Reactivation of heat-inactivated Ku proteins by heat shock cognate protein HSC73

Purpose: Mouse double-stranded DNA-dependent protein kinase (DNA-PK) activity is heat sensitive. Recovery of heat-inactivated DNA repair activity is a problem after combination therapy with radiation and heat. We investigated the mechanism of recovery of heat-inactivated DNA-PK activity.

Methods: Hybrid cells containing a fragment of human chromosome 8 in scid cells (RD13B2) were used. DNA-PK activity was measured by an in vitro assay. Immunoprecipitation of the nuclear extract was performed with an anti-Ku80 antibody. Proteins co-precipitated with Ku80 were separated by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and detected by Western blotting using anti-heat shock protein (HSP)72 and anti-heat shock cognate protein (HSC)73 antibodies. HSC73 was overexpressed with the pcDNA3.1 vector. Short hairpin (sh)RNA was used to downregulate HSC73 and HSP72.

Results: The activity of heat-inactivated DNA-PK recovered to about 50% of control during an additional incubation at 37?°C after heat treatment at 44?°C for 15?min in the presence of cycloheximide (which inhibits de novo protein synthesis). Maximal recovery was observed within 3?h of incubation at 37?°C after heat treatment. Constitutively expressed HSC73, which folds newly synthesized proteins, reached maximal levels 3?h after heat treatment using a co-immunoprecipitation assay with the Ku80 protein. Inhibiting HSC73, but not HSP72, expression with shRNA decreased the recovery of DNA-PK activity after heat treatment.

Conclusions: These results suggest that de novo protein synthesis is unnecessary for recovery of some heat-inactivated DNA-PK. Rather, it might be reactivated by the molecular chaperone activity of HSC73, but not HSP72.     

Are primary care physicians ill equipped to evaluate and treat childhood physical inactivity?

ABSTRACT

Objective

To investigate primary care physician clinical practice patterns, barriers, and education surrounding pediatric physical activity (PA), and to compare practice patterns by discipline.     

Intra-articular Hymovis injection for managing hip OA in active sportsmen. A 24-month observational retrospective clinical investigation

BackgroundHip Osteoarthitis (OA) risk is sport-specific and depends on frequency, intensity, and type of mechanic stress the hip is subjected to. This retrospective observational study aims to investigate the safety and performance of Hymovis (HYADD-4) injection, a hexadecyl (C-16) HA-derivative, when used to manage symptomatic hip OA in active middle-aged sportsmen over a 24-month observation period.MethodsThe retrospective analysis included clinical records of active sportsmen, aged between 40 and 65 years, and suffering from symptomatic Kellgren-Lawrence grade II to III hip OA, treated with two (24 mg/3 ml) Hymovis injections, two weeks apart, every 3–4 months, for two years. When available, data on MRI examination were included in the analysis as well as Heidelberg Sports Activity Score (HAS) and Copenhagen Hip and Groin Outcome Score (HAGOS) questionnaires.ResultsThirty patients (56.4 ± 7.3 years) were included in the study, sixteen cyclists and 14 tennis players. For all patients, HAS and most HAGOS scores improved significantly (p < 0.05) at the first control visit (4 months) and further improved over time. For all other scores an important clinical benefit was experienced by more than 50% of participants. No adverse events were recorded.ConclusionTreatment of hip OA in active sportsmen with Hymovis seems a safe and effective approach for the management of OA symptoms, by potentially protecting cartilage and subchondral bone from further damage.     

Effects of exogenous factors on spatial accuracy in neurosurgery

The study aimed to assess the effect of exogenous factors such as surgeon posture, surgical instrument length, fatigue after a night shift, exercise and caffeine consumption on the spatial accuracy of neurosurgical manipulations. For the evaluation and simulation of neurosurgical manipulations, a testing device developed by the authors was used. The experimental results were compared using nonparametric analysis (Wilcoxon test) and multivariate analysis, which was performed using mixed models. The results were considered statistically significant at p < 0.05. The study included 11 first-year neurosurgery residents who met the inclusion criteria. Hand support in the sitting position (Wilcoxon test p value = 0.0033), caffeine consumption (p = 0.0058) and the length of the microsurgical instrument (p = 0.0032) had statistically significant influences on the spatial accuracy of surgical manipulations (univariate analysis). The spatial accuracy did not significantly depend on the type of standing position (Wilcoxon test p value = 0.2860), whether the surgeon was standing/sitting (p = 0.1029), fatigue following a night shift (p = 0.3281), or physical exertion prior to surgery (p = 0.2845).When conducting the multivariate analysis, the spatial accuracy significantly depended on the test subject (p < 0.0001), the use of support during the test (p = 0.0001), and the length of the microsurgical instrument (p = 0.0397). To increase the spatial accuracy of microsurgical manipulations, hand support and shorter tools should be used. Caffeine consumption in high doses should also be avoided prior to surgery.     

Measuring disease activity and response to treatment in rheumatoid arthritis

Introduction: Effective treatment of rheumatoid arthritis (RA) requires suppression of the underlying inflammation. Measurement of such inflammation, the disease activity, is mandatory to target treatment and maximize outcomes. However, this is not as straightforward as it may seem.

Areas covered: The many tools developed to measure disease activity in RA, from composite scores and patient-reported outcomes, to laboratory markers and imaging are discussed, with a focus on their utility in guiding therapy and assessing response. The complex issues in measuring disease activity in RA, whether in clinical trials or normal clinical practice, and in the context of national guidelines and recommendations, available time, and resources are considered.

Expert commentary: The key to effective management of RA is the rapid suppression of inflammation, ideally to remission, with maintenance of such remission. The aim is to prevent disability and maximize quality of life. Central to this is the ability to determine disease activity (potentially open to suppression) as opposed to damage (irreversible). A variety of measures are currently available, allowing better assessment of response to treatment. In the future, the development of predictive biomarkers allowing targeting of drugs may revolutionize this field and render the tools of today redundant.     

Making meaning of the virginity experience: young men's perceptions in the United States

ABSTRACT

A qualitative grounded theory approach was used to analyze 34 narratives from young men about their experience of retaining, losing, or giving away their virginity, where 7 said they were virgins, 26 said they were non-virgins, and one did not mention his virginity status. We found that virginity is a multidimensional concept, with two distinct dimensions: the experiential and the developmental. The experiential dimension refers to young men's perception and understanding of their virginity in four overlapping areas: physical, spiritual, relational, and emotional. The developmental dimension refers to young men's perception of their underlying sexual identity growth processes and the gendered and sexual double standards that influence this understanding. Clinical implications of this study are useful in education and counseling. They seek to empower young men with information relating to the multidimensional nature of virginity experience, and minimize the negative effects on romantic relationships, sexual adjustment, and self-identity development. The findings are helpful in designing interventions for young men and women who are at high risk for sexually transmitted infections and can help them negotiate mutually safe and pleasurable experiences regarding their virginity.     

基于网络药理学的气滞胃痛颗粒抗炎镇痛活性作用分析

目的通过网络药理学的方法预测气滞胃痛颗粒抗炎镇痛主要活性成分的作用靶点,结合中医方解配伍理论对其多成分-多靶点-多通路的作用进行分析。方法基于TCMSP中药系统生物学分析数据库收集气滞胃痛颗粒中6味中药的主要化学成分,并借助LC-MS技术对所筛成分进行分析,通过TCMSP检索和Pharmmapper软件预测获取各成分主要的作用靶标,并通过DIP数据库,利用蛋白质相互作用信息建立药物靶标与炎症疼痛靶标的关联,构建药物-靶标-疾病网络,通过网络特征分析气滞胃痛颗粒抗炎镇痛的作用靶标,阐释其抗炎镇痛的主要作用机制。结果根据网络分析,共有44个炎症疼痛靶点与气滞胃痛颗粒密切相关,其中直接作用靶点有20个,主要是对环加氧酶-2(COX-2)和诱导型一氧化氮合酶(i NOS)等蛋白酶的作用,作用机制可能与调节肿瘤坏死因子(TNF)信号通路、NOD样受体(NLR)信号通路、血管内皮生长因子(VEGF)信号通路等与炎症疼痛密切相关的信号通路有关。结论气滞胃痛颗粒抗炎镇痛作用体现了中药多成分、多靶点、多途径的作用特点,该研究为深入阐释气滞胃痛颗粒抗炎镇痛作用机制提供科学依据,并且进一步说明了中医药古方配伍理论的科学性。