A fast protocol for purification of translating mRNAs

We present a protocol that enables easy extraction of translating mRNAs by purification of tagged ribosomes and associated mRNAs. By studying three different examples of stress influenced genes, we demonstrate the effectiveness and compare our rapid method to an older protocol (Halbeisen et al., 2009).

Cognitive-Behavioural Analysis System of Psychotherapy (CBASP), a drug, or their combination: differential therapeutics for persistent depressive disorder: a study protocol of an individual participant data network meta-analysis.

INTRODUCTION Despite important advances in psychological and pharmacological treatments of persistent depressive disorders in the past decades, their responses remain typically slow and poor, and differential responses among different modalities of treatments or their combinations are not well understood. Cognitive-Behavioural Analysis System of Psychotherapy (CBASP) is the only psychotherapy that has been specifically designed for chronic depression and has been examined in an increasing number of trials against medications, alone or in combination. When several treatment alternatives are available for a certain condition, network meta-analysis (NMA) provides a powerful tool to examine their relative efficacy by combining all direct and indirect comparisons. Individual participant data (IPD) meta-analysis enables exploration of impacts of individual characteristics that lead to a differentiated approach matching treatments to specific subgroups of patients. METHODS AND ANALYSIS We will search for all randomised controlled trials that compared CBASP, pharmacotherapy or their combination, in the treatment of patients with persistent depressive disorder, in Cochrane CENTRAL, PUBMED, SCOPUS and PsycINFO, supplemented by personal contacts. Individual participant data will be sought from the principal investigators of all the identified trials. Our primary outcomes are depression severity as measured on a continuous observer-rated scale for depression, and dropouts for any reason as a proxy measure of overall treatment acceptability. We will conduct a one-step IPD-NMA to compare CBASP, medications and their combinations, and also carry out a meta-regression to identify their prognostic factors and effect moderators. The model will be fitted in OpenBUGS, using vague priors for all location parameters. For the heterogeneity we will use a half-normal prior on the SD. ETHICS AND DISSEMINATION This study requires no ethical approval. We will publish the findings in a peer-reviewed journal. The study results will contribute to more finely differentiated therapeutics for patients suffering from this chronically disabling disorder. TRIAL REGISTRATION NUMBER CRD42016035886.

Effects of Estrogen on Muscle Damage in Response to an Acute Resistance Exercise Protocol

Creatine Kinase (CK) is used as a measure of exercise-induced muscle membrane damage. During acute eccentric (muscle lengthening) exercise, muscle sarcolemma, sarcoplasmic reticulum, and Z-lines are damaged, thus causing muscle proteins and enzymes to leak into the interstitial fluid. Strenuous eccentric exercise produces an elevation of oxygen free radicals, which further increases muscle damage. Muscle soreness and fatigue can be attributed to this membrane damage. Estradiol, however, may preserve membrane stability post-exercise (Brancaccio, Maffulli, & Limongelli, 2007; Carter, Dobridge, & Hackney, 2001; Tiidus, 2001). Because estradiol has a similar structure to Vitamin E, which is known to have antioxidant properties, and both are known to affect membrane structure, researchers have proposed that estrogen acts as an antioxidant to provide a protective effect on the post-exercise muscle of women (Sandoval & Matt, 2002). As a result, it has been postulated that muscles in women incur less damage in response to an acute strenuous exercise as compared to men. PURPOSE: To determine if circulating estrogen concentrations are related to muscle damage, as measured by creatine kinase activity and to determine gender differences in creatine kinase as a marker of muscle damage in response to an acute heavy resistance exercise protocol. METHODS: 7 healthy, resistance-trained, eumenhorrheic women (23±3 y, 169±9.1 cm, 66.4±10.5 kg) and 8 healthy, resistance-trained men (25±5 y, 178±6.7 cm, 82.3±9.33 kg) volunteered to participate in the study. Subjects performed an Acute Resistance Exercise Test (ARET) consisting of 6 sets of 5 repetitions Smith machine squats at 90% of their previously determined 1-RM. Blood samples were taken pre-, mid-, post-, 1 hour post-, 6 hours post-, and 24 hours post-exercise. Samples were stored at -80ºC until analyzed. Serum creatine kinase was measured using an assay kit from Genzyme (Framingham, MA). Serum estradiol was measured by an ELISA from GenWay (San Diego, CA). Estradiol b-receptor presence on granulocytes was measured via flow cytometry using primary antibodies from Abcam (Cambridge, MA) and PeCy7 antibodies (secondary) from Santa Cruz (Santa Cruz, CA). RESULTS: No significant correlations between estrogen and CK response were found after an acute resistant exercise protocol. Moreover, no significant change in estradiol receptors were expressed on granulocytes after exercise. Creatine Kinase response, however, differed significantly between genders. Men had higher resting CK concentrations throughout all time points. Creatine Kinase response increased significantly after exercise in both men and women (p=0.008, F=9.798). Men had a significantly higher CK response at 24 hours post exercise than women. A significant condition/sex/time interaction was exhibited in CK response (p=0.02, F=4.547). Perceived general soreness presented a significant condition, sex interaction (p=0.01, F=9.532). DISCUSSION: Although no estradiol and CK response correlations were found in response to exercise, a significant difference in creatine kinase activity was present between men and women. This discrepancy of our results and findings in the literature may be due to the high variability between subjects in creatine kinase activity as well as estrogen concentrations. The lack of significance in change of estradiol receptor expression on granulocytes in response to exercise may be due to intracellular estradiol receptor staining and non-specific gating for granulocytes rather than additional staining for neutrophil markers. Because neutrophils are the initial cells present in the inflammatory response after strenuous exercise, staining for estrogen receptors on this cell type may allow for a better understanding of the effect of estrogen and its hypothesized protective effect against muscle damage. Furthermore, the mechanism of action may include estradiol receptor expression on the muscle fiber itself may play a role in the protective effects of estradiol rather than or in addition to expression on neutrophils. We have shown here that gender differences occur in CK activity as a marker of muscle damage in response to strenuous eccentric exercise, but may not be the result of estradiol concentration or estradiol receptor expression on granulocytes. Other variables should be examined in order to determine the mechanism involved in the difference in creatine kinase as a marker of muscle damage between men and women after heavy resistance exercise.

Physician's adherence to the standard protocol for diabetes treatment in Brooke Army Medical Center (BAMC)

The level of compliance with clinical practice guidelines for patients with Type II Diabetes Mellitus was evaluated in 157 patients treated at BAMC from 1 January 2006 to 1 January 2007. This retrospective analysis was conducted reviewing data from medical records and following the VA/DOD protocols that health care providers are expected to follow at this facility. Data collected included patient’s age and gender, presence or absence of complications of diabetes, physical examination findings, glycemic and lipid control, eye care, foot care, kidney function, and self-management and education. Subjects were selected performing systematic random sampling, and included both male and female patients, from a variety of ages and ethnic groups. The Diabetes complications screened for included glycemic and lipid complications, retinopathy, cardiovascular complications, peripheral circulation complications, and nephropathy. The results revealed that 19.10% had no complications and that the most common complications were: cardiovascular (49.68%), glycemic and lipid control (10.82%), retinopathy and peripheral circulation (8.28% each), and nephropathy (2.54%). Only 2.54% of the records reviewed did not include information on complications. Strictly following the Department of Defense guidelines, six treatment modules were evaluated independently and together to get a final percentage of adherence to the clinical practice guidelines. It was established that the level of adherence was going to be graded as follows: Extremely deficient: 0-15%; very poor: 16-30%; Poor and in need of improvement: 31-45%. Acceptable: 46-60%; Good: 61-80%, and Excellent: 81-100%. The results indicated that the percentage of physicians' adherence to each protocol was as follows: 88.31%, 89.93%, 90.63%, 89.42%, 89.42% and 89.64%. When the results were pooled, the level of adherence to the clinical practice guidelines was 89.55%, proving my hypothesis that Brooke Army Medical Center physicians have excellent adherence to the standard protocols for Diabetes Type II to treat their patients. ^

A simplified protocol for the treatment of acute radiation overexposures from sources external to the body

When compared to other types of occupational injuries, radiation overexposure events are somewhat rare, so health care providers may not be familiar with the actual clinical care to be provided when such an event occurs. Radiation overexposure treatment decisions are predicated on the amount of radiation dose received, which is a value many health care providers may not have the knowledge or expertise to either calculate or even estimate. Even the different units of measure for radiation exposure and dose received can be a source of confusion. The prompt treatment of radiation overexposure victims could be enhanced and facilitated through the creation of a single, simple protocol that consists of the various means of dose measurement and estimation, correlated to the corresponding appropriate clinical care measures. This culminating experience will assemble essential information currently maintained in disparate references to create a single, simplified protocol to facilitate the treatment of victims of acute external radiation overexposure. ^