Assessing diversity of the female urine microbiota by high throughput sequencing of 16S rDNA amplicons

<p>Abstract</p> <p>Background</p> <p>Urine within the urinary tract is commonly regarded as "sterile" in cultivation terms. Here, we present a comprehensive in-depth study of bacterial 16S rDNA sequences associated with urine from healthy females by means of culture-independent high-throughput sequencing techniques.</p> <p>Results</p> <p>Sequencing of the V1V2 and V6 regions of the 16S ribosomal RNA gene using the 454 GS FLX system was performed to characterize the possible bacterial composition in 8 culture-negative (<100,000 CFU/ml) healthy female urine specimens. Sequences were compared to 16S rRNA databases and showed significant diversity, with the predominant genera detected being <it>Lactobacillus</it>, <it>Prevotella </it>and <it>Gardnerella</it>. The bacterial profiles in the female urine samples studied were complex; considerable variation between individuals was observed and a common microbial signature was not evident. Notably, a significant amount of sequences belonging to bacteria with a known pathogenic potential was observed. The number of operational taxonomic units (OTUs) for individual samples varied substantially and was in the range of 20 - 500.</p> <p>Conclusions</p> <p>Normal female urine displays a noticeable and variable bacterial 16S rDNA sequence richness, which includes fastidious and anaerobic bacteria previously shown to be associated with female urogenital pathology.</p

Overview of the Nordic Seas CARINA data and salinity measurements

Water column data of carbon and carbon relevant hydrographic and hydrochemical parameters from 188 previously non-publicly available cruises in the Arctic, Atlantic, and Southern Ocean have been retrieved and merged into a new database: CARINA (CARbon IN the Atlantic). The data have been subject to rigorous quality control (QC) in order to ensure highest possible quality and consistency. The data for most of the parameters included were examined in order to quantify systematic biases in the reported values, i.e. secondary quality control. Significant biases have been corrected for in the data products, i.e. the three merged files with measured, calculated and interpolated values for each of the three CARINA regions; the Arctic Mediterranean Seas (AMS), the Atlantic (ATL) and the Southern Ocean (SO). With the adjustments the CARINA database is consistent both internally as well as with GLODAP (Key et al., 2004) and is suitable for accurate assessments of, for example, oceanic carbon inventories and uptake rates and for model validation. The Arctic Mediterranean Seas include the Arctic Ocean and the Nordic Seas, and the quality control was carried out separately in these two areas. This contribution provides an overview of the CARINA data from the Nordic Seas and summarises the findings of the QC of the salinity data. One cruise had salinity data that were of questionable quality, and these have been removed from the data product. An evaluation of the consistency of the quality controlled salinity data suggests that they are consistent to at least ±0.005

Inorganic carbon and water masses in the Irminger Sea since 1991

The subpolar region in the North Atlantic is a major sink for anthropogenic carbon. While the storage rates show large interannual variability related to atmospheric forcing, less is known about variability in the natural dissolved inorganic carbon (DIC) and the combined impact of variations in the two components on the total DIC inventories. Here, data from 15 cruises in the Irminger Sea covering the 24-year period between 1991 and 2015 were used to determine changes in total DIC and its natural and anthropogenic components. Based on the results of an extended optimum multiparameter analysis (eOMP), the inventory changes are discussed in relation to the distribution and evolution of the main water masses. The inventory of DIC increased by 1.43±0.17 molm-2 yr-1 over the period, mainly driven by the increase in anthropogenic carbon (1.84±0.16 molm-2 yr-1) but partially offset by a loss of natural DIC (-0.5±0.22 molm-2 yr-1). Changes in the carbon storage rate can be driven by concentration changes in the water column, for example due to the ageing of water masses, or by changes in the distribution of water masses with different concentrations either by local formation or advection. A decomposition of the trends into their main drivers showed that variations in natural DIC inventories are mainly driven by changes in the layer thickness of the main water masses, while anthropogenic carbon is most affected by concentration changes. The storage rates of anthropogenic carbon are sensitive to data selection, while changes in DIC inventory show a robust signal on short timescales associated with the strength of convection

Respiration Patterns in the Dark Ocean

In the dark ocean, respiring organisms are the main sink for dissolved oxygen. The respiration rate in a given seawater volume can be quantified through dissolved oxygen drawdown or organic matter consumption as a function of time. Estimates of dissolved oxygen utilization rates (OUR) abound in the literature, but are typically obtained using proxies of questionable accuracy, often with low vertical resolution, and neglecting key regions such as the Southern and Indian oceans. Respiration rates based on particulate (POC) or dissolved (DOC) organic carbon are also sparsely observed and for DOC are unavailable in many regions. Consequently, the relative contributions of POC or DOC as a respiration substrate in the dark ocean are unknown. Here, we use recent datasets of true oxygen utilization, seawater age, and DOC to derive OUR and DOC consumption-rate profiles in 10 oceanic regions. We demonstrate that although DOC and POC consumption rates are globally consistent with OUR, they underestimate OUR in the deep, suggesting strong oxygen utilization at the seafloor. In the abyss, we find a negative correlation of the DOC consumption rate with seawater age, suggesting that DOC reactivity decreases along the deep branch of the conveyor circulation. Our results highlight that benthic organisms are sensitive to perturbations in the surface production of organic matter and to large-scale circulation changes that affect its supply to the abyss

Endothelial Surface Layer Degradation by Chronic Hyaluronidase Infusion Induces Proteinuria in Apolipoprotein E-Deficient Mice

Functional studies show that disruption of endothelial surface layer (ESL) is accompanied by enhanced sensitivity of the vasculature towards atherogenic stimuli. However, relevance of ESL disruption as causal mechanism for vascular dysfunction remains to be demonstrated. We examined if loss of ESL through enzymatic degradation would affect vascular barrier properties in an atherogenic model. Eight week old male apolipoprotein E deficient mice on Western-type diet for 10 weeks received continuous active or heat-inactivated hyaluronidase (10 U/hr, i.v.) through an osmotic minipump during 4 weeks. Blood chemistry and anatomic changes in both macrovasculature and kidneys were examined. Infusion with active hyaluronidase resulted in decreased ESL (0.32±0.22 mL) and plasma volume (1.03±0.18 mL) compared to inactivated hyaluronidase (0.52±0.29 mL and 1.28±0.08 mL, p<0.05 respectively).Active hyaluronidase increased proteinuria compared to inactive hyaluronidase (0.27±0.02 vs. 0.15±0.01 µg/µg protein/creatinin, p<0.05) without changes in glomerular morphology or development of tubulo-interstitial inflammation. Atherosclerotic lesions in the aortic branches showed increased matrix production (collagen, 32±5 vs. 18±3%; glycosaminoglycans, 11±5 vs. 0.1±0.01%, active vs. inactive hyaluronidase, p<0.05). ESL degradation in apoE deficient mice contributes to reduced increased urinary protein excretion without significant changes in renal morphology. Second, the induction of compositional changes in atherogenic plaques by hyaluronidase point towards increased plaque vulnerability. These findings support further efforts to evaluate whether ESL restoration is a valuable target to prevent (micro) vascular disease progressio

Role of Glomerular Proteoglycans in IgA Nephropathy

Mesangial matrix expansion is a prominent feature of the most common form of glomerulonephritis, IgA nephropathy (IgAN). To find molecular markers and improve the understanding of the disease, the gene and protein expression of proteoglycans were investigated in biopsies from IgAN patients and correlated to clinical and morphological data. We collected and microdissected renal biopsies from IgAN patients (n = 19) and from healthy kidney donors (n = 14). Patients were followed for an average time of 4 years and blood pressure was according to target guidelines. Distinct patterns of gene expression were seen in glomerular and tubulo-interstitial cells. Three of the proteoglycans investigated were found to be of special interest and upregulated in glomeruli: perlecan, decorin and biglycan. Perlecan gene expression negatively correlated to albumin excretion and progress of the disease. Abundant decorin protein expression was found in sclerotic glomeruli, but not in unaffected glomeruli from IgAN patients or in controls. Transforming growth factor beta (TGF-β), known to interact with perlecan, decorin and biglycan, were upregulated both on gene and protein level in the glomeruli. This study provides further insight into the molecular mechanisms involved in mesangial matrix expansion in IgAN. We conclude that perlecan is a possible prognostic marker for patients with IgAN. In addition, the up-regulation of biglycan and decorin, as well as TGF-β itself, indicate that regulation of TGF-β, and other profibrotic markers plays a role in IgAN pathology

Oceanic biogeochemical characteristic maps identified with holistic use of satellite, model and data

This is the final published version.Ocean province level plankton community exhibit heterogeneity across Arctic, Nordic, Atlantic Gyre and Southern Ocean provinces. GreenSeas research is an international FP7 consortium that includes Arctic, Atlantic and Southern Ocean based research teams who are analysing the planktonic ecosystem. We are looking at how the planktonic ecosystem responds to environmental and climate change. Using Earth Observation monitoring data we report new results on identifying generic plankton characteristics observable at a province level, and also touch on spatial and temporal trends that are evident using a holistic analysis framework. Using advanced statistical methods this framework compares and combines Earth Observation information together with an in-situ Oceanic plankton Analytical Database and up to 40 year ocean general circulation biogeochemical model (OGCBM) time series of the equivalent plankton and sea-state measures of this system. Specifically, we outline the use of the GreenSeas Analytical Database, which is a harmonised set of Oceanic in-situ plankton and sea-state measures covering different cruises and time periods. The Analytical Database information ranges from plankton community, primary production, nutrient cycling to physical sea state temperature and salinity measures. The combined analysis utilises current, 10 year+ Earth Observations of ocean colour and sea surface temperature metrics and interprets these together with biogeochemical model outputs from PELAGOS, ERSEM & NORWECOM model runs to help identify planktonic based biomes. Generic planktonic characteristic maps that are equivalently observable in both the Earth Observations and numerical models are reported on. Both ocean surface and sub-surface signals are analysed together with relevant Analytical Database biome extracts. We present the current results of this inter-comparison & discuss challenges of identifying the province level plankton dominance with the satellite, model and data. In particular we discuss the strategic importance of systematically analysing the knowledge present in the existing key long term Oceanic observation platforms through such holistic analysis frameworks. These maps help to enhance and improve current biogeochemical models, our understanding of the plankton community structure and predictions used for future assessment of climate change

A human glomerular SAGE transcriptome database

Background: To facilitate in the identification of gene products important in regulating renal glomerular structure and function, we have produced an annotated transcriptome database for normal human glomeruli using the SAGE approach. Description: The database contains 22,907 unique SAGE tag sequences, with a total tag count of 48,905. For each SAGE tag, the ratio of its frequency in glomeruli relative to that in 115 non-glomerular tissues or cells, a measure of transcript enrichment in glomeruli, was calculated. A total of 133 SAGE tags representing well-characterized transcripts were enriched 10-fold or more in glomeruli compared to other tissues. Comparison of data from this study with a previous human glomerular Sau3A-anchored SAGE library reveals that 47 of the highly enriched transcripts are common to both libraries. Among these are the SAGE tags representing many podocyte-predominant transcripts like WT-1, podocin and synaptopodin. Enrichment of podocyte transcript tags SAGE library indicates that other SAGE tags observed at much higher frequencies in this glomerular compared to non-glomerular SAGE libraries are likely to be glomerulus-predominant. A higher level of mRNA expression for 19 transcripts represented by glomerulus-enriched SAGE tags was verified by RT-PCR comparing glomeruli to lung, liver and spleen. Conclusions: The database can be retrieved from, or interrogated online at http://cgap.nci.nih.gov/SAGE. The annotated database is also provided as an additional file with gene identification for 9,022, and matches to the human genome or transcript homologs in other species for 1,433 tags. It should be a useful tool for in silico mining of glomerular gene expression