Assessing the oxidative modification of proteins in inflamed placenta combined with iron deficiency anemia in the pregnant through histochemical method with bromophenol blue based on Mikel Calvo
Y. Karliichuk, , V. Ilika
Department of Pathological Anatomy and Morphology, USA

Aim: To set features of OMB in the cytoplasm of decidua cells in basal plate of the placenta at chorioamnionitis with iron deficiency anemia in pregnant women by means of histochemical methods combined with computer microspectrophotometry.

Introduction: Decidua cells are important cells to the placenta, playing a significant role both in the physiology of pregnancy and during inflammation. The processes of oxidative modification of proteins (OMB) in inflammation are associated with increased levels of oxygen free radicals, which alter the properties of these macromolecules while oxidating amino groups of proteins. Anemic condition is accompanied by intensification of free radical processes in the blood and tissues, and iron deficiency additionally significantly modifies these processes.

Methods: 125 studied placentas, to compare the studied placental physiology of pregnancy and monitoring iron deficiency anemia without inflammation.

A histochemical reaction of bromophenol blue for “acidic” and “basic” proteins by Mikel Calvo was set in histological sections 5μm thick.

Delta Optical Evolution 100 and Olympus SP-550UZ were used to obtain a digital copy of the image. Ratio R/B, which is the ratio between the amino and carboxyl groups in proteins, was determined by “ImageJ”.

Unpaired Student's test calculated arithmetic mean and its error.

Results: When assessing visual histochemical preparations decidua cells are clearly stained, that is suitable for quantitative research, cell boundaries are defined through clear cell membrane coloring and contrasting color around decidua cells fibrinoid. Nuclei and nucleoli were visualized fairly well. “Basic” proteins prevailed in nucleoplasm, while “sour” in the nucleolus.

The decidua cells’ cytoplasm specific color has been mostly granular in nature, and spectral characteristics and optical density of color varied greatly.

Factor R/B at physiological pregnancy (n=20) was – 1.04±0.008 and in iron deficiency anemia (N=21) – 1.06±0.009 P>0.05. In acute chorioamnionitis (n=23) – 1.08±0.009, and combined with iron deficiency anemia (N=21) – 1.09±0.009 P>0.05. Regarding chronic chorioamnionitis (n=20) ratio – 1.24±0.011, and combined with iron deficiency anemia (N=21) – 1.64±0.016 P<0.001.

Conclusion: Conclusion. The intensity of OMB increases only in chronic form of chorioamnionitis in the decidua cells cytoplasm, and combined with iron deficiency anemia significant performance increase has been observed.

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