Construction and characterization of an infectious clone of coxsackievirus A16

<p>Abstract</p> <p>Background</p> <p>Coxsackievirus A16 (CVA16) is a member of the <it>Enterovirus </it>genus of the <it>Picornaviridae </it>family and it is a major etiological agent of hand, foot, and mouth disease (HFMD), which is a common illness affecting children. CVA16 possesses a single-stranded positive-sense RNA genome containing approximately 7410 bases. Current understanding of the replication, structure and virulence determinants of CVA16 is very limited, partly due to difficulties in directly manipulating its RNA genome.</p> <p>Results</p> <p>Two overlapping cDNA fragments were amplified by RT-PCR from the genome of the shzh05-1 strain of CVA16, encompassing the nucleotide regions 1-4392 and 4381-7410, respectively. These two fragments were then joined <it>via </it>a native <it>Xba</it>I site to yield a full-length cDNA. A T7 promoter and poly(A) tail were added to the 5' and 3' ends, respectively, forming a full CVA16 cDNA clone. Transfection of RD cells <it>in vitro </it>with RNA transcribed directly from the cDNA clone allowed the recovery of infectious virus in culture. The CVA16 virus recovered from these cultures was functionally and genetically identical to its parent strain.</p> <p>Conclusions</p> <p>We report the first construction and characterization of an infectious cDNA clone of CVA16. The availability of this infectious clone will greatly enhance future virological investigations and vaccine development for CVA16.</p

Patterns of MicroRNA Expression in Normal and Early Alzheimer\u27s Disease Human Temporal Cortex: White Matter versus Gray Matter

MicroRNA (miRNA) expression was assessed in human cerebral cortical gray matter (GM) and white matter (WM) in order to provide the first insights into the difference between GM and WM miRNA repertoires across a range of Alzheimer\u27s disease (AD) pathology. RNA was isolated separately from GM and WM portions of superior and middle temporal cerebral cortex (N = 10 elderly females, postmortem interval \u3c 4 h). miRNA profiling experiments were performed using state-of-the-art Exiqon© LNA-microarrays. A subset of miRNAs that appeared to be strongly expressed according to the microarrays did not appear to be conventional miRNAs according to Northern blot analyses. Some well-characterized miRNAs were substantially enriched in WM as expected. However, most of the miRNA expression variability that correlated with the presence of early AD-related pathology was seen in GM. We confirm that downregulation of a set of miRNAs in GM (including several miR-15/107 genes and miR-29 paralogs) correlated strongly with the density of diffuse amyloid plaques detected in adjacent tissue. A few miRNAs were differentially expressed in WM, including miR-212 that is downregulated in AD and miR-424 which is upregulated in AD. The expression of certain miRNAs correlates with other miRNAs across different cases, and particular subsets of miRNAs are coordinately expressed in relation to AD-related pathology. These data support the hypothesis that patterns of miRNA expression in cortical GM may contribute to AD pathogenetically, because the aggregate change in miRNA expression observed early in the disease would be predicted to cause profound changes in gene expression

Efficient eradication of organotin utilizing K2FeO4 augmented by nZVI: revelations on influential factors, kinetic dynamics, and mechanistic insights

In this study, we investigated the oxidative mechanisms of nano zero-valent iron (nZVI) enhanced potassium ferrate (K2FeO4), focusing on tributyltin (TBT) and triphenyltin (TPhT) as target pollutants. The addition of nZVI enhanced the degradation of both organic tin compounds by K2FeO4, exhibiting pseudo-first-order kinetics influenced by pH, carbonate, and fulvic acid. nZVI, as a reducing agent, facilitated the generation of stronger oxidizing species Fe (V) and Fe (IV) from K2FeO4 through electron transfer. The presence of hydroxyl radicals (OH) was confirmed by tert-butyl alcohol (TBA) verification. Intermediate products of TBT degradation by nZVI-enhanced K2FeO4 were identified using GC-MS, confirming de-alkylation leading to stepwise oxidation to inorganic tin ions. Due to excessively long Sn-C bonds in diphenyltin, rendering them unstable, Density Functional Theory (DFT) calculations were employed. Results indicated that Fe (IV) and Fe (V) predominantly attacked the Sn-C bonds of TPhT, while OH primarily targeted the benzene ring. HOMO energy levels suggested that Fe (V) was more prone to oxidizing TPhT than Fe (IV). Gibbs free energy calculations demonstrated that, in the presence of Fe (IV) and Fe (V), the energy barrier for breaking bonds and oxidizing into inorganic tin ions decreased, favoring the process over the self-decomposition of TPhT. Additionally, the lower energy barrier for OH indicated an easier degradation of TPhT. This study reveals that nZVI-enhanced K2FeO4 effectively removes TBT and TPhT, contributing to the understanding of the ferrate-mediated degradation mechanism of organic tin compounds. The findings offer insights and theoretical guidance for remediating organic tin pollution in aquatic environments

The Expression of MicroRNA miR-107 Decreases Early in Alzheimer\u27s Disease and May Accelerate Disease Progression through Regulation of β-Site Amyloid Precursor Protein-Cleaving Enzyme 1

MicroRNAs (miRNAs) are small regulatory RNAs that participate in posttranscriptional gene regulation in a sequence-specific manner. However, little is understood about the role(s) of miRNAs in Alzheimer\u27s disease (AD). We used miRNA expression microarrays on RNA extracted from human brain tissue from the University of Kentucky Alzheimer\u27s Disease Center Brain Bank with near-optimal clinicopathological correlation. Cases were separated into four groups: elderly nondemented with negligible AD-type pathology, nondemented with incipient AD pathology, mild cognitive impairment (MCI) with moderate AD pathology, and AD. Among the AD-related miRNA expression changes, miR-107 was exceptional because miR-107 levels decreased significantly even in patients with the earliest stages of pathology. In situ hybridization with cross-comparison to neuropathology demonstrated that particular cerebral cortical laminas involved by AD pathology exhibit diminished neuronal miR-107 expression. Computational analysis predicted that the 3′-untranslated region (UTR) of β-site amyloid precursor protein-cleaving enzyme 1 (BACE1) mRNA is targeted multiply by miR-107. From the same RNA material analyzed on miRNA microarrays, mRNA expression profiling was performed using Affymetrix Exon Array microarrays on nondemented, MCI, and AD patients. BACE1 mRNA levels tended to increase as miR-107 levels decreased in the progression of AD. Cell culture reporter assays performed with a subset of the predicted miR-107 binding sites indicate the presence of at least one physiological miR-107 miRNA recognition sequence in the 3′-UTR of BACE1 mRNA. Together, the coordinated application of miRNA profiling, Affymetrix microarrays, new bioinformatics predictions, in situ hybridization, and biochemical validation indicate that miR-107 may be involved in accelerated disease progression through regulation of BACE1

Vegetation greenness and photosynthetic phenology in response to climatic determinants

Vegetation phenology is a key indicator of vegetation-climate interactions and carbon sink changes in ecosystems. Therefore, it is very important to understand the temporal and spatial variability of vegetation phenology and the driving climatic determinants [e.g., temperature (Ts) and soil moisture (SM)]. Vegetation greenness and photosynthetic phenology were derived using the double logistic (DL) method to enhance vegetation index (EVI) and solar-induced chlorophyll fluorescence (SIF) spring and autumn phenology, respectively. The growing season length (GSL) of greenness phenology (about 100 days) derived EVI was longer than GSL of photosynthetic phenology (about 80 days) derived SIF. Although their overall spatiotemporal pattern trends were consistent, photosynthetic phenology varied 1.4 to 3.1 times more than greenness phenology over time. In addition, SIF-based photosynthetic phenology and EVI-based greenness phenology showed consistent factors of drivers but differed to some extent in spatial patterns and the most relevant preseason dates. Spring photosynthetic phenology was mainly influenced by pre-season mean cumulative Ts (about 90 days). However, greenness phenology was controlled by both pre-seasons mean cumulative Ts [(about 55 days) and mean cumulative SM (about 40 days)]. Autumn photosynthetic phenology was controlled by both periods’ mean cumulative Ts [(about 20 days) and SM (about 20 days)], but autumn greenness phenology was mainly influenced by pre-season mean cumulative Ts (85 days). The comparison analysis of SIF and EVI phenology helps to understand the difference between photosynthetic phenology and greenness phenology at a regional scale

miRNA-378 reverses chemoresistance to cisplatin in lung adenocarcinoma cells by targeting secreted clusterin

Cisplatin resistance is a major obstacle in the treatment of NSCLC, and its mechanism has not been fully elucidated. The objectives of the study were to determine the role of miR-378 in the sensitivity of lung adenocarcinoma cells to cisplatin (cDDP) and its working mechanism. With TargetScan and luciferase assay, miR-378 was found to directly target sCLU. miR-378 and sCLU were regulated in A549/cDDP and Anip973/cDDP cells to investigate the effect of miR-378 on the sensitivity and apoptotic effects of cDDP. The effect of miR-378 upregulation on tumor growth was analyzed in a nude mouse xenograft model. The correlation between miR-378 and chemoresistance was tested in patient samples. We found that upregulation of miR-378 in A549/cDDP and Anip973/cDDP cells significantly down-regulated sCLU expression, and sensitized these cells to cDDP. miR-378 overexpression inhibited tumor growth and sCLU expression in a xenograft animal model. Analysis of human lung adenocarcinoma tissues revealed that the cDDP sensitive group expressed higher levels of miR-378 and lower levels of sCLU. miR-378 and sCLU were negatively correlated. To conclude, we identified sCLU as a novel miR-378 target, and we showed that targeting sCLU via miR-378 may help disable the chemoresistance against cisplatin in lung adenocarcinoma cells

Yeast-produced subunit protein vaccine elicits broadly neutralizing antibodies that protect mice against Zika virus lethal infection

International audienceZika virus (ZIKV) infection is a serious public health concern due to its ability to induce neurological defects and its potential for rapid transmission at a global scale. However, no vaccine is currently available to prevent ZIKV infection. Here, we report the development of a yeast-derived subunit protein vaccine for ZIKV. The envelope protein domain III (EDIII) of ZIKV was produced as a secretory protein in the yeast Pichia pastoris. The yeast-derived EDIII could inhibit ZIKV infection in vitro in a dose-dependent manner, suggesting that it had acquired an appropriate conformation to bind to cellular receptors of ZIKV. Immunization with recombinant EDIII protein effectively induced antigen-specific binding antibodies and cellular immune responses. The resulting anti-EDIII sera could efficiently neutralize ZIKV representative strains from both Asian and African lineages. Passive transfer with the anti-EDIII neutralizing sera could confer protection against lethal ZIKV challenge in mice. Importantly, we found that purified anti-EDIII antibodies did not cross-react with closely related dengue virus (DENV) and therefore did not enhance DENV infection. Collectively, our results demonstrate that yeast-produced EDIII is a safe and effective ZIKV vaccine candidate

Novel Human \u3cem\u3eABCC9/SUR2\u3c/em\u3e Brain-Expressed Transcripts and an eQTL Relevant to Hippocampal Sclerosis of Aging

ABCC9 genetic polymorphisms are associated with increased risk for various human diseases including hippocampal sclerosis of aging. The main goals of this study were 1 \u3e to detect the ABCC9 variants and define the specific 3′ untranslated region (3′UTR) for each variant in human brain, and 2 \u3e to determine whether a polymorphism (rs704180) associated with risk for hippocampal sclerosis of aging pathology is also associated with variation in ABCC9 transcript expression and/or splicing. Rapid amplification of ABCC9 cDNA ends (3′RACE) provided evidence of novel 3′ UTR portions of ABCC9 in human brain. In silico and experimental studies were performed focusing on the single nucleotide polymorphism, rs704180. Analyses from multiple databases, focusing on rs704180 only, indicated that this risk allele is a local expression quantitative trait locus (eQTL). Analyses of RNA from human brains showed increased ABCC9 transcript levels in individuals with the risk genotype, corresponding with enrichment for a shorter 3′ UTR which may be more stable than variants with the longer 3′ UTR. MicroRNA transfection experiments yielded results compatible with the hypothesis that miR‐30c causes down‐regulation of SUR2 transcripts with the longer 3′ UTR. Thus we report evidence of complex ABCC9 genetic regulation in brain, which may be of direct relevance to human disease