Biosynthesis,characterization and antimicrobial activity of silver nanoparticles by a halotolerant Bacillus endophyticus SCU-L

Abstract

We have conducted a thorough study on extracellular biosynthesis of silver nanoparticles (AgNPs) by a halotolerant bacterium Bacillus endophyticus SCU-L, which was identified by 16S rRNA gene sequencing analysis. This strain was selected during an ongoing research programme aimed at finding a novel biological method for green nanosynthetic routes using the extremophiles in unexplored hypersaline habitats. The biosynthesized AgNPs were characterized and analyzed with UV–vis spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, atomic force microscopy and X-ray diffraction. Further, the AgNPs were found to be spherical in shape with an average particle size of about 5.1?nm, and it was stable in aqueous solution for three months period of storage at room temperature under dark condition. Also, the synthesized AgNPs significantly presented antimicrobial activity against Candida albicans, Escherichia coli, Salmonella typhi and Staphylococcus aureus. The above results suggested that the present work may provide a valuable reference and theoretical basis for further exploration on microbial biosynthesis of AgNPs by halotolerant bacteria.     

马尾松幼苗根系和光合碳供应对土壤性质变化和温室气体排放的驱动作用

森林植被受全球气候变化、森林经营活动及病虫害等多种干扰,导致林地光合碳供应水平及根系输入量发生变化。在此背景下,土壤性质及土壤温室气体排放的响应及其机理是预测森林碳汇功能变化及森林可持续经营的重要依据。以2年生马尾松盆栽苗为对象,通过单株/盆和3株/盆栽植密度控制根系输入量、通过环割和截干控制光合碳向地下的供应能力,模拟森林植被干扰导致的根系输入量及光合碳供应变化对土壤理化性质、微生物群落结构及温室气体排放的影响。结果表明,苗木根系非结构性碳水化合物(TNC)含量和氮含量比单株/盆低;3株/盆的土壤速效氮含量比单株/盆低,土壤革兰氏阳性菌、厌氧菌、放线菌及丛枝菌根真菌丰富度均比单株/盆显著增加,3株/盆的土壤二氧化碳(CO₂)排放速率较高,但土壤氧化亚氮(N₂O)排放速率差异不显著。无论是单株/盆还是3株/盆,环割和截干处理后,根系生物量、根系长度及表面积均比对照显著下降;根系TNC含量显著下降。土壤和根系氮含量都有增加趋势;土壤微生物生物量碳(SMBC)含量降低,而土壤微生物生物量氮(SMBN)则提高。环割和截干后,土壤中各种微生物组成丰富度均有下降趋势,土壤CO₂排放速率显著下降,土壤N₂O排放速率则显著提高。根系输入量及光合碳供应对土壤细菌和真菌含量均有显著影响,土壤细菌含量与根系生物量、SMBC和SMBN显著正相关;土壤真菌含量与土壤温度显著负相关,与根系生物量、SMBC和SMBN显著正相关。相关分析表明,土壤CO₂排放通量与土壤温度、土壤湿度及根系生物量显著正相关,与土壤硝态氮显著负相关;土壤N₂O排放通量与土壤温度和土壤湿度显著正相关。以上研究表明,根系输入量与地上光合碳供应共同作用,改变土壤理化性质及微生物环境,进而影响土壤温室气体排放。     

Histamine releasing factor and elongation factor 1 alpha secreted via malaria parasites extracellular vesicles promote immune evasion by inhibiting specific T cell responses

Protozoan pathogens secrete nanosized particles called extracellular vesicles (EVs) to facilitate their survival and chronic infection. Here, we show the inhibition by Plasmodium berghei NK65 blood stage‐derived EVs of the proliferative response of CD4⁺ T cells in response to antigen presentation. Importantly, these results were confirmed in vivo by the capacity of EVs to diminish the ovalbumin‐specific delayed type hypersensitivity response. We identified two proteins associated with EVs, the histamine releasing factor (HRF) and the elongation factor 1α (EF‐1α) that were found to have immunosuppressive activities. Interestingly, in contrast to WT parasites, EVs from genetically HRF‐ and EF‐1α‐deficient parasites failed to inhibit T cell responses in vitro and in vivo. At the level of T cells, we demonstrated that EVs from WT parasites dephosphorylate key molecules (PLCγ1, Akt, and ERK) of the T cell receptor signalling cascade. Remarkably, immunisation with EF‐1α alone or in combination with HRF conferred a long‐lasting antiparasite protection and immune memory. In conclusion, we identified a new mechanism by which P. berghei‐derived EVs exert their immunosuppressive functions by altering T cell responses. The identification of two highly conserved immune suppressive factors offers new conceptual strategies to overcome EV‐mediated immune suppression in malaria‐infected individuals.     

Androgen receptor suppresses vasculogenic mimicry in hepatocellular carcinoma via circRNA7/miRNA7‐5p/VE‐cadherin/Notch4 signalling

Androgen receptor (AR) can suppress hepatocellular carcinoma (HCC) invasion and metastasis at an advanced stage. Vasculogenic mimicry (VM), a new vascularization pattern by which tumour tissues nourish themselves, is correlated with tumour progression and metastasis. Here, we investigated the effect of AR on the formation of VM and its mechanism in HCC. The results suggested that AR could down‐regulate circular RNA (circRNA) 7, up‐regulate micro RNA (miRNA) 7‐5p, and suppress the formation of VM in HCC Small hairpin circR7 (ShcircR7) could reverse the impact on VM and expression of VE‐cadherin and Notch4 increased by small interfering AR (shAR) in HCC, while inhibition of miR‐7‐5p blocked the formation of VM and expression of VE‐cadherin and Notch4 decreased by AR overexpression (oeAR) in HCC. Mechanism dissection demonstrated that AR could directly target the circR7 host gene promoter to suppress circR7, and miR‐7‐5p might directly target the VE‐cadherin and Notch4 3′UTR to suppress their expression in HCC. In addition, knockdown of Notch4 and/or VE‐cadherin revealed that shVE‐cadherin or shNotch4 alone could partially reverse the formation of HCC VM, while shVE‐cadherin and shNotch4 together could completely suppress the formation of HCC VM. Those results indicate that AR could suppress the formation of HCC VM by down‐regulating circRNA7/miRNA7‐5p/VE‐Cadherin/Notch4 signals in HCC, which will help in the design of novel therapies against HCC.     

A specific PCR assay for the diagnosis of Clonorchis sinensis infection in humans, cats and fishes

Clonorchiasis caused by Clonorchis sinensis is a fish-borne parasitic disease which is endemic in a number of countries. Using the sequences of the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) of C. sinensis as genetic markers, a pair of C. sinensis-specific primers was designed and used to establish a specific PCR assay for the diagnosis of C. sinensis infection in humans, cats and fish. This approach allowed the specific identification of C. sinensis after optimizing amplification conditions, with no amplicons being amplified from related heterogeneous DNA samples, and sequencing of amplicons confirmed the identity of the sequences amplified. The detection limit of this assay was 1.03 pg of adult C. sinensis, 1.1 metacercariae per gram of fish filet, and a single egg in human and cat feces. The PCR assay should provide a useful tool for the diagnosis and molecular epidemiological investigation of clonorchiasis in humans and animals.     

Effects of cotton cultivars on the population dynamics and population parameters of Tetranychus turkestani

土耳其斯坦叶螨Tetranychus turkestani(Vgorag et Nikolski)是新疆棉花害螨的优势种,本试验通过调查土耳其斯坦叶螨对棉田为害程度,研究了土耳其斯坦叶螨对新疆主栽的6个棉花品种(系)的种群动态进行了比较:新海21号、新陆早26号2个棉花品种上叶螨密度水平较低,标杂A1、297-5、81-3、新陆早12号叶螨密度没有显著差异.在此基础上比较了6种品种(系)上土耳其斯坦叶螨的实验种群参数:土耳其斯坦叶螨在新海21号、新陆早12号、新陆早26号品种上各虫态发育历期较长,各虫态存活率较低,每雌产卵量较少,种群趋势指数较低;而在标杂A1、297-5、81-3品种上的发育历期较短,各虫态存活率较高,每雌产卵量较多,种群趋势指数也较高.     

Human IgG Fc-glycosylation profiling reveals associations with age, sex, female sex hormones and thyroid cancer

IgG functions rely on interactions of the Fc region with other proteins, which are optimized by tailoring of a conserved N-linked glycosylation at Asn-297. We conducted a study involving 735 control individuals and 138 thyroid cancer patients. Here we demonstrated that previously described age-related change in Fc-glycosylation was further characterized by definite sex specificity. In females, the incidences of most of glycosylated forms began to pose characteristic changes at ages of puberty or menopause. In addition, glycan-glycan relationships existed extensively within Fc glycosylation, which were characterized to be altered upon different states of subjects, such as age, sex and thyroid cancer. In thyroid cancer patients, detailed comparison of glycosylation incidences with control individuals yielded insight into aberrant change in IgG(1) Fc-glycosylation. This aberrant pattern was also featured by remarkable specificities of both age and sex. The receiver operating characteristic curve analysis was used to determine diagnostic values of Fc glycosylation. Finally, clinical measurement of two major female sex hormones estradiol and progesterone was conducted to determine potential associations of hormones with IgG Fc glycosylation. This study provided an important view to the associations of IgG Fc N-linked glycosylation with age, sex, female sex hormones and thyroid cancer. This article is part of a Special Issue entitled: Proteomics: The clinical link.     

Functional analysis for gut microbes of the brown tree frog (Polypedates megacephalus) in artificial hibernation

Background

Annual hibernation is an adaptation that helps many animals conserve energy during food shortage in winter. This natural cycle is also accompanied by a remodeling of the intestinal immune system, which is an aspect of host biology that is both influenced by, and can itself influence, the microbiota. In amphibians, the bacteria in the intestinal tract show a drop in bacterial counts. The proportion of pathogenic bacteria is greater in hibernating frogs than that found in nonhibernating frogs. This suggests that some intestinal gut microbes in amphibians can be maintained and may contribute to the functions in this closed ecosystem during hibernation. However, these results were derived from culture-based approaches that only covered a small portion of bacteria in the intestinal tract.

Methods

In this study, we use a more comprehensive analysis, including bacterial appearance and functional prediction, to reveal the global changes in gut microbiota during artificial hibernation via high-throughput sequencing technology.

Results

Our results suggest that artificial hibernation in the brown tree frog (Polypedates megacephalus) could reduce microbial diversity, and artificially hibernating frogs tend to harbor core operational taxonomic units that are rarely distributed among nonhibernating frogs. In addition, artificial hibernation increased significantly the relative abundance of the red-leg syndrome-related pathogenic genus Citrobacter. Furthermore, functional predictions via PICRUSt and Tax4Fun suggested that artificial hibernation has effects on metabolism, disease, signal transduction, bacterial infection, and primary immunodeficiency.

Conclusions

We infer that artificial hibernation may impose potential effects on primary immunodeficiency and increase the risk of bacterial infections in the brown tree frog.

     

Prognostic Significance of Tumor Volume in Locally Recurrent Nasopharyngeal Carcinoma Treated with Salvage Intensity-Modulated Radiotherapy

Introduction

To evaluate the prognostic value of gross tumor volume (TV) in patients with locally recurrent, nonmetastatic nasopharyngeal carcinoma.

Methods

Between 2001 and 2012, 291 consecutive patients with locally recurrent, nonmetastatic nasopharyngeal carcinoma underwent salvage IMRT were retrospectively reviewed. The correlations between TV and recurrent T classification were analyzed. Survival analyses were performed. Receiver operating characteristic (ROC) curves were calculated to identify cut-off point of TV. The Akaike information criterion and Harrell’s concordance index (c-index) were utilized to test the prognostic value.

Results

The median TV significantly increased with advancing recurrent T classification (P<0.001). The 5-year overall survival rate was 33.2% for the entire cohort. On multivariate analysis, TV was an independent negative prognostic factor for distant metastasis-free survival (hazard ratio =1.013, P =0.003), overall survival (hazard ratio = 1.015, P<0.001) and toxicity-related death (hazard ratio = 1.014, P<0.001). The 5-year overall survival rates were 63.1% and 20.8% for patients with a TV < 22 cm³ and TV ≥22 cm³, respectively (P < 0.001). In patient with TV <22 cm³, locoregional failure is the leading cause of death. In patients with TV≥22 cm³, distant metastasis rate is higher and occurred within short term after local recurrence; meanwhile, radiation-induced injuries became more common and led to half of deaths in this group. The Akaike information criterion and c-index analyses indicated that the predictive ability of recurrent T classification improved when combined with TV.

Conclusions

Our data suggests TV is a significant prognostic factor for predicting the distant metastasis, overall survival and toxicity-related death of patients with locally recurrent, nonmetastatic nasopharyngeal carcinoma after salvage IMRT. TV should be considered when designing personalized salvage treatments for these patients. For patients with bulky local recurrent tumor, radiation may need to be de-emphasized in favor of systemic treatment or best supportive care.