Determination of plasma microRNA for early detection of gastric cancer

Gastric cancer is the fourth most prevalent malignancy worldwide and remains the second most common cause of cancer-related death globally. Understanding the molecular structure of gastric carcinogenesis might identify new diagnostic and therapeutic strategies for this disease. Thus, early detection of gastric cancer is a key measure to reduce the mortality and improve the prognosis of gastric cancer. There have recently been several reports that microRNAs (miRNAs) circulate in highly stable, cell-free forms in blood. Because serum and plasma miRNAs are relatively easy to access, circulating miRNAs also have great potential to serve as non-invasive biomarkers. Although a number of miRNAs associated with gastric cancer have been identified, the underlying mechanism of these miRNAs in tumorigenesis and tumor progression remains to be investigated. The purpose of this study is to identify the potential of serum miRNAs as biomarkers for early detection of gastric cancer patients. RNA was isolated using the High Pure miRNA Isolation Kit (Roche) following the manufacturer’s protocol. cDNA and preamplification protocols were obtained from the isolated plasma miRNAs. The BioMark? 96.96 Dynamic Array (Fluidigm Corporation) for real-time qPCR was used to simultaneously quantite the expression of 740 miRNAs. All statistical analyses were performed using the Biogazelle’s qbase PLUS 2.0 software. In this study, among 740 miRNAs that we analyzed only miR-195-5p was significantly (p < 0.05, fold changes = 13, 3) down-regulated in gastric cancer patients compared with control. We demonstrated that miR-195-5p is a novel tumor suppressor miRNA and may contribute to gastric carcinogenesis. The miRNA expression profile described in this study should contribute to future studies on the role of miRNAs in gastric cancer.     

A review of Turkish saproxylic beetles from the European Red List

The conservation of beetles that depend on dying or dead wood (i.e. saproxylic), has received a great deal of attention in many parts of the world in recent years. Human activities such as urbanization and logging, and their results, e.g. global warming, destroy natural ecosystems and threaten unusual species such as Lucanus cervus (L. 1758), Rosalia alpina (L. 1758), Limoniscus violaceus (Müller 1821) and Osmoderma eremita (Scopoli 1763). Attempts have been made to conserve the habitats upon which these species depend. Although represented by a large number of species, saproxylic beetles in Turkey have not received much attention. Although some Turkish studies have acknowledged their value, comprehensive investigations of these species are very limited. The aim of this study is to make a list, from the literature, of the species that are endangered in Europe and found in Turkey, to highlight the importance of these species. The research includes 151 saproxylic beetle species belonging to the families Bostrichidae, Cerambycidae, Elateridae, Eucnemidae, Scarabaeidae, Euchiridae, Lucanidae and Mycetophagidae. The list of Turkish saproxylic beetles was prepared using the European Red List of Saproxylic Beetles. Information is provided about the IUCN Red List category in Europe, the locations of the species in Turkey, and their global geographic range. We gathered all data about these species from the international literature. Furthermore, we also include in the list certain species whose locations are not specified, but which inhabit Turkey. We obtained their records from some catalogs in Turkey. In addition to the list, we prepared three distribution maps for Turkey. The maps show distribution in Turkey of some species in the Endangered, Vulnerable and Near Threatened categories in the European Red List. It is expected that this study will lead to comprehensive studies on saproxylic beetles from Turkey.     

The population genomics of multiple tsetse fly (Glossina fuscipes fuscipes) admixture zones in Uganda

Understanding the mechanisms that enforce, maintain or reverse the process of speciation is an important challenge in evolutionary biology. This study investigates the patterns of divergence and discusses the processes that form and maintain divergent lineages of the tsetse fly Glossina fuscipes fuscipes in Uganda. We sampled 251 flies from 18 sites spanning known genetic lineages and the four admixture zones between them. We apply population genomics, hybrid zone and approximate Bayesian computation to the analysis of three types of genetic markers: 55,267 double‐digest restriction site‐associated DNA (ddRAD) SNPs to assess genome‐wide admixture, 16 microsatellites to provide continuity with published data and accurate biogeographic modelling, and a 491‐bp fragment of mitochondrial cytochrome oxidase I and II to infer maternal inheritance patterns. Admixture zones correspond with regions impacted by the reorganization of Uganda's river networks that occurred during the formation of the West African Rift system over the last several hundred thousand years. Because tsetse fly population distributions are defined by rivers, admixture zones likely represent both old and new regions of secondary contact. Our results indicate that older hybrid zones contain mostly parental types, while younger zones contain variable hybrid types resulting from multiple generations of interbreeding. These findings suggest that reproductive barriers are nearly complete in the older admixture zones, while nearly absent in the younger admixture zones. Findings are consistent with predictions of hybrid zone theory: Populations in zones of secondary contact transition rapidly from early to late stages of speciation or collapse all together.     

Development of Vaccine Prototype Against Zika Virus Disease of Peptide-Loaded PLGA Nanoparticles and Evaluation of Cytotoxicity

Zika virus has recently evolved from an obscure mosquito-borne pathogen to an international public health concern. People with Zika virus disease can have indications including mild fever, skin rash, conjunctivitis, muscle pain, malaise or headache. Effective vaccines are needed for controlling and preventing the disease. In the current study, we aim to design the substructure for vaccine against Zika virus by forming antigenic peptide epitope of the disease. Zika peptide loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles have been fabricated in the present work as a potential artificial vaccine. UV and FT-IR Spectrophotometers and ZetaSizer were used for studying the nanoparticles, and Scanning Electron Microscope was used for morphological examination. The nanoparticles (NPs) yield, encapsulation efficiency, the peptide loading capacity were determined and in vitro release of the peptide was studied. Cytotoxic effects of the various concentrations of Zika peptide, blank PLGA nanoparticles and Zika peptide loaded PLGA nanoparticles on ECV304 human epithelial cells were determined via MTT assay. The present paper could be considered as one of the important steps in the use of nanoparticles for constructing the new generation of vaccination systems.

     

Trypanosome Infection Establishment in the Tsetse Fly Gut Is Influenced by Microbiome-Regulated Host Immune Barriers

Tsetse flies (Glossina spp.) vector pathogenic African trypanosomes, which cause sleeping sickness in humans and nagana in domesticated animals. Additionally, tsetse harbors 3 maternally transmitted endosymbiotic bacteria that modulate their host''s physiology. Tsetse is highly resistant to infection with trypanosomes, and this phenotype depends on multiple physiological factors at the time of challenge. These factors include host age, density of maternally-derived trypanolytic effector molecules present in the gut, and symbiont status during development. In this study, we investigated the molecular mechanisms that result in tsetse''s resistance to trypanosomes. We found that following parasite challenge, young susceptible tsetse present a highly attenuated immune response. In contrast, mature refractory flies express higher levels of genes associated with humoral (attacin and pgrp-lb) and epithelial (inducible nitric oxide synthase and dual oxidase) immunity. Additionally, we discovered that tsetse must harbor its endogenous microbiome during intrauterine larval development in order to present a parasite refractory phenotype during adulthood. Interestingly, mature aposymbiotic flies (Gmm ^Apo) present a strong immune response earlier in the infection process than do WT flies that harbor symbiotic bacteria throughout their entire lifecycle. However, this early response fails to confer significant resistance to trypanosomes. Gmm ^Apo adults present a structurally compromised peritrophic matrix (PM), which lines the fly midgut and serves as a physical barrier that separates luminal contents from immune responsive epithelial cells. We propose that the early immune response we observe in Gmm ^Apo flies following parasite challenge results from the premature exposure of gut epithelia to parasite-derived immunogens in the absence of a robust PM. Thus, tsetse''s PM appears to regulate the timing of host immune induction following parasite challenge. Our results document a novel finding, which is the existence of a positive correlation between tsetse''s larval microbiome and the integrity of the emerging adult PM gut immune barrier.     

CARD9-Dependent Neutrophil Recruitment Protects against Fungal Invasion of the Central Nervous System

Candida is the most common human fungal pathogen and causes systemic infections that require neutrophils for effective host defense. Humans deficient in the C-type lectin pathway adaptor protein CARD9 develop spontaneous fungal disease that targets the central nervous system (CNS). However, how CARD9 promotes protective antifungal immunity in the CNS remains unclear. Here, we show that a patient with CARD9 deficiency had impaired neutrophil accumulation and induction of neutrophil-recruiting CXC chemokines in the cerebrospinal fluid despite uncontrolled CNS Candida infection. We phenocopied the human susceptibility in Card9 ^-/- mice, which develop uncontrolled brain candidiasis with diminished neutrophil accumulation. The induction of neutrophil-recruiting CXC chemokines is significantly impaired in infected Card9 ^-/- brains, from both myeloid and resident glial cellular sources, whereas cell-intrinsic neutrophil chemotaxis is Card9-independent. Taken together, our data highlight the critical role of CARD9-dependent neutrophil trafficking into the CNS and provide novel insight into the CNS fungal susceptibility of CARD9-deficient humans.     

Optimization of pulsed field gel electrophoresis (PFGE) conditions for thermophilic <Emphasis Type="Italic">bacilli</Emphasis>

Although thermophilic members of the genus Bacillus are known to tolerate extreme environmental conditions, they appeared to be readily damaged upon mechanical manipulations. This fact was evidenced in genotyping of some strains of thermophilic Bacillus by pulsed field gel electrophoresis (PFGE). Consequently, a new procedure for the preparation of agarose plugs was developed. The procedure produced interpretable genomic DNA restriction profiles.     

Aflatoxin M<Subscript>1</Subscript> in human breast milk in southeastern Turkey

This study was performed to determine aflatoxin M₁ (AFM₁) in human breast milk samples collected in ?anl?urfa, located in Southeastern region of Turkey, and to investigate a possible correlation between AFM₁ occurrence (frequency and levels) and sampling seasons. Human breast milk samples collected in December 2014 and in June 2015 from a total of 74 nursing women, both outpatient and inpatient volunteers in hospitals located in ?anl?urfa, Turkey, were analyzed using competitive enzyme-linked immunosorbent assay (ELISA) for the presence of AFM₁. AFM₁ was detected in 66 (89.2%) out of 74 samples at an average concentration of 19.0 ± 13.0 ng/l (min.-max., 9.6–80 ng/l). There was a statistically significant difference between December and June concerning AFM₁ levels (p < 0.05). Further detailed studies will be needed to determine the main sources of aflatoxins in food, to establish protection strategies against maternal and infant exposure to these mycotoxins.