Clamp loading,unloading and intrinsic stability of the PCNA, β and gp45 sliding clamps of human,E. coli and T4 replicases

Background: The high speed and processivity of replicative DNA polymerases reside in a processivity factor which has been shown to be a ring-shaped protein. This protein (‘sliding clamp’) encircles DNA and tethers the catalytic unit to the template. Although in eukaryotic, prokaryotic and bacteriophage-T4 systems, the processivity factors are ring-shaped, they assume different oligomeric states. The Escherichia coli clamp (the β subunit) is active as a dimer while the eukaryotic and T4 phage clamps (PCNA and gp45, respectively) are active as trimers. The clamp can not assemble itself on DNA. Instead, a protein complex known as a clamp loader utilizes ATP to assemble the ring around the primer-template. This study compares properties of the human PCNA clamp with those of E. coli and T4 phage. Results: The PCNA ring is a stable trimer down to a concentration below 100 nm (K_d ≈ 21 nm ). On DNA, the PCNA clamp slides freely and dissociates from DNA slowly (t_1/2 ≈ 24 min). β is more stable in solution (K_d < 60 pm ) and on DNA (t_1/2 ≈ 1 h) than PCNA which may be explained by its simpler oligomeric state. The T4 gp45 clamp is a much less stable trimer than PCNA (K_d ≈ 250 nm ) and requires association with the polymerase to stabilize it on DNA as observed previously. The consequence of this cooperation between clamp and polymerase is that upon finishing a template and dissociation of the polymerase from DNA, the gp45 clamp spontaneously dissociates from DNA without assistance. However, the greater stability of the PCNA and β clamps on DNA necessitates an active process for their removal. The clamp loaders (RF-C and γ complex) were also capable of unloading their respective clamps from DNA in the presence of ATP. Conclusions: The stability of the different clamps in solution correlates with their stability on DNA. Thus, the low stability of the T4 clamp explains the inability to isolate gp45 on DNA. The stability of the PCNA and β clamps predicts they will require an unloading factor to recycle them on and off DNA during replication. The clamp loaders of PCNA and β double as clamp unloaders presumably for the purpose of clamp recycling.     

The study of tuberculosis outbreak in a high school—Shanghai,China, 2017–2018

Journal of Public Health - To investigate the attack rate of active tuberculosis (TB) cases and detection rate of latent tuberculosis infection (LTBI) cases, and to identify possible factors...     

Machine Learning Algorithms to Predict Mortality and Allocate Palliative Care for Older Patients With Hip Fracture

ObjectivesTo evaluate a machine learning model designed to predict mortality for Medicare beneficiaries aged >65 years treated for hip fracture in Inpatient Rehabilitation Facilities (IRFs).DesignRetrospective design/cohort analysis of Centers for Medicare & Medicaid Services Inpatient Rehabilitation Facility–Patient Assessment Instrument data.Setting and ParticipantsA total of 17,140 persons admitted to Medicare-certified IRFs in 2015 following hospitalization for hip fracture.MeasuresPatient characteristics include sociodemographic (age, gender, race, and social support) and clinical factors (functional status at admission, chronic conditions) and IRF length of stay. Outcomes were 30-day and 1-year all-cause mortality. We trained and evaluated 2 classification models, logistic regression and a multilayer perceptron (MLP), to predict the probability of 30-day and 1-year mortality and evaluated the calibration, discrimination, and precision of the models.ResultsFor 30-day mortality, MLP performed well [acc = 0.74, area under the receiver operating characteristic curve (AUROC) = 0.76, avg prec = 0.10, slope = 1.14] as did logistic regression (acc = 0.78, AUROC = 0.76, avg prec = 0.09, slope = 1.20). For 1-year mortality, the performances were similar for both MLP (acc = 0.68, AUROC = 0.75, avg prec = 0.32, slope = 0.96) and logistic regression (acc = 0.68, AUROC = 0.75, avg prec = 0.32, slope = 0.95).Conclusion and ImplicationsA scoring system based on logistic regression may be more feasible to run in current electronic medical records. But MLP models may reduce cognitive burden and increase ability to calibrate to local data, yielding clinical specificity in mortality prediction so that palliative care resources may be allocated more effectively.     

大鼠不同程度的急性颅脑损伤后血糖浓度的变化及其预后

目的 :探讨大鼠不同程度的急性颅脑损伤后血糖浓度的变化及其与预后的关系。方法 :Wistar雄性大鼠 2 0只 ,随机分为 2组 ,A组 :将 5 0 0 g砝码由 35 cm高度自由下落打击大鼠头部 ;B组 :将 5 0 0 g砝码由 2 0 cm高度自由下落打击大鼠头部 ;设自身对照 ,分别测定不同伤情的两组动物伤前及伤后血糖浓度的变化及观察 2 4h内两组动物死亡情况。结果 :A、B组伤后测得的血糖值均显著高于正常值 (P <0 .0 5 ) ;且 A组伤后测得的血糖值又显著高于 B组 (P <0 .0 5 ) ;A组动物死亡 8只 ,B组死亡 1只。结论 :急性颅脑损伤后均有不同程度的血糖升高 ,而且颅脑损伤程度越重 ,血糖值越高 ,死亡率越高。     

醒脑静在重症一氧化碳中毒抢救中的临床应用

目的　探讨醒脑静对急性重症一氧化碳 (CO)中毒的催醒作用。方法　在急性重症CO中毒常规综合抢救中加入醒脑静并与常规方法进行对比研究。结果　醒脑静治疗组较常规治疗组清醒时间明显缩短 (P <0 .0 1)。结论　醒脑静可作为急性重症CO中毒病人综合抢救中常规用药。     

乳腺髓样癌的超声检查与组织病理的相关性分析

目的:探讨超声检查在诊断乳腺典型髓样癌中的临床意义。方法:按严格的病理诊断学标准将病例重新分类为乳腺典型髓样癌和非典型髓样癌,比较两者间声像学特征的异同。结果:32例中有13侧重新诊断为乳腺典型髓样癌;另外19例为非典型髓样癌。13例典型髓样癌患者中的10例边界整齐光滑;全部19例非典型髓样癌病人没有一例边界光滑,或者边界呈锯齿状(16例),或者边界光滑伴局部不规则(3例)。非典型与典型髓样癌间肿瘤边界的规则性有明显的差异。结论:如果超声检查提示肿瘤边界呈锯齿状或肿瘤边界光滑伴局部不规则,应行整个肿瘤的病理评估防止典型髓样癌的过诊断。     

电磁辐射对小鼠神经系统超微结构影向的分析

目的探讨移动电话的电磁辐射对生物体的影响.方法用大众常用移动电话作为辐射源,工作频率为900MHz,功率密度为1190μW/cm2,在一定范围对小鼠辐射2h/d,30d后把小鼠断颈处死,取出大脑皮层、海马和小脑进行电镜观察分析.结果电镜所见,处理组与对照组小鼠的神经系统的细胞超微结构未见明显异常.结论一定时间内,移动电话的电磁辐射,对小鼠神经系统的细胞超微结构并没有明显影响.     

Dark Tea: A popular beverage with possible medicinal application

Tea is a famous beverage that is produced from leaves of Camellia sinensis. Amongst the six major tea categories in China, dark tea is the only one that involves microbial fermentation in the manufacturing process, which contributes unique flavors and functions for the tea. In the recent decade, the reports about the biofunctions of dark teas have increased rapidly. Therefore it may be the proper time to consider dark tea as one potential homology of medicine and food. In this viewpoint, our current understanding of the chemical constituents, biological activities and possible health beneficial effects of dark teas were introduced. Some future directions and challenges to the development perspectives of dark teas were also discussed.