The effect of prostaglandin induced labor and selected factors on the occurrence and extent of birth-canal injuries.

OBJECTIVES: To determine the effect of labor induction using prostaglandin, gestation period and delivery type on the cause and extent of birth-canal injuries. METHODS: A retrospective study based on the records of II. GPK UNB Ruzinov. The total set (n = 1377) consisted of patients who went into labor in second half of the year 2014 at this clinic. The research used categorical data divided into groups of factors (induction, week of delivery, type of delivery) and birth injuries. Using the chi-square test and Fisher's test we evaluated individual statistical dependences. The results with the highest level of significance are expressed through contingency tests (Phi coefficient, contingency coefficient, Cramer's V). Finally, because of the highest level of significance we have done a cohort study to express the incidence of relationship between non-operative delivery per vaginam and 1st degree ruptura perieni. RESULTS: Labor induction with prostaglandins has a positive impact on the cause and extent of birth-canal injuries, namely the increased risk of vaginal walls tearing and uterine ruptures. We confirmed that the induction of labor using PG carries a demonstrable risk of pregnancy termination by cesarean section (p = 6.17x10-9). Birth-canal injuries are also affected by the type of delivery. Non-operative vaginal delivery is a significant risk factor for first-degree perineal tear (RR = 25.52 95% CI 10.58, 61.60, OR = 33.06 95% CI 13.51, 80.90). Significant risk factors for third-grade perineal tear during vaginal delivery are forceps (p = 0.005534) / vacuum extractor (p = 0.03554). CONCLUSION: In a group of 1377 patients we have demonstrated that the labor induction with prostaglandins, gestation period and type of delivery significantly influence the occurrence and extent of birth-canal injuries (Tab. 3, Ref. 9).

Postpartum corticosteroids in HELLP syndrome - standard to prompt recovery.

OBJECTIVES: To perform a complex review of HELLP syndrome and its treatment from up to date scientific literature. Study of the interresting treatement regime of the patient. METHODS: Complex analysis of the syndrome, systematic search of medical scientific databases and Slovak Medical Library. Analysis of the life threatening state of the patient, retrospective analysis of the diagnostics, treatment, acute management, complications and clinical results. RESULTS: According to up to date literature and our good clinical experiences we can encourage the use of high-dose corticosteroid therapy in HELLP syndrome. CONCLUSION: The most recent studies definitely recommend the high dose corticosteroid treatment in recovery management by the HELLP syndrome. We have used the high-dose corticosteroid regime as a recovery management for the patient with postpartum HELLP with very good clinical and laboratory response followed by prompt recovery of the patient and without other complications. We want to support and empasize the indication of the Dexamethasone regiment by HELLP, becouse the clinical experiences with this treatment are not well-known and usually the corticosteroids are not given to the patients with HELLP, or the treatment is "daemonized". Our clinical experience with this treatment was successful and the patient definitely profited of it. We support the opinion, that the benefit of described regiment highly exceedes the possible adverse effects of the therapy (Fig. 8, Ref. 61).

Endothelial Caveolin-1 regulates the radiation response of epithelial prostate tumors.

The membrane protein caveolin-1 (Cav1) recently emerged as a novel oncogene involved in prostate cancer progression with opposed regulation in epithelial tumor cells and the tumor stroma. Here we examined the role of stromal Cav1 for growth and radiation response of MPR31-4 prostate cancer xenograft tumors using Cav1-deficient C57Bl/6 mice. Syngeneic MPR31-4 tumors grew faster when implanted into Cav1-deficient mice. Increased tumor growth on Cav1-deficient mice was linked to decreased integration of smooth muscle cells into the wall of newly formed blood vessels and thus with a less stabilized vessel phenotype compared with tumors from Cav1 wild-type animals. However, tumor growth delay of MPR31-4 tumors grown on Cav1 knockout mice to a single high-dose irradiation with 20 Gray was more pronounced compared with tumors grown on wild-type mice. Increased radiation-induced tumor growth delay in Cav1-deficient mice was associated with an increased endothelial cell apoptosis. In vitro studies using cultured endothelial cells (ECs) confirmed that the loss of Cav1 expression increases sensitivity of ECs to radiation-induced apoptosis and reduces their clonogenic survival after irradiation. Immunohistochemical analysis of human tissue specimen further revealed that although Cav1 expression is mostly reduced in the tumor stroma of advanced and metastatic prostate cancer, the vascular compartment still expresses high levels of Cav1. In conclusion, the radiation response of MPR31-4 prostate tumors is critically regulated by Cav1 expression in the tumor vasculature. Thus, Cav1 might be a promising therapeutic target for combinatorial therapies to counteract radiation resistance of prostate cancer at the level of the tumor vasculature.

Loss of caveolae, vascular dysfunction, and pulmonary defects in caveolin-1 gene-disrupted mice.

Caveolae are plasma membrane invaginations that may play an important role in numerous cellular processes including transport, signaling, and tumor suppression. By targeted disruption of caveolin-1, the main protein component of caveolae, we generated mice that lacked caveolae. The absence of this organelle impaired nitric oxide and calcium signaling in the cardiovascular system, causing aberrations in endothelium-dependent relaxation, contractility, and maintenance of myogenic tone. In addition, the lungs of knockout animals displayed thickening of alveolar septa caused by uncontrolled endothelial cell proliferation and fibrosis, resulting in severe physical limitations in caveolin-1-disrupted mice. Thus, caveolin-1 and caveolae play a fundamental role in organizing multiple signaling pathways in the cell.

Endothelial-cell permeability and protein kinase C in pre-eclampsia.

BACKGROUND: Oedema and vascular leakage play a part in the pathogenesis of pre-eclampsia. We tested the hypothesis that serum from pre-eclamptic patients increases endothelial-cell permeability and examined possible signal-transduction pathways. METHODS: We studied eight patients with pre-eclampsia, eight normotensive pregnant women, eight non-pregnant women, five pregnant patients with pre-existing hypertension, and four hypertensive non-pregnant women. Cultured human umbilical-vein endothelial-cell monolayers were used and permeability was measured by albumin flux. The part played by protein kinase C (PKC) signalling was examined by down-regulation with phorbol ester and with the inhibitors Goe 6976 and staurosporine. PKC isoforms were assessed by western blot and confocal microscopy. Antisense oligodesoxynucleotides (ODN) were used to test for specific PKC isoforms. FINDINGS: Serum from pre-eclamptic women increased endothelial permeability significantly (by 100%, p<0.01). The change in permeability decreased rapidly after delivery. Serum from normotensive pregnant women and non-pregnant women had no effect. Permeability was not influenced by serum from patients with essential hypertension or pregnant patients with pre-existing hypertension. Serum from pre-eclamptic patients induced a translocation of PKC isoforms alpha and epsilon within the cells. Goe 6976 and staurosporine (10(-8) mol/L) inhibited the increase in permeability induced by serum from pre-eclamptic patients. Down-regulation of PKC alpha and, to a lesser extent, PKC epsilon by antisense ODN also inhibited the pre-eclampsia-induced permeability increase. INTERPRETATION: Serum from pre-eclamptic patients contains a factor or factors that increase endothelial-cell permeability. The effect of pre-eclamptic serum may be mediated by PKC alpha and epsilon.

Effects of AT1 receptor blockade on blood pressure and the renin-angiotensin system in spontaneously hypertensive rats of the stroke prone strain.

The aim of the study was to assess the effects of chronic angiotensin I receptor blockade on blood pressure, the renin-angiotensin system in plasma and kidney and the extent of renal damage in spontaneously hypertensive rats of the stroke prone strain (SHRsp). Four months old male SHRsp rats were orally treated with a high (10 mg/kg b.w. per day) or a low dose (1 mg/kg b.w. per day) of the AT1 receptor antagonist Telmisartan and compared to Losartan- (20 mg/kg b.w. per day), Captopril-treated (50 mg/kg b.w. per day) or untreated control groups for 38 days. Despite a similar extent of blood pressure reduction in all groups (except low dose Telmisartan), high dose Telmisartan but not Losartan or Captopril significantly reduced left ventricular weight by 24% compared to controls (p<0.05). Renal damage as assessed by urinary albumin or glomerulosclerosis index was significantly reduced in all treatment groups (p<0.02). Plasma renin concentration was significantly elevated (p<0.02) and plasma angiotensinogen significantly lowered (p<0.05) in all pharmacologically treated group compared to controls. In the kidney, renin-mRNA as well as AT1 receptor gene expression were elevated in all treatment groups, but no significant changes were found for renal angiotensinogen-mRNA. Chronic oral treatment of genetically hypertensive rats by the AT1 receptor antagonist Telmisartan reveals a blood pressure lowering and reno-protective effect of this drug comparable to other AT1 receptor antagonists or converting enzyme inhibitors, and demonstrates a marked reduction of cardiac hypertrophy by Telmisartan in this model.

From totipotent embryonic stem cells to spontaneously contracting smooth muscle cells: a retinoic acid and db-cAMP in vitro differentiation model.

Vascular smooth muscle cell (VSMC) differentiation is important in understanding vascular disease; however, no in vitro model is available. Totipotent mouse embryonic stem (ES) cells were used to establish such a model. To test whether the ES cell-derived smooth muscle cells expressed VSMC-specific properties, the differentiated cells were characterized by 1) morphological analysis, 2) gene expression, 3) immunostaining for VSMC-specific proteins, 4) expression of characteristic VSMC ion channels, and 5) formation of [Ca2+]i transients in response to VSMC-specific agonists. Treatment of embryonic stem cell-derived embryoid bodies with retinoic acid and dibutyryl-cyclic adenosine monophosphate (db-cAMP) induced differentiation of spontaneously contracting cell clusters in 67% of embryoid bodies compared with 10% of untreated controls. The highest differentiation rate was observed when retinoic acid and db-cAMP were applied to the embryoid bodies between days 7 and 11 in combination with frequent changes of culture medium. Other protocols with retinoic acid and db-cAMP, as well as single or combined treatment with VEGF, ECGF, bFGF, aFGF, fibronectin, matrigel, or hypoxia did not influence the differentiation rate. Single-cell RT-PCR and sequencing of the PCR products identified myosin heavy chain (MHC) splice variants distinguishing between gut and VSMC isoforms. RT-PCR with VSMC-specific MHC primers and immunostaining confirmed the presence of VSMC transcripts and MHC protein. Furthermore, VSMC expressing MHC had typical ion channels and responded to specific agonists with an increased [Ca2+]i. Here we present a retinoic acid + db-cAMP-inducible embryonic stem cell model of in vitro vasculogenesis. ES cell-derived cells expressing VSMC-specific MHC and functional VSMC properties may be a suitable system to study mechanisms of VSMC differentiation.

Mammalian homologues of C. elegans PAR-1 are asymmetrically localized in epithelial cells and may influence their polarity.

The establishment of polarity in the embryo is fundamental for the correct development of an organism [1]. The first cleavage of the Caenorhabditis elegans embryo is asymmetric with certain cytoplasmic components being distributed unequally between the daughter cells [2-4]. Using a genetic screen, Kemphues and co-workers have identified six par genes (partition-defective) [5,6], which are involved in the process of asymmetric division. One of these genes encodes a highly conserved protein, PAR-1, which is a serine/threonine kinase that localizes asymmetrically to the posterior part of the zygote and to those blastocysts that give rise to the germ line [7-9]. We reasoned that the mammalian homologue of PAR-1 (mPAR-1) might be involved in the process of polarization of epithelial cells, which consist of apical and basolateral membrane domains. We found that mPAR-1 was expressed in a wide variety of epithelial tissues and cell lines and was associated with the cellular cortex. In polarized epithelial cells, mPAR-1 was asymmetrically localized to the lateral domain. A fusion protein lacking the kinase domain had the same localization as the full-length protein but its prolonged expression acted in a dominant-negative fashion: lateral adhesion of the transfected cells to neighbouring cells was diminished, resulting in the former cells being 'squeezed out' from the monolayer. Moreover, the polarity of these cells was disturbed resulting in mislocalization of E-cadherin. Thus, in the C. elegans embryo and in epithelial cells, polarity appears to be governed by similar mechanisms.

Endothelial adhesion molecules and leukocyte integrins in preeclamptic patients.

Endothelial cell activation is important in the pathogenesis of preeclampsia; however, the nature of the activation is unknown. We investigated 22 patients with preeclampsia. 29 normotensive pregnancies, and 18 nonpregnant women to test the hypothesis that serum from preeclamptic patients induces expression of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1) and stimulates intracellular free calcium concentrations [Ca2+]i in cultured endothelial cells. We then asked whether the corresponding integrin adhesive counter receptors lymphocyte function-associated antigen-1 (CD11a/CD18), macrophage-1 antigen (CD11b/CD18), p150,95 (CD11c/CD18), and very late activation antigen-4 (CD49/CD29) are increased in patients with preeclampsia. In the pregnant women, the measurements were conducted both before and after delivery. Integrin expression was measured by fluorescent antibody cell sorting analysis using monoclonal antibodies. ICAM-1 and VCAM-1 were analyzed on endothelial cells by enzyme-linked immunosorbent assay. [Ca2+]i was measured with fura 2. Serum from preeclamptic patients increased endothelial cell ICAM-1 expression but not VCAM-1 expression. Preeclamptic patients' serum also increased [Ca2+]i in endothelial cells compared with serum from normal nonpregnant or normal pregnant women. Endothelial cell [Ca2+]i concentrations were correlated with the ICAM-1 expression in preeclamptic patients (r = .80, P < .001) before but not after delivery. Expression of the integrin counter receptors on leukocytes was similarly increased in preclampsia and normal pregnancy compared with the nonpregnant state. The expression decreased significantly after delivery in both groups. Our results demonstrate that serum from preeclamptic women induces increased ICAM-1 surface expression on endothelial cells, while the expression of the integrin counterreceptors was not different. The effect on endothelial cells may be related to an increase in [Ca2+]i. The effect on cultured endothelial cells and the rapid decrease after delivery suggests the presence of a circulating serum factor which increases endothelial cell [Ca2+]i and enhances adhesion molecule expression.

The role of hyperglycemia and hyperinsulinemia in the pathogenesis of diabetic angiopathy.

The small and large vessel disease associated with diabetes mellitus is responsible for its morbidity and mortality. Although much of the pathogenesis remains to be clarified, the role of hyperinsulinemia and hyperglycemia per se in the progression of vascular disease is beginning to emerge. Hyperinsulinemia increases the release of very low density lipoprotein (VLDL) and may also be responsible for the low HDL cholesterol levels in patients with diabetes. Hyperinsulinemia also contributes to increased blood pressure, which independently promotes vascular disease. High glucose concentrations have direct influence on intracellular signal transduction, including effects on sorbitol pathway and associated changes of pyridine nucleotides, the de novo synthesis of diacylglycerol with subsequent stimulation of protein kinase C, and possibly changes in the cellular generation of myoinositol. Hyperglycemia also exerts long-lasting changes in cellular function, which result from non-enzymatic glycosylation of matrix and membrane proteins with subsequent binding of these proteins to specific receptors. These receptors are termed the advanced glycosylation end-products (AGE) receptors. Their activation leads to an increased release of cytokines and growth factors including PDGF, interleukins, TNF-alpha, and TGF-beta, all of which may act concomitantly in the disease process.

PCR analysis of human renal biopsies--renin gene regulation in glomerulonephritis.

Animal studies show that the renin-angiotensin system contributes to hypertension, glomerulosclerosis and progressive chronic renal failure in renal disease. Direct data on the activity of the renal renin-angiotensin system (RAS) in human renal disease are scarce, however. The small amount of tissue in renal biopsies and insufficient sensitivity of analytical methods have precluded reliable measurements of the tissue components of the RAS in the past. Due to its highly sensitivity the quantitative polymerase chain reaction (QPCR) assay today allows the quantitation of gene transcription products in small tissue samples, for example, renal biopsies. The clinical applicability of the PCR has raised the interest in this methodology. We adopted quantitative PCR to study expression and regulation of the renin gene in patients with different forms of glomerulonephritis. For PCR a deletion mutant of the renin gene was used as an internal standard exhibiting the same primer binding sites as the human gene. The number of glomeruli per biopsy sample was counted by microscopic transillumination immediately after biopsy and was used as a reference base. Renin mRNA was expressed as fg per glomerulus. Compared to nonaffected tissue of tumor nephrectomy samples, renin gene expression was significantly lower in glomerulonephritic patients without converting enzyme inhibitor (CEI) treatment, that is, 63 +/- 20 (6) versus 250 +/- 50 (7) (P < 0.02), although plasma renin concentration was in the normal range. Significantly higher renin mRNA expression was found in patients with glomerulonephritis treated with CEI, that is, 210 +/- 50 (8) versus 63 +/- 20 (6) in patients not treated.(ABSTRACT TRUNCATED AT 250 WORDS)

Water immersion-induced endocrine alterations in women with EPH gestosis.

In 12 healthy pregnant women, 14 women with mild or moderate late pregnancy gestosis (EPH) and in 12 non-pregnant women, the influence of head out water immersion (WI) on mean blood pressure (MAP), the renin-aldosterone system, vasopressin (AVP) and atrial natriuretic hormone (ANF) was examined. WI induced a prompt fall in MAP in all examined groups. This decrease of MAP was maximal after 1 h WI, showing a tendency to rise later on in pregnant women. Simultaneously a decrease of plasma renin activity (PRA), plasma aldosterone, AVP and an increase of ANF was noted. The WI induced endocrine reaction pattern was qualitatively similar, but quantitatively different in the examined groups. In contrast to the response of non-pregnant women, healthy pregnant women and women with EPH gestosis showed a significantly smaller increase in ANF secretion induced by WI. No correlation was found between PRA, plasma AVP, aldosterone and ANF respectively. In addition changes in PRA, aldosterone, AVP and ANF did not correlate with WI-induced changes in MAP. From data obtained in this paper it seems, that WI-induced MAP changes are not related significantly to changes of the above mentioned hormonal factors.