Analysis of ANRIL gene polymorphism rs4977574 association with kidney cancer development in Ukrainian population.

ANRIL (Antisense Non-coding RNA in the INK4 Locus, also known as CDKN2B-AS1) – 3.8-kb long non-coding RNA transcribed from the antisense strand of INK4b-ARF-INK4a gene cluster. It is known that ANRIL overexpression is associated with development of oncological pathologies of different localization. In addition, there are a number of studies devoted to role of ANRIL genetic polymorphism in emergence and progression of tumors, including tumors of genitourinary system. The aim of the study was to check the possible association between ANRIL gene polymorphism rs4977574 and kidney cancer development in representatives of Ukrainian population. Whole venous blood of 101 patients with clear cell renal cell carcinoma (CCRCC) (42 women and 59 men) and 100 patients without oncology history (34 women and 66 men) was used in the study. DNA from blood white cells was extracted using GeneJET Whole Blood Genomic DNA Purification Mini Kit (Thermo Fisher Scientific, USA). Genotyping of rs4977574 ANRIL gene polymorphic locus was performed using real-time polymerase chain reaction (real-time PCR) method in the presence of TaqMan assay C_31720978_30. The mathematical data were processed using the SPSS software package (version 17.0). P values <0.05 were considered as statistically significant. It was found that difference in rs4977574-genotype distribution between patients with CCRCC and control persons was absent in general group (P=0.216). At the same time, the statistical analysis stratified by gender showed that both in female and male subjects rs4977574-genotypes frequency also did not differ significantly between comparison groups (P=0.526 and P=0.160, respectively). However, after adjusting for age, body mass index, and smoking habits statistically significant association between rs4977574 ANRIL gene polymorphism and risk of kidney cancer development was detected in male subjects under superdominant inheritance model (P=0.049). It was revealed that heterozygotes (AG-genotype) have 2.17-fold  higher risk of CCRCC development (95% CI=1.005-4.695) compared to patients with AA- and GG-genotypes. In summary, this is the first report about ANRIL gene polymorphisms association with kidney cancer. Obtained results revealed that rs4977574 is related to kidney cancer risk only in Ukrainian men. Male individuals with AG-genotype have higher risk of CCRCC development compared to AA- and GG-genotypes carriers

Synergism between the N-acetyltransferase 2 gene and oxidant exposure increases the risk of idiopathic male infertility

N-acetyltransferase (NAT2) is a phase-II xenobiotic-metabolizing enzyme participating in the detoxification of toxic arylamines, aromatic amines and hydrazines. The present study was designed to investigate whether two common single-nucleotide polymorphisms (SNP) of the NAT2 gene (481C>T, rs1799929; 590G>A, rs1799930) are associated with susceptibility to idiopathic male infertility and to assess if the risk is modified by oxidant and antioxidant exposures. A total 430 DNA samples (203 infertile patients and 227 fertile men) were genotyped for the polymorphisms by PCR and restriction fragment length polymorphism. No association was found between the NAT2 polymorphisms and idiopathic male infertilit

Analysis of association between long non-coding RNA HOTAIR gene rs1899663 polymorphism and disease-free survival in kidney cancer patients

The aim of the current study was to investigate the possible association of HOTAIR gene rs1899663 polymorphism with kidney cancer patients survival and clinicopathological characteristics of kidney cancer. Methods. The whole venous blood of 101 patients with clear cell renal cell carcinoma (CCRCC)  (42 women and 59 men) was used in the study. Genotyping of rs1899663 HOTAIR gene polymorphic locus was performed by polymerase chain reaction followed by restriction fragment length polymorphism analysis (PCR-RFLP). Statistical analysis was performed using SPSS (version 17.0). The Kaplan-Meier test and the Cox regression were used to test the possible association between  rs1899663 polymorphism of long non-coding RNA HOTAIR gene and the age of CCRCC onset. P values ​​< 0.05 were considered as statistically significant. Results. The results of HOTAIR gene rs1899663 polymorphism genotyping showed that the ratio of GG-homozygotes, GT-heterozygotes and TT-homozygotes in CCRCC patients was 39.6%, 52.5%, and 7.9%, respectively. This distribution did not deviate from the expected by Hardy-Weinberg law (P = 0.143). Results of one-way ANOVA showed that rs1899663-locus of HOTAIR gene was not associated with tumor size of CCRCC patients (P > 0.05), was not related to body mass index, erythrocyte sedimentation rate, fasting glucose, hemoglobin, creatinine and leukocyte amount in the blood of CCRCC patients (P > 0.05). In addition, the results of the Kaplan-Meyer test showed that life expectancy until the CCRCC occurrence does not depend on rs1899663 SNP (log-rank P = 0.739). At the same time, the results of Cox regression analysis both before and after adjusting for covariates (sex, body mass index, metastases, smoking habits and alcohol abuse) showed that CCRCC risk development with age does not depend on HOTAIR gene rs1899663 polymorphism (P > 0.05). Conclusions. This is the first report about the possible link between HOTAIR gene polymorphism and survival of kidney cancer patients both in Ukraine and worldwide. The rs1899663 polymorphic site of HOTAIR gene is not associated with the age of CCRCC onset in the Ukrainian population. In addition, the rs1899663 SNP is also not related to tumor size and clinical фтв laboratory data in patients with kidney cancer

Thr83ala Gene Polymorphism Association with Arterial Calcification, Acute Coronary Syndrome and Ischemic Strokes in Older Adults

BACKGROUND: Calcification of the arteries of the lower extremities is a very common pathological process, which has an independent significance in the development of cardiovascular diseases. There is evidence that development of calcification of the arteries correlates with a mutation of the MGP protein gene representing by the Thr83Ala polymorphism.AIM: The purpose of the study was to analyse the connection of Thr83Ala polymorphism of the MGP gene with the development of calcification of the arteries of the lower extremities.METHODS: The study involved 80 patients. Half of them had signs of calcification of the arteries of the lower extremities. The allelic Thr83Ala polymorphism of the MGP protein gene was determined by polymerase chain reaction, establishing the presence of calcification of the arteries by radiological and dopplerographic methods.RESULTS: The study aimed to analyse the association of the Thr83Ala polymorphism of the matrix Gla protein gene with the development of calcification of the arteries of the lower extremities. The data obtained suggest that the replacement of threonine by alanine in the 83rd position of the MGP molecule can affect the functional properties of the protein and in particular its anticarcinogenic properties. Although there was no difference in the distribution of different variants of the genotype by Thr83Ala to the MGP gene polymorphism in patients with CA and healthy patients, but in the distribution of genotypes in the comparison groups separated by sex, it was found that in women, carriage of the Ala allele in a homozygous state is a factor, which protects the development of arterial calcination in the elderly and senile.CONCLUSION: Differences in the distribution of different variants of the genotype according to Thr83Ala to the polymorphism of the MGP gene between patients with CAD and healthy patients do not exceed the limits of statistical significance. In the distribution of genotypes in the comparison groups divided by sex, it was found that in women the carrier of the Ala-allele in the homozygous state is a factor that prevents the development of Menkeberg arteriosclerosis in the elderly and old age

Selective Remodeling: Refining Neural Connectivity at the Neuromuscular Junction

A primer on new research by Fuentes-Medel and colleagues explains the important role of non-neural cells in clearing neural debris, which is continuously produced during the normal remodeling processes that establish and maintain neural connectivity

Complementarity, quantum erasure and delayed choice with modified Mach-Zehnder interferometers

Often cited dictums in Quantum Mechanics include "observation disturbance causes loss of interference" and "ignorance is interference". In this paper we propose and describe a series of experiments with modified Mach-Zehnder interferometers showing that one has to be careful when applying such dictums. We are able to show that without interacting in any way with the light quantum (or quanta) expected to behave "wave-like", interference fringes can be lost by simply gaining (or having the potential to gain) the which-path knowledge. Erasing this information may revive the interference fringes. Delayed choice can be added, arriving to an experiment in line with Wheeler's original proposal. We also show that ignorance is not always synonym with having the interference fringes. The often-invoked "collapse of the wavefunction" is found to be a non-necessary ingredient to describe our experiments.Comment: 8 pages, 3 figures; to appear in EPJ

Antigen-Displaying Lipid-Enveloped PLGA Nanoparticles as Delivery Agents for a Plasmodium vivax Malaria Vaccine

The parasite Plasmodium vivax is the most frequent cause of malaria outside of sub-Saharan Africa, but efforts to develop viable vaccines against P. vivax so far have been inadequate. We recently developed pathogen-mimicking polymeric vaccine nanoparticles composed of the FDA-approved biodegradable polymer poly(lactide-co-glycolide) acid (PLGA) “enveloped” by a lipid membrane. In this study, we sought to determine whether this vaccine delivery platform could be applied to enhance the immune response against P. vivax sporozoites. A candidate malaria antigen, VMP001, was conjugated to the lipid membrane of the particles, and an immunostimulatory molecule, monophosphoryl lipid A (MPLA), was incorporated into the lipid membranes, creating pathogen-mimicking nanoparticle vaccines (VMP001-NPs). Vaccination with VMP001-NPs promoted germinal center formation and elicited durable antigen-specific antibodies with significantly higher titers and more balanced Th1/Th2 responses in vivo, compared with vaccines composed of soluble protein mixed with MPLA. Antibodies raised by NP vaccinations also exhibited enhanced avidity and affinity toward the domains within the circumsporozoite protein implicated in protection and were able to agglutinate live P. vivax sporozoites. These results demonstrate that these VMP001-NPs are promising vaccines candidates that may elicit protective immunity against P. vivax sporozoites.United States. Dept. of Defense (contract W911NF-07-D-0004)Ragon Institute of MGH, MIT and Harvar

LEM-3 – A LEM Domain Containing Nuclease Involved in the DNA Damage Response in C. elegans

The small nematode Caenorhabditis elegans displays a spectrum of DNA damage responses similar to humans. In order to identify new DNA damage response genes, we isolated in a forward genetic screen 14 new mutations conferring hypersensitivity to ionizing radiation. We present here our characterization of lem-3, one of the genes identified in this screen. LEM-3 contains a LEM domain and a GIY nuclease domain. We confirm that LEM-3 has DNase activity in vitro. lem-3(lf) mutants are hypersensitive to various types of DNA damage, including ionizing radiation, UV-C light and crosslinking agents. Embryos from irradiated lem-3 hermaphrodites displayed severe defects during cell division, including chromosome mis-segregation and anaphase bridges. The mitotic defects observed in irradiated lem-3 mutant embryos are similar to those found in baf-1 (barrier-to-autointegration factor) mutants. The baf-1 gene codes for an essential and highly conserved protein known to interact with the other two C. elegans LEM domain proteins, LEM-2 and EMR-1. We show that baf-1, lem-2, and emr-1 mutants are also hypersensitive to DNA damage and that loss of lem-3 sensitizes baf-1 mutants even in the absence of DNA damage. Our data suggest that BAF-1, together with the LEM domain proteins, plays an important role following DNA damage – possibly by promoting the reorganization of damaged chromatin

Differential Modulation of TCF/LEF-1 Activity by the Soluble LRP6-ICD

The canonical Wnt/β-catenin (Wnt) pathway is a master transcriptional regulatory signaling pathway that controls numerous biological processes including proliferation and differentiation. As such, transcriptional activity of the Wnt pathway is tightly regulated and/or modulated by numerous proteins at the level of the membrane, cytosol and/or nucleus. In the nucleus, transcription of Wnt target genes by TCF/LEF-1 is repressed by the long Groucho/TLE co-repressor family. However, a truncated member of the Groucho/TLE family, amino terminal enhancer of Split (AES) can positively modulate TCF/LEF-1 activity by antagonizing long Groucho/TLE members in a dominant negative manner. We have previously shown the soluble intracellular domain of the LRP6 receptor, a receptor required for activation of the Wnt pathway, can positively regulate transcriptional activity within the Wnt pathway. In the current study, we show the soluble LRP6 intracellular domain (LRP6-ICD) can also translocate to the nucleus in CHO and HEK 293T cells and in contrast to cytosolic LRP6-ICD; nuclear LRP6-ICD represses TCF/LEF-1 activity. In agreement with previous reports, we show AES enhances TCF/LEF-1 mediated reporter transcription and further we demonstrate that AES activity is spatially regulated in HEK 293T cells. LRP6-ICD interacts with AES exclusively in the nucleus and represses AES mediated TCF/LEF-1 reporter transcription. These results suggest that LRP6-ICD can differentially modulate Wnt pathway transcriptional activity depending upon its subcellular localization and differential protein-protein interactions