The reduction of acetylpyridine adenine dinucleotide by NADH: is it a significant reaction of proton-translocating transhydrogenase, or an artefact?

AbstractTranshydrogenase is a proton pump. It has separate binding sites for NAD+/NADH (on domain I of the protein) and for NADP+/NADPH (on domain III). Purified, detergent-dispersed transhydrogenase from Escherichia coli catalyses the reduction of the NAD+ analogue, acetylpyridine adenine dinucleotide (AcPdAD+), by NADH at a slow rate in the absence of added NADP+ or NADPH. Although it is slow, this reaction is surprizing, since transhydrogenase is generally thought to catalyse hydride transfer between NAD(H) – or its analogues and NADP(H) – or its analogues, by a ternary complex mechanism. It is shown that hydride transfer occurs between the 4A position on the nicotinamide ring of NADH and the 4A position of AcPdAD+. On the basis of the known stereospecificity of the enzyme, this eliminates the possibilities of transhydrogenation (a) from NADH in domain I to AcPdAD+ wrongly located in domain III; and (b) from NADH wrongly located in domain III to AcPdAD+ in domain I. In the presence of low concentrations of added NADP+ or NADPH, detergent-dispersed E. coli transhydrogenase catalyses the very rapid reduction of AcPdAD+ by NADH. This reaction is cyclic; it takes place via the alternate oxidation of NADPH by AcPdAD+ and the reduction of NADP+ by NADH, while the NADPH and NADP+ remain tightly bound to the enzyme. In the present work, it is shown that the rate of the cyclic reaction and the rate of reduction of AcPdAD+ by NADH in the absence of added NADP+/NADPH, have similar dependences on pH and on MgSO4 concentration and that they have a similar kinetic character. It is therefore suggested that the reduction of AcPdAD+ by NADH is actually a cyclic reaction operating, either with tightly bound NADP+/NADPH on a small fraction (<5%) of the enzyme, or with NAD+/NADH (or AcPdAD+/AcPdADH) unnaturally occluded within the domain III site. Transhydrogenase associated with membrane vesicles (chromatophores) of Rhodospirillum rubrum also catalyses the reduction of AcPdAD+ by NADH in the absence of added NADP+/NADPH. When the chromatophores were stripped of transhydrogenase domain I, that reaction was lost in parallel with `normal reverse' transhydrogenation (e.g., the reduction of AcPdAD+ by NADPH). The two reactions were fully recovered upon reconstitution with recombinant domain I protein. However, after repeated washing of the domain I-depleted chromatophores, reverse transhydrogenation activity (when assayed in the presence of domain I) was retained, whereas the reduction of AcPdAD+ by NADH declined in activity. Addition of low concentrations of NADP+ or NADPH always supported the same high rate of the NADH→AcPdAD+ reaction independently of how often the membranes were washed. It is concluded that, as with the purified E. coli enzyme, the reduction of AcPdAD+ by NADH in chromatophores is a cyclic reaction involving nucleotides that are tightly bound in the domain III site of transhydrogenase. However, in the case of R. rubrum membranes it can be shown with some certainty that the bound nucleotides are NADP+ or NADPH. The data are thus adequately explained without recourse to suggestions of multiple nucleotide-binding sites on transhydrogenase

Use of topical local anesthetics to control pain during treatment of hoof lesions in dairy cows

Research Areas: AgricultureFood Science & TechnologyABSTRACT - Hoof pathologies in dairy cows have a major effect on both production and animal welfare. Trimming of excess or diseased hoof tissue is essential for the treatment of many of these conditions. Trimming hoof lesions can cause severe pain, resulting in adverse behavioral responses with risk for animal and human safety. Interventions are usually carried out by nonveterinary technicians in the absence of pain management training. Pain control during trimming is not only an ethical obligation but also allows for better manipulation and more meticulous treatment. The aim of this study was to test the efficacy of Tri-Solfen (Bayer Australia Ltd., Pymble, NSW, Australia), a combination of local anesthetics in a topical gel form, containing lidocaine, bupivacaine, adrenaline, and cetrimide, for the treatment of pain associated with trimming of hoof lesions. Sixty-two Holstein-Frisian cows were selected for trimming at the drying-off period and were visually scored for lameness before entering the chute. After diagnosis of the hoof lesion but before deep trimming was initiated, each animal was randomly distributed to 2 groups: C, usual trimming with no pain control, and T, trimming with a local anesthetic formulation being applied immediately after live corium was exposed. During curative trimming, behavior observation was conducted by 2 observers blind to treatment. In 27 cows, algometry measurements were performed before and after the procedure to assess animal reaction to pressure. Lameness scoring was again performed as the cow left the chute. Nonparametric tests and ANOVA were performed. Results showed that use of the topical anesthetic formulation significantly reduced reaction to trimming and lameness score after trimming when compared with nontreated animals. Algometry values showed increased pressure threshold after application of topical anesthetics. This study suggests that the use of topical local anesthesia with lidocaine and bupivacaine helps reduce pain associated with corrective trimming of severe hoof lesions, enhancing animal welfare and potentially ensuring safety of trimmers.info:eu-repo/semantics/publishedVersio

Contact transmission of influenza virus between ferrets imposes a looser bottleneck than respiratory droplet transmission allowing propagation of antiviral resistance

Influenza viruses cause annual seasonal epidemics and occasional pandemics. It is important to elucidate the stringency of bottlenecks during transmission to shed light on mechanisms that underlie the evolution and propagation of antigenic drift, host range switching or drug resistance. The virus spreads between people by different routes, including through the air in droplets and aerosols, and by direct contact. By housing ferrets under different conditions, it is possible to mimic various routes of transmission. Here, we inoculated donor animals with a mixture of two viruses whose genomes differed by one or two reverse engineered synonymous mutations, and measured the transmission of the mixture to exposed sentinel animals. Transmission through the air imposed a tight bottleneck since most recipient animals became infected by only one virus. In contrast, a direct contact transmission chain propagated a mixture of viruses suggesting the dose transferred by this route was higher. From animals with a mixed infection of viruses that were resistant and sensitive to the antiviral drug oseltamivir, resistance was propagated through contact transmission but not by air. These data imply that transmission events with a looser bottleneck can propagate minority variants and may be an important route for influenza evolution

Developing the framework for a risk map for mite vectored viruses in wheat resulting from pre-harvest hail damage

There is a strong economic incentive to reduce mite-vectored virus outbreaks. Most outbreaks in the central High Plains of the United States occur in the presence of volunteer wheat that emerges before harvest as a result of hail storms. This study provides a conceptual framework for developing a risk map for wheat diseases caused by mite-vectored viruses based on pre-harvest hail events. Traditional methods that use NDVI were found to be unsuitable due to low chlorophyll content in wheat at harvest. Site-level hyperspectral reflectance from mechanically hailed wheat showed increased canopy albedo. Therefore, any increase in NIR combined with large increases in red reflectance near harvest can be used to assign some level of risk. The regional model presented in this study utilized Landsat TM/ETMþ data and MODIS imagery to help gap-fill missing data. NOAA hail maps that estimate hail size were used to refine the area most likely at risk. The date range for each year was shifted to account for annual variations in crop phenology based on USDA Agriculture statistics for percent harvest of wheat. Between 2003 and 2013, there was a moderate trend (R2 ¼ 0.72) between the county-level insurance claims for Cheyenne County, Nebraska and the area determined to be at risk by the model (excluding the NOAA hail size product due to limited availability) when years with low hail claims (\u3c400 ha) were excluded. These results demonstrate the potential of an operational risk map for mite-vectored viruses due to pre-season hail events

Developing the framework for a risk map for mite vectored viruses in wheat resulting from pre-harvest hail damage

There is a strong economic incentive to reduce mite-vectored virus outbreaks. Most outbreaks in the central High Plains of the United States occur in the presence of volunteer wheat that emerges before harvest as a result of hail storms. This study provides a conceptual framework for developing a risk map for wheat diseases caused by mite-vectored viruses based on pre-harvest hail events. Traditional methods that use NDVI were found to be unsuitable due to low chlorophyll content in wheat at harvest. Site-level hyperspectral reflectance from mechanically hailed wheat showed increased canopy albedo. Therefore, any increase in NIR combined with large increases in red reflectance near harvest can be used to assign some level of risk. The regional model presented in this study utilized Landsat TM/ETMþ data and MODIS imagery to help gap-fill missing data. NOAA hail maps that estimate hail size were used to refine the area most likely at risk. The date range for each year was shifted to account for annual variations in crop phenology based on USDA Agriculture statistics for percent harvest of wheat. Between 2003 and 2013, there was a moderate trend (R2 ¼ 0.72) between the county-level insurance claims for Cheyenne County, Nebraska and the area determined to be at risk by the model (excluding the NOAA hail size product due to limited availability) when years with low hail claims (\u3c400 ha) were excluded. These results demonstrate the potential of an operational risk map for mite-vectored viruses due to pre-season hail events

Citalopram Enhances the Activity of Chloroquine in Resistant Plasmodium in Vitro and in Vivo 1

ABSTRACT Citalopram, is an extremely potent inhibitor of neuronal serotonin reuptake. It is structurally unrelated to other antidepressants, but it contains the chemical features associated with reversal of drug resistance and exhibits minimal cardiotoxic side effects and fewer of the anticholinergic and adrenolytic side effects associated with other psychotropic agents. Sensitivity tests to citalopram alone and in combination with chloroquine were performed against chloroquine-resistant and chloroquine-sensitive strains of Plasmodium falciparum and Plasmodium chabaudi. Citalopram alone showed intrinsic activity against the chloroquine-resistant strains of P. falciparum (IC 50 ϭ 1.51 Ϯ .6 M) but only limited activity against the chloroquine-sensitive strain (IC 50 ϭ 33.27 Ϯ 5.87 M) and no activity in vivo. The interaction of chloroquine and citalopram in vitro resulted in a synergistic response in the chloroquineresistant strain but there was no interaction between the drugs in the chloroquine-sensitive strain-a pattern found with other reversal agents. Citalopram enhanced chloroquine susceptibility in both strains of P. chabaudi, however, the potentiating effect was seen at lower doses in the chloroquine-resistant strain. The results of this study suggest that citalopram may have potential as a chemosensitizer in Plasmodium infections on the basis of the low toxicity of citalopram at concentrations potentiating chloroquine activity both in vitro and in vivo. Malaria is a significant source of global morbidity and mortality. Despite the development of new antimalarial agents such as mefloquine, halofantrine and the artemisins, chloroquine remains the drug of choice for the treatment of uncomplicated Plasmodium falciparum malaria infections, due to its low cost, rapid onset of action and its low toxicity. However, the efficacy of chloroquine has diminished due to the emergence and prevalence of chloroquine-resistant strains of P. falciparum (Wensdorfer and Payne, 1991). The rapid development and spread of resistance to chloroquine and other antimalarials, and the tremendous cost of drug development has emphasized the necessity to optimize the use of existing antimalarial agents A number of adjunct drugs have been identified from a wide variety of chemical classes including calcium-channel blockers Citalopram,1-[3-(dimethylamino)propyl]-1-(4-fluorophenyl)-1,3dihydroisobenzofuran-5-carbonite), is an extremely potent inhibitor of neuronal serotonin reuptake The low toxicity coupled with the chemical similarity to chemosensitizers (resistance reversal agents) prompted us to investigate the chemosensitizing effect of citalopram in Plasmodium. In this study, we screened citalopram for chloroquine potentiating activity in chloroquine-resistant and chloroquine sensitive-parasites; both in vitro against P. falciparum and in a rodent malaria model (Plasmodium chabaudi). Methods Effect of Citalopram in Vitro Parasites. Two well-characterized isolates of P. falciparum were used for the drug assays. The chloroquine-resistant FCR-3 strain (IC 50 ϳ 150 nM) (donated by J. Freese, Research in diseases of the Tropical Environment, Durban, South Africa) and the chloroquine

Sharing best practices through online communities of practice: a case study

<p>Abstract</p> <p>Introduction</p> <p>The USAID-funded Capacity Project established the Global Alliance for Pre-Service Education (GAPS) to provide an online forum to discuss issues related to teaching and acquiring competence in family planning, with a focus on developing countries' health related training institutions. The success of the Global Alliance for Nursing and Midwifery's ongoing web-based community of practice (CoP) provided a strong example of the successful use of this medium to reach many participants in a range of settings.</p> <p>Case description</p> <p>GAPS functioned as a moderated set of forums that were analyzed by a small group of experts in family planning and pre-service education from three organizations. The cost of the program included the effort provided by the moderators and the time to administer responses and conduct the analysis.</p> <p>Discussion and evaluation</p> <p>Family planning is still considered a minor topic in health related training institutions. Rather than focusing solely on family planning competencies, GAPS members suggested a focus on several professional competencies (e.g. communication, leadership, cultural sensitivity, teamwork and problem solving) that would enhance the resulting health care graduate's ability to operate in a complex health environment. Resources to support competency-based education in the academic setting must be sufficient and appropriately distributed. Where clinical competencies are incorporated into pre-service education, responsible faculty and preceptors must be clinically proficient. The interdisciplinary GAPS memberships allowed for a comparison and contrast of competencies, opportunities, promising practices, documents, lessons learned and key teaching strategies.</p> <p>Conclusions</p> <p>Online CoPs are a useful interface for connecting developing country experiences. From CoPs, we may uncover challenges and opportunities that are faced in the absorption of key public health competencies required for decreasing maternal mortality and morbidity. Use of the World Health Organization (WHO) Implementing Best Practices Knowledge Gateway, which requires only a low bandwidth connection, gave educators an opportunity to engage in the discussion even in the most Internet access-restricted places (e.g. Ethiopia). In order to sustain an online CoP, funds must come from an international organization (e.g. WHO regional office) or university that can program the costs long-term. Eventually, the long-term effectiveness and sustainability of GAPS rests on its transfer to the members themselves.</p

The cost of health professionals' brain drain in Kenya

BACKGROUND: Past attempts to estimate the cost of migration were limited to education costs only and did not include the lost returns from investment. The objectives of this study were: (i) to estimate the financial cost of emigration of Kenyan doctors to the United Kingdom (UK) and the United States of America (USA); (ii) to estimate the financial cost of emigration of nurses to seven OECD countries (Canada, Denmark, Finland, Ireland, Portugal, UK, USA); and (iii) to describe other losses from brain drain. METHODS: The costs of primary, secondary, medical and nursing schools were estimated in 2005. The cost information used in this study was obtained from one non-profit primary and secondary school and one public university in Kenya. The cost estimates represent unsubsidized cost. The loss incurred by Kenya through emigration was obtained by compounding the cost of educating a medical doctor and a nurse over the period between the average age of emigration (30 years) and the age of retirement (62 years) in recipient countries. RESULTS: The total cost of educating a single medical doctor from primary school to university is US$65,997; and for every doctor who emigrates, a country loses about US$ 517,931 worth of returns from investment. The total cost of educating one nurse from primary school to college of health sciences is US$43,180; and for every nurse that emigrates, a country loses about US$ 338,868 worth of returns from investment. CONCLUSION: Developed countries continue to deprive Kenya of millions of dollars worth of investments embodied in her human resources for health. If the current trend of poaching of scarce human resources for health (and other professionals) from Kenya is not curtailed, the chances of achieving the Millennium Development Goals would remain bleak. Such continued plunder of investments embodied in human resources contributes to further underdevelopment of Kenya and to keeping a majority of her people in the vicious circle of ill-health and poverty. Therefore, both developed and developing countries need to urgently develop and implement strategies for addressing the health human resource crisis

Extensive Sheep and Goat Production: The Role of Novel Technologies towards Sustainability and Animal Welfare

ReviewSheep and goat extensive production systems are very important in the context of global food security and the use of rangelands that have no alternative agricultural use. In such systems, there are enormous challenges to address. These include, for instance, classical production issues, such as nutrition or reproduction, as well as carbon-efficient systems within the climate-change context. An adequate response to these issues is determinant to economic and environmental sustainability. The answers to such problems need to combine efficiently not only the classical production aspects, but also the increasingly important health, welfare, and environmental aspects in an integrated fashion. The purpose of the study was to review the application of technological developments, in addition to remote-sensing in tandem with other state-of-the-art techniques that could be used within the framework of extensive production systems of sheep and goats and their impact on nutrition, production, and ultimately, the welfare of these species. In addition to precision livestock farming (PLF), these include other relevant technologies, namely omics and other areas of relevance in small-ruminant extensive production: heat stress, colostrum intake, passive immunity, newborn survival, biomarkers of metabolic disease diagnosis, and parasite resistance breeding. This work shows the substantial, dynamic nature of the scientific community to contribute to solutions that make extensive production systems of sheep and goats more sustainable, efficient, and aligned with current concerns with the environment and welfareinfo:eu-repo/semantics/publishedVersio

Changes in the Management of Patients having Radical Radiotherapy for Lung Cancer during the First Wave of the COVID-19 Pandemic in the UK.

AIMS: In response to the COVID-19 pandemic, guidelines on reduced fractionation for patients treated with curative-intent radiotherapy were published, aimed at reducing the number of hospital attendances and potential exposure of vulnerable patients to minimise the risk of COVID-19 infection. We describe the changes that took place in the management of patients with stage I-III lung cancer from April to October 2020. MATERIALS AND METHODS: Lung Radiotherapy during the COVID-19 Pandemic (COVID-RT Lung) is a prospective multicentre UK cohort study. The inclusion criteria were: patients with stage I-III lung cancer referred for and/or treated with radical radiotherapy between 2nd April and 2nd October 2020. Patients who had had a change in their management and those who continued with standard management were included. Data on demographics, COVID-19 diagnosis, diagnostic work-up, radiotherapy and systemic treatment were collected and reported as counts and percentages. Patient characteristics associated with a change in treatment were analysed using multivariable binary logistic regression. RESULTS: In total, 1553 patients were included (median age 72 years, 49% female); 93 (12%) had a change to their diagnostic investigation and 528 (34%) had a change to their treatment from their centre's standard of care as a result of the COVID-19 pandemic. Age ≥70 years, male gender and stage III disease were associated with a change in treatment on multivariable analysis. Patients who had their treatment changed had a median of 15 fractions of radiotherapy compared with a median of 20 fractions in those who did not have their treatment changed. Low rates of COVID-19 infection were seen during or after radiotherapy, with only 21 patients (1.4%) developing the disease. CONCLUSIONS: The COVID-19 pandemic resulted in changes to patient treatment in line with national recommendations. The main change was an increase in hypofractionation. Further work is ongoing to analyse the impact of these changes on patient outcomes