A Simple and Reliable Reporter Assay to Test siRNA-Mediated Silencing of HIV-1 Gene Functions

The human immunodeficiency virus (HIV-1) infects and kills CD4+ T-lymphocytes causing a progressive loss of host immune competence, which ultimately leads to AIDS. RNA interference, as mediated by short interfering RNAs (siRNAs) designed to target viral mRNAs and expressed endogenously, offer a potential gene therapy approach to inhibit HIV replication. However, a simple and reliable method to screen the silencing activity of particular anti-HIV siRNAs is useful prior to conducting more extensive experimentation to determine the downstream effects on viral replication. Here, a short hairpin RNA (shRNA) targeting HIV-1 Rev (Revsh8526) was designed and cloned into an expression plasmid under the control of the RNA Polymerase III H1 promoter. To test the ability of Revsh8526 to silence Rev, a chimeric β-galactosidase reporter plasmid containing Rev exon 2 (β-galactosidase-RevE2) was generated. The ability of Revsh8526 to target and inhibit reporter gene expression was shown by X-gal staining and ortho-nitrophenyl-β-galactoside (ONPG) assay. The results of these assays indicated Revsh8526 effectively targeted and significantly inhibited β-galactosidase expression as compared to a non-targeted shRNA. These results show the utility of the chimeric β-galactosidase reporter in initial screening of siRNA reagents

Trigger point dry needling for musculoskeletal pain and disability: a systematic review of comparative effectiveness research

Background: Dry needling (DN) has been proposed to reduce pain and improve function related to myofascial trigger points (MTrPs). Several primary studies and systematic reviews have been conducted to examine the effect of DN versus placebo. However the comparative effectiveness of DN and established interventions has yet to be established. Purpose: The purpose of this systematic review was to determine whether DN was more effective than other established therapies to treat MTrPs. Data Sources: MEDLINE Complete, EBSCO, CINAHL, Sport Discus and Cochrane library databases were searched. Study Selection: Randomized controlled trials that used DN directed to MTrPs and used at least one other intervention method were included. Studies that had a placebo or sham group were excluded. Data Extraction: Of 394 records screened, 8 studies met the established criteria. The quality of each study was assessed using the PEDro scale. Data Synthesis: When DN was compared to standard therapy programs, 3 of the 4 studies found that DN was more effective in reducing pain and 1 found no difference. When DN was compared to stretching, DN reduced pain more effectively. Dry needling was not significantly more effective than high-power pain threshold ultrasound (US), laser, non steroidal anti inflammatory drugs, and percutaneous electrical nerve stimulation (PENS). Limitations: Included studies were relatively small and some lacked sound methodology. Conclusions: The results are mixed on the effectiveness of DN over standard rehab. More large scale, high quality studies are needed before definitive decisions can be made about the role of DN in physical therapy practice

Effects of epicatechin on cardiovascular function in middle-Aged diet-induced obese rat models of metabolic syndrome

The current study aimed to investigate the cardiovascular effects of epicatechin, a flavonoid found in green tea and cocoa, in attenuating complications associated with metabolic syndrome in diet-induced obese rats. Male Wistar-Kyoto (WKY) rats aged 16 weeks were fed either standard rat chow or given a high-fat-high-carbohydrate (HFHC) diet for 20 weeks. Epicatechin treatment (5 mg/kg/d) was administered to a subset of WKY rats commencing at week 8 of the 20 week HFHC feeding period. Body weights, food, water and energy intakes, blood pressure, heart rate and glucose tolerance were measured throughout the treatment period. Oxidative stress and inflammatory markers, lipid levels, cardiac collagen deposition, cardiac electrical function, aortic and mesenteric vessel reactivity were examined after the treatment. Twenty weeks of HFHC feeding in WKY rats resulted in the development of metabolic syndrome indicated by the presence of abdominal obesity, dyslipidaemia, glucose intolerance and increased blood pressure. Epicatechin treatment was found to enhance the oxidative stress status in HFHC groups through an increase in serum nitric oxide levels and a decrease in 8-isoprostane concentrations. Furthermore, WKY-HFHC rats displayed a decrease in IL-6 levels. The lipid profiles in HFHC groups showed improvement, with a decrease in LDL-cholesterol and TAG and an increase in HDL-cholesterol levels observed in WKY-HFHC rats. However, epicatechin was not effective in preventing weight gain, glucose intolerance or hypertension in HFHC fed rats. Overall, the results of this study suggest that epicatechin has the potential to improve the underlying mechanisms associated with metabolic syndrome in obese rats

Parent\u27s views of the importance of making changes in settings where children spend time to prevent obesity

Aim : To examine the kinds of changes parents would like to see in those settings where children spend time (kindergartens and schools, child care centres and after-school care facilities, and the local neighbourhood) in policies and practices that impact on children&rsquo;s risk of obesity, and to establish whether parents might be willing to advocate for changes in these settings. Materials and Methods : 175 parents from five randomly selected primary schools and five randomly selected kindergartens located in suburbs of metropolitan Melbourne completed a questionnaire in which they rated the importance of a number of potential changes to promote healthy eating and increase physical activity in their children. Results : Parents of children in kindergarten most commonly rated changes to the eating environment as important. In contrast, parents of primary school children believed changes related to both eating and physical activity in school were important. Ninety-five per cent of parents of kindergarten children and 89% of parents of primary school children believed it was possible for parents to bring about change to provide more opportunities for their child to eat more healthily and be more physically active. One in four parents reported that they had thought about or had tried to bring about changes in their community. Conclusions : The findings suggest that mobilising parents to take an active role in advocating for change in those settings that have the potential to shape their children&rsquo;s physical activity and eating behaviours may be feasible.<br /

Tallgrass Prairie Center: Effects of Planting Time and Grass-Forb Seeding Ration on Establishment in CRP Pollinator Habitat

Results of an investigation on the creation of habitat for insect pollinators in eastern Iowa

SATLASS (Satellite Autonomous Launch and Assembly)

CubeSats are small satellites composed of U’s. Each U of a CubeSat is approximately 100 x 100 x 113.5 mm and should weigh no more than 2 kg. CubeSats are predominantly used in Low Earth Orbit (LEO), but they have been expanded to use in interplanetary missions. Since the design, manufacturing, and launch of CubeSats are much cheaper than larger satellites (around $80,000 dollars for a 1U CubeSat as opposed to millions of dollars for a traditional satellite), a CubeSat is an attainable goal for a student-led team. CubeSats in the early 2000’s were university or research applications, but starting in the 2010’s they were expanded into the commercial sector. SATLASS aims to act as a deployment method for CubeSats. SATLASS intends to deploy up to three individual satellites to three individual orbits within a 70 kilometer range from the ISS (408.773 km). SATLASS uses cold gas thrusters to maneuver between deployment orbits. While most other deployers operate from ground to orbit, SATLASS starts and ends its mission at a parking station, where it refuels for the next mission so that it can be reused

Medication Gaps and Antipsychotic Polypharmacy in Previously Hospitalized Schizophrenia Patients: An Electronic Cohort Study in Three Canadian Provinces

Background: Real world evidence about antipsychotics focuses on rehospitalization. Modeling the time course of pharmacotherapy would show patients\u27 adherence to medications and physicians\u27 adherence to medication guidelines. We aimed to calculate the cumulative time spent in second generation antipsychotics (SGAs), gaps, antipsychotic polypharmacy, and clozapine in discharged schizophrenia patients. Methods: Hospitalization and pharmacy dispensing data from 2008–2018 in Manitoba, Saskatchewan, and British Columbia were linked and an electronic cohort (N = 2,997) was created (mean follow-up: 49 months, SD = 38). Cohort members were required to have a minimum of 6 weeks medicated with aripiprazole, olanzapine, paliperidone, quetiapine, risperidone, or ziprasidone. Results: The multistate model predicted that schizophrenia patients accumulated 44 months in SGA monotherapy, 4 months in polypharmacy, 11 months in medication gaps and 17 days in clozapine over a 5-year period. The majority of transitions were between SGA and medication gap. Accumulated time in medication gaps was seven times as much as in clozapine. Each 10% delay in SGA initiation post-discharge was associated with a 2, 1, and 6% higher risk for polypharmacy (95% CI: 1.01–1.02), gap (95% CI: 1.01–1.01), and clozapine (95% CI: 1.04–1.08), respectively. Interpretation: Schizophrenia patients accumulated more time unmedicated and in polypharmacy compared to clozapine. Either treatment guidelines for schizophrenia are not followed, or real-world challenges hamper their implementation

Can an incentive-based intervention increase physical activity and reduce sitting among adults? the ACHIEVE (Active Choices IncEntiVE) feasibility study

BackgroundDespite recent interest in the potential of incentivisation as a strategy for motivating healthier behaviors, little remains known about the effectiveness of incentives in promoting physical activity and reducing sedentary behavior, and improving associated health outcomes.This pre-post-test design study investigated the feasibility, appeal and effects of providing non-financial incentives for promoting increased physical activity, reduced sedentary time, and reduced body mass index (BMI) and blood pressure among inactive middle-aged adults.MethodsInactive men (n&thinsp;=&thinsp;36) and women (n&thinsp;=&thinsp;46) aged 40&ndash;65 years were recruited via a not-for-profit insurance fund and participated in a 4 month pre-post design intervention. Baseline and post-intervention data were collected on self-reported physical activity and sitting time (IPAQ-Long), BMI and blood pressure. Participants were encouraged to increase physical activity to 150 mins/week and reduce sedentary behavior by 150 mins/week in progressive increments. Incentives included clothing, recipe books, store gift vouchers, and a chance to win one of four Apple iPad Mini devices. The incentive component of the intervention was supported by an initial motivational interview and text messaging to encourage participants and provide strategies to increase physical activity and reduce sedentary behaviors.ResultsOnly two participants withdrew during the program, demonstrating the feasibility of recruiting and retaining inactive middle-aged participants. While two-thirds of the sample qualified for the easiest physical activity incentive (by demonstrating 100 mins physical activity/week or 100 mins reduced sitting time/week), only one third qualified for the most challenging incentive. Goals to reduce sitting appeared more challenging, with 43% of participants qualifying for the first incentive, but only 20% for the last incentive. More men than women qualified for most incentives. Mean leisure-time physical activity increased by 252 mins/week (leisure-time), with 65% of the sample achieving at least 150 mins/week; and sitting time decreased by 3.1 h/day (both p&thinsp;&lt;&thinsp;0.001) between baseline and follow-up. BMI, systolic and diastolic (men only) blood pressure all significantly decreased. Most participants (50&ndash;85%) reported finding the incentives and other program components helpful/motivating.ConclusionsAcknowledging the uncontrolled design, the large pre-post changes in behavioral and health-related outcomes suggest that the ACHIEVE incentives-based behavior change program represents a promising approach for promoting physical activity and reducing sitting, and should be tested in a randomized controlled trial.<br /

Rheumatic heart disease in pregnancy: strategies and lessons learnt implementing a population-based study in Australia

Background The global burden of rheumatic heart disease (RHD) is two-to-four times higher in women, with a heightened risk in pregnancy. In Australia, RHD is found predominantly among Aboriginal and Torres Strait Islander peoples. Methods This paper reviews processes developed to identify pregnant Australian women with RHD during a 2-year population-based study using the Australasian Maternity Outcomes Surveillance System (AMOSS). It evaluates strategies developed to enhance reporting and discusses implications for patient care and public health. Results AMOSS maternity coordinators across 262 Australian sites reported cases. An extended network across cardiac, Aboriginal and primary healthcare strengthened surveillance and awareness. The network notified 495 potential cases, of which 192 were confirmed. Seventy-eight per cent were Aboriginal and/or Torres Strait Islander women, with a prevalence of 22 per 1000 in the Northern Territory. Discussion Effective surveillance was challenged by a lack of diagnostic certainty, incompatible health information systems and varying clinical awareness among health professionals. Optimal outcomes for pregnant women with RHD demand timely diagnosis and access to collaborative care. Conclusion The strategies employed by this study highlight gaps in reporting processes and the opportunity pregnancy provides for diagnosis and re/engagement with health services to support better continuity of care and promote improved outcomes.The authors gratefully acknowledge aid from the Australian National Health and Medical Research Council (NHMRC) project grant #1024206 and NHMRC Postgraduate Scholarship #11332944; University of Technology Sydney Chancellor’s Research Scholarship; and END RHD Centre of Research Excellence, Telethon Kids Institute, University of Western Australia

The Functional Basis for Hemophagocytic Lymphohistiocytosis in a Patient with Co-inherited Missense Mutations in the Perforin (PFN1) Gene

About 30% of cases of the autosomal recessive immunodeficiency disorder hemophagocytic lymphohistiocytosis are believed to be caused by inactivating mutations of the perforin gene. We expressed perforin in rat basophil leukemia cells to define the basis of perforin dysfunction associated with two mutations, R225W and G429E, inherited by a compound heterozygote patient. Whereas RBL cells expressing wild-type perforin (67 kD) efficiently killed Jurkat target cells to which they were conjugated, the substitution to tryptophan at position 225 resulted in expression of a truncated (∼45 kD) form of the protein, complete loss of cytotoxicity, and failure to traffic to rat basophil leukemia secretory granules. By contrast, G429E perforin was correctly processed, stored, and released, but the rat basophil leukemia cells possessed reduced cytotoxicity. The defective function of G429E perforin mapped downstream of exocytosis and was due to its reduced ability to bind lipid membranes in a calcium-dependent manner. This study elucidates the cellular basis for perforin dysfunctions in hemophagocytic lymphohistiocytosis and provides the means for studying structure–function relationships for lymphocyte perforin