Long-term results of endoscopic-assisted cranionasal resection for olfactory neuroblastoma - single centre experience of fourteen patients

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Developing a platform of environmental omics for the green-lipped mussel Perna viridis

Session Track: Aquatic and terrestrial ecotoxicologyOral presentationConference Theme: Science across bridges, borders and boundariesThe green-lipped mussel Perna viridis is an important marine biomonitor species in pollution monitoring and ecotoxicological studies in Asia-Pacific region, and considered as a subtropical equivalent biomonitor of the temperate Mytilus species. However, the genomic information of P. viridis is still largely unexplored when compared with Mytilus species. This study aimed to establish a transcriptomic profile of P. viridis using the next generation sequencing technology and provide a good representative set of genomic information for elucidation of toxic mechanisms upon pollution stresses and identification of a suite of suitable biomarkers for monitoring marine pollution and environmental stresses. To obtain a wide spectrum of environmental-associated transcripts, adult mussels (4-5 cm shell length) were collected from different locations in Hong Kong and from those after 24-hour exposures to various challenges of physical stresses and chemical pollutants, so as to cover a wide range of stress-associated transcription patterns for future environmental studies. Two males and females from each location and from each treatment were chosen for obtaining the three target tissues (i.e., hepatopancreas, gill and adductor muscle). For each sex and each tissue type, a total RNA sample was extracted from pooled tissues from the field and laboratory treated mussels. The RNA sample was subjected to cDNA library construction, followed by the RNA-sequencing using a Solexa GAIIx (Illumina). Including the splicing variants, a de novo assembly of a total length of 295,064,579 base-pair (bp) contig was obtained, with 233,257 contigs assembled of an average size of 1264 bp. The 192,879 non-redundant assembled transcripts were blasted against the NCBI nr database and three molluscan EST databases, and resulted in 44,713 transcripts with at least a blast hit, and having a top match with the sequences from the Pacific oyster, Crassostrea gigas (27,651 transcripts). A total of 5,131 transcripts were assigned with KEGG annotation involving in 329 pathways. Based on multivariate statistical analysis, expression patterns of genes from stress associated responses and detoxification were strongly tissue-specific but the differences between genders were little. The anticipated genomic database generated from this study will further strengthen the role of P. viridis as a universal marine biomonitor in the Asia-Pacific region.published_or_final_versio

Comparison of Mutation Patterns in Full-Genome A/H3N2 Influenza Sequences Obtained Directly from Clinical Samples and the Same Samples after a Single MDCK Passage

Human influenza viruses can be isolated efficiently from clinical samples using Madin-Darby canine kidney (MDCK) cells. However, this process is known to induce mutations in the virus as it adapts to this non-human cell-line. We performed a systematic study to record the pattern of MDCK-induced mutations observed across the whole influenza A/H3N2 genome. Seventy-seven clinical samples collected from 2009-2011 were included in the study. Two full influenza genomes were obtained for each sample: one from virus obtained directly from the clinical sample and one from the matching isolate cultured in MDCK cells. Comparison of the full-genome sequences obtained from each of these sources showed that 42% of the 77 isolates had acquired at least one MDCK-induced mutation. The presence or absence of these mutations was independent of viral load or sample origin (in-patients versus out-patients). Notably, all the five hemagglutinin missense mutations were observed at the hemaggutinin 1 domain only, particularly within or proximal to the receptor binding sites and antigenic site of the virus. Furthermore, 23% of the 77 isolates had undergone a MDCK-induced missense mutation, D151G/N, in the neuraminidase segment. This mutation has been found to be associated with reduced drug sensitivity towards the neuraminidase inhibitors and increased viral receptor binding efficiency to host cells. In contrast, none of the neuraminidase sequences obtained directly from the clinical samples contained the D151G/N mutation, suggesting that this mutation may be an indicator of MDCK culture-induced changes. These D151 mutations can confound the interpretation of the hemagglutination inhibition assay and neuraminidase inhibitor resistance results when these are based on MDCK isolates. Such isolates are currently in routine use in the WHO influenza vaccine and drug-resistance surveillance programs. Potential data interpretation miscalls can therefore be avoided by careful exclusion of such D151 mutants after further sequence analysis.published_or_final_versio

Albuminuria is a marker of increasing intracranial and extracranial vascular involvement in Type 2 diabetic Chinese patients

AIMS/HYPOTHESIS: Albuminuria has been reported to be a marker of cardiovascular risk factors and disease morbidity and mortality, but its relationship with intracerebral atherosclerotic disease is less clear. The aim of this study was to investigate the association between albuminuria and intracranial and extracranial vascular involvement in Chinese Type 2 diabetic patients. METHODS: The anthropometric and fasting biochemical measurements of 966 Type 2 diabetic patients with normoalbuminuria (55.6%), microalbuminuria (27.7%) or macroalbuminuria (16.7%) were compared. The prevalence of microvascular and macrovascular disease and middle cerebral artery (MCA) stenosis, measured by transcranial Doppler ultrasound, were also compared between the groups. RESULTS: Albuminuria was closely associated with a range of adverse parameters, including high BP, dyslipidaemia, smoking and adiposity (all p<0.01). The prevalence of microvascular disease (retinopathy p<0.001) and macrovascular disease (peripheral vascular disease p=0.012, myocardial infarction, p=0.004, MCA stenosis p<0.001) increased significantly with increasing levels of albuminuria. Albuminuria was also found to be an independent predictor of microvascular and macrovascular disease. CONCLUSIONS/INTERPRETATION: Albuminuria was an independent predictor of increasing levels of vascular risk factors and microvascular and macrovascular disease in this group of Type 2 diabetic patients, and a possible role for albuminuria as a marker of intracranial cerebrovascular disease should be further investigated.postprin

De novo transcriptome analysis of Perna viridis highlights tissue-specific patterns for environmental studies

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Pulmonary artery sarcoma diagnosed by endobronchial ultrasound-guided transbronchial needle aspiration

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Fungal Communities in Sediments Along a Depth Gradient in the Eastern Tropical Pacific

Deep waters represent the largest biome on Earth and the largest ecosystem of Costa Rica. Fungi play a fundamental role in global biogeochemical cycling in marine sediments, yet, they remain little explored. We studied fungal diversity and community composition in several marine sediments from 16 locations sampled along a bathymetric gradient (from a depth of 380 to 3,474 m) in two transects of about 1,500 km length in the Eastern Tropical Pacific (ETP) of Costa Rica. Sequence analysis of the V7-V8 region of the 18S rRNA gene obtained from sediment cores revealed the presence of 787 fungal amplicon sequence variants (ASVs). On average, we detected a richness of 75 fungal ASVs per sample. Ascomycota represented the most abundant phylum with Saccharomycetes constituting the dominant class. Three ASVs accounted for ca. 63% of all fungal sequences: the yeast Metschnikowia (49.4%), Rhizophydium (6.9%), and Cladosporium (6.7%). We distinguished a cluster composed mainly by yeasts, and a second cluster by filamentous fungi, but we were unable to detect a strong effect of depth and the overlying water temperature, salinity, dissolved oxygen (DO), and pH on the composition of fungal communities. We highlight the need to understand further the ecological role of fungi in deep-sea ecosystems.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Ciencias del Mar y Limnología (CIMAR)UCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Biologí

Germline CRISPR/Cas9-mediated gene editing prevents vision loss in a novel mousemodel of Aniridia.

Aniridia is a rare eye disorder, which is caused by mutations in the paired box 6 (PAX6) gene and results in vision loss due to the lack of a long-term vision-saving therapy. One potential approach to treating aniridia is targeted CRISPR-based genome editing. To enable the Pax6 small eye (Sey) mouse model of aniridia, which carries the same mutation found in patients, for preclinical testing of CRISPR-based therapeutic approaches, we endogenously tagged the Sey allele, allowing for the differential detection of protein from each allele. We optimized a correction strategy in vitro then tested it in vivo in the germline of our new mouse to validate the causality of the Sey mutation. The genomic manipulations were analyzed by PCR, as well as by Sanger and next-generation sequencing. The mice were studied by slit lamp imaging, immunohistochemistry, and western blot analyses. We successfully achieved both in vitro and in vivo germline correction of the Sey mutation, with the former resulting in an average 34.8% ± 4.6% SD correction, and the latter in restoration of 3xFLAG-tagged PAX6 expression and normal eyes. Hence, in this study we have created a novel mouse model for aniridia, demonstrated that germline correction of the Sey mutation alone rescues the mutant phenotype, and developed an allele-distinguishing CRISPR-based strategy for aniridia

Overexpression of protein kinase C-beta 1 isoenzyme suppresses SC-236-induced apoptosis in gastric epithelial cells

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