Towards unveiling the nature of short SERPINA1 transcripts: Avoiding the main ORF control to translate alpha1-antitrypsin C-terminal peptides.

Alternative ORFs in-frame with the known genes are challenging to reveal. Yet they may contribute significantly to proteome diversity. Here we focused on the individual expression of the SERPINA1 gene exon 5 leading to direct translation of alpha1-antitrypsin (AAT) C-terminal peptides. The discovery of alternative ways for their production may expand the current understanding of the serpin gene's functioning. We detected short transcripts expressed primarily in hepatocytes. We identified four variants of hepatocyte-specific SERPINA1 short transcripts and individually probed their potential to be translated in living cells. The long mRNA gave the full-length AAT-eGFP fusion, while in case of short transcripts we deduced four active SERPINA1 in-frame alternative ORFs encoding 10, 21, 153 and 169 amino acids AAT C-terminal oligo- and polypeptides. Unlike secretory AAT-eGFP fusion exhibiting classical AAT behavior, truncated AAT-fusions differ by intracellular retention and nuclear enrichment. Immunofluorescence on the endogenous AAT C-terminal epitope showed its accumulation in the cell nucleoli, indicating that short transcripts may be translated in vivo. FANTOM5 CAGE data on SERPINA1 suggest that short transcripts originate from the post-transcriptional cleavage of the spliced mRNA, initiated mainly from the hepatocyte-specific promoter. CONCLUSION: Short SERPINA1 transcripts may represent a source for the direct synthesis of AAT C-terminal peptides with properties uncommon to AAT.

Analysis of neutrophils functional activity in men with mechanical jaundice of various genesis.

Obstructive jaundice (OJ) or blockage of the bile duct code K83.1 (according to ICD 10), occurs in approximately 45-50% of cases of all varieties of jaundice, it can be both non-tumor and tumor genesis. The functional pathway plays a special role in the genesis of complications of breast the activity of neutrophils as key effector cells responsible for the development of the inflammatory process in the breast. Investigation of the metabolic mechanisms of the functioning of neutrophils allows us to identify intracellular targets, when exposed to It was possible to modulate the level of cell reactivity.The study used data from 47 men with obstructive jaundice of non-tumor origin and 45 men with obstructive jaundice of tumor origin (stage I-II of the tumor process). As a control, data from 100 practically healthy men were used. A pronounced change in the kinetics of the chemiluminescent response of neutrophils in men with obstructive jaundice was found, consisting in an increase in the time to reach the maximum intensity, maximum intensity, area under the curve and activation index for both spontaneous and luminol-dependent induced chemiluminescence. The development of the tumor process in this category of patients was accompanied by a decrease in the area parameter under the curve during spontaneous and induced reactions, time to maximum, intensity maximum and activation index during spontaneous chemiluminescence. The data obtained indicate a marked increase in the values of the functional activity of neutrophils in patients with obstructive jaundice of benign origin, as well as a sharp decrease in their values in the presence of a pathological process of malignant origin.

Prolactin Receptor Isoforms as the Basis of Tissue-Specific Action of Prolactin in the Norm and Pathology.

The review describes functional and structural features of different isoforms of prolactin receptor, mechanisms of signaling pathway activation, and molecular messengers involved in the transmission and termination of signal from the prolactin receptor isoforms. Changes in the ratio between prolactin receptor isoforms, key mediators of prolactin signal transduction and termination in various organs and tissues, are analyzed. Special attention is given to the role of molecular mediators and the ratio between the isoforms in normal physiological functions and pathologies. Approaches for therapeutic correction of prolactin signaling impairments are discussed.

Immunomodulatory effects of progesterone and selective ligands of membrane progesterone receptors.

Progesterone (P4) and its analogues regulate various reproductive processes, such as ovulation, implantation, pregnancy maintenance and delivery. In these processes, an important role is played by the immune cells recruited to the female reproductive organs and tissues, where they are exposed to the action of P4. Progestins regulate cellular processes, acting through nuclear steroid receptors (nSRs), membrane P4 receptors (mPRs), and through the sensors. It remains unclear, what type of receptors is used by P4 and its derivatives to exert their effect on the immune cells and how similar their effects are in different types of these cells. We have previously synthesized new progesterone derivatives, among which two selective mPRs ligands, not interacting with nSRs were identified. The objective of this study was to examine the effects of P4 and new selective mPRs ligands on the expression of pro- and anti-inflammatory cytokines in activated human peripheral blood mononuclear cells (PBMCs), THP-1 monocyte cells, and Jurkat T cells. It was demonstrated that the action of P4 and selective ligands was unidirectional, but in different types of the immune cells, their effects were different, and sometimes even opposite. In PBMCs, exposure to these steroids resulted in the increase of mRNA and secreted protein levels of IL-1ß, TNFα, and IL-6 cytokines, as well as in the increase of INFγ mRNA level, decrease of IL-2 mRNA level, increase of TGFß mRNA level, and decrease of IL-4 mRNA and IL-10 secreted protein levels. In monocytes, similarly to PBMCs, expression of IL-1ß and TNFα mRNA was increased, but expression of IL-10 was also increased, and the TGFß expression statistically significantly remained the same. In Jurkat T cells, expression of IL-2 and TNFα mRNA decreased, while expression of IL-10 increased, and expression of TGFß did not change. Thus, progestins act on the immune cells through mPRs and have both pro- and anti-inflammatory effects, depending on the phenotypes of these cells. The data obtained are important for understanding the complexity of the immune system regulation by progestins, which depends on the type of the immune cells and individual characteristics of the immune system.

Agonistic and Antagonistic Effects of Progesterone Derivatives on the Transcriptional Activity of Nuclear Progesterone Receptor B in Yeast Model System.

Identification of progesterone selective agonists and antagonists that act through one of the nuclear progesterone receptor isoforms is of particular importance for the development of tissue-specific drugs in gynecology and anticancer therapy. Fourteen pregna-D'6- and pregna-D'3-pentarane progesterone derivatives with 16α,17α-cycloalkane groups and two progesterone 3-deoxyderivatives were examined for their ability to regulate transcriptional activity of human nuclear progesterone receptor isoform B (nPR-B) expressed in Saccharomyces cerevisiae yeast. Transcriptional activity of nPR-B was measured from the expression of the ß-galactosidase reporter gene with a hormone-responsible element in the promoter. Among the compounds tested, two were full progesterone agonists, four were partial agonists, one compound possessed both agonistic and antagonistic activity, one compound displayed only partial antagonistic activity, and eight compounds did not show any activity. Modifications of the pentarane structure, precisely, introduction of an additional double bound in the A or B rings and/or modification at the 6th position of progesterone, lead to a switch from the complete agonistic activity to partial agonistic or mixed activities. These modifications enable progestins to act as selective modulators of progesterone receptor. Steroids with reduced A-ring and 3-ketogroups lose their ability to regulate PR-B activity. Both 3-deoxycompounds, being selective ligands of progesterone membrane receptors, do not affect PR-B activity.

Changes in the indices of prooxidant and antioxidant systems in blood plasma in men with atrophic gastritis and gastric cancer.

AIM: To study changes in the indices of prooxidant and antioxidant systems in plasma in men with atrophic gastritis and gastric cancer. MATERIALS AND METHODS: The study included 60 healthy men, 42 patients with atrophic gastritis and 50 men, nicardipine patients with gastric cancer stage II according to TNM. All patients underwent serological diagnosis of diffuse atrophic gastritis (definition of pepsinogens and gas- trin-17) and Helicobacter pylori infection. The diagnosis of "atrophic gastritis" was verified by morphological examination of biopsy speci- mens obtained during fibroesophagogastroduodenoscopy. Diagnosis of gastric cancer was carried out in the Krasnoyarsk regional oncologic dispensary on the basis of a comprehensive instrumental and morphological examination. All patients spectrophotometric methods in plasma was determined the content of diene conjugates (DC), malonic dialdehyde (MDA), glutathione-S-transferase (GST), glutathione peroxidase (GPO), superoxide dismutase (SOD) and catalase. RESULTS: The concentration of SOD, GST, GPO and catalase had no significant differences in patients with atrophic gastritis and gastric cancer and prevailed in comparison with healthy persons. Patients with cancer of the stomach content in the blood plasma DK 2.7 times and MDA at 35.2 times higher than healthy individuals, indicating severe oxidative stress in patients with cancer. In patients with atrophic gastritis was ob- served similar but less pronounced pattern. CONCLUSION: The results indicate the presence of oxidative stress in men with atrophic gastritis and gastric cancer.

Role of Na+/K+-ATPase in Natriuretic Effect of Prolactin in a Model of Cholestasis of Pregnancy.

Participation of Na+/K+-ATPase in the natriuretic effect of prolactin in a cholestasis of pregnancy model was investigated. The Na+/K+-ATPase activity in rat kidney medulla, where active sodium reabsorption occurs, decreased in the model of cholestasis of pregnancy and other hyperprolactinemia types compared with intact animals. This effect was not connected with the protein level of α1- and ß-subunits of Na+/K+-ATPase measured by Western blotting in the kidney medulla. Decrease in Na+/K+-ATPase activity in the kidney cortex was not significant, as well as decrease in the quantity of mRNA and proteins of the α1- and ß-subunits of Na+/K+-ATPase. There were no correlations between the Na+/K+-ATPase activity and sodium clearance, although sodium clearance increased significantly in the model of cholestasis of pregnancy and other hyperprolactinemia groups under conditions of stable glomerular filtration rate measured by creatinine clearance. We conclude that the Na+/K+-ATPase is not the only mediator of the natriuretic effect of prolactin in the model of cholestasis of pregnancy.

Selection of Progesterone Derivatives Specific to Membrane Progesterone Receptors.

The search of selective agonists and antagonists of membrane progesterone receptors (mPRs) is a starting point for the study of progesterone signal transduction mechanisms mediated by mPRs, distinct from nuclear receptors. According to preliminary data, the ligand affinity for mPRs differs significantly from that for classical nuclear progesterone receptors (nPRs), which might indicate structural differences in the ligand-binding pocket of these proteins. In the present work, we analyzed the affinity of several progesterone derivatives for mPRs of human pancreatic adenocarcinoma BxPC3 cell line that is characterized by a high level of mPR mRNA expression and by the absence of expression of nPR mRNA. The values were compared with the affinity of these compounds for nPRs. All tested compounds showed almost no affinity for nPRs, whereas their selectivity towards mPRs was different. Derivatives with an additional 19-hydroxyl group and removed 3-keto group had the highest selectivity for mPRs. These results suggest these compounds as the most selective progesterone analogs for studying the mechanisms of progestin action via mPRs.

Human Chorionic Gonadotropin: Unknown about Known.

The last two decade discoveries shift the accent from consideration of human chorionic gonadotripin (hCG) as a hormone, that controls progesterone production by corpus luteum cells, to a powerful paracrine regulator which'in the tandem with its hyperglycozilated analog (hCG-H) induces successful implantation and coordinated dialog between blastocyst and uterus tissues. Ability of hCG to interact with TSH receptor and hCG-H with TGF-beta-RII extend significantly the spectrum of processes controlled by these molecules. Differences between intracellular pathways of signal transduction between hCG and LH mediated by the same receptor (LH/hCG-R) impugn unity of their effector mechanisms previously considered as obvious. Paracine properties-of hCG comprise control of fusing of trophoblasts into syncytiotrophoblasts, angiogenesis, immunity regulation and endometrium predisposition to implantation. Angiogenesis is associated with LH/hCG-R expressed on mural cells of uterine spiral arteries as well as induced secretion of soluble VEGF type by endometrial cells. hCG.regulates ratio between different forms of T-helper cells in maternal organism on the initial gestation stage determining high level of Th2 cells. hCG supports local immunotolerance acting as chemoattractant for T-suppressors (T-Treg) and apoptotic factor for T-lymphocytes. Endometrial susceptibility arises from activation of osteopantin secretion and decline of mucin secretion by epithelial cells. hCG-H acts on the same tissues as hCG as a paracrine agent regulating multiple cascades of cytokines. hCG-H plays the key role in trophoblast invasion into,uterine decidua as a result of gelatinase secretion by these cells.The degree of angiogenic effect of hCG-H is compatiblewith hCG but its signal transduction is mediated by TGF-beta signal transduction pathway that stimulates mural cell proliferation. hCG-H acts as mitogen on NK-cells and is able to activate them and direct to angiogenesis maintenance. In this article the attempt was made to elucidate the most important discoveries about the role of hCG and its hyperglycosylated analog yet accomplished and still upcoming.

Possible Recovery of Manifestation of Prolactin Receptor and Some of Its Target Proteins in the Liver and Kidney Cells of Female Rats after Relief of Cholestasis Complicated and Not Complicated by Hyperprolactinemia.

Immunohistochemistry with semi-quantitative analysis of computer images showed that prolactin receptor and cystic fi brosis transmembrane regulator (CFTR) in cholangiocytes of female rats elevated in cholestasis quickly respond to its relief. The effect of hyperprolactinemia on the extent of return of these proteins to baseline was different. Decompression of the bile duct abolishes the negative effect of hyperprolactinemia on CFTR expression and its positive effect on mrp3 expression in the proximal renal tubules. In renal medulla, mrp2 expression decreased when cholestasis was induced against the background of hyperprolactinemia and increases after its removal. Prolactin receptors and CFTR in cholangiocytes are most susceptible to the decrease in bile duct pressure. Changes in the expression of the studied proteins after cholestasis relief are apparently associated with attenuated toxicity of the products removed by the kidneys, which abolishes the effects of prolactin.

[Predicting of postoperative pain level and morphine consumption by preoperative pressure pain assessment in patients before elective surgery].

UNLABELLED: The aim of this study was to predict a postoperative pain severity and morphine consumption by preoperative pressure pain assessment. DESIGN: 321 patients scheduled for elective surgery (lumbar discectomy, lumbar spinal fusion, hysterectomy, thoracotomy and total hip replacement) in 2009-2013 were enrolled in retrospective study. Pre-operatively, the pain threshold (PTH) and tolerance (PT) in Newton (N) were measured using the pressure algometry. Post-operatively, the pain scores at rest and during movement at 1st postoperative day (POD) using 10 cm VAS were also recorded Patients could get morphine by PCA device in addition to nonopioid analgesia post-operatively. RESULTS: PTH and PT were respectively 34 (24; 45.6) and 74 (54; 95) N, VAS at 1 POD 2 (1; 3.75) at rest and 4 (2,5; 6.25) cm during movement. Pre-operative PT correlated significantly with pain score during movement in patients at 1 POD (R = -0.124, p = 0.026, n = 320). Logit regression analysis found that pain control adequacy during movement at 1 POD could be predicted with PT (ß = 0.011, Std. Err = 0.004, χ2 = 8.536, p = 0.004, OR = 1.011, 95% CI = 1.004-1.018). Morphine consumption by PCA device in patients was 21.25 (7.5; 38) mg. We found a significant correlation between pre-operative PT and post-operative morphine consumption (R = -0.306, p = 0.0006, n = 122). CONCLUSIONS: Post-operative pain severity during movement at 1st postoperative day can be predicted with the pre-operative pain tolerance using the pressure algometry. There is significant moderate negative correlation between pre- operative pain tolerance and post-operative morphine consumption by PCA device in patients at 1st postoperative day.

Expression of Multidrug Resistance Protein 2 (mrp2) in the Liver and Kidney Cells of Female Rats with Modeled Cholestasis of Pregnancy.

Using immunohistochemical method with semiquantitative analysis of images, we showed that mrp2 expression in response to cholestasis decreased in hepatocytes and cholangiocytes, and remained unchanged in the kidney structures. A decrease of mrp2 expression in renal tubules leading to a decrease of metabolic intoxication of the kidney was demonstrated in cholestasis of pregnancy model. In bile ducts cells, negative correlations of mrp2 with previously measured levels of prolactin receptors, CFTR, and mrp3 were revealed. In renal structures and in hepatocytes, no correlations were found between the expression of these proteins. We hypothesize that prolactin produces a direct effect on mrp2 expression in bile ducts cells mediated by prolactin receptors in cholangiocytes. The absence of correlations between mrp2 and the above-mentioned proteins in hepatocytes and renal structures is most likely related to prolactin effects on other systemic regulators.

[Clinical and Immunological Features of Infectious Complications in Patients with Multiple Myeloma].

BACKGROUND: Infectious complications--the leading cause of mortality in patients with multiple myeloma (MM), their appearance is regarded as an adverse prognostic factor in the course of the disease. OBJECTIVE: The aim of our study was to evaluate the clinical and immunological features of infectious complications in patients with G-immunochemical MM to find the most informative indicators in their forecasting. METHODS: A randomized controlled trial was made. All patients were divided into 3 groupsfor comparison: Group 1 (n = 47)--MM patients, G-immunochemical variant with infection, Group 2 (n = 54)--MM patients, G-immunochemical option no infectious complications, and Group 3 (n = 125)--healthy volunteers. Research material was deoxygenated blood taken on admission of a patient to the hematology department before the pathogenetic treatment. Identification of G-variant was carried by immunofixation and electrophoresis. The immune status was assessed by indirect immunofluorescence. The concentration of IgA, M, E and G, and the levels of IL 2, IL 4, IL 8, TNFα, IFNγ in serum was determined by enzyme immunoassay. The activity of neutrophil granulocytes (NG) was studied by chemiluminescent analysis of spontaneous and induced production of reactive oxygen species. Statistical analysis was performed using the software STATISTICA v. 8.0 (USA), RESULTS: We analyzed data from 101 patients with MM and 125 healthy volunteers. The average age of MM patients was 60.53 ± 6.78 years. The group of healthy volunteers was similar in sex and age to groups of patients with MM. In patients with MM in the presence of infectious complications the researchers detected combined secondary development of T and B cell immunodeficiency, changes in non-specific immunity depended on the stage of the disease, unidirectional irregularities in spontaneous and induced chemiluminescence activity NG in II stage disease and multidirectional irregularities in stage III (p = 0.045). Prevalence of the content of proinflammatory cytokines on inflammatory (p < 0.001) and the deviation of the immune response to Th1-type were detected. CONCLUSION: the set of 6 informative indicators (the content of IL 4, IL 2, TNF α, IgG, the absolute number of CD4+ and CD19+ cells) enables the development of prediction method of infectious complications in patients with MM.

The relationship between the pro-oxidant and antioxidant system status of patients with multiple myeloma and the disease stage.

Disturbances of the erythrocyte antioxidant system presented by LPO intensification were detected in patients with multiple myeloma. Plasma concentrations of MDA in these patients were close to normal due to effective work of the nonenzymatic antioxidant system. Activities of antioxidant enzymes in the plasma were reduced, while catalase activity was high, and ceruloplasmin content did not differ from the control, this indicating suppression of the enzymatic component of the antioxidant system. In erythrocytes, the level of reduced glutathione was low, especially at stage III of the disease. Changes in SOD and catalase activities were similar to those in the plasma, while activities of glutathione-dependent enzymes were comparable to those in normal human erythrocytes.

[Chrysotile-asbestos induces cytogenetic effects in the rat's mesothelium in vitro and in vivo].

Under cultivation of rat peritoneal mesothelial cells in vitro in them there are appeared signs of the genomic instability, evidencing their transformation: increasing of the number of both binucleated cells with micronuclei and polynuclear cells and the increase of sizes and polymorphism. Asbestos greatly accelerates this process. Asbestos-induced carcinogenesis in vivo is accompanied in pleural mesothelium in the rats there also revealed with similar signs of genomic instability and cellular transformation.

Manifestation of cystic fibrosis transmembrane regulator (CFTR) in hepatic ductal structures and renal tubules of female rats with experimental cholestasis of pregnancy.

Studies by the immunohistochemical method with semiquantitative analysis of images showed that hyperprolactinemia stimulated CFTR protein manifestation in the bile ducts of female rats, which was clearly expressed in experimental cholestasis of pregnancy. The expression of CFTR in the renal tubules was reduced in hyperprolactinemia under conditions of normal liver function and in cholestasis of pregnancy. Significant positive correlations between CFTR, prolactin receptor, and multiple drug resistance protein 3 were detected in the bile ducts, but not in the renal tubules. Presumably, prolactin has a direct effect on CFTR expression in the bile ducts and indirect effect in the renal tubules. Changes in CFTR protein manifestation in the hepatic ductal structures and renal tubules in experimental pregnancy cholestasis could aggravate the disease.

[The role of cell-mediated immunity in progression of myeloma disease].

The aim of the work was to study the role of cellular and humoral immunity in progression of myeloma disease (MD). The study included 97 patients and 125 age and sex-matched healthy volunteers. The state of cell-mediated and humoral immunity was evaluated at admission, before the pathogenetic treatment, by indirect immunofluorescence technique using anti-CD3, CD4+, CD8+, CD16+, CD19+, HLA-DR+ monoclonal antibodies. Serum IgA, IgM, IgE, IgG were measured by immunoenzymatic assay. The results were analysed using Statistica 7.0 (StatSoft, USA) program. The quantitative values are expressed as the median (Me) and interquartile range (C25-C75). Qualitative variables are presented as absolute values and percentage. The critical statistical significance level was taken as 0.05. The patients with MD developed combined secondary T, B-cell immunodeficiency. The disbalance between CD4+ cells and cytotoxic lymphocytes as well as signs of B-lymphocyte deficit increased with the progress of the disease. The number of NK-cells decreased at all stages of the disease. A rise in the serum level of LgC at all stages compared with control is an identifying sign confirming diagnosis of G-variant of myeloma disease.

[Carcinogenic and mutagenic activity of chrysotile, processed with iron chloride (III)].

This work is devoted to the study of the role of iron ions in the carcinogenic and mutagenic activity of chrysotile. For this aim natural chrysotile was treated with ferric chloride (III), washed, crushed and intratracheally introduced into Wistar rats. When administered to rats intact chrysotile induced mesotheliomas in 27,9 + 4,6% of cases, and chrysotile modified with ferric chloride - in 1,3 +/- 1,3%. Mutagenicity of the same samples was studied in the micronucleus test when administered intraperitoneally to mice Fl (CBA x S57Bl6). Polychromatic erythrocytes in the bone marrow were investigated 24 hours after intraperitoneal administration. The frequency of polychromatic erythrocytes with micronuclei was decreased from 7,4 +/- 0,18 by 1000 due to the action of chrysotile, from 2,8 +/- 0,42 for 1000 after the introduction of a modified sample. It is hypothesized that the ferric chloride modifies the surface of asbestos fibers that reduces the induction of free radicals which are the primary cause of and carcinogenic effects of chrysotile.

Prolactine receptor expression in kidney tissue of female rats with cholestasis: the effect of hyperprolactinemia.

Immunohistochemistry with semiquantitative image analysis showed that prolactin receptor in distal renal tubules of female rats is most sensitive to the negative effects of both cholestasis and hyperprolactinemia. The responses of medullary tubules to cholestasis and hyperprolactinemia were less pronounced: decrease and increase in prolactin receptor expression, respectively. Proximal tubules were characterized by stable levels of prolactin receptor expression insensitive to the effects of obstructive cholestasis and hyperprolactinemia. The cholestasis-induced changes in the intensity of prolactin receptor expression were opposite in kidney and liver cells. It is concluded that different parts of the nephron differ by the presence, type, and direction of regulation of prolactin receptor expression in obstructive cholestasis and hyperprolactinemia.

[Prolactin osmoregulatory function in fishes and its projection on mammals].

Prolactin evolution and key role in fish osmoregulation were reviewed. Comparison of fish and mammalian prolactin was made in respect of its structure, producing tissues, regulation of pituitary secretion. Peculiarities of prolactin receptor structure and prolactin-induced signal cascades, tissue distribution and regulation of prolactin receptor expression were compared in fishes and mammals. Data on mechanisms of prolactin action on ionoconservation in teleost fishes at the level of gills, kidney, intestine, and skin were presented. The facts of prolactin participation in the regulation of water and salt balance in mammals were observed. The existence of fundamentally similar mechanisms of osmoregulatory prolactin action in fishes and mammals was accumed and algorithm of their investigation was suggested.