DNA methyltransferase inhibitors influence on the DIRAS3 and STAT3 expression and in vitro migration of ovarian and breast cancer cells

Objectives: Downregulation of DIRAS3 (DIRAS family, GTP-binding Ras-like 3) is related to ovarian and breast cancer progression. A possible mechanism that silences this gene is the promoter region DNA methylation. The potential reversibility of this epigenetic mechanism makes it more attractive candidate for new mode of cancer treatment. DIRAS3 regulates cell cycle, tumor dormancy and inhibits cancer cell growth and motility, all of which may indirectly depend on interaction with STAT3 (Signal Transducer and Activator of Transcription 3) classified as a potential oncogene. The restoration of DIRAS3 expression could inhibit cell proliferation and invasiveness. Material and methods: Human ovarian carcinoma cell line (A2780) and human breast cancer cell line (MCF7) were exposed to two DNA methyltransferase inhibitors (DNMTi): decitabine (5-aza-2’-deoxycytidine) [25 μM and 12.5 μM] and RG108 [150 μM and 100 μM]. In vitro migration changes of cancer cells were examined with wound healing assay. After 7 days of DNMTi treatment cells were harvested and DNA and RNA was isolated. The methylation status of the promoter sequences of DIRAS3 and STAT3 genes was determined using methylation specific PCR (MS-PCR). Level of target genes’ expression was quantified using quantitative reverse transcription PCR (QRT-PCR). Results and conclusions: The in vitro wound healing assay showed changes in the migration rate of both adherent cell lines after DNMTi treatment compared to the untreated cells. Relative balance between methylated and unmethylated variants of DIRAS3 after MS-PCR was shifted towards unmethylated version after DNMTi treatment in A2780 cells. Statistically significant dose dependent effect of decitabine and RG108 on DIRAS3 expression in A2780 cells was observed

Vitellogenins in the spider Parasteatoda tepidariorum – expression profile and putative hormonal regulation of vitellogenesis

Background: Knowledge about vitellogenesis in spiders is rudimentary. Therefore, the aim of study was to check the vitellogenin (Vg) presence in various tissues of the female spider Parasteatoda tepidariorum, determine when and where vitellogenesis starts and takes place, and the putative role of selected hormones in the vitellogenesis. Results: Here we show two genes encoding Vg (PtVg4 and PtVg6) in the genome of the spider P. tepidariorum. One gene PtVg4 and three subunits of Vg (250 kDa, 47 kDa and 30 kDa) are expressed in the midgut glands, ovaries and hemolymph. Heterosynthesis of the Vg in the midgut glands and autosynthesis in the ovaries were observed. Vitellogenesis begins in the last nymphal stage in the midgut glands (heterosynthesis). However, after sexual maturity is reached, Vg is also synthesized in the ovaries (autosynthesis). Changes in the PtVg4 expression level and in the Vg concentration after treatment with 20-hydroxyecdysone, a juvenile hormone analog (fenoxycarb) and an antijuvenoid compound (precocene I) were observed. Therefore, we propose a hypothetical model for the hormonal regulation of vitellogenesis in P. tepidariorum. Conclusions: Our results are the first comprehensive study on spider vitellogenesis. In our opinion, this work will open discussion on the evolutionary context of possible similarities in the hormonal control of vitellogenesis between P. tepidariorum and other arthropods as well as their consequences

Expression profile of genes encoding allatoregulatory neuropeptides in females of the spider Parasteatoda tepidariorum (Araneae, Theridiidae)

Allatoregulatory neuropeptides are multifunctional proteins that take part in the synthesis and secretion of juvenile hormones. In insects, allatostatins are inhibitors of juvenile hormone biosynthesis in the corpora allata while allatotropins, act as stimulators. By quantitative real-time PCR, we analyzed the gene expression of allatostatin A (PtASTA), allatostatin B (PtASTB), allatostatin C (PtASTC), allatotropin (PtAT) and their receptors (PtASTA-R, PtASTB-R, PtASTC-R, PtAT-R) in various tissues in different age groups of female spiders. In the presented manuscript, the presence of allatostatin A, allatostatin C, and allatotropin are reported in females of the spider P. tepidariorum. The obtained results indicated substantial differences in gene expression levels for allatoregulatory neuropeptides and their receptors in the different tissues. Additionally, the gene expression levels also varied depending on the female age. Strong expression was observed coinciding with sexual maturation in the neuroendocrine and nervous system, and to a lower extent in the digestive tissues and ovaries. Reverse trends were observed for the expression of genes encoding the receptors of these neuropeptides. In conclusion, our study is the first hint that the site of synthesis and secretion is fulfilled by similar structures as observed in other arthropods. In addition, the results of the analysis of spider physiology give evidence that the general functions like regulation of the juvenile hormone synthesis, regulation of the digestive tract and ovaries action, control of vitellogenesis process by the neuropeptides seem to be conserved among arthropods and are the milestone to future functional studies

Differential Quantitative Proteomics of Human Microvascular Endothelial Cells 1 by iTRAQ Reveals Palladin to be a New Biomarker During TGF-β1 Induced Endothelial Mesenchymal Transition

The study uses global quantitative proteomics to investigate the molecular mechanisms behind the induction of endothelial-mesenchymal transition (EndMT) by transforming growth factor–β (TGF-β). Orbitrap Velos mass spectrometers and iTRAQ – a labeling-based analysis were used to perform a global and quantitative comparison of two proteomes of Human Microvascular Endothelial Cells-1 (HMEC-1) treated or not treated by TGF-β1. iTRAQ analysis identified 43 differentially-expressed proteins in the early stages of EndMT induced by TGF-β1. From 5522 identified proteins, 26 were downregulated and 17 were upregulated, including proteins such as palladin, POTE I, torsin A and nucleoporin (NDC1). Further analysis of palladin revealed its increased mRNA and protein expression in response to TGF-β and Snail transcription factor. Our findings demonstrate that the newly- identified proteins may be involved in early stages of biological processes leading to EndM

Therapeutic treatment in the elderly with heart failure

Heart failure (HF) is a clinical syndrome that is the last common stage in many cardiovascular diseases. The main causes leading to the development of HF are, among others, primary impairment of myocardial contractility, volume and / or ventricular congestion, and obstruction of the diastolic duct due to pericardial diseases. The methods of detecting heart failure include: an accurate interview with the patient, laboratory tests and echocardiography. Treatment options include non-pharmacological, pharmacological and surgical treatment. A very important aspect of cure is also physical activity and appropriate nutritional treatment

Zwiększone stężenie białek ostrej fazy w płynie otrzewnowym kobiet z zaawansowaną endometriozą

Objectives: Most investigators agree that endometriosis is associated with a state of subclinical, non-infectious peritoneal inflammation. The objective of the study was to assess concentrations of two markers of the acute inflammatory phase proteins, haptoglobin and ceruloplasmin, in peritoneal fluid of endometriotic women. Material and methods: 229 women who underwent diagnostic or therapeutic laparoscopy were included in the study. Minimal, mild, moderate and severe endometriosis according to ASRM was confirmed in 119 women (study groups), whereas 110 patients suffered from simple serous or dermoid ovarian cysts (reference groups). Haptoglobin and ceruloplasmin concentrations in the peritoneal fluid samples aspirated during laparoscopy were measured using commercially available radial immunodiffusion kits. Results: The concentration of haptoglobin in the peritoneal fluid of women with endometriosis was significantly higher as compared to patients with serous and dermoid ovarian cysts. Significantly higher haptoglobin level was observed in patients with severe and moderate endometriosis as compared to women from both reference groups. No significant difference in the peritoneal fluid ceruloplasmin levels was found between patients with endometriosis and women from reference groups. However, it was noted that ceruloplasmin levels are higher in the subgroup of patients with severe endometriosis as compared to both reference groups and women with mild disease. Conclusions: Our results support the hypothesis that endometriosis is associated with subclinical inflammation within the peritoneal cavity. It may be speculated that pro-inflammatory stimuli strong enough to cause an increase in acute inflammatory phase proteins peritoneal fluid concentrations are observed only in the advanced stages of the disease.Cel pracy: Etiopatogeneza endometriozy nie jest do końca wyjaśniona, a większość badaczy zgadza się, że w płynie otrzewnowym kobiet chorych na tę chorobę stwierdza się subkliniczny proces zapalny pochodzenia nie-infekcyjnego. Celem pracy była ocena stężenia dwóch markerów, białek ostrej fazy zapalnej: haptoglobiny i ceruloplazminy w płynie otrzewnowym kobiet z endometriozą. Materiał i metody: Do badań włączono 229 kobiet, które przebyły diagnostyczną lub terapeutyczną laparoskopię. Endometriozę w stopniu minimalnym, łagodnym, umiarkowanym wg ASRM potwierdzono histologicznie u 119 kobiet (grupa badana). Pozostałe 110 pacjentek leczono z powodu prostych lub dermoidalnych torbieli (grupa referencyjna). Stężenia haptoglobiny i ceruloplazminy w próbkach płynu otrzewnowego oceniano przy użyciu komercyjnych zestawów do immunodyfuzji radialnej. Wyniki: Stężenie haptoglobiny w płynie otrzewnowym pacjentek z endometriozą było znacznie wyższe w porównaniu do pacjentek z grupy referencyjnej. Większe stężenie haptoglobiny dotyczyło podgrupy pacjentek z umiarkowaną i ciężką postacią choroby. Nie stwierdzono różnic w stężeniu ceruloplazminy w płynie otrzewnowym pacjentek z endometriozą w porównaniu do pacjentek z grupy referencyjnej. Jednakże wykazano, że stężenie ceruloplazminy jest większe w podgrupie pacjentek z ciężką postacią choroby w porównaniu do pacjentek z grupy referencyjnej i podgrupy pacjentek z łagodną postacią choroby. Wnioski: Wyniki naszych badań potwierdzają hipotezę o istnieniu subklinicznego stanu zapalnego w jamie otrzewnowej pacjentek z endometriozą. Wydaje się, że działanie silnych bodźców prozapalnych powodujących zwiększenie stężenia białek ostrej fazy w płynie otrzewnowym dotyczy jedynie pacjentek z zaawansowanymi postaciami choroby

Durvalumab Plus Carboplatin/Paclitaxel Followed by Maintenance Durvalumab With or Without Olaparib as First-Line Treatment for Advanced Endometrial Cancer: The Phase III DUO-E Trial

PURPOSE Immunotherapy and chemotherapy combinations have shown activity in endometrial cancer, with greater benefit in mismatch repair (MMR)-deficient (dMMR) than MMR-proficient (pMMR) disease. Adding a poly(ADP-ribose) polymerase inhibitor may improve outcomes, especially in pMMR disease. METHODS This phase III, global, double-blind, placebo-controlled trial randomly assigned eligible patients with newly diagnosed advanced or recurrent endometrial cancer 1:1:1 to: carboplatin/paclitaxel plus durvalumab placebo followed by placebo maintenance (control arm); carboplatin/paclitaxel plus durvalumab followed by maintenance durvalumab plus olaparib placebo (durvalumab arm); or carboplatin/paclitaxel plus durvalumab followed by maintenance durvalumab plus olaparib (durvalumab + olaparib arm). The primary end points were progression-free survival (PFS) in the durvalumab arm versus control and the durvalumab + olaparib arm versus control. RESULTS Seven hundred eighteen patients were randomly assigned. In the intention-to-treat population, statistically significant PFS benefit was observed in the durvalumab (hazard ratio [HR], 0.71 [95% CI, 0.57 to 0.89]; P = .003) and durvalumab + olaparib arms (HR, 0.55 [95% CI, 0.43 to 0.69]; P < .0001) versus control. Prespecified, exploratory subgroup analyses showed PFS benefit in dMMR (HR [durvalumab v control], 0.42 [95% CI, 0.22 to 0.80]; HR [durvalumab + olaparib v control], 0.41 [95% CI, 0.21 to 0.75]) and pMMR subgroups (HR [durvalumab v control], 0.77 [95% CI, 0.60 to 0.97]; HR [durvalumab + olaparib v control] 0.57; [95% CI, 0.44 to 0.73]); and in PD-L1-positive subgroups (HR [durvalumab v control], 0.63 [95% CI, 0.48 to 0.83]; HR [durvalumab + olaparib v control], 0.42 [95% CI, 0.31 to 0.57]). Interim overall survival results (maturity approximately 28%) were supportive of the primary outcomes (durvalumab v control: HR, 0.77 [95% CI, 0.56 to 1.07]; P = .120; durvalumab + olaparib v control: HR, 0.59 [95% CI, 0.42 to 0.83]; P = .003). The safety profiles of the experimental arms were generally consistent with individual agents. CONCLUSION Carboplatin/paclitaxel plus durvalumab followed by maintenance durvalumab with or without olaparib demonstrated a statistically significant and clinically meaningful PFS benefit in patients with advanced or recurrent endometrial cancer