Bacterial flora-typing with targeted, chip-based Pyrosequencing

<p>Abstract</p> <p>Background</p> <p>The metagenomic analysis of microbial communities holds the potential to improve our understanding of the role of microbes in clinical conditions. Recent, dramatic improvements in DNA sequencing throughput and cost will enable such analyses on individuals. However, such advances in throughput generally come at the cost of shorter read-lengths, limiting the discriminatory power of each read. In particular, classifying the microbial content of samples by sequencing the < 1,600 bp 16S rRNA gene will be affected by such limitations.</p> <p>Results</p> <p>We describe a method for identifying the phylogenetic content of bacterial samples using high-throughput Pyrosequencing targeted at the 16S rRNA gene. Our analysis is adapted to the shorter read-lengths of such technology and uses a database of 16S rDNA to determine the most specific phylogenetic classification for reads, resulting in a weighted phylogenetic tree characterizing the content of the sample. We present results for six samples obtained from the human vagina during pregnancy that corroborates previous studies using conventional techniques.</p> <p>Next, we analyze the power of our method to classify reads at each level of the phylogeny using simulation experiments. We assess the impacts of read-length and database completeness on our method, and predict how we do as technology improves and more bacteria are sequenced. Finally, we study the utility of targeting specific 16S variable regions and show that such an approach considerably improves results for certain types of microbial samples. Using simulation, our method can be used to determine the most informative variable region.</p> <p>Conclusion</p> <p>This study provides positive validation of the effectiveness of targeting 16S metagenomes using short-read sequencing technology. Our methodology allows us to infer the most specific assignment of the sequence reads within the phylogeny, and to identify the most discriminative variable region to target. The analysis of high-throughput Pyrosequencing on human flora samples will accelerate the study of the relationship between the microbial world and ourselves.</p

Antitumor effect of IP-10 by using two different approaches: Live delivery system and gene therapy

Purpose: Immunotherapy is one of the treatment strategies for breast cancer, the most common cancer in women worldwide. In this approach, the patient�s immune system is stimulated to attack microscopic tumors and control metastasis. Here, we used interferon γ-induced protein 10 (IP-10), which induces and strengthens antitumor immunity, as an immunotherapeutic agent. We employed Leishmania tarentolae, a nonpathogenic lizard parasite that lacks the ability to persist in mammalian macrophages, was used as a live delivery system for carrying the immunotherapeutic agent. It has been already shown that arginase activity, and consequently, polyamine production, are associated with tumor progression. Methods: A live delivery system was constructed by stable transfection of pLEXSY plasmid containing the IP-10-enhanced green fluorescent protein (IP-10- egfp) fusion gene into L. tarentolae. Then, the presence of the IP-10-egfp gene and the accurate integration location into the parasite genome were confirmed. The therapeutic efficacy of IP- 10 delivered via L. tarentolae and recombinant pcDNA-(IP- 10-egfp) plasmid was compared by determining the arginase activity in a mouse 4T1 breast cancer model. Results: The pcDNA- (IP-10-egfp) group showed a significant reduction in tumor weight and growth. Histological evaluation also revealed that only this group demonstrated inhibition of metastasis to the lung tissue. The arginase activity in the tissue of the pcDNA-(IP- 10-egfp) mice significantly decreased in comparison with that in normal mice. No significant difference was observed in arginase activity in the sera of mice receiving other therapeutic strategies. Conclusion: Our data indicates that IP-10 immunotherapy is a promising strategy for breast cancer treatment, as shown in the 4T1-implanted BALB/c mouse model. However, the L. tarentolae- (IP-10-EGFP) live delivery system requires dose modifications to achieve efficacy in the applied regimen (six injections in 3 weeks). Our results indicate that the arginase assay could be a good biomarker to differentiate tumoral tissues from the normal ones. © 2016 Korean Breast Cancer Society. All rights reserved

Green's functions and Hadamard parametrices for vector and tensor fields in general linear covariant gauges

We determine the retarded and advanced Green’s functions and Hadamard parametrices in curved spacetimes for linearized massive and massless gauge bosons and linearized Einstein gravity with a cosmological constant in general linear covariant gauges. These vector and tensor parametrices contain additional singular terms compared with their Feynman/de Donder-gauge counterpart. We also give explicit recursion relations for the Hadamard coefficients, and indicate their generalization to n dimensions. Furthermore, we express the divergence and trace of the vector and tensor Green’s functions in terms of derivatives of scalar and vector Green’s functions, and show how these relations appear as Ward identities in the free quantum theory

Association of anthropometric measures with cardiovascular risk factors and metabolic syndrome in normal-weight children and adolescents: The CASPIAN III study

Objective: This nationwide study was conducted to determine the association of anthropometric measures with cardiovascular risk factors and metabolic syndrome (MetS) in Iranian normal-weight children and adolescents. Methods: We analyzed the data of 3,565 children and adolescents (50.3 boys), aged 10-18 years, with a normal BMI (5th-84th percentile) obtained from the third survey of 'Childhood and Adolescence Surveillance and Prevention of Adult Non-communicable Disease' (CASPIAN III) study. The diagnostic criteria for MetS were defined by the International Diabetes Federation consensus. Results: The prevalence of MetS for 10- to 13.9-year-old boys, 14- to 18-year-old boys, 10- to 13.9-year-old girls, and 14- to 18-year-old girls were 1.4, 2.8, 2.3, and 3.3, respectively. After adjustment for age and sex, each unit increase in BMI (within normal range) and waist circumference increased the odds of MetS from 6 to 72 and from 1 to 20, respectively. The dominant pattern of dyslipidemia among the participants was high triglycerides and low high-density lipoprotein cholesterol. Conclusion: This study complements recent research about the high frequency of metabolic risk factors among normal-weight individuals in the pediatric age group. © 2013 S. Karger GmbH, Freiburg

Role of polymorphisms of the endothelial nitric oxide synthase gene in predicting slow-flow phenomenon after primary percutaneous coronary intervention

Objective: The aim of the present study was to examine the association between 2 polymorphisms of the endothelial nitric oxide (eNOS) gene (-786T>C and +894G>T) and the no-reflow/slow-flow phenomenon in post-primary percutaneous coronary intervention (PPCI) patients. Methods: A total of 103 post-PPCI patients were enrolled. Coronary no-reflow phenomenon was defined as a Thrombolysis in Myocardial Infarction (TIMI) flow grade 0-1 and coronary slow-flow phenomenon (CSFP) was defined as a TIMI flow grade �2. Results: Due to the small number of post-PPCI patients with the no-reflow phenomenon (n=4), the primary comparison was made between CSFP (n=20) and normal flow (n=83) groups. There was a greater frequency of CSFP among carriers of the-786C allele of the eNOS-786T>C polymorphism (odds ratio OR: 3.90; 95% confidence interval CI: 0.87-17.45; p=0.07). However, no such association was detected between the +894T allele of the eNOS +894G>T and CSFP (OR: 0.92; 95% CI: 0.21-3.98; p=0.91). In the adjusted analysis, the-786T>C polymorphism did not reach statistical significance. Conclusion: There was no significant association between CSFP and 2 of the most common polymorphisms of the eNOS gene in post-PPCI patients. © 2020 Turkish Society of Cardiology

DNA plasmid coding for Phlebotomus sergenti salivary protein PsSP9, a member of the SP15 family of proteins, protects against Leishmania tropica

Background: The vector-borne disease leishmaniasis is transmitted to humans by infected female sand flies, which transmits Leishmania parasites together with saliva during blood feeding. In Iran, cutaneous leishmaniasis (CL) is caused by Leishmania (L.) major and L. tropica, and their main vectors are Phlebotomus (Ph.) papatasi and Ph. sergenti, respectively. Previous studies have demonstrated that mice immunized with the salivary gland homogenate (SGH) of Ph. papatasi or subjected to bites from uninfected sand flies are protected against L. major infection. Methods and results: In this work we tested the immune response in BALB/c mice to 14 different plasmids coding for the most abundant salivary proteins of Ph. sergenti. The plasmid coding for the salivary protein PsSP9 induced a DTH response in the presence of a significant increase of IFN-γ expression in draining lymph nodes (dLN) as compared to control plasmid and no detectable PsSP9 antibody response. Animals immunized with whole Ph. sergenti SGH developed only a saliva-specific antibody response and no DTH response. Mice immunized with whole Ph. sergenti saliva and challenged intradermally with L. tropica plus Ph. sergenti SGH in their ears, exhibited no protective effect. In contrast, PsSP9-immunized mice showed protection against L. tropica infection resulting in a reduction in nodule size, disease burden and parasite burden compared to controls. Two months post infection, protection was associated with a significant increase in the ratio of IFN-γ to IL-5 expression in the dLN compared to controls. Conclusion: This study demonstrates that while immunity to the whole Ph. sergenti saliva does not induce a protective response against cutaneous leishmaniasis in BALB/c mice, PsSP9, a member of the PpSP15 family of Ph. sergenti salivary proteins, provides protection against L. tropica infection. These results suggest that this family of proteins in Ph. sergenti, Ph. duboscqi and Ph. papatasi may have similar immunogenic and protective properties against different Leishmania species. Indeed, this anti-saliva immunity may act as an adjuvant to accelerate the cell-mediated immune response to co-administered Leishmania antigens, or even cause the activation of infected macrophages to remove parasites more efficiently. These findings highlight the idea of applying arthropod saliva components in vaccination approaches for diseases caused by vector-borne pathogens. © 2019, Public Library of Science. All rights reserved

The effects of some antibiotics from cephalosporin groups on the acetylcholinesterase and butyrylcholinesterase enzymes activities in different tissues of rats

In our study, it was aimed to investigate the effects of cefazolin, cefuroxime, and cefoperazon injected to rats on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzyme activities in the heart, brain, eye, liver, and kidney tissues of rats. Liver AChE activity at the 1st and 3rd hours of cefuroxime groups was higher than the control group at the same time (p <.05). The AChE activity of the heart tissue decreased in the cefazolin group compared to the control group at the same hour, whereas it increased in the cefuroxime group (p <.05). AChE activities of kidney tissue of cefazolin and cefuroxime groups were lower than those of the same control group on the 3rd and started to increase on the next hours (p <.05). BChE activity is measured in tissues increased within the first three hours and decreased significantly within the first hour in the cefoperazone group (p <.05)

The effects of zingerone against vancomycin-induced lung, liver, kidney and testis toxicity in rats: The behavior of some metabolic enzymes

In this study, it was demonstrated the ameliorative effect of zingerone (ZO) (25 and 50 mg/kg body weight) against vancomycin (VCM) (200 mg/kg body weight) administered to rats on some metabolic enzymes’ activities in the lung, liver, kidney, and testis tissues of rats. Forty-two rats were divided into six groups as follows: control, ZO-25, ZO-50, VCM, VCM + ZO-25, and VCM + ZO-50. α-Glycosidase, butyrylcholinesterase, aldose reductase, acetylcholinesterase, paraoxonase-1, and carbonic anhydrase enzyme activities were significantly (P '.05) decreased in VCM group when compared with the control group. ZO, supplied with VCM, significantly activated some of these enzyme in all tissues. The results of this study showed that ZO regulates abnormal increases and decreases in VCM-induced metabolic enzyme activities in all tissues. © 2019 Wiley Periodicals, Inc

The Influence of Some Nonsteroidal Anti-inflammatory Drugs on Metabolic Enzymes of Aldose Reductase, Sorbitol Dehydrogenase, and α-Glycosidase: a Perspective for Metabolic Disorders

Pain, as a sensible alarm signal of living organisms to avoid tissue damage, is a common and debilitating consequence of a lot of disorders and diseases. The management of chronic pain is particularly challenging. For pain treatment, many analgesic drugs are used for their therapeutic effects. In this study, some nonsteroidal anti-inflammatory drugs including etofenamate, meloxicam, diclofenac, and tenoxicam were tested against α-glycosidase from Saccharomyces cerevisiae, sorbitol dehydrogenase (SDH), and aldose reductase (AR) enzymes from sheep liver. Nonsteroidal anti-inflammatory drugs demonstrated useful inhibition properties against α-glycosidase, AR, and SDH enzymes. Ki values were found in the range of 11.93 ± 3.77–364.88 ± 40.01 μM for α-glycosidase, 3.36 ± 1.08μM–17.68 ± 3.39 mM for AR, and 1.68 ± 0.02 μM–30.98 ± 14.31 mM for SDH. They can be selective drugs as antidiabetic agents, because of their inhibitory properties against SDH, α-glycosidase, and AR enzymes. © 2019, Springer Science+Business Media, LLC, part of Springer Nature