Ninety-six–hour starved peripheral blood mononuclear cell supernatant inhibited LA7 breast cancer stem cells induced tumor via reduction in angiogenesis and alternations in Gch1 and Spr expressions

Introduction The microenvironment of solid tumors such as breast cancer is heterogeneous and complex, containing different types of cell, namely, cancer stem cells and immune cells. We previously reported the immunoregulatory behavior of the human immune cell in a solid tumor microenvironment-like culture under serum starvation stress for 96 h. Here, we examined the effect of this culture-derived solution on breast cancer development in rats. Method Ninety-six–hour starved PBMCs supernatant (96 h-SPS) was collected after culturing human PBMCs for 96 h under serum starvation condition. Breast cancer stem cells, LA7 cell line, was used for in vitro study by analyzing gene expression status and performing cytotoxicity, proliferation, scratch wound healing assays, followed by in vivo tumor induction in three groups of mature female Sprague Dawley rats. Animals were treated with 96 h-SPS or RPMI and normal saline as control, n = 6 for each group. After biochemical analysis of iron, lactate, and pH levels in the dissected tumors, Ki67 antigen expression, angiogenesis, and necrosis evaluation were carried out. Metabolic-related gene expression was assessed using RT-qPCR. Moreover, 96 h-SPS composition was discovered by Nano-LC-ESI-MS/MS. Results 96 h-SPS solution reduced the LA7 cell viability, proliferation, and migration and Gch1 and Spr genes expression in vitro ( p < 0.05), whereas stemness gene Oct4 was upregulated ( p < 0.01). The intracellular lactate was significantly decreased in the 96 h-SPS treated group ( p = 0.007). In this group, Gch1 and Spr were significantly downregulated ( p < 0.05), whereas the Sox2 and Oct4 expression was not changed significantly. The number of vessels and mitosis (Ki67 + cells) in the 96 h-SPS–treated group was significantly reduced ( p = 0.024). The increased rate of necrosis in this group was statistically significant ( p = 0.04). Last, proteomics analysis revealed candidate effectors’ components of 96 h-SPS solution. Conclusion 96 h-SPS solution may help to prevent cancer stem cell mediated tumor development. This phenomenon could be mediated through direct cytotoxic effects, inhibition of cell proliferation and migration in association with reduction in Gch1 and Spr genes expression, angiogenesis and mitosis rate, and necrosis augmentation. The preliminary data obtained from the present study need to be investigated on a larger scale and can be used as a pilot for further studies on the biology of cancer development

Голодомор 1932 –– 1933 рр. в Україні як геноцид

Given its fundamental role in development and cancer, the Wnt-beta-catenin signaling pathway is tightly controlled at multiple levels. RING finger protein 43 (RNF43) is an E3 ubiquitin ligase originally found in stem cells and proposed to inhibit Wnt signaling by interacting with the Wnt receptors of the Frizzled family. We detected endogenous RNF43 in the nucleus of human intestinal crypt and colon cancer cells. We found that RNF43 physically interacted with T cell factor 4 (TCF4) in cells and tethered TCF4 to the nuclear membrane, thus silencing TCF4 transcriptional activity even in the presence of constitutively active mutants of beta-catenin. This inhibitory mechanism was disrupted by the expression of RNF43 bearing mutations found in human gastrointestinal tumors, and transactivation of the Wnt pathway was observed in various cells and in Xenopus embryos when the RING domain of RNF43 was mutated. Our findings indicate that RNF43 inhibits the Wnt pathway downstream of oncogenic mutations that activate the pathway. Mimicking or enhancing this inhibitory activity of RNF43 may be useful to treat cancers arising from aberrant activation of the Wnt pathwa

Diagnosis of Helicobacter pylori: Changes towards the Future

Since the first evidence demonstrating the dramatically high incidence of H. pylori infection and the subsequent medical challenges it incurs, health management of H. pylori infection has been a high priority for health authorities worldwide. Despite a decreasing rate of infection in western countries, prevalence of H. pylori infection in developing and in some industrial countries is still very high. Whereas treatment and vaccination against H. pylori is a contemporary issue in medical communities, selective treatment and prior high-throughput screening of the subject population is a major concern of health organizations. So far, diagnostic tests are either elaborative and require relatively advanced medical care infrastructure or they do not fulfill the criteria recommended by the Maastricht IV/Florence consensus report. In this review, in light of recent scientific studies, we highlight current and possible future approaches for the diagnosis of H. pylori. We point out that novel non-invasive tests may not only cover the requirements of gold standard methods in H. pylori detection but also offer the potential for risk stratification of infection in a high throughput manner

Evaluation of weed control methods on sugar beet (Beta vulgaris L.) yield at different levels of nitrogen

Introduction: Weed competition is one of the major factors which limit sugar beet production in the world. Weed – crop interactions are based on competition for water, nutrients and light and allelopathic effects may also play a small role. In sugar beet weed interference, all these factors are important too, but the light is of prime importance. Due to the fact that a lot of weeds can grow above the sugar beet canopy and reduce the amount of photosynthetic radiation reaching the crop, these weeds are stronger competitors compared to smaller weeds. In much sugar beet growing areas dicot weeds of the families Chenopodiaceae, Asteraceae, Brassicaceae and Polygonaceae are of major importance. The monocots are less important compared to dicot weeds. Competition from uncontrolled annual weeds that emerge within 8 weeks of sowing or within 4 weeks of the crop reaching the two-leaf stage can reduce root yields by 26–100% .Weeds that emerge 8 weeks after sowing, and particularly after the sugar beet plants have eight or more leaves, are less likely to affect yield. Although tractor hoeing and hand labour are still used in many production areas, herbicides have been the primary method of weed control in sugar beet. The effectiveness of pre-emergence residual herbicides decreases with reductions in rainfall or soil wet content. Therefore, less than 10 % of the total sugar beet crop is treated with pre-emergence herbicides. The remaining 90 % depends solely on a selection of post-emergence herbicides to maintain season-long weed control. The major herbicides are phenmedipham, chloridazon, metamitron. Mixtures of post-emergence, broad spectrum herbicides have to be applied to control the wide range of weed species in sugar beet crops. Materials and Methods: To study the effects of weeds control by hand weeding and herbicides combination with two selective herbicides at different levels of nitrogen application on sugar beet yield and quality characteristics, an experiment carried out in TorbateJam Township as statistical design with split plots in a randomized complete block design with three replications during 2014. Experiment treatments included, the main factor involving four levels of different Nitrogen application (0, 100. 150 and 200 Kg.ha-1), sub factor involving combination of chloridazon + phenmedipham and metamitron + phenmedipham at 5 Kg.ha-1 herbicides. Four weeks after treatments, sampling of weeds and sugar beet carried out in middle of the plots with 0.5 × 0.5 quadrate. Then, samples were dried at oven-dried at 75 °C for 48 h and weighed. At the final harvest, to determine the grade, amino nitrogen, sodium, potassium with Betalyzr at sugar sector of Agricultural Center laboratory, sampling was removed from the middle of each plot. Results Discussion: The results showed that application of nitrogen fertilizer and herbicide treatments were significantly different from each other at 1% and 5% levels, respectively. Based on experiment results, the highest root yield of sugar beet was obtained hand weeding with 200 kg N.ha-1 treatments. In between treatments of weed chemical control, metamitron + phenmedipham herbicides with 200 kg N.ha-1 was showed the highest root yield of sugar beet. High net sugar beet yield also was obtained at complete weed control with 200 kg N.ha-1, and metamitron + phenmedipham herbicides application with 200 kg N.ha-1 treatments. Also, the highest net and gross sugars were obtained at without weed control + 0 kg N.ha-1 treatments. Conclusion: In conclusion, According to results of this study root yield and net and gross sugar were increased by increasing 200 kg nitrogen per hectare. Also, the highest net and gross sugar yield related to the using of hand weeding and combination herbicides of metamitron + phenmedipham and chloridazon + phenmedipham and application of 200 kg.ha-1 nitrogen fertilizer with weed control as well. Between weed controls treatments, root yield were increased by hand weeding compared to herbicide application and between herbicide treatments at Nitrogen different levels, by using of metamitron + phenmedipham than chloridazon + phenmedipham. Between chemical treatments, net and gross sugar yield, shoot dry weight and dry matter yield of sugar beet were more in combination of metamitron + phenmedipham than chloridazon + phenmedipham. While, weeds density and biomass were lesser in herbicide combination of metamitron + phenmedipham compared to chloridazon + phenmedipham. On the other hand, among treatments interaction, the highest root yield and percent of sugar and pure sugar were obtained by 200 kg per hectare Nitrogen fertilizer accompanied weeds control and without weeds control with lack of Nitrogen application respectively

H. pylori Virulence Factors: Influence on Immune System and Pathology

Helicobacter pylori is the most widespread chronic bacterial agent in humans and is well recognized for its association with ulcer disease and gastric cancer, with both representing major global health and socioeconomic issues. Given the high level of adaptation and the coevolution of this bacterium with its human host, a thorough and multidirectional view of the specific microbiological characteristics of this infection as well as the host physiology is needed in order to develop novel means of prevention of therapy. This review aims to pinpoint some of these potentially important angles, which have to be considered mutually when studying H. pylori's pathogenicity. The host's biological changes due to the virulence factors are a valuable pillar of H. pylori research as are the mechanisms by which bacteria provoke these changes. In this context, necessary adhesion molecules and significant virulence factors of H. pylori are discussed. Moreover, metabolism of the bacteria, one of the most important aspects for a better understanding of bacterial physiology and consequently possible therapeutic and prophylactic strategies, is addressed. On the other hand, we discuss the recent experimental proofs of the “hygiene hypothesis” in correlation with Helicobacter's infection, which adds another aspect of complexity to this infection

Influence of HBgGT on the activation of the NF-κB, AP-1 and CREB pathways.

<p>NF-κB, AP-1 and CREB transcriptional activity in transiently transfected HCT116 (A), and DLD-1 and LS174T (B) cells. Recombinant HBgGT (5µg/ml) was added to <i>H. bilis</i> ΔgGT-infected cells and transcriptional activity determined after 24 hours of infection. Results are expressed as mean of relative luciferase activity to renilla of three independent experiments, normalized to the untreated control. *p<0.05, **p<0.005, ***p<0.0005. Asterisks on top of bars indicate significance relative to untreated control; asterisks on bars indicate significance level between indicated conditions.</p

Influence of glutamine supplementation on IL-8 secretion by <i>H. bilis</i> co-cultures and HBgGT-treated cells.

<div><p>A) IL-8 production by HCT116 and DLD-1 cells. <i>H. bilis</i> (MOI 50) infected cells were supplemented with 3mM L-glutamine (Gln). Supernatants of 24 hour treated cells were collected and IL-8 secretion determined by ELISA. Results are expressed as mean of three independent experiments. *p<0.05, **p<0.005, ***p<0.0005.</p> <p>B) IL-8 levels secreted by HCT116 and DLD cells after glutamine supplementation (Gln 2mM, where indicated) of HBgGT PIM (pre-incubated medium, 5µg HBgGT/ml of cell culture medium). Heat-inactivated HBgGT PIM (inactive HBgGT pre-incubated at 5µg/ml of culture medium) was used as an enzymatically inactive control. Results are expressed as mean of three independent experiments. *p<0.05, **p<0.005, ***p<0.0005.</p></div