Miracle-dz/UniFAR-FTUs huggingface dataset:
Construction and properties of a recombinant herpes simplex virus 1 lacking both S-component origins of DNA synthesis

The herpes simplex virus 1 (HSV-1) genome contains three origins of DNA synthesis (Ori) utilized by viral DNA synthesis proteins. One sequence (OriI) maps in the L component, whereas two sequences (OriS) map in the S component. We report the construction of a recombinant virus, R7711, from which both OriS sequences have been deleted, and show that the OriS sequences are not essential for the replication of HSV-1 in cultured cells. In addition to the deletions of OriS in R7711, the alpha 47 gene and the 5' untranscribed and transcribed noncoding regions of the U(S)11 gene were deleted, one of the alpha 4 promoter-regulatory regions was replaced with the simian virus 40 promoter, and the alpha 22 promoter was substituted with the alpha 27 promoter. The total amount of viral DNA synthesized in Vero cells infected with the OriS-negative (OriS-) virus was approximately that seen in cells infected with the OriS-positive virus. However, cells infected with the OriS- virus accumulated viral DNA more slowly than those infected with the wild-type virus during the first few hours after the onset of DNA synthesis. In single-step growth experiments, the yield of OriS- progeny virus was reduced at most fourfold. Although a single OriS (R. Longnecker and B. Roizman, J. Virol. 58:583-591, 1986) and the single OriL (M. Polvino-Bodnar, P. K. Orberg, and P. A. Schaffer, J. Virol. 61:3528-3535, 1987) have been shown to be dispensable, this is the first indication that both copies of OriS are dispensable and that one copy of an Ori sequence may suffice for the replication of HSV-1.
Modeling spatial distribution of oxygen in 3d culture of islet beta‐cells

Three‐dimensional (3D) scaffold culture of pancreatic β‐cell has been proven to be able to better mimic physiological conditions in the body. However, one critical issue with culturing pancreatic β‐cells is that β‐cells consume large amounts of oxygen, and hence insufficient oxygen supply in the culture leads to loss of β‐cell mass and functions. This becomes more significant when cells are cultured in a 3D scaffold. In this study, in order to understand the effect of oxygen tension inside a cell‐laden collagen culture on β‐cell proliferation, a culture model with encapsulation of an oxygen‐generator was established. The oxygen‐generator was made by embedding hydrogen peroxide into nontoxic polydimethylsiloxane to avoid the toxicity of a chemical reaction in the β‐cell culture. To examine the effectiveness of the oxygenation enabled 3D culture, the spatial‐temporal distribution of oxygen tension inside a scaffold was evaluated by a mathematical modeling approach. Our simulation results indicated that an oxygenation‐aided 3D culture would augment the oxygen supply required for the β‐cells. Furthermore, we identified that cell seeding density and the capacity of the oxygenator are two critical parameters in the optimization of the culture. Notably, cell‐laden scaffold cultures with an in situ oxygen supply significantly improved the β‐cells' biological function. These β‐cells possess high insulin secretion capacity. The results obtained in this work would provide valuable information for optimizing and encouraging functional β‐cell cultures. © 2016 American Institute of Chemical Engineers Biotechnol. Prog. , 33:221–228, 2017
Differential properties of two charge components in frog skeletal muscle.

Charge movement in frog twitch muscle fibres bathed in a moderately hypertonic solution (containing 350 mM‐sucrose) showed a main component (Q beta) and a secondary hump (Q gamma). Assuming that Q beta decays with a single exponential and Q gamma follows the time course of a symmetrical bell, a mathematical model was developed to separate Q gamma from Q beta. The activation curves of Q beta and Q gamma can be individually fitted by the two‐state model of Schneider & Chandler (1973). The voltage distribution of Q gamma is approximately twice as steep as that of Q beta and the maximum moveable amount of Q beta about four times that of Q gamma. The decay rate constant of Q beta, expressed as a function of potential, follows the familiar U‐shaped curve whereas the reciprocal of the time‐to‐peak of Q gamma rises linearly with increasing potentials. Application of the mathematical dissection technique to Qon values in solutions of varying concentrations of tetracaine yielded the dose dependence of the suppression of Q gamma by tetracaine. Q gamma inactivates more steeply than Q beta as the holding potential is made less negative. Each of the steady‐state inactivation curves of Q beta and Q gamma forms a mirror image with the respective activation curve. It is speculated that Q gamma might be more directly involved than Q beta in triggering calcium release and activating contraction.
Influence of the prion protein gene, Prnp, on scrapie susceptibility in sheep

Natural scrapie in sheep occurs through a complex interplay between host genetic elements and various strains of the infectious scrapie agent. Scrapie‐related polymorphisms in the coding region of the prion protein (PrP) gene, Prnp, have been studied in a number of breeds. The disease‐promoting V136 allele, and the susceptibility‐reducing R171 allele, have proved to be most important. However, variation in the coding region of Prnp cannot alone explain the diverse patterns of scrapie susceptibility in various breeds. For instance, in many breeds plagued with scrapie, the V136 allele appears to be a rarity. The R171 allele greatly reduces scrapie susceptibility. This lays the molecular foundation for marker‐assisted breeding for reduced scrapie susceptibility now underway in many countries. Although potentially important, and still under investigation, variable expression level and pattern of the ovine Prnp appears to be of little importance for the occurrence of natural scrapie. Studies of scrapie in mice also indicate that genetic elements other than Prnp may have a strong influence on scrapie incubation time, and hence susceptibility. Narrowing down the search to focus on these elements and identification of candidate genes are important tasks for future research in sheep scrapie.
The effect of pretreatment with clonidine on propofol consumption in opium abuser and non-abuser patients undergoing elective leg surgery

Objective: Clonidine, an alpha-2 adrenergic agonist, increases the quality of perioperative sedation and analgesia with a few side effects. This study was designed to assess the effect of clonidine premedication on the anesthesics used for elective below knee surgeries in opium abusers and non-abusers. Materials and Methods: In a randomized clinical trial, 160 patients were selected and assigned into four groups. Eighty patients among the opium abusers were divided randomly into clonidine and no clonidine groups, with 40 patients in each, and 80 among the non-abusers were again divided randomly into clonidine and no clonidine groups, with 40 patients in each group. All were anesthetized for elective orthopedic operation using the same predetermined method. The total administered dose of propofol and other variables were compared. Results: The total propofol dose in a decreasing order was as follows: Abuser patients receiving placebo (862 ± 351 mg), non-abuser patients receiving placebo (806 ± 348 mg), abuser patients receiving clonidine (472 ± 175 mg), and non-abuser patients receiving clonidine (448 ± 160 mg). Hence, a statistically significant difference was observed among the four study groups (P value for ANOVA = 0.0001). Conclusion: Adding clonidine as a preoperative medication decreases the patient's anesthetic needs; this decrease was even more considerable on the anesthetic needs than the effect of opium abuse history on anesthetic dose.
Long‐term follow‐up of the Ischemic Heart Disease Life Stress Monitoring Program.

&NA; This study examines patient outcomes up to seven years after a one‐year, post‐myocardial infarction (MI) stress monitoring and management program. A previous randomized controlled trial involving 461 male patients demonstrated that the program lowered stress scores and reduced one‐year cardiac deaths by almost 50%, but made no difference in readmission rates or durations. The present study used medical records to assess the durability of these outcomes. Results indicated that the difference in cardiac deaths was primarily due to out‐of‐hospital deaths (that is, those that were more likely to have been sudden deaths) and persisted for six months beyond the end of the program, after which the mortality curves began to approach one another. In addition, although during the program year MI recurrences occurred at the same rate in the two groups, during the years after the program there were fewer MI recurrences among the treated patients. Over the full follow‐up period the group difference in MI recurrences was statistically significant (p = 0.043). Results are considered in terms of methodological limitations of the study, including the timing of randomization, sample selection requirements, and group differences on baseline variables. Findings are also discussed in relation to the outcomes of other programs producing long‐term reductions in recurrences or deaths from ischemic heart disease. It is suggested that, regardless of their approach, all such programs may share a common therapeutic mechanism: the provision of emotional support during a period of high vulnerability to stress.
Incremental Value of Biochemical and Echocardiographic Measures in Prediction of Ischemic Stroke: The Strong Heart Study

Background and Purpose— American Indians have high rates of stroke. Improved risk stratification could enhance prevention, but the ability of biochemical and echocardiographic markers of preclinical disease to improve stroke prediction is not well-defined. Methods— We evaluated such markers as predictors of ischemic stroke in a community-based cohort of American Indians without prevalent cardiovascular or renal disease. Laboratory markers included C-reactive protein, fibrinogen, urine albumin-to-creatinine ratio, and glycohemoglobin (HbA1c), whereas echocardiographic parameters comprised left atrial diameter, left ventricular mass, mitral annular calcification, and the ratio of early to late mitral diastolic velocities. Predictive performance was judged by indices of discrimination, reclassification, and calibration. Results— After adjustment for standard risk factors, only HbA1c, albuminuria, and left atrial diameter were significantly associated with first ischemic stroke. Addition of HbA1c, although not urine albumin-to-creatinine ratio, to a basic clinical model significantly improved the C-statistic (0.714 versus 0.695; P=0.044), whereas left atrial diameter modestly enhanced integrated discrimination improvement (0.90%; P=0.004), but not the C-statistic (0.701; P=0.528). When combined with HbA1c, left atrial diameter further increased integrated discrimination improvement (1.81%; P<0.001) but not the C-statistic (0.716). No marker achieved significant net reclassification improvement. Conclusions— In this cohort at high cardiometabolic risk, HbA1c emerged as the foremost predictor of ischemic stroke when added to traditional risk factors, affording substantially improved discrimination, with a more modest contribution for left atrial diameter. These findings bolster the role of HbA1c in cardiovascular risk assessment among persons with glycometabolic disorders and provide impetus for further study of the incremental value of echocardiography in high-risk populations.
Assessment of Glucose Turnover Rates in Euglycaemic Clamp Studies using Primed‐constant [3‐3H]‐glucose Infusion and Labelled or Unlabelled Glucose Infusates

Underestimation of glucose turnover rates has been a problem in clamp studies using primed‐constant [3‐3H]‐glucose infusion technique. Due to slow mixing in interstitial compartments concealed specific activity gradients may arise between plasma and interstitial compartments during intravenous unlabelled glucose infusion. Such specific activity gradients, however, can be prevented if plasma specific activity is maintained constant. Two euglycaemic clamp studies (insulin infusion 40 mU m−2 min−1) were performed in six lean normal subjects. Using conventional unlabelled glucose infusates plasma specific activity declined by 74%, tracer determined glucose appearance was smaller than actual glucose infusion rates (317 ± 11 vs 366 ± 15 mg m−2 min−1, p< 0.001), and erroneous negative values were calculated for glucose production (‐49 ± 7 mg m−2 min−1). Average underestimation during the first 2 h correlated with glucose infusion rates (r = 0.88, p < 0.02). In contrast, when plasma specific activity was maintained constant, using appropriately labelled glucose infusates, tracer determined glucose appearance and glucose infusion rates were similar (385 ± 16 vs 385 ± 17 mg m−2 min−1), and negative errors for glucose production were avoided. In conclusion, using unlabelled glucose infusates, as in previous studies, suppression of glucose production is overestimated and stimulation of glucose utilization is underestimated. As errors were greater with larger glucose infusions, the mistakes may have been greatest in insulin sensitive control subjects, and smaller in insulin resistant subjects. Therefore, re‐evaluation of hepatic insulin sensitivity seems appropriate in diabetes, obesity, and other insulin resistant states.
Nuclear Export of the Oncoprotein v-ErbA Is Mediated by Acquisition of a Viral Nuclear Export Sequence*

v-ErbA, an oncogenic derivative of the thyroid hormone receptor α (TRα) carried by the avian erythroblastosis virus, contains several alterations including fusion of a portion of avian erythroblastosis virus Gag to its N terminus, N- and C-terminal deletions, and 13 amino acid substitutions. Nuclear export of v-ErbA occurs through a CRM1-mediated pathway. In contrast, nuclear export of TRα and another isoform, TRβ, is CRM1-independent. To determine which amino acid changes in v-ErbA confer CRM1-dependent nuclear export, we expressed a panel of green and yellow fluorescent protein-tagged mutant and chimeric proteins in mammalian cells. The sensitivity of subcellular trafficking of these mutants to leptomycin B (LMB), a specific inhibitor of CRM1, was assessed by fluorescence microscopy. Our data showed that a nuclear export sequence resides within a 70-amino acid domain in the C-terminal portion of the p10 region of Gag, and in vitro binding assays demonstrated that Gag interacts directly with CRM1. However, a panel of ligand-binding domain mutants of v-ErbA lacking the Gag sequence exhibited greater nuclear localization in the presence of LMB, suggesting that the various amino acid substitutions/deletions may cause a conformation shift, unmasking an additional CRM1-dependent nuclear export sequence. In contrast, the altered DNA-binding domain of the oncoprotein did not contribute to CRM1-dependent nuclear export. Heterokaryon experiments revealed that v-ErbA did not undergo nucleocytoplasmic shuttling when the CRM1 export pathway was blocked by LMB treatment, suggesting that the ability to follow the export pathway used by TRα has been lost by the oncoprotein during its evolution. Our findings thus point to the intriguing possibility that acquisition of altered nuclear export capabilities contributes to the oncogenic properties of v-ErbA.
Stereotactic Body Radiotherapy as Boost for Organ-confined Prostate Cancer

Stereotactic body radiotherapy (SBRT) boost following external beam radiation therapy (EBRT) for advanced localized prostate cancer may reduce toxicity while escalating the dose. We present preliminary biochemical control and urinary, rectal and sexual toxicities for 73 patients treated with SBRT as a boost to EBRT. Forty-one intermediate- and 32 high-risk localized prostate cancer patients received 45 Gy EBRT with SBRT boost. Twenty-eight patients (38.3%) received a total SBRT boost dose of 18 Gy (3 fractions of 6 Gy), 28 patients (38.3%) received 19.5 Gy (3 fractions of 6.5 Gy), and 17 patients (23.2%) received 21 Gy (3 fractions of 7 Gy). Toxicity was assessed using the Radiation Therapy Oncology Group urinary and rectal toxicity scale. Biochemical failure was assessed using the Phoenix definition. The median follow-up was 33 months (range, 22 − 43 months). Less than 7% Grade II and no higher grade acute toxicities occurred. To date, one Grade III and no Grade IV late toxicities occurred. For the 97% of patients with 24 months minimum follow-up, 71.8% achieved a PSA nadir threshold of 0.5 ng/mL. Three intermediate-risk and seven high-risk biochemical failures occurred; one high-risk patient died of his cancer. Three-year actuarial biochemical control rates were 89.5% and 77.7% for intermediate- and high-risk patients, respectively. SBRT boost for prostate cancer treatment is safe and feasible with minimal acute toxicity. At 33 months late toxicity and biochemical control are promising. Long-term durability of these findings remains to be established.
User Identification for Home Entertainment Based on Free-Air Hand Motion Signatures

A user identification system based on free-air hand signature-gestures acquired with a 3-D camera was developed. In the system, users interactively defined their own motion signatures by demonstration and trained the system by performing a plurality of signatures. The system identifies the user by comparing the distances of a sample to other signatures. The distance metric is learned by using neighborhood components analysis. An interactive enrollment algorithm which uses sequential clustering, and allows the system to advise the user during signature selection and system training was developed. Four validation tests were conducted: 1) all users using a single predefined signature-gesture (independent); 2) each user using a personal signature-gesture (dependent); 3) copycat tests for examining robustness against forgery; and 4) operation of the interactive enrollment system. For identifying a single user out of user cohorts of three to seven people, the independent system had average accuracies of 91%-77% depending on cohort size and signature shape. Higher average accuracies of 98%-92% were obtained for the dependent system. In the forgery tests, users with high signature variability over time were susceptible to forgeries, but users with low signature variability obtained a low equal error rate of 0.083. The interactive enrollment system significantly improved recognition accuracy. The proposed system can be integrated into gesture-based home entertainment systems and used for interface customization, content adaptation, and parental control. User attitudes toward the system within this context were assessed using the widely accepted technology acceptance model. Based on 69 responders, the results indicated a positive user attitude toward the system and a high intention to use it. The users expressed a preference for personalized gestures, a finding that indicates the importance of the interactive enrollment module for personalizing the signature-gestures.
Tele-audiology: Expanding Access to Hearing Care and Enhancing Patient Connectivity.

BACKGROUND Tele-audiology within the Department of Veterans Affairs has expanded significantly in response to growing Veteran hearing needs and issues, namely maldistribution of provider resources and continued lags in timely access to care. Although tele-audiology originated from successive telemedicine programs dating back to the early 1960s, the innovation that is currently being developed broadens tele-audiology and hearing care practice to virtual care modalities such as clinical video telehealth, store-and-forward telehealth, home telehealth, mobile health applications, secure messaging, and electronic consults. PURPOSE The purpose of this article is to address current and emerging tele-audiology initiatives in context of a connected health care practice model. It includes information on initiatives within the Department of Veterans Affairs, provides an overview of current and potential modalities for improving patient hearing care, and identifies the challenges that exist in implementing and sustaining these services. Information is intended to stimulate creative thinking about future innovative service delivery models. CONCLUSION Significant challenges exist in the evolution of innovative tele-audiology services. Despite barriers, these initiatives have the potential to match existing capacity with emerging patient demands in a mutually beneficial partnership and can be successfully integrated into diverse health care and private sector organizations.
Efficient Rejoining of Radiation-induced DNA Double-strand Breaks in Centromeric DNA of Human Cells*

Although major efforts in elucidating different DNA double-strand break (DSB) repair pathways and their contribution to accurate repair or misrepair have been made, little is known about the influence of chromatin structure on the fidelity of DSB repair. Here, the repair of ionizing radiation-induced DSBs was investigated in heterochromatic centromeric regions of human cells in comparison with other genomic locations. A hybridization assay was applied that allows the quantification of correct DSB rejoining events in specific genomic regions by measuring reconstitution of large restriction fragments. We show for two primary fibroblast lines (MRC-5 and 180BR) and an epithelial tumor cell line that restriction fragment reconstitution is considerably more efficient in the centromere than in average genomic locations. Importantly, however, DNA ligase IV-deficient 180BR cells show, compared with repair-proficient MRC-5 cells, impaired restriction fragment reconstitution both in average DNA and in the centromere. Thus, the efficient repair of DSBs in centromeric DNA is dependent on functional non-homologous end joining. It is proposed that the condensed chromatin state in the centromere limits the mobility of break ends and leads to enhanced restriction fragment reconstitution by increasing the probability for rejoining correct break ends.
Maternal mortality in sub‐Saharan Africa: the contribution of ineffective blood transfusion services

In sub‐Saharan Africa, where blood supply is critically inadequate, severe haemorrhage is a leading cause of maternal deaths. The aim of this review was to estimate the impact of lack of blood on maternal deaths and identify reasons and potential solutions. Databases and websites from 1970 to 2007 were searched for information concerning maternal deaths and near misses due to haemorrhage in sub‐Saharan Africa. Original studies that provided qualitative or quantitative information about blood transfusion services in relation to obstetric deaths or near misses were included. Data about maternal haemorrhage deaths associated with lack of blood for transfusion and reasons for blood shortages were extracted from the full text of articles by two independent reviewers using predesigned, piloted forms. Twenty of 37 selected studies described a direct association between maternal deaths and lack of blood transfusions. Five of 37 provided quantitative information which showed that overall 26% (16–72%) of maternal haemorrhage deaths were due to lack of blood. Reasons included nonaffordability of blood, lack of blood donors, unwillingness of relatives to donate and inadequate supplies and transport. Lack of blood for emergency transfusions is a major, but poorly quantified and under‐researched cause of maternal deaths in sub‐Saharan Africa. Potential solutions include more blood donations, better financing mechanisms and more efficient management systems. Complementary approaches to prevent severe anaemia and treat hypovolaemia are important to reduce use of transfusions.