Molecular Mechanisms of Lipopolysaccharide (LPS) Induced Inflammation in an Immortalized Ovine Luteal Endothelial Cell Line (OLENDO)

Escherichia coli (E. coli) is the most common Gram-negative bacterium causing infection of the uterus or mammary gland and is one of the major causes of infertility in livestock. In those animals affected by E. coli driven LPS-mediated infections, fertility problems occur in part due to disrupted follicular and luteal functionality. However, the molecular mechanisms by which LPS induces inflammation, and specifically, the role of LPS in the disruption of capillary morphogenesis and endothelial barrier function remain unclear. Here, we hypothesized that LPS may lead to alterations in luteal angiogenesis and vascular function by inducing inflammatory reactions in endothelial cells. Accordingly, OLENDO cells were treated with LPS followed by evaluation of the expression of selected representative proinflammatory cytokines: NF-kB, IL6, IL8, TNFα, and ICAM 1. While TNFα was not affected by treatment with LPS, transcripts of NF-kB, IL6, and IL8 were affected in a dosage-dependent manner. Additionally, the activity of TLR2 and TLR4 was blocked, resulting in suppression of the LPS-induced expression of ICAM 1, NF-kB, IL6, and IL8. Inhibition of the PKA or MAPK/ERK pathways suppressed the LPS-stimulated expression of NF-kB, IL6, and IL8, whereas blocking the PKC pathway had the opposite effect. Furthermore, LPS-induced phosphorylation of Erk1 and Erk2 was inhibited when the TLR4 or MAPK/ERK pathways were blocked. Finally, LPS seems to induce inflammatory processes in OLENDO cells via TLR2 and TLR4, utilizing different signaling pathways

Determination of novel reference genes for improving gene expression data normalization in selected canine reproductive tissues – a multistudy analysis

Background Real time RT-PCR (qPCR) is a useful and powerful tool for quantitative measurement of gene expression. The proper choice of internal standards such as reference genes is crucial for correct data evaluation. In female dogs, as in other species, the reproductive tract is continuously undergoing hormonal and cycle stage-dependent morphological changes, which are associated with altered gene expression. However, there have been few attempts published so far targeted to the dog aimed at determining optimal reference genes for the reproductive organs. Most of these approaches relied on genes previously described in other species. Large-scale transcriptome-based experiments are promising tools for defining potential candidate reference genes, but were never considered in this context in canine research. Results Here, using available microarray and RNA-seq datasets derived from reproductive organs (corpus luteum, placenta, healthy and diseased uteri) of dogs, we have performed multistudy analysis to identify the most stably expressed genes for expression studies, in each tissue separately and collectively for different tissues. The stability of newly identified reference genes (EIF4H, KDELR2, KDM4A and PTK2) has been determined and ranked relative to previously used reference genes, i.e., GAPDH, β-actin and cyclophillin A/PPIA, using RefFinder and NormFinder algorithms. Finally, expression of selected target genes (luteal IL-1b and MHCII, placental COX2 and VEGFA, and uterine IGF2 and LHR) was re-evaluated and normalized. All proposed candidate reference genes were more stable, ranked higher and introduced less variation than previously used genes. Conclusions Based on our analyses, we recommend applying KDM4A and PTK2 for normalization of gene expression in the canine CL and placenta. The inclusion of a third reference gene, EIF4H, is suggested for healthy uteri. With this, the interpretation of qPCR data will be more reliable, allowing better understanding of canine reproductive physiology

Antigestagens Mediate the Expression of Decidualization Markers, Extracellular Matrix Factors and Connexin 43 in Decidualized Dog Uterine Stromal (DUS) Cells

Feto-maternal communication in the dog involves the differentiation of stromal cells into decidual cells. As the only placental cells expressing the nuclear progesterone (P4) receptor (PGR), decidual cells play crucial roles in the maintenance and termination of pregnancy. Accordingly, to investigate possible PGR-mediated mechanisms in canine decidual cells, in vitro decidualized dog uterine stromal (DUS) cells were treated with functional PGR-blockers, mifepristone and aglepristone. Effects on decidualization markers, epithelial and mesenchymal factors, and markers of cellular viability were assessed. Decidualization increased the expression of PTGES, PGR, IGF1, and PRLR, along with ECM1, COL4 and CX43, but downregulated IGF2. DUS cells retained their mesenchymal character, and the expression of COL4 indicated the mesenchymal-epithelial transformation. Antigestagen treatment decreased the availability of PTGES, PRLR, IGF1 and PGR. Furthermore, antigestagens decreased the mRNA and protein expression of CX43, and transcriptional levels of ECM1 and COL4. Additionally, antigestagens increased levels of activated-CASP3 (a proapoptotic factor), associated with lowered levels of PCNA (a proliferation marker). These data reveal important aspects of the functional involvement of PGR in canine decidual cells, regarding the expression of decidualization markers and acquisition of epithelial-like characteristics. Some of these mechanisms may be crucial for the maintenance and/or termination of canine pregnancy

Motywy inwestycji zagranicznych polskich przedsiębiorstw

The aim of this study was to examine the motives of foreign direct investment (FDI) of Polish enterprises. Our studies have confirmed that the market seeking and – to a lesser extent – resource seeking motives are most important for Polish companies investing abroad. The search for efficiency is still not as significant as above factors. The study also shown the importance of geographical distance – this is reflected by concentration of Polish FDI in neighboring countries. We also identified the differences in motives to invest across various sectors and equity based entry modes.Celem tej pracy było zbadanie motywów bezpośrednich inwestycji zagranicznych (BIZ) polskich przedsiębiorstw. Badania pozwoliły na stwierdzenie, iż głównymi motywami BIZ polskich firm było poszukiwanie rynków zbytu oraz – w mniejszym stopniu – zasobów. Czynniki związane z poszukiwaniem efektywności nie miały istotnego znaczenia. Wykazano też znaczenia dystansu geograficznego i koncentrację polskich BIZ w krajach sąsiedzkich. Wskazano również na różnice w motywach inwestowania pomiędzy sektorami usługowymi i produkcyjnymi oraz transgranicznymi przejęciami i inwestycjami od podstaw

Transcriptomic profiling of canine decidualization and effects of antigestagens on decidualized dog uterine stromal cells

Maternal-stroma derived decidual cells, the only cell population in the canine placenta expressing the nuclear progesterone (P4) receptor (PGR), are crucial for the maintenance of canine pregnancy. Decreased circulating progesterone (P4) levels, or blockage of PGR function with antigestagens, terminate canine pregnancy. As an in vitro model for canine decidualization, dog uterine stromal (DUS) cells can be decidualized in vitro with cAMP. The antigestagens aglepristone and mifepristone ablate the expression of decidualization markers in DUS cells (e.g., PGR, PRLR, IGF1 or PTGES). Here, the transcriptome profile of DUS cells was investigated to acquire deeper insights into decidualization-associated changes. Additionally, effects mediated by antigestagens (competitive PGR blockers) in decidualized cells were assessed. Decidualization led to the upregulation of 1841 differentially expressed genes (DEGs, P and FDR < 0.01) involved in cellular proliferation and adhesion, mesenchymal-epithelial transition, extracellular matrix organization, and vaso- and immunomodulation. The 1475 DEGs downregulated after decidualization were mostly associated with apoptosis and cell migration. In decidualized DUS cells, aglepristone modulated 1400 DEGs and mifepristone 1558 DEGs. Interestingly, around half of the identified DEGs were modulated by only one of the antigestagens. In all cases, however, PGR-blockage was mainly associated with an inversion of several decidualization-induced effects. Comparison between antigestagen-mediated effects and transcriptional changes in the canine placenta at term allowed the identification of 191 DEGs associated with diminished cell proliferation and adhesion, and vascular and immune modulation. This study emphasizes the importance of P4/PGR signaling for decidual cell function, providing new insights into the maintenance of canine pregnancy

Surgical Repair of Postinfarction Ventricular Septal Rupture: Systematic Review and Meta-Analysis.

Background. Ventricular septal rupture (VSR) is a rare but life-threatening complication after acute myocardial infarction. Although surgical correction is challenging and associated with high mortality, it remains the treatment of choice. This systematic review and meta-analysis aimed to evaluate the early outcome of surgical VSR repair.Methods. We searched electronic databases from January 1998 to February 2020. Studies reporting patients undergoing surgical treatment for VSR were analyzed. The primary outcome assessed was operative mortality. Differences were expressed as odds ratios (ORs) with 95% confidence intervals (CIs) to assess the relationships of predefined surgical variables and clinical prognosis.Results. A total of 6361 adult patients from 41 studies were identified. Operative mortality was 38.2%. Pooled ORs showed increased odds of operative mortality in patients with preoperative or perioperative intraaortic balloon pump insertion (OR = 3.48; 95% CI, 3.01-4.02; P >= .001), right ventricular dysfunction (OR = 2.85; 95% CI, 1.47-5.52; P = .002), posterior VSR (OR = 1.73; 95% CI, 1.30-2.31; P >= .001), and emergency surgery (OR = 3.79; 95% CI, 2.52-5.72; P >= .001). Temporal trend evaluation revealed no difference over time in the operative mortality rate; it was 34% in both time-related groups (1971-2000 versus 2001-2018).Conclusions. Ventricular septal rupture repair has a high operative mortality. Patients with preoperative or perioperative intraaortic balloon pump support, right ventricular dysfunction at presentation, or posterior defects, and those undergoing emergent VSR correction have increased odds of operative mortality. (C) 2021 by The Society of Thoracic Surgeons. Published by Elsevier Inc

Expression patterns of intestinal calcium transport factors and ex-vivo absorption of calcium in horses

BACKGROUND: In many species, the small intestine is the major site of calcium (Ca(2+)) absorption. The horse differs considerably from most other species with regard to the physiology of its Ca(2+) metabolism and digestion. Thus, this study was performed to get more information about the transcellular Ca(2+) absorption in the horse.Two mechanisms of intestinal Ca(2+) absorption are described: the passive paracellular pathway and the active, vitamin D-dependent transcellular pathway. The latter involves the following elements: vitamin D receptors (VDR), transient receptor potential vanilloid channel members 5 and 6 (TRPV5/6), calbindin-D9k (CB), the Na/Ca exchanger (NCX1) and the plasma membrane Ca-ATPase (PMCA). The aim of the present study was to investigate the protein and mRNA expression patterns of VDR, CB and TRPV6 and the ex-vivo Ca(2+) absorption in horses, assessed by qualitative and quantitative RT-PCR, western blot, immunohistochemistry and the Ussing chamber technique. RESULTS: Highest CB and TRPV6 mRNA levels were detected in the duodenum as compared to the middle parts of the jejunum and ileum and several sites of the large intestine. VDR mRNA levels did not change significantly throughout the intestine. TRPV5 mRNA was not detectable in the horse intestine. The highest VDR and CB protein levels were measured in the duodenum. Ussing chamber studies revealed ex-vivo Ca(2+) absorption only in the duodenum, but not in cecum and specific sites of the colon. CONCLUSION: The present findings suggest that TRPV6, CB and VDR may be involved in active intestinal Ca(2+) absorption in horses, as described for other mammals. TRPV5 may not play a major role in this process. Furthermore, the expression patterns of these Ca(2+) transport elements and the results of the Ussing chamber procedure indicate that a significant part of active intestinal Ca(2+) absorption occurs in the duodenum in this species

Kaniulacja żyły i tętnicy szyjnej do krążenia pozaustrojowego podczas operacji z minitorakotomii u pacjenta obciążonego bez możliwości klasycznego dostępu

Minimally-invasive surgery is more and more often considered to be an advisable treatment in case of patients with mitral or triscupid valve dysfunctions, as well as those with heart tumor. The incision is reduced to minimum. As a result, the blood loss, postoperative pain and cardiorespiratory complications are also reduced. Minimally-invasive techniques are being developed dynamically all over the world, although they are not available yet in many medical centres. In this article, a case of a female patient in extremely severe condition is described. The women had been diagnosed with left atrial myxoma and underwent a minimally-invasive surgery with alternative access used for the purpose of extracorporeal circulation.Minimally-invasive surgery is more and more often considered to be an advisable treatment in case of patients with mitral or triscupid valve dysfunctions, as well as those with heart tumor. The incision is reduced to minimum. As a result, the blood loss, postoperative pain and cardiorespiratory complications are also reduced. Minimally-invasive techniques are being developed dynamically all over the world, although they are not available yet in many medical centres. In this article, a case of a female patient in extremely severe condition is described. The women had been diagnosed with left atrial myxoma and underwent a minimally-invasive surgery with alternative access used for the purpose of extracorporeal circulation

safety and efficacy of miniaturized extracorporeal circulation when compared with off pump and conventional coronary artery bypass grafting evidence synthesis from a comprehensive bayesian framework network meta analysis of 134 randomized controlled trials involving 22 778 patients

Coronary artery bypass grafting (CABG) remains the standard of care in patients with extensive coronary artery disease. Yet the use of cardiopulmonary bypass (CPB) is believed to be a major determinant of perioperative morbidity. Novel techniques are sought to tackle the shortcomings of CPB, among them off-pump coronary artery bypass (OPCAB) and miniaturized extracorporeal circulation (MECC) systems have been extensively tested in randomized controlled trials (RCTs). To assess perioperative safety and efficacy of MECC and OPCAB when compared with conventional extracorporeal circulation (CECC).Published literature and major congress proceedings were screened for RCTs evaluating the safety and efficacy of MECC, OPCAB and CECC. Selected end-points such as 30-day all-cause mortality, myocardial infarction (MI), cerebral stroke, postoperative atrial fibrillation (POAF) and renal dysfunction were assessed in a Bayesian-framework network meta-analysis.A total of 134 studies with 22 778 patients were included. When compared with CECC, both OPCAB and MECC significantly reduced 30-day all-cause mortality [odds ratios (95% credible intervals): 0.75 (0.51-0.99) and 0.46 (0.22-0.91)], respectively. No differences in respect to MI were demonstrated with either strategy. OPCAB, when compared with CECC, reduced the odds of cerebral stroke [0.57 (0.34-0.80)]; 60% reduction was observed with MECC when compared with CECC [0.40 (0.19-0.78)]. Both OPCAB and MECC reduced the odds of POAF [0.66 (0.48-0.90) and 0.62 (0.35-0.98), respectively] when compared with CECC. OPCAB conferred over 30% reduction of renal dysfunction when compared with CECC [0.69 (0.46-0.92)]. MECC reduced these odds by more than 50% [0.47 (0.24-0.89)]. Ranking of treatments emerging from the probability analysis (highest to lowest SUCRA values) was MECC followed by OPCAB and CECC.MECC and OPCAB both improve perioperative outcomes following coronary bypass surgery when compared with conventional CABG performed with extracorporeal circulation. MECC may represent an attractive compromise between OPCAB and CECC