Piwi like RNA-mediated gene silencing 1 gene as a possible major player in gastric cancer

AIM To establish a permanent piwi like RNA-mediated genesilencing 1(PIWIL1) gene knockout in AGP01 gastric cancer cell line using CRISPR-Cas9 system and analyze phenotypic modifications as well as gene expression alterations.METHODS CRISPR-Cas9 system used was purchased from Dharmacon GE Life Sciences(Lafayette, CO, United States) and permanent knockout was performed according to manufacturer's recommendations. Woundhealing assay was performed to investigate the effect of PIWIL1 knockout on migration capability of cells and Boyden chamber invasion assay was performed to investigate the effect on invasion capability. For the gene expression analysis, a one-color microarray-based gene expression analysis kit(Agilent Technologies, Santa Clara, CA, United States) was used according to the protocol provided by the manufacturer. RESULTS PIWIL1 gene knockout caused a significant decrease in AGP01 migration capacity as well as a significant decrease in cell invasiveness. Moreover, functional analysis based on grouping of all differentially expressed m RNAs identified a total of 35 genes(5 up-regulated and 30 down-regulated) encoding proteins involved in cellular invasion and migration. According to current literature, 9 of these 35 genes(DOCK2, ZNF503, PDE4 D, ABL1, ABL2, LPAR1, SMAD2, WASF3 and DACH1) are possibly related to the mechanisms used by PIWIL1 to promote carcinogenic effects related to migration and invasion, since their functions are consistent with the changes observed(being up-or down-regulated after knockout). CONCLUSION Taken together, these data reinforce the idea that PIWIL1 plays a crucial role in the signaling pathway of gastric cancer, regulating several genes involved in migration and invasion processes; therefore, its use as a therapeutic target may generate promising results in the treatment of gastric cancer.

Calculation of Dose in Healthy Organs,during Radiotherapy 4-Field Box 3D Conformal for Prostate Cancer,Simulation of the Linac 2300,Radiotherapy Room and MAX Phantom

In men, prostate cancer is one of the most frequent types, and radiotherapy is adopted as a form of treatment. Although there are efforts to minimize the dose in the healthy organ and tissues adjacent to the tumor during radiotherapy, these organs are affected by the secondary scattered and leakage radiation originating from the therapeutic beam and these doses deposited in the healthy organs, can induce the appearance of new focal points of cancer. The aim of this study is to calculate the equivalent and effective doses, due to photons and neutrons, in healthy organs of a patient submitted to radiotherapy treatment for prostate cancer. Computed simulation of radiotherapy treatment for prostate cancer was used to perform the dose calculations, adopting the treatment protocol used at INCA (Brazilian National Cancer Institute). The MCNPX code was employed in the simulation radiation transport while the male voxel MAX phantom was used to represent the patient's human anatomy. The results obtained in this study indicate that the organs close to the irradiated region are predominantly affected by the dose due to photons, with an impact on organs from different systems of the body, such as the bladder, colon, and testicles, besides bone structures such as the femur, pelvis and spinal column. The results obtained from the doses deposited due to neutrons suggest that tibia and fibula, mandible, cranium, brain and thyroid, had the highest dose deposited due to neutrons in relation to photons. The result obtained from the effective dose was 31.47 mSv due to photons, while the dose due to neutrons was 0.42 mSv. Note that the effective dose due to photons is significantly higher than the effective dose due to neutrons. The values calculated in this study were compared with the experimental values obtained in the literature, presenting reasonable concordance. Additionally, as described in the literature, it was verified that the dose due to photons decreases considerably with the increase in the distance of the target organ, while the dose due to neutrons is distributed homogeneously in the organs. It is concluded that the contribution of neutrons to the appearance of secondary cancers is more relevant in the organs furthest from the target volume, and that organs close to the tumor, are affected predominantly by the dose due to photons.

Diet and microbiota in inflammatory bowel disease: The gut in disharmony

Bacterial colonization of the gut shapes both the local and the systemic immune response and is implicated in the modulation of immunity in both healthy and disease states. Recently, quantitative and qualitative changes in the composition of the gut microbiota have been detected in Crohn's disease and ulcerative colitis, reinforcing the hypothesis of dysbiosis as a relevant mechanism underlying inflammatory bowel disease(IBD) pathogenesis. Humans and microbes have coexisted and co-evolved for a long time in a mutually beneficial symbiotic association essential for maintaining homeostasis. However, the microbiome is dynamic, changing with age and in response to environmental modifications. Among such environmental factors, food and alimentary habits, progressively altered in modern societies, appear to be critical modulators of the microbiota, contributing to or co-participating in dysbiosis. In addition, food constituents such as micronutrients are important regulators of mucosal immunity, with direct or indirect effects on the gut microbiota. Moreover, food constituents have recently been shown to modulate epigenetic mechanisms, which can result in increased risk for the development and progression of IBD. Therefore, it is likely that a better understanding of the role of different food components in intestinal homeostasis and the resident microbiota will be essential for unravelling the complex molecular basis of the epigenetic, genetic and environment interactions underlying IBD pathogenesis as well as for offering dietary interventions with minimal side effects.

Thiopurine-methyltransferase variants in inflammatory bowel disease:Prevalence and toxicity in Brazilian patients

AIM: To analyze the prevalence of thiopurine-methyltransferase (TPMT) genotypes and their association with drug toxicity in inflammatory bowel disease (IBD) patients from southeastern Brazil.

Nicotinic cholinergic receptors in esophagus:Early alteration during carcinogenesis and prognostic value

AIM: To compare expression of nicotinic cholinergic receptors(CHRNs) in healthy and squamous cell carcinoma-affected esophagus and determine the prognostic value.METHODS: We performed RT-q PCR to measure the expression of CHRNs in 44 esophageal samples from healthy individuals and in matched normal surrounding mucosa, and in tumors from 28 patientsdiagnosed with esophageal squamous cell carcinoma(ESCC). Next, we performed correlation analysis for the detected expression of these receptors with the habits and clinico-pathological characteristics of all study participants. In order to investigate the possible correlations between the expression of the different CHRN subunits in both healthy esophagus and tissues from ESCC patients, correlation matrices were generated. Subsequently, we evaluated whether the detected alterations in expression of the various CHRNs could precede histopathological modifications during the esophageal carcinogenic processes by using receiver operating characteristic curve analysis. Finally, we evaluated the impact of CHRNA5 and CHRNA7 expression on overall survival by using multivariate analysis.RESULTS: CHRNA3, CHRNA5, CHRNA7 and CHRNB4, but not CHRNA1, CHRNA4, CHRNA9 or CHRNA10, were found to be expressed in normal(healthy) esophageal mucosa. In ESCC, CHRNA5 and CHRNA7 were overexpressed as compared with patient-matched surrounding non-tumor mucosa(ESCC-adjacent mucosa; P < 0.0001 and P = 0.0091, respectively). Positive correlations were observed between CHRNA3 and CHRNB4 expression in all samples analyzed. Additionally, CHRNB4 was found to be differentially expressed in the healthy esophagus and the normalappearing ESCC-adjacent mucosa, allowing for distinguishment between these tissues with a sensitivity of 75.86% and a specificity of 78.95%(P = 0.0002). Finally, CHRNA5 expression was identified as an independent prognostic factor in ESCC; patients with high CHRNA5 expression showed an increased overall survival, in comparison with those with low expression. The corresponding age- and tumor stage-adjusted hazard ratio was 0.2684(95%CI: 0.075-0.97, P = 0.0448).CONCLUSION: Expression of CHRNs is homogeneous along healthy esophagus and deregulated in ESCC, suggesting a pathogenic role for these receptors in ESCC development and progression.

Hepatitis B virus lineages in mammalian hosts:Potential for bidirectional cross-species transmission

The hepatitis B virus(HBV)is a cosmopolitan infectious agent currently affecting over 350 million people worldwide,presently accounting for more than two billion infections.In addition to man,other hepatitis virus strains infect species of several mammalian families of the Primates,Rodentia and Chiroptera orders,in addition to birds.The mounting evidence of HBV infection in African,Asian and neotropical primates draws attention to the potential crossspecies,zoonotic transmission of these viruses to man.Moreover,recent evidence also suggests the humans may also function as a source of viral infection to other mammals,particularly to domestic animals like poultry and swine.In this review,we list all evidence of HBV and HBVlike infection of nonhuman mammals and discuss their potential roles as donors or recipients of these viruses to humans and to other closely-related species.

Trichomonas vaginalis perturbs the junctional complex in epithelial cells

Trichomonas vaginalis,a protist parasite of the urogenital tract in humans,is the causative agent of trichomonosis,which in recent years have been associated with the cervical cancer development.In the present study we analyzed the modifications at the junctional complex level of Caco-2 cells after interaction with two isolates of T.vaginalis and the influence of the iron concentration present in the parasite’s culture medium on the interaction effects.Our results show that T.vaginalis adheres to the epithelial cell causing alterations in the junctional complex,such as:(a)a decrease in transepithelial electrical resistance;(b)alteration in the pattern of junctional complex proteins distribution as observed for E-cadherin,occludin and ZO-1;and(c)enlargement of the spaces between epithelial cells.These effects were dependent on(a)the degree of the parasite virulence isolate,(b)the iron concentration in the culture medium,and(c)the expression of adhesin proteins on the parasite surface.

An international survey of classification and treatment choices for group D retinoblastoma

AIM： To determine which IIRC scheme was used by retinoblastoma centers worldwide and the percentage of D eyes treated primarily with enucleation versus globe salvaging therapies as well as to correlate trends in treatment choice to IIRC version used and geographic region. METHODS： An anonymized electronic survey was offered to 115 physicians at 39 retinoblastoma centers worldwide asking about IIRC classification schemes and treatment patterns used between 2008 and 2012. Participants were asked to record which version of the IIRC was used for classification, how many group D eyes were diagnosed, and how many eyes were treated with enucleation versus globe salvaging therapies. Averages of eyes per treatment modality were calculated and stratified by both IIRC version and geographic region. Statistical significance was determined by Chi-square, ANOVA and Kruskal-Wallis tests using Prism. RESULTS： The survey was completed by 29% of physicians invited to participate. Totally 1807 D eyes were diagnosed. Regarding IIRC system, 27% of centers used the Children＇s Hospital of Los Angeles （CHLA） version, 33% used the Children＇s Oncology Group （COG） version, 23% used the Philadelphia version, and 17% were unsure. The rate for primary enucleation varied between 0 and 100% and the mean was 29%. By IIRC version, primary enucleation rates were： Philadelphia, 8%; COG, 34%; and CHLA, 37%. By geographic region, primary enucleation rates were： Latin America, 57%; Asia, 40%; Europe, 36%; Africa, 10%, US, 8%; and Middle East, 8%. However, systemic chemoreduction was used more often than enucleation in all regions except Latin America with a mean of 57% per center （P〈0.0001）. CONCLUSION： Worldwide there is no consensus on which IIRC version is used, systemic chemoreduction was the most frequently used initial treatment during the study period followed by enucleation and primary treatment modality, especially enucleation, varied greatly with regards to IIRC version used and geographic region.

Neurophysics Assessment of the Muscle Bioenergy Generated by Transcranial Magnetic Stimulation

Te content of the rectifed motor evoked potential(MEP)induced by transcranial magnetic stimulation(TMS)has ambiguously been assessed without the precision that energy calculation deserves.Tis fact has misled data interpretation and misguided biomedical interventions.To defnitively fll the gap that exits in the neurophysics processing of these signals,we computed,in Walls(̂�),the bioenergy within the rectifed MEP recorded from the human frst digitorum index(FDI)muscle at rest and under isometric contraction.We also gauged the biowork exerted by this muscle.Here we show that bioenergy and biowork can accurately and successfully be assessed,validated,and determined in̂�from MEP signals induced by TMS,regardless of knowing the mathematical expression of the function of the signal.Our novel neurophysics approach represents a dramatic paradigm shif in analysis and interpretation of the content of the MEP and will give a true meaning to the content of rectifed signals.Importantly,this innovative approach allowed unveiling that women exerted more bioenergy than men at the magnetic stimulations used in this study.Revisitation of conclusions drawn from studies published elsewhere assessing rectifed EMG signals that have used ambiguous units is strongly recommended.