Evaluation of the MODS culture technique for the diagnosis of tuberculous meningitis

Background

Tuberculous meningitis (TBM) is a devastating condition. The rapid instigation of appropraite chemotherapy is vital to reduce morbidity and mortality. However rapid diagnosis remains elusive; smear microscopy has extremely low sensitivity on cerebrospinal fluid (CSF) in most laboratories and PCR requires expertise with advanced infrastructure and has sensitivity of only around 60% under optimal conditions. Neither technique allows for the microbiological isolation of M. tuberculosis and subsequent drug susceptibility testing. We evaluated the recently developed microscopic observation drug susceptibility (MODS) assay format for speed and accuracy in diagnosing TBM.

Methodology/Principal Findings

Two hundred and thirty consecutive CSF samples collected from 156 patients clinically suspected of TBM on presentation at a tertiary referal hospital in Vietnam were enrolled into the study over a five month period and tested by Ziehl-Neelsen (ZN) smear, MODS, Mycobacterial growth Indicator tube (MGIT) and Lowenstein-Jensen (LJ) culture. Sixty-one samples were from patients already on TB therapy for >1day and 19 samples were excluded due to untraceable patient records. One hundred and fifty samples from 137 newly presenting patients remained. Forty-two percent (n = 57/137) of patients were deemed to have TBM by clinical diagnostic and microbiological criteria (excluding MODS). Sensitivity by patient against clinical gold standard for ZN smear, MODS MGIT and LJ were 52.6%, 64.9%, 70.2% and 70.2%, respectively. Specificity of all microbiological techniques was 100%. Positive and negative predictive values for MODS were 100% and 78.7%, respectively for HIV infected patients and 100% and 82.1% for HIV negative patients. The median time to positive was 6 days (interquartile range 5–7), significantly faster than MGIT at 15.5 days (interquartile range 12–24), and LJ at 24 days (interquartile range 18–35 days) (P<0.01).

Conclusions

We have shown MODS to be a sensitive, rapid technique for the diagnosis of TBM with high sensitivity, ease of performance and low cost (0.53 USD/sample).     

Discovering robust protein biomarkers for disease from relative expression reversals in 2-D DIGE data

This study assesses the ability of a novel family of machine learning algorithms to identify changes in relative protein expression levels, measured using 2-D DIGE data, which support accurate class prediction. The analysis was done using a training set of 36 total cellular lysates comprised of six normal and three cancer biological replicates (the remaining are technical replicates) and a validation set of four normal and two cancer samples. Protein samples were separated by 2-D DIGE and expression was quantified using DeCyder-2D Differential Analysis Software. The relative expression reversal (RER) classifier correctly classified 9/9 training biological samples (p<0.022) as estimated using a modified version of leave one out cross validation and 6/6 validation samples. The classification rule involved comparison of expression levels for a single pair of protein spots, tropomyosin isoforms and alpha-enolase, both of which have prior association as potential biomarkers in cancer. The data was also analyzed using algorithms similar to those found in the extended data analysis package of DeCyder software. We propose that by accounting for sources of within- and between-gel variation, RER classifiers applied to 2-D DIGE data provide a useful approach for identifying biomarkers that discriminate among protein samples of interest.     

Minimum population size,genetic diversity,and social structure of the Asian elephant in Cat Tien National Park and its adjoining areas,Vietnam, based on molecular genetic analyses

Vietnam’s elephant population that has suffered severe declines during the past three decades is now believed to number 60–80 individuals in the wild. Cat Tien National Park is thought to be one of the key areas for the recovery of Vietnam’s elephants. We carried out a molecular genetic study of elephants in Cat Tien National Park and its adjoining areas with the objectives of estimating minimum population size, assessing genetic diversity, and obtaining insights into social organization. We obtained a minimum population size of 11 elephants based on a combination of unique nuclear microsatellite genotypes and mitochondrial haplotypes. While mitochondrial diversity based on a 600-base pair segment was high in this small sample of individuals, the six microsatellite loci examined showed low diversity and the signature of a recent population bottleneck. Along with nuclear genetic depauperation of Cat Tien’s elephants, we also report disruption of normal social organization, with different matrilines having coalesced into a single social group because of anthropogenic disturbance. The results emphasize the critical condition of this elephant population and the need for urgent conservation measures if this population is to be saved.     

Potent effect of target structure on microRNA function

MicroRNAs (miRNAs) are small noncoding RNAs that repress protein synthesis by binding to target messenger RNAs. We investigated the effect of target secondary structure on the efficacy of repression by miRNAs. Using structures predicted by the Sfold program, we model the interaction between an miRNA and a target as a two-step hybridization reaction: nucleation at an accessible target site followed by hybrid elongation to disrupt local target secondary structure and form the complete miRNA-target duplex. This model accurately accounts for the sensitivity to repression by let-7 of various mutant forms of the Caenorhabditis elegans lin-41 3' untranslated region and for other experimentally tested miRNA-target interactions in C. elegans and Drosophila melanogaster. These findings indicate a potent effect of target structure on target recognition by miRNAs and establish a structure-based framework for genome-wide identification of animal miRNA targets.     

中印度洋与南海西部表层海水细菌多样性

细菌在海洋生物地球化学循环中发挥着重要作用。为更好地了解海洋细菌的特征及其在海洋环境中的潜在作用, 本文利用纯培养与16S rRNA基因高通量测序技术对中印度洋与南海西部海域表层海水细菌多样性进行研究。纯培养结果表明, 自中印度洋与南海西部表层海水中共分离275株可培养海洋细菌, 隶属于4门49属75种。变形菌门是绝对优势类群(占总株数的68.7%), 其次是放线菌门(21.5%)、拟杆菌门(9.1%)和厚壁菌门(0.7%)。在属水平, 微杆菌属(Microbacterium)与弧菌属(Vibrio)是主要的优势属, 共占总株数的30.0%。在3种分离培养基中, 自1/10 × 2216E培养基中分离细菌的数目与种类最多(89株, 30属); 分离菌株中的细菌菌株有7、9与3个属分别仅在2216E、1/10 × 2216E及葡萄糖甘露糖(glucose-mannose, GM)培养基中生长。此外, 共分离培养出50株细菌(26种)可能代表潜在新分类单元。高通量测序结果显示, 中印度洋和南海西部表层海水中共有23个门531个属。优势门类为变形菌门(72.2%)和拟杆菌门(15.3%), 优势属为嗜冷杆菌属(Psychrobacter, 24.4%)、盐单胞菌属(Halomonas, 16.3%)和亚硫酸杆菌属(Sulfitobacter, 13.9%)。此外, 中印度洋表层海水细菌Shannon-Wiener指数与Pielou均匀度指数显著高于南海西部(P < 0.05), 且细菌群落结构显著不同(P < 0.05)。综合纯培养与原位细菌数据得出, 中印度洋与南海西部海洋细菌具有丰富的多样性, 具有进一步开发研究的价值。     

When did the ancestor of true bugs become stinky? Disentangling the phylogenomics of Hemiptera–Heteroptera

The phylogeny of true bugs (Hemiptera: Heteroptera), one of the most diverse insect groups in terms of morphology and ecology, has been the focus of attention for decades with respect to several deep nodes between the suborders of Hemiptera and the infraorders of Heteroptera. Here, we assembled a phylogenomic data set of 53 taxa and 3102 orthologous genes to investigate the phylogeny of Hemiptera–Heteroptera, and both concatenation and coalescent methods were used. A binode-control approach for data filtering was introduced to reduce the incongruence between different genes, which can improve the performance of phylogenetic reconstruction. Both hypotheses (Coleorrhyncha + Heteroptera) and (Coleorrhyncha + Auchenorrhyncha) received support from various analyses, in which the former is more consistent with the morphological evidence. Based on a divergence time estimation performed on genes with a strong phylogenetic signal, the origin of true bugs was dated to 290–268 Ma in the Permian, the time in Earth's history with the highest concentration of atmospheric oxygen. During this time interval, at least 1007 apomorphic amino acids were retained in the common ancestor of the extant true bugs. These molecular apomorphies are located in 553 orthologous genes, which suggests the common ancestor of the extant true bugs may have experienced large-scale evolution at the genome level.     

Phosphoinositide-dependent kinase 1–associated glycolysis is regulated by miR-409-3p in clear cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) is the most popular kidney cancer in adults. Metabolic shift toward aerobic glycolysis is a fundamental factor for ccRCC therapy. MicroRNAs (miRNAs) are thought to be important regulators in ccRCC development and progression. Phosphoinositide-dependent kinase 1 (PDK1) is required for metabolic activation; however, the role of PDK1-induced glycolytic metabolism regulated by miRNAs is unclear in ccRCC. So, the purpose of the current study is to elucidate the underlying mechanism in ccRCC cell metabolism mediated by PDK1. Our results revealed that miR-409-3p inhibited glycolysis by regulating PDK1 expression in ccRCC cells. We also found that miR-409-3p was regulated by hypoxia. Our results indicated that PDK1 facilitated ccRCC cell glycolysis, regulated by miR-409-3p in hypoxia.     

果肉和埋土深度对新疆野杏种子萌发与幼苗生长的影响

为探明新疆野杏（Armeniaca vulgaris）种子萌发与幼苗生长对果肉和埋土深度的响应,以期为新疆野杏的天然更新与实生苗培育提供理论参考。通过2种果皮结构（有果肉和无果肉）的种子在不同埋土深度（地表至18.0 cm的14个梯度）对新疆野杏种子萌发和幼苗生长进行研究,旨在揭示果皮结构和埋土深度对新疆野杏种子萌发与成苗能力的影响。结果表明：果肉和埋土深度显著影响野杏种子的萌发、幼苗生长与质量（P<0.05）。埋土深度<3.0 cm不利于成苗,埋土深度>6.0 cm时,萌发能力与幼苗生长量随埋土深度的增加而降低,3.0～6.0cm为适宜埋土深度。无果肉种子萌发优于有果肉种子,萌发率、萌发指数、成苗率、活力指数分别增长了37.18%、3.88%、37.18%、26.59%,幼苗高、基径、叶片数量、根冠比、幼苗质量指数分别增长了36.99%、7.48%、68.69%、20.61%、14.29%,其萌发能力与幼苗生长量显著高于有果肉种子（P<0.05）。有无果肉种子的萌发和幼苗生长指标与埋土深度呈显著负相关（P<0.05）。研究结果表明,无果肉处理对新疆野杏种...