HT-SIP: a semi-automated stable isotope probing pipeline identifies cross-kingdom interactions in the hyphosphere of arbuscular mycorrhizal fungi - Microbiome

DNA in each density fraction is then purified and concentrated using a Hamilton Microlab STAR liquid handling robot, which we have programmed to partially automate PEG precipitations using a previously published protocol [], with modifications for 96-well plates. We configured our robot deck to process four plates; this allows a maximum of 16 SIP samples to be processed simultaneously .

Finally, the DNA concentration of each fraction is quantified with a PicoGreen fluorescence assay . Picogreen quantification plates are prepared in triplicate on a Hamilton Microlab STAR robot, where each plate contains a row for the standard curve. Samples are mixed with the PicoGreen reagent in a 96-well intermediate mixing plate, and then distributed into three 96-well PCR plates for fluorescence analysis.

To compare automated versus manual PEG precipitations, 4 μg soil DNA was added per density gradient. Automated precipitations were performed as described above. For manual precipitations, PEG precipitations were conducted in microcentrifuge tubes as previously described [ ] using published centrifuge speeds and times, which we note are faster than those used for our HT-SIP plate-based method.Absorption of DNA to polypropylene tubes can lead to substantial sample loss, especially for DNA in high ionic strength solutions, but this concern can be mitigated by adding non-ionic detergents [].

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Vaginal microbiome-host interactions modeled in a human vagina-on-a-chip - MicrobiomeBackground A dominance of non-iners Lactobacillus species in the vaginal microbiome is optimal and strongly associated with gynecological and obstetric health, while the presence of diverse obligate or facultative anaerobic bacteria and a paucity in Lactobacillus species, similar to communities found in bacterial vaginosis (BV), is considered non-optimal and associated with adverse health outcomes. Various therapeutic strategies are being explored to modulate the composition of the vaginal microbiome; however, there is no human model that faithfully reproduces the vaginal epithelial microenvironment for preclinical validation of potential therapeutics or testing hypotheses about vaginal epithelium-microbiome interactions. Results Here, we describe an organ-on-a-chip (organ chip) microfluidic culture model of the human vaginal mucosa (vagina chip) that is lined by hormone-sensitive, primary vaginal epithelium interfaced with underlying stromal fibroblasts, which sustains a low physiological oxygen concentration in the epithelial lumen. We show that the Vagina Chip can be used to assess colonization by optimal L. crispatus consortia as well as non-optimal Gardnerella vaginalis-containing consortia, and to measure associated host innate immune responses. Co-culture and growth of the L. crispatus consortia on-chip was accompanied by maintenance of epithelial cell viability, accumulation of D- and L-lactic acid, maintenance of a physiologically relevant low pH, and down regulation of proinflammatory cytokines. In contrast, co-culture of G. vaginalis-containing consortia in the vagina chip resulted in epithelial cell injury, a rise in pH, and upregulation of proinflammatory cytokines. Conclusion This study demonstrates the potential of applying human organ chip technology to create a preclinical model of the human vaginal mucosa that can be used to better understand interactions between the vaginal microbiome and host tissues, as well as to evaluate the safety and efficac
Source: BioMedCentral - 🏆 22. / 71 Read more »

Exploring the effects of dietary fiber on gut microbiome and inflammatory diseasesExploring the effects of dietary fiber on gut microbiome and inflammatory diseases elsevierconnect Fiber DietaryFiber Gut GutMicrobiome InflammatoryDisease ElsevierConnect Good review of the importance of dietary fiber for microbial health!
Source: NewsMedical - 🏆 19. / 71 Read more »

Stones reserves special praise for Maguire after England star blocks out background noise - Football365John Stones is proud of the way England defensive partner Harry Maguire has drowned out the noise over his poor form to shine at the World Cup...
Source: F365 - 🏆 5. / 97 Read more »

Impact of the COVID-19 pandemic on timeliness and equity of measles, mumps and rubella vaccinations in North East London: a longitudinal study using electronic health recordsObjectives To quantify the effect of the COVID-19 pandemic on the timeliness of, and geographical and sociodemographic inequalities in, receipt of first measles, mumps and rubella (MMR) vaccination. Design Longitudinal study using primary care electronic health records. Setting 285 general practices in North East London. Participants Children born between 23 August 2017 and 22 September 2018 (pre-pandemic cohort) or between 23 March 2019 and 1 May 2020 (pandemic cohort). Main outcome measure Receipt of timely MMR vaccination between 12 and 18 months of age. Methods We used logistic regression to estimate the ORs (95% CIs) of receipt of a timely vaccination adjusting for sex, deprivation, ethnic background and Clinical Commissioning Group. We plotted choropleth maps of the proportion receiving timely vaccinations. Results Timely MMR receipt fell by 4.0% (95% CI: 3.4% to 4.6%) from 79.2% (78.8% to 79.6%) to 75.2% (74.7% to 75.7%) in the pre-pandemic ( n=33 226; 51.3% boys) and pandemic ( n=32 446; 51.4%) cohorts, respectively. After adjustment, timely vaccination was less likely in the pandemic cohort (0.79; 0.76 to 0.82), children from black (0.70; 0.65 to 0.76), mixed/other (0.77; 0.72 to 0.82) or with missing (0.77; 0.74 to 0.81) ethnic background, and more likely in girls (1.07; 1.03 to 1.11) and those from South Asian backgrounds (1.39; 1.30 to 1.48). Children living in the least deprived areas were more likely to receive a timely MMR (2.09; 1.78 to 2.46) but there was no interaction between cohorts and deprivation (Wald statistic: 3.44; p=0.49). The proportion of neighbourhoods where less than 60% of children received timely vaccination increased from 7.5% to 12.7% during the pandemic. Conclusions The COVID-19 pandemic was associated with a significant fall in timely MMR receipt and increased geographical clustering of measles susceptibility in an area of historically low and inequitable MMR coverage. Immediate action is needed to avert measles outbreaks and sup
Source: medical_xpress - 🏆 101. / 51 Read more »

USA vs Netherlands liveFollow coverage of the USMNT’s last-16 clash with Netherlands at the 2022 World Cup in Qatar live with The Athletic.The US are attempting to reach a World Cup quarter-final for the first time since 2002, while the Netherlands have reached three finals but never triumphed.The match will be played at the Khalifa International Stadium in Al-Rayyan and kicks off at 3pm GMT, 10am ET, 7am PT.United States: Matt Turner, Sergino Dest, Walker Zimmerman, Tyler Adams (capt.), Antonee Robinson, Yunus Musah, Weston McKennie, Christian Pulisic, Tim Ream, Tim Weah, Jesus FerreiraNetherlands: Andries Noppert, Daley Blind, Nathan Ake, Virgil van Dijk (capt.), Jurrien Timber, Denzel Dumfries, Marten de Roon, Davy Klaassen, Frenkie de Jong, Cody Gakpo, Memphis Depay.Background readingWhat does future hold for Pulisic?World Cup last 16 — who's playing and when?World Cup bracket — plot route to the final(Photo: Getty Images) Lets go 🇺🇸 !!!!! We are here to fck sht up !!!! Holland is next !!! KO 🥶🥶🥶🥶 ice cold Whomever is playing they have the heavy weight of a Countries Hopes and Dreams I'm no Tom Ranaldi so I can only say Fare Well USMNT
Source: TheAthletic - 🏆 54. / 63 Read more »