Gastroenterologist perceptions of faecal microbiota transplantation

AIM: To explore gastroenterologist perceptions towards and experience with faecal microbiota transplantation(FMT).METHODS: A questionnaire survey consisting of 17 questions was created to assess gastroenterologists' attitude towards and experience with FMT. This was anonymously distributed in hard copy format amongst attendees at gastroenterology meetings in Australia between October 2013 and April 2014. Basic descriptive statistical analyses were performed.RESULTS: Fifty-two clinicians participated. Twenty one percent had previously referred patients for FMT,8% more than once. Ninety percent would refer patients with Clostridium difficile infection(CDI) for FMT if easily available,37% for ulcerative colitis,13% for Crohn's disease and 6% for irritable bowel syndrome. Six percent would not refer any indication,including recurrent CDI. Eighty-six percent would enroll patients in FMT clinical trials. Thirty-seven percent considered the optimal mode of FMT administration transcolonoscopic,17% nasoduodenal,13% enema and 8% oral capsule. The greatest concerns regarding FMT were: 42% lack of evidence,12% infection risk,10% non infectious adverse effects/lack of safety data,10% aesthetic,10% lack of efficacy,4% disease exacerbation,and 2% inappropriate use; 6% had no concerns. Seventy seven percent believed there is a lack of accessibility while 52% had an interest in learning how to provide FMT. Only 6% offered FMT at their institution.CONCLUSION: Despite general enthusiasm,most gastroenterologists have limited experience with,or access to,FMT. The greatest concerns were lack of supportive evidence and safety issues. However a significant proportion would refer indications other than CDI for FMT despite insufficient evidence. These data provide guidance on where education and training are required.

Oral Campylobacter species: initiators of a subgroup of inflammatory bowel disease?

In recent years,a number of studies detected a significantly higher prevalence of Campylobacter species such as Campylobacter concisus(C. concisus) in intestinal biopsies and fecal samples collected from patients with inflammatory bowel disease(IBD) compared to controls. Most of these Campylobacter species are not of zoonotic origin but are human oral Campylobacter species. Bacterial species usually cause diseases in the location where they colonize. However,C. concisus and other oral Campylobacter species are associated with IBD occurring at the lower parts of the gastrointestinal tract,suggesting that these Campylobacter species may have unique virulence factors that are expressed in the lower parts of the gastrointestinal tract.

变性梯度凝胶电泳(DGGE)在微生物生态学中的应用

由于从环境样品中分离和培养细菌的困难 ,分子生物学方法已发展用来描述和鉴定微生物群落。近年来基于 DNA方法的群落分析得到了迅速的发展 ,如 PCR扩增技术 ,克隆文库法 ,荧光原位杂交法 ,限制性酶切片段长度多态性法 ,变性和温度梯度凝胶电泳法。DGGE已广泛用于分析自然环境中细菌、蓝细菌 ,古菌、微微型真核生物、真核生物和病毒群落的生物多样性。这一技术能够提供群落中优势种类信息和同时分析多个样品。具有可重复和容易操作等特点 ,适合于调查种群的时空变化 ,并且可通过对切下的带进行序列分析或与特异性探针杂交分析鉴定群落成员。DGGE分析微生物群落的一般步骤如下 :一是核酸的提取 ,二是 1 6Sr RNA,1 8S r RNA或功能基因如可容性甲烷加单氧酶羟化酶基因 ( mmo X)和氨加单氧酶 α-亚单位基因 ( amo A)片段的扩增 ,三是通过DGGE分析 PCR产物。DGGE使用具有化学变性剂梯度的聚丙烯酰胺凝胶 ,该凝胶能够有区别的解链PCR扩增产物。由 PCR产生的不同的 DNA片段长度相同但核苷酸序列不同。因此不同的双链 DNA片段由于沿着化学梯度的不同解链行为将在凝胶的不同位置上停止迁移 [5] 。 DNA解链行为的不同导致一个凝胶带图案 ,该图案是微生物群落中主要种类的一个轮廓。 DGGE使用所有生物中保守的基因?

Developing More Effective Enhanced Biochar Fertilisers for Improvement of Pepper Yield and Quality

Utilization of biochar at high application rates can increase soil C and crop yields, decrease greenhouse gas emissions and reduce nutrient run-off from soils. However, the high application rate of 10 t ha-1may not return a profit to the farmer due to the high cost of biochar. In this study biochar was modified through pre-treating the biomass and post-treating with phosphoric acid, minerals and different chemical fertilisers to study the effects of two new enhanced biochar fertilisers on the yield and quality of green pepper in a field experiment with 5 fertilisation treatments and 3 replications. The two new biochar fertilisers significantly(P < 0.05) increased the yield of green pepper(11.33–11.47 t ha-1), compared with the conventional chemical fertiliser(9.72 t ha-1). The biochar fertiliser treatments improved the vitamin C content of green pepper from 236.99 to 278.28 mg kg-1, and also significantly(P < 0.05) reduced the nitrate content from 132.32 to 101.92 mg kg-1, compared with chemical fertiliser. This study indicated that, compared to the use of conventional chemical fertiliser, all of the biochar fertiliser treatments could significantly improve the yield and quality of green pepper.

Uptake and mitochondrial dysfunction of alpha-synuclein in human astrocytes, cortical neurons and fibroblasts

The accumulation and aggregation of alpha-synuclein(α-syn)in several tissue including the brain is a major pathological hallmark in Parkinson’s disease(PD).In this study,we show that α-syn can be taken up by primary human cortical neurons,astrocytes and skin-derived fibroblasts in vitro.Our findings that brain and peripheral cells exposed to α-syn can lead to impaired mitochondrial function,leading to cellular degeneration and cell death,provides additional evidence for the involvement of mitochondrial dysfunction as a mechanism of toxicity of α-syn in human cells.

Prolonged prothrombin time at admission predicts poor clinical outcome in COVID-19 patients

BACKGROUND Supported by the National Natural Science Foundation of China,No.The prognostic value of coagulation disorder in coronavirus disease 2019(COVID-19)patients should be demonstrated.AIM To investigate the abnormalities of coagulation parameters in the patients with COVID-19 and their prognostic values.METHODS Consecutive patients admitted in the isolation ward of Renmin Hospital of Wuhan University from January 31 to February 5,2020 with confirmed COVID-19 were included.The primary outcomes were death and survival as of March 11.Demographics,vital signs,comorbidities and laboratory tests were collected and compared between those who died and survivors.Logistic regression analysis for prognostic factors was performed.Kaplan-Meier analysis was used to compare the estimated survival rate between patients with prolonged prothrombin time and normal prothrombin time.RESULTS The total number of patients with confirmed COVID-19 who were enrolled was 213.The median age was 62 years,and 95 patients(44.6%)were men.Fifty-one patients were critical(23.9%),79 patients were severe(37.1%)and 83 patients were moderate(39%).As of March 11,2020,99 patients were discharged(46.5%),79 patients(37.1%)stayed in the hospital and 35 patients(16.2%)died.Median time to death was 6(4-8)d,while median hospital stay was 32(22-36)d in survivors(P<0.001).More men(P=0.002)and elderly patients(P<0.001)were found in the group of those who died.The respiration rate at admission was higher in the group of those who died(P<0.001).The incidences of hypertension(P=0.028),cerebrovascular disease(P<0.001),chronic kidney disease(P=0.02)and chronic obstructive pulmonary disease(P<0.001)were higher in the group of those who died.Platelet count was decreased in the group of those who died(P=0.002)whereas prothrombin time(P<0.001),activated partial thromboplastin time(P=0.033),concentration of D-dimer(P<0.001)and fibrin degradation products(P<0.001)were increased in the group of those who died.Prothrombin time[odds ratio(OR):2.19,P=0.004],respiration rate(OR:1.223,P<0.001),age(OR:1.074,P<0.001)and fibrin degradation products concentration(OR:1.02,P=0.014)were predictors of death.The survival rate was significantly lower in patients with prolonge CONCLUSION Prothrombin time,concentration of fibrin degradation products,respiration rate and age were predictive factors for clinical outcomes of COVID-19 patients.

Nonvesicular trafficking of cholesterol by Aster proteins

In a recent article published in Nature Metabolism,Peter Tontonoz and colleagues found that the Aster/GramD1 proteins were required for plasma membrane(PM)cholesterol to reach the endoplasmic reticulum(ER)in mouse liver during fasting,low-density lipoprotein(LDL)uptake,or reverse cholesterol transport(RCT).The Aster/GramD1 pathway plays a key role in maintaining hepatic and systemic cholesterol/lipid homeostasis.Cholesterol is an essential constituent of organellar membranes in mammalian cells[1].

The Role of RNA Structure in Posttranscriptional Regulation of Gene Expression

As more information is gathered on the mechanisms of transcription and translation, it is becoming apparent that these processes are highly regulated. The formation of mRNA secondary and tertiary structures is one such regulatory process that until recently it has not been analysed in depth. Formation of these mRNA structures has the potential to enhance and inhibit alternative splicing of transcripts, and regulate rates and amount of translation. As this regulatory mechanism potentially impacts at both the transcriptional and translational level, while also potentially utilising the vast array of non-coding RNAs, it warrants further investigation. Currently, a variety of high- throughput sequencing techniques including parallel analysis of RNA structure （PARS）, fragmentation sequencing （FragSeq） and selective 2-hydroxyl acylation analysed by primer extension （SHAPE） lead the way in the genome-wide identification and analysis of mRNA structure formation. These new sequencing techniques highlight the diversity and complexity of the transcriptome, and demonstrate another regulatory mechanism that could become a target for new therapeutic approaches.

PITPNC1 promotes the thermogenesis of brown adipose tissue under acute cold exposure

Brown adipose tissue (BAT) plays an essential role in non-shivering thermogenesis. The phosphatidylinositol transfer protein, cytoplasmic 1 (PITPNC1) is identified as a lipid transporter that reciprocally transfers phospholipids between intracellular membrane structures. However, the physiological significance of PITPNC1 and its regulatory mechanism remain unclear. Here, we demonstrate that PITPNC1 is a key player in thermogenesis of BAT. While Pitpnc1^(−/−) mice do not differ with wildtype mice in body weight and insulin sensitivity on either chow or high-fat diet, they develop hypothermia when subjected to acute cold exposure at 4℃. The Pitpnc1^(−/−) brown adipocytes exhibit defective β-oxidation and abnormal thermogenesis-related metabolism pathways in mitochondria. The deficiency of lipid mobilization in Pitpnc1^(−/−) brown adipocytes might be the result of excessive accumulation of phosphatidylcholine and a reduction of phosphatidic acid. Our findings have uncovered significant roles of PITPNC1 in mitochondrial phospholipid homeostasis and BAT thermogenesis.

Extended synaptotagmins, peroxisome-endoplasmic reticulum contact and cholesterol transport

Cholesterol is an essential component of mammalian cell membranes where it regulates membrane permeability,fluidity,fusion and cell signaling pathways (Chang et al.,2006).Cholesterol must be sorted precisely and transported efficiently for the proper functioning of membranes,as each cellular organelle has its distinct cholesterol concentration.The need to understand the molecular mechanisms governing cellular cholesterol transport and distribution is further highlighted by the link between abnormal subcellular distribution of cholesterol and a range of human diseases such as cancer (Zhuang et al.,2005),coronary heart disease (Zhang et al.,2008),steatohepatitis (Marí et al.,2006),neurodegenerative diseases including Alzheimer’s disease and Niemann Pick C (NP-C) disease (Vance et al.,2005),and early-onset obesity and insulin resistance (Liu et al.,2017).

Beta-and Novel Delta-Coronaviruses Are Identified from Wild Animals in the Qinghai-Tibetan Plateau, China

Outbreaks of severe virus infections with the potential to cause global pandemics are increasingly concerning. One type of those commonly emerging and re-emerging pathogens are coronaviruses(SARS-Co V, MERS-Co V and SARS-Co V-2).Wild animals are hosts of different coronaviruses with the potential risk of cross-species transmission. However, little is known about the reservoir and host of coronaviruses in wild animals in Qinghai Province, where has the greatest biodiversity among the world's high-altitude regions. Here, from the next-generation sequencing data, we obtained a known beta-coronavirus(beta-Co V) genome and a novel delta-coronavirus(delta-Co V) genome from faecal samples of 29 marmots, 50 rats and 25 birds in Yushu Tibetan Autonomous Prefecture, Qinghai Province, China in July 2019. According to the phylogenetic analysis, the beta-Co V shared high nucleotide identity with Coronavirus HKU24. Although the novel delta-Co V(Mt Co V) was closely related to Sparrow deltacoronavirus ISU42824, the protein spike of the novel delta-Co V showed highest amino acid identity to Sparrow coronavirus HKU17(73.1%). Interestingly, our results identified a novel host(Montifringilla taczanowskii) for the novel delta-Co V and the potential cross-species transmission. The most recent common ancestor(t MRCA) of Mt Co Vs along with other closest members of the species of Coronavirus HKU15 was estimated to be 289 years ago. Thus, this study increases our understanding of the genetic diversity of beta-Co Vs and deltaCo Vs, and also provides a new perspective of the coronavirus hosts.

Feeding Biochar to Cows:An Innovative Solution for Improving Soil Fertility and Farm Productivity

Addition of biochar produced through thermal decomposition of biomass has been seen as a strategy to improve soils and to sequester carbon(C), but wide scale implementation of the technology requires to devise innovative profitable solutions. To develop biochar utilisation with an integrated system approach, an innovative program was implemented in 2012 on a 53-ha farm in Western Australia to determine the costs and benefits of integrating biochar with animal husbandry and improvement of pastures. Biochar was mixed with molasses and fed directly to cows. The dung-biochar mixture was incorporated into the soil profile by dung beetles.We studied the changes in soil properties over 3 years. Biochar extracted from fresh dung and from the soil to a depth of 40 cm was characterised. A preliminary financial analysis of the costs and benefits of this integrated approach was also undertaken. The preliminary investigation results suggested that this strategy was effective in improving soil properties and increasing returns to the farmer. It was also concluded that the biochar adsorbed nutrients from the cow's gut and from the dung. Dung beetles could transport this nutrient-rich biochar into the soil profile. There was little evidence that the recalcitrant component of the biochar was reduced through reactions inside the gut or on/in the soil. Further research is required to quantify the long-term impact of integrating biochar and dung beetles into the rearing of cows.

Characterization of the archaeal community fouling a membrane bioreactor

Biofilm formation, one of the primary causes of biofouling, results in reduced membrane flux or increased transmembrane pressure and thus represents a major impediment to the wider implementation of membrane bioreactor （MBR） technologies for water purification. Most studies have focused on the role of bacteria in membrane fouling as they are the most dominant and best studied organisms present in the MBR. In contrast, there is limited information on the role of the archaeal community in biofilm formation in MBRs. This study investigated the composition of the archaeal community during the process of biofouling in an MBR. The archaeal community was observed to have lower richness and diversity in the biofilm than the sludge during the establishment ofbiofilms at low transmembrane pressure （TMP）. Clustering of the communities based on the Bray-Curtis similarity matrix indicated that a subset of the sludge archaeal community formed the initial biofilms. The archaeal community in the biofilm was mainly composed of Thermoprotei, Thermoplasmata, Thermococci, Methanopyri, Methanomicrobia and Halobacteria. Among them, the Thermoprotei and Thermoplasmata were present at higher relative proportions in the biofilms than they were in the sludge. Additionally, the Thermoprotei, Thermoplasmata and Thermococci were the dominant organisms detected in the initial biofilms at low TMP, while as the TMP increased, the Methanopyri, Methanomicrobia, Aciduliprofundum and Halobacteria were present at higher abundances in the biofilms at high TMP.

Sterol-binding proteins and endosomal cholesterol transport

Endosomal compartments sort and deliver exogenous lipoprotein-derived cholesterol to the endoplasmic reticulum for regulating cellular cholesterol homeostasis.A large number of studies have focused on the removal of endosomal cholesterol,since its accumulation leads to devastating human diseases.Recent studies suggest that cytoplasmic sterol-binding proteins may be involved in endosomal cholesterol transport.In particular,endosome/lysosome-localized or-associated cholesterol-binding proteins may serve as key mediators of cholesterol removal in a non-vesicular manner.Further characterization of these cholesterol-binding proteins will shed light on the molecular mechanisms that regulate endosomal cholesterol sorting.

A Lipidomic Perspective on Intermediates in Cholesterol Synthesis as Indicators of Disease Status

Lipidomics is increasingly becoming a viable method for researchers to routinely identify the various sterols present in samples, beyond just measuring cholesterol itself. In particular, the measurement of intermediates in cholesterol synthesis can shed new insights into not only the flux through the pathway, but also numerous disease states where levels of sterol intermediates are drastically altered. In this review, we indicate several intermediates that are relevant to disease, and discuss the challenges for analysing them, including the need for standardised methodology or universal controls across the lipidomics field.

Eye contact and sociability data suggests that Australian dingoes were never domesticated

Dogs were the first animal to become domesticated by humans,and they represent a classic model system for unraveling the processes of domestication.We compare Australian dingo eye contact and socialization with Basenji and German Shepherd dog(GSD)breeds.Australian dingoes arrived in Australia 5,000–8,000 BP,and there is debate whether they were domesticated before their arrival.The Basenji represents a primitive breed that diverged from the remaining breeds early in the domestication process,while GSDs are a breed dog selected from existing domestic dogs in the late 1800s.We conducted a 4-phase study with unfamiliar and familiar investigators either sitting passively or actively calling each canid.We found 75%of dingoes made eye contact in each phase.In contrast,86%of Basenjis and 96%of GSDs made eye contact.Dingoes also exhibited shorter eye-gaze duration than breed dogs and did not respond to their name being called actively.Sociability,quantified as a canid coming within 1m of the experimenter,was lowest for dingoes and highest for GSDs.For sociability duration,dingoes spent less time within 1m of the experimenter than either breed dog.When compared with previous studies,these data show that the dingo is behaviorally intermediate between wild wolves and Basenji dogs and suggest that it was not domesticated before it arrived in Australia.However,it remains possible that the accumulation of mutations since colonization has obscured historical behaviors,and dingoes now exist in a feralized retamed cycle.Additional morphological and genetic data are required to resolve this conundrum.

小鼠CDP-二酰基甘油合成酶-2(Cds2)肝脏特异性敲除导致线粒体功能受损并快速发展为非酒精性肝炎及肝纤维化

非酒精性脂肪性肝病(NAFLD)包括单纯脂肪浸润和非酒精性脂肪性肝炎(NASH),继续发展可导致更为严重的肝纤维化、肝硬化和肝癌等.诱导肝脏从单纯脂肪浸润发展成NASH的机制还不是特别清楚.本研究构建了肝脏特异敲除CDP-二酰基甘油合成酶-2(Cds2)小鼠,并发现该小鼠快速产生肝脏脂肪大量储积、肝炎和肝纤维化等一系列肝脏病变.CDS2定位在线粒体相关内质网膜(MAM)上,敲除Cds2会导致线粒体功能受损和线粒体磷脂酰乙醇胺含量下降.肝脏过表达线粒体磷脂酰乙醇胺合成酶(Pisd)会增加线粒体磷脂酰乙醇胺含量,并减轻肝脏Cds2敲除小鼠肝损伤的表型.另外,过表达Cds2可以抵抗高脂喂食诱导的脂肪肝和肥胖.本研究揭示了小鼠肝脏Cds2生理功能,即Cds2可以调节线粒体磷脂含量和线粒体功能并继而影响NASH的快速发生.

VPOT:A Customizable Variant Prioritization Ordering Tool for Annotated Variants

Next-generation sequencing(NGS) technologies generate thousands to millions of genetic variants per sample.Identification of potential disease-causal variants is labor intensive as it relies on filtering using various annotation metrics and consideration of multiple pathogenicity prediction scores.We have developed VPOT(variant prioritization ordering tool),a python-based command line tool that allows researchers to create a single fully customizable pathogenicity ranking score from any number of annotation values,each with a user-defined weighting.The use of VPOT can be informative when analyzing entire cohorts,as variants in a cohort can be prioritized.VPOT also provides additional functions to allow variant filtering based on a candidate gene list or by affected status in a family pedigree.VPOT outperforms similar tools in terms of efficacy,flexibility,scalability,and computational performance.VPOT is freely available for public use at Git Hub(https://github.com/VCCRI/VPOT/).Documentation for installation along with a user tutorial,a default parameter file,and test data are provided.

Drug targets beyond HMG-CoA reductase:Why venture beyond the statins?

In this review,we aim to convey a brief,select history of the development of cholesterol-lowering therapies.We focus particularly on the highly successful statins as well as setbacks that should serve as cautionary tales.We go on to preview recent developments that may complement,if not one day replace,the statins.Our focus is on pharmacological interventions,particularly those targeting the cholesterol biosynthetic pathway.Also,we examine therapies under current investigation that target the assembly of atherogenic lipoproteins(via apolipoprotein B or microsomal triglyceride transfer protein),the stability of the low-density lipoprotein-receptor(via PCSK9,proprotein convertase subtilisin kexin 9),or are designed to increase high-density lipoprotein-cholesterol(via inhibition of cholesteryl ester transfer protein).