Histochemical Evaluation of the Processes of Protein Oxidative Modification in the Extravillous Cytotrophoblast of the Utero-Placental Bed during Iron-Deficiency Anemia in Pregnancy

Utero-placental bed is the cumulation of gestationally altered endometrium at the place of ovum attachment to the uterine wall. The key mechanism of this process is the cytotrophoblastic invasion. During iron deficiency anemia, an increase in the specific volume of the extravascular invasive trophoblast is taking place. Concern for the protein oxidative modification in iron deficiency anemia is due to the fact that in conditions of hypoxia, free radical processes in the blood and tissues are enhanced, and iron deficiency is additionally able to modify this problem. The purpose of the study was to establish the histochemical features of the processes of protein oxidative modification in the fractions of extravillous cytotrophoblast of the utero-placental bed depending on the degree of iron deficiency anemia in pregnant women. Material and methods. Quantitative characteristics of protein oxidative modification in the extravillous trophoblast of the utero-placental bed of pregnant women with iron deficiency anemia by means of the histochemical method using reactions with Bromophenol Blue on “acidic” and “basic” proteins according to Mikel Calvo method and computer microdensitometry. We studied 74 biopsies of the utero-placental bed of pregnant women with iron deficiency anemia of I, II and III degrees. The term of gestation was 37-40 weeks. Results and discussion. During physiological pregnancy, the ratio between "acidic" and "basic" proteins in trophoblast cells, even normally, is characterized by a predominance of "acidic" proteins, and evenly in both intravascular and extravascular fractions of cytotrophoblast. Intensification of processes of protein oxidative modification in the cytotrophoblast of the utero-placental bed during iron deficiency anemia of I-II degrees can be assessed as moderate, with an increase in the cells of the endothelium-replacing fraction of cytotrophoblast compared with the interstitial. In conditions of anemia of III degree, a significant predominance of "acidic" proteins in the intravascular cytotrophoblast was noted. Conclusion. During the physiological pregnancy, the intensity of protein oxidative modification was equal in all fractions of the extravillous cytotrophoblast in the utero-placental bed. In the case of gestation with iron deficiency anemia, significant intensification of the protein oxidative modification in the extravillous cytotrophoblast correlated with the severity of anemia. Background iron deficiency anemia significantly affected the processes of protein oxidative modification in the endothelium-replacing cytotrophoblast

Discovery of the Involvement in DNA Oxidative Damage of Human Sperm Nuclear Basic Proteins of Healthy Young Men Living in Polluted Areas

DNA oxidative damage is one of the main concerns being implicated in severe cell alterations, promoting different types of human disorders and diseases. For their characteristics, male gametes are the most sensitive cells to the accumulation of damaged DNA. We have recently reported the relevance of arginine residues in the Cu(II)-induced DNA breakage of sperm H1 histones. In this work, we have extended our previous findings investigating the involvement of human sperm nuclear basic proteins on DNA oxidative damage in healthy males presenting copper and chromium excess in their semen. We found in 84% of those males an altered protamines/histones ratio and a different DNA binding mode even for those presenting a canonical protamines/histones ratio. Furthermore, all the sperm nuclear basic proteins from these samples that resulted were involved in DNA oxidative damage, supporting the idea that these proteins could promote the Fenton reaction in DNA proximity by increasing the availability of these metals near the binding surface of DNA. In conclusion, our study reveals a new and unexpected behavior of human sperm nuclear basic proteins in oxidative DNA damage, providing new insights for understanding the mechanisms related to processes in which oxidative DNA damage is implicated.

Микроскопия с многофотонным возбуждением для идентификации и оперативного контроля компонентов внеклеточного матрикса тканей организма

The process of extraction of the extracellular matrix (ECM) requires operational monitoring since with non-optimal process parameters there is a risk of destruction of the basic proteins of the cell-matrix. Using multiphoton microscopy, the structure of collagen, the integrity of the cell membrane and the content of elastin in the process of isolation of ECM from various tissues of the rat organism (brain, dermis, muscle) were studied. It has been shown that multiphoton microscopy is an effective tool for monitoring the quality of ECM extraction.

Retention Mechanism of Proteins in Hydroxyapatite Chromatography – Multimodal Interaction Based Protein Separations: A Model Study

Background:Retention mechanism of proteins in hydroxyapatite chromatography (HAC) was investigated by linear gradient elution experiments (LGE).Materials and Methods:Several mobile phase (buffer) solution strategies and solutes were evaluated in order to probe the relative contributions of two adsorption sites of hydroxyapatite (HA) particles, C-site due to Ca (metal affinity) and P-site due to PO4 (cation-exchange). When P-site was blocked, two basic proteins, lysozyme (Lys) and ribonuclease A(RNase), were not retained whereas cytochrome C(Cyt C) and lactoferrin (LF) were retained and also retention of acidic proteins became stronger as the repulsion due to P-site was eliminated. The number of the binding site B values determined from LGE also increased, which also showed reduction of repulsion forces.Conclusion:The selectivity (retention) of four basic proteins (RNase, Lys, Cyt C, LF) in HAC was different from that in ion-exchange chromatography. Moreover, it was possible to tune the selectivity by using NaCl gradient.