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A SIMULTANEOUS COUPLING AZO-DYE METHOD FOR THE QUANTITATIVE ASSAY OF ESTERASES: BIOCHEMICAL CHARACTERIZATION

Author: Muthanna AA Al-Kaabi مثنى عبد الامير الكعبي
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2012 Volume: 10 Issue: 4 Pages: 306-311
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Background:Enzyme activity is a subject of continuous research. Comparison of data obtained from various quantitative methods needs standardization of techniques in order to verify the results of histochemical and biochemical assays utilized in the study of tissue enzyme activity.Objective:Establishment of a biochemical method for the quantification of enzyme activity in α-naphthyl acetate esterases (ANAE) containing solution using hexazotized pararoseaniline (HP) as a coupling agent.Methods:Wavelength of maximum absorbance (λmax) of coupled HP in solution was analyzed spectrophotometrically based on the simultaneous-coupling method of ANAE demonstration.Results:λmax of the coupled HP was found to be at 425 nm. The relationship between the optical density of the final reaction product (FRP) and the enzyme concentration was linear with the use of azo dye in solution.Conclusion:Data obtained from the biochemical assay of ANAE activity was in agreement with those documented by the histochemical methods in use. Thus, characterization of enzyme activity may be standardized when studying tissue sections and tissue homogenates.Keywords:Esterases, Biochemical assay, Histochemistry, Spectrophotometry


Article
THE PROLIFERATIVE PROFILE OF THE RHOMBENCEPHALICDEMILUNE IN THE DEVELOPING RAT CEREBELLUM: A QUANTITATIVE HISTOCHEMICALSTUDY

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Abstract

BackgroundStudies have shown that the cerebellum develops within the dorsal metencephalon creating a rhombencephalicdemilune (RD) which represents the formation site of the cerebellum granular cells progenitors. These studies used different histological techniques but all have provided qualitative information regarding the biosynthesis and cell mitosis at the RD.ObjectiveQuantifying the proliferative activity of the cellsat the RD during the embryonic period.MethodsSix age groups from day 16 to day 21 albino rat embryos Rattusrattusnorvegicus were investigated with Ag-NOR staining technique to quantify cell proliferation.ResultsThere wasa statistically significant difference (p<0.01) between cellular activity at different age groups with a surge during embryonic day 18.ConclusionsCorrelation with other studies revealed that Ag-NOR staining technique, which reflects protein biosynthesis and nuclear mitotic activity, provided a valuable quantitative measure of cellular proliferation in the developing rat cerebellum.Keywords Rhombencephalicdemilune, Rat, Developing Cerebellum, Ag-NOR, Quantitative, Histochemistry


Article
SEMI-AUTOMATED COMPUTATIONAL METHOD FOR SKELETAL MUSCLE FIBER TYPING WITH LECTINS: CORRELATION WITH MORPHOMETRIC STUDIES

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Abstract

Background :Muscle fiber typing has been an extensive field of study for many years. Though, limited researches applied lectin histochemistry in the clinical diagnosis of muscle disorders; attention was directed mainly towards enzyme histochemistry.Objective:The use of lectins as recognition systems based on specific protein-carbohydrate interactions in correlation with muscle fibers morphometric standards and optical density features to favor the diagnostic procedures of muscle disorders.Methods:Cross-sections of tibialis anterior muscle from 15 adult rats were stained with Con A, PNA, SBA, WGA, SWGA, LFA, UEA-I, and UEA-II lectins. Photographs of stained sections were analyzed with ImageJ 1.44 software for muscle fiber area, perimeter, optical density, and integrated density.Results:There were statistically significant differences between the parameters of muscle fiber types under study (P<0.05) concerning Con A, LFA and UEA-II lectins, but not for the remaining lectins, regarding the optical density and integrated density of muscle fibers.Conclusions:Lectins make accurate recognition of muscle fiber types on fixed paraffin sections when combined with computerized methods to quantify the features seen in muscle biopsies destined for pathological investigations.Key words:Lectins, Muscle fiber typing, Quantitative, Optical density, Morphometry

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