GTP hydrolysis by complexes of the signal recognition particle and the signal recognition particle receptor

Translocation of proteins across the endoplasmic reticulum membrane is a GTP-dependent process. The signal recognition particle (SRP) and the SRP receptor both contain subunits with GTP binding domains. One GTP-dependent reaction during protein translocation is the SRP receptor-mediated dissociation of SRP from the signal sequence of a nascent polypeptide. Here, we have assayed the SRP and the SRP receptor for GTP binding and hydrolysis activities. GTP hydrolysis by SRP was not detected, so the maximal GTP hydrolysis rate for SRP was estimated to be \u3c 0.002 mol GTP hydrolyzed x mol of SRP-1 x min-1. The intrinsic GTP hydrolysis activity of the SRP receptor ranged between 0.02 and 0.04 mol GTP hydrolyzed x mol of SRP receptor-1 x min-1. A 40-fold enhancement of GTP hydrolysis activity relative to that observed for the SRP receptor alone was obtained when complexes were formed between SRP and the SRP receptor. GTP hydrolysis activity was inhibited by GDP, but not by ATP. Extended incubation of the SRP or the SRP receptor with GTP resulted in substoichiometric quantities of protein-bound ribonucleotide. SRP-SRP receptor complexes engaged in GTP hydrolysis were found to contain a minimum of one bound guanine ribonucleotide per SRP-SRP receptor complex. We conclude that the GTP hydrolysis activity described here is indicative of one of the GTPase cycles that occur during protein translocation across the endoplasmic reticulum

Access of proteinase K to partially translocated nascent polypeptides in intact and detergent-solubilized membranes

We have used proteinase K as a probe to detect cytoplasmically and luminally exposed segments of nascent polypeptides undergoing transport across mammalian microsomal membranes. A series of translocation intermediates consisting of discrete-sized nascent chains was prepared by including microsomal membranes in cell-free translations of mRNAs lacking termination codons. The truncated mRNAs were derived from preprolactin and the G protein of vesicular stomatitis virus and encoded nascent chains ranging between 64 and 200 amino acid residues long. Partially translocated nascent chains of 100 amino acid residues or less were insensitive to protease digestion from the external surface of the membrane while longer nascent chains were susceptible to digestion by externally added protease. We conclude that the increased protease sensitivity of larger nascent chains is due to the exposure of a segment of the nascent polypeptide on the cytoplasmic face of the membrane. In contrast, low molecular weight nascent chains were remarkably resistant to protease digestion even after detergent solubilization of the membrane. The protease resistant behaviour of detergent solubilized nascent chains could be abolished by release of the polypeptide from the ribosome or by the addition of protein denaturants. We propose that the protease resistance of partially translocated nascent chains can be ascribed to components of the translocation apparatus that remain bound to the nascent chain after detergent solubilization of the membrane

Effects of Seasonal Variation in Milk Composition on the Quality of Pizza Cheese

End of Project ReportThe main aims of this study were to investigate the effects of diet and lactation stage on the composition and cheesemaking quality of milk produced under controlled conditions. The main conclusions were as follows: These studies clearly demonstrated that the Recommended Moorepark Milk Production System in conjunction with an objectively standardised cheesemaking process provides a model for year round production of quality Mozzarella cheese. Databases have been established on the effects of diet quantity and quality, and stage of lactation on the composition, processability and cheesemaking characteristics of milk from both Spring- and Autumn-calving herds. Increasing the daily herbage allowance from 16 to 24 kgs DM/cow/day during mid-lactation, resulted in increases in the level of milk casein and cheese yield but had little influence on cheese functionality. Similarily improving diet quality in mid-lactation by reducing the stocking density from 4.3 to 3.8 cows/ha combined with concentrate supplementation (3 kgs/cow/day) had the same effect. Using milk from a Spring-calving herd, produced according to the Recommended Moorepark Milk Production System, in conjunction with an objectively-standardised Mozzarella cheesemaking process, no major problems were encountered during the lactation period 170 - 273 days from calving. Extending the lactation period of the Spring-calving herd from ~ 273 to 286 days resulted in higher cheese moisture (by ~ 2%), softer cheese, and lower chewiness in the melted cheese. A sharp decline in both total protein, casein and lactose in the milk was observed during this period. However the blending of this milk with milk of an Autumn-calving herd overcame these cheesemaking problems. The yield of low moisture Mozzarella cheese (using milks from Spring- or Autumn-calved herds) was positively correlated with milk casein level. The yield of cheese from the Spring-calved herd increased concomitantly with increasing casein level to day 273 of lactation and decreased thereafter as the casein concentration declined. In these studies it was found that easy-to-use tests such as lactose level in the milk and rennet coagulation properties as determined by Formagraph were useful indicators of the suitability of milk for cheesemaking. The Recommended Moorepark Milk Production System, as applied in the late lactation period, was characterised by a high plane of nutrition and a drying-off strategy which ensured a minimum daily milk yield per cow of 9 kg. It resulted in milk of good cheesemaking quality - lactose > 4.25%, and casein > 2.6% and satisfactory rennet coagulation properties - curd firming rate of > 0.15 min ¯¹ curd firmness at 60 min of > 45mm - at the end of lactation.Department of Agriculture, Food and the Marin

ASPIRE-2-PREVENT: a survey of lifestyle, risk factor management and cardioprotective medication in patients with coronary heart disease and people at high risk of developing cardiovascular disease in the UK.

OBJECTIVE: To determine in patients with coronary heart disease (CHD) and people at high risk of developing cardiovascular disease (CVD) whether the Joint British Societies' guidelines on CVD prevention (JBS2) are followed in everyday clinical practice. DESIGN: A cross-sectional survey was undertaken of medical records and patient interviews and examinations at least 6 months after the recruiting event or diagnosis using standardised instruments and a central laboratory for measurement of lipids and glucose. SETTINGS: The ASPIRE-2-PREVENT survey was undertaken in 19 randomly selected hospitals and 19 randomly selected general practices in 12 geographical regions in England, Northern Ireland, Wales and Scotland. PATIENTS: In hospitals, 1474 consecutive patients with CHD were identified and 676 (25.6% women) were interviewed. In general practice, 943 people at high CVD risk were identified and 446 (46.5% women) were interviewed. RESULTS: The prevalence of risk factors in patients with CHD and high-risk individuals was, respectively: smoking 14.1%, 13.3%; obesity 38%, 50.2%; not reaching physical activity target 83.3%, 85.4%; blood pressure ≥130/80 mm Hg (patients with CHD and self-reported diabetes) or ≥140/85 mm Hg (high-risk individuals) 46.9%, 51.3%; total cholesterol ≥4 mmol/l 52.6%, 78.7%; and diabetes 17.8%, 43.8%. CONCLUSIONS: The potential among patients with CHD and individuals at high risk of developing CVD in the UK to achieve the JBS2 lifestyle and risk factor targets is considerable. CVD prevention needs a comprehensive multidisciplinary approach, addressing all aspects of lifestyle and risk factor management. The challenge is to engage and motivate cardiologists, physicians and other health professionals to routinely practice high quality preventive cardiology in a healthcare system which must invest in prevention

Leveraging Implementation Science to Understand Factors Influencing Sustained Use of Mental Health Apps: a Narrative Review

Mental health (MH) smartphone applications (apps), which can aid in self-management of conditions such as depression and anxiety, have demonstrated dramatic growth over the past decade. However, their effectiveness and potential for sustained use remain uncertain. This narrative review leverages implementation science theory to explore factors influencing MH app uptake. The review is guided by the integrated Promoting Action on Research Implementation in Health Services (i-PARIHS) framework and discusses the role of the innovation, its recipients, context, and facilitation in influencing successful implementation of MH apps. The review highlights critical literature published between 2015 and 2020 with a focus on depression and anxiety apps. Sources were identified via PubMed, Google Scholar, and Twitter using a range of keywords pertaining to MH apps. Findings suggest that for apps to be successful, they must be advantageous over alternative tools, relatively easy to navigate, and aligned with users\u27 needs, skills, and resources. Significantly more attention must be paid to the complex contexts in which MH app implementation is occurring in order to refine facilitation strategies. The evidence base is still uncertain regarding the effectiveness and usability of MH apps, and much can be learned from the apps we use daily; namely, simpler is better and plans to integrate full behavioral treatments into smartphone form may be misguided. Non-traditional funding mechanisms that are nimble, responsive, and encouraging of industry partnerships will be necessary to move the course of MH app development in the right direction

Genetic Variants of Milk Proteins - Relevance to Milk Composition and Cheese Production.

End of Project ReportObjectives: (i) to develop rapid screening procedures for the determination of milk protein polymorphism (genetic variants) (ii) to determine the frequency distribution of milk protein genetic variants in a large population of Irish Holstein-Friesians and to determine if there was an association between κ-casein variant and milk yield and composition in this group of animals, and (iii) to make Cheddar and low-moisture part-skim Mozzarella cheese from different κ-casein genetic variant milks and to assess any effect on cheese yield, composition and functional characteristics. Conclusions:Analysis of 6,007 individual Irish Holstein-Friesian milks showed that the phenotype distribution of the κ-casein BB variant was very low at 1.98% compared to 53.07% for κ-casein AA and 44.95% for κ-casein AB. While no statistically significant associations were observed between κ-casein variant and milk yield and composition, κ-casein BB variant milks had superior rennet coagulation properties to that of the AA or AB variants. Generally, κ-casein variant had little effect on compositional attributes of cheese apart from FDM (fat in dry matter) which was significantly higher in cheeses from κ-casein BB milk than in those from κ-casein AA milk. Generally, κ-casein variant had no significant effects on either primary or secondary proteolysis, or on the sensory and/or textural characteristics of Cheddar or Mozzarella cheese throughout ripening; or on the functional characteristics (e.g. flow and stretch) of baked Mozzarella on storage for 90 days at 4°C. However, κ-casein BB variant milk gave significantly higher actual, and moisture adjusted yields of Cheddar and Mozzarella cheese than either κ-casein AB or AA variant milks. For example, the moisture adjusted Cheddar yield from κ-casein BB milk was 8.2% higher than from κ-casein AA milk. In the case of Mozzarella, the moisture adjusted yield was 12% higher. Based on the results, it is estimated that the actual yield of cheese in a plant producing 20,000 tonnes per year from κ-casein AA milk would increase to approximately 21,180 tonnes of Cheddar, or 21,780 tonnes of Mozzarella if made from κ-casein BB milk. Where κ-casein AB milk is used instead of κ-casein BB milk, the estimated yield of Mozzarella would increase to 21,580 tonnes.Department of Agriculture, Food and the Marin

Effect of smoking on comparative efficacy of antiplatelet agents: systematic review, meta-analysis, and indirect comparison

Objective: To evaluate whether smoking status is associated with the efficacy of antiplatelet treatment in the prevention of cardiovascular events. Design: Systematic review, meta-analysis, and indirect comparisons. Data sources Medline (1966 to present) and Embase (1974 to present), with supplementary searches in databases of abstracts from major cardiology conferences, the Cumulative Index to Nursing and Allied Health (CINAHL) and the CAB Abstracts databases, and Google Scholar. Study selection Randomized trials of clopidogrel, prasugrel, or ticagrelor that examined clinical outcomes among subgroups of smokers and nonsmokers. Data extraction Two authors independently extracted all data, including information on the patient populations included in the trials, treatment types and doses, definitions of clinical outcomes and duration of follow-up, definitions of smoking subgroups and number of patients in each group, and effect estimates and 95% confidence intervals for each smoking status subgroup. Results: Of nine eligible randomized trials, one investigated clopidogrel compared with aspirin, four investigated clopidogrel plus aspirin compared with aspirin alone, and one investigated double dose compared with standard dose clopidogrel; these trials include 74 489 patients, of whom 21 717 (29%) were smokers. Among smokers, patients randomized to clopidogrel experienced a 25% reduction in the primary composite clinical outcome of cardiovascular death, myocardial infarction, and stroke compared with patients in the control groups (relative risk 0.75, 95% confidence interval 0.67 to 0.83). In nonsmokers, however, clopidogrel produced just an 8% reduction in the composite outcome (0.92, 0.87 to 0.98). Two studies investigated prasugrel plus aspirin compared with clopidogrel plus aspirin, and one study investigated ticagrelor plus aspirin compared with clopidogrel plus aspirin. In smokers, the relative risk was 0.71 (0.61 to 0.82) for prasugrel compared with clopidogrel and 0.83 (0.68 to 1.00) for ticagrelor compared with clopidogrel. Corresponding relative risks were 0.92 (0.83 to 1.01) and 0.89 (0.79 to 1.00) among nonsmokers. Conclusions: In randomized clinical trials of antiplatelet drugs, the reported clinical benefit of clopidogrel in reducing cardiovascular death, myocardial infarction, and stroke was seen primarily in smokers, with little benefit in nonsmokers

Defining a Paradigm for the Dissemination of Health Information to Immigrant Populations at the Fletcher Free Library

Abstract: In recent years, the surge of refugee families to the greater Burlington area has lead to a significant increase in the minority population of the city. The Fletcher Free Library (FFL) represents a potential health information resource for this population; it is, however, little utilized. Our project sought to target one of these minority populations, the Somali Bantu, and to diminish the barriers to the use of the FFL’s resources within the Somali population itself. To reduce barriers to access of health information by the Somali Bantu population, we educated the FFl’s reference librarians on Somali culture, developed a compendium of health information in both English and Somali for inclusion within the library’s collection, and staged a one-day intervention at the Community Health Center of Burlington to present the FFL as a potential source of health information for refugee populations. Somali Bantu use of the library, as well as reference librarian confidence in serving this minority population,was objectively assessed via pre- and post-interventional surveys.https://scholarworks.uvm.edu/comphp_gallery/1022/thumbnail.jp