, Rosmarie; Bernhardt, Günther; Heilmann, Jörg; Buschauer, Armin

2012-07-27

Purified oligomers of hyalobiuronic acid are indispensable tools to elucidate the physiological and pathophysiological role of hyaluronan degradation by various hyaluronidase isoenzymes. Therefore, we established and validated a novel sensitive, convenient, rapid, and cost-effective high performance thin layer chromatography (HPTLC) method for the qualitative and quantitative analysis of small saturated hyaluronan oligosaccharides consisting of 2-4 hyalobiuronic acid moieties. The use of amino-modified silica as stationary phase allows a simple reagent-free in situ derivatization by heating, resulting in a very low limit of detection (7-19 pmol per band, depending on the analyzed saturated oligosaccharide). By this derivatization procedure for the first time densitometric quantification of the analytes could be performed by HPTLC. The validated method showed a quantification limit of 37-71 pmol per band and was proven to be superior in comparison to conventional detection of hyaluronan oligosaccharides. The analytes were identified by hyphenation of normal phase planar chromatography to mass spectrometry (TLC-MS) using electrospray ionization. As an alternative to sequential techniques such as high performance liquid chromatography (HPLC) and capillary electrophoresis (CE), the validated HPTLC quantification method can easily be automated and is applicable to the analysis of multiple samples in parallel. Copyright © 2012 Elsevier B.V. All rights reserved.

Quantification of birefringence readily measures the level of muscle damage in zebrafish

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

Berger, Joachim, E-mail: Joachim.Berger@Monash.edu; Sztal, Tamar; Currie, Peter D.

2012-07-13

Highlights: Black-Right-Pointing-Pointer Report of an unbiased quantification of the birefringence of muscle of fish larvae. Black-Right-Pointing-Pointer Quantification method readily identifies level of overall muscle damage. Black-Right-Pointing-Pointer Compare zebrafish muscle mutants for level of phenotype severity. Black-Right-Pointing-Pointer Proposed tool to survey treatments that aim to ameliorate muscular dystrophy. -- Abstract: Muscular dystrophies are a group of genetic disorders that progressively weaken and degenerate muscle. Many zebrafish models for human muscular dystrophies have been generated and analysed, including dystrophin-deficient zebrafish mutants dmd that model Duchenne Muscular Dystrophy. Under polarised light the zebrafish muscle can be detected as a bright area in anmore » otherwise dark background. This light effect, called birefringence, results from the diffraction of polarised light through the pseudo-crystalline array of the muscle sarcomeres. Muscle damage, as seen in zebrafish models for muscular dystrophies, can readily be detected by a reduction in the birefringence. Therefore, birefringence is a very sensitive indicator of overall muscle integrity within larval zebrafish. Unbiased documentation of the birefringence followed by densitometric measurement enables the quantification of the birefringence of zebrafish larvae. Thereby, the overall level of muscle integrity can be detected, allowing the identification and categorisation of zebrafish muscle mutants. In addition, we propose that the establish protocol can be used to analyse treatments aimed at ameliorating dystrophic zebrafish models.« less

Validated HPTLC Method for Dihydrokaempferol-4'-O-glucopyranoside Quantitative Determination in Alcea Species.

PubMed

Abdel Salam, Nabil A; Ghazy, Nabila M; Shawky, Eman; Sallam, Shimaa M; Shenouda, Mary L

2018-07-01

Dihydrokaempferol-4'-O-glucopyranoside, a flavanonol glucoside, is the major compound in the flower of Alcea rosea L. which possesses significant antioxidant and anticancer activity against HepG-2 cell line and thus can be considered a marker compound for A. rosea L. We attempted to establish a new simple, validated high-performance thin-layer chromatographic (HPTLC) method for the quantitation of dihydrokaempferol-4'-O-glucopyranoside to help in the standardization of the hydroalcoholic extracts of A. rosea L. flowers and to evaluate the best method for its extraction from the plant material. The separation was carried out on an HPTLC aluminum plate pre-coated with silica gel 60F-254, eluted with ethyl acetate-methanol-water-acetic acid (30:5:4:0.15 v/v). Densitometric scanning was performed using a Camag TLC scanner III, at 295 nm. A linear relationship was obtained between the concentrations (0.9-3.6 mg) and peak areas with the correlation coefficient (r) of 0.9971 ± 0.0002. The percentage relative standard deviations of intra-day and inter-day precisions were 0.22-1.45 and 0.49-1.66, respectively. The percentage w/w of dihydrokaempferol-4'-O-glucopyranoside in the flowers of A. rosea L. after maceration and sonication for 15 min was found to be 0.733 g/100 g and 0.928 g/100 g, respectively.

Body composition of university students by hydrostatic weighing and skinfold measurement.

PubMed

Jürimäe, T; Jagomägi, G; Lepp, T

1992-12-01

The body composition of 124 male and 70 female Tartu University students was measured by three different methods: hydrostatic weighing by maximal expiration, hydrostatic weighing by maximal inspiration and subcutaneous fat thickness measurements. Our results show that the proposed body density measuring method by maximal expiration is simple, reliable and applicable not only in indoor swimming pools but in field conditions as well. The second new hydrostatic weighing apparatus is more comfortable for the subjects where the body density is measured at maximal inspiration. The mean body density of males was somewhat higher when measured by the maximal inspiration (1.066 +/- 0.012 g.ml-1) than when measured by the maximal expiration (1.063 +/- 0.009 g.ml-1, p < 0.05). For females, on the contrary, the maximal expiration method (1.044 +/- 0.010 g.ml-1) yielded a higher body density value than when measured by the maximal inspiration (1.040 +/- 0.011 g.ml-1, p > 0.05). The body fat percentage measured by skinfold thickness correlated significantly with the body fat percentage calculated by body density by maximal expiration (males r = 0.420, females r = 0.531) and inspiration (males r = 0.507, females r = 0.663). We must conclude that the presented two methods of measuring body density offer new possibilities for densitometric analysis without the need for expensive laboratory equipment.

DXA and pQCT predict pertrochanteric and not femoral neck fracture load in a human side-impact fracture model.

PubMed

Gebauer, Matthias; Stark, Olaf; Vettorazzi, Eik; Grifka, Joachim; Püschel, Klaus; Amling, Michael; Beckmann, Johannes

2014-01-01

The validity of dual energy X-ray absorptiometry (DXA) and peripheral quantitative computed tomography (pQCT) measurements as predictors of pertrochanteric and femoral neck fracture loads was compared in an experimental simulation of a fall on the greater trochanter. 65 proximal femora were harvested from patients at autopsy. All specimens were scanned with use of DXA for areal bone mineral density and pQCT for volumetric densities at selected sites of the proximal femur. A three-point bending test simulating a side-impact was performed to determine fracture load and resulted in 16 femoral neck and 49 pertrochanteric fractures. Regression analysis revealed that DXA BMD trochanter was the best variable at predicting fracture load of pertrochanteric fractures with an adjusted R(2) of 0.824 (p < 0.0001). There was no correlation between densitometric parameters and the fracture load of femoral neck fractures. A significant correlation further was found between body weight, height, femoral head diameter, and neck length on the one side and fracture load on the other side, irrespective of the fracture type. Clinically, the DXA BMD trochanter should be favored and integrated routinely as well as biometric and geometric parameters, particularly in elderly people with known osteoporosis at risk for falls. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

CALIPSO: an interactive image analysis software package for desktop PACS workstations

NASA Astrophysics Data System (ADS)

Ratib, Osman M.; Huang, H. K.

1990-07-01

The purpose of this project is to develop a low cost workstation for quantitative analysis of multimodality images using a Macintosh II personal computer. In the current configuration the Macintosh operates as a stand alone workstation where images are imported either from a central PACS server through a standard Ethernet network or recorded through video digitizer board. The CALIPSO software developed contains a large variety ofbasic image display and manipulation tools. We focused our effort however on the design and implementation ofquantitative analysis methods that can be applied to images from different imaging modalities. Analysis modules currently implemented include geometric and densitometric volumes and ejection fraction calculation from radionuclide and cine-angiograms Fourier analysis ofcardiac wall motion vascular stenosis measurement color coded parametric display of regional flow distribution from dynamic coronary angiograms automatic analysis ofmyocardial distribution ofradiolabelled tracers from tomoscintigraphic images. Several of these analysis tools were selected because they use similar color coded andparametric display methods to communicate quantitative data extracted from the images. 1. Rationale and objectives of the project Developments of Picture Archiving and Communication Systems (PACS) in clinical environment allow physicians and radiologists to assess radiographic images directly through imaging workstations (''). This convenient access to the images is often limited by the number of workstations available due in part to their high cost. There is also an increasing need for quantitative analysis ofthe images. During thepast decade

Postnatal development of glycine receptor subunits α1, α2, α3, and β immunoreactivity in multiple brain stem respiratory-related nuclear groups of the rat

PubMed Central

LIU, QIULI; WONG-RILEY, MARGARET T.T.

2013-01-01

The respiratory system is immature at birth and significant development occurs postnatally. A critical period of respiratory development occurs in rats around postnatal days 12-13, when enhanced inhibition dominates over suppressed excitation. The mechanisms underlying the heightened inhibition are not fully understood. The present study tested our hypothesis that the inhibition is marked by a switch in glycine receptor subunits from neonatal to adult form around the critical period. An in-depth immunohistochemical and single neuron optical densitometric study was undertaken on four respiratory-related nuclear groups (the pre-Bötzinger complex, nucleus ambiguus, hypoglossal nucleus, and ventrolateral subnucleus of solitary tract nucleus), and a non-respiratory cuneate nucleus in P2-21 rats. Our data revealed that in the respiratory-related nuclear groups: (1) the expressions of GlyRα2 and GlyRα3 were relatively high at P2, but declined after 1-1½ weeks to their lowest levels at P21; (2) the expression of GlyRα1 increased with age and reached significance at P12; and (3) the expression of GlyRβ rose from P2 to P12 followed by a slight decline until P21. No distinct increase in GlyRα1 at P12 was noted in the cuneate nucleus. Thus, there is a switch in dominance of expression from neonatal GlyRα2/α3 to the adult GlyRα1 and a heightened expression of GlyRα1 around the critical period in all respiratory-related nuclear groups, thereby supporting enhanced inhibition at that time. The rise in the expression of GlyRβ around P12 indicates that it plays an important role in forming the mature heteropentameric glycine receptors in these brain stem nuclear groups. PMID:24080401

High physiological prolactin induced by pituitary transplantation decreases BMD and BMC in the femoral metaphysis, but not in the diaphysis of adult female rats.

PubMed

Thongchote, Kanogwun; Charoenphandhu, Narattaphol; Krishnamra, Nateetip

2008-02-01

High physiological prolactin (PRL) stimulated intestinal calcium absorption and renal calcium uptake in mammals. Previous histomorphometric study revealed a significant increase in bone turnover in the trabecular part of the PRL-exposed long (cortical) bone; however, whole-bone densitometric analysis was unable to demonstrate such effect. We therefore studied differential changes in bone mineral density (BMD) and contents (BMC) of the femoral diaphysis and metaphysis in adult female rats exposed to high PRL induced by anterior pituitary (AP) transplantation. The estrogen-dependent effects of PRL on the femur were also investigated. We found that chronic exposure to PRL had no effect on BMD or BMC of the femoral diaphysis, which represented the cortical part of the long bone. It is interesting that 7 weeks after an AP transplantation, BMD and BMC of the femoral metaphysis were significantly decreased by 8% and 14%, respectively. Ovariectomy (Ovx) for 2, 5, and 7 weeks also decreased BMD and BMC in the femoral metaphysis, but not in the diaphysis. However, the AP transplantation plus Ovx (AP+Ovx) produced no additive effects. Nevertheless, 2.5 microg/kg 17beta-estradiol (E2) supplementation abolished the osteopenic effects of both Ovx and AP+Ovx on the femur. As for the L5-6 vertebrae, BMD and BMC were not affected by PRL exposure, but were significantly decreased by Ovx and AP+Ovx, and such decreases were completely prevented by E2 supplementation. It could be concluded that high physiological PRL induced a significant osteopenia in the trabecular part, i.e., the metaphysis, of the femora of adult female rats in an estrogen-dependent manner. Since PRL had no detectable effect on the vertebrae, the effects of PRL on bone appeared to be site-specific.

Canine epidermal lipid sampling by skin scrub revealed variations between different body sites and normal and atopic dogs

PubMed Central

2014-01-01

Background Previously, we evaluated a minimally invasive epidermal lipid sampling method called skin scrub, which achieved reproducible and comparable results to skin scraping. The present study aimed at investigating regional variations in canine epidermal lipid composition using the skin scrub technique and its suitability for collecting skin lipids in dogs suffering from certain skin diseases. Eight different body sites (5 highly and 3 lowly predisposed for atopic lesions) were sampled by skin scrub in 8 control dogs with normal skin. Additionally, lesional and non-lesional skin was sampled from 12 atopic dogs and 4 dogs with other skin diseases by skin scrub. Lipid fractions were separated by high performance thin layer chromatography and analysed densitometrically. Results No significant differences in total lipid content were found among the body sites tested in the control dogs. However, the pinna, lip and caudal back contained significantly lower concentrations of ceramides, whereas the palmar metacarpus and the axillary region contained significantly higher amounts of ceramides and cholesterol than most other body sites. The amount of total lipids and ceramides including all ceramide classes were significantly lower in both lesional and non-lesional skin of atopic dogs compared to normal skin, with the reduction being more pronounced in lesional skin. The sampling by skin scrub was relatively painless and caused only slight erythema at the sampled areas but no oedema. Histological examinations of skin biopsies at 2 skin scrubbed areas revealed a potential lipid extraction from the transition zone between stratum corneum and granulosum. Conclusions The present study revealed regional variations in the epidermal lipid and ceramide composition in dogs without skin abnormalities but no connection between lipid composition and predilection sites for canine atopic dermatitis lesions. The skin scrub technique proved to be a practicable sampling method for canine epidermal lipids, revealed satisfying results regarding alterations of skin lipid composition in canine atopic dermatitis and might be suitable for epidermal lipid investigations of further canine skin diseases. Although the ceramide composition should be unaffected by the deeper lipid sampling of skin scrub compared to other sampling methods, further studies are required to determine methodological differences. PMID:25012966

Validation of a Thin-Layer Chromatography for the Determination of Hydrocortisone Acetate and Lidocaine in a Pharmaceutical Preparation

PubMed Central

Dołowy, Małgorzata; Kulpińska-Kucia, Katarzyna; Pyka, Alina

2014-01-01

A new specific, precise, accurate, and robust TLC-densitometry has been developed for the simultaneous determination of hydrocortisone acetate and lidocaine hydrochloride in combined pharmaceutical formulation. The chromatographic analysis was carried out using a mobile phase consisting of chloroform + acetone + ammonia (25%) in volume composition 8 : 2 : 0.1 and silica gel 60F254 plates. Densitometric detection was performed in UV at wavelengths 200 nm and 250 nm, respectively, for lidocaine hydrochloride and hydrocortisone acetate. The validation of the proposed method was performed in terms of specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, and robustness. The applied TLC procedure is linear in hydrocortisone acetate concentration range of 3.75 ÷ 12.50 μg·spot−1, and from 1.00 ÷ 2.50 μg·spot−1 for lidocaine hydrochloride. The developed method was found to be accurate (the value of the coefficient of variation CV [%] is less than 3%), precise (CV [%] is less than 2%), specific, and robust. LOQ of hydrocortisone acetate is 0.198 μg·spot−1 and LOD is 0.066 μg·spot−1. LOQ and LOD values for lidocaine hydrochloride are 0.270 and 0.090 μg·spot−1, respectively. The assay value of both bioactive substances is consistent with the limits recommended by Pharmacopoeia. PMID:24526880

Validation of a thin-layer chromatography for the determination of hydrocortisone acetate and lidocaine in a pharmaceutical preparation.

PubMed

Dołowy, Małgorzata; Kulpińska-Kucia, Katarzyna; Pyka, Alina

2014-01-01

A new specific, precise, accurate, and robust TLC-densitometry has been developed for the simultaneous determination of hydrocortisone acetate and lidocaine hydrochloride in combined pharmaceutical formulation. The chromatographic analysis was carried out using a mobile phase consisting of chloroform+acetone+ammonia (25%) in volume composition 8:2:0.1 and silica gel 60F254 plates. Densitometric detection was performed in UV at wavelengths 200 nm and 250 nm, respectively, for lidocaine hydrochloride and hydrocortisone acetate. The validation of the proposed method was performed in terms of specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, and robustness. The applied TLC procedure is linear in hydrocortisone acetate concentration range of 3.75÷12.50  μg·spot(-1), and from 1.00÷2.50  μg·spot(-1) for lidocaine hydrochloride. The developed method was found to be accurate (the value of the coefficient of variation CV [%] is less than 3%), precise (CV [%] is less than 2%), specific, and robust. LOQ of hydrocortisone acetate is 0.198  μg·spot(-1) and LOD is 0.066  μg·spot(-1). LOQ and LOD values for lidocaine hydrochloride are 0.270 and 0.090  μg·spot(-1), respectively. The assay value of both bioactive substances is consistent with the limits recommended by Pharmacopoeia.

Effect of cisplatin on bone transport osteogenesis in dogs.

PubMed

Ehrhart, Nicole; Eurell, Jo Ann C; Tommasini, Matteo; Constable, Peter D; Johnson, Ann L; Feretti, Antonio

2002-05-01

To document effects of cisplatin on regenerate bone formation during the distraction and consolidation phases of bone transport osteogenesis. 10 skeletally mature hounds. Bone transport osteogenesis was performed to reconstruct a 3-cm defect in the radius of each dog. Five dogs were randomly selected to receive cisplatin (70 mg/m2, IV, q 21 d for 4 cycles), and 5 were administered saline (0.9% NaCl) solution. Bone mineral density was measured by use of dual-energy x-ray absorptiometry (DEXA) on days 24, 55, and 90 after surgery. Dogs were euthanatized 90 days after surgery. Histomorphometry was performed on nondecalcified sections of regenerate bone. Bone mineral density and histomorphometric indices of newly formed bone were compared between groups. Densitometric differences in regenerate bone mineral density were not detected between groups at any time period. Cisplatin-treated dogs had decreased mineralized bone volume, decreased percentage of woven bone volume, decreased percentage of osteoblast-covered bone, increased porosity, and increased percentage of osteoblast-covered surfaces, compared with values for control dogs. Lamellar bone volume and osteoid volume did not differ significantly between groups. Regenerate bone will form and remodel during administration of cisplatin. Results of histomorphometric analysis suggest that bone formation and resorption may be uncoupled in cisplatin-treated regenerate bone as a result of increased osteoclast activity or delayed secondary bone formation during remodeling. These histomorphometric differences were modest in magnitude and did not result in clinically observable complications or decreased bone mineral density as measured by use of DEXA.

Hypoxia-ischemia brain damage disrupts brain cholesterol homeostasis in neonatal rats.

PubMed

Yu, Z; Li, S; Lv, S H; Piao, H; Zhang, Y H; Zhang, Y M; Ma, H; Zhang, J; Sun, C K; Li, A P

2009-08-01

The first 3 weeks of life is the peak time of oligodendrocytes development and also the critical period of cholesterol increasing dramatically in central nervous system in rats. Neonatal hypoxia-ischemia (HI) brain damage happening in this period may disturb the brain cholesterol balance as well as white matter development. To test this hypothesis, postnatal day 7 (P7) Sprague-Dawley rats were subjected to HI insult. Cholesterol concentrations from brain and plasma were measured. White matter integrity was evaluated by densitometric analysis of myelin basic protein (MBP) immunostaining and electron microscopy. Brain TNF-alpha and IL-6 levels were also measured. HI-induced brain cholesterol, but not the plasma cholesterol, levels decreased significantly during the first three days after HI compared with naïve and sham operated rats (p<0.05). Obvious hypomyelination was indicated by marked reductions in MBP immunostaining on both P10 and P14 (p<0.01) and less and thinner myelinated axons were detected on P21 by electron microscopy observation. High expressions of brain TNF-alpha and IL-6 12 h after HI (p<0.05) were also observed. The present work provides evidence that HI insult destroyed brain cholesterol homeostasis, which might be important in the molecular pathology of hypoxic-ischemic white matter injury. Proinflammatory cytokines insulting oligodendrocytes, may cause cholesterol unbalance. Furthermore, specific therapeutic interventions to maintain brain cholesterol balance may be effective for the recovery of white matter function. Georg Thieme Verlag KG Stuttgart New York.

High-contrast x-ray microtomography in dental research

NASA Astrophysics Data System (ADS)

Davis, Graham; Mills, David

2017-09-01

X-ray microtomography (XMT) is a well-established technique in dental research. The technique has been used extensively to explore the complex morphology of the root canal system, and to qualitatively and quantitatively evaluate root canal instrumentation and filling efficacy in extracted teeth; enabling different techniques to be compared. Densitometric information can be used to identify and map demineralized tissue resulting from tooth decay (caries) and, in extracted teeth, the method can be used to evaluate different methods of excavation. More recently, high contrast XMT is being used to investigate the relationship between external insults to teeth and the pulpal reaction. When such insults occur, fluid may flow through dentinal tubules as a result of cracking or porosity in enamel. Over time, there is an increase in mineralization along the paths of the tubules from the pulp to the damaged region in enamel and this can be visualized using high contrast XMT. The scanner used for this employs time-delay integration to minimize the effects of detector inhomogeneity in order to greatly increase the upper limit on signal-to-noise ratio that can be achieved with long exposure times. When enamel cracks are present in extracted teeth, the presence of these pathways indicates that the cracking occurred prior to extraction. At high contrast, growth lines are occasionally seen in deciduous teeth which may have resulted from periods of maternal illness. Various other anomalies in mineralization resulting from trauma or genetic abnormalities can also be investigated using this technique.

Stability indicating high performance thin-layer chromatographic method for simultaneous estimation of pantoprazole sodium and itopride hydrochloride in combined dosage form

PubMed Central

Bageshwar, Deepak; Khanvilkar, Vineeta; Kadam, Vilasrao

2011-01-01

A specific, precise and stability indicating high-performance thin-layer chromatographic method for simultaneous estimation of pantoprazole sodium and itopride hydrochloride in pharmaceutical formulations was developed and validated. The method employed TLC aluminium plates precoated with silica gel 60F254 as the stationary phase. The solvent system consisted of methanol:water:ammonium acetate; 4.0:1.0:0.5 (v/v/v). This system was found to give compact and dense spots for both itopride hydrochloride (Rf value of 0.55±0.02) and pantoprazole sodium (Rf value of 0.85±0.04). Densitometric analysis of both drugs was carried out in the reflectance–absorbance mode at 289 nm. The linear regression analysis data for the calibration plots showed a good linear relationship with R2=0.9988±0.0012 in the concentration range of 100–400 ng for pantoprazole sodium. Also, the linear regression analysis data for the calibration plots showed a good linear relationship with R2=0.9990±0.0008 in the concentration range of 200–1200 ng for itopride hydrochloride. The method was validated for specificity, precision, robustness and recovery. Statistical analysis proves that the method is repeatable and selective for the estimation of both the said drugs. As the method could effectively separate the drug from its degradation products, it can be employed as a stability indicating method. PMID:29403710

Stability indicating high performance thin-layer chromatographic method for simultaneous estimation of pantoprazole sodium and itopride hydrochloride in combined dosage form.

PubMed

Bageshwar, Deepak; Khanvilkar, Vineeta; Kadam, Vilasrao

2011-11-01

A specific, precise and stability indicating high-performance thin-layer chromatographic method for simultaneous estimation of pantoprazole sodium and itopride hydrochloride in pharmaceutical formulations was developed and validated. The method employed TLC aluminium plates precoated with silica gel 60F 254 as the stationary phase. The solvent system consisted of methanol:water:ammonium acetate; 4.0:1.0:0.5 (v/v/v). This system was found to give compact and dense spots for both itopride hydrochloride ( R f value of 0.55±0.02) and pantoprazole sodium ( R f value of 0.85±0.04). Densitometric analysis of both drugs was carried out in the reflectance-absorbance mode at 289 nm. The linear regression analysis data for the calibration plots showed a good linear relationship with R 2 =0.9988±0.0012 in the concentration range of 100-400 ng for pantoprazole sodium. Also, the linear regression analysis data for the calibration plots showed a good linear relationship with R 2 =0.9990±0.0008 in the concentration range of 200-1200 ng for itopride hydrochloride. The method was validated for specificity, precision, robustness and recovery. Statistical analysis proves that the method is repeatable and selective for the estimation of both the said drugs. As the method could effectively separate the drug from its degradation products, it can be employed as a stability indicating method.

The BPAQ: a bone-specific physical activity assessment instrument.

PubMed

Weeks, B K; Beck, B R

2008-11-01

A newly developed bone-specific physical activity questionnaire (BPAQ) was compared with other common measures of physical activity for its ability to predict parameters of bone strength in healthy, young adults. The BPAQ predicted indices of bone strength at clinically relevant sites in both men and women, while other measures did not. Only certain types of physical activity (PA) are notably osteogenic. Most methods to quantify levels of PA fail to account for bone relevant loading. Our aim was to examine the ability of several methods of PA assessment and a new bone-specific measure to predict parameters of bone strength in healthy adults. We recruited 40 men and women (mean age 24.5). Subjects completed the modifiable activity questionnaire, Bouchard 3-day activity record, a recently published bone loading history questionnaire (BLHQ), and wore a pedometer for 14 days. We also administered our bone-specific physical activity questionnaire (BPAQ). Calcaneal broadband ultrasound attenuation (BUA) (QUS-2, Quidel) and densitometric measures (XR-36, Norland) were examined. Multiple regression and correlation analyses were performed on the data. The current activity component of BPAQ was a significant predictor of variance in femoral neck bone mineral density (BMD), lumbar spine BMD, and whole body BMD (R(2) = 0.36-0.68, p < 0.01) for men, while the past activity component of BPAQ predicted calcaneal BUA (R(2) = 0.48, p = 0.001) for women. The BPAQ predicted indices of bone strength at skeletal sites at risk of osteoporotic fracture while other PA measurement tools did not.

Effect of immersion on lung capacities and volumes: implications for the densitometric estimation of relative body fat.

PubMed

Withers, R T; Hamdorf, P A

1989-01-01

Immersion of 18 male subjects in water caused a 20.4% (787 ml) increase (P less than 0.05) in the mean inspiratory capacity (IC) whereas there were no changes (P greater than 0.05) in tidal volume (VT) and the frequency of respiration. All the means for the other pulmonary variables decreased (P less than 0.05) by varying amounts: total lung capacity (TLC) = 8.4% (599 ml), vital capacity (VC) = 5.5% (308 ml), functional residual capacity (FRC) = 42.6% (1386 ml), expiratory reserve volume (ERV) = 61.9% (1095 ml) and residual volume (RV) = 19.7% (292 ml). Variation of only the RV in the body density (BD) formula from which the percentage body fat (%BF) is estimated resulted in a significantly (P less than 0.05) lower mean of 15.2% BF for the RV in air (means = 1482 ml) compared with that of 17.1% BF for the RV in water (means = 1190 ml). All but one of the subjects exhibited a smaller RV in water than in air; the six largest differences were equivalent to 2.4-5.1% BF. These results indicate that the net effect of the hydrostatic pressure (decreases RV), pulmonary vascular engorgement (decreases RV) and diminished compliance (increases RV) is to reduce the ventilated RV. It is therefore advisable to measure the RV when the subject is immersed in order to minimize error in the determination of BD and hence the estimation of % BF.

Urinary exosomes in the diagnosis of Gitelman and Bartter syndromes.

PubMed

Corbetta, Samuele; Raimondo, Francesca; Tedeschi, Silvana; Syrèn, Marie-Louise; Rebora, Paola; Savoia, Andrea; Baldi, Lorenza; Bettinelli, Alberto; Pitto, Marina

2015-04-01

Gitelman syndrome (GS) and Bartter syndrome (BS) are hereditary salt-losing tubulopathies (SLTs) resulting from defects of renal proteins involved in electrolyte reabsorption, as for sodium-chloride cotransporter (NCC) and furosemide-sensitive sodium-potassium-chloride cotransporter (NKCC2) cotransporters, affected in GS and BS Type 1 patients, respectively. Currently, definitive diagnosis is obtained through expensive and time-consuming genetic testing. Urinary exosomes (UE), nanovesicles released by every epithelial cell facing the urinary space, represent an ideal source of markers for renal dysfunction and injury, because UE molecular composition stands for the cell of origin. On these assumptions, the aim of this work is to evaluate the relevance of UE for the diagnosis of SLTs. UE were purified from second morning urines collected from 32 patients with genetically proven SLTs (GS, BS1, BS2 and BS3 patients), 4 with unclassified SLTs and 22 control subjects (age and sex matched). The levels of NCC and NKCC2 were evaluated in UE by SDS-PAGE/western blotting with specific antibodies. Due to their location on the luminal side of tubular cells, NCC and NKCC2 are well represented in UE proteome. The NCC signal is significantly decreased/absent in UE of Gitelman patients compared with control subjects (Mann-Whitney t-test, P < 0.001) and, similarly, the NKCC2 in those of Bartter type 1 (P < 0.001). The difference in the levels of the two proteins allows recognition of Gitelman and Bartter type 1 patients from controls and, combined with clinical data, from other Bartter patients. Moreover, the receiver operating characteristic curve analysis using UE NCC densitometric values showed a good discriminating power of the test comparing GS patients versus controls and BS patients (area under the curve value = 0.92; sensitivity 84.2% and specificity 88.6%). UE phenotyping may be useful in the diagnosis of GS and BS, thus providing an alternative/complementary, urine-based diagnostic tool for SLT patient recognition and a diagnostic guidance in complex cases. © The Author 2014. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved.

Clodronate: new directions of use.

PubMed

Frediani, Bruno; Bertoldi, Ilaria

2015-01-01

Clodronate is the father of bisphosphonates. For over three decades it has been subject of study in biological and clinical areas, proving to be an extremely interesting molecule from different points of view. It has been the first drug for osteoporosis that can be administered pulsatorily (once every 15 days or once a week). This, along with good tolerability, has been the first cause of its success, when there were no solid data in literature about its antifracture efficacy. There are three published studies that prove its antifracture effect: two by McCloskey published in 2004 and 2007 on BMR, and our study about fracture prevention in corticosteroids OP. In these studies a dose of 800 mg/day orally administered or 100 mg/week I.M. was used, and they are basically the same if you consider that clodronate absorption, orally administered, is on average 1.9%. However, a series of works where higher doses were used (1600 mg orally administered) with greater effectiveness on bone mass, especially in higher risk populations, lead us to consider the use of 200 mg i.m. formulation. First of all, we proved densitometric equivalence of 200 mg i.m./14 days and 100 mg i.m./week in a first study; then, in a second study, we proved a greater densitometric efficacy of 200 mg/week compared to 100 mg/week, clearer at femoral level, where the drug had not proven to prevent femoral fracture because of inadequate bone mass increase at that level. Moreover, as for ibandronate case, monthly dose was doubled compared to pivotal trial, in order to maximize the effects on femoral bone mass and therefore prevent femoral fractures. Consequently, on the basis of the risk envelope, whether it is identified according to BMD and the presence of one risk factor at least or more correctly identified through risk chart (FRAX or DeFRA), you can put forward a differential use of 100 mg i.m. and 200 mg i.m., weekly, "off-label" or every 14 days, adjusting doses in relation to fracture risk and painful symptoms gravity, as well as improving its ease of use and therefore assist compliance. Common experience and clinical and biological works have proved that clodronate has an analgesic effect that can be increased by doubling the doses. The analgesic effect is present not only with patients with fractures, but also with patients suffering from osteoarthritis or arthritis. Therefore, the drug would fit well in the therapeutic program of rheumatic patient, also because of its symptomatic effects. Clodronate at small doses (2 mg) could also have protective effects on cartilage (introduction of intra-articular formulation is expected) and at 10-100 fold higher doses it has certainly anti-inflammatory effects and more specifically antimacrophage and anticytokine effects (IL-1, IL-6, TNFalpha, PGE). These effects are amplified by putting clodronate in monolayer liposomes. This drug, therefore, has to be considered as adjuvant in arthritis therapy, whose origin can be linked to a strong osteoclastic activation caused by an increase of cytokines and the RANKL/OPG relationship. It is clear that clodronate can work on cytokine at first and on osteoclastic effector in the end. The drug has been used "off-label" for decades intravenously in complex regional pain syndrome (CRPS type 1) in relation to schemes that change according to different Authors and according to cumulative doses ranging from 3 to 5 g. The introduction on the market of the 200 mg i.m. formulation could allow to get more practical but equally effective schemes. For example, we used this scheme: 200 mg/day for 10 days and then 200 mg every other day for 20 days (cumulative dose of 4 g in a month). Said scheme can be repeated in the following months in particular cases. Results, as for efficacy and lack of relapses, show that clodronate is the leader drug for this syndrome. In recent years, relationship between costs and benefits has started to matter, especially after the creation of some algorithms, such as FRAX, that let us choose patients with a higher fracture risk in 10 years, and after pharmaceutic-economic models that let us calculate FRAX intervention threshold, on the basis of drug price and monitoring, antifracture efficacy, quality of life and how much a community can or wants to spend. In this respect, a sub-analysis of McCloskey's study on people over the age of 75, conducted on 3974 subjects, shows that clodronate is more efficient with patients with a higher fracture risk, calculated according to FRAX. Furthermore, another study by McCloskey revealed that, for a 100-pound/year drug (very similar to clodronate), 'cost effective' intervention threshold is about 7-10%. In conclusion, clodronate prevents fractures, decrease osteo-articular pain, is easy to handle, tolerable and had a great cost/benefit relationship.

Postnatal development of Na+-K+-2Cl− co-transporter 1 (NKCC1) and K+-Cl−co-transporter 2 (KCC2) immunoreactivity in multiple brain stem respiratory nuclei of the rat

PubMed Central

Liu, Qiuli; Wong-Riley, Margaret T.T.

2012-01-01

Previously, we reported that in rats, GABAA and glycine receptor immunoreactivity increased markedly in multiple brain stem respiratory nuclei around postnatal days (P) 12–13, a critical period when abrupt neurochemical, metabolic, ventilatory, and electrophysiological changes occur in the respiratory network and when the system is under greater inhibition than excitation. Since Na+-K+-2Cl− co-transporter 1 (NKCC1) and K+-Cl− co-transporter 2 (KCC2) play pivotal roles in determining the responses of GABAA and glycine receptors, we hypothesized that NKCC1 and KCC2 undergo significant changes during the critical period. An in-depth immunohistochemical and single neuron optical densitometric study of neurons in seven respiratory-related nuclei (the pre-Bötzinger complex [PBC], nucleus ambiguus [Amb], hypoglossal nucleus [XII], ventrolateral subnucleus of solitary tract nucleus [NTSVL], retrotrapezoid nucleus/parafacial respiratory group [RTN/pFRG], dorsal motor nucleus of the vagus nerve [DMNX], and inferior olivary nucleus [IO]) and a non-respiratory cuneate nucleus (CN, an internal control) was undertaken in P0–21 rats. Our data revealed that: (1) NKCC1 immunoreactivity exhibited a developmental decrease from P0 to P21 in all eight nuclei examined, being relatively high during the first 1½ postnatal weeks and decreased thereafter. The decrease was abrupt and statistically significant at P12 in the PBC, Amb, and XII; (2) KCC2 immunoreactivity in these eight nuclei showed a developmental increase from P0 to P21; and (3) the significant reduction in NKCC1 and the greater dominance of KCC2 around P12 in multiple respiratory nuclei of the brain stem may form the basis of an enhanced inhibition in the respiratory network during the critical period before the system stabilizes to a more mature state. PMID:22441038

Racial differences in the expression of inhibitors of apoptosis (IAP) proteins in extracellular vesicles (EV) from prostate cancer patients.

PubMed

Khan, Salma; Simpson, Jennifer; Lynch, James C; Turay, David; Mirshahidi, Saied; Gonda, Amber; Sanchez, Tino W; Casiano, Carlos A; Wall, Nathan R

2017-01-01

African-American men with prostate cancer typically develop more aggressive tumors than men from other racial/ethnic groups, resulting in a disproportionately high mortality from this malignancy. This study evaluated differences in the expression of inhibitors of apoptosis proteins (IAPs), a known family of oncoproteins, in blood-derived exosomal vesicles (EV) between African-American and European-American men with prostate cancer. The ExoQuick™ method was used to isolate EV from both plasma and sera of African-American (n = 41) and European-American (n = 31) men with prostate cancer, as well as from controls with no cancer diagnosis (n = 10). EV preparations were quantified by acetylcholinesterase activity assays, and assessed for their IAP content by Western blotting and densitometric analysis. Circulating levels of the IAP Survivin were evaluated by ELISA. We detected a significant increase in the levels of circulating Survivin in prostate cancer patients compared to controls (P<0.01), with the highest levels in African-American patients (P<0.01). African-American patients with prostate cancer also contained significantly higher amounts of EVs in their plasma (P<0.01) and sera (P<0.05) than European-American patients. In addition, EVs from African-American patients with prostate cancer contained significantly higher amounts of the IAPs Survivin (P<0.05), XIAP (P<0.001), and cIAP-2 (P<0.01) than EVs from European-American patients. There was no significant correlation between expression of IAPs and clinicopathological parameters in the two patient groups. Increased expression of IAPs in EVs from African-American patients with prostate cancer may influence tumor aggressiveness and contribute to the mortality disparity observed in this patient population. EVs could serve as reservoirs of novel biomarkers and therapeutic targets that may have clinical utility in reducing prostate cancer health disparities.

The G protein-coupled estrogen receptor 1 (GPER/GPR30) does not predict survival in patients with ovarian cancer

PubMed Central

2012-01-01

Background Even though ovarian tumors are not generally considered estrogen-sensitive, estrogens may still have an impact on ovarian tumor progression. The recently identified trans-membrane estrogen receptor GPER is involved in rapid estrogen signaling. Furthermore, it binds selective estrogen receptor modulators with agonistic effect, which could explain tamoxifen controversies. Methods GPER mRNA was assayed with quantitative real-time PCR (qPCR) in 42 primary ovarian tumors and 7 ovarian cancer cell lines. ERα and ERβ mRNA were analyzed for comparison. GPER protein was semi-quantified with densitometric scanning of Western blots and its tissue distribution analyzed with immunohistochemistry (IHC) in 40 ovarian tumors. In addition, IHC was evaluated in a tissue microarray (TMA) of 150 primary malignant ovarian tumors. Results All tumor samples contained GPER mRNA. The content of mRNA was not different between benign and malignant tumors, but one third of malignant samples over-expressed GPER mRNA. The content of ERα mRNA was higher in malignant than in benign tumors, whereas ERβ mRNA was higher in benign than in malignant tumors. GPER mRNA was detected in all seven ovarian cancer cell lines with highest levels in TOV21G and TOV112D cells. Similar expression pattern was seen for ERβ mRNA. Western blot demonstrated GPER protein in all tumor samples. Semi-quantification showed no difference between benign and malignant tumors, but about one third of malignant samples over-expressed GPER protein. GPER staining was localized mainly in epithelial cells. In the TMA study we found no correlation between GPER staining and clinical stage, histological grade or patient survival. Conclusions GPER mRNA as well as GPER protein is present in both benign and malignant ovarian tumor tissue. About one third of malignant tumors over-expressed both GPER mRNA and protein. This, however, correlated neither with histological or clinical parameters nor with patient survival. PMID:22424333

Pharmacologic antagonism of dopamine receptor D3 attenuates neurodegeneration and motor impairment in a mouse model of Parkinson's disease.

PubMed

Elgueta, Daniela; Aymerich, María S; Contreras, Francisco; Montoya, Andro; Celorrio, Marta; Rojo-Bustamante, Estefanía; Riquelme, Eduardo; González, Hugo; Vásquez, Mónica; Franco, Rafael; Pacheco, Rodrigo

2017-02-01

Neuroinflammation involves the activation of glial cells, which is associated to the progression of neurodegeneration in Parkinson's disease. Recently, we and other researchers demonstrated that dopamine receptor D3 (D3R)-deficient mice are completely refractory to neuroinflammation and consequent neurodegeneration associated to the acute intoxication with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). In this study we examined the therapeutic potential and underlying mechanism of a D3R-selective antagonist, PG01037, in mice intoxicated with a chronic regime of administration of MPTP and probenecid (MPTPp). Biodistribution analysis indicated that intraperitoneally administered PG01037 crosses the blood-brain barrier and reaches the highest concentration in the brain 40 min after the injection. Furthermore, the drug was preferentially distributed to the brain in comparison to the plasma. Treatment of MPTPp-intoxicated mice with PG01037 (30 mg/kg, administrated twice a week for five weeks) attenuated the loss of dopaminergic neurons in the substantia nigra pars compacta, as evaluated by stereological analysis, and the loss of striatal dopaminergic terminals, as determined by densitometric analyses of tyrosine hydroxylase and dopamine transporter immunoreactivities. Accordingly, the treatment resulted in significant improvement of motor performance of injured animals. Interestingly, the therapeutic dose of PG01037 exacerbated astrogliosis and resulted in increased ramification density of microglial cells in the striatum of MPTPp-intoxicated mice. Further analyses suggested that D3R expressed in astrocytes favours a beneficial astrogliosis with anti-inflammatory consequences on microglia. Our findings indicate that D3R-antagonism exerts a therapeutic effect in parkinsonian animals by reducing the loss of dopaminergic neurons in the nigrostriatal pathway, alleviating motor impairments and modifying the pro-inflammatory phenotype of glial cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effects of short-term weightlessness on inner ear carbonic anhydrase reactivity in fish - a parabolic aircraft flight study

NASA Astrophysics Data System (ADS)

Anken, Ralf; Hilbig, Reinhard; Weigele, Jochen; Anken, Ralf

We have shown earlier that some fish of a given batch reveal motion sickness (a kinetosis) at the transition from increased gravity (hypergravity, hg; centrifuge) to 1g earth gravity. Total macular carbonic anhydrase (CA)-reactivity as well as the difference in reactivities between left and right maculae (asymmetry) were significantly lower in normally swimming hg-animals as compared to the kinetotically behaving hg-fish. This result clearly indicated the existence of a regulatory mechanism, which adjusts otolithic calcium carbonate incorporation via CA- reactivity towards the gravity vector. Thus, we were prompted to investigate, whether fish swimming kinetotically under weightlessness also would reveal an asymmetric CA-reactivity. Therefore, larval cichlid fish (Oreochromis mossambicus) were subjected to parabolic aircraft flights. During the flights, the animals were videorecorded. Subsequently, fish were separated according to their respective swimming behaviour into normally and abnormally (kinetotic) moving individuals (the latter performed spinning movements, i.e., turns around their longitudinal axis). Finally, CA was localized histochemically and densitometrically determined in inner ear maculae. It was found that the total macular CA-reactivity did not differ between ground controls and kinetotically or normally swimming experimental fish. Asymmetry of CA- reactivity, however, was considerably higher in experimental animals as compared to the ground controls. No difference in asymmetry of CA-reactivity was obtained when comparing kinetotic with normally behaving individuals. These results indicate that parabolic flights do not affect CA-reactivity in general, possibly due to the relatively quickly alternating G-levels (g-profile of single parabola: 1g/1.8g/0.04g/1.8g/1g, performed in 70 seconds) in up to 30 parabolas per flight day. The high asymmetry of CA-reactivity in the experimental animals - irrespective of their behaviour - probably indicates an adaptation process, which has presumably been successful in the normally swimming samples. Acknowledgement: This work was financially supported by the German Aerospace Center (DLR) (FKZ: 50 WB 0527).

Protein size resolution in human eye lenses by dynamic light scattering after in vivo measurements: first analysis of a clinical validation

NASA Astrophysics Data System (ADS)

Dierks, Karsten; Dieckmann, Matthias; Niederstrasser, Dirk; Schwartz, R.; Wegener, Alfred R.

1995-01-01

Laser light source powers of our instrument LEPOSIAR are minimized so that dynamic light scattering (DLS) measurements can be conducted with lowest intensity levels on human eye lenses (3.2 mW/cm2) within measurement times of 3 to 5 seconds. We describe an extension of DLS and an eye lens characterization along the optical axis (OA), revealing the molecular size ranges together with distributions found in lens regions parallel to the OA. The microstructures of various lens regions are separated by the analyzed radius distributions reflecting the visco-elastic properties of the eye lens. Detailed analysis and applied statistical categorization of results are described. The data obtained by DLS allow for an objective interpretation of opacity occurrences on molecular size range which are related to refraction anomalies. Apart from changes in color and fluorescence properties, the refractive anomalies can be assumed as the origin of the cataract. The bimodal radius distributions are characterized as a function of patient ages varying from 9 to 85 years. Our clinical study on 42 subjects proves a size decrease of the monomeric fraction with age, whereby their relative frequency of occurrence decreases. The larger polymeric radius fractions which are detected in lenses of all subjects increase with their age. An increase of protein polymer size is likely to be linked to the decrease of the (gamma) -crystallin fraction in old eye lens nuclei. Our preliminary analysis of clinical results is corresponding to the chemical gradients parallel to the OA of the lenses which are reported by O. Hockwin after examination on extracted eye lenses. The normalization of the lens densitometric data derived after Scheimpflug- photography against the protein size fraction analysis by DLS is performed. As a comparison mean between both techniques on identical eye lenses, an absorption corrected densitometry is conducted for the first time.

Stability-Indicating HPTLC Method for Studying Stress Degradation Behavior of Sulbutiamine HCl.

PubMed

Farid, Nehal F; Abdelwahab, Nada S

2016-04-01

Sulbutiamine (SUL) is an ester of thiazides with neurotropic action. A new stability indicating HPTLC method has been developed and validated for the determination of SUL in the presence of different degradation products. The drug was subjected to different stress conditions following ICH strategy such as hydrolytic degradation (neutral, alkaline and acidic hydrolysis), oxidation, photodegradation and dry heat degradation. The drug demonstrated degradation under all decomposition conditions except neutral hydrolysis and dry heat, where the drug was completely degraded with 0.1 N NaOH, 1 N HCl and 30% H2O2 while it was partially degradaed by 0.1 N HCl, 3% H2O2 and UV light. Structure elucidation of the resulting degradation products was performed using ESI-Q-MS-MS. A well-defined peak for SUL was obtained at Rf = 0.46 and was completely separated from all obtained degradation products. Chromatographic separation was carried out on HPTLC aluminum plates precoated with silica gel 60 F254 using acetone-methylene chloride-ammonia buffer (pH 8.5 ± 0.2) (7:3:0.5, v/v) as a developing system. Densitometric scanning of the separated peaks was performed at 254 nm. System suitability testing parameters were calculated to ascertain the quality performance of the developed method. The method was validated with respect to USP guidelines regarding accuracy, precision, specificity, robustness and ruggedness. Good correlation coefficients were achieved in the range of 0.4-5.0 µg/band, and the limit of detection and limit of quantitation were found to be 0.11 and 0.33 µg/band, respectively. The utility of the suggested method was verified by application to Arcalion forte® tablets where no interference from additives was found. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Longitudinal effects of Parathyroid Hormone treatment on morphological, densitometric and mechanical properties of mouse tibia.

PubMed

Lu, Yongtao; Boudiffa, Maya; Dall'Ara, Enrico; Liu, Yue; Bellantuono, Ilaria; Viceconti, Marco

2017-11-01

The use of Parathyroid Hormone (PTH) as bone anabolic is limited due to cost-benefit assessments. Preclinical studies evaluating the effects of PTH on bone have reported variable and often contradictory results. Here, we have applied a new approach using a combination of in-vivo longitudinal µCT, image processing techniques and finite element models to monitor early local changes in the whole tibia (divided in 40 compartments) and mechanical properties of female C57BL/6J mice treated with PTH 1-34, compared to controls. Compared with standard 3D bone morphometric analysis, our new approach allowed detection of much smaller and localised changes in bone mineral content (BMC) at very early time points (1 week vs 3 weeks with standard methods) and showed that changes do not occur uniformly over time and across the anatomical space. Indeed, in the PTH treated mice, significant changes in BMC were observed in the medial and posterior sectors of the proximal tibia, a week after treatment, and in the medial sector of the tibia midshaft region a week later (p < 0.05). By the third week, two thirds of the regions showed significantly higher values of BMC (p < 0.05). The effect of PTH on bone regional volume is similar to that on BMC, but there is almost no effect of PTH on bone tissue mineral density. The differences in estimated mechanical properties became significant after three weeks of treatment (p < 0.05). These results provide the first evidence of an early and localised PTH effect on murine bone, and show that our novel partitioning approach, compared to the standard evaluation protocol, allows a more precise quantification of bone changes following treatment, which would facilitate preclinical testing of novel mono- and/or combination therapies throughout the bone. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Gamma amino butyric acid accumulation in medicinal plants without stress

PubMed Central

Anju, P.; Moothedath, Ismail; Rema Shree, Azhimala Bhaskaranpillai

2014-01-01

Introduction: Gamma amino butyric acid (GABA) is an important ubiquitous four carbon nonprotein amino acid with an amino group attached to gamma carbon instead of beta carbon. It exists in different organisms including bacteria, plants, and animals and plays a crucial role in humans by regulating neuronal excitability throughout the nervous system. It is directly responsible for the regulation of muscle tone and also effective in lowering stress, blood pressure, and hypertension. Aim and Objective: The aim of the study was to develop the fingerprint profile of selected medicinally and economically important plants having central nervous system (CNS) activity and to determine the quantity of GABA in the selected plants grown under natural conditions without any added stress. Materials and Methods: The high-performance thin layer chromatography analysis was performed on precoated silica gel plate 60F–254 plate (20 cm × 10 cm) in the form of bands with width 8 mm using Hamilton syringe (100 μl) using n-butanol, acetic acid, and water in the proportion 5:2:2 as mobile phase in a CAMAG chamber which was previously saturated for 30 min. CAMAG TLC scanner 3 was used for the densitometric scanning at 550 nm. Specific marker compounds were used for the quantification. Results and Conclusion: Among the screened medicinal plants, Zingiber officinale and Solanum torvum were found to have GABA. The percentage of GABA present in Z. officinale and S. torvum were found to be 0.0114% and 0.0119%, respectively. The present work confirmed that among the selected CNS active medicinal plants, only two plants contain GABA. We found a negative correlation with plant having CNS activity and accumulation of GABA. The GABA shunt is a conserved pathway in eukaryotes and prokaryotes but, although the role of GABA as a neurotransmitter in mammals is clearly established, its role in plants is still vague. PMID:25861139

Fluorophore-assisted carbohydrate electrophoresis for the determination of molecular mass of heparins and low-molecular-weight (LMW) heparins.

PubMed

Buzzega, Dania; Maccari, Francesca; Volpi, Nicola

2008-11-01

We report the use of fluorophore-assisted carbohydrate electrophoresis (FACE) to determine the molecular mass (M) values of heparins (Heps) and low-molecular-weight (LMW)-Hep derivatives. Hep are labeled with 8-aminonaphthalene-1,3,6-trisulfonic acid and FACE is able to resolve each fraction as a discrete band depending on their M. After densitometric acquisition, the migration distance of each Hep standard is acquired and the third-grade polynomial calibration standard curve is determined by plotting the logarithms of the M values as a function of migration ratio. Purified Hep samples having different properties, pharmaceutical Heps and various LMW-Heps were analyzed by both FACE and conventional high-performance size-exclusion liquid chromatography (HPSEC) methods. The molecular weight value on the top of the chromatographic peak (Mp), the number-average Mn, weight-average Mw and polydispersity (Mw/Mn) were examined by both techniques and found to be similar. This approach offers certain advantages over the HPSEC method. The derivatization process with 8-aminonaphthalene-1,3,6-trisulfonic acid is complete after 4 h so that many samples may be analyzed in a day also considering that multiple samples can be run simultaneously and in parallel and that a single FACE analysis requires approx. 15 min. Furthermore, FACE is a very sensitive method as it requires approx. 5-10 microg of Heps, about 10-100-fold lower than samples and standards used in HPSEC evaluation. Finally, the utilization of mini-gels allows the use of very low amounts of reagents with neither expensive equipment nor any complicated procedures having to be applied. This study demonstrates that FACE analysis is a sensitive method for the determination of the M values of Heps and LMW-Heps with possible utilization in virtually any kind of research and development such as quality control laboratories due to its rapid, parallel analysis of multiple samples by means of common and simple largely used analytical laboratory equipment.

Gut-bone axis response to dietary replacement of soybean meal with raw low-tannin faba bean seeds in broiler chickens.

PubMed

Tomaszewska, Ewa; Dobrowolski, Piotr; Klebaniuk, Renata; Kwiecień, Małgorzata; Tomczyk-Warunek, Agnieszka; Szymańczyk, Sylwia; Kowalik, Sylwester; Milczarek, Anna; Blicharski, Tomasz; Muszyński, Siemowit

2018-01-01

It seems that faba bean (FB) seeds could be a good protein-energy component in animal feed, but the presence of anti-nutritional substances limits their use as a substitute of soybean meal. The aim of the study was to examine the influence of different concentrations of raw, low-tannin, FB seeds on the gut-bone axis in Ross 308 broilers. One-day old chickens were randomly subjected to one of the 3 dietary treatments: the control group was fed standard diet based on soybean meal and without FB seeds, and two groups were fed 8%/15% and 16%/22% of raw low-tannin FB seeds in the starter and grower, respectively. On the 35th day, hematological and serum biochemical analyses as well histomorphometry of the small intestine and liver tissue and bone mechanical tests were performed. The diet type had no effect on the body weight gain and feed conversion ratio. However, the basal intestinal structures were significantly reduced in birds fed the lower concentration of FB. The enlargement of nerve plexuses was dependent on the concentration used in the diet and, additionally, on the kind of plexus and location in the intestinal tract. The liver was characterized by an increase in non-hepatocytes. There was no influence of the low-tannin FB seeds on most of the analyzed serum parameters in the 35-day-old broiler chickens, except the decreased concentration of total cholesterol and Ca in both experimental groups, triglycerides in group I, and P and uric acid in group II. Furthermore, the increasing concentration of the dietary low-tannin FB did not influence the activities of AspAT (except the group fed the higher amount of FB), ALAT, and LDH. The broiler chickens had no visible leg lesions and no problem in the locomotor function, but the tibiae were lighter mainly in birds fed the higher concentration of FB seeds. Geometric analysis revealed reduction of the cross section area and wall thickness, indicating a decline in the bone midshaft, which influenced the densitometric parameters and the results of mechanical tests. In conclusion, the dietary inclusion of raw low-tannin FB seeds had no negative effects on broiler growth, but disturbed the intestine structure and tibia characteristics. Therefore, all these negative effects necessitate additional examinations before inclusion of raw low-tannin FB seed into poultry diet.

Employment of High-Performance Thin-Layer Chromatography for the Quantification of Oleuropein in Olive Leaves and the Selection of a Suitable Solvent System for Its Isolation with Centrifugal Partition Chromatography.

PubMed

Boka, Vasiliki-Ioanna; Argyropoulou, Aikaterini; Gikas, Evangelos; Angelis, Apostolis; Aligiannis, Nektarios; Skaltsounis, Alexios-Leandros

2015-11-01

A high-performance thin-layer chromatographic methodology was developed and validated for the isolation and quantitative determination of oleuropein in two extracts of Olea europaea leaves. OLE_A was a crude acetone extract, while OLE_AA was its defatted residue. Initially, high-performance thin-layer chromatography was employed for the purification process of oleuropein with fast centrifugal partition chromatography, replacing high-performance liquid-chromatography, in the stage of the determination of the distribution coefficient and the retention volume. A densitometric method was developed for the determination of the distribution coefficients, KC = CS/CM. The total concentrations of the target compound in the stationary phase (CS) and in the mobile phase (CM) were calculated by the area measured in the high-performance thin-layer chromatogram. The estimated Kc was also used for the calculation of the retention volume, VR, with a chromatographic retention equation. The obtained data were successfully applied for the purification of oleuropein and the experimental results confirmed the theoretical predictions, indicating that high-performance thin-layer chromatography could be an important counterpart in the phytochemical study of natural products. The isolated oleuropein (purity > 95%) was subsequently used for the estimation of its content in each extract with a simple, sensitive and accurate high-performance thin-layer chromatography method. The best fit calibration curve from 1.0 µg/track to 6.0 µg/track of oleuropein was polynomial and the quantification was achieved by UV detection at λ 240 nm. The method was validated giving rise to an efficient and high-throughput procedure, with the relative standard deviation % of repeatability and intermediate precision not exceeding 4.9% and accuracy between 92% and 98% (recovery rates). Moreover, the method was validated for robustness, limit of quantitation, and limit of detection. The amount of oleuropein for OLE_A, OLE_AA, and an aqueous extract of olive leaves was estimated to be 35.5% ± 2.7, 51.5% ± 1.4, and 12.5% ± 0.12, respectively. Statistical analysis proved that the method is repeatable and selective, and can be effectively applied for the estimation of oleuropein in olive leaves' extracts, and could potentially replace high-performance liquid chromatography methodologies developed so far. Thus, the phytochemical investigation of oleuropein could be based on high-performance thin-layer chromatography coupled with separation processes, such as fast centrifugal partition chromatography, showing efficacy and credibility. Georg Thieme Verlag KG Stuttgart · New York.

Gut-bone axis response to dietary replacement of soybean meal with raw low-tannin faba bean seeds in broiler chickens

PubMed Central

Dobrowolski, Piotr; Klebaniuk, Renata; Kwiecień, Małgorzata; Tomczyk-Warunek, Agnieszka; Szymańczyk, Sylwia; Kowalik, Sylwester; Milczarek, Anna; Blicharski, Tomasz; Muszyński, Siemowit

2018-01-01

It seems that faba bean (FB) seeds could be a good protein-energy component in animal feed, but the presence of anti-nutritional substances limits their use as a substitute of soybean meal. The aim of the study was to examine the influence of different concentrations of raw, low-tannin, FB seeds on the gut-bone axis in Ross 308 broilers. One-day old chickens were randomly subjected to one of the 3 dietary treatments: the control group was fed standard diet based on soybean meal and without FB seeds, and two groups were fed 8%/15% and 16%/22% of raw low-tannin FB seeds in the starter and grower, respectively. On the 35th day, hematological and serum biochemical analyses as well histomorphometry of the small intestine and liver tissue and bone mechanical tests were performed. The diet type had no effect on the body weight gain and feed conversion ratio. However, the basal intestinal structures were significantly reduced in birds fed the lower concentration of FB. The enlargement of nerve plexuses was dependent on the concentration used in the diet and, additionally, on the kind of plexus and location in the intestinal tract. The liver was characterized by an increase in non-hepatocytes. There was no influence of the low-tannin FB seeds on most of the analyzed serum parameters in the 35-day-old broiler chickens, except the decreased concentration of total cholesterol and Ca in both experimental groups, triglycerides in group I, and P and uric acid in group II. Furthermore, the increasing concentration of the dietary low-tannin FB did not influence the activities of AspAT (except the group fed the higher amount of FB), ALAT, and LDH. The broiler chickens had no visible leg lesions and no problem in the locomotor function, but the tibiae were lighter mainly in birds fed the higher concentration of FB seeds. Geometric analysis revealed reduction of the cross section area and wall thickness, indicating a decline in the bone midshaft, which influenced the densitometric parameters and the results of mechanical tests. In conclusion, the dietary inclusion of raw low-tannin FB seeds had no negative effects on broiler growth, but disturbed the intestine structure and tibia characteristics. Therefore, all these negative effects necessitate additional examinations before inclusion of raw low-tannin FB seed into poultry diet. PMID:29590191

Section 9.1 new dosimeters. New dosimetry systems

NASA Astrophysics Data System (ADS)

McLaughlin, William L.

During the past two years there have been significant advances in several forms of radiation measurement systems for radiation processing, covering dose ranges of 1-10 6 Gy. Calorimeters as reference standards for both ionizing photon and electron fields have become well-established. In addition to the older ceric-cerous dosimetry solution analyzed potentiometrically, new liquid-phase dosimeters include those analyzed by spectrophotometry, e.g., improved forms of acidic aqueous solutions of K-Ag dichromate and organic radiochromic dye solutions. It has recently been demonstrated that by using certain refined sugars, e.g., D-(-) ribose, optical rotation response in aqueous solutions can be enhanced for dosimetry at doses > 10 4 Gy. There has been expanded development, use, and formulation (rods, tablets, and thin films) of the amino acid, alanine, as a solid-phase dosimeter analyzed by either ESR spectrometry or by glutamine or alanine spectrophotometry of complexes with ferric ion in the presence of a sulfonphthalein dye (xylenol orange). New commercial types of radiochromic plastic dosimeters, e.g., GafChromic TM, Riso B3 TM, GAMMACHROME YR TM, Radix TM, and Gammex TM, have been introduced and applied in practice. Improvements and broader use of optical waveguide dosimeters, e.g., Opti-Chromic TM, have also been reported, especially in food irradiation applications. Several novel dyed plastic dosimeters are available in large quantities and they lose color due to irradiation. An example is a dyed cellulosic thin film (ATC type DY-42 TM) which can be measured spectrophotometrically or densitometrically up to doses as high as 10 6 Gy.

Morphological alterations in the hippocampus of the Ts65Dn mouse model for Down Syndrome correlate with structural plasticity markers.

PubMed

Villarroya, Olga; Ballestín, Raúl; López-Hidalgo, Rosa; Mulet, Maria; Blasco-Ibáñez, José Miguel; Crespo, Carlos; Nacher, Juan; Gilabert-Juan, Javier; Varea, Emilio

2018-01-01

Down syndrome (DS) is the most common chromosomal aneuploidy. Although trisomy on chromosome 21 can display variable phenotypes, there is a common feature among all DS individuals: the presence of intellectual disability. This condition is partially attributed to abnormalities found in the hippocampus of individuals with DS and in the murine model for DS, Ts65Dn. To check if all hippocampal areas were equally affected in 4-5 month adult Ts65Dn mice, we analysed the morphology of dentate gyrus granule cells and cornu ammonis pyramidal neurons using Sholl method on Golgi-Cox impregnated neurons. Structural plasticity has been analysed using immunohistochemistry for plasticity molecules followed by densitometric analysis (Brain Derived Neurotrophic Factor (BDNF), Polysialylated form of the Neural Cell Adhesion Molecule (PSA-NCAM) and the Growth Associated Protein 43 (GAP43)). We observed an impairment in the dendritic arborisation of granule cells, but not in the pyramidal neurons in the Ts65Dn mice. When we analysed the expression of molecules related to structural plasticity in trisomic mouse hippocampus, we observed a reduction in the expression of BDNF and PSA-NCAM, and an increment in the expression of GAP43. These alterations were restricted to the regions related to dentate granule cells suggesting an interrelation. Therefore the impairment in dendritic arborisation and molecular plasticity is not a general feature of all Down syndrome principal neurons. Pharmacological manipulations of the levels of plasticity molecules could provide a way to restore granule cell morphology and function.

Densitometric thin-layer chromatographic determination of aescin in a herbal medicinal product containing Aesculus and Vitis dry extracts.

PubMed

Apers, Sandra; Naessens, Tania; Pieters, Luc; Vlietinck, Arnold

2006-04-21

A thin-layer chromatographic (TLC) method is developed to analyze the total saponin content, also referred to as the aescin content, in a herbal medicinal product (HMP) containing two dry extracts in capsules. The capsules contain 250 mg of Aesculus hippocastanum dry extract, 120 mg of Vitis vinifera dry extract and 50mg of excipients. After a purification step using C(18) solid phase extraction (SPE) cartridges, the samples are analyzed on a silica-gel HPTLC plate with the upper layer of a mixture of acetic acid/water/butanol (10/40/50 v/v/v) as the mobile phase. Spots are visualized by spraying with anisaldehyde reagent and heating the plate for 5-10 min (100-105 degrees C) and measured at a wavelength of 535 nm. This method, applicable for the quality control and stability investigation of both the Aesculus dry extract and HMP capsules thereof containing Vitis dry extract in combination with the Aesculus dry extract, is validated according to the International Conference on Harmonization (ICH) guidelines. The proposed assay method is specific for aescin in the presence of Vitis dry extract and formulation excipients. Analysis of stressed samples in forced degradation tests proves the method to be applicable for stability evaluation. The standard aescin curve is linear (r > 0.99) over a concentration range of 0.16-0.80 microg/spot. Recovery from the HMP capsules is statistically equal to 100%. The precision of the method with respect to time and concentration is acceptable, with relative standard deviation (RSD) values of 1.28 and 1.49%, respectively.

[Toward an anthropometric diagnosis of osteopenia and a biochemical diagnosis of osteoporoses].

PubMed

Cointry, Gustavo R; Capozza, Ricardo F; Ferretti, Jose L; Frost, Harold M

2003-01-01

The current (metabolic) conception of bone-weakening diseases regards bone strength as determined by a systemically-controlled "mineralized mass" which grows until it reaches a peak and then is lost at individually-specific rates. This concept disregards bone biomechanics. Skeletons are structures, it reaches of which depends on the stiffness and the spatial distribution rather than the volume of the calcified material. Rather than allowing a systemic regulation of their "mass" as a way to optimize their strength, bones autocontrol their stiffness by orienting bone formation and destruction as locally determined by the directional sensing, by osteocytes, of the strains caused by mechanical usage (gravity, muscle contractions). Bone mass and strength are just side products of that control. Endocrine-metabolic systems modulate non-directionally the work of bone cells as required for achieving a mineral equilibrium, despite the biomechanical controls, and can determine osteopenias and osteoporoses. Osteoporoses are not "intense osteopenias" (as per the current WHO's conception) but "osteopenic bone fragilities" (as recently stated by the NIH). The diagnosis of osteopenia is an anthropometric problem that can be solved densitometrically; but that of bone fragility is a biomechanical matter that requires evaluation of bone material's stiffness and distribution by other means ("resistometry"). For therapeutic purposes, osteopenias and osteoporoses should be also evaluated according to the relationship between bone mass or strength and muscle mass or strength in order to distinguish between "mechanical" (disuse) and "metabolic" etiologies (intrinsic bone lesion, or systemic disequilibrium), in which the bone/muscle proportionality tends to remain normal or to deteriorate, respectively.

Comparative distribution of sulphur, thiols and disulphides in the porcine stratum corneum.

PubMed

Meyer, W; Zschemisch, N H; Lehmann, H; Busche, R; Kunz, U

2005-01-01

Biochemical, histochemical and cytochemical analyses were used to determine the sulphur contents and the thiol and disulphide distribution in the stratum corneum (SC) of the wild boar (WB), a large domestic pig breed (DP) and the Goettingen miniature pig (GMP). The sulphur contents (% DW) were different in the three animal types (WB: 1.70-1.38 body, 0.54 ear; DP: 0.84-0.53 body, 0.50 ear; GMP: 2.28-2.51 body, 2.66 ear). The results of the histochemical analysis of SH- and -S-S- groups were clear, and densitometrical extinctions were highest in most body regions of the GMP for thiols and disulphides, followed by the DP for thiols, and the WB for disulphides. Absolute SC thickness was highest in the body of the GMP (62-80 mum), and generally lowest in the ear (20-38 mum) of all animal types. Relative SC thickness was the same for all animals in the body (40-66%), but lower in the ear (30%). Only -S-S- concentrations were correlated with SC thickness, and primarily in the GMP. Cytochemical analysis showed that high sulphur concentrations were obvious particularly in the CCE of corneal cells in the DP, as compared to the cytoplasm. Intracellular sulphur distribution was homogenous in the WB, and in the GMP, although in the latter at a higher concentration level. The results indicate breed-related effects on keratinisation in porcine corneal cells. Only the SC of the outer side of the ear of DP females is recommended as a model for humans.

High-performance Thin-layer Chromatographic-densitometric Quantification and Recovery of Bioactive Compounds for Identification of Elite Chemotypes of Gloriosa superba L. Collected from Sikkim Himalayas (India)

PubMed Central

Misra, Ankita; Shukla, Pushpendra Kumar; Kumar, Bhanu; Chand, Jai; Kushwaha, Poonam; Khalid, Md.; Singh Rawat, Ajay Kumar; Srivastava, Sharad

2017-01-01

Background: Gloriosa superba L. (Colchicaceae) is used as adjuvant therapy in gout for its potential antimitotic activity due to high colchicine(s) alkaloids. Objective: This study aimed to develop an easy, cheap, precise, and accurate high-performance thin-layer chromatographic (HPTLC) validated method for simultaneous quantification of bioactive alkaloids (colchicine and gloriosine) in G. superba L. and to identify its elite chemotype(s) from Sikkim Himalayas (India). Methods: The HPTLC chromatographic method was developed using mobile phase of chloroform: acetone: diethyl amine (5:4:1) at λmax of 350 nm. Results: Five germplasms were collected from targeted region, and on morpho-anatomical inspection, no significant variation was observed among them. Quantification data reveal that content of colchicine (Rf: 0.72) and gloriosine (Rf: 0.61) varies from 0.035%–0.150% to 0.006%–0.032% (dry wt. basis). Linearity of method was obtained in the concentration range of 100–400 ng/spot of marker(s), exhibiting regression coefficient of 0.9987 (colchicine) and 0.9983 (gloriosine) with optimum recovery of 97.79 ± 3.86 and 100.023% ± 0.01%, respectively. Limit of detection and limit of quantification were analyzed, respectively, as 6.245, 18.926 and 8.024, 24.316 (ng). Two germplasms, namely NBG-27 and NBG-26, were found to be elite chemotype of both the markers. Conclusion: The developed method is validated in terms of accuracy, recovery, and precision studies as per the ICH guidelines (2005) and can be adopted for the simultaneous quantification of colchicine and gloriosine in phytopharmaceuticals. In addition, this study is relevant to explore the chemotypic variability in metabolite content for commercial and medicinal purposes. SUMMARY An easy, cheap, precise, and accurate high performance thin layer chromatographic (HPTLC) validated method for simultaneous quantification of bioactive alkaloids (colchicine and gloriosine) in G. superba L.Five germplasms were collected from targeted region, and on morpho anatomical inspection, no significant variation was observed among themQuantification data reveal that content of colchicine (Rf: 0.72) and gloriosine (Rf: 0.61) varies from 0.035%–0.150% to 0.006%–0.032% (dry wt. basis)Two germplasms, namely NBG 27 and NBG 26, were found to be elite chemotype of both the markers. PMID:29142436

High-performance Thin-layer Chromatographic-densitometric Quantification and Recovery of Bioactive Compounds for Identification of Elite Chemotypes of Gloriosa superba L. Collected from Sikkim Himalayas (India).

PubMed

Misra, Ankita; Shukla, Pushpendra Kumar; Kumar, Bhanu; Chand, Jai; Kushwaha, Poonam; Khalid, Md; Singh Rawat, Ajay Kumar; Srivastava, Sharad

2017-10-01

Gloriosa superba L. (Colchicaceae) is used as adjuvant therapy in gout for its potential antimitotic activity due to high colchicine(s) alkaloids. This study aimed to develop an easy, cheap, precise, and accurate high-performance thin-layer chromatographic (HPTLC) validated method for simultaneous quantification of bioactive alkaloids (colchicine and gloriosine) in G. superba L. and to identify its elite chemotype(s) from Sikkim Himalayas (India). The HPTLC chromatographic method was developed using mobile phase of chloroform: acetone: diethyl amine (5:4:1) at λ max of 350 nm. Five germplasms were collected from targeted region, and on morpho-anatomical inspection, no significant variation was observed among them. Quantification data reveal that content of colchicine ( R f : 0.72) and gloriosine ( R f : 0.61) varies from 0.035%-0.150% to 0.006%-0.032% (dry wt. basis). Linearity of method was obtained in the concentration range of 100-400 ng/spot of marker(s), exhibiting regression coefficient of 0.9987 (colchicine) and 0.9983 (gloriosine) with optimum recovery of 97.79 ± 3.86 and 100.023% ± 0.01%, respectively. Limit of detection and limit of quantification were analyzed, respectively, as 6.245, 18.926 and 8.024, 24.316 (ng). Two germplasms, namely NBG-27 and NBG-26, were found to be elite chemotype of both the markers. The developed method is validated in terms of accuracy, recovery, and precision studies as per the ICH guidelines (2005) and can be adopted for the simultaneous quantification of colchicine and gloriosine in phytopharmaceuticals. In addition, this study is relevant to explore the chemotypic variability in metabolite content for commercial and medicinal purposes. An easy, cheap, precise, and accurate high performance thin layer chromatographic (HPTLC) validated method for simultaneous quantification of bioactive alkaloids (colchicine and gloriosine) in G. superba L.Five germplasms were collected from targeted region, and on morpho anatomical inspection, no significant variation was observed among themQuantification data reveal that content of colchicine (Rf: 0.72) and gloriosine (Rf: 0.61) varies from 0.035%-0.150% to 0.006%-0.032% (dry wt. basis)Two germplasms, namely NBG 27 and NBG 26, were found to be elite chemotype of both the markers.

Bisphosphonate effects in rat unloaded hindlimb bone loss model: three-dimensional microcomputed tomographic, histomorphometric, and densitometric analyses.

PubMed

Barou, O; Lafage-Proust, M H; Martel, C; Thomas, T; Tirode, F; Laroche, N; Barbier, A; Alexandre, C; Vico, L

1999-10-01

The effects of antiresorptive drugs on bone loss remain unclear. Using three-dimensional microtomography, dual X-ray/densitometry, and histomorphometry, we evaluated tiludronate effects in the bone loss model of immobilization in tail-suspended rats after 7, 13, and 23 days. Seventy-eight 12-week-old Wistar male rats were assigned to 13 groups: 1 baseline group, and for each time point, 1 control group treated with vehicle and three tail-suspended groups treated with either tiludronate (0.5 or 5 mg/kg) or vehicle, administered s. c. every other day, during the last week before sacrifice. In primary spongiosa (ISP), immobilization-induced bone loss plateaued after day 7 and was prevented by tiludronate. In secondary spongiosa (IISP), bone loss appeared at day 13 with a decrease in trabecular thickness and trabecular number (Tb.N) as assessed by three-dimensional microtomography. Osteoclastic parameters did not differ in tail-suspended rats versus control rats, whereas bone formation showed a biphasic pattern: after a marked decrease at day 7, osteoblastic activity and recruitment normalized at days 13 and 23, respectively. At day 23, the 80% decrease in bone mass was fully prevented by high-dose tiludronate with an increase in Tb.N without preventing trabecular thinning. In summary, at day 7, tiludronate prevented bone loss in ISP. After day 13, tiludronate prevented bone loss in ISP and IISP despite a further decrease in bone formation. Thus, the preventive effects of tiludronate in this model may be related to the alteration in bone modeling with an increase in Tb.N in ISP and subsequently in IISP.

Lobar Emphysema Distribution Is Associated With 5-Year Radiological Disease Progression.

PubMed

Boueiz, Adel; Chang, Yale; Cho, Michael H; Washko, George R; San José Estépar, Raul; Bowler, Russell P; Crapo, James D; DeMeo, Dawn L; Dy, Jennifer G; Silverman, Edwin K; Castaldi, Peter J

2018-01-01

Emphysema has considerable variability in its regional distribution. Craniocaudal emphysema distribution is an important predictor of the response to lung volume reduction. However, there is little consensus regarding how to define upper lobe-predominant and lower lobe-predominant emphysema subtypes. Consequently, the clinical and genetic associations with these subtypes are poorly characterized. We sought to identify subgroups characterized by upper-lobe or lower-lobe emphysema predominance and comparable amounts of total emphysema by analyzing data from 9,210 smokers without alpha-1-antitrypsin deficiency in the Genetic Epidemiology of COPD (COPDGene) cohort. CT densitometric emphysema was measured in each lung lobe. Random forest clustering was applied to lobar emphysema variables after regressing out the effects of total emphysema. Clusters were tested for association with clinical and imaging outcomes at baseline and at 5-year follow-up. Their associations with genetic variants were also compared. Three clusters were identified: minimal emphysema (n = 1,312), upper lobe-predominant emphysema (n = 905), and lower lobe-predominant emphysema (n = 796). Despite a similar amount of total emphysema, the lower-lobe group had more severe airflow obstruction at baseline and higher rates of metabolic syndrome compared with subjects with upper-lobe predominance. The group with upper-lobe predominance had greater 5-year progression of emphysema, gas trapping, and dyspnea. Differential associations with known COPD genetic risk variants were noted. Subgroups of smokers defined by upper-lobe or lower-lobe emphysema predominance exhibit different functional and radiological disease progression rates, and the upper-lobe predominant subtype shows evidence of association with known COPD genetic risk variants. These subgroups may be useful in the development of personalized treatments for COPD. Copyright © 2017 American College of Chest Physicians. Published by Elsevier Inc. All rights reserved.

Estimation of individual sennosides in plant materials and marketed formulations by an HPTLC method.

PubMed

Shah, S A; Ravishankara, M N; Nirmal, A; Shishoo, C J; Rathod, I S; Suhagia, B N

2000-04-01

Senna is a well-known drug, used in the Ayurvedic and Allopathic systems of medicine, and is a treatment for constipation. The purgative action of senna and its formulations is due to the presence of sennosides A and B. An HPTLC method has been developed for the determination of individual sennosides (A, B, C, D) without any derivatization in marketed formulations (three tablet formulations, two granule formulations and one liquid formulation) and plant materials (senna leaf and pod). The methanolic solution of a sample was applied on a pre-coated silica gel G60 F254 TLC plate (E. Merck.) and was developed using n-propanol : ethyl acetate : water : glacial acetic acid (3 : 3 : 2 : 0.1 v/v) as the mobile phase. The relative band speeds (Rf values) obtained were 0.35, 0.25, 0.61, 0.46 for sennosides A, B, C and D, respectively. The densitometric response was monitored at 366nm. Calibration curves were found to be linear in the concentration ranges 193-1356, 402-2817, 71-497 and 132-927 ng per spot for sennosides A, B, C, and D, respectively. The correlation coefficients were found to be 0.9978, 0.9987, 0.9939 and 0.9983 respectively for sennosides A, B, C and D. The result obtained with the HPTLC method for total sennoside content was compared with the results using the pharmacopoeial methods (spectrophotometric (British Pharmacopoeia) and spectrofluorimetric (United States Pharmacopeia) using the 'F' test). The results revealed no significant difference in the three different methods for estimation of total sennoside. The proposed HPTLC method was found to be simple, specific, precise, accurate and rapid. It can be used for routine quality control of sennosides or senna-containing formulations for individual sennosides.

Ameliorating effect of Celastrus paniculatus standardized extract and its fractions on 3-nitropropionic acid induced neuronal damage in rats: possible antioxidant mechanism.

PubMed

Malik, Jai; Karan, Maninder; Dogra, Rachna

2017-12-01

Celastrus paniculatus Wild. (Celasteraceae) (CP) is a well-known Ayurvedic 'Medhya Rasayana' (nervine tonic), used extensively as a neuro-protective and memory enhancer, and in different central nervous system disorders. To evaluate the effect of CP against 3-nitropropionic acid (3-NP) induced Huntington's disease (HD) like symptoms in Wistar male rats. The ethanol extract of CP seeds (CPEE), prepared by maceration, was standardized on the basis of linoleic acid content (6.42%) using thin layer chromatography densitometric analysis. Protective effect of CPEE (100 and 200 mg/kg) and its various fractions, viz., petroleum ether (40 mg/kg), ethyl acetate (2.5 mg/kg), n-butanol (7 mg/kg) and aqueous (18 mg/kg), administered orally for 20 days, against 3-NP (10 mg/kg, i.p. for 14 days) was assessed by their effect on body weight, locomotor activity, grip strength, gait pattern and cognitive dysfunction and biochemical parameters for oxidative damage in the striatum and cortex regions of the brain. CPEE (100 and 200 mg/kg) treated animals exhibited a significant (p < 0.05) improvement in behavioural and oxidative stress parameters in comparison to only 3-NP treated animals. Amongst various tested fractions of CPEE, aqueous fraction (AF) at 18 mg/kg exhibited maximum reversal of 3-NP induced behavioural and biochemical alterations, and was therefore also tested at 9 and 36 mg/kg. CPEE (100 mg/kg) and AF (36 mg/kg) exhibited maximum and significant (p < 0.05) attenuation of 3-NP induced alterations in comparison to 3-NP treated rats. CPEE has a protective action against 3-NP induced HD like symptoms due to its strong antioxidant effect.

Immunologic analysis of human breast cancer progesterone receptors. 1. Immunonaffinity purification of transformed receptors and production of monoclonal antibodies

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

Estes, P.A.; Suba, E.J.; Lawler-Heavner, J.

1987-09-22

A monoclonal antibody (MAb), designated PR-6, produced against chick oviduct progesterone receptors cross-reacts with the M/sub r/ 120,000 human B receptors. An immunomatrix prepared with PR-6 was used to purify progesterone receptors (PR) from T47D human breast cancer cells. Single-step immunoaffinity chromatography results in enrichment of B receptors (identified by immunoblot with PR-6 and by photoaffinity labeling with (/sup 3/H)promegestone) to a specific activity of 1915 pmol/mg of protein (or 23% purity) and with 27% yield. Purity and yields as judged by gel electrophoresis and densitometric scanning of the B protein were approximately 1.7-fold higher due to partial loss inmore » hormone binding activity at the elution step. B receptors purified under these conditions are transformed and biologically active. They were maintained as undergraded 120-kDa doublets and retained both hormone and DNA binding activities. These purified B receptors were used as immunogen for production of four monoclonal antibodies against human PR. Three of the MAbs, designated as B-30 (IgG/sub 1/), B-64 (IgG/sub 1/), and B-11 (IgM), are specific for B receptors. The fourth MAb, A/B-52 (IgG/sub 1/), reacts with both A and B receptors. The IgG MAbs are monospecific for human PR since they recognize and absorb native receptor-hormone complexes, displace the sedimentation of 4S receptors on salt containing sucrose gradients, and, by immunoblot assay of crude T47D cytosol, react only with receptor polypeptides. Although mice were injected with B receptors only, production of A/B-52 which recognized both A and B receptors provides evidence that these two proteins share regions of structural homology.« less

Mice maintain predominantly maternal Gαs expression throughout life in brown fat tissue (BAT), but not other tissues.

PubMed

Tafaj, Olta; Hann, Steven; Ayturk, Ugur; Warman, Matthew L; Jüppner, Harald

2017-10-01

The murine Gnas (human GNAS) locus gives rise to Gαs and different splice variants thereof. The Gαs promoter is not methylated thus allowing biallelic expression in most tissues. In contrast, the alternative first Gnas/GNAS exons and their promoters undergo parent specific methylation, which limits transcription to the non-methylated allele. Pseudohypoparathyroidism type Ia (PHP1A) or type Ib (PHP1B) are caused by heterozygous maternal GNAS mutations suggesting that little or no Gαs is derived in some tissues from the non-mutated paternal GNAS thereby causing hormonal resistance. Previous data had indicated that Gαs is mainly derived from the maternal Gnas allele in brown adipose tissue (BAT) of newborn mice, yet it is biallelically expressed in adult BAT. This suggested that paternal Gαs expression is regulated by an unknown factor(s) that varies considerably with age. To extend these findings, we now used a strain-specific SNP in Gnas exon 11 (rs13460569) for evaluation of parent-specific Gαs expression through the densitometric quantification of BanII-digested RT-PCR products and digital droplet PCR (ddPCR). At all investigated ages, Gαs transcripts were derived in BAT predominantly from the maternal Gnas allele, while kidney and liver showed largely biallelic Gαs expression. Only low or undetectable levels of other paternally Gnas-derived transcripts were observed, making it unlikely that these are involved in regulating paternal Gαs expression. Our findings suggest that a cis-acting factor could be implicated in reducing paternal Gαs expression in BAT and presumably in proximal renal tubules, thereby causing PTH-resistance if the maternal GNAS/Gnas allele is mutated. Copyright © 2017 Elsevier Inc. All rights reserved.

Computed Tomographic Measures of Pulmonary Vascular Morphology in Smokers and Their Clinical Implications

PubMed Central

Estépar, Raúl San José; Kinney, Gregory L.; Black-Shinn, Jennifer L.; Bowler, Russell P.; Kindlmann, Gordon L.; Ross, James C.; Kikinis, Ron; Han, MeiLan K.; Come, Carolyn E.; Diaz, Alejandro A.; Cho, Michael H.; Hersh, Craig P.; Schroeder, Joyce D.; Reilly, John J.; Lynch, David A.; Crapo, James D.; Wells, J. Michael; Dransfield, Mark T.; Hokanson, John E.

2013-01-01

Rationale: Angiographic investigation suggests that pulmonary vascular remodeling in smokers is characterized by distal pruning of the blood vessels. Objectives: Using volumetric computed tomography scans of the chest we sought to quantitatively evaluate this process and assess its clinical associations. Methods: Pulmonary vessels were automatically identified, segmented, and measured. Total blood vessel volume (TBV) and the aggregate vessel volume for vessels less than 5 mm2 (BV5) were calculated for all lobes. The lobe-specific BV5 measures were normalized to the TBV of that lobe and the nonvascular tissue volume (BV5/TissueV) to calculate lobe-specific BV5/TBV and BV5/TissueV ratios. Densitometric measures of emphysema were obtained using a Hounsfield unit threshold of −950 (%LAA-950). Measures of chronic obstructive pulmonary disease severity included single breath measures of diffusing capacity of carbon monoxide, oxygen saturation, the 6-minute-walk distance, St George’s Respiratory Questionnaire total score (SGRQ), and the body mass index, airflow obstruction, dyspnea, and exercise capacity (BODE) index. Measurements and Main Results: The %LAA-950 was inversely related to all calculated vascular ratios. In multivariate models including age, sex, and %LAA-950, lobe-specific measurements of BV5/TBV were directly related to resting oxygen saturation and inversely associated with both the SGRQ and BODE scores. In similar multivariate adjustment lobe-specific BV5/TissueV ratios were inversely related to resting oxygen saturation, diffusing capacity of carbon monoxide, 6-minute-walk distance, and directly related to the SGRQ and BODE. Conclusions: Smoking-related chronic obstructive pulmonary disease is characterized by distal pruning of the small blood vessels (<5 mm2) and loss of tissue in excess of the vasculature. The magnitude of these changes predicts the clinical severity of disease. PMID:23656466

Combining simplicity with cost-effectiveness: Investigation of potential counterfeit of proton pump inhibitors through simulated formulations using thin-layer chromatography.

PubMed

Bhatt, Nejal M; Chavada, Vijay D; Sanyal, Mallika; Shrivastav, Pranav S

2016-11-18

A simple, accurate and precise high-performance thin-layer chromatographic method has been developed and validated for the analysis of proton pump inhibitors (PPIs) and their co-formulated drugs, available as binary combination. Planar chromatographic separation was achieved using a single mobile phase comprising of toluene: iso-propranol: acetone: ammonia 5.0:2.3:2.5:0.2 (v/v/v/v) for the analysis of 14 analytes on aluminium-backed layer of silica gel 60 FG 254 . Densitometric determination of the separated spots was done at 290nm. The method was validated according to ICH guidelines for linearity, precision and accuracy, sensitivity, specificity and robustness. The method showed good linear response for the selected drugs as indicated by the high values of correlation coefficients (≥0.9993). The limit of detection and limit of quantiation were in the range of 6.9-159.2ng/band and 20.8-478.1ng/band respectively for all the analytes. The optimized conditions afforded adequate resolution of each PPI from their co-formulated drugs and provided unambiguous identification of the co-formulated drugs from their homologous retardation factors (hR f ). The only limitation of the method was the inability to separate two PPIs, rabeprazole and lansoprazole from each other. Nevertheless, it is proposed that peak spectra recording and comparison with standard drug spot can be a viable option for assignment of TLC spots. The method performance was assessed by analyzing different laboratory simulated mixtures and some marketed formulations of the selected drugs. The developed method was successfully used to investigate potential counterfeit of PPIs through a series of simulated formulations with good accuracy and precision. Copyright © 2016 Elsevier B.V. All rights reserved.

Multi-enzyme inhibition assay for the detection of insecticidal organophosphates and carbamates by high-performance thin-layer chromatography applied to determine enzyme inhibition factors and residues in juice and water samples.

PubMed

Akkad, Rami; Schwack, Wolfgang

2010-05-15

Esterase inhibition assays provide an effect-directed tool of rapid screening for inhibitors in environmental and food samples. According to a multi-enzyme microtiter-plate assay, rabbit liver esterase (RLE), Bacillus subtilis esterase (BS2), and cutinase from Fusarium solani pisi (CUT) were used for the detection of 21 organophosphorus and carbamate pesticides by high-performance thin-layer chromatography-enzyme inhibition assays (HPTLC-EI). Staining was performed with Fast Blue Salt B coupling to alpha-naphthol enzymatically released from the respective acetate used as substrate. Quantitative analysis was achieved by densitometric evaluation at 533 nm. Enzyme inhibition factors derived from HPTLC-EI were calculated from the slopes of the linear calibration curves, which allowed comparisons to published inhibition constants and well correlated to sensitivity parameters. Limits of detection ranged from a few pg/zone for organophosphates as strongest inhibitors to a few ng/zone for most carbamates, when RLE and BS2 were used. Without oxidation, chlorpyrifos and parathion were directly detectable at approximately 60 and 14 ng/zone, respectively. As the enzyme of lowest sensitivity, CUT was able to detect insecticides of high and low inhibitory power from the ng to microg range per zone. Due to high selectivity of enzyme inhibition, oxon impurities of thionophosphate standards were strongly detected, although only present in low traces. The exemplary application of HPTLC-EI (RLE) to apple juice and drinking water samples spiked with paraoxon (0.001 mg/L), parathion (0.05 mg/L) and chlorpyrifos (0.5mg/L) resulted in mean recoveries between 71 and 112% with standard deviations of 2.0-18.3%. Copyright (c) 2009 Elsevier B.V. All rights reserved.

Stability-indicating assay of repaglinide in bulk and optimized nanoemulsion by validated high performance thin layer chromatography technique.

PubMed

Akhtar, Juber; Fareed, Sheeba; Aqil, Mohd

2013-07-01

A sensitive, selective, precise and stability-indicating high-performance thin-layer chromatographic (HPTLC) method for analysis of repaglinide both as a bulk drug and in nanoemulsion formulation was developed and validated. The method employed TLC aluminum plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of chloroform/methanol/ammonia/glacial acetic acid (7.5:1.5:0.9:0.1, v/v/v/v). This system was found to give compact spots for repaglinide (R f value of 0.38 ± 0.02). Repaglinide was subjected to acid and alkali hydrolysis, oxidation, photodegradation and dry heat treatment. Also, the degraded products were well separated from the pure drug. Densitometric analysis of repaglinide was carried out in the absorbance mode at 240 nm. The linear regression data for the calibration plots showed good linear relationship with r (2)= 0.998 ± 0.032 in the concentration range of 50-800 ng. The method was validated for precision, accuracy as recovery, robustness and specificity. The limits of detection and quantitation were 0.023 and 0.069 ng per spot, respectively. The drug undergoes degradation under acidic and basic conditions, oxidation and dry heat treatment. All the peaks of the degraded product were resolved from the standard drug with significantly different R f values. Statistical analysis proves that the method is reproducible and selective for the estimation of the said drug. As the method could effectively separate the drug from its degradation products, it can be employed as a stability-indicating one. Moreover, the proposed HPTLC method was utilized to investigate the degradation kinetics in 1M NaOH.

Effect of transportation stress on heat shock protein 70 concentration and mRNA expression in heart and kidney tissues and serum enzyme activities and hormone concentrations of pigs.

PubMed

Yu, Hong; Bao, En-Dong; Zhao, Ru-Qian; Lv, Qiong-Xia

2007-11-01

To determine the enzymatic and hormonal responses, heat shock protein 70 (Hsp70) production, and Hsp70 mRNA expression in heart and kidney tissues of transport-stressed pigs. 24 pigs (mean weight, 20 +/- 1 kg). Pigs were randomly placed into groups of 12 each. One group was transported for 2 hours. The other group was kept under normal conditions and used as control pigs. Sera were used to detect triiodothyronine, thyroxine, and cortisol concentrations and alanine aminotransferase, aspartate aminotransferase, and creatine kinase activities. The heart and kidneys of anesthetized pigs were harvested and frozen in liquid nitrogen for quantification of Hsp70 and Hsp70 mRNA. No significant differences were detected in serum alanine aminotransferase activity and triiodothyronine and cortisol concentrations between groups; however, the serum creatine kinase and aspartate aminotransferase activities and thyroxine concentrations were higher in transported pigs. Densitometric readings of western blots revealed that the amount of Hsp70 in heart and kidney tissues was significantly higher in transported pigs, compared with control pigs. Results of fluorescence quantitative real-time PCR assay revealed that the Hsp70 mRNA transcription in heart tissue, but not kidney tissue, was significantly higher in transported pigs, compared with control pigs. Transportation imposed a severe stress on pigs that was manifested as increased serum activities of aspartate aminotransferase and creatine kinase and increased amounts of Hsp70 and Hsp70 mRNA expression in heart and kidney tissues. Changes in serum enzyme activities were related to the tissue damage of transport-stressed pigs.

Allergen cross-reactivity between Pityrosporum orbiculare and Candida albicans.

PubMed

Huang, X; Johansson, S G; Zargari, A; Nordvall, S L

1995-08-01

Pityrosporum orbiculare and Candida albicans extracts were separated by SDS-PAGE, and IgE binding was detected by immunoblotting with 21 patient sera that were RAST positive to both yeasts. Cross-wise inhibition was performed of IgE binding of a serum pool containing IgE antibodies to both yeasts. The pool was mixed with serial dilutions of P. orbiculare or C. albicans extracts, and incubated with strips containing separated allergen. IgE binding was quantified by densitometric scanning and percent inhibition was calculated as well as the respective ratios between required extract concentration for 50% inhibition in heterologous compared to homologous inhibition for each component (inhibition ratio). Ten components of P. orbiculare were detected by more than 60% of the sera. IgE binding to C. albicans was weak, and only to four bands was IgE binding detected by more than 30% of the sera. The most important C. albicans allergen was a 48-kDa band, to which IgE of half of the patient sera bound. There was little inhibition of IgE binding to P. orbiculare with C. albicans. Thus, all but three components exhibited an inhibition ratio higher than 100. The inhibition ratio of the 48-kDa C. albicans compound was 50, thus indicating some degree of cross-reactivity. Significant cross-reactivity was shown by C. albicans compounds of 18, 24, 26, 34, and 38 kDa, the inhibition ratios of which were less than 10. There was some degree of cross-reactivity between apparent protein allergens of the two yeasts, but IgE antibodies to C. albicans do not merely reflect sensitization to P. orbiculare.

Prevalence and impact of bronchiectasis in alpha1-antitrypsin deficiency.

PubMed

Parr, David G; Guest, Peter G; Reynolds, John H; Dowson, Lee J; Stockley, Robert A

2007-12-15

alpha(1)-Antitrypsin (AAT) deficiency is associated with increased risk of chronic obstructive pulmonary disease (COPD), in particular emphysema, but airway disease is less well described. To assess the prevalence of airways disease in subjects with AAT deficiency and to identify the relationship between radiological airway abnormalities and clinical phenotype. We characterized the computed tomographic phenotype of 74 subjects (PiZ), using visual scoring of airway disease and densitometric assessment of emphysema. Computed tomographic measurements were related to physiology, health status (St. George's Respiratory Questionnaire), and emphysema severity, and the relative impact of airway disease and emphysema severity on health status and airflow obstruction was compared by stepwise regression. Bronchiectatic changes were seen in 70 subjects, and a subgroup with a bronchiectasis-predominant phenotype was identified. Clinically significant bronchiectasis (radiologic bronchiectasis in 4 or more bronchopulmonary segments together with symptoms of regular sputum production) occurred in 20 subjects (27%). AAT-deficient index cases had higher airway disease scores (P < 0.05), more severe emphysema (P < 0.001), and greater impairment of physiology (P < 0.001) and health status (P < 0.05) than nonindex cases. Airway disease scores correlated with health status, and bronchial wall thickening correlated with FEV(1). Regression analysis indicated that emphysema severity had the strongest associations for health status (r = 0.505, P < 0.001) and FEV(1) (r = 0.699, P < 0.001), but the addition of airway disease score improved the regression models (r = 0.596, P = 0.002 and r = 0.783, P < 0.001, respectively). Emphysema is the predominant component of COPD in AAT deficiency, but the prevalence and impact of airway disease are greater than currently recognized. Consequently, future therapeutic strategies in AAT deficiency should also target this component of COPD.

Sex-related differences of bone properties of pelvic limb and bone metabolism indices in 14-month-old ostriches (Struthio camelus).

PubMed

Krupski, W; Tatara, M R; Charuta, A; Brodzki, A; Szpetnar, M; Jóźwik, A; Strzałkowska, N; Poławska, E; Łuszczewska-Sierakowska, I

2018-06-01

1. Sex-related differences of long pelvic limb bones and serum bone metabolism indices were evaluated in 14-month-old female (N = 7) and male (N = 7) ostriches of similar body weights. 2. Densitometric parameters of femur, tibia and tarsometatarsus were determined using quantitative computed tomography (volumetric bone mineral density, calcium hydroxyapatite density and mean volumetric bone mineral density) and dual energy X-ray absorptiometry (bone mineral density and bone mineral content) methods. Geometrical parameters such as cortical bone area, cross-sectional area, second moment of inertia, mean relative wall thickness and cortical index were determined in the midshaft of bones. Mechanical properties of bones (maximum elastic strength and ultimate strength) were evaluated using three-point bending test. Serum concentrations of free amino acids, osteocalcin, N-terminal propeptide of type I procollagen, C-terminal telopeptides of type II collagen and total antioxidative capacity were also determined. 3. Bone weight and relative bone weight of all bones were significantly higher in males than in females. Significantly lower values of trabecular bone mineral density and calcium hydroxyapatite density were found in the trabecular bone of tibia in males. The highest number of the sex-related differences was observed in the tarsometatarsus where bone length, bone mineral content, cortical bone area, cross-sectional area and ultimate strength were higher in males. Serum concentrations of taurine, hydroxyproline, valine and isoleucine were significantly higher in males. 4. Higher loading of the tarsometatarsus in comparison to femur and tibia may be an important factor interacting with sex hormones in regulation of bone formation and mineralisation processes. Sex-related differences of bone properties were associated with increased serum concentration of selected amino acids in males.

NADP+ Binding to the Regulatory Subunit of Methionine Adenosyltransferase II Increases Intersubunit Binding Affinity in the Hetero-Trimer

PubMed Central

Ortega, Rebeca; Martínez-Júlvez, Marta; Revilla-Guarinos, Ainhoa; Pérez-Pertejo, Yolanda; Velázquez-Campoy, Adrián; Sanz-Aparicio, Julia; Pajares, María A.

2012-01-01

Mammalian methionine adenosyltransferase II (MAT II) is the only hetero-oligomer in this family of enzymes that synthesize S-adenosylmethionine using methionine and ATP as substrates. Binding of regulatory β subunits and catalytic α2 dimers is known to increase the affinity for methionine, although scarce additional information about this interaction is available. This work reports the use of recombinant α2 and β subunits to produce oligomers showing kinetic parameters comparable to MAT II purified from several tissues. According to isothermal titration calorimetry data and densitometric scanning of the stained hetero-oligomer bands on denatured gels, the composition of these oligomers is that of a hetero-trimer with α2 dimers associated to single β subunits. Additionally, the regulatory subunit is able to bind NADP+ with a 1∶1 stoichiometry, the cofactor enhancing β to α2-dimer binding affinity. Mutants lacking residues involved in NADP+ binding and N-terminal truncations of the β subunit were able to oligomerize with α2-dimers, although the kinetic properties appeared altered. These data together suggest a role for both parts of the sequence in the regulatory role exerted by the β subunit on catalysis. Moreover, preparation of a structural model for the hetero-oligomer, using the available crystal data, allowed prediction of the regions involved in β to α2-dimer interaction. Finally, the implications that the presence of different N-terminals in the β subunit could have on MAT II behavior are discussed in light of the recent identification of several splicing forms of this subunit in hepatoma cells. PMID:23189196

Identification of structural and secretory lectin-binding glycoproteins of normal and cancerous human prostate.

PubMed

Lad, P M; Cooper, J F; Learn, D B; Olson, C V

1984-12-07

We have utilized the technique of lectin-loading of SDS gels with iodinated concanavalin A and wheat germ agglutinin to identify glycoproteins in prostatic and seminal fluids as well as in prostate tissue fractions. The following subunits which bound both lectins were detected: (a) 50, 43 and 38 kDa subunits common to prostatic and seminal fluids, and an additional 55 kDa subunit which predominates only in prostatic fluid; (b) 78, 55, 50 and 43 kDa subunits in prostatic tissue cytosol and (c) 195, 170, 135, 116 and 95 kDa subunits present in the particulate fractions of prostatic tissue. Immunoblotting using specific rabbit antibodies revealed the 50 kDa band to be prostatic acid phosphatase and the 38 kDa band to be prostate-specific antigen. Interestingly, antibodies directed toward prostatic acid phosphatase were found to cross-react with the 43 kDa band. Fractionation on sucrose gradients showed that several of these particulate glycoproteins were associated with a vesicle fraction enriched in adenylate cyclase activity, implying that they are plasma membrane glycoproteins. Comparison of soluble and particulate fractions of normal and cancerous tissue homogenates was made by densitometric scanning of autoradiograms of lectin-loaded gels. Similar relative intensities of lectin-binding were obtained for corresponding proteins in normal and cancerous tissue fractions. Also, immunoblotting showed no differences in prostatic acid phosphatase or prostate-specific antigen between normal and cancerous soluble homogenate fractions. Our results suggest that major lectin-binding proteins are conserved in the transition from normal to cancerous tissue. These results may be useful in developing a multiple-marker profile of metastatic prostate cancer and for the design of imaging agents, such as monoclonal antibodies, to prominent soluble and particulate prostate glycoproteins.

Cigarette smoking and alveolar bone in young adults: a study using digitized radiographs.

PubMed

Rosa, Guillermo M; Lucas, Gabriela Q; Lucas, Oscar N

2008-02-01

Evidence indicates that cigarette smoking is one of the most significant risk factors for periodontal diseases; however, there have been few radiographic prospective studies of alveolar bone in young populations. The purpose of this study was to evaluate the effect of smoking on alveolar bone in young adults. Eighty-one dental students (mean age: 20.5 years), considered not to have periodontitis according to clinical criteria, participated in this study. Forty-two subjects were smokers (mean consumption was 14.1 cigarettes/day for > or =2 years), and 39 subjects had never smoked. A parallel-arm prospective design was used. All subjects took part in a dental hygiene program (DHP) that included oral hygiene instructions, mechanical debridement, and polishing. The following clinical variables were measured before and after the DHP: plaque index (PI), gingival crevicular fluid (GCF) flow rate, gingival index (GI), probing depth, and clinical attachment level (CAL). Standardized posterior vertical bitewing radiographs were taken and digitized preexperimentally and on days 180, 365, and 545. The following analyses were performed: bone height measurement (BHM), computer-assisted densitometric image analysis (CADIA), and qualitative analysis of digital subtraction radiography (DSR). Repeated-measures multiple-way analysis of variance (ANOVA) was performed between the groups, and one-way ANOVA was performed within the groups. The mean PI and GI were significantly greater in the smokers (P <0.01). The mean GCF flow rate was significantly lower in the smokers (P <0.01). CAL and the number of sites with recession were significantly greater in the smokers (P <0.001). The BHM indicated a significantly lower mean alveolar bone height in the smokers (P <0.01). The smokers showed significantly lower CADIA values, which indicated a lower bone density on days 0 (P <0.05), 180, 365, and 545 (P <0.01). CADIA values decreased during the study in the smokers, with significant differences on day 545 (P <0.05). The smokers had a significantly higher mean percentage of sites that had decreased density, as assessed by DSR (P <0.001). In the smokers, the mean percentage of sites with decreased density, as assessed by DSR, had increased significantly by days 365 (P <0.05) and 545 (P <0.01). Smoking produces an adverse effect on clinical periodontal variables and alveolar bone height and density, acting as a potential risk factor for alveolar bone loss, even at an early age with low tobacco consumption. It is very important to inform young smokers about the risk of this habit in relation to periodontal health.

Pro-oxidant status and matrix metalloproteinases in apical lesions and gingival crevicular fluid as potential biomarkers for asymptomatic apical periodontitis and endodontic treatment response

PubMed Central

2012-01-01

Background Oxidative stress and matrix metalloproteinases -9 and -2 are involved in periodontal breakdown, whereas gingival crevicular fluid has been reported to reflect apical status. The aim of this study was to characterize oxidant balance and activity levels of MMP -2 and -9 in apical lesions and healthy periodontal ligament; and second, to determine whether potential changes in oxidant balance were reflected in gingival crevicular fluid from asymptomatic apical periodontitis (AAP)-affected teeth at baseline and after endodontic treatment. Methods Patients with clinical diagnosis of AAP and healthy volunteers having indication of tooth extraction were recruited. Apical lesions and healthy periodontal ligaments, respectively, were homogenized or processed to obtain histological tissue sections. Matrix metalloproteinase -9 and -2 levels and/or activity were analyzed by Immunowestern blot, zymography and consecutive densitometric analysis, and their tissue localization was confirmed by immunohistochemistry. A second group of patients with AAP and indication of endodontic treatment was recruited. Gingival crevicular fluid was extracted from AAP-affected teeth at baseline, after endodontic treatment and healthy contralateral teeth. Total oxidant and antioxidant status were determined in homogenized tissue and GCF samples. Statistical analysis was performed using STATA v10 software with unpaired t test, Mann-Whitney test and Spearman's correlation. Results Activity of MMP-2 and MMP-9 along with oxidant status were higher in apical lesions (p < 0.05). Total oxidant status correlated positively with matrix metalloproteinase-2 and lesion size (p < 0.05). Gingival crevicular fluid showed significantly lower levels of total antioxidant status in diseased teeth at baseline compared to controls and endodontically-treated groups. Conclusions Apical lesions display an oxidant imbalance along with increased activity of matrix metalloproteinase-2 and -9 and might contribute to AAP progression. Oxidant imbalance can also be reflected in GCF from AAP-affected teeth and was restored to normal levels after conservative endodontic treatment. These mediators might be useful as potential biomarkers for chair-side complementary diagnostic of apical status in GCF. PMID:22436166

Pro-oxidant status and matrix metalloproteinases in apical lesions and gingival crevicular fluid as potential biomarkers for asymptomatic apical periodontitis and endodontic treatment response.

PubMed

Dezerega, Andrea; Madrid, Sonia; Mundi, Verónica; Valenzuela, María A; Garrido, Mauricio; Paredes, Rodolfo; García-Sesnich, Jocelyn; Ortega, Ana V; Gamonal, Jorge; Hernández, Marcela

2012-03-21

Oxidative stress and matrix metalloproteinases -9 and -2 are involved in periodontal breakdown, whereas gingival crevicular fluid has been reported to reflect apical status. The aim of this study was to characterize oxidant balance and activity levels of MMP -2 and -9 in apical lesions and healthy periodontal ligament; and second, to determine whether potential changes in oxidant balance were reflected in gingival crevicular fluid from asymptomatic apical periodontitis (AAP)-affected teeth at baseline and after endodontic treatment. Patients with clinical diagnosis of AAP and healthy volunteers having indication of tooth extraction were recruited. Apical lesions and healthy periodontal ligaments, respectively, were homogenized or processed to obtain histological tissue sections. Matrix metalloproteinase -9 and -2 levels and/or activity were analyzed by Immunowestern blot, zymography and consecutive densitometric analysis, and their tissue localization was confirmed by immunohistochemistry. A second group of patients with AAP and indication of endodontic treatment was recruited. Gingival crevicular fluid was extracted from AAP-affected teeth at baseline, after endodontic treatment and healthy contralateral teeth. Total oxidant and antioxidant status were determined in homogenized tissue and GCF samples. Statistical analysis was performed using STATA v10 software with unpaired t test, Mann-Whitney test and Spearman's correlation. Activity of MMP-2 and MMP-9 along with oxidant status were higher in apical lesions (p < 0.05). Total oxidant status correlated positively with matrix metalloproteinase-2 and lesion size (p < 0.05). Gingival crevicular fluid showed significantly lower levels of total antioxidant status in diseased teeth at baseline compared to controls and endodontically-treated groups. Apical lesions display an oxidant imbalance along with increased activity of matrix metalloproteinase-2 and -9 and might contribute to AAP progression. Oxidant imbalance can also be reflected in GCF from AAP-affected teeth and was restored to normal levels after conservative endodontic treatment. These mediators might be useful as potential biomarkers for chair-side complementary diagnostic of apical status in GCF.

Abnormal distal renal tubular acidification in patients with low bone mass: prevalence and impact of alkali treatment.

PubMed

Sromicki, Jerzy Jan; Hess, Bernhard

2017-06-01

Chronic acid retention is known to promote bone dissolution. In this study, 23 % of patients with osteopenia/osteoporosis were diagnosed with abnormal distal renal tubular acidification (dRTA), a kidney dysfunction leading to chronic acid retention. Treating those patients with alkali-therapy shows improvement in bone density. To evaluate the prevalence of abnormal distal renal tubular acidification in patients with low bone mass (LBM) and the impact of additional alkali treatment on bone density in patients with concomitant LBM and dRTA,183 patients referred for metabolic evaluation of densitometrically proven low bone mass were screened for abnormal distal renal tubular acidification between 2006 and 2013. In all LBM urine pH (U-pH) was measured in the 2nd morning urines after 12 h of fasting. If U-pH was ≥5.80, LBM underwent a 1-day ammonium chloride loading, and U-pH was remeasured the next morning. If U-pH after acid loading did not drop below 5.45, patients were diagnosed with abnormal distal renal tubular acidification. Normal values were obtained from 21 healthy controls. All LBM with dRTA were recommended alkali citrate in addition to conventional therapy of LBM, and follow-up DXAs were obtained until 2014. 85 LBM underwent NH 4 Cl loading. 42 LBM patients were diagnosed with incomplete dRTA (idRTA; prevalence 23.0 %). During follow-up (1.6-8 years) of idRTA-LBM patients, subjects adhering to alkali treatment tended to improve BMD at all sites measured, whereas BMD of non-adherent idRTA patients worsened/remained unchanged. (1) About one out of four patients with osteopenia/osteoporosis has idRTA. (2) Upon NH 4 Cl loading, idRTA patients do not lower urine pH normally, but show signs of increased acid-buffering by bone dissolution. (3) In idRTA patients with low bone mass on conventional therapy, additional long-term alkali treatment improves bone mass at lumbar spine and potentially at other bone sites. (4) All patients with low bone mass undergoing metabolic evaluation should be screened for idRTA.

High SMAD7 and p-SMAD2,3 expression is associated with environmental enteropathy in children.

PubMed

Syed, Sana; Dinallo, Vincenzo; Iqbal, Najeeha T; Di Iorio, Laura; Di Fusco, Davide; Guleria, Shan; Amadi, Beatrice C; Sadiq, Kamran; Moskaluk, Christopher; Ali, S Asad; Kelly, Paul; Monteleone, Giovanni

2018-02-01

Enteropathies such as Crohn's disease are associated with enteric inflammation characterized by impaired TGF-β signaling, decreased expression of phosphorylated (p)-SMAD2,3 and increased expression of SMAD7 (an inhibitor of SMAD3 phosphorylation). Environmental enteropathy (EE) is an acquired inflammatory disease of the small intestine (SI), which is associated with linear growth disruption, cognitive deficits, and reduced oral vaccine responsiveness in children <5 y in resource-poor countries. We aimed to characterize EE inflammatory pathways by determining SMAD7 and p-SMAD2,3 levels (using Western blotting) in EE duodenal biopsies (N = 19 children, 7 from Pakistan, 12 from Zambia) and comparing these with healthy controls (Ctl) and celiac disease (CD) patients from Italy. Densitometric analysis of immunoblots showed that EE SI biopsies expressed higher levels of both SMAD7 (mean±SD in arbitrary units [a.u.], Ctl = 0.47±0.20 a.u., EE = 1.13±0.25 a.u., p-value = 0.03) and p-SMAD2,3 (mean±SD, Ctl = 0.38±0.14 a.u., EE = 0.60±0.10 a.u., p-value = 0.03). Immunohistochemistry showed that, in EE, SMAD7 is expressed in both the epithelium and in mononuclear cells of the lamina propria (LP). In contrast, p-SMAD3 in EE is expressed much more prominently in epithelial cells than in the LP. The high SMAD7 immunoreactivity and lack of p-SMAD3 expression in the LP suggests defective TGF-β signaling in the LP in EE similar to a previously reported SMAD7-mediated inflammatory pathway in refractory CD and Crohn's disease. However, Western blot densitometry showed elevated p-SMAD2,3 levels in EE, possibly suggesting a different inflammatory pathway than Crohn's disease but more likely reflecting cumulative protein expression from across all compartments of the mucosa as opposed to the LP alone. Further studies are needed to substantiate these preliminary results and to illustrate the relationship between SMAD proteins, TGF-β signaling, and inflammatory cytokine production, all of which may be potential therapeutic targets.

Quantitation of Human Cytochrome P450 2D6 Protein with Immunoblot and Mass Spectrometry Analysis

PubMed Central

Yu, Ai-Ming; Qu, Jun; Felmlee, Melanie A.; Cao, Jin; Jiang, Xi-Ling

2009-01-01

Accurate quantification of cytochrome P450 (P450) protein contents is essential for reliable assessment of drug safety, including the prediction of in vivo clearance from in vitro metabolism data, which may be hampered by the use of uncharacterized standards and existence of unknown allelic isozymes. Therefore, this study aimed to delineate the variability in absolute quantification of polymorphic CYP2D6 drug-metabolizing enzyme and compare immunoblot and nano liquid chromatography coupled to mass spectrometry (nano-LC/MS) methods in identification and relative quantification of CYP2D6.1 and CYP2D6.2 allelic isozymes. Holoprotein content of in-house purified CYP2D6 isozymes was determined according to carbon monoxide difference spectrum, and total protein was quantified with bicinchoninic acid protein assay. Holoprotein/total CYP2D6 protein ratio was markedly higher for purified CYP2D6.1 (71.0%) than that calculated for CYP2D6.1 Supersomes (35.5%), resulting in distinct linear calibration range (0.05–0.50 versus 0.025–0.25 pmol) that was determined by densitometric analysis of immunoblot bands. Likewise, purified CYP2D6.2 and CYP2D6.10 and the CYP2D6.10 Supersomes all showed different holoprotein/total CYP2D6 protein ratios and distinct immunoblot linear calibration ranges. In contrast to immunoblot, nano-LC/MS readily distinguished CYP2D6.2 (R296C and S486T) from CYP2D6.1 by isoform-specific proteolytic peptides that contain the altered amino acid residues. In addition, relative quantitation of the two allelic isozymes was successfully achieved with label-free protein quantification, consistent with the nominated ratio. Because immunoblot and nano-LC/MS analyses measure total P450 protein (holoprotein and apoprotein) in a sample, complete understanding of holoprotein and apoprotein contents in P450 standards is desired toward reliable quantification. Our data also suggest that nano-LC/MS not only facilitates P450 quantitation but also provides genotypic information. PMID:18832475

Influence of obesity on vertebral fracture prevalence and vitamin D status in postmenopausal women.

PubMed

El Maghraoui, A; Sadni, S; El Maataoui, A; Majjad, A; Rezqi, A; Ouzzif, Z; Mounach, A

2015-01-01

It is well established that weight is an important determinant of bone health. Whereas obesity is associated with increased mortality and morbidity from diabetes and cardiovascular diseases, high body weight is widely believed to be associated to hypovitaminosis D and protective against the development of osteoporosis and fracture risk. The objective of the study was to evaluate the effect of BMI on vitamin D status and on densitometric vertebral fractures (VFs) in a large series of asymptomatic women aged over 50 who had a VFA examination during their bone mineral density (BMD) testing. We enrolled 429 postmenopausal women (mean age, weight and BMI of 59.5 ± 8.3 (50 to 83) years, 75.8 ± 13.3 (35 to 165) kgs and 29.9 ± 5.2 (14.6 to 50.8) kg/m(2), respectively. Lateral VFA images and scans of the lumbar spine and proximal femur were obtained using a Lunar Prodigy densitometer. VFs were defined using the Genant semiquantitative (SQ) approach. Clinical risk factors of osteoporosis were collected and 25-hydroxivitamin D was measured using electrochimiluminescence (Roche). Prevalence of osteoporosis and hypovitaminosis D (<20 ng/ml) was 21.0 % and 78.1 % respectively. VFs grade 2/3were identified in 76 (17.7 %). Comparison between women according to their BMI showed that obese women had a higher BMD and less proportion of women with osteoporosis and VFs grade 2/3 than lean and overweight women. The prevalence of VFs globally increased with age and as BMI and BMD declined. Stepwise regression analysis showed that the presence of osteoporosis was independently related to BMI and history of fractures while the presence of grade 2/3 VFs was independently related to age, hypovitaminosis D and years of menopause. Obese women had a higher BMD and lower prevalence of VFs. VFs were significantly related to age, hypovitaminosis D and years since menopause. However, among obese women, prevalence of VFs was increased in osteoporotic women.

Simultaneous Quantification of Forskolin and Iso-Forskolin in Coleus forskohlii (Wild.) Briq. and Identification of Elite Chemotype, Collected from Eastern Ghats (India).

PubMed

Shukla, Pushpendra Kumar; Misra, Ankita; Kumar, Manish; Jaichand; Singh, Kuldeep; Akhtar, Juber; Srivastava, Sharad; Agrawal, Pawan K; Singh Rawat, Ajay K

2018-01-01

Coleus forskohlii is a well-known industrially important medicinal plant, for its high forskolin content. A simple, selective, and sensitive high-performance thin layer chromatography (HPTLC) method was developed and validated for simultaneous quantification of forskolin and iso-forskolin in C. forskohlii germplasm collected from the Eastern Ghats, India. Chromatographic separation of the targeted marker(s) was obtained on precoated silica plates using toluene: ethyl acetate: methanol (90:30:0.5, v/v/v) as the mobile phase. Densitometric quantification of forskolin and iso-forskolin was carried out at 545 nm. Forskolin and iso-forskolin were identified by comparing the ultraviolet spectra of standard and sample track at R f of 0.64 ± 0.02 and 0.36 ± 0.01, after derivatization with anisaldehyde sulfuric acid reagent. The linearity of both the analytes was obtained in the range of 300-1200 ng/spot with the regression coefficient ( R 2 ) of 0.991 and 0.986. Recovery of analyte (s) at three levels, namely, 100, 150, and 200 ng/spot was found to be 100.46% ± 0.29%, 99.64% ± 0.33%, 100.02% ± 0.76% and 99.76% ± 0.62%, 99.56% ± 0.35%, 100.02% ± 0.22%, respectively, for forskolin and iso-forskolin. The content of forskolin and iso-forskolin varies from 0.046% to 0.187% and 0.002% to 0.077%, respectively (dry weight basis), the maximum content of both the markers was found in NBC-31, from Thakurwada, Maharashtra. The developed HPTLC method was linear, accurate, and reliable as per the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The study aids in the identification of elite chemotype for commercial prospection of industrially viable medicinal crop. 12 Samples are collected from different locations of the eastern ghat regionsQuantification of two major marker forskolin and iso forskolinThe maximum content of both the markers was found in NBC -31, from Thakurwada, MaharashtraIdentification of elite chemotype of collected samples may be useful for commercial prospection in industries.

Upper and lower limbs composition: a comparison between anthropometry and dual-energy X-ray absorptiometry in healthy people.

PubMed

Diano, Danila; Ponti, Federico; Guerri, Sara; Mercatelli, Daniele; Amadori, Michele; Aparisi Gómez, Maria Pilar; Battista, Giuseppe; Guglielmi, Giuseppe; Bazzocchi, Alberto

2017-09-18

The detection of changes in lean mass (LM) distribution can help to prevent disability. This study assessed the degree of association between anthropometric measurements and dual-energy X-ray absorptiometry (DXA) body composition (BC) parameters of the upper and lower limbs in a healthy general population and collected DXA age- and sex-specific values of BC that can be useful to build a reference standard. The primary aim of this study was to investigate the reliability of some widely available anthropometric measurements in the assessment of body composition (BC) at the limbs, especially in terms of muscle mass, in a large sample of healthy subjects of different age bands and sex, using fat mass (FM) and lean mass (LM) parameters derived by dual-energy X-ray absorptiometry (DXA) as the gold standard. The secondary aim was to collect DXA age- and sex-specific values of BC of left and right limbs (upper and lower) in a healthy Italian population to be used as reference standards. Two hundred fifty healthy volunteers were enrolled. Arm circumference (AC) and thigh circumference (ThC) were measured, and total and regional BC parameters were obtained by a whole-body DXA scan (Lunar iDXA, Madison, WI, USA; enCORE™ 2011 software version 13.6). FM/LM showed only fair correlation with AC and ThC in females (r = 0.649 and 0.532, respectively); in males and in the total population, the correlation was low (r = 0.360 or lower, and p non-statistically significant). AC and ThC were not well representative of arms LM in both genders (females r = 0.452, males r = 0.530) independently of age. In general, men of all age groups showed higher values of LM and lean mass index (LMI) in both total and segmental upper and lower limbs. In males, the maximum LM and LMI were achieved in the fifth decade in both upper and lower limbs and then started to decrease with aging. In females, no significant modification with aging was identified in LM and LMI. According to our results, anthropometry is not well representative of LM of arms in both genders, independently of age; therefore, a densitometric examination should be considered for a correct assessment of BC at limbs.

Osteoporosis: Are we measuring what we intend to measure? In search of the ideal bone strength study

NASA Astrophysics Data System (ADS)

de Riese, Cornelia

2006-02-01

In 1991 the World Health Organization (WHO) defined osteoporosis as a "loss of bone mass and micro architectural deterioration of the skeleton leading to increased risk of fracture." 1,2 Since microarchitecture can not be measured directly, a panel of the WHO recommended that the diagnosis be made according to a quantifiable surrogate marker, calcium mineral, in bone. Subsequently in 1994, the definition focused on the actual bone "density," giving densitometric technology a central place in establishing the diagnosis of osteoporosis. 3,4 But soon it became obvious that there was only limited correlation between bone mineral density (BMD) and actual occurrence of fractures and that decreases in bone mass account for only about 50% of the deterioration of bone strength with aging. In other words only about 60% of bone strength is related to BMD. 5 Recent developments in bone research have shown that bone mineral density in itself is not sufficient to accurately predict fracture risk. Bone is composed of inorganic calcium apatite crystals that mineralize an organic type I collagen matrix. The degree of mineralization, the properties of the collagen matrix, crystal size, trabecular orientation, special distribution of the different components and many more factors are all impacting bone strength. 6-14 Human cadaver studies have confirmed the correlation between bone density and bone. 26 strength. 5,15-20 Changes in cancellous bone morphology appear to lead to a disproportionate decrease in bone strength. 21-26 When postmenopausal women are stratified by age, obvious differences between BMD and actual fracture risk are observed. 24 Felsenberg eloquently summarizes what he calls the "Bone Quality Framework." In great detail he talks about the geometry and micro- architecture of bone and how the different components are related to functional stability. 27 Are our current testing modalities appropriately addressing these structural factors? Are we keeping in mind that in screening for osteoporosis the key variable is fragility, not bone density itself? All currently FDA approved and commercially available equipments for the evaluation of bone status claim that they - at least indirectly - assess the biological fracture risk. This review summarizes an extensive current literature research covering FDA approved as well as experimental devices for the evaluation of bone. The pros and cons of the different techniques are discussed in the context of diagnostic accuracies and practical implications.

Early postnatal soluble FGFR3 therapy prevents the atypical development of obesity in achondroplasia

PubMed Central

Sarrazy, Vincent; Dumas, Karine; Authier, Florence; Sore, Sophie; Tran, Albert; Gual, Philippe; Gennero, Isabelle; Salles, Jean-Pierre; Gouze, Elvire

2018-01-01

Background Achondroplasia is a rare genetic disease is characterized by abnormal bone development and early obesity. While the bone aspect of the disease has been thoroughly studied, early obesity affecting approximately 50% of them during childhood has been somewhat neglected. It nevertheless represents a major health problem in these patients, and is associated to life-threatening complications including increasing risk of cardiovascular pathologies. We have thus decided to study obesity in patients and to use the mouse model to evaluate if soluble FGFR3 therapy, an innovative treatment approach for achondroplasia, could also impact the development of this significant complication. Methods and findings To achieve this, we have first fully characterized the metabolic deregulations in these patients by conducting a longitudinal retrospective study, in children with achondroplasia Anthropometric, densitometric measures as well as several blood parameters were recorded and compared between three age groups ranging from [0–3], [4–8] and [9–18] years old. Our results show unexpected results with the development of an atypical obesity with preferential fat deposition in the abdomen that is remarkably not associated with classical complications of obesity such as diabetes or hypercholosterolemia. Because it is not associated with diabetes, the atypical obesity has not been studied in the past even though it is recognized as a real problem in these patients. These results were validated in a murine model of achondroplasia (Fgfr3ach/+) where similar visceral adiposity was observed. Unexpected alterations in glucose metabolism were highlighted during high-fat diet. Glucose, insulin or lipid levels remained low, without the development of diabetes. Very interestingly, in achondroplasia mice treated with soluble FGFR3 during the growth period (from D3 to D22), the development of these metabolic deregulations was prevented in adult animals (between 4 and 14 weeks of age). The lean-over-fat tissues ratio was restored and glucose metabolism showed normal levels. Treating Fgfr3ach/+ mice with soluble FGFR3 during the growth period, prevented the development of these metabolic deregulations in adult animals and restored lean-over-fat tissues ratio as well as glucose metabolism in adult animals. Conclusion This study demonstrate that achondroplasia patients develop an atypical obesity with preferential abdominal obesity not associated with classical complications. These results suggest that achondroplasia induces an uncommon metabolism of energy, directly linked to the FGFR3 mutation. These data strongly suggest that this common complication of achondroplasia should be included in the clinical management of patients. In this context, sFGFR3 proved to be a promising treatment for achondroplasia by normalizing the biology at different levels, not only restoring bone growth but also preventing the atypical visceral obesity and some metabolic deregulations. PMID:29652901

Early postnatal soluble FGFR3 therapy prevents the atypical development of obesity in achondroplasia.

PubMed

Saint-Laurent, Celine; Garcia, Stephanie; Sarrazy, Vincent; Dumas, Karine; Authier, Florence; Sore, Sophie; Tran, Albert; Gual, Philippe; Gennero, Isabelle; Salles, Jean-Pierre; Gouze, Elvire

2018-01-01

Achondroplasia is a rare genetic disease is characterized by abnormal bone development and early obesity. While the bone aspect of the disease has been thoroughly studied, early obesity affecting approximately 50% of them during childhood has been somewhat neglected. It nevertheless represents a major health problem in these patients, and is associated to life-threatening complications including increasing risk of cardiovascular pathologies. We have thus decided to study obesity in patients and to use the mouse model to evaluate if soluble FGFR3 therapy, an innovative treatment approach for achondroplasia, could also impact the development of this significant complication. To achieve this, we have first fully characterized the metabolic deregulations in these patients by conducting a longitudinal retrospective study, in children with achondroplasia Anthropometric, densitometric measures as well as several blood parameters were recorded and compared between three age groups ranging from [0-3], [4-8] and [9-18] years old. Our results show unexpected results with the development of an atypical obesity with preferential fat deposition in the abdomen that is remarkably not associated with classical complications of obesity such as diabetes or hypercholosterolemia. Because it is not associated with diabetes, the atypical obesity has not been studied in the past even though it is recognized as a real problem in these patients. These results were validated in a murine model of achondroplasia (Fgfr3ach/+) where similar visceral adiposity was observed. Unexpected alterations in glucose metabolism were highlighted during high-fat diet. Glucose, insulin or lipid levels remained low, without the development of diabetes. Very interestingly, in achondroplasia mice treated with soluble FGFR3 during the growth period (from D3 to D22), the development of these metabolic deregulations was prevented in adult animals (between 4 and 14 weeks of age). The lean-over-fat tissues ratio was restored and glucose metabolism showed normal levels. Treating Fgfr3ach/+ mice with soluble FGFR3 during the growth period, prevented the development of these metabolic deregulations in adult animals and restored lean-over-fat tissues ratio as well as glucose metabolism in adult animals. This study demonstrate that achondroplasia patients develop an atypical obesity with preferential abdominal obesity not associated with classical complications. These results suggest that achondroplasia induces an uncommon metabolism of energy, directly linked to the FGFR3 mutation. These data strongly suggest that this common complication of achondroplasia should be included in the clinical management of patients. In this context, sFGFR3 proved to be a promising treatment for achondroplasia by normalizing the biology at different levels, not only restoring bone growth but also preventing the atypical visceral obesity and some metabolic deregulations.

Quantitative coronary angiography using image recovery techniques for background estimation in unsubtracted images

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

Wong, Jerry T.; Kamyar, Farzad; Molloi, Sabee

2007-10-15

Densitometry measurements have been performed previously using subtracted images. However, digital subtraction angiography (DSA) in coronary angiography is highly susceptible to misregistration artifacts due to the temporal separation of background and target images. Misregistration artifacts due to respiration and patient motion occur frequently, and organ motion is unavoidable. Quantitative densitometric techniques would be more clinically feasible if they could be implemented using unsubtracted images. The goal of this study is to evaluate image recovery techniques for densitometry measurements using unsubtracted images. A humanoid phantom and eight swine (25-35 kg) were used to evaluate the accuracy and precision of the followingmore » image recovery techniques: Local averaging (LA), morphological filtering (MF), linear interpolation (LI), and curvature-driven diffusion image inpainting (CDD). Images of iodinated vessel phantoms placed over the heart of the humanoid phantom or swine were acquired. In addition, coronary angiograms were obtained after power injections of a nonionic iodinated contrast solution in an in vivo swine study. Background signals were estimated and removed with LA, MF, LI, and CDD. Iodine masses in the vessel phantoms were quantified and compared to known amounts. Moreover, the total iodine in left anterior descending arteries was measured and compared with DSA measurements. In the humanoid phantom study, the average root mean square errors associated with quantifying iodine mass using LA and MF were approximately 6% and 9%, respectively. The corresponding average root mean square errors associated with quantifying iodine mass using LI and CDD were both approximately 3%. In the in vivo swine study, the root mean square errors associated with quantifying iodine in the vessel phantoms with LA and MF were approximately 5% and 12%, respectively. The corresponding average root mean square errors using LI and CDD were both 3%. The standard deviations in the differences between measured iodine mass in left anterior descending arteries using DSA and LA, MF, LI, or CDD were calculated. The standard deviations in the DSA-LA and DSA-MF differences (both {approx}21 mg) were approximately a factor of 3 greater than that of the DSA-LI and DSA-CDD differences (both {approx}7 mg). Local averaging and morphological filtering were considered inadequate for use in quantitative densitometry. Linear interpolation and curvature-driven diffusion image inpainting were found to be effective techniques for use with densitometry in quantifying iodine mass in vitro and in vivo. They can be used with unsubtracted images to estimate background anatomical signals and obtain accurate densitometry results. The high level of accuracy and precision in quantification associated with using LI and CDD suggests the potential of these techniques in applications where background mask images are difficult to obtain, such as lumen volume and blood flow quantification using coronary arteriography.« less

Vertebral Fracture Assessment by dual-energy X-ray absorptiometry: insurance coverage issues in the United States. A White Paper of the International Society for Clinical Densitometry.

PubMed

Laster, Andrew J; Lewiecki, E Michael

2007-01-01

Clinical trial data and fracture risk prediction models unequivocally demonstrate the utility of identifying prevalent vertebral fractures to predict future fractures of all types. Knowledge of prevalent vertebral fractures can alter patient management decisions and result in initiation of therapy to reduce fracture risk in some patients who would not otherwise be treated. Cost-benefit analysis demonstrates that identifying and treating patients with vertebral fractures, even those with a densitometric classification of osteopenia, is cost effective. Vertebral fractures can be readily identified in the office setting using standard radiography or Vertebral Fracture Assessment (VFA), a software addition to a central dual-energy X-ray absorptiometry (DXA) machine. In the United States, VFA was assigned a Current Procedural Terminology (CPT) code in January 2005. Nevertheless, coverage of VFA has not been uniformly embraced by Medicare carriers, companies that contract with the federal government to administer Medicare coverage and process claims for a region of the United States. Unlike DXA, for which uniform national coverage of qualified Medicare beneficiaries is mandated by the Balanced Budget Act of 1997, VFA coverage policies are determined by the local Medicare carriers. Third-party insurers are also variable in their coverage of VFA. This International Society for Clinical Densitometry (ISCD) White Paper documents the role of VFA in the evaluation and treatment of women with postmenopausal osteoporosis and compares it with standard spine radiography. Arguments used by some Medicare carriers and insurers to deny coverage of VFA in the United States are analyzed and critiqued. For health care providers within the United States, this White Paper may serve as a resource to respond to insurers who deny coverage of VFA. For health care providers regardless of their country, this article underscores the value of VFA as an alternative to spine radiography in the evaluation and management of postmenopausal women with suspected osteoporosis.

Doxycycline alters collagen composition following ventral hernia repair.

PubMed

Tharappel, Job C; Harris, Jennifer W; Totten, Crystal; Zwischenberger, Brittany A; Roth, John S

2017-04-01

Doxycycline, a nonspecific metalloproteinase (MMP) inhibitor, has been demonstrated to impact the strength of the polypropylene (PP) mesh-repaired hernia with an increase in the deposition of collagen type 1. The impact of doxycycline with porcine acellular dermal matrices (PADM) is unknown; therefore, we evaluated the impact of doxycycline administration upon hernia repair with PP and PADM mesh. Sprague-Dawley rats weighing ~400 g underwent laparotomy with creation of a midline ventral hernia. After a 27-day recovery, animals were randomly assigned to four groups of eight and underwent intraperitoneal underlay hernia repair with either PP or PADM. Groups were assigned to daily normal saline (S) or daily doxycycline in normal saline 10 mg/kg (D) via oral gavage for 8 weeks beginning 24 h preoperatively. Animals were euthanized at 8 weeks and underwent tensiometric testing of the abdominal wall and western blot analyses for collagen subtypes and MMPs. Thirty-two animals underwent successful hernia creation and repair with either PADM or PP. At 8 weeks, 15 of 16 PP-implanted animals survived with only 12 of 16 PADM-implanted animals surviving. There were no differences in the mesh to fascial interface tensiometric strength between groups. Densitometric counts in the PADM-D group demonstrated increased collagen type 1 compared to PP-S (PADM-D [1286.5], PADM-S [906.9], PP-S [700.4], p = 0.037) and decreased collagen type 3 compared to PP-S (PADM-D [7446.9], PADM-S [8507.6], PP-S [11,297.1], p = 0.01). MMP-9 levels were increased in PADM-D (PP-S vs. PADM-D, p = 0.04), while MMP-2 levels were similar between PADM-D and PADM-S, respectively. Collagen type 1 deposition at the mesh to fascial interface is enhanced following administration of doxycycline in ventral hernia repairs with porcine acellular dermal matrices. Doxycycline administration may have implications for enhancing hernia repair outcomes using biologic mesh.

Effects of Two Training Modalities on Body Fat and Insulin Resistance in Postmenopausal Women.

PubMed

Henríquez, Sandra; Monsalves-Alvarez, Matías; Jimenez, Teresa; Barrera, Gladys; Hirsch, Sandra; de la Maza, María Pia; Leiva, Laura; Rodriguez, Juan Manuel; Silva, Claudio; Bunout, Daniel

2017-11-01

Henríquez, S, Monsalves-Alvarez, M, Jimenez, T, Barrera, G, Hirsch, S, de la Maza, MP, Leiva, L, Rodriguez, JM, Silva, C, and Bunout, D. Effects of two training modalities on body fat and insulin resistance in postmenopausal women. J Strength Cond Res 31(11): 2955-2964, 2017-Our objective was to compare the effects of a low-load circuit resistance training protocol and usual aerobic training in postmenopausal women. Postmenopausal women with at least 1 feature of the metabolic syndrome were randomly allocated to a low-load circuit resistance training protocol or traditional aerobic training in a braked cycle ergometer. The intervention consisted in supervised sessions lasting 40 minutes, 3 times per week, during 6 months. At baseline and at the end of the intervention, fasting serum lipid levels, serum interleukin 6, C-reactive protein, 8 isoprostanes, and insulin resistance (assessed through QUICKI and HOMA-IR) were measured. Body fat was measured by double-beam X-ray absorptiometry and by computed tomography densitometric quantification at lumbar 3 vertebral level. Twenty-one women aged 58 (54-59) years were allocated to aerobic training and 21 women aged 55 (52-61) years were allocated to the low-load circuit resistance training protocol. Eighteen and 16 women in each group completed the 6 months training period. Women in both groups experienced significant reductions in blood pressure, total body, subcutaneous, and intraabdominal body fat. Reductions in total cholesterol and triacylglycerol levels were also observed. No changes in insulin resistance indexes, 8 isoprostanes, C-reactive protein, or interleukin 6 were observed in either group. No significant differences between treatment groups were observed in any of the measured parameters. We conclude that low-load circuit resistance training and aerobic training resulted in the same reductions in body fat and serum lipid levels.

Serum protein electrophoresis: potential test for use in geriatric companion animal health programmes.

PubMed

Batamuzi, E K; Kristensen, E; Jensen, A L

1996-10-01

The electrophoretic patterns of 117 serum samples from 37 clinically healthy old dogs (age: 10.0 +/- 2.5 years, mean +/- SD) 47 old dogs (age: 9.7 +/- 2.0 years) with different diseases, 18 clinically healthy young dogs (age: 2.9 +/- 1.6 years) and 15 young dogs (age: 2.0 +/- 1.0 years) with different diseases were evaluated using agarose as a supporting matrix. Three major electrophoretic patterns were identified after densitometric scanning. The first pattern was a predominant pattern for the young healthy dogs (found in 67% of dogs in that group) and was considered normal for healthy dogs. The second pattern had relatively higher proportions of beta and gamma globulins (polyclonal immunoglobulinaemia) and was a predominant pattern among the old healthy dogs (found in 51% of dogs in that group). The findings regarding the second pattern appear to suggest that they had been exposed to antigens during their lives and that they had a well developed and responsive humoral immune response. The third pattern was characterized by high levels of alpha globulins, it was the most common pattern in the old diseased dogs (found in 70% of the dogs in that group). The third pattern was considered to indicate an inflammatory response among old dogs when compared to young dogs (P < 0.01). These patterns appear to suggest that because of aging and/or accumulated injury geriatric dogs will have high levels of polyclonal gamma-globulinaemia or acute phase reactants. The observed patterns, especially the one showing high levels of alpha-globulins, could be a valuable parameter in the search for individuals requiring special attention and thus for the establishment of health programmes for geriatric dogs. With further effort, serum protein electrophoresis might prove to be a useful test for the evaluation of geriatric companion animals for diagnostic, prognostic and therapeutic reasons.

IGF-1 and Survival in ESRD

PubMed Central

Jia, Ting; Gama Axelsson, Thiane; Heimbürger, Olof; Bárány, Peter; Stenvinkel, Peter; Qureshi, Abdul Rashid

2014-01-01

Summary Background and objectives IGF-1 deficiency links to malnutrition in CKD patients; however, it is not clear to what extent it associates with survival among these patients. Design, setting, participants, & measurements Serum IGF-1 and other biochemical, clinical (subjective global assessment), and densitometric (dual energy x-ray absorptiometry) markers of nutritional status and mineral and bone metabolism were measured in a cohort of 365 Swedish clinically stable CKD stage 5 patients (median age of 53 years) initiating dialysis between 1994 and 2009; in 207 patients, measurements were also taken after 1 year of dialysis. Deaths were registered during a median follow-up of 5 years. Associations of mortality with baseline IGF-1 and changes of IGF-1 after 1 year of dialysis were evaluated by Cox models. Results At baseline, IGF-1 concentrations associated negatively with age, diabetes mellitus, cardiovascular disease, poor nutritional status, IL-6, and osteoprotegerin and positively with body fat mass, bone mineral density, serum phosphate, calcium, and fibroblast growth factor-23. At 1 year, IGF-1 had increased by 33%. In multivariate regression, low age, diabetes mellitus, and high serum phosphate and calcium associated with IGF-1 at baseline, and in a mixed model, these factors, together with high fat body mass, associated with changes of IGF-1 during the first 1 year of dialysis. Adjusting for calendar year of inclusion, age, sex, diabetes mellitus, cardiovascular disease, IL-6, and poor nutritional status, a 1 SD higher level of IGF-1 at baseline associated with lower mortality risk (hazard ratio, 0.57; 95% confidence interval, 0.32 to 0.98). Persistently low or decreasing IGF-1 levels during the first 1 year on dialysis predicted worse survival (adjusted hazard ratio, 2.19; 95% confidence interval, 1.06 to 4.50). Conclusion In incident dialysis patients, low serum IGF-1 associates with body composition and markers of mineral and bone metabolism, and it predicts increased mortality risk. PMID:24178975

TLC-Densitometric and RP-HPLC Methods for Simultaneous Determination of Dexamethasone and Chlorpheniramine Maleate in the Presence of Methylparaben and Propylparaben.

PubMed

Farid, Nehal F; Naguib, Ibrahim A; Moatamed, Radwa S; El Ghobashy, Mohamed R

2017-01-01

Validated simple, sensitive, and highly selective methods are applied for the quantitative determination of dexamethasone and chlorpheniramine maleate in the presence of their reported preservatives (methylparaben and propylparaben), whether in pure forms or in pharmaceutical formulation. TLC is the first method, in which dexamethasone, chlorpheniramine maleate, methylparaben, and propylparaben are separated on silica gel TLC F254 plates using hexane-acetone-ammonia (5.5 + 4.5 + 0.5, v/v/v) as the developing phase. Separated bands are scanned at 254 nm over a concentration range of 0.1-1.7 and 0.4-2.8 μg/band, with mean ± SD recoveries of 99.12 ± 0.964 and 100.14 ± 0.962%, for dexamethasone and chlorpheniramine maleate, respectively. Reversed-phase HPLC is the second method, in which a mixture of dexamethasone and chlorpheniramine maleate, methylparaben, and propylparaben is separated on a reversed-phase silica C18 (5 μm particle size, 250 mm, 4.6 mm id) column using 0.1 M ammonium acetate buffer-acetonitrile (60 + 40, v/v, pH 3) as the mobile phase. The drugs were detected at 220 nm over a concentration range of 5-50 μg/mL, 2-90 μg/mL, 4-100 μg/mL, and 7-50 μg/mL, with mean ± SD recoveries of 100.85 ± 0.905, 99.67 ± 1.281, 100.20 ± 0.906, and 99.81 ± 0.954%, for dexamethasone, chlorpheniramine maleate, methylparaben paraben, and propylparaben, respectively. The advantages of the suggested methods over previously reported methods are the ability to detect lower concentrations of the main drugs and to show better resolution of interfering preservatives; hence, these methods could be more reliable for routine QC analyses.

Inter-species comparative antioxidant assay and HPTLC analysis of sakuranetin in the chloroform and ethanol extracts of aerial parts of Rhus retinorrhoea and Rhus tripartita.

PubMed

Alam, Perwez; Parvez, Mohammad Khalid; Arbab, Ahmed Hassan; Siddiqui, Nasir Ali; Al-Dosary, Mohammed Salem; Al-Rehaily, Adnan Jathlan; Ahmed, Sarfaraz; Kalam, Mohd Abul; Ahmad, Mohammad Shamim

2017-12-01

Extensive research on Rhus (Anacardiaceae) shows their antioxidant potential, which warrants further evaluation of its other species. To perform a comparative antioxidant assay on extracts of R. retinorrhoea and R. tripartita, including sakuranetin quantification by a validated HPTLC method. In vitro antioxidant assay was performed on chloroform and ethanol extracts of R. retinorrhoea Steud. ex Oliv. (RRCE and RREE) and R. tripartita (Ucria) Grande (RTCE and RTEE) by DPPH radical scavenging (at 31.25, 62.5, 125, 250 and 500 μg/mL concentrations) and β-carotene-linoleic acid bleaching methods at 500 μg/mL concentration. Densitometric HPTLC method was developed and validated using toluene: ethyl acetate: methanol (8:2:0.2; v/v/v) as mobile phase, executed on glass-backed silica gel F 254 plate and scanned at 292 nm. Antioxidant activity of Rhus extracts tested by the two methods (DPPH/BCB) was found in order of RTEE > RREE > RTCE > RRCE with IC 50 118.67/256.26, 315.75/82.35, 827.92/380.0 and 443.69/292.75, respectively. Scanning of the HPTLC plate provided an intense peak of sakuranetin at R f = 0.59. The estimated sakuranetin content in the dry weight of the extracts was highest in RREE (27.95 μg/mg) followed by RRCE (25.22 μg/mg), RTEE (0.487 μg/mg) and RTCE (0.0 μg/mg). Presence of sakuranetin in RREE, RRCE and RTEE supported the highest antioxidant property of the two Rhus species. Nonetheless, low sakuratenin in R. tripartita indicated the presence of other bioactive constituents responsible for synergistic antioxidant activity. The developed HPTLC method therefore guarantees its application in quality control of commercialized herbal drugs and formulations containing sakuranetin.

Environmental impacts on the developing CNS: CD15, NCAM-L1, and GFAP expression in rat neonates exposed to hypergravity

NASA Technical Reports Server (NTRS)

Sulkowski, G. M.; Li, G-H; Sajdel-Sulkowska, E. M.

2004-01-01

We have previously reported that the developing rat cerebellum is affected by hypergravity exposure. The effect is observed during a period of both granule and glial cell proliferation and neuronal migration in the cerebellum and coincides with changes in thyroid hormone levels. The present study begins to address the molecular mechanisms involved in the cerebellar response to hypergravity. Specifically, the study focuses on the expression of cerebellar proteins that are known to be directly involved in cell-cell interactions [protein expressing 3-fucosyl-N-acetyl-lactosamine antigen (CD15), neuronal cell adhesion molecule (NCAM-L1)] and those that affect cell-cell interactions indirectly [glial fibrillary acidic protein (GFAP)] in rat neonates exposed to centrifuge-produced hypergravity. Cerebellar mass and protein expression in rat neonates exposed to hypergravity (1.5 G) from gestational day (G) 11 to postnatal day (P) 30 were compared at one of six time points between P6 and P30 against rat neonates developing under normal gravity. Proteins were analyzed by quantitative western blots of cerebellar homogenates prepared from male or female neonates. Cerebellar size was most clearly reduced in male neonates on P6 and in female neonates on P9, with a significant gender difference; differences in cerebellar mass remained significant even when change in total body mass was factored in. Densitometric analysis of western blots revealed both quantitative and temporal changes in the expression of selected cerebellar proteins that coincided with changes in cerebellar mass and were gender-specific. In fact, our data indicated certain significant differences even between male and female control animals. A maximal decrease in expression of CD15 was observed in HG females on P9, coinciding with maximal change in their cerebellar mass. A shift in the time-course of NCAM-L1 expression resulted in a significant increase in NCAM-L1 in HG males on P18, an isolated time at which cerebellar mass does not significantly differ between HG and SC neonates. A maximal decrease in expression of GFAP was observed in HG males on P6, coinciding with maximal change in their cerebellar mass. Altered expression of cerebellar proteins is likely to affect a number of developmental processes and contribute to the structural and functional alterations seen in the CNS developing under altered gravity. Our data suggest that both cerebellar development and its response to gravitational manipulations differ in males and females. c2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

Environmental impacts on the developing CNS: CD15, NCAM-L1, and GFAP expression in rat neonates exposed to hypergravity

NASA Astrophysics Data System (ADS)

Sulkowski, G. M.; Li, G.-H.; Sajdel-Sulkowska, E. M.

2004-01-01

We have previously reported that the developing rat cerebellum is affected by hypergravity exposure. The effect is observed during a period of both granule and glial cell proliferation and neuronal migration in the cerebellum and coincides with changes in thyroid hormone levels. The present study begins to address the molecular mechanisms involved in the cerebellar response to hypergravity. Specifically, the study focuses on the expression of cerebellar proteins that are known to be directly involved in cell-cell interactions [protein expressing 3-fucosyl- N-acetyl-lactosamine antigen (CD15), neuronal cell adhesion molecule (NCAM-L1)] and those that affect cell-cell interactions indirectly [glial fibrillary acidic protein (GFAP)] in rat neonates exposed to centrifuge-produced hypergravity. Cerebellar mass and protein expression in rat neonates exposed to hypergravity (1.5 G) from gestational day (G) 11 to postnatal day (P) 30 were compared at one of six time points between P6 and P30 against rat neonates developing under normal gravity. Proteins were analyzed by quantitative western blots of cerebellar homogenates prepared from male or female neonates. Cerebellar size was most clearly reduced in male neonates on P6 and in female neonates on P9, with a significant gender difference; differences in cerebellar mass remained significant even when change in total body mass was factored in. Densitometric analysis of western blots revealed both quantitative and temporal changes in the expression of selected cerebellar proteins that coincided with changes in cerebellar mass and were gender-specific. In fact, our data indicated certain significant differences even between male and female control animals. A maximal decrease in expression of CD15 was observed in HG females on P9, coinciding with maximal change in their cerebellar mass. A shift in the time-course of NCAM-L1 expression resulted in a significant increase in NCAM-L1 in HG males on P18, an isolated time at which cerebellar mass does not significantly differ between HG and SC neonates. A maximal decrease in expression of GFAP was observed in HG males on P6, coinciding with maximal change in their cerebellar mass. Altered expression of cerebellar proteins is likely to affect a number of developmental processes and contribute to the structural and functional alterations seen in the CNS developing under altered gravity. Our data suggest that both cerebellar development and its response to gravitational manipulations differ in males and females.

TRAIL-induced programmed necrosis as a novel approach to eliminate tumor cells

PubMed Central

2014-01-01

Background The cytokine TRAIL represents one of the most promising candidates for the apoptotic elimination of tumor cells, either alone or in combination therapies. However, its efficacy is often limited by intrinsic or acquired resistance of tumor cells to apoptosis. Programmed necrosis is an alternative, molecularly distinct mode of programmed cell death that is elicited by TRAIL under conditions when the classical apoptosis machinery fails or is actively inhibited. The potential of TRAIL-induced programmed necrosis in tumor therapy is, however, almost completely uncharacterized. We therefore investigated its impact on a panel of tumor cell lines of wide-ranging origin. Methods Cell death/viability was measured by flow cytometry/determination of intracellular ATP levels/crystal violet staining. Cell surface expression of TRAIL receptors was detected by flow cytometry, expression of proteins by Western blot. Ceramide levels were quantified by high-performance thin layer chromatography and densitometric analysis, clonogenic survival of cells was determined by crystal violet staining or by soft agarose cloning. Results TRAIL-induced programmed necrosis killed eight out of 14 tumor cell lines. Clonogenic survival was reduced in all sensitive and even one resistant cell lines tested. TRAIL synergized with chemotherapeutics in killing tumor cell lines by programmed necrosis, enhancing their effect in eight out of 10 tested tumor cell lines and in 41 out of 80 chemotherapeutic/TRAIL combinations. Susceptibility/resistance of the investigated tumor cell lines to programmed necrosis seems to primarily depend on expression of the pro-necrotic kinase RIPK3 rather than the related kinase RIPK1 or cell surface expression of TRAIL receptors. Furthermore, interference with production of the lipid ceramide protected all tested tumor cell lines. Conclusions Our study provides evidence that TRAIL-induced programmed necrosis represents a feasible approach for the elimination of tumor cells, and that this treatment may represent a promising new option for the future development of combination therapies. Our data also suggest that RIPK3 expression may serve as a potential predictive marker for the sensitivity of tumor cells to programmed necrosis and extend the previously established role of ceramide as a key mediator of death receptor-induced programmed necrosis (and thus as a potential target for future therapies) also to the tumor cell lines examined here. PMID:24507727

Lung cancer pathogenesis associated with wood smoke exposure.

PubMed

Delgado, Javier; Martinez, Luis M; Sánchez, Therasa T; Ramirez, Alejandra; Iturria, Cecilia; González-Avila, Georgina

2005-07-01

Tobacco is considered the most important cause of lung cancer, but other factors could also be involved in its pathogenesis. The aim of the present work was to establish an association between wood smoke exposure and lung cancer pathogenesis, and to analyze the effects of wood smoke on p53 and murine double minute 2 (MDM2) protein expression. Blood samples were obtained from 62 lung cancer patients, 9 COPD patients, and 9 control subjects. Of the 62 lung cancer patients, 23 were tobacco smokers (lung cancer associated with tobacco [LCT] group), 24 were exposed to wood smoke (lung cancer associated with wood smoke [LCW] group), and 15 could not be included in these groups. Western blot assays were performed to identify the presence of p53, phospho-p53, and murine double minute 2 (MDM2) isoforms in plasma samples. Densitometric analysis was used to determine the intensity of p53, phospho-p53, and MDM2 bands. Approximately 38.7% of the lung cancer patients examined had an association with wood smoke exposure, most of them women living in rural areas. Adenocarcinoma was present in 46.7% of these patients. The p53 and phospho-p53 proteins were significantly increased in LCW samples (56,536.8 +/- 4,629 densitometry units [DU] and 58,244.8 +/- 7,492 DU, respectively [+/- SD]), in comparison with the other groups. The 57-kD MDM2 isoform plasma concentration was very high in LCW and LCT samples (75,696.4 +/- 11,979 DU and 78,551.7 +/- 11,548 DU, respectively). MDM2-p53 complexes were present in a high concentration in control and COPD subjects. This allows p53 degradation and explains the low concentrations of p53 found in these groups. MDM2-phospho-p53 complexes were observed in COPD but not in the other samples. This correlates with the low concentration of p53 observed in the COPD group (13,657 +/- 2,012 DU), and could explain the different clinic evolution of this smoker population in comparison with the LCT subjects. This study suggests that there is a possible association of lung cancer with wood smoke exposure. Likewise, our findings demonstrate that wood smoke could produce similar effects on p53, phospho-p53, and MDM2 protein expression as tobacco.

Simultaneous Quantification of Forskolin and Iso-Forskolin in Coleus forskohlii (Wild.) Briq. and Identification of Elite Chemotype, Collected from Eastern Ghats (India)

PubMed Central

Shukla, Pushpendra Kumar; Misra, Ankita; Kumar, Manish; Jaichand; Singh, Kuldeep; Akhtar, Juber; Srivastava, Sharad; Agrawal, Pawan K; Singh Rawat, Ajay K

2017-01-01

Background: Coleus forskohlii is a well-known industrially important medicinal plant, for its high forskolin content. Objective: A simple, selective, and sensitive high-performance thin layer chromatography (HPTLC) method was developed and validated for simultaneous quantification of forskolin and iso-forskolin in C. forskohlii germplasm collected from the Eastern Ghats, India. Materials and Methods: Chromatographic separation of the targeted marker(s) was obtained on precoated silica plates using toluene: ethyl acetate: methanol (90:30:0.5, v/v/v) as the mobile phase. Results: Densitometric quantification of forskolin and iso-forskolin was carried out at 545 nm. Forskolin and iso-forskolin were identified by comparing the ultraviolet spectra of standard and sample track at Rf of 0.64 ± 0.02 and 0.36 ± 0.01, after derivatization with anisaldehyde sulfuric acid reagent. The linearity of both the analytes was obtained in the range of 300–1200 ng/spot with the regression coefficient (R2) of 0.991 and 0.986. Recovery of analyte (s) at three levels, namely, 100, 150, and 200 ng/spot was found to be 100.46% ± 0.29%, 99.64% ± 0.33%, 100.02% ± 0.76% and 99.76% ± 0.62%, 99.56% ± 0.35%, 100.02% ± 0.22%, respectively, for forskolin and iso-forskolin. The content of forskolin and iso-forskolin varies from 0.046% to 0.187% and 0.002% to 0.077%, respectively (dry weight basis), the maximum content of both the markers was found in NBC-31, from Thakurwada, Maharashtra. Conclusion: The developed HPTLC method was linear, accurate, and reliable as per the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The study aids in the identification of elite chemotype for commercial prospection of industrially viable medicinal crop. SUMMARY 12 Samples are collected from different locations of the eastern ghat regionsQuantification of two major marker forskolin and iso forskolinThe maximum content of both the markers was found in NBC -31, from Thakurwada, MaharashtraIdentification of elite chemotype of collected samples may be useful for commercial prospection in industries. PMID:29491648

Melatonin accelerates maturation inducing hormone (MIH): induced oocyte maturation in carps.

PubMed

Chattoraj, Asamanja; Bhattacharyya, Sharmistha; Basu, Dipanjan; Bhattacharya, Shelley; Bhattacharya, Samir; Maitra, Saumen Kumar

2005-02-01

The present communication is an attempt to demonstrate the influence of melatonin on the action of maturation inducing hormone (MIH) on the maturation of oocytes in carps. The oocytes from gravid female major carp Labeo rohita were isolated and incubated separately in Medium 199 containing (a) only MIH (1 microg/ml), (b) only melatonin (at concentrations of 50, 100 or 500 pg/ml), and (c) both melatonin and MIH, but at different time intervals. In the latter group, melatonin was added to the incubating medium either (i) 4 h before addition of MIH, (ii) 2 h before addition of MIH, (iii) co-administered with MIH (0 h interval) or (iv) 2 h after addition of MIH. In each case, oocytes were further incubated for 4, 8, 12 or 16 h post- administration of MIH, and the effects of treatment on oocyte maturation were evaluated by considering the rate (%) of germinal vesicle breakdown (GVBD). Incubation of oocytes in a medium containing only melatonin did not result in GVBD of any oocyte. Nearly all the oocytes underwent GVBD when incubated with MIH for 16 h. Administration of melatonin along with MIH (at 0 h interval) or 2 h after addition of MIH did not result in any significant change in the rate of GVBD compared to that in a medium containing only MIH. However, it was quite interesting to observe that incubation of oocytes with melatonin especially 4 h prior to addition of MIH in the medium, led to an accelerated rate of GVBD in the oocytes. Experiments with the oocytes of another major carp Cyprinus carpio following an identical schedule depicted similar results except a difference in the optimum melatonin dose. In L. rohita, 50 pg/ml melatonin had maximum acceleratory effect on MIH-induced GVBD of oocytes, while it was 100 pg/ml in C. carpio. Further study revealed that pre-incubation with melatonin accelerates the action of MIH on the formation of a complex of two proteins (MPF), a regulatory component called cyclin B and the catalytic component protein kinase known as cyclin-dependent kinase, Cdk1. Densitometric analysis of the immunoblot data collected from the melatonin pre-treated MIH incubated oocytes showed that cyclin B level continued to increase even after 4 h of incubation, and reached the peak after 12 h. Moreover, determination of H1 kinase activity as an indicator of MPF activity in oocytes revealed that melatonin pre-incubation considerably increased MIH stimulation of histone H1 phosphorylation as compared to MIH alone. Thus, the present study demonstrates for the first time that prior incubation with melatonin accelerates the action of MIH on carp oocyte maturation.

Controlling systematic perioperative anaerobic contamination during sinus-lift procedures by using metronidazole: an innovative approach.

PubMed

Choukroun, Joseph; Simonpieri, Alain; Del Corso, Marco; Mazor, Ziv; Sammartino, Gilberto; Dohan Ehrenfest, David M

2008-09-01

Analysis of tomodensitometric controls following sinus grafts clearly demonstrates a quite systematic lack of homogeneity. Sinus contamination by anaerobic bacteria seems almost unavoidable during bone graft surgery, and this problem may jeopardize the healing process. The aim of this study was to characterize in a systematic way the nonhomogeneities observed at 1, 2, or 3 months postsurgery within allogenous sinus grafts, and to assess the possible influence of a 0.5% sterile solution of metronidazole incorporated in the sinus bone graft. This clinical study was conducted on 72 patients treated with single or bilateral sinus-lifts: 94 sinus elevations performed with freeze-dried bone allograft (Phoenix, TBF, Mions, France), with (test group) or without (control group) metronidazole. In the test group, each bone graft was hydrated with 2 mL of a 0.5% metronidazole solution, i.e., only 10 mg of metronidazole. All the patients went through a first presurgical computerized tomography (CT)-scan followed by a second scan performed at 1, 2, or 3 months postsurgery (which was used as the preimplant reference scan). For 11 patients, 2 postsurgical CT-scans were performed respectively at 10 days and 2 months. Using an arbitrary gray scale (Arbitrary Densitometric Unit) which functions according to the Hounsfield unit principle, the degree of radiographic homogeneity of the grafts was established. Density scattering provides some information on the homogeneity or nonhomogeneity of the bone graft. The 12 grafts performed without metronidazole show significant nonhomogeneities at 1, 2, or 3 months. Moreover, when a CT-scan is performed during the first postoperative days (at 10 days), the presence of air bubbles in the graft is confirmed. The tomodensitometric aspects of all grafts treated with metronidazole in this series are absolutely identical: they show a high degree of homogeneity. Sixty-three cases (76.8%) are homogeneous, and 19 cases (23.2%) are significantly homogeneous. The time at which the control scan is performed (10 days, 1, 2, or 3 months) does not seem to influence significantly the degree of homogeneity assessed. In the control group, some inflammatory events associated with facial oedema were observed in 25% of the cases. In the test group, no such event was recorded for the 82 sinus-lifts treated with metronidazole. A possible correlation may exist between the occurrence of non homogeneities within the bone grafts and the anaerobic bacterial contamination. The local use of a very small quantity of metronidazole (equivalent to only 1/20 of a common 200 mg oral tablet) could provide more security when performing sinus-lift procedures and an improved quality of the graft. This protocol should not be considered as an antibiotherapy, but only as way to limit the initial contamination of bone graft.

Advantages of a validated UPLC-MS/MS standard addition method for the quantification of A-type dimeric and trimeric proanthocyanidins in cranberry extracts in comparison with well-known quantification methods.

PubMed

van Dooren, Ines; Foubert, Kenn; Theunis, Mart; Naessens, Tania; Pieters, Luc; Apers, Sandra

2018-01-30

The berries of Vaccinium macrocarpon, cranberry, are widely used for the prevention of urinary tract infections. This species contains A-type proanthocyanidins (PACs), which intervene in the initial phase of the development of urinary tract infections by preventing the adherence of Escherichia coli by their P-type fimbriae to uroepithelial cells. Unfortunately, the existing clinical studies used different cranberry preparations, which were poorly standardized. Because of this, the results were hard to compare, which led sometimes to conflicting results. Currently, PACs are quantified using the rather non-specific spectrophotometric 4-dimethylaminocinnamaldehyde (DMAC) method. In addition, a normal phase HPTLC-densitometric method, a HPLC-UV method and three LC-MS/MS methods for quantification of procyanidin A2 were recently published. All these methods contain some shortcomings and errors. Hence, the development and validation of a fast and sensitive standard addition LC-MS/MS method for the simultaneous quantification of A-type dimers and trimers in a cranberry dry extract was carried out. A linear calibration model could be adopted for dimers and, after logaritmic transformation, for trimers. The maximal interday and interconcentration precision was found to be 4.86% and 4.28% for procyanidin A2, and 5.61% and 7.65% for trimeric PACs, which are all acceptable values for an analytical method using LC-MS/MS. In addition, twelve different cranberry extracts were analyzed by means of the newly validated method and other widely used methods. There appeared to be an enormous variation in dimeric and trimeric PAC content. Comparison of these results with LC-MS/MS analysis without standard addition showed the presence of matrix effects for some of the extracts and proved the necessity of standard addition. A comparison of the well-known and widely used DMAC method, the butanol-HCl assay and this newly developed LC-MS/MS method clearly indicated the need for a reliable method able to quantify A-type PACs, which are considered to be the pharmacologically active constituents of cranberry, since neither the DMAC or butanol-HCl assays are capable of distinguishing between A and B-type PACs and therefore cannot detect adulterations with, for example, extracts with a high B-type PAC content. Hence, the combination of the DMAC method or butanol-HCl assay with this more specific LC-MS/MS assay could overcome these shortcomings. Copyright © 2017 Elsevier B.V. All rights reserved.

Trees tell of past climates: but are they speaking less clearly today?

PubMed Central

Briffa, K. R.

1998-01-01

The annual growth of trees, as represented by a variety of ring-width, densitometric, or chemical parameters, represents a combined record of different environmental forcings, one of which is climate. Along with climate, relatively large-scale positive growth influences such as hypothesized 'fertilization' due to increased levels of atmospheric carbon dioxide or various nitrogenous compounds, or possibly deleterious effects of 'acid rain' or increased ultra-violet radiation, might all be expected to exert some influence on recent tree growth rates. Inferring the details of past climate variability from tree-ring data remains a largely empirical exercise, but one that goes hand-in-hand with the development of techniques that seek to identify and isolate the confounding influence of local and larger-scale non-climatic factors. By judicious sampling, and the use of rigorous statistical procedures, dendroclimatology has provided unique insight into the nature of past climate variability, but most significantly at interannual, decadal, and centennial timescales. Here, examples are shown that illustrate the reconstruction of annually resolved patterns of past summer temperature around the Northern Hemisphere, as well as some more localized reconstructions, but ones which span 1000 years or more. These data provide the means of exploring the possible role of different climate forcings; for example, they provide evidence of the large-scale effects of explosive volcanic eruptions on regional and hemispheric temperatures during the last 400 years. However, a dramatic change in the sensitivity of hemispheric tree-growth to temperature forcing has become apparent during recent decades, and there is additional evidence of major tree-growth (and hence, probably, ecosystem biomass) increases in the northern boreal forests, most clearly over the last century. These possibly anthropogenically related changes in the ecology of tree growth have important implications for modelling future atmospheric CO2 concentrations. Also, where dendroclimatology is concerned to reconstruct longer (increasingly above centennial) temperature histories, such alterations of 'normal' (pre-industrial) tree-growth rates and climate-growth relationships must be accounted for in our attempts to translate the evidence of past tree growth changes.

A Clinico-analytical Study on Seed of Wrightia antidysenterica Linn. as a Therapeutic Emetic Agent (Vamaka Yoga) in the Management of Psoriasis.

PubMed

Bhattacharyya, Nirupam; Pujar, Muralidhar P; Chaturvedi, Ashutosh; Kumar, M Ashvini; Lohith, B A; Kumar, K N Sunil

2016-03-01

Wrightia antidysenterica Linn. (WA) is male variety Kutaja stated to be potent therapeutic emetic agent in skin disorders. Expulsion of doshas through oral route is termed as Vamana Karma (VK) (therapeutic emesis). However, so far, its utility for Vamana is not explored in detail, therefore there is a need to revalidate the utility of WA for Vamana. Hence, the above study was conducted to ascertain the efficacy as a therapeutic emetic agent (vamaka yoga) in the management of psoriasis along with quality control and standardization of this herb. The drug was standardized as per analytical procedures in Pharmacopeias. Thirty patients of psoriasis fulfilling inclusion criteria were taken for the study and Vamana with WA was conducted. Criteria were prepared to assess the signs and Symptoms of psoriasis. VK was assessed using the classical Lakshanas (features) such as Anthiki shudhi (Ending symptoms of emesis), Vaigiki shudhi (features of vomiting bouts), Maniki shudhi (Quantitative and qualitative purification), complications. VK with WA showed significant relief in parameters of psoriasis such as scaling, itching, candle grease sign (P < 0.001), and psoriasis area and severity index score (P = 0.001). In VK with WA, mean number of Vegas (vomiting bouts) was 6.91. 66% patients showing quantitative purification between 301 and 600 ml. 73.33% showed all Symptoms of purification. 73.33% patients showed Kaphanta vamana (Moderate expulsion of desire humor). In the level of biopurification, 66.66% patients showed moderated purification. No complication was noted with moderate drug palatability. Pharmacopeial analytical study showed its standardized values for testing the drug used for the study. It is proved as potent therapeutic emetic agent with no complication showed its clinical benefits over skin disorder like psoriasis. Seeds of Wrightia antidysenterica (WA) Linn. free from any foreign matter were selected for the study. Loss on drying revealed 6.535% moisture content; total ash indicating of total inorganic content was found to be 5.12%; acid insoluble ash is the acid insoluble part of total ash, mainly silica, WA showed 0.393% acid insoluble ash; ethanol and water soluble extractive is indicative of percentage active constituents were found to be 25.66 and 20.854%, respectively. High-performance thin layer chromatography fingerprinting profiles of WA under 254 nm showed the presence of 7 spots (all in green) at Rf values ranging from 0.21 to 0.88. Under 366 nm there were 4 prominent spots (all in fluorescent) at Rf 0.49 to 0.82 and, when scanned under white light 620 nm following derivatization with vanillin sulfuric acid 6 spots (in different colors) were evident at Rf 0.28 to 0.58. Among these spot with Rf of 0.58 was common when visualized under all the three methods. Rf values by densitometric scan of WA showed 12 peaks at 254 nm and 5 peaks at 366 nm. However, in clinical trial, it was found to be a potent emetic agent without any complication. Abbreviations Used: WA: Wrightia antidysenterica; Linn.; VK: Vamana karma; BT: Before treatment; FP: Freidman's P value; CHS: Chi-square value; NR: Negative ranks; PR: Positive ranks; N: Sample number, AS: Austipz sign; CG: Candle grease test; SSL: Samyak Snigdha Lakshana.

Improved efficacy of intramuscular weekly administration of clodronate 200 mg (100 mg twice weekly) compared with 100 mg (once weekly) for increasing bone mineral density in postmenopausal osteoporosis.

PubMed

Frediani, Bruno; Bertoldi, Ilaria; Pierguidi, Serena; Nicosia, Antonella; Picerno, Valentina; Filippou, Georgios; Cantarini, Luca; Galeazzi, Mauro

2013-03-01

Clodronate is a bisphosphonate used for the treatment of postmenopausal osteoporosis and all conditions characterized by excess bone resorption. We have previously reported that intramuscular (IM) therapy with clodronate at a dose of 100 mg/week displays significant effects on bone mineral density (BMD) although a plateau effect is observed after 1 year of treatment. Previous reports indicate that the densitometric effects of bisphosphonates directly correlate with the drug dosage and suggest that using IM clodronate at doses higher than 100 mg/week may result in improved efficacy. However, to the best of our knowledge, this has never been proved. The primary endpoint of the study was the effect on BMD of IM clodronate 100 mg once weekly or 100 mg twice weekly in patients with postmenopausal osteoporosis. The incidence of non-traumatic vertebral fractures and adverse events was also reported. The present study was a randomized, open-label, parallel-group trial conducted between January 2007 and December 2009 in the Osteoporosis and Osteoarticular Instrumental Diagnosis Centre (University of Siena, Siena, Italy). The study involved 60 women, aged 57-78 years, with a history of postmenopausal osteoporosis for more than 5 years. Patients were randomized to receive IM clodronate 100 mg once weekly (Group A, 30 patients) or 100 mg twice weekly (Group B, 30 patients), for 2 years. Significant increases compared with baseline in BMD were observed for both groups at 1 and 2 years, with significantly higher increases for Group B compared with Group A. Group B displayed a BMD increase (± SD) at the lumbar spine of +4.0 % (± 2.1) and +5.9 % (± 2.0) at 1 and 2 year(s), respectively, compared with +2.8 % (± 1.7) and +3.5 % (± 2.2), respectively, observed for Group A. Similarly, Group B showed better performance compared with Group A for BMD increase at the femoral neck, with an observed increase of +3.5 % (± 1.7) and +5.4 % (± 1.8) at 1 and 2 year(s), respectively, compared with a change of +2.3 % (± 1.9) and +2.5 % (± 1.9), respectively, registered in Group A. Consistently, the BMD increase measured at the total femur was significantly higher for Group B [+3.4 % (± 1.9) and +4.9 % (± 2.1) at years 1 and 2, respectively] compared with Group A [+1.6 % (± 0.9) and +2.4 % (± 1.9) at years 1 and 2, respectively]. When the change in BMD from year 1 to year 2 was compared, a significant increase of BMD was seen in Group B in all the analysed regions, contrary to that observed for Group A where a plateau effect resulted in no significant change from year 1 to year 2. Three non-traumatic vertebral fractures occurred during the study: two in Group A and one in Group B. The present study indicates the superior performance of IM clodronate 200 mg weekly (100 mg twice weekly) compared with 100 mg once weekly in BMD in women with postmenopausal osteoporosis. This work demonstrated that administration of twice the drug dosage in a week significantly improved the efficacy of the treatment without inducing serious adverse events. Therefore, IM clodronate 200 mg weekly may be considered a valid therapeutic choice for the treatment of postmenopausal osteoporosis.

Aqueous extract from Brownea grandiceps flowers with effect on coagulation and fibrinolytic system.

PubMed

Pereira, Betzabeth; Brazón, Josmary

2015-02-03

Brownea grandiceps flowers are used in Venezuelan folk medicine as anti-hemorrhagic in women with heavy menstrual blood loss (menorrhagia). However, prior to this study, there were no scientific investigations to support this fact, because the aqueous extract from Brownea grandiceps flowers had not been previously evaluated neither phytochemically nor biologically. The objective of this work was to evaluate in vitro the effects of aqueous extract from Brownea grandiceps flowers on the coagulation system and fibrinolysis. An infusion of Brownea grandiceps flowers (160g) was performed; then, it was homogenized, centrifuged and lyophilized to obtain the aqueous extract, and this was called BGE. Subsequently, the extract was characterized on the one hand, phytochemically and on the other hand, biologically, employing prothrombin time (PT), partial thromboplastin time (PTT) and thrombin time (TT) to determine the effects on extrinsic, intrinsic and common coagulation pathways, respectively. In addition to that, the fibrinogenolytic and fibronectinase activity was evaluated by SDS-PAGE using Tris-Tricine system and analyzed by densitometric study utilizing ImageJ program. Also, by using specific chromogenic substrates for Factor Xa (FXa), thrombin, tissue plasminogen activator (t-PA), urokinase plasminogen activator (u-PA) and plasmin, it was assessed whether BGE exhibited some enzyme-like activity, and inhibitory activity of the afore mentioned enzymes. Fibrinolytic and antifibrinolytic activities were determined by a fibrin plate method. Data were analyzed by an nonparametric method. BGE presented tannins, saponins, glycosides, alkaloids, flavonoids, coumarins, and did not contain triterpenoids and steroids. Also, BGE at low concentrations (250-1250µg/mL) reduced the PT, while higher concentrations (15000-25000µg/mL) prolonged this time. However, BGE concentrations between 1250 and 25000µg/mL prolonged the PTT. Prolongation of PT and PTT was observed at high concentrations and was due to FXa inhibitor found in BGE and this effect could be strengthened by degradation of fibrinogen and fibronectin, which were also produced by BGE. Moreover, BGE did not clot fibrinogen or human plasma, and neither did it cleave the chromogenic substrates specific to FXa nor thrombin. These results suggest the pro-coagulant components could be acting on some factor of the extrinsic pathway, since only PT was shortened. Furthermore, BGE did not hydrolyze the chromogenic substrate specific to plasmin, t-PA and u-PA nor did it produce fibrin degradation. However, all BGE concentrations tested inhibited the plasmin activity in a dose-dependent manner. The outcomes of this study reveal the presence of fibrinogenolytic, fibronectinase and anti-FXa components in BGE, plus anti-plasmin compounds that could be acting as antifibrinolytic, thus delaying the fibrin degradation in pathophysiological processes, as it has been observed in women presenting with menorrhagia due to a high plasmin concentration. Where this anti-plasmin compound, along with pro-coagulant components also present in BGE, could be made responsible for reducing heavy menstrual bleeding in women, since a deficiency in one or more blood coagulation factors such as factor VII, V or X, is a potential cause of menorrhagia. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Pigmented villonodular synovitis (PVNS) of the knee joint: magnetic resonance imaging (MRI) using standard and dynamic paramagnetic contrast media. Report of 52 cases surgically and histologically controlled.

PubMed

Barile, Antonio; Sabatini, Mylene; Iannessi, Francesca; Di Cesare, Ernesto; Splendiani, Alessandra; Calvisi, Vittorio; Masciocchi, Carlo

2004-04-01

Pigmented villonodular synovitis (PVNS) is a rare proliferative disorder of the synovial membrane, exhibiting benign behaviour from a biological point of view. This kind of synovial hyperplasia leads to the formation of villi and nodules characterized by deposit of intracellular haemosiderin. It primarily involves young adults, the peak age being between the second and fourth decade of life. It may appear either in a diffuse or a localized (nodular) form. The joint most affected is the knee and diffuse PVNS is the most common form. Diagnostic imaging techniques, particularly MRI, allow lesion identification, suggesting a diagnosis. However, such diagnosis can be confirmed only on histology as the final diagnosis of PVNS, and therefore the possibility of differential diagnosis with other haemorrhagic and chronic hyperplastic synovites, is based on the detection of intracellular haemosiderin components. The aim of this study is to evaluate the usefulness of MRI, which might be completed with the intravenous injection of contrast medium, in the characterization of such pathological picture. From January 1999 to December 2002, we evaluated 52 patients presenting knee swelling, pain and functional impairment. Only 19 patients had a history of trauma. All patients underwent MRI using a dedicated 0.2 T unit or a whole-body' 1.5 T unit. In 30 cases the baseline examination was completed with intravenous injection of contrast medium, followed by dynamic 3D-SPGR sequences at 45, 90, 135 and 225 seconds from the initial injection. These dynamic sequences were then processed by means of early and late subtractions, evaluating the regions of interest (ROI) positioned in the areas with higher post-contrast enhancement. Thirty-eight patients had been previously submitted to Ultrasonography (US), whereas twenty-five patients to Computed Tomography (TC). Later, all patients underwent surgery. Only two patients required an arthrotomy. We then retrospectively evaluated the imaging findings obtained, comparing them with the histological data. Patients affected by autoimmune and systemic inflammatory disorders were excluded from this study. The suspected diagnosis of PVNS was confirmed in 44/52 patients examined. CT examination allowed to detect the presence of a synovial proliferation with densitometric values ranging from 55 to 75 Hounsfield Units (HU) in all cases. In 11 cases, US examination revealed the presence of nodular hyperechoic structures surrounded by anechoic areas, with no differentiation between diffuse and nodular forms. Baseline MRI images showed no differential features among the various histological forms detected. In fact, the nodular structures demonstrated intermediate-to-low intensity signal in all sequences performed. Contrast enhanced MRI showed the presence of areas of inhomogeneous signal due to the increased intensity signal of hypervascular areas. The analysis of vascular dynamics demonstrated a characteristic exponential intensity/time curve both in diffuse and localized forms. The definition of pigmented villonodular synovitis was first employed by Jaffé in 1941 to describe the benign proliferative inflammatory nature of such pathology, characterized by a thickened and hyperplastic synovia organized into villi and nodules, leading to deposition of intracellular haemosiderin pigments. Presently, Authors prefer to include in hemorrhagic synovites all chronic and haemorragic synovial disorders, regardless of the aetiopathogenesis (rheumatoid arthritis, arthropathy secondary to haemorrhagic diathesis, chronic articular traumatism, haemangioma, synovial sarcoma). PVNS involves young adults, with no gender preference; it affects the knee joint in 66-80% of cases, with no typical symptomatology. The absolute absence of any characteristic feature makes a correct differential diagnosis difficult. So far, the only possibility to diagnose PVNS is based on the histological examination: presence of intracellular and subsynovial haemosiderin pigments, predominance of nodular structures as compared to villi, presence of macrophage multinucleate cells, production of collagen, mitotic cellular elements. Therefore, the possibility of characterizing PVNS using MRI is based on detection of a higher number of nodules as compared to villi, as the presence of haemosiderin is always characterized by low signal intensity on T2-weighted images, both intra- and extracellularly. New information on MRI semeiotics has come from the use of post-contrast enhanced dynamic sequences which are able to provide semi-quantitative data on CM velocity distribution within the hyperplastic areas. However, in our experience, dynamic-enhanced MRI did not provide any differential feature between PVNS and the other chronic hemorrhagic forms. Any inflammatory pathology leads to an increased capillary permeability with progressive deposit of CM in the area of interest. In all cases examined, the maximum deposit of contrast medium was observed in the extracellular phase, with a delayed wash-out. PVNS of the knee presents a difficult differential diagnosis. In many cases, only MRI is able to identify the presence of haemosiderin precipitates within the nodules characterizing the lesion. The use of standard and dynamic contrast media seems unable to provide additional diagnostic information. Thus, the diagnosis still pertains to histology.

[The influence of hormonal replacement therapy on bone density in postmenopausal women depending on polymorphism of vitamin D receptor (VDR) and estrogen receptor (ER) genes].

PubMed

Brodowska, Agnieszka

2003-01-01

Osteoporosis is still an important health problem in modern societies. The densitometric criterion for the diagnosis of this condition established by WHO in 1994 is bone mass density (BMD) lower than 2.5 standard deviation (SD) from the mean value for young healthy individuals of the same sex. Between 60 and 90% of bone density (quantity of bone tissue in the human skeleton) at the time when growth is terminated is genetically determined. For this reason, genes predisposing to osteoporosis and mechanisms of their activity remain the object of investigations. Among them are genes coding for vitamin D receptor (VDR), estrogen receptor (ER), type I collagen, TGF-beta and IL-6. Diminishing bone density past the age of thirty is a physiologic process. Bone loss averages 0.3-0.6% per year. Acceleration of this process to 1.2-6% per year in postmenopausal women has been attributed to constantly decreasing estrogen concentration. Hence, the gold standard in osteoporosis prevention and treatment includes estrogen-progestagen therapy enriched with vitamin D analogues, calcium-rich diet and regular physical exercises. Treatment of osteoporosis can be long and expensive. The condition may lead to disability. Osteoporotic fractures and their complications may be fatal. For these reasons, the chief priority in osteoporosis is prevention. Unfortunately, current diagnostic methods (for detection of osteoporosis and monitoring of treatment) remain unsatisfactory. Molecular techniques offer a promising approach to diagnosis and monitoring of therapy. Additionally, the risk of osteoporosis in 1st degree relatives can be assessed and early prevention can be started. The present study addressed the following questions: 1. Are there differences in spine BMD in untreated women with postmenopausal osteoporosis depending on polymorphism of VDR and ER genes? 2. Does efficacy of treatment (increase in spine BMD) in women with postmenopausal osteoporosis depend on polymorphism of VDR and ER genes? 3. What estrogen concentration is necessary to protect bone tissue depending on the polymorphism of VDR and ER genes? The study group included 44 postmenopausal women aged 44-75 years with primary osteoporosis on cyclic HRT (hormonal replacement therapy). Two hormonal preparations were administered: Systen 50 (Jansen Cilag) transdermal system twice per week between day 1 and 21 of the cycle; Provera (Upjohn) 5 mg tablets daily between day 16 and 27 of the cycle. This therapy was supplemented with vitamin D analogue (Alphacalcidolum, Glaxo-Poznan) orally at 0.25 microg per day, calcium-enriched (1200 mg daily) diet and regular physical exercise. Patients were qualified to the study on the basis of a questionnaire. Women with secondary osteoporosis were excluded. TSH, FT3, and FT4 concentrations, as well as fasting glucose were measured. 24 h glycemia was established in women with elevated glucose levels. Polymorphism of the ER gene was studied with Xba I and Pvu II restrictases. Polymorphism of the VDR gene was studied with Bsm I restrictase. Age and BMI were recorded. Spine BMD was determined with DEXA (Dual Energy X-ray Absorptiometry (Lunar instrument) before treatment and after 12 months of HRT. Serum estradiol concentrations were measured before treatment and after 2 months of HTR. The following conclusions were drawn: 1. There is no connection between VDR and ER gene polymorphism and degree of osteoporosis before treatment. 2. XX, PP and Bb markers or X, P, B alleles are associated with a significant decrease in therapeutic efficacy. Nevertheless, satisfactory results were achieved in each woman with primary osteoporosis. 3. Estradiol concentration in serum before and during HRT does not depend on the polymorphism of VDR and ER genes.

A New Method for the Determination of Annual Sediment Fluxes from Varved Lake Sediments

NASA Astrophysics Data System (ADS)

Francus, P.; Massa, C.; Lapointe, F.

2013-12-01

Calculation of sediment mass accumulation rates instead of thickness accumulation is preferable for paleoclimatic reconstruction as it eliminates the effects of dilution and compaction. Annually laminated lake sediment sequences (varved) theoretically allow for the estimation of sediment fluxes at annual scale, but the calculation is limited by discrete bulk density measurements, often carried out at a much lower resolution (usually 1 cm) than the varves (ranging from 0.07 to 27.3 mm, average 1.84 mm according to Ojala et al. 2012). Since many years the development of automated logging instruments made available continuous and high resolution sediment property data, in a non-destructive fashion. These techniques can easily be used to extract the physical and chemical parameters of sediments at the varve scale (down to 100 μm). Here we present a robust method to calculate annual sediment fluxes from varved lake sediments by combining varves thickness measurements to core logging data, and provide an example for its applications. Several non-destructive densitometric methods applied to the Strathcona Lake sediment, northern Ellesmere Island, Canada (78°33'N; 82°05'W) were compared: Hounsfield Units from a CT-Scan, coherent/incoherent ratio and X-ray radiography (of both split core and sediment slabs, from an Itrax core Scanner), and gamma ray attenuation density. Core logging data were statistically compared to 400 discrete measurements of dry bulk density, wet bulk density and water content performed at 2 mm contiguous intervals. A very strong relationship was found between X-ray grey level on sediment slab and dry bulk density. Relative X-ray densities, at 100μm resolution, were then successfully calibrated against real densities. The final step consisted in binning the calibrated densities to the corresponding varve thickness and then to calculate the annual mass accumulation rates by multiplying the two parameters for each varve year. Strathcona Lake is located directly downstream of the Agassiz ice cap and contains laminated sediments whose accumulation is directly related to hydrological inputs generated by the melting of the ice cap. Over the last 65 years, annual sediment accumulation rates in Strathcona Lake documented an increase in high-energy hydrologic discharge events from 1990 to 2009. This timing is in agreement with evidence for an increase in the amount of melt on the adjacent Agassiz Ice Cap, as recorded in ice cores. A good correspondence was also found between annual mass accumulation rates and Eureka air temperature records, suggesting that temperature changes affected the extent of summer melting on the Agassiz Ice Cap, leading to high sediment yield to Strathcona Lake. Ojala, A.E.K., Francus, P., Zolitschka, B., Besonen, M. and Lamoureux, S.F. (2012) Characteristics of sedimentary varve chronologies - A review. Quaternary Science Reviews, 43, 45-60.

Value of two-phase dynamic multidetector computed tomography in differential diagnosis of post-inflammatory strictures from esophageal cancer

PubMed Central

Karmazanovsky, Grigory G; Buryakina, Svetlana A; Kondratiev, Evgeny V; Yang, Qin; Ruchkin, Dmitry V; Kalinin, Dmitry V

2015-01-01

AIM: To characterize the computed tomography (CT) findings in patients with post-inflammatory esophageal strictures (corrosive and peptic) and reveal the optimal scanning phase protocols for distinguishing post-inflammatory esophageal stricture and esophageal cancer. METHODS: Sixty-five patients with esophageal strictures of different etiology were included in this study: 24 patients with 27 histopathologically confirmed corrosive strictures, 10 patients with 12 peptic strictures and 31 patients with esophageal cancer were evaluated with a two-phase dynamic contrast-enhanced MDCT. Arterial and venous phases at 10 and 35 s after the attenuation of 200 HU were obtained at the descending aorta, with a delayed phase at 6-8 min after the start of injection of contrast media. For qualitative analysis, CT scans of benign strictures were reviewed for the presence/absence of the following features: “target sign”, luminal mass, homogeneity of contrast medium uptake, concentric wall thickening, conically shaped suprastenotic dilatation, smooth boundaries of stenosis and smooth mucous membrane at the transition to stenosis, which were compared with a control group of 31 patients who had esophageal cancer. The quantitative analysis included densitometric parameter acquisition using regions-of-interest measurement of the zone of stenosis and normal esophageal wall and the difference between those measurements (ΔCT) at all phases of bolus contrast enhancement. Esophageal wall thickening, length of esophageal wall thickening and size of the regional lymph nodes were also evaluated. RESULTS: The presence of a concentric esophageal wall, conically shaped suprastenotic dilatation, smooth upper and lower boundaries, “target sign” and smooth mucous membrane at the transition to stenosis were suggestive of a benign cause, with sensitivities of 92.31%, 87.17%, 94.87%, 76.92% and 82.05%, respectively, and specificities of 70.96%, 89.66%, 80.65%, 96.77% and 93.55%, respectively. The features that were most suggestive of a malignant cause were eccentric esophageal wall thickening, tuberous upper and lower boundaries of stenosis, absence of mucous membrane visualization, rupture of the mucous membrane at the upper boundary of stenosis, cup-shaped suprastenotic dilatation, luminal mass and enlarged regional lymph nodes with specificities of 92.31% 94.87%, 67.86%, 100%, 97.44%, 94.87% and 82.86%, respectively and sensitivities of 70.97%, 80.65%, 96.77%, 80.65%, 54.84%, 87.10% and 60%, respectively. The highest tumor attenuation occurred in the arterial phase (mean attenuation 74.13 ± 17.42 HU), and the mean attenuation difference between the tumor and the normal esophageal wall (mean ΔCT) in the arterial phase was 23.86 ± 19.31 HU. Here, 11.5 HU of ΔCT in the arterial phase was the cut-off value used to differentiate esophageal cancer from post-inflammatory stricture (P = 0.000). The highest attenuation of post-inflammatory strictures occurred in the delayed phase (mean attenuation 71.66 ± 14.28 HU), and the mean ΔCT in delayed phase was 34.03 ± 15.94 HU. Here, 18.5 HU of ΔCT in delayed phase was the cut-off value used to differentiate post-inflammatory stricture from esophageal cancer (P < 0.0001). CONCLUSION: The described imaging findings reveal high diagnostic significance in the differentiation of benign strictures from esophageal cancer. PMID:26269677

A Thematic Review of Studies into the Effectiveness of Context-Based Chemistry Curricula

ERIC Educational Resources Information Center

Ultay, Neslihan; Calik, Muammer

2012-01-01

Context-based chemistry education aims at making connections between real life and the scientific content of chemistry courses. The purpose of this study was to evaluate context-based chemistry studies. In looking for the context-based chemistry studies, the authors entered the keywords "context-based", "contextual learning" and "chemistry…

Sensor Technology Baseline Study for Enabling Condition Based Maintenance Plus in Army Ground Vehicles

DTIC Science & Technology

2012-03-01

for enabling condition based maintenance plus in Army ground vehicles. The sensor study was driven from Failure Mode Effects Analysis ( FMEA ...of Tables Table 1. Sensor technology baseline study based on engine FMEA report. ...................................5 Table 2. Sensor technology...baseline study based on transmission FMEA report. .........................8 Table 3. Sensor technology baseline study based on alternator FMEA report

A narrative review of undergraduate peer-based healthcare ethics teaching.

PubMed

Hindmarch, Thomas; Allikmets, Silvia; Knights, Felicity

2015-12-12

This study explores the literature in establishing the value of undergraduate peer-based healthcare ethics teaching as an educational methodology. A narrative review of the literature concerning peer-based ethics teaching was conducted. MEDLINE, EMBASE, CINAHL, SCOPUS databases, and the Cochrane Library, were systematically searched for studies of peer-based ethics or professionalism teaching. Selected studies related peer-based teaching to ethics education outcomes. Ten publications were identified. Selected studies were varied in their chosen intervention methodology and analysis. Collectively, the identified studies suggest peer-based ethics education is an effective and valued educational methodology in training healthcare professionals. One paper suggests peer-based ethics teaching has advantages over traditional didactic methods. Peer-based ethics teaching also receives positive feedback from student participants. However, the limited literature base demonstrates a clear need for more evaluation of this pedagogy. The current literature base suggests that undergraduate peer based healthcare ethics teaching is valuable in terms of efficacy and student satisfaction. We conclude that the medical community should invest in further study in order to capitalise upon the potential of peer-based ethics teaching in undergraduate healthcare education.

Does the use of efficacy or effectiveness evidence in cost-effectiveness analysis matter?

PubMed

Dilokthornsakul, Piyameth; Chaiyakunapruk, Nathorn; Campbell, Jonathan D

2017-01-02

To test the association of clinical evidence type, efficacy-based or effectiveness-based ("E"), versus whether or not asthma interventions' cost-effectiveness findings are favorable. We conducted a systematic review of PubMed, EMBASE, Tufts CEA registry, Cochrane CENTRAL, and the UK National Health Services Economic Evaluation Database from 2009 to 2014. All cost-effectiveness studies evaluating asthma medication(s) were included. Clinical evidence type, "E," was classified as efficacy-based if the evidence was from an explanatory randomized controlled trial(s) or meta-analysis, while evidence from pragmatic trial(s) or observational study(s) was classified as effectiveness-based. We defined three times the World Health Organization cost-effectiveness willingness-to-pay (WTP) threshold or less as a favorable cost-effectiveness finding. Logistic regression tested the likelihood of favorable versus unfavorable cost-effectiveness findings against the type of "E." 25 cost-effectiveness studies were included. Ten (40.0%) studies were effectiveness-based, yet 15 (60.0%) studies were efficacy-based. Of 17 studies using endpoints that could be compared to WTP threshold, 7 out of 8 (87.5%) effectiveness-based studies yielded favorable cost-effectiveness results, whereas 4 out of 9 (44.4%) efficacy-based studies yielded favorable cost-effectiveness results. The adjusted odds ratio was 15.12 (95% confidence interval; 0.59 to 388.75) for effectiveness-based versus efficacy-based achieving favorable cost-effectiveness findings. More asthma cost-effectiveness studies used efficacy-based evidence. Studies using effectiveness-based evidence trended toward being more likely to disseminate favorable cost-effective findings than those using efficacy. Health policy decision makers should pay attention to the type of clinical evidence used in cost-effectiveness studies for accurate interpretation and application.

Systematic review of skills transfer after surgical simulation-based training.

PubMed

Dawe, S R; Pena, G N; Windsor, J A; Broeders, J A J L; Cregan, P C; Hewett, P J; Maddern, G J

2014-08-01

Simulation-based training assumes that skills are directly transferable to the patient-based setting, but few studies have correlated simulated performance with surgical performance. A systematic search strategy was undertaken to find studies published since the last systematic review, published in 2007. Inclusion of articles was determined using a predetermined protocol, independent assessment by two reviewers and a final consensus decision. Studies that reported on the use of surgical simulation-based training and assessed the transferability of the acquired skills to a patient-based setting were included. Twenty-seven randomized clinical trials and seven non-randomized comparative studies were included. Fourteen studies investigated laparoscopic procedures, 13 endoscopic procedures and seven other procedures. These studies provided strong evidence that participants who reached proficiency in simulation-based training performed better in the patient-based setting than their counterparts who did not have simulation-based training. Simulation-based training was equally as effective as patient-based training for colonoscopy, laparoscopic camera navigation and endoscopic sinus surgery in the patient-based setting. These studies strengthen the evidence that simulation-based training, as part of a structured programme and incorporating predetermined proficiency levels, results in skills transfer to the operative setting. © 2014 BJS Society Ltd. Published by John Wiley & Sons Ltd.

The state of readiness for evidence-based practice among nurses: An integrative review.

PubMed

Saunders, Hannele; Vehviläinen-Julkunen, Katri

2016-04-01

To review factors related to nurses' individual readiness for evidence-based practice and to determine the current state of nurses' evidence-based practice competencies. An integrative review study. Thirty-seven (37) primary research studies on nurses' readiness for evidence-based practice, of which 30 were descriptive cross-sectional surveys, 5 were pretest-posttest studies, and one study each was an experimental pilot study and a descriptive qualitative study. Included studies were published from the beginning of 2004 through end of January 2015. The integrative review study used thematic synthesis, in which the quantitative studies were analyzed deductively and the qualitative studies inductively. Outcomes related to nurses' readiness for evidence-based practice were grouped according to the four main themes that emerged from the thematic synthesis: (1) nurses' familiarity with evidence-based practice (EBP); (2) nurses' attitudes toward and beliefs about evidence-based practice; (3) nurses' evidence-based practice knowledge and skills; and (4) nurses' use of research in practice. Methodological quality of the included studies was evaluated with Joanna Briggs Institute critical appraisal tools. Although nurses were familiar with, had positive attitudes toward, and believed in the value of EBP in improving care quality and patient outcomes, they perceived their own evidence-based practice knowledge and skills insufficient for employing evidence-based practice, and did not use best evidence in practice. The vast majority (81%) of included studies were descriptive cross-sectional surveys, 84% used a non-probability sampling method, sample sizes were small, and response rates low. Most included studies were of modest quality. More robust, theoretically-based and psychometrically sound nursing research studies are needed to test and evaluate the effectiveness of interventions designed to advance nurses' evidence-based practice competencies, especially teaching them how to integrate evidence-based practice into clinical decision-making. All efforts should be focused on systematically using knowledge transformation strategies shown to be effective in rigorous studies, to translate best evidence into practice-friendly, readily usable forms that are easily accessible to nurses to integrate into their clinical practice. Copyright © 2015 Elsevier Ltd. All rights reserved.

A narrative review of undergraduate peer-based healthcare ethics teaching

PubMed Central

Allikmets, Silvia; Knights, Felicity

2015-01-01

Objectives This study explores the literature in establishing the value of undergraduate peer-based healthcare ethics teaching as an educational methodology. Methods A narrative review of the literature concerning peer-based ethics teaching was conducted. MEDLINE, EMBASE, CINAHL, SCOPUS databases, and the Cochrane Library, were systematically searched for studies of peer-based ethics or professionalism teaching. Selected studies related peer-based teaching to ethics education outcomes. Results Ten publications were identified. Selected studies were varied in their chosen intervention methodology and analysis. Collectively, the identified studies suggest peer-based ethics education is an effective and valued educational methodology in training healthcare professionals. One paper suggests peer-based ethics teaching has advantages over traditional didactic methods. Peer-based ethics teaching also receives positive feedback from student participants. However, the limited literature base demonstrates a clear need for more evaluation of this pedagogy. Conclusions The current literature base suggests that undergraduate peer based healthcare ethics teaching is valuable in terms of efficacy and student satisfaction. We conclude that the medical community should invest in further study in order to capitalise upon the potential of peer-based ethics teaching in undergraduate healthcare education. PMID:26668050

A Thematic Review of Studies into the Effectiveness of Context-Based Chemistry Curricula

NASA Astrophysics Data System (ADS)

Ültay, Neslihan; Çalık, Muammer

2012-12-01

Context-based chemistry education aims at making connections between real life and the scientific content of chemistry courses. The purpose of this study was to evaluate context-based chemistry studies. In looking for the context-based chemistry studies, the authors entered the keywords `context-based', `contextual learning' and `chemistry education' in well-known databases (i.e. Academic Search Complete, Education Research Complete, ERIC, Springer LINK Contemporary). Further, in case the computer search by key words may have missed a rather substantial part of the important literature in the area, the authors also conducted a hand search of the related journals. To present a detailed thematic review of context-based chemistry studies, a matrix was used to summarize the findings by focusing on insights derived from the related studies. The matrix incorporates the following themes: needs, aims, methodologies, general knowledge claims, and implications for teaching and learning, implications for curriculum development and suggestions for future research. The general knowledge claims investigated in this paper were: (a) positive effects of the context-based chemistry studies; (b) caveats, both are examined in terms of students' attitudes and students' understanding/cognition. Implications were investigated for practice in context- based chemistry studies, for future research in context- based chemistry studies, and for curriculum developers in context- based chemistry studies. Teachers of context-based courses claimed that the application of the context-based learning approach in chemistry education improved students' motivation and interest in the subject. This seems to have generated an increase in the number of the students who wish to continue chemistry education at higher levels. However, despite the fact that the majority of the studies have reported advantages of context-based chemistry studies, some of them have also referred to pitfalls, i.e. dominant structure of out-of-school learning, tough nature of some chemistry topics, and teacher anxiety of lower-ability students.

Effect of Web-Based Versus Paper-Based Questionnaires and Follow-Up Strategies on Participation Rates of Dutch Childhood Cancer Survivors: A Randomized Controlled Trial

PubMed Central

van Dulmen-den Broeder, Eline; van der Pal, Helena J; Hollema, Nynke; Kremer, Leontien C; van den Heuvel-Eibrink, Marry M; van Leeuwen, Flora E

2015-01-01

Background Questionnaires are widely used in survey research, especially in cohort studies. However, participation in questionnaire studies has been declining over the past decades. Because high participation rates are needed to limit the risk of selection bias and produce valid results, it is important to investigate invitation strategies which may improve participation. Objectives The purpose of this study is to investigate the effect of Web-based versus paper-based questionnaires on participation rates in a questionnaire survey on late effects among childhood cancer survivors (CCSs). Methods A total of 750 CCSs were randomized across 3 study arms. The initial invitation in study arms 1 and 2 consisted of a Web-based questionnaire only, whereas in study arm 3 this invitation was complemented with a paper-based version of the questionnaire. The first postal reminder, sent to the nonresponding CCSs in all 3 study arms, consisted of either a reminder letter only (study arms 1 and 3) or a reminder letter complemented with a paper-based questionnaire (study arm 2). The second postal reminder was restricted to CCSs in study arms 1 and 2, with only those in study arm 1 also receiving a paper-based questionnaire. CCSs in study arm 3 received a second reminder by telephone instead of by mail. In contrast to CCSs in study arm 3, CCSs in study arms 1 and 2 received a third reminder, this time by telephone. Results: Overall, 58.1% (436/750) of the CCSs participated in the survey. Participation rates were equal in all 3 study arms with 57.4% (143/249) in arm 1, 60.6% (152/251) in arm 2, and 56.4% (141/250) in arm 3 (P=.09). Participation rates of CCSs who received an initial invitation for the Web-based questionnaire only and CCSs who received an invitation to complete either a paper-based or Web-based questionnaire did not differ (P=.55). After the first postal reminder, participation rates of CCSs invited for the Web-based questionnaire only also did not differ compared with CCSs invited for both the Web-based and paper-based questionnaires (P=.48). In general, CCSs preferred the paper-based over the Web-based questionnaire, and those completing the paper-based questionnaire were more often unemployed (P=.004) and lower educated (P<.001). Conclusion Invitation strategies offering a Web-based questionnaire without a paper-based alternative at first invitation can be used without compromising participation rates of CCS. Offering the choice between paper- and Web-based questionnaires seems to result in the highest accrual participation rate. Future research should look into the quality of the data delivered by both questionnaires filled in by respondents themselves. Trial Registration International Standard Randomized Controlled Trial Number (ISRCTN): 84711754; http://www.controlled-trials.com/ISRCTN84711754 (Archived by WebCite at http://www.webcitation.org/6c9ZB8paX) PMID:28410161

A systematic review of surgical skills transfer after simulation-based training: laparoscopic cholecystectomy and endoscopy.

PubMed

Dawe, Susan R; Windsor, John A; Broeders, Joris A J L; Cregan, Patrick C; Hewett, Peter J; Maddern, Guy J

2014-02-01

A systematic review to determine whether skills acquired through simulation-based training transfer to the operating room for the procedures of laparoscopic cholecystectomy and endoscopy. Simulation-based training assumes that skills are directly transferable to the operation room, but only a few studies have investigated the effect of simulation-based training on surgical performance. A systematic search strategy that was used in 2006 was updated to retrieve relevant studies. Inclusion of articles was determined using a predetermined protocol, independent assessment by 2 reviewers, and a final consensus decision. Seventeen randomized controlled trials and 3 nonrandomized comparative studies were included in this review. In most cases, simulation-based training was in addition to patient-based training programs. Only 2 studies directly compared simulation-based training in isolation with patient-based training. For laparoscopic cholecystectomy (n = 10 studies) and endoscopy (n = 10 studies), participants who reached simulation-based skills proficiency before undergoing patient-based assessment performed with higher global assessment scores and fewer errors in the operating room than their counterparts who did not receive simulation training. Not all parameters measured were improved. Two of the endoscopic studies compared simulation-based training in isolation with patient-based training with different results: for sigmoidoscopy, patient-based training was more effective, whereas for colonoscopy, simulation-based training was equally effective. Skills acquired by simulation-based training seem to be transferable to the operative setting for laparoscopic cholecystectomy and endoscopy. Future research will strengthen these conclusions by evaluating predetermined competency levels on the same simulators and using objective validated global rating scales to measure operative performance.

Making a Case for a Blended Approach: The Need for The Design-Based Case Study

ERIC Educational Resources Information Center

Deaton, Cynthia C. M.; Malloy, Jacquelynn A.

2017-01-01

Design-based case studies address research questions that involve instructional innovations within a bounded system. This blend of case study and design-based research provides a systematic approach to examining instructional innovations that are bounded by perspective, context, and time. Design-based case studies provide a framework for engaging…

Teacher Implementation of Reform-Based Mathematics and Implications for Algebra Readiness: A Qualitative Study of 4th Grade Classrooms

ERIC Educational Resources Information Center

Sher, Stephen Korb

2011-01-01

This study looked at 4th grade classrooms to see "how" teachers implement NCTM standards-based or reform-based mathematics instruction and then analyzed it for the capacity to improve students' "algebra readiness." The qualitative study was based on classroom observations, teacher and administrator interviews, and teacher surveys. The study took…

Report on Pairing-based Cryptography.

PubMed

Moody, Dustin; Peralta, Rene; Perlner, Ray; Regenscheid, Andrew; Roginsky, Allen; Chen, Lily

2015-01-01

This report summarizes study results on pairing-based cryptography. The main purpose of the study is to form NIST's position on standardizing and recommending pairing-based cryptography schemes currently published in research literature and standardized in other standard bodies. The report reviews the mathematical background of pairings. This includes topics such as pairing-friendly elliptic curves and how to compute various pairings. It includes a brief introduction to existing identity-based encryption (IBE) schemes and other cryptographic schemes using pairing technology. The report provides a complete study of the current status of standard activities on pairing-based cryptographic schemes. It explores different application scenarios for pairing-based cryptography schemes. As an important aspect of adopting pairing-based schemes, the report also considers the challenges inherent in validation testing of cryptographic algorithms and modules. Based on the study, the report suggests an approach for including pairing-based cryptography schemes in the NIST cryptographic toolkit. The report also outlines several questions that will require further study if this approach is followed.

Report on Pairing-based Cryptography

PubMed Central

Moody, Dustin; Peralta, Rene; Perlner, Ray; Regenscheid, Andrew; Roginsky, Allen; Chen, Lily

2015-01-01

This report summarizes study results on pairing-based cryptography. The main purpose of the study is to form NIST’s position on standardizing and recommending pairing-based cryptography schemes currently published in research literature and standardized in other standard bodies. The report reviews the mathematical background of pairings. This includes topics such as pairing-friendly elliptic curves and how to compute various pairings. It includes a brief introduction to existing identity-based encryption (IBE) schemes and other cryptographic schemes using pairing technology. The report provides a complete study of the current status of standard activities on pairing-based cryptographic schemes. It explores different application scenarios for pairing-based cryptography schemes. As an important aspect of adopting pairing-based schemes, the report also considers the challenges inherent in validation testing of cryptographic algorithms and modules. Based on the study, the report suggests an approach for including pairing-based cryptography schemes in the NIST cryptographic toolkit. The report also outlines several questions that will require further study if this approach is followed. PMID:26958435

Journal Quality in Mathematics Education

ERIC Educational Resources Information Center

Williams, Steven R.; Leatham, Keith R.

2017-01-01

We present the results of 2 studies, a citation-based study and an opinion-based study, that ranked the relative quality of 20 English-language journals that exclusively or extensively publish mathematics education research. We further disaggregate the opinion-based data to provide insights into variations in judgment of journal quality based on…

A Collective Case Study of Secondary Students' Model-Based Inquiry on Natural Selection through Programming in an Agent-Based Modeling Environment

ERIC Educational Resources Information Center

Xiang, Lin

2011-01-01

This is a collective case study seeking to develop detailed descriptions of how programming an agent-based simulation influences a group of 8th grade students' model-based inquiry (MBI) by examining students' agent-based programmable modeling (ABPM) processes and the learning outcomes. The context of the present study was a biology unit on…

Contests versus Norms: Implications of Contest-Based and Norm-Based Intervention Techniques

PubMed Central

Bergquist, Magnus; Nilsson, Andreas; Hansla, André

2017-01-01

Interventions using either contests or norms can promote environmental behavioral change. Yet research on the implications of contest-based and norm-based interventions is lacking. Based on Goal-framing theory, we suggest that a contest-based intervention frames a gain goal promoting intensive but instrumental behavioral engagement. In contrast, the norm-based intervention was expected to frame a normative goal activating normative obligations for targeted and non-targeted behavior and motivation to engage in pro-environmental behaviors in the future. In two studies participants (n = 347) were randomly assigned to either a contest- or a norm-based intervention technique. Participants in the contest showed more intensive engagement in both studies. Participants in the norm-based intervention tended to report higher intentions for future energy conservation (Study 1) and higher personal norms for non-targeted pro-environmental behaviors (Study 2). These findings suggest that contest-based intervention technique frames a gain goal, while norm-based intervention frames a normative goal. PMID:29218026

Contests versus Norms: Implications of Contest-Based and Norm-Based Intervention Techniques.

PubMed

Bergquist, Magnus; Nilsson, Andreas; Hansla, André

2017-01-01

Interventions using either contests or norms can promote environmental behavioral change. Yet research on the implications of contest-based and norm-based interventions is lacking. Based on Goal-framing theory, we suggest that a contest-based intervention frames a gain goal promoting intensive but instrumental behavioral engagement. In contrast, the norm-based intervention was expected to frame a normative goal activating normative obligations for targeted and non-targeted behavior and motivation to engage in pro-environmental behaviors in the future. In two studies participants ( n = 347) were randomly assigned to either a contest- or a norm-based intervention technique. Participants in the contest showed more intensive engagement in both studies. Participants in the norm-based intervention tended to report higher intentions for future energy conservation (Study 1) and higher personal norms for non-targeted pro-environmental behaviors (Study 2). These findings suggest that contest-based intervention technique frames a gain goal, while norm-based intervention frames a normative goal.

Adapting practice-based intervention research to electronic environments: opportunities and complexities at two institutions.

PubMed

Stille, Christopher J; Lockhart, Steven A; Maertens, Julie A; Madden, Christi A; Darden, Paul M

2015-01-01

Primary care practice-based research has become more complex with increased use of electronic health records (EHRs). Little has been reported about changes in study planning and execution that are required as practices change from paper-based to electronic-based environments. We describe the evolution of a pediatric practice-based intervention study as it was adapted for use in the electronic environment, to enable other practice-based researchers to plan efficient, effective studies. We adapted a paper-based pediatric office-level intervention to enhance parent-provider communication about subspecialty referrals for use in two practice-based research networks (PBRNs) with partially and fully electronic environments. We documented the process of adaptation and its effect on study feasibility and efficiency, resource use, and administrative and regulatory complexities, as the study was implemented in the two networks. Considerable time and money was required to adapt the paper-based study to the electronic environment, requiring extra meetings with institutional EHR-, regulatory-, and administrative teams, and increased practice training. Institutional unfamiliarity with using EHRs in practice-based research, and the consequent need to develop new policies, were major contributors to delays. Adapting intervention tools to the EHR and minimizing practice disruptions was challenging, but resulted in several efficiencies as compared with a paper-based project. In particular, recruitment and tracking of subjects and data collection were easier and more efficient. Practice-based intervention research in an electronic environment adds considerable cost and time at the outset of a study, especially for centers unfamiliar with such research. Efficiencies generated have the potential of easing the work of study enrollment, subject tracking, and data collection.

Team-Based Care with Pharmacists to Improve Blood Pressure: a Review of Recent Literature.

PubMed

Kennelty, Korey A; Polgreen, Linnea A; Carter, Barry L

2018-01-18

We review studies published since 2014 that examined team-based care strategies and involved pharmacists to improve blood pressure (BP). We then discuss opportunities and challenges to sustainment of team-based care models in primary care clinics. Multiple studies presented in this review have demonstrated that team-based care including pharmacists can improve BP management. Studies highlighted the cost-effectiveness of a team-based pharmacy intervention for BP control in primary care clinics. Little information was found on factors influencing sustainability of team-based care interventions to improve BP control. Future work is needed to determine the best populations to target with team-based BP programs and how to implement team-based approaches utilizing pharmacists in diverse clinical settings. Future studies need to not only identify unmet clinical needs but also address reimbursement issues and stakeholder engagement that may impact sustainment of team-based care interventions.

Online Learning: A Comparison of Web-Based and Land-Based Courses

ERIC Educational Resources Information Center

Brown, Joy L. M.

2012-01-01

Distance learning has become more popular in recent years. Due to concern about the quality of web-based courses, the purpose of this study was to explore the differences in web-based versus land-based courses. In this study, the researcher compares web-based and land-based education courses to explore the strengths and weaknesses of each type of…

Assessment of School-Based Management. [Volume I: Findings and Conclusions.] Studies of Education Reform.

ERIC Educational Resources Information Center

Wohlstetter, Priscilla; Mohrman, Susan Albers

This document presents findings of the Assessment of School-Based Management Study, which identified the conditions in schools that promote high performance through school-based management (SBM). The study's conceptual framework was based on Edward E. Lawler's (1986) model. The high-involvement framework posits that four resources must spread…

Effect of environmental exposure to hydrogen sulfide on central nervous system and respiratory function: a systematic review of human studies

PubMed Central

Lim, Eunjung; Mbowe, Omar; Lee, Angela S. W.; Davis, James

2016-01-01

Background Assessment of the health effects of low-level exposure to hydrogen sulfide (H2S) on humans through experiments, industrial, and community studies has shown inconsistent results. Objective To critically appraise available studies investigating the effect of H2S on the central nervous system (CNS) and on respiratory function. Methods A search was conducted in 16 databases for articles published between January 1980 and July 2014. Two researchers independently evaluated potentially relevant papers based on a set of inclusion/exclusion criteria. Results Twenty-seven articles met the inclusion criteria: 6 experimental, 12 industry-based studies, and 10 community-based studies (one article included both experimental and industry-based studies). The results of the systematic review varied by study setting and quality. Several community-based studies reported associations between day-to-day variations in H2S levels and health outcomes among patients with chronic respiratory conditions. However, evidence from the largest and better-designed community-based studies did not support that chronic, ambient H2S exposure has health effects on the CNS or respiratory function. Results from industry-based studies varied, reflecting the diversity of settings and the broad range of H2S exposures. Most studies did not have individual measurements of H2S exposure. Discussion The results across studies were inconsistent, justifying the need for further research. PMID:27128692

Effect of environmental exposure to hydrogen sulfide on central nervous system and respiratory function: a systematic review of human studies.

PubMed

Lim, Eunjung; Mbowe, Omar; Lee, Angela S W; Davis, James

2016-01-01

Assessment of the health effects of low-level exposure to hydrogen sulfide (H2S) on humans through experiments, industrial, and community studies has shown inconsistent results. To critically appraise available studies investigating the effect of H2S on the central nervous system (CNS) and on respiratory function. A search was conducted in 16 databases for articles published between January 1980 and July 2014. Two researchers independently evaluated potentially relevant papers based on a set of inclusion/exclusion criteria. Twenty-seven articles met the inclusion criteria: 6 experimental, 12 industry-based studies, and 10 community-based studies (one article included both experimental and industry-based studies). The results of the systematic review varied by study setting and quality. Several community-based studies reported associations between day-to-day variations in H2S levels and health outcomes among patients with chronic respiratory conditions. However, evidence from the largest and better-designed community-based studies did not support that chronic, ambient H2S exposure has health effects on the CNS or respiratory function. Results from industry-based studies varied, reflecting the diversity of settings and the broad range of H2S exposures. Most studies did not have individual measurements of H2S exposure. The results across studies were inconsistent, justifying the need for further research.

Moral empiricism and the bias for act-based rules.

PubMed

Ayars, Alisabeth; Nichols, Shaun

2017-10-01

Previous studies on rule learning show a bias in favor of act-based rules, which prohibit intentionally producing an outcome but not merely allowing the outcome. Nichols, Kumar, Lopez, Ayars, and Chan (2016) found that exposure to a single sample violation in which an agent intentionally causes the outcome was sufficient for participants to infer that the rule was act-based. One explanation is that people have an innate bias to think rules are act-based. We suggest an alternative empiricist account: since most rules that people learn are act-based, people form an overhypothesis (Goodman, 1955) that rules are typically act-based. We report three studies that indicate that people can use information about violations to form overhypotheses about rules. In study 1, participants learned either three "consequence-based" rules that prohibited allowing an outcome or three "act-based" rules that prohibiting producing the outcome; in a subsequent learning task, we found that participants who had learned three consequence-based rules were more likely to think that the new rule prohibited allowing an outcome. In study 2, we presented participants with either 1 consequence-based rule or 3 consequence-based rules, and we found that those exposed to 3 such rules were more likely to think that a new rule was also consequence based. Thus, in both studies, it seems that learning 3 consequence-based rules generates an overhypothesis to expect new rules to be consequence-based. In a final study, we used a more subtle manipulation. We exposed participants to examples act-based or accident-based (strict liability) laws and then had them learn a novel rule. We found that participants who were exposed to the accident-based laws were more likely to think a new rule was accident-based. The fact that participants' bias for act-based rules can be shaped by evidence from other rules supports the idea that the bias for act-based rules might be acquired as an overhypothesis from the preponderance of act-based rules. Copyright © 2017 Elsevier B.V. All rights reserved.

Comprehension-Based versus Production-Based Grammar Instruction: A Meta-Analysis of Comparative Studies

ERIC Educational Resources Information Center

Shintani, Natsuko; Li, Shaofeng; Ellis, Rod

2013-01-01

This article reports a meta-analysis of studies that investigated the relative effectiveness of comprehension-based instruction (CBI) and production-based instruction (PBI). The meta-analysis only included studies that featured a direct comparison of CBI and PBI in order to ensure methodological and statistical robustness. A total of 35 research…

An Electronic Library-Based Learning Environment for Supporting Web-Based Problem-Solving Activities

ERIC Educational Resources Information Center

Tsai, Pei-Shan; Hwang, Gwo-Jen; Tsai, Chin-Chung; Hung, Chun-Ming; Huang, Iwen

2012-01-01

This study aims to develop an electronic library-based learning environment to support teachers in developing web-based problem-solving activities and analyzing the online problem-solving behaviors of students. Two experiments were performed in this study. In study 1, an experiment on 103 elementary and high school teachers (the learning activity…

Integrating the Problem-Based Learning Approach into a Web-Based English Reading Course

ERIC Educational Resources Information Center

Lin, Lu-Fang

2018-01-01

This study incorporated the problem-based learning (PBL) approach into a web-based English reading course. The purpose of the study was to examine the impact of PBL on the participants' English reading comprehension (RC) and to explore the participants' perceptions of PBL. The present study conducted a comparison of two teaching methods. Two…

An Exploration of Alternative Scoring Methods Using Curriculum-Based Measurement in Early Writing

ERIC Educational Resources Information Center

Allen, Abigail A.; Poch, Apryl L.; Lembke, Erica S.

2018-01-01

This manuscript describes two empirical studies of alternative scoring procedures used with curriculum-based measurement in writing (CBM-W). Study 1 explored the technical adequacy of a trait-based rubric in first grade. Study 2 explored the technical adequacy of a trait-based rubric, production-dependent, and production-independent scores in…

Bringing antiretroviral therapy (ART) closer to the end-user through mobile clinics and home-based ART: systematic review shows more evidence on the effectiveness and cost effectiveness is needed.

PubMed

Mdege, Noreen Dadirai; Chindove, Stanley

2014-01-01

Home-based antiretroviral therapy (ART) and ART through mobile clinics can potentially increase access to ART for large numbers of people, including hard-to-reach populations. We reviewed literature on the effectiveness and cost implications of the home-based ART and mobile clinic ART models. We searched Medline, Embase, PsycInfo, CINAHL, Cochrane Library, Web of Knowledge and Current Controlled Trials Register for articles published up to March 2012. We included non-randomised and randomised controlled clinical trials that recruited HIV/AIDS positive adults with or without prior exposure to ART. Six studies were included in the review, with only four effectiveness studies (all evaluating home-based ART and none for mobile clinic ART) and four studies reporting on the cost implications. The evidence suggests home-based ART is as effective as health facility-based ART, including on clinical outcomes, viral load and CD4+ count. However, three of these studies were very small. Studies suggest health facility-based ART is the most cost-effective, followed by mobile-clinic ART, with home-based ART being the least cost-effective. Evidence on the effectiveness and cost implications of mobile clinic and home-based ART is currently limited. Although the few available studies suggest home-based ART can potentially be as effective as health facility-based ART, there is need for more research before robust conclusions can be made. Results from the few available studies also suggest that health facility-based ART is the most cost-effective. Copyright © 2013 John Wiley & Sons, Ltd.

Students' Perceptions of Study Modes

ERIC Educational Resources Information Center

Hagel, Pauline; Shaw, Robin N.

2006-01-01

This paper reports on a survey of how Australian undergraduate students perceive the benefits of broad study modes: face-to-face classes, web-based study, and print-based study. Two benefit types were identified through factor analysis: engagement and functionality. Respondents rated face-to-face classes highest on engagement and print-based study…

[Validation of a Japanese version of the Experience in Close Relationship- Relationship Structure].

PubMed

Komura, Kentaro; Murakami, Tatsuya; Toda, Koji

2016-08-01

The purpose of this study was to translate the Experience of Close Relationship-Relationship Structure (ECRRS) and evaluate its validity. In study 1 (N = 982), evidence based internal structure (factor structure, internal consistency, and correlation among sub-scales) and evidence based relations to other variables (depression, reassurance seeking and self-esteem) were confirmed. In study 2 (N = 563), evidence based on internal structure was reconfirmed, and evidence based relations to other variables (IWMS, RQ, and ECR-GO) were confirmed. In study 3 (N = 342), evidence based internal structure (test-retest reliability) was confirmed. Based on these results, we concluded that ECR-RS was valid for measuring adult attachment style.

Quality of reporting web-based and non-web-based survey studies: What authors, reviewers and consumers should consider.

PubMed

Turk, Tarek; Elhady, Mohamed Tamer; Rashed, Sherwet; Abdelkhalek, Mariam; Nasef, Somia Ahmed; Khallaf, Ashraf Mohamed; Mohammed, Abdelrahman Tarek; Attia, Andrew Wassef; Adhikari, Purushottam; Amin, Mohamed Alsabbahi; Hirayama, Kenji; Huy, Nguyen Tien

2018-01-01

Several influential aspects of survey research have been under-investigated and there is a lack of guidance on reporting survey studies, especially web-based projects. In this review, we aim to investigate the reporting practices and quality of both web- and non-web-based survey studies to enhance the quality of reporting medical evidence that is derived from survey studies and to maximize the efficiency of its consumption. Reporting practices and quality of 100 random web- and 100 random non-web-based articles published from 2004 to 2016 were assessed using the SUrvey Reporting GuidelinE (SURGE). The CHERRIES guideline was also used to assess the reporting quality of Web-based studies. Our results revealed a potential gap in the reporting of many necessary checklist items in both web-based and non-web-based survey studies including development, description and testing of the questionnaire, the advertisement and administration of the questionnaire, sample representativeness and response rates, incentives, informed consent, and methods of statistical analysis. Our findings confirm the presence of major discrepancies in reporting results of survey-based studies. This can be attributed to the lack of availability of updated universal checklists for quality of reporting standards. We have summarized our findings in a table that may serve as a roadmap for future guidelines and checklists, which will hopefully include all types and all aspects of survey research.

The Effects of School-Based Condom Availability Programs (CAPs) on Condom Acquisition, Use and Sexual Behavior: A Systematic Review.

PubMed

Wang, Timothy; Lurie, Mark; Govindasamy, Darshini; Mathews, Catherine

2018-01-01

We conducted a systematic review to assess the impact of school-based condom availability programs (CAPs) on condom acquisition, use and sexual behavior. We searched PubMed to identify English-language studies evaluating school-based CAPs that reported process (i.e. number of condoms distributed or used) and sexual behavior measures. We identified nine studies that met our inclusion criteria, with the majority conducted in the United States of America. We judged most studies to have medium risk of bias. Most studies showed that school-based CAPs increased the odds of students obtaining condoms (odds ratios (ORs) for individual studies ranged between 1.81 and 20.28), and reporting condom use (OR 1.36-3.2). Three studies showed that school-based CAPs positively influenced sexual behavior, while no studies reported increase in sexual activity. Findings suggest that school-based CAPs may be an effective strategy for improving condom coverage and promoting positive sexual behaviors.

Consent Procedures and Participation Rates in School-Based Intervention and Prevention Research: Using a Multi-Component, Partnership-Based Approach to Recruit Participants

PubMed Central

Leff, Stephen S.; Franko, Debra L.; Weinstein, Elana; Beakley, Kelly; Power, Thomas J.

2009-01-01

Evaluations of school-based interventions and prevention programs typically require parental consent for students to participate. In school-based efforts, program evaluators may have limited access to parents and considerable effort is required to obtain signed consent. This issue is particularly salient when conducting research in under-resourced, urban schools, where parent involvement in the school setting may be somewhat limited. The aims of this article were to (a) examine the published school-based prevention and intervention literature to assess the state of the field in terms of consent procedures and participation rates; and (b) describe two examples of health promotion studies that used multi-component, partnership-based strategies in urban schools to encourage communication among children, their parents, and researchers. The purpose of the case studies was to generate hypotheses to advance the science related to school-based participant recruitment for research studies. Of nearly 500 studies reviewed, only 11.5% reported both consent procedures and participation rates. Studies using active consent procedures had a mean participation rate of 65.5% (range: 11–100%). This article highlights the need for researchers to report consent procedures and participation rates and describes partnership-based strategies used to enroll students into two urban, school-based health promotion studies. PMID:19834586

Time and learning efficiency in Internet-based learning: a systematic review and meta-analysis.

PubMed

Cook, David A; Levinson, Anthony J; Garside, Sarah

2010-12-01

Authors have claimed that Internet-based instruction promotes greater learning efficiency than non-computer methods. determine, through a systematic synthesis of evidence in health professions education, how Internet-based instruction compares with non-computer instruction in time spent learning, and what features of Internet-based instruction are associated with improved learning efficiency. we searched databases including MEDLINE, CINAHL, EMBASE, and ERIC from 1990 through November 2008. STUDY SELECTION AND DATA ABSTRACTION we included all studies quantifying learning time for Internet-based instruction for health professionals, compared with other instruction. Reviewers worked independently, in duplicate, to abstract information on interventions, outcomes, and study design. we identified 20 eligible studies. Random effects meta-analysis of 8 studies comparing Internet-based with non-Internet instruction (positive numbers indicating Internet longer) revealed pooled effect size (ES) for time -0.10 (p = 0.63). Among comparisons of two Internet-based interventions, providing feedback adds time (ES 0.67, p =0.003, two studies), and greater interactivity generally takes longer (ES 0.25, p = 0.089, five studies). One study demonstrated that adapting to learner prior knowledge saves time without significantly affecting knowledge scores. Other studies revealed that audio narration, video clips, interactive models, and animations increase learning time but also facilitate higher knowledge and/or satisfaction. Across all studies, time correlated positively with knowledge outcomes (r = 0.53, p = 0.021). on average, Internet-based instruction and non-computer instruction require similar time. Instructional strategies to enhance feedback and interactivity typically prolong learning time, but in many cases also enhance learning outcomes. Isolated examples suggest potential for improving efficiency in Internet-based instruction.

The role of feedback in improving the effectiveness of workplace based assessments: a systematic review.

PubMed

Saedon, Habiba; Salleh, Shizalia; Balakrishnan, Arun; Imray, Christopher H E; Saedon, Mahmud

2012-05-02

With recent emphasis placed on workplace based assessment (WBA) as a method of formative performance assessment, there is limited evidence in the current literature regarding the role of feedback in improving the effectiveness of WBA. The aim of this systematic review was to elucidate the impact of feedback on the effectiveness of WBA in postgraduate medical training. Searches were conducted using the following bibliographic databases to identify original published studies related to WBA and the role of feedback: Medline (1950-December 2010), Embase (1980-December 2010) and Journals@Ovid (English language only, 1996-December 2010). Studies which attempted to evaluate the role of feedback in WBA involving postgraduate doctors were included. 15 identified studies met the inclusion criteria and minimum quality threshold. They were heterogeneous in methodological design. 7 studies focused on multi source feedback, 3 studies were based on mini-clinical evaluation exercise, 2 looked at procedural based assessment, one study looked at workplace based assessments in general and 2 studies looked at a combination of 3 to 6 workplace based assessments. 7 studies originated from the United Kingdom. Others were from Canada, the United States and New Zealand. Study populations were doctors in various grades of training from a wide range of specialties including general practice, general medicine, general surgery, dermatology, paediatrics and anaesthetics. All studies were prospective in design, and non-comparative descriptive or observational studies using a variety of methods including questionnaires, one to one interviews and focus groups. The evidence base contains few high quality conclusive studies and more studies are required to provide further evidence for the effect of feedback from workplace based assessment on subsequent performance. There is, however, good evidence that if well implemented, feedback from workplace based assessments, particularly multisource feedback, leads to a perceived positive effect on practice.

Knowledge, skills and attitude of evidence-based medicine among obstetrics and gynaecology trainees: a questionnaire survey.

PubMed

Jeve, Yadava Bapurao; Doshani, Angi; Singhal, Tanu; Konje, Justin

2013-12-01

To determine current evidence-based medicine skills and practice among trainees. Questionnaire study. Electronic survey was sent to all obstetrics and gynaecology trainees in East Midlands South Deanery, and responses collected were anonymous. All obstetrics and gynaecology trainees in East Midland South Deanery. Self-reported attitude, skills and knowledge in various components of evidence-based medicine. 69 trainees were included in the study of which 35 responded. Among all respondents, almost 72% of trainees use non-evidence-based methods to find answers for their clinical questions, whereas only 18% use appropriate evidence-based medicine practice for such queries. Just 35% of trainees have minimum skills of literature searching. Most of the trainees struggle to understand various components of evidence-based medicine. Nearly 80% of trainees do not have formal education or training with regard to evidence-based medicine. This study highlights the inadequacy of evidence-based medicine skills among trainees and urges that evidence-based medicine be incorporated in formal training along with specialty study modules.

The Usability Evaluation of Fakih Method Based on Technology for Students with Hearing Difficulties: The User's Retrospective

ERIC Educational Resources Information Center

Sabdan, Muhammad Sayuti Bin; Alias, Norlidah; Jomhari, Nazean; Jamaludin, Khairul Azhar; DeWitt, Dorothy

2014-01-01

The study is aimed at evaluating the FAKIH method based on technology in teaching al-Quran, based on the user's retrospective. The participants of this study were five students selected based on hearing difficulties. The study employed the user evaluation framework. Teacher's journals were used to determine the frequency and percentage of…

A Review of Case-Based Learning Practices in an Online MBA Program: A Program-Level Case Study

ERIC Educational Resources Information Center

Lee, Seung-hee; Lee, Jieun; Liu, Xiaojing; Bonk, Curt J.; Magjuka, Richard J.

2009-01-01

This study examines how a case-based learning approach was used and facilitated in online business education. Perceptions of students and instructors regarding the practices of case-based learning in online environments are explored in terms of instructional design, facilitation, and technology support. This study finds case-based learning to be a…

How Do Teachers Learn Together? A Study of School-Based Teacher Learning in China from the Perspective of Organisational Learning

ERIC Educational Resources Information Center

Zhang, Xiaolei; Wong, Jocelyn L. N.

2018-01-01

Studies of professional development have examined the influence of school-based approaches on in-service teacher learning and change but have seldom investigated teachers' job-embedded learning processes. This paper explores the dynamic processes of teacher learning in school-based settings. A qualitative comparative case study based on the…

Evaluation of Gene-Based Family-Based Methods to Detect Novel Genes Associated With Familial Late Onset Alzheimer Disease

PubMed Central

Fernández, Maria V.; Budde, John; Del-Aguila, Jorge L.; Ibañez, Laura; Deming, Yuetiva; Harari, Oscar; Norton, Joanne; Morris, John C.; Goate, Alison M.; Cruchaga, Carlos

2018-01-01

Gene-based tests to study the combined effect of rare variants on a particular phenotype have been widely developed for case-control studies, but their evolution and adaptation for family-based studies, especially studies of complex incomplete families, has been slower. In this study, we have performed a practical examination of all the latest gene-based methods available for family-based study designs using both simulated and real datasets. We examined the performance of several collapsing, variance-component, and transmission disequilibrium tests across eight different software packages and 22 models utilizing a cohort of 285 families (N = 1,235) with late-onset Alzheimer disease (LOAD). After a thorough examination of each of these tests, we propose a methodological approach to identify, with high confidence, genes associated with the tested phenotype and we provide recommendations to select the best software and model for family-based gene-based analyses. Additionally, in our dataset, we identified PTK2B, a GWAS candidate gene for sporadic AD, along with six novel genes (CHRD, CLCN2, HDLBP, CPAMD8, NLRP9, and MAS1L) as candidate genes for familial LOAD. PMID:29670507

Evaluation of Gene-Based Family-Based Methods to Detect Novel Genes Associated With Familial Late Onset Alzheimer Disease.

PubMed

Fernández, Maria V; Budde, John; Del-Aguila, Jorge L; Ibañez, Laura; Deming, Yuetiva; Harari, Oscar; Norton, Joanne; Morris, John C; Goate, Alison M; Cruchaga, Carlos

2018-01-01

Gene-based tests to study the combined effect of rare variants on a particular phenotype have been widely developed for case-control studies, but their evolution and adaptation for family-based studies, especially studies of complex incomplete families, has been slower. In this study, we have performed a practical examination of all the latest gene-based methods available for family-based study designs using both simulated and real datasets. We examined the performance of several collapsing, variance-component, and transmission disequilibrium tests across eight different software packages and 22 models utilizing a cohort of 285 families ( N = 1,235) with late-onset Alzheimer disease (LOAD). After a thorough examination of each of these tests, we propose a methodological approach to identify, with high confidence, genes associated with the tested phenotype and we provide recommendations to select the best software and model for family-based gene-based analyses. Additionally, in our dataset, we identified PTK2B , a GWAS candidate gene for sporadic AD, along with six novel genes ( CHRD, CLCN2, HDLBP, CPAMD8, NLRP9 , and MAS1L ) as candidate genes for familial LOAD.

CIRSS vertical data integration, San Bernardino County study phases 1-A, 1-B

NASA Technical Reports Server (NTRS)

Christenson, J.; Michel, R. (Principal Investigator)

1981-01-01

User needs, data types, data automation, and preliminary applications are described for an effort to assemble a single data base for San Bernardino County from data bases which exist at several administrative levels. Each of the data bases used was registered and converted to a grid-based data file at a resolution of 4 acres and used to create a multivariable data base for the entire study area. To this data base were added classified LANDSAT data from 1976 and 1979. The resulting data base thus integrated in a uniform format all of the separately automated data within the study area. Several possible interactions between existing geocoded data bases and LANDSAT data were tested. The use of LANDSAT to update existing data base is to be tested.

Editorial--Avoiding Unethical Helicobacter pylori Clinical Trials: Susceptibility-Based Studies and Probiotics as Adjuvants.

PubMed

Graham, David Y

2015-10-01

As a general rule, any clinical study where the result is already known or when the investigator(s) compares an assigned treatment against another assigned treatment known to be ineffective in the study population (e.g., in a population with known clarithromycin resistance) is unethical. As susceptibility-based therapy will always be superior to empiric therapy in any population with a prevalence of antimicrobial resistance >0%, any trial that randomizes susceptibility-based therapy with empiric therapy would be unethical. The journal Helicobacter welcomes susceptibility or culture-guided studies, studies of new therapies, and studies of adjuvants and probiotics. However, the journal will not accept for review any study we judge to be lacking clinical equipoise or which assign subjects to a treatment known to be ineffective, such as a susceptibility-based clinical trial with an empiric therapy comparator. To assist authors, we provide examples and suggestions regarding trial design for comparative studies, for susceptibility-based studies, and for studies testing adjuvants or probiotics. © 2015 John Wiley & Sons Ltd.

Epidemiological evidence for a health risk from mobile phone base stations.

PubMed

Khurana, Vini G; Hardell, Lennart; Everaert, Joris; Bortkiewicz, Alicja; Carlberg, Michael; Ahonen, Mikko

2010-01-01

Human populations are increasingly exposed to microwave/radiofrequency (RF) emissions from wireless communication technology, including mobile phones and their base stations. By searching PubMed, we identified a total of 10 epidemiological studies that assessed for putative health effects of mobile phone base stations. Seven of these studies explored the association between base station proximity and neurobehavioral effects and three investigated cancer. We found that eight of the 10 studies reported increased prevalence of adverse neurobehavioral symptoms or cancer in populations living at distances < 500 meters from base stations. None of the studies reported exposure above accepted international guidelines, suggesting that current guidelines may be inadequate in protecting the health of human populations. We believe that comprehensive epidemiological studies of long-term mobile phone base station exposure are urgently required to more definitively understand its health impact.

Agent-based real-time signal coordination in congested networks.

DOT National Transportation Integrated Search

2014-01-01

This study is the continuation of a previous NEXTRANS study on agent-based reinforcement : learning methods for signal coordination in congested networks. In the previous study, the : formulation of a real-time agent-based traffic signal control in o...

Evidence-Based Psychosocial Treatments for Child and Adolescent Depression

ERIC Educational Resources Information Center

David-Ferdon, Corinne; Kaslow, Nadine J.

2008-01-01

The evidence-base of psychosocial treatment outcome studies for depressed youth conducted since 1998 is examined. All studies for depressed children meet Nathan and Gorman's (2002) criteria for Type 2 studies whereas the adolescent protocols meet criteria for both Type 1 and Type 2 studies. Based on the Task Force on the Promotion and…

Analysis of Sleep Parameters in Patients with Obstructive Sleep Apnea Studied in a Hospital vs. a Hotel-Based Sleep Center

PubMed Central

Hutchison, Kimberly N.; Song, Yanna; Wang, Lily; Malow, Beth A.

2008-01-01

Background: Polysomnography is associated with changes in sleep architecture called the first-night effect. This effect is believed to result from sleeping in an unusual environment and the technical equipment used to study sleep. Sleep experts hope to decrease this variable by providing a more familiar, comfortable atmosphere for sleep testing through hotel-based sleep centers. In this study, we compared the sleep parameters of patients studied in our hotel-based and hospital-based sleep laboratories. Methods: We retrospectively reviewed polysomnograms completed in our hotel-based and hospital-based sleep laboratories from August 2003 to July 2005. All patients were undergoing evaluation for obstructive sleep apnea. Hospital-based patients were matched for age and apnea-hypopnea index with hotel-based patients. We compared the sleep architecture changes associated with the first-night effect in the two groups. The associated conditions and symptoms listed on the polysomnography referral forms are also compared. Results: No significant differences were detected between the two groups in sleep onset latency, sleep efficiency, REM sleep latency, total amount of slow wave sleep (NREM stages 3 and 4), arousal index, and total stage 1 sleep. Conclusions: This pilot study failed to show a difference in sleep parameters associated with the first-night effect in patients undergoing sleep studies in our hotel and hospital-based sleep laboratories. Future studies need to compare the first-night effect in different sleep disorders, preferably in multi-night recordings. Citation: Hutchison KN; Song Y; Wang L; Malow BA. Analysis of sleep parameters in patients with obstructive sleep apnea studied in a hospital vs. A hotel-based sleep center. J Clin Sleep Med 2008;4(2):119–122. PMID:18468309

A comparison between conventional and LANDSAT based hydrologic modeling: The Four Mile Run case study

NASA Technical Reports Server (NTRS)

Ragan, R. M.; Jackson, T. J.; Fitch, W. N.; Shubinski, R. P.

1976-01-01

Models designed to support the hydrologic studies associated with urban water resources planning require input parameters that are defined in terms of land cover. Estimating the land cover is a difficult and expensive task when drainage areas larger than a few sq. km are involved. Conventional and LANDSAT based methods for estimating the land cover based input parameters required by hydrologic planning models were compared in a case study of the 50.5 sq. km (19.5 sq. mi) Four Mile Run Watershed in Virginia. Results of the study indicate that the LANDSAT based approach is highly cost effective for planning model studies. The conventional approach to define inputs was based on 1:3600 aerial photos, required 110 man-days and a total cost of $14,000. The LANDSAT based approach required 6.9 man-days and cost $2,350. The conventional and LANDSAT based models gave similar results relative to discharges and estimated annual damages expected from no flood control, channelization, and detention storage alternatives.

The temporal stability and predictive validity of affect-based and cognition-based intentions.

PubMed

Keer, Mario; Conner, Mark; Van den Putte, Bas; Neijens, Peter

2014-06-01

Recent research has revealed individual differences in the extent to which people base their intentions on affect and cognition. Two studies are presented that assess whether such differences predict the strength of individuals' intention-behaviour relationships. Participants completed measures of affect, cognition, intention, and behaviour regarding a range of health behaviours. Study 1 (N = 300) found that the strength of the intention-behaviour relationship was significantly related to the extent to which individuals based their intentions on affect, but not to the extent they based them on cognition. Study 2 (N = 387) replicated the findings of the first study. In addition, Study 2 revealed that intention stability mediated the relationship between the degree people based their intentions on affect and the strength of the intention-behaviour relationship. Thus, individuals who base their intentions strongly on affect have more stable intentions, and are therefore more likely to enact them. © 2013 The British Psychological Society.

Interactive Diet and Activity Tracking in AARP (IDATA) Study Data | Division of Cancer Prevention

Cancer.gov

The Interactive Diet and Activity Tracking in AARP (IDATA) Study is a methodologic study of device-based, internet-based, and conventional self-report instrum | Device-based and intensive self-report physical activity and diet data with biomarkers

Using Game-Based Cooperative Learning to Improve Learning Motivation: A Study of Online Game Use in an Operating Systems Course

ERIC Educational Resources Information Center

Jong, Bin-Shyan; Lai, Chien-Hung; Hsia, Yen-Teh; Lin, Tsong-Wuu; Lu, Cheng-Yu

2013-01-01

Many researchers have studied the use of game-based learning. Game-based learning takes many forms, including virtual reality, role playing, and performing tasks. For students to learn specific course content, it is important that the selected game be suited to the course. Thus far, no studies have investigated the use of game-based cooperative…

Morphological configuration of the cranial base among children aged 8 to 12 years.

PubMed

Cossio, Lina; López, Jorge; Rueda, Zulma Vanessa; Botero-Mariaca, Paola

2016-06-14

Cranial base is used as reference structure to determine the skeletal type in cephalometric analysis. The purpose was to assess the cranial base length on lateral cephalic radiographs of children between 8 and 12 and compare these measurements with baseline studies in order to evaluate the relationship between the length and the cranial base angle, articular angle, gonial angle and skeletal type. A Cross-sectional study in 149 children aged 8-12 years, originally from Aburrá Valley, who had lateral cephalic radiographs and consented to participate in this study. The variables studied included: age, sex, sella-nasion, sella-nasion-articular, sella-nasion-basion, articular-gonion-menton, gonion-menton, sella-nasion-point B, sella-nasion-point A y point A-nasion-point B. These variables were digitally measured through i-dixel 2 digital software. One-way ANOVA was used to determine mean values and mean value differences. The values obtained were compared with previous studies. A p value <0.05 was considered significant. Cranial base lengths are smaller in each age and sex group, with differences exceeding 10 mm for measurement, compared both with the study by Riolo (Michigan) and the study carried out in Damasco (Antioquia). No relation was found between the skeletal type and the anterior cranial base length, the sella angle and the cranial base angle. Also, no relation was found between the gonial angle and sella angle or the cranial base angle. The cranial base varies from one population to another. Accordingly, compared to other studies it is shorter for the assessed sample.

Systems engineering studies of lunar base construction

NASA Technical Reports Server (NTRS)

Morgenthaler, George W.

1991-01-01

Many ingenious concepts have been proposed for lunar base construction, but few systematic studies exist which relate time-consistent lunar base construction technologies and the choice of lunar base approach with the long-term SEI objectives - i.e., lunar indigenous base construction and Mars Exploration equipment development. To fill this gap, CSC has taken a two-pronged approach. First, the Center undertook basic geotechnical investigations of lunar soil, fabrication of a scale prototype of a lunar construction crane, a multi-robot construction team laboratory experiment, and a preliminary design of lunar base structures. Second, during Jun. and Jul. 1991 two lunar base construction systems engineering studies were accomplished - a 'near term lunar base' study, and a 'far-term lunar base' study. The goals of these studies were to define the major lunar base construction research problems in consistent technology/construction frameworks, and to define design requirements for construction equipment such as a lunar crane and a regolith mover. The 'near-term lunar base' study examined three different construction concepts for a lunar base comprised of pre-fabricated, pre-tested, Space Station Freedom-type modules, which would be covered with regolith shielding. Concept A used a lunar crane for unloading and transportation; concept B, a winch and cart; and concept C, a walker to move the modules from the landing site to the base site and assemble them. To evaluate the merits of each approach, calculations were made of mass efficiency measure, source mass, reliability, far-term base mass, Mars base mass, and base assembly time. The model thus established was also used to define the requirements for crane speed and regolith mover m(sup 3)/sec rates. A major problem addressed is how to 'mine' the regolith and stack it over the habitats as shielding. To identify when the cost of using indigenous lunar materials to construct the base exceeds the cost of development and delivery of the equipment for processing lunar materials, a study of construction of a candidate sintered regolith 'far term lunar base' was undertaken. A technique was devised for casting slabs of sintered (basaltic) regolith and assembling these into a hemispherical (or geodesic) dome. The major problem occurs with the inner liner. At 14.7 psi and 20 percent oxygen internal atmosphere, the entire structure is in tension, even with the regolith load. Also, another study has indicated that at 14.7 psi major resupply of air will be needed because of leakage, and astronauts may have to engage in extensive pre-breathing and post-breathing for extravehicular activity (EVA) tasks, thus detracting from useful mission work time. An alternative is to operate part of the base at, say, 5 psi and 70 percent oxygen, or to equip the astronauts with hard suits at 8.3 psi or greater. All of these choices directly influence base design and construction techniques.

Virtual reality-based simulators for spine surgery: a systematic review.

PubMed

Pfandler, Michael; Lazarovici, Marc; Stefan, Philipp; Wucherer, Patrick; Weigl, Matthias

2017-09-01

Virtual reality (VR)-based simulators offer numerous benefits and are very useful in assessing and training surgical skills. Virtual reality-based simulators are standard in some surgical subspecialties, but their actual use in spinal surgery remains unclear. Currently, only technical reviews of VR-based simulators are available for spinal surgery. Thus, we performed a systematic review that examined the existing research on VR-based simulators in spinal procedures. We also assessed the quality of current studies evaluating VR-based training in spinal surgery. Moreover, we wanted to provide a guide for future studies evaluating VR-based simulators in this field. This is a systematic review of the current scientific literature regarding VR-based simulation in spinal surgery. Five data sources were systematically searched to identify relevant peer-reviewed articles regarding virtual, mixed, or augmented reality-based simulators in spinal surgery. A qualitative data synthesis was performed with particular attention to evaluation approaches and outcomes. Additionally, all included studies were appraised for their quality using the Medical Education Research Study Quality Instrument (MERSQI) tool. The initial review identified 476 abstracts and 63 full texts were then assessed by two reviewers. Finally, 19 studies that examined simulators for the following procedures were selected: pedicle screw placement, vertebroplasty, posterior cervical laminectomy and foraminotomy, lumbar puncture, facet joint injection, and spinal needle insertion and placement. These studies had a low-to-medium methodological quality with a MERSQI mean score of 11.47 out of 18 (standard deviation=1.81). This review described the current state and applications of VR-based simulator training and assessment approaches in spinal procedures. Limitations, strengths, and future advancements of VR-based simulators for training and assessment in spinal surgery were explored. Higher-quality studies with patient-related outcome measures are needed. To establish further adaptation of VR-based simulators in spinal surgery, future evaluations need to improve the study quality, apply long-term study designs, and examine non-technical skills, as well as multidisciplinary team training. Copyright © 2017 Elsevier Inc. All rights reserved.

Evaluation of cardiovascular risk-lowering health benefits accruing from laboratory-based, community-based and exercise-referral exercise programmes.

PubMed

Webb, R; Thompson, J E S; Ruffino, J-S; Davies, N A; Watkeys, L; Hooper, S; Jones, P M; Walters, G; Clayton, D; Thomas, A W; Morris, K; Llewellyn, D H; Ward, M; Wyatt-Williams, J; McDonnell, B J

2016-01-01

To evaluate the ability of community-based exercise programmes to facilitate public participation in exercise and hence improved cardiovascular health, we assessed the respective impacts of: a continuously monitored exercise programme based within our university (study 1); a Valleys Regional Park-facilitated community-based outdoor exercise programme (study 2); a Wales National Exercise Referral Scheme-delivered exercise-referral programme (study 3). Biomolecular (monocytic PPARγ target gene expression), vascular haemodynamic (central/peripheral blood pressure, arterial stiffness), clinical (insulin sensitivity, blood lipids) and anthropometric (body mass index, waist circumference, heart rate) parameters were investigated using RT-PCR, applanation tonometry, chemical analysis and standard anthropometric techniques. In studies 1-3, 22/28, 32/65 and 11/14 participants adhered to their respective exercise programmes, and underwent significant increases in physical activity levels. Importantly, beneficial effects similar to those seen in our previous studies (eg, modulations in expression of monocytic PPARγ target genes, decreases in blood pressure/arterial stiffness, improvements in blood lipids/insulin sensitivity) were observed (albeit to slightly differing extents) only in participants who adhered to their respective exercise programmes. While study 1 achieved more intense exercise and more pronounced beneficial effects, significant cardiovascular risk-lowering health benefits related to biomolecular markers, blood pressure, arterial stiffness and blood lipids were achieved via community/referral-based delivery modes in studies 2 and 3. Because cardiovascular health benefits were observed in all 3 studies, we conclude that the majority of benefits previously reported in laboratory-based studies can also be achieved in community-based/exercise-referral settings. These findings may be of use in guiding policymakers with regard to introduction and/or continued implementation of community/referral-based exercise programmes.

Editorial - Avoiding unethical Helicobacter pylori clinical trials: Susceptibility-based studies and probiotics as adjuvants

PubMed Central

Graham, David Y.

2016-01-01

As a general rule, any clinical study where the result is already known or when the investigator(s) compares an assigned treatment against another assigned treatment known to be ineffective in the study population (e.g. in a population with known clarithromycin resistance) is unethical. Since susceptibility-based therapy will always be superior to empiric therapy in any population with a prevalence of antimicrobial resistance greater than 0%, any trial that randomizes susceptibility-based therapy with empiric therapy would be unethical. The journal Helicobacter welcomes susceptibility or culture-guided studies, studies of new therapies and of adjuvants and probiotics. However, the Journal will not accept for review any study we judge to be lacking clinical equipoise or which assign subjects to a treatment known to be ineffective, such as a susceptibility-based clinical trial with an empiric therapy comparator. To assist authors we provide examples and suggestion regarding trial design for comparative studies, for susceptibility-based studies, and for studies testing adjuvants or probiotics. PMID:26123529

Analysis of sleep parameters in patients with obstructive sleep apnea studied in a hospital vs. a hotel-based sleep center.

PubMed

Hutchison, Kimberly N; Song, Yanna; Wang, Lily; Malow, Beth A

2008-04-15

Polysomnography is associated with changes in sleep architecture called the first-night effect. This effect is believed to result from sleeping in an unusual environment and the technical equipment used to study sleep. Sleep experts hope to decrease this variable by providing a more familiar, comfortable atmosphere for sleep testing through hotel-based sleep centers. In this study, we compared the sleep parameters of patients studied in our hotel-based and hospital-based sleep laboratories. We retrospectively reviewed polysomnograms completed in our hotel-based and hospital-based sleep laboratories from August 2003 to July 2005. All patients were undergoing evaluation for obstructive sleep apnea. Hospital-based patients were matched for age and apnea-hypopnea index with hotel-based patients. We compared the sleep architecture changes associated with the first-night effect in the two groups. The associated conditions and symptoms listed on the polysomnography referral forms are also compared. No significant differences were detected between the two groups in sleep onset latency, sleep efficiency, REM sleep latency, total amount of slow wave sleep (NREM stages 3 and 4), arousal index, and total stage 1 sleep. This pilot study failed to show a difference in sleep parameters associated with the first-night effect in patients undergoing sleep studies in our hotel and hospital-based sleep laboratories. Future studies need to compare the first-night effect in different sleep disorders, preferably in multi-night recordings.

Water-based exercise training for chronic obstructive pulmonary disease.

PubMed

McNamara, Renae J; McKeough, Zoe J; McKenzie, David K; Alison, Jennifer A

2013-12-18

Land-based exercise training improves exercise capacity and quality of life in people with chronic obstructive pulmonary disease (COPD). Water-based exercise training is an alternative mode of physical exercise training that may appeal to the older population attending pulmonary rehabilitation programmes, those who are unable to complete land-based exercise programmes and people with COPD with comorbid physical and medical conditions. To assess the effects of water-based exercise training in people with COPD. A search of the Cochrane Airways Group Specialised Register of trials, which is derived from systematic searches of bibliographic databases, including the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, CINAHL, AMED and PsycINFO, was conducted (from inception to August 2013). Handsearching was done to identify further qualifying studies from reference lists of relevant studies. Review authors included randomised or quasi-randomised controlled trials in which water-based exercise training of at least four weeks' duration was compared with no exercise training or any other form of exercise training in people with COPD. Swimming was excluded. We used standard methodological procedures expected by The Cochrane Collaboration. Five studies were included with a total of 176 participants (71 people participated in water-based exercise training and 54 in land-based exercise training; 51 completed no exercise training). All studies compared supervised water-based exercise training versus land-based exercise training and/or no exercise training in people with COPD (with average forced expiratory volume in one second (FEV1) %predicted ranging from 39% to 62%). Sample sizes ranged from 11 to 53 participants. The exercise training programmes lasted from four to 12 weeks, and the mean age of participants ranged from 57 to 73 years. A moderate risk of bias was due to lack of reporting of randomisation, allocation and blinding procedures in some studies, as well as small sample sizes.Compared with no exercise, water-based exercise training improved the six-minute walk distance (mean difference (MD) 62 metres; 95% confidence interval (CI) 44 to 80 metres; three studies; 99 participants; moderate quality evidence), the incremental shuttle walk distance (MD 50 metres; 95% CI 20 to 80 metres; one study; 30 participants; high quality evidence) and the endurance shuttle walk distance (MD 371 metres; 95% CI 121 to 621 metres; one study; 30 participants; high quality evidence). Quality of life was also improved after water-based exercise training compared with no exercise (standardised mean difference (SMD) -0.97, 95% CI -0.37 to -1.57; two studies; 49 participants; low quality evidence). Compared with land-based exercise training, water-based exercise training did not significantly change the six-minute walk distance (MD 11 metres; 95% CI -11 to 33 metres; three studies; 62 participants; moderate quality evidence) or the incremental shuttle walk distance (MD 9 metres; 95% CI -15 to 34 metres; two studies; 59 participants; low quality evidence). However, the endurance shuttle walk distance improved following water-based exercise training compared with land-based exercise training (MD 313 metres; 95% CI 232 to 394 metres; two studies; 59 participants; moderate quality evidence). No significant differences were found between water-based exercise training and land-based exercise training for quality of life, as measured by the St George's Respiratory Questionnaire or by three of four domains of the Chronic Respiratory Disease Questionnaire (CRDQ); however, the fatigue domain of the CRDQ showed a statistically significant difference in favour of water-based exercise (MD -3.00; 95% CI -5.26 to -0.74; one study; 30 participants). Only one study reported long-term outcomes after water-based exercise training for quality of life and body composition, and no significant change was observed between baseline results and six-month follow-up results. One minor adverse event was reported for water-based exercise training (based on reporting from two studies; 20 participants). Impact of disease severity could not be examined because data were insufficient. There is limited quality evidence that water-based exercise training is safe and improves exercise capacity and quality of life in people with COPD immediately after training. There is limited quality evidence that water-based exercise training offers advantages over land-based exercise training in improving endurance exercise capacity, but we remain uncertain as to whether it leads to better quality of life. Little evidence exists examining the long-term effect of water-based exercise training.

Systematic Review of Couple-Based HIV Intervention and Prevention Studies: Advantages, Gaps, and Future Directions

PubMed Central

El-Bassel, Nabila

2015-01-01

We conducted a systematic review of couple-based HIV biobehavioral (skills-building, VCT, and adherence) and biomedical (ART, circumcision) prevention and intervention studies designed to reduce sexual-and drug-risk behaviors and HIV transmission and acquisition. Of the 11,162 papers identified in the search, 93 peer-reviewed papers met the inclusion criteria and yielded a total of 33 studies conducted globally. Biobehavioral couple-based prevention and intervention studies have been efficacious in reducing sexual- and drug-risk behaviors, increasing access to HIV testing and care, and improving adherence. Biomedical couple-based studies were found to reduce HIV incidence among HIV-negative sex partners and viral load among HIV-positive partners. Despite much progress, couple-based HIV prevention and intervention studies remain limited; a number of methodological gaps exist and studies focusing on MSM, people who inject drugs, and sex workers are scarce. PMID:24980246

75 FR 27112 - Solicitation of Applications and Notice of Funding Availability for the FRA Railroad System...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-13

... (2) to conduct a Study of the Use of Bio- based Technologies (Lubricants) that can be used in... of the Use of Bio-based Technologies.'' The study will focus on the feasibility of using readily... two grants, ``Locomotive Biofuel Study'' and the ``Study of the Use of Bio-based Technologies...

Does a pre-intervention functional assessment increase intervention effectiveness? A meta-analysis of within-subject interrupted time-series studies.

PubMed

Hurl, Kylee; Wightman, Jade; Haynes, Stephen N; Virues-Ortega, Javier

2016-07-01

This study examined the relative effectiveness of interventions based on a pre-intervention functional behavioral assessment (FBA), compared to interventions not based on a pre-intervention FBA. We examined 19 studies that included a direct comparison between the effects of FBA- and non-FBA-based interventions with the same participants. A random effects meta-analysis of effect sizes indicated that FBA-based interventions were associated with large reductions in problem behaviors when using non-FBA-based interventions as a reference intervention (Effect size=0.85, 95% CI [0.42, 1.27], p<0.001). In addition, non-FBA based interventions had no effect on problem behavior when compared to no intervention (0.06, 95% CI [-0.21, 0.33], p=0.664). Interestingly, both FBA-based and non-FBA-based interventions had significant effects on appropriate behavior relative to no intervention, albeit the overall effect size was much larger for FBA-based interventions (FBA-based: 1.27, 95% CI [0.89, 1.66], p<0.001 vs. non-FBA-based: 0.35, 95% CI [0.14, 0.56], p=0.001). In spite of the evidence in favor of FBA-based interventions, the limited number of comparative studies with high methodological standards underlines the need for further comparisons of FBA-based versus non-FBA-based interventions. Copyright © 2016 Elsevier Ltd. All rights reserved.

The internet as psychological laboratory.

PubMed

Skitka, Linda J; Sargis, Edward G

2006-01-01

This chapter reviews studies published in American Psychological Association (APA) journals from 2003-2004 and additional studies (received in response to listserv requests) that used the Internet to collect data (N=121 total studies). Specific examples of three kinds of Web-based research are reviewed: (a) translational (established methods and research questions are adapted to the Web), (b) phenomenological (behavior on the Web is the focus of study), and (c) novel (methodologically innovations unique to Web-based research). Among other findings, our review indicated that 21% of APA journals published at least one article that reported on Web-based research, most Web-based psychological research uses experimental methods, a surprising number use college student samples, and deception in Web-based research is not uncommon. Strengths and weaknesses of Web-based psychological research in general, and our sample of studies in particular, are reviewed with special attention to possible concerns about sampling and the use of deception.

Community-Based Addiction Treatment Staff Attitudes about the Usefulness of Evidence-Based Addiction Treatment and CBO Organizational Linkages to Research Institutions

ERIC Educational Resources Information Center

Lundgren, Lena; Krull, Ivy; Zerden, Lisa de Saxe; McCarty, Dennis

2011-01-01

This national study of community-based addiction-treatment organizations' (CBOs) implementation of evidence-based practices explored CBO Program Directors' (n = 296) and clinical staff (n = 518) attitudes about the usefulness of science-based addiction treatment. Through multivariable regression modeling, the study identified that identical…

Inquiry-based instruction in secondary science classrooms: A survey of teacher practice

NASA Astrophysics Data System (ADS)

Gejda, Linda Muggeo

The purpose of this quantitative investigation was to describe the extent to which secondary science teachers, who were certified through Connecticut's BEST portfolio assessment process between 1997 and 2004 and had taught secondary science during the past academic year, reported practicing the indicators of inquiry-based instruction in the classroom and the factors that they perceived facilitated, obstructed, or informed that practice. Indicators of inquiry-based instruction were derived from the Biological Sciences Curriculum Study (BSCS) 5E model (Bybee, 1997). The method for data collection was a researcher-developed, self-report, questionnaire entitled "Inquiry-based Instruction in Secondary Science Classrooms: A Survey", which was developed and disseminated using a slightly modified Dillman (2000) approach. Almost all of the study participants reported practicing the 5Es (engage, explore, explain, elaborate, and evaluate) of inquiry-based instruction in their secondary science classrooms. Time, resources, the need to cover material for mandatory assessments, the science topics or concepts being taught, and professional development on inquiry-based instruction were reported to be important considerations in participants' decisions to practice inquiry-based instruction in their science classrooms. A majority of the secondary science teachers participating in this study indicated they had the time, access to resources and the professional development opportunities they needed to practice inquiry-based instruction in their secondary classrooms. Study participants ranked having the time to teach in an inquiry-based fashion and the need to cover material for mandated testing as the biggest obstacles to their practice of inquiry-based instruction in the secondary classroom. Classroom experience and collegial exchange informed the inquiry-based instruction practice of the secondary science teachers who participated in this study. Recommendations for further research, practice, and policy were made based upon the results of this study.

Culture and group-based emotions: could group-based emotions be dialectical?

PubMed

Lu, Minjie; Hamamura, Takeshi; Doosje, Bertjan; Suzuki, Satoko; Takemura, Kosuke

2017-08-01

Group-based emotions are experienced when individuals are engaged in emotion-provoking events that implicate the in-group. This research examines the complexity of group-based emotions, specifically a concurrence of positive and negative emotions, focusing on the role of dialecticism, or a set of folk beliefs prevalent in Asian cultures that views nature and objects as constantly changing, inherently contradictory, and fundamentally interconnected. Study 1 found that dialecticism is positively associated with the complexity of Chinese participants' group-based emotions after reading a scenario depicting a positive intergroup experience. Study 2 found that Chinese participants experienced more complex group-based emotions compared with Dutch participants in an intergroup situation and that this cultural difference was mediated by dialecticism. Study 3 manipulated dialecticism and confirmed its causal effect on complex group-based emotions. These studies also suggested the role of a balanced appraisal of an intergroup situation as a mediating factor.

The Utility of Curriculum-Based Measurement within a Multitiered Framework: Establishing Cut Scores as Predictors of Student Performance on the Alaska Standards-Based Assessment

ERIC Educational Resources Information Center

Legg, David E.

2013-01-01

The purpose of this study was to explore the relationship between student performance on Reading Curriculum-based Measures (R-CBM) and student performance on the Alaska's standards based assessment (SBA) administered to students in Studied School District (SSD) Grade 3 through Grade 5 students in the Studied School District as required by…

Measuring Listening Comprehension Skills of 5th Grade School Students with the Help of Web Based System

ERIC Educational Resources Information Center

Acat, M. Bahaddin; Demiral, Hilmi; Kaya, Mehmet Fatih

2016-01-01

The main purpose of this study is to measure listening comprehension skills of 5th grade school students with the help of web based system. This study was conducted on 5th grade students studying at the primary schools of Eskisehir. The scale used in the process of the study is "Web Based Listening Scale". In the process of the study,…

Counting-On, Trading and Partitioning: Effects of Training and Prior Knowledge on Performance on Base-10 Tasks

ERIC Educational Resources Information Center

Saxton, Matthew; Cakir, Kadir

2006-01-01

Factors affecting performance on base-10 tasks were investigated in a series of four studies with a total of 453 children aged 5-7 years. Training in counting-on was found to enhance child performance on base-10 tasks (Studies 2, 3, and 4), while prior knowledge of counting-on (Study 1), trading (Studies 1 and 3), and partitioning (Studies 1 and…

A SAS-based solution to evaluate study design efficiency of phase I pediatric oncology trials via discrete event simulation.

PubMed

Barrett, Jeffrey S; Jayaraman, Bhuvana; Patel, Dimple; Skolnik, Jeffrey M

2008-06-01

Previous exploration of oncology study design efficiency has focused on Markov processes alone (probability-based events) without consideration for time dependencies. Barriers to study completion include time delays associated with patient accrual, inevaluability (IE), time to dose limiting toxicities (DLT) and administrative and review time. Discrete event simulation (DES) can incorporate probability-based assignment of DLT and IE frequency, correlated with cohort in the case of DLT, with time-based events defined by stochastic relationships. A SAS-based solution to examine study efficiency metrics and evaluate design modifications that would improve study efficiency is presented. Virtual patients are simulated with attributes defined from prior distributions of relevant patient characteristics. Study population datasets are read into SAS macros which select patients and enroll them into a study based on the specific design criteria if the study is open to enrollment. Waiting times, arrival times and time to study events are also sampled from prior distributions; post-processing of study simulations is provided within the decision macros and compared across designs in a separate post-processing algorithm. This solution is examined via comparison of the standard 3+3 decision rule relative to the "rolling 6" design, a newly proposed enrollment strategy for the phase I pediatric oncology setting.

[Research-based thinking of nurses at the end of their basic nursing education].

PubMed

Säämänen, J

1993-01-01

The purpose of this study was to find out the level of research-based thinking that students have internalized during their professional education. The research-based thinking of students has been investigated by analyzing the study reports, they have made at the end of their professional education. "The research-based thinking" is evaluated from the perspective of a research consumer. So, the purpose of this study was not to evaluate the level of studies, but the basis of them. The data consisted of 51 study reports. The data was analyzed by using the content analysis and descriptive statistics. The content analysis was based on the set of general rules for research using deductive methods of approach. The students' problems in research-based thinking fell mainly into the following parts: (1) the students did not use previous studies to build up the frame and the problems of their study, the students used scientific publications limitedly, it seems that the knowledge had the same value to the students regardless of the reference they have taken it from; (2) the students had difficulties in connecting the empirical part of their study with theoretical or conceptual frame. In this study the results indicated, that Finnish nursing students' research-based thinking is not mature in their study-reports at the end of education. It seems important to do further research to be able to analyze the reasons for these findings and how education could be improved in the future.

Getting Personal with Teacher Burnout: A Longitudinal Study on the Development of Burnout Using a Person-Based Approach

ERIC Educational Resources Information Center

Hultell, Daniel; Melin, Bo; Gustavsson, J. Petter

2013-01-01

Studies have suggested that teachers' burnout levels are stable over time. This might be because longitudinal studies on burnout have mainly used a variable-based approach. The purpose of this study was to determine if a person-based approach could provide a more multifaceted perspective to the development of teacher burnout. 816 beginning…

Evaluation of the Professional Development Program on Web Based Content Development

ERIC Educational Resources Information Center

Yurdakul, Bünyamin; Uslu, Öner; Çakar, Esra; Yildiz, Derya G.

2014-01-01

The aim of this study is to evaluate the professional development program on web based content development (WBCD) designed by the Ministry of National Education (MoNE). Based on the theoretical CIPP model by Stufflebeam and Guskey's levels of evaluation, the study was carried out as a case study. The study group consisted of the courses that…

A Case Study of Policies and Procedures to Address Cyberbullying at a Technology-Based Middle School

ERIC Educational Resources Information Center

Tate, Bettina Polite

2017-01-01

This qualitative case study explored the policies and procedures used to effectively address cyberbullying at a technology-based middle school. The purpose of the study was to gain an in-depth understanding of policies and procedures used to address cyberbullying at a technology-based middle school in the southern United States. The study sought…

Effects of Performance-Based Financial Incentives on Work Performance: A Study of Technical-Level Employees in the Private Sector in Sri Lanka

ERIC Educational Resources Information Center

Wickramasinghe, Vathsala; Dabere, Sampath

2012-01-01

The objective of the study is to investigate the effect of performance-based financial incentives on work performance. The study hypothesized that the design features of performance-based financial incentive schemes themselves may influence individuals' work performance. For the study, survey methodology was used and 93 technical-level employees…

Health Education and Symptom Flare Management Using a Video-Based m-Health System for Caring Women with IC/BPS.

PubMed

Lee, Ming-Huei; Wu, Huei-Ching; Tseng, Chien-Ming; Ko, Tsung-Liang; Weng, Tang-Jun; Chen, Yung-Fu

2018-06-10

To assess effectiveness of the video-based m-health system providing videos dictated by physicians for health education and symptom self-management for patients with IC/BPS. An m-health system was designed to provide videos for weekly health education and symptom flare self-management. O'Leary-Sant index and VAS scale as well as SF-36 health survey were administrated to evaluate the disease severity and quality of life (QoL), respectively. A total of 60 IC/BPS patients were recruited and randomly assigned to either control group (30 patients) or study group (30 patients) in sequence depending on their orders to visit our urological clinic. Patients in both control and study groups received regular treatments, while those in the study group received additional video-based intervention. Statistical analyses were conducted to compare the outcomes between baseline and post-intervention for both groups. The outcomes of video-based intervention were also compared with the text-based intervention conducted in our previous study. After video-based intervention, patients in the study group exhibited significant effect manifested in all disease severity and QoL assessments except the VAS pain scale, while no significance was found in the control group. Moreover, the study group exhibited more significant net improvements than the control group in 7 SF-36 constructs, except the mental health. The limitations include short intervention duration (8 weeks) and different study periods between text-based and video-based interventions. Video-based intervention is effective in improving the QoL of IC/BPS patients and outperforms the text-based intervention even in a short period of intervention. Copyright © 2018. Published by Elsevier Inc.

The Relative Effectiveness of Computer-Based and Traditional Resources for Education in Anatomy

ERIC Educational Resources Information Center

Khot, Zaid; Quinlan, Kaitlyn; Norman, Geoffrey R.; Wainman, Bruce

2013-01-01

There is increasing use of computer-based resources to teach anatomy, although no study has compared computer-based learning to traditional. In this study, we examine the effectiveness of three formats of anatomy learning: (1) a virtual reality (VR) computer-based module, (2) a static computer-based module providing Key Views (KV), (3) a plastic…

Designing Web-Based Educative Curriculum Materials for the Social Studies

ERIC Educational Resources Information Center

Callahan, Cory; Saye, John; Brush, Thomas

2013-01-01

This paper reports on a design experiment of web-based curriculum materials explicitly created to help social studies teachers develop their professional teaching knowledge. Web-based social studies curriculum reform efforts, human-centered interface design, and investigations into educative curriculum materials are reviewed, as well as…

College-Based Case Studies in Using PowerPoint Effectively

ERIC Educational Resources Information Center

Inoue-Smith, Yukiko

2016-01-01

This study reexamined PowerPoint's potential to enhance traditional pedagogical practices in higher education. The study addressed (1) the conditions under which PowerPoint meets students' needs in typical lecture-based classrooms, (2) whether professors consider PowerPoint-based lectures more effective than lectures supported by material on…

Review Article: A comparison of flood and earthquake vulnerability assessment indicators

NASA Astrophysics Data System (ADS)

de Ruiter, Marleen C.; Ward, Philip J.; Daniell, James E.; Aerts, Jeroen C. J. H.

2017-07-01

In a cross-disciplinary study, we carried out an extensive literature review to increase understanding of vulnerability indicators used in the disciplines of earthquake- and flood vulnerability assessments. We provide insights into potential improvements in both fields by identifying and comparing quantitative vulnerability indicators grouped into physical and social categories. Next, a selection of index- and curve-based vulnerability models that use these indicators are described, comparing several characteristics such as temporal and spatial aspects. Earthquake vulnerability methods traditionally have a strong focus on object-based physical attributes used in vulnerability curve-based models, while flood vulnerability studies focus more on indicators applied to aggregated land-use classes in curve-based models. In assessing the differences and similarities between indicators used in earthquake and flood vulnerability models, we only include models that separately assess either of the two hazard types. Flood vulnerability studies could be improved using approaches from earthquake studies, such as developing object-based physical vulnerability curve assessments and incorporating time-of-the-day-based building occupation patterns. Likewise, earthquake assessments could learn from flood studies by refining their selection of social vulnerability indicators. Based on the lessons obtained in this study, we recommend future studies for exploring risk assessment methodologies across different hazard types.

Mobile phone base stations-Effects on wellbeing and health.

PubMed

Kundi, Michael; Hutter, Hans-Peter

2009-08-01

Studying effects of mobile phone base station signals on health have been discouraged by authoritative bodies like WHO International EMF Project and COST 281. WHO recommended studies around base stations in 2003 but again stated in 2006 that studies on cancer in relation to base station exposure are of low priority. As a result only few investigations of effects of base station exposure on health and wellbeing exist. Cross-sectional investigations of subjective health as a function of distance or measured field strength, despite differences in methods and robustness of study design, found indications for an effect of exposure that is likely independent of concerns and attributions. Experimental studies applying short-term exposure to base station signals gave various results, but there is weak evidence that UMTS and to a lesser degree GSM signals reduce wellbeing in persons that report to be sensitive to such exposures. Two ecological studies of cancer in the vicinity of base stations report both a strong increase of incidence within a radius of 350 and 400m respectively. Due to the limitations inherent in this design no firm conclusions can be drawn, but the results underline the urgent need for a comprehensive investigation of this issue. Animal and in vitro studies are inconclusive to date. An increased incidence of DMBA induced mammary tumors in rats at a SAR of 1.4W/kg in one experiment could not be replicated in a second trial. Indications of oxidative stress after low-level in vivo exposure of rats could not be supported by in vitro studies of human fibroblasts and glioblastoma cells. From available evidence it is impossible to delineate a threshold below which no effect occurs, however, given the fact that studies reporting low exposure were invariably negative it is suggested that power densities around 0.5-1mW/m(2) must be exceeded in order to observe an effect. The meager data base must be extended in the coming years. The difficulties of investigating long-term effects of base station exposure have been exaggerated, considering that base station and handset exposure have almost nothing in common both needs to be studied independently. It cannot be accepted that studying base stations is postponed until there is firm evidence for mobile phones.

Free-radical sensing by using naphthalimide based mesoporous silica (MCM-41) nanoparticles: A combined fluorescence and cellular imaging study

NASA Astrophysics Data System (ADS)

Jha, Gaurav; Roy, Subhasis; Sahu, Prabhat Kumar; Banerjee, Somnath; Anoop, N.; Rahaman, Abdur; Sarkar, Moloy

2018-01-01

Keeping in mind the advantages of material-based systems over simple molecule-based systems, we have designed and developed three inorganic-organic hybrid systems by anchoring 1,8-naphthalimide derivatives to mesoporous silica nanoparticles for detection of free radicals. Prior to photophysical study, systems are characterized by spectroscopic, microscopic and thermo-gravimetric techniques. Steady state and time-resolved fluorescence studies demonstrate that the hydrazine based system is senstive towards detection of various free radicals. Cellular imaging study reveals cell permeability and toxicity study demonstrates the non-toxic nature of the material. These studies have suggested that present system has the potential to be used in various biological applications.

Nurses' perceptions of evidence-based practice: a quantitative study at a teaching hospital in Iran.

PubMed

Shafiei, Ebrahim; Baratimarnani, Ahmad; Goharinezhad, Salime; Kalhor, Rohollah; Azmal, Mohammad

2014-01-01

Evidence-based practice (EBP) provides nurses a method to use critically appraised and scientifically proven evidence for delivering quality health care and the best decision that leads to quality outcomes. The purpose of this study was to measure the practice, attitude and knowledge/skill of evidence-based practice of nurses in a teaching hospital in Iran. This cross-sectional study was conducted in 2011.The study sample was composed of 195 nurses who were working at the Fatemeh Zahra Hospital affiliated to Bushehr University of Medical Sciences (BPUMS). The survey instrument was a questionnaire based on Upton and Upton study. This tool measures Nurses' perceptions in the three sub-scales of practice, attitude and knowledge/skill of evidence-based practice. Descriptive statistical analysis was used to analyze the data. Pearson correlation coefficients were used to examine the relationship between subscales. The overall mean score of the evidence-based practice in this study was 4.48±1.26 from 7, and the three subscales of practice, attitude and knowledge/skill in evidence-based practice were, 4.58±1.24, 4.57±1.35 and 4.39±1.20, respectively. There was a strong relationship between knowledge and performance subscale (r=0.73,p<0.01). Findings of the study indicate that more training and education are required for evidence-based nursing. Successful implementation of evidence-based nursing depends on organizational plans and empowerment programs in hospitals. Hence, hospital managers should formulate a comprehensive strategy for improving EBP.

A preliminary evaluation of the training effects of a didactic and simulation-based psychological first aid program in students and school counselors in South Korea.

PubMed

Lee, Jong-Sun; You, Sungeun; Choi, Yun-Kyeung; Youn, Hyae-Young; Shin, Hye Sook

2017-01-01

The present study aimed to examine the training effects of a didactic and simulation-based psychological first aid (PFA) program. Based on the competency-based model, the study sought to examine whether the PFA training would enhance knowledge, skills, and attitudes. Study 1 examined the training effects of the PFA program in a sample of undergraduate and graduate students in psychology. Study 2 was conducted with school counselors. In both studies, all participants completed a one-day PFA workshop with a 3-hour didactic lecture and a 3-hour simulation-based practice. Assessments were conducted prior to the didactic lecture and upon completion of the simulation-based practice. In study 1, an examination of pre- and posttest comparisons indicated that the training significantly improved students' PFA knowledge and perceived competence in PFA skill. In study 2, the same PFA training significantly improved school counselors' PFA knowledge, perceived competence in PFA skill, perceived preparedness and confidence to provide psychological assistance for future disasters, but their perceived willingness to participate in psychological assistance did not significantly change after the training. This study provides preliminary evidence supporting the effectiveness of the PFA training program using a combined method of didactic and simulation-based practice for disaster mental health providers in Korea.

A preliminary evaluation of the training effects of a didactic and simulation-based psychological first aid program in students and school counselors in South Korea

PubMed Central

Lee, Jong-Sun; Choi, Yun-Kyeung; Youn, Hyae-young; Shin, Hye Sook

2017-01-01

The present study aimed to examine the training effects of a didactic and simulation-based psychological first aid (PFA) program. Based on the competency-based model, the study sought to examine whether the PFA training would enhance knowledge, skills, and attitudes. Study 1 examined the training effects of the PFA program in a sample of undergraduate and graduate students in psychology. Study 2 was conducted with school counselors. In both studies, all participants completed a one-day PFA workshop with a 3-hour didactic lecture and a 3-hour simulation-based practice. Assessments were conducted prior to the didactic lecture and upon completion of the simulation-based practice. In study 1, an examination of pre- and posttest comparisons indicated that the training significantly improved students’ PFA knowledge and perceived competence in PFA skill. In study 2, the same PFA training significantly improved school counselors’ PFA knowledge, perceived competence in PFA skill, perceived preparedness and confidence to provide psychological assistance for future disasters, but their perceived willingness to participate in psychological assistance did not significantly change after the training. This study provides preliminary evidence supporting the effectiveness of the PFA training program using a combined method of didactic and simulation-based practice for disaster mental health providers in Korea. PMID:28715481

The Impact of a Teaching-Learning Program Based on a Brain-Based Learning on the Achievement of the Female Students of 9th Grade in Chemistry

ERIC Educational Resources Information Center

Shabatat, Kawthar; Al-Tarawneh, Mohammed

2016-01-01

This study aimed at recognizing the impact of teaching-learning program based on a brain-based learning on the achievement of female students of 9th grade in chemistry, to accomplish the goal of this study the researchers designed instruments of: instructional plans, pre achievement and past achievement exams to use them for the study-validity and…

Response efficacy: the key to minimizing rejection and maximizing acceptance of emotion-based anti-speeding messages.

PubMed

Lewis, I M; Watson, B; White, K M

2010-03-01

This study sought to improve understanding of the persuasive process of emotion-based appeals not only in relation to negative, fear-based appeals but also for appeals based upon positive emotions. In particular, the study investigated whether response efficacy, as a cognitive construct, mediated outcome measures of message effectiveness in terms of both acceptance and rejection of negative and positive emotion-based messages. Licensed drivers (N=406) participated via the completion of an on-line survey. Within the survey, participants received either a negative (fear-based) appeal or one of the two possible positive appeals (pride or humor-based). Overall, the study's findings confirmed the importance of emotional and cognitive components of persuasive health messages and identified response efficacy as a key cognitive construct influencing the effectiveness of not only fear-based messages but also positive emotion-based messages. Interestingly, however, the results suggested that response efficacy's influence on message effectiveness may differ for positive and negative emotion-based appeals such that significant indirect (and mediational) effects were found with both acceptance and rejection of the positive appeals yet only with rejection of the fear-based appeal. As such, the study's findings provide an important extension to extant literature and may inform future advertising message design. Copyright 2009 Elsevier Ltd. All rights reserved.

Towards Evidence-Based Practice in Language Intervention for Bilingual Children

ERIC Educational Resources Information Center

Thordardottir, Elin

2010-01-01

Evidence-based practice requires that clinical decisions be based on evidence from rigorously controlled research studies. At this time, very few studies have directly examined the efficacy of clinical intervention methods for bilingual children. Clinical decisions for this population cannot, therefore, be based on the strongest forms of research…

Amniotic fluid embolism mortality rate.

PubMed

Benson, Michael D

2017-11-01

The objective of this study was to determine the mortality rate of amniotic fluid embolism (AFE) using population-based studies and case series. A literature search was conducted using the two key words: 'amniotic fluid embolism (AFE)' AND 'mortality rate'. Thirteen population-based studies were evaluated, as well as 36 case series including at least two patients. The mortality rate from population-based studies varied from 11% to 44%. When nine population-based studies with over 17 000 000 live births were aggregated, the maternal mortality rate was 20.4%. In contrast, the mortality rate of AFE in case series varies from 0% to 100% with numerous rates in between. The AFE mortality rate in population-based studies varied from 11% to 44% with the best available evidence supporting an overall mortality rate of 20.4%. Data from case series should no longer be used as a basis for describing the lethality of AFE. © 2017 Japan Society of Obstetrics and Gynecology.

Qualitative Study of Functional Groups and Antioxidant Properties of Soy-Based Beverages Compared to Cow Milk

PubMed Central

Durazzo, Alessandra; Gabrielli, Paolo; Manzi, Pamela

2015-01-01

Soy-based beverages are a source of high quality proteins and balanced nutrients; they thus represent an alternative to milk in case of allergy to cow milk proteins or intolerance to lactose. In this research, antioxidant properties of soy-based beverages and UHT cow milk were studied. In addition, color parameters, by a quick and non-destructive methodology, were studied in order to verify a possible correlation with antioxidant properties and a qualitative analysis of the major functional groups undertaken by Fourier Transformed Infrared Spectroscopy (FTIR) on Attenuated Total Reflectance (ATR) was carried out. Extractable and hydrolysable polyphenols were studied in soy-based beverages. However, only the extractable fraction was studied in UHT milk, which was characterized by a small amount of polyphenols. All color parameters showed highly significant differences among soy-based beverages and between soy-based beverages and cow milk. FTIR-ATR spectra of soy-based beverages and cow milk showed several differences in the various regions depending on both the specific contribution of molecular groups and different food items. PMID:26783841

Clinical applications of cell-based approaches in alveolar bone augmentation: a systematic review.

PubMed

Shanbhag, Siddharth; Shanbhag, Vivek

2015-01-01

Cell-based approaches, utilizing adult mesenchymal stem cells (MSCs), are reported to overcome the limitations of conventional bone augmentation procedures. The study aims to systematically review the available evidence on the characteristics and clinical effectiveness of cell-based ridge augmentation, socket preservation, and sinus-floor augmentation, compared to current evidence-based methods in human adult patients. MEDLINE, EMBASE, and CENTRAL databases were searched for related literature. Both observational and experimental studies reporting outcomes of "tissue engineered" or "cell-based" augmentation in ≥5 adult patients alone, or in comparison with non-cell-based (conventional) augmentation methods, were eligible for inclusion. Primary outcome was histomorphometric analysis of new bone formation. Effectiveness of cell-based augmentation was evaluated based on outcomes of controlled studies. Twenty-seven eligible studies were identified. Of these, 15 included a control group (8 randomized controlled trials [RCTs]), and were judged to be at a moderate-to-high risk of bias. Most studies reported the combined use of cultured autologous MSCs with an osteoconductive bone substitute (BS) scaffold. Iliac bone marrow and mandibular periosteum were frequently reported sources of MSCs. In vitro culture of MSCs took between 12 days and 1.5 months. A range of autogenous, allogeneic, xenogeneic, and alloplastic scaffolds was identified. Bovine bone mineral scaffold was frequently reported with favorable outcomes, while polylactic-polyglycolic acid copolymer (PLGA) scaffold resulted in graft failure in three studies. The combination of MSCs and BS resulted in outcomes similar to autogenous bone (AB) and BS. Three RCTs and one controlled trial reported significantly greater bone formation in cell-based than conventionally grafted sites after 3 to 8 months. Based on limited controlled evidence at a moderate-to-high risk of bias, cell-based approaches are comparable, if not superior, to current evidence-based bone grafting methods, with a significant advantage of avoiding AB harvesting. Future clinical trials should additionally evaluate patient-based outcomes and the time-/cost-effectiveness of these approaches. © 2013 Wiley Periodicals, Inc.

A Systematic Review of Promising Strategies of Faith-Based Cancer Education and Lifestyle Interventions Among Racial/Ethnic Minority Groups.

PubMed

Hou, Su-I; Cao, Xian

2017-09-13

Church-based interventions have been used to reach racial/ethnic minorities. In order to develop effective programs, we conducted a comprehensive systematic review of faith-based cancer prevention studies (2005~2016) to examine characteristics and promising strategies. Combination terms "church or faith-based or religion," "intervention or program," and "cancer education or lifestyle" were used in searching the five major databases: CINAHL; ERIC; Health Technology Assessments; MEDLINE; and PsycInfo. A total of 20 studies met study criteria. CDC's Community Guide was used to analyze and review group interventions. Analyses were organized by two racial groups: African American (AA) and Latino/Hispanic American groups. Results showed most studies reviewed focused on breast cancer alone or in combination with other cancers. Studies of Latino/Hispanic groups targeted more on uninsured, Medicare, or Medicaid individuals, whereas AA studies generally did not include specific insurance criteria. The sample sizes of the AA studies were generally larger. The majority of these studies reviewed used pre-post, posttest only with control group, or quasi-experience designs. The Health Belief Model was the most commonly used theory in both groups. Community-based participatory research and empowerment/ecological frameworks were also used frequently in the Latino/Hispanic studies. Small media and group education were the top two most popular intervention strategies in both groups. Although one-on-one strategy was used in some Latino studies, neither group used reducing client out-of-pocket costs strategy. Client reminders could also be used more in both groups as well. Current review showed church-based cancer education programs were effective in changing knowledge, but not always screening utilization. Results show faith-based cancer educational interventions are promising. To maximize intervention impact, future studies might consider using stronger study designs, incorporating a variety of proven effective strategies, including those frequently used evidence-based strategies, as well as exploring promising strategies among specific target groups.

Observational studies using propensity score analysis underestimated the effect sizes in critical care medicine.

PubMed

Zhang, Zhongheng; Ni, Hongying; Xu, Xiao

2014-08-01

Propensity score (PS) analysis has been increasingly used in critical care medicine; however, its validation has not been systematically investigated. The present study aimed to compare effect sizes in PS-based observational studies vs. randomized controlled trials (RCTs) (or meta-analysis of RCTs). Critical care observational studies using PS were systematically searched in PubMed from inception to April 2013. Identified PS-based studies were matched to one or more RCTs in terms of population, intervention, comparison, and outcome. The effect sizes of experimental treatments were compared for PS-based studies vs. RCTs (or meta-analysis of RCTs) with sign test. Furthermore, ratio of odds ratio (ROR) was calculated from the interaction term of treatment × study type in a logistic regression model. A ROR < 1 indicates greater benefit for experimental treatment in RCTs compared with PS-based studies. RORs of each comparison were pooled by using meta-analytic approach with random-effects model. A total of 20 PS-based studies were identified and matched to RCTs. Twelve of the 20 comparisons showed greater beneficial effect for experimental treatment in RCTs than that in PS-based studies (sign test P = 0.503). The difference was statistically significant in four comparisons. ROR can be calculated from 13 comparisons, of which four showed significantly greater beneficial effect for experimental treatment in RCTs. The pooled ROR was 0.71 (95% CI: 0.63, 0.79; P = 0.002), suggesting that RCTs (or meta-analysis of RCTs) were more likely to report beneficial effect for the experimental treatment than PS-based studies. The result remained unchanged in sensitivity analysis and meta-regression. In critical care literature, PS-based observational study is likely to report less beneficial effect of experimental treatment compared with RCTs (or meta-analysis of RCTs). Copyright © 2014 Elsevier Inc. All rights reserved.

Evidence based practice beliefs and implementation among nurses: a cross-sectional study

PubMed Central

2014-01-01

Background Having a positive attitude towards evidence-based practice and being able to see the value of evidence-based practice for patients have been reported as important for the implementation of evidence-based practice among nurses. The aim of this study was to map self-reported beliefs towards EBP and EBP implementation among nurses, and to investigate whether there was a positive correlation between EBP beliefs and EBP implementation. Method We carried out a cross-sectional study among 356 nurses at a specialist hospital for the treatment of cancer in Norway. The Norwegian translations of the Evidence-based Practice Belief Scale and the Evidence-based Practice Implementation Scale were used. Results In total, 185 nurses participated in the study (response rate 52%). The results showed that nurses were positive towards evidence-based practice, but only practised it to a small extent. There was a positive correlation (r) between beliefs towards evidence-based practice and implementation of evidence-based practice (r = 0.59, p = 0.001). There was a statistical significant positive, but moderate correlation between all the four subscales of the EBP Beliefs Scale (beliefs related to: 1) knowledge, 2) resources, 3) the value of EBP and 4) difficulty and time) and the EBP Implementation Scale, with the highest correlation observed for beliefs related to knowledge (r = 0.38, p < .0001). Participants who had learned about evidence-based practice had significantly higher scores on the Evidence-based Practice Belief Scale than participants who were unfamiliar with evidence-based practice. Those involved in evidence-based practice working groups also reported significantly higher scores on the Evidence-based Practice Belief Scale than participants not involved in these groups. Conclusion This study shows that nurses have a positive attitude towards evidence-based practice, but practise it to a lesser extent. There was a positive correlation between beliefs about evidence-based practice and implementation of evidence-based practice. Beliefs related to knowledge appear to have the greatest effect on implementation of evidence-based practice. Having knowledge and taking part in evidence-based practice working groups seem important. PMID:24661602

Academic Job Placements in Library and Information Science Field: A Case Study Performed on ALISE Web-Based Postings

ERIC Educational Resources Information Center

Abouserie, Hossam Eldin Mohamed Refaat

2010-01-01

The study investigated and analyzed the state of academic web-based job announcements in Library and Information Science Field. The purpose of study was to get in depth understanding about main characteristics and trends of academic job market in Library and Information science field. The study focused on web-based version announcement as it was…

Intrusion-based reasoning and depression: cross-sectional and prospective relationships.

PubMed

Berle, David; Moulds, Michelle L

2014-01-01

Intrusion-based reasoning refers to the tendency to form interpretations about oneself or a situation based on the occurrence of a negative intrusive autobiographical memory. Intrusion-based reasoning characterises post-traumatic stress disorder, but has not yet been investigated in depression. We report two studies that aimed to investigate this. In Study 1 both high (n = 42) and low (n = 28) dysphoric participants demonstrated intrusion-based reasoning. High-dysphoric individuals engaged in self-referent intrusion-based reasoning to a greater extent than did low-dysphoric participants. In Study 2 there were no significant differences in intrusion-based reasoning between currently depressed (n = 27) and non-depressed (n = 51) participants, and intrusion-based reasoning did not predict depressive symptoms at 6-month follow-up. Interestingly, previously (n = 26) but not currently (n = 27) depressed participants engaged in intrusion-based reasoning to a greater extent than never-depressed participants (n = 25), indicating the possibility that intrusion-based reasoning may serve as a "scar" from previous episodes. The implications of these findings are discussed.

How Affectively-Based and Cognitively-Based Attitudes Drive Intergroup Behaviours: The Moderating Role of Affective-Cognitive Consistency

PubMed Central

Zhou, Jie; Dovidio, John; Wang, Erping

2013-01-01

The moderating role of affective-cognitive consistency in the effects of affectively-based and cognitively-based attitudes on consummatory and instrumental behaviors was explored using two experimental studies in the intergroup context. Study 1 revealed that affectively-based attitudes were better predictors than cognitively-based attitudes regardless of affective-cognitive consistency for consummatory behaviors (e.g., undergraduates’ supportive behaviors toward government officials). Study 2, which investigated task groups’ supportive behaviors toward an immediate supervisory group, found that for these instrumental behaviors cognitively-based attitudes were better predictors than affectively-based attitudes only when affective-cognitive consistency was high. The present research also examined the mechanism by which affective-cognitive consistency moderates the relative roles of affectively-based and cognitively-based attitudes in attitude-behavior consistency. Results indicated that attitude-behavior consistency is eroded primarily because of the weaker relationship of affective or cognitive components to behaviors than to general attitudes. The reciprocal implications of research on attitudes and work on intergroup relations are considered. PMID:24244751

A Cultured Learning Environment: Implementing a Problem- and Service-Based Microbiology Capstone Course to Assess Process- and Skill-Based Learning Objectives

ERIC Educational Resources Information Center

Watson, Rachel M.; Willford, John D.; Pfeifer, Mariel A.

2018-01-01

In this study, a problem-based capstone course was designed to assess the University of Wyoming Microbiology Program's skill-based and process-based student learning objectives. Students partnered with a local farm, a community garden, and a free downtown clinic in order to conceptualize, propose, perform, and present studies addressing problems…

[Physical Activity in the Context of Workplace Health Promotion: A Systematic Review on the Effectiveness of Software-Based in Contrast to Personal-Based Interventions].

PubMed

Rudolph, Sabrina; Göring, Arne; Padrok, Dennis

2018-01-03

Sports and physical activity interventions are attracting considerable attention in the context of workplace health promotion. Due to increasing digitalization, especially software-based interventions that promote physical activity are gaining acceptance in practice. Empirical evidence concerning the efficiency of software-based interventions in the context of workplace health promotion is rather low so far. This paper examines the question in what way software-based interventions are more efficient than personal-based interventions in terms of increasing the level of physical activity. A systematic review according to the specifications of the Cochrane Collaboration was conducted. Inclusion criteria and should-have criteria were defined and by means of the should-have criteria the quality score of the studies was calculated. The software-based and personal-based interventions are presented in 2 tables with the categories author, year, country, sample group, aim of the intervention, methods, outcome and study quality. A total of 25 studies are included in the evaluation (12 personal- and 13 software-based interventions). The quality scores of the studies are heterogeneous and range from 3 to 9 points. 5 personal- and 5 software-based studies achieved an increase of physical activity. Other positive effects on health could be presented in the studies, for example, a reduction in blood pressure or body-mass index. A few studies did not show any improvement in health-related parameters. This paper demonstrates that positive effects can be achieved with both intervention types. Software-based interventions show advantages due to the use of new technologies. Use of desktop or mobile applications facilitate organization, communication and data acquisition with fewer resources needed. A schooled trainer, on the other hand, is able to react to specific and varying needs of the employees. This aspect should be considered as very significant. © Georg Thieme Verlag KG Stuttgart · New York.

[The scientific information that the pharmaceutical industry provides to family doctors].

PubMed

Rivera Casares, F; Richart Rufino, M J; Navas Cutanda, J; Rodríguez Górriz, E; Gómez Moruno, C; Gómez García, B

2005-06-15

To check whether the information in the written publicity that the pharmaceutical industry gives to family doctors really is based on the scientific studies that support it. Cross-sectional study. Health centre on the outskirts of a big city. Over a year, all the scientific studies that laboratory reps gave family doctors along with the advertising for medicines were collected. A total of 63 paired studies and advertising pieces were obtained. 1-3 advertising messages with each supporting study were selected and reviewed in a structured fashion. Then whether or not the messages selected were based on the study was appraised. 44.5% of the advertising messages were not based on the accompanying study; 29.9% clearly were based on the study; and in the rest there was a half-and-half relationship. There was a significant relationship between the evaluation of the advertising messages and the kind of study, masking and the kind of result variable. A high proportion of advertising messages are not based on the study that is reputed to support them. A critique of these studies has to be undertaken before the advertising messages can be looked at.

77 FR 31027 - Submission for OMB Review; Comment Request; Web-Based Assessment of the Clinical Studies Support...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-24

...; Comment Request; Web-Based Assessment of the Clinical Studies Support Center (CSSC) Summary: Under the... current valid OMB control number. Proposed Collection: Title: Web-Based Assessment of the Clinical Studies... Operations and Procedures (MOP); coordinating meeting space and logistics for in-person meetings, Web...

A Pilot Study: Facilitating Cross-Cultural Understanding with Project-Based Collaborative Learning in an Online Environment

ERIC Educational Resources Information Center

Shadiev, Rustam; Hwang, Wu-Yuin; Huang, Yueh-Min

2015-01-01

This study investigated three aspects: how project-based collaborative learning facilitates cross-cultural understanding; how students perceive project-based collaborative learning implementation in a collaborative cyber community (3C) online environment; and what types of communication among students are used. A qualitative case study approach…

Training Teachers to Conduct Trial-Based Functional Analyses

ERIC Educational Resources Information Center

Kunnavatana, S. Shanun; Bloom, Sarah E.; Samaha, Andrew L.; Dayton, Elizabeth

2013-01-01

The trial-based functional analysis (FA) is a promising approach to identification of behavioral function and is especially suited for use in educational settings. Not all studies on trial-based FA have included teachers as therapists, and those studies that have, included minimal information on teacher training. The purpose of this study was to…

CBSS Outreach Project: Computer-Based Study Strategies for Students with Learning Disabilities. Final Report.

ERIC Educational Resources Information Center

Anderson-Inman, Lynne; Ditson, Mary

This final report describes activities and accomplishments of the four-year Computer-Based Study Strategies (CBSS) Outreach Project at the University of Oregon. This project disseminated information about using computer-based study strategies as an intervention for students with learning disabilities and provided teachers in participating outreach…

Project Based Learning in Multi-Grade Class

ERIC Educational Resources Information Center

Ciftci, Sabahattin; Baykan, Ayse Aysun

2013-01-01

The purpose of this study is to evaluate project based learning in multi-grade classes. This study, based on a student-centered learning approach, aims to analyze students' and parents' interpretations. The study was done in a primary village school belonging to the Centre of Batman, already adapting multi-grade classes in their education system,…

77 FR 12598 - Notice Correction; A Multi-Center International Hospital-Based Case-Control Study of Lymphoma in...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-01

... DEPARTMENT OF HEALTH AND HUMAN SERVICES National Institutes of Health Notice Correction; A Multi-Center International Hospital-Based Case-Control Study of Lymphoma in Asia (AsiaLymph) (NCI) The Federal... project titled, ``A multi-center international hospital-based case-control study of lymphoma in Asia (Asia...

The Impact of "Virtualization" on Independent Study Course Completion Rates: The British Columbia Open University Experiment

ERIC Educational Resources Information Center

Giguere, Louis

2009-01-01

In 1997 the British Columbia Open University (BCOU) adopted a virtualization strategy based primarily on twinning off-line independent study distance education courses (textbook-based with study guide and telephone and e-mail tutor support) with alternate online versions (textbook-based with integrated conferencing and communications provided…

Adult Literacy: A Study of Community Based Literacy Programs. Revised and Updated. Volume II: Program Profiles.

ERIC Educational Resources Information Center

Association for Community Based Education, Washington, DC.

This volume accompanies a study of community-based approaches to literacy education for adults. It provides profiles of 26 community-based educational institutions that provided study data. Each profile provides name, address, and telephone number; contact; other project sites; institutional description; mission/purpose; approach; skills…

Action Research Study. A Framework To Help Move Teachers toward an Inquiry-Based Science Teaching Approach.

ERIC Educational Resources Information Center

Staten, Mary E.

This action research study developed a framework for moving teachers toward an inquiry-based approach to teaching science, emphasizing elements, strategies, and supports necessary to encourage and sustain teachers' use of inquiry-based science instruction. The study involved a literature review, participant observation, focus group discussions,…

GEE-based SNP set association test for continuous and discrete traits in family-based association studies.

PubMed

Wang, Xuefeng; Lee, Seunggeun; Zhu, Xiaofeng; Redline, Susan; Lin, Xihong

2013-12-01

Family-based genetic association studies of related individuals provide opportunities to detect genetic variants that complement studies of unrelated individuals. Most statistical methods for family association studies for common variants are single marker based, which test one SNP a time. In this paper, we consider testing the effect of an SNP set, e.g., SNPs in a gene, in family studies, for both continuous and discrete traits. Specifically, we propose a generalized estimating equations (GEEs) based kernel association test, a variance component based testing method, to test for the association between a phenotype and multiple variants in an SNP set jointly using family samples. The proposed approach allows for both continuous and discrete traits, where the correlation among family members is taken into account through the use of an empirical covariance estimator. We derive the theoretical distribution of the proposed statistic under the null and develop analytical methods to calculate the P-values. We also propose an efficient resampling method for correcting for small sample size bias in family studies. The proposed method allows for easily incorporating covariates and SNP-SNP interactions. Simulation studies show that the proposed method properly controls for type I error rates under both random and ascertained sampling schemes in family studies. We demonstrate through simulation studies that our approach has superior performance for association mapping compared to the single marker based minimum P-value GEE test for an SNP-set effect over a range of scenarios. We illustrate the application of the proposed method using data from the Cleveland Family GWAS Study. © 2013 WILEY PERIODICALS, INC.

Lessons learned from a practice-based, multi-site intervention study with nurse participants

PubMed Central

Friese, Christopher R.; Mendelsohn-Victor, Kari; Ginex, Pamela; McMahon, Carol M.; Fauer, Alex J.; McCullagh, Marjorie C.

2016-01-01

Purpose To identify challenges and solutions to the efficient conduct of a multi-site, practice-based randomized controlled trial to improve nurses’ adherence to personal protective equipment use in ambulatory oncology settings. Design The Drug Exposure Feedback and Education for Nurses’ Safety (DEFENS) study is a clustered, randomized, controlled trial. Participating sites are randomized to web-based feedback on hazardous drug exposures in the sites plus tailored messages to address barriers versus a control intervention of a web-based continuing education video. Approach The study principal investigator, the study coordinator, and two site leaders identified challenges to study implementation and potential solutions, plus potential methods to prevent logistical challenges in future studies. Findings Noteworthy challenges included variation in human subjects protection policies, grants and contracts budgeting, infrastructure for nursing-led research, and information technology variation. Successful strategies included scheduled web conferences, site-based study champions, site visits by the principal investigator, and centrally-based document preparation. Strategies to improve efficiency in future studies include early and continued engagement with contract personnel in sites, and proposed changes to the common rule concerning human subjects. The DEFENS study successfully recruited 393 nurses across 12 sites. To date, 369 have completed surveys and 174 nurses have viewed educational materials. Conclusions Multi-site studies of nursing personnel are rare and challenging to existing infrastructure. These barriers can be overcome with strong engagement and planning. Clinical Relevance Leadership engagement, onsite staff support, and continuous communication can facilitate successful recruitment to a workplace-based randomized, controlled behavioral trial. PMID:28098951

Emulsion based cast booster - a priming system

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

Gupta, R.N.; Mishra, A.K.

2005-07-01

This paper explores the potential of emulsion based cast booster to be used as primer to initiate bulk delivered emulsion explosives used in mines. An attempt has been made for comparative study between conventional cast booster and emulsion based cast booster in terms of the initiation process developed and their capability to develop and maintain the stable detonation process in the column explosives. The study has been conducted using a continuous velocity of detonation (VOD) measuring instrument. During this study three blasts have been monitored. In each blast two holes have been selected for study, the first hole being initiatedmore » with conventional cast booster while the other one with emulsion based cast booster. The findings of the study advocates that emulsion based cast booster is capable of efficient priming of bulk delivered column explosive with stable detonation process in the column. Further, the booster had advantages over the conventional PETN/TNT based cast booster. 5 refs., 2 figs., 1 tab., 1 photo.« less

Classroom-Based Physical Activity and Sedentary Behavior Interventions in Adolescents: A Systematic Review and Meta-Analysis.

PubMed

McMichan, Lauren; Gibson, Ann-Marie; Rowe, David A

2018-05-01

It is reported that 81% of adolescents are insufficiently active. Schools play a pivotal role in promoting physical activity (PA) and reducing sedentary behavior (SB). The aim of this systematic review and meta-analysis was to evaluate classroom-based PA and SB interventions in adolescents. A search strategy was developed using the Population Intervention Comparison Outcome Study (PICOS) design framework. Articles were screened using strict inclusion criteria. Study quality was assessed using the Effective Public Health Practice Project quality assessment tool ( http://www.ephpp.ca/tools.html ). Outcome data for preintervention and postintervention were extracted, and effect sizes were calculated using Cohen's d. The strategy yielded 7574 potentially relevant articles. Nine studies were included for review. Study quality was rated as strong for 1 study, moderate for 5 studies, and weak for 3 studies. Five studies were included for meta-analyses, which suggested that the classroom-based interventions had a nonsignificant effect on PA (P = .55, d = 0.05) and a small, nonsignificant effect on SB (P = .16, d = -0.11). Only 9 relevant studies were found, and the effectiveness of the classroom-based PA and SB interventions varied. Based on limited empirical studies, there is not enough evidence to determine the most effective classroom-based methodology to increase PA and SB.

Web Searching: A Process-Oriented Experimental Study of Three Interactive Search Paradigms.

ERIC Educational Resources Information Center

Dennis, Simon; Bruza, Peter; McArthur, Robert

2002-01-01

Compares search effectiveness when using query-based Internet search via the Google search engine, directory-based search via Yahoo, and phrase-based query reformulation-assisted search via the Hyperindex browser by means of a controlled, user-based experimental study of undergraduates at the University of Queensland. Discusses cognitive load,…

A Quantitative Exploration of Preservice Teachers' Intent to Use Computer-based Technology

ERIC Educational Resources Information Center

Kim, Kioh; Jain, Sachin; Westhoff, Guy; Rezabek, Landra

2008-01-01

Based on Bandura's (1977) social learning theory, the purpose of this study is to identify the relationship of preservice teachers' perceptions of faculty modeling of computer-based technology and preservice teachers' intent of using computer-based technology in educational settings. There were 92 participants in this study; they were enrolled in…

The Effectiveness of Using Corpus-Based Materials in Vocabulary Teaching

ERIC Educational Resources Information Center

Paker, Turan; Özcan, Yeliz Ergül

2017-01-01

Our study aimed at finding out the effectiveness of corpus-based vocabulary teaching activities as well as students' attitudes towards concordance-based materials when corpus-based tasks in English vocabulary learning are used. The study was conducted in a preparatory school in a private university. The participants were 28 intermediate level…

Teachers' Experiences with Integrating Play-Based Learning into Standards-Driven Curriculum: A Phenomenological Study

ERIC Educational Resources Information Center

Nugent, Mary Beth Anderson

2017-01-01

The purpose of this phenomenological study was to describe kindergarten teachers' experiences with integrating play-based learning into standards-based academic curriculum in a school district in South Carolina. Play-based learning experiences were defined as instances which allow children to engage in active, social learning experiences in…

Attitudes of Nigerian Secondary School Teachers towards Media-Based Learning.

ERIC Educational Resources Information Center

Ekpo, C. M.

This document presents results of a study assessing the attitudes of secondary school teachers towards media based learning. The study explores knowledge of and exposure to media based learning techniques of a cross section of Nigerian secondary school teachers. Factors that affect the use of media based learning technique are sought. Media based…

Investigating Effects of Problem-Based versus Lecture-Based Learning Environments on Student Motivation

ERIC Educational Resources Information Center

Wijnia, Lisette; Loyens, Sofie M. M.; Derous, Eva

2011-01-01

This study examines the effects of two learning environments (i.e., problem-based learning [PBL] versus lecture-based [LB] environments) on undergraduates' study motivation. Survey results demonstrated that PBL students scored higher on competence but did not differ from LB students on autonomous motivation. Analyses of focus groups further…

Conducting Science Inquiry in Primary Classrooms: Case Studies of Two Preservice Teachers' Inquiry-Based Practices

ERIC Educational Resources Information Center

Leonard, Jacqueline; Boakes, Norma; Moore, Cara M.

2009-01-01

This study examined the impact of an intervention designed to promote inquiry-based instruction among early childhood/elementary preservice teachers in Earth science. Preservice teachers participated in training sessions and community-based internships to deepen Earth science content knowledge and develop inquiry-based practices. Analyses of Earth…