Distinct NAD(P)H binding configurations revealed by fluorescence lifetime, anisotropy and polarised two-photon absorption

NADH and NADPH play key roles in the regulation of metabolism. Their endogenous fluorescence is sensitive to enzyme binding, allowing changes in cellular metabolic state to be determined using fluorescence lifetime imaging microscopy (FLIM). However, to fully uncover the underlying biochemistry, the relationships between their fluorescence and binding dynamics require greater understanding. Here we accomplish this through time- and polarisation-resolved fluorescence and polarised two-photon absorption measurements. Two lifetimes result from binding of both NADH to lactate dehydrogenase and NADPH to isocitrate dehydrogenase. The composite fluorescence anisotropy indicates the shorter (1.3-1.6ns) decay component to be accompanied by local motion of the nicotinamide ring, pointing to attachment solely via the adenine moiety. For the longer lifetime (3.2-4.4ns), the nicotinamide conformational freedom is found to be fully restricted. As full and partial nicotinamide binding are recognised steps in dehydrogenase catalysis, our results unify photophysical, structural and functional aspects of NADH and NADPH binding and clarify the biochemical processes that underlie their contrasting intracellular lifetimes

Flooding in Southwestern Pennsylvania: Knowledge Gaps and Approaches

As southwestern Pennsylvania continues to evolve from its industrial past to become a national leader in innovation and sustainability, it has encountered a complex set of water challenges that threaten the economy, ecology, and public health of the region. In this context, over the last year, the Heinz Endowments funded the Pittsburgh Collaboratory for Water Research, Education, and Outreach at the University of Pittsburgh to hold a series of consensus-building meetings among regional academic scholars, community groups, governmental, and non-governmental organizations. These meetings (one each on green infrastructure, water quality, and flooding) aimed to identify key regional knowledge gaps and chart a collaborative research agenda to fill these gaps and enhance the region’s ability to strategically and creatively solve water problems. In June 2019, the first of the reports on Green Infrastructure and Stormwater Management was released. In October 2019, the second report on Water Quality was released. Now, in February 2020, the third report describing the research agenda on flooding arising from an April 2, 2019 meeting is being formally released. This report outlines several fundamental knowledge gaps in the region and suggests methods to span these gaps with new collaborative research

Influence of prepartum nutrition on cow performance and subsequent calf performance, glucose tolerance, carcass characteristics, efficiency, and gene expression

Previous research has demonstrated the potential for cow nutrition during mid- and late gestation to have lasting effects on progeny growth, carcass characteristics, and insulin sensitivity. With the advancement of nutrigenomics, the ability to evaluate the effects of maternal nutrition on gene expression in tissues of subsequent progeny has grown. The objective of this dissertation was to evaluate potential fetal programming effects of beef cow nutrition within beef production systems. Fall-calving cows grazing endophyte-fescue experience a decline in forage quality during late gestation, when fetal growth occurs at a high rate. Fall-calving, mature cows and their progeny were used to evaluate the effects of late gestation dried distillers grains plus solubles (DDGS) supplementation on cow performance, and progeny growth and carcass characteristics. Cows were offered daily DDGS supplementation or offered no supplement 69 ± 9 d prepartum through calving. Cow BW and BCS change during late gestation were greater (P ≤ 0.02) for cows offered supplement. Supplementation had no effect (P ≥ 0.11) on calving date, calf birth or weaning BW, or pre-weaning ADG, milk production, AI conception, or overall pregnancy rate. For steer progeny, dam DDGS supplementation had no effect (P ≥ 0.19) on feedlot performance or carcass characteristics. Use of corn coproducts in drylot beef gestation rations has increased over the last decade. Because of the elevated energy and CP content of corn coproducts, cow drylot rations may easily exceed cow energy and CP requirements. The effects of excessive prepartum dietary energy and CP intake of subsequent progeny needs to be further investigated. In one experiment, spring-calving, mature cows and their progeny were used to evaluate the effects of prepartum dietary energy on cow performance as well as performance and carcass characteristics of subsequent progeny. Cows were limit-fed isonitrogenous rations that provided 100% or 125% of TDN requirements, respectively, from 83 ± 10 d prepartum to calving. Cow BW change through breeding was greater (P < 0.01) and BCS change tended to be greater (P = 0.09) for cows 125% of TDN requirement. Birth BW was greater (P = 0.02) for calves born to cows fed excessive dietary energy with no increase (P = 0.30) in percentage of unassisted births. There were no effects (P ≥ 0.27) of dietary energy on calving date, milk production, or subsequent pregnancy rate. Calf feedlot performance was not affected (P ≥ 0.20) by prepartum energy. Although progeny born to dams fed excessive dietary energy tended (P = 0.10) to have greater marbling scores at weaning, prepartum dietary energy had no effect (P ≥ 0.60) on carcass marbling score or other carcass characteristics. Two experiments were conducted to investigate the effects of prepartum dietary CP intake on cow performance as well as subsequent progeny growth, carcass characteristics, and plasma glucose and insulin concentrations. Spring-calving, mature cows in both experiments were limit-fed rations formulated to be isocaloric, and provide 100% or 129% of CP requirement, respectively, from 92 ± 10 or 78 ± 12 d prepartum to calving in Exp. 1 and 2, respectively. Prepartum CP intake had no effect on cow BW, BCS, milk production, and subsequent reproduction or progeny pre-weaning growth in either experiment. In Exp. 1, post-weaning ADG, final BW, and HCW were decreased in progeny born to dams fed excessive dietary CP. In Exp. 2, progeny post-weaning growth was not affected by treatment; yet, 12th rib fat thickness, KPH, and yield grade were greater for progeny born to dams fed excessive dietary CP. Progeny born to dams fed excessive dietary CP had decreased glucose and insulin concentrations, and insulin to glucose ratios, indicating greater insulin sensitivity. Mid-gestation is a critical time for the development of skeletal muscle and adipogenesis, two tissues of great interest to the beef industry. Much of the work that has demonstrated promising fetal programming effects, was done in nutrient restricted or overfed dams, 50% or 150% or energy and protein requirements, respectively. One experiment was conducted to investigate the effects of divergent plane of nutrition during mid-gestation on cow performance and progeny growth, feed efficiency, methane production, glucose tolerance, carcass characteristics, and gene expression in LM. Mature cows were limit-fed 3 planes of nutrition from 196 ± 14 to 114 ± 14 d prepartum: 100% energy and protein requirement (REQ), 70% requirement (70%REQ), or 130% requirement (130%REQ). Cow ADG during mid-gestation was greatest (P < 0.01) for 130%REQ, least for 70%REQ, with REQ intermediate. Transition period (30 d post-weaning) ADG was greatest (P < 0.01) for 70%REQ progeny, least for REQ progeny, with 130%REQ progeny intermediate. Treatment had no effect (P ≥ 0.21) on progeny RFI or 24 h methane emissions; but, REQ progeny tended (P = 0.06) to have decreased RG relative to 70%REQ and 130%REQ progeny. Treatment had no effect (P ≥ 0.12) on progeny glucose and insulin concentrations, AUC, glucose clearance rate, or insulin to glucose ratio during glucose tolerance test. Progeny of REQ dams had greater (P = 0.04) HCW than 70%REQ progeny, with 130%REQ intermediate and not different from either. Treatment had no effect (P ≥ 0.27) on other carcass characteristics. Treatment had no effect (P ≥ 0.24) on expression of 13 selected genes in LM, with the exception of MYH1 (P = 0.07). Expression of all target genes, except MYH7, changed (P ≤ 0.03) as progeny d of age increased. There was a treatment by d of age interaction (P = 0.04) for expression of MYH1, in which expression of MYH1 was least in progeny born to 70%REQ dams at 392 ± 14 d of age relative to REQ and 130% progeny. There were treatment by progeny sex by d of age interactions (P ≤ 0.01) for expression of SLC2A4, YY1, and MYH7. In conclusion, prepartum DDGS supplementation improved cow BW and BCS but did not alter milk production, subsequent reproduction, or subsequent calf performance or carcass characteristics. Feeding cows 125% of TDN requirement during late gestation increased cow BW change and progeny birth body weight; but, had no effect on pregnancy rate or progeny performance. Although feeding cows 129% of CP requirement during late gestation did not affect cow performance, progeny post-weaning growth was decreased and carcass adiposity was increased. Differing mid-gestation plane of nutrition diverged cow BW and affected transition period ADG and HCW of progeny; yet, did not dramatically impact methane production, insulin resistance, or gene expression in LM

Disrupted Maturation of the Microbiota and Metabolome among Extremely Preterm Infants with Postnatal Growth Failure

Growth failure during infancy is a major global problem that has adverse effects on long-term health and neurodevelopment. Preterm infants are disproportionately affected by growth failure and its effects. Herein we found that extremely preterm infants with postnatal growth failure have disrupted maturation of the intestinal microbiota, characterized by persistently low diversity, dominance of pathogenic bacteria within the Enterobacteriaceae family, and a paucity of strictly anaerobic taxa including Veillonella relative to infants with appropriate postnatal growth. Metabolomic profiling of infants with growth failure demonstrated elevated serum acylcarnitines, fatty acids, and other byproducts of lipolysis and fatty acid oxidation. Machine learning algorithms for normal maturation of the microbiota and metabolome among infants with appropriate growth revealed a pattern of delayed maturation of the microbiota and metabolome among infants with growth failure. Collectively, we identified novel microbial and metabolic features of growth failure in preterm infants and potentially modifiable targets for intervention

Identifying Candida albicans Gene Networks Involved in Pathogenicity

Acknowledgments This is a short text to acknowledge the contributions of specific colleagues, institutions, or agencies that aided the efforts of the authors. Funding RA was generously supported by a Wellcome Trust Institutional Strategic Support Award [WT105618MA], a Microbiology Research Visit Grant [RVG16/18], and a EPSRC/BBSRC Innovation Fellowship [EP/S001352/1]. AB was supported by a programme grant from the UK Medical Research Council [MR/M026663/1] and by the Medical Research Council Centre for Medical Mycology at the University of Aberdeen [MR/N006364/1]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewedPublisher PD

Knowledge and Awareness of HPV Vaccine and Acceptability to Vaccinate in Sub-Saharan Africa: A Systematic Review

Objectives: We assessed the knowledge and awareness of cervical cancer, HPV and HPV vaccine, and willingness and acceptability to vaccinate in sub-Saharan African (SSA) countries. We further identified countries that fulfill the two GAVI Alliance eligibility criteria to support nationwide HPV vaccination. Methods: We conducted a systematic review of peer-reviewed studies on the knowledge and awareness of cervical cancer, HPV and HPV vaccine, and willingness and acceptability to vaccinate. Trends in Diphtheria-tetanus-pertussis (DTP3) vaccine coverage in SSA countries from 1990–2011 were extracted from the World Health Organization database. Findings: The review revealed high levels of willingness and acceptability of HPV vaccine but low levels of knowledge and awareness of cervical cancer, HPV or HPV vaccine. We identified only six countries to have met the two GAVI Alliance requirements for supporting introduction of HPV vaccine: 1) the ability to deliver multi-dose vaccines for no less than 50% of the target vaccination cohort in an average size district, and 2) achieving over 70% coverage of DTP3 vaccine nationally. From 2008 through 2011 all SSA countries, with the exception of Mauritania and Nigeria, have reached or maintained DTP3 coverage at 70% or above. Conclusion: There is an urgent need for more education to inform the public about HPV, HPV vaccine, and cervical cancer, particularly to key demographics, (adolescents, parents and healthcare professionals), to leverage high levels of willingness and acceptability of HPV vaccine towards successful implementation of HPV vaccination programs. There is unpreparedness in most SSA countries to roll out national HPV vaccination as per the GAVI Alliance eligibility criteria for supporting introduction of the vaccine. In countries that have met 70% DTP3 coverage, pilot programs need to be rolled out to identify the best practice and strategies for delivering HPV vaccines to adolescents and also to qualify for GAVI Alliance support