The extracellular domain of herpes simplex virus gE is indispensable for efficient cell-to-cell spread: evidence for gE/gI receptors.
Herpes simplex virus (HSV) spreads rapidly and efficiently within epithelial and neuronal tissues. The HSV glycoprotein heterodimer gE/gI plays a critical role in promoting cell-to-cell spread but does not obviously function during entry of extracellular virus into cells. Thus, gE/gI is an important molecular handle on the poorly understood process of cell-to-cell spread. There was previous evidence that the large extracellular (ET) domains of gE/gI might be important in cell-to-cell spread. First, gE/gI extensively accumulates at cell junctions, consistent with being tethered there. Second, expression of gE/gI in trans interfered with HSV spread between epithelial cells. To directly test whether the gE ET domain was necessary for gE/gI to promote virus spread, a panel of gE mutants with small insertions in the ET domain was constructed. Cell-to-cell spread was reduced when insertions were made within either of two regions, residues 256 to 291 or 348 to 380. There was a strong correlation between loss of cell-to-cell spread function and binding of immunoglobulin. gE ET domain mutants 277, 291, and 348 bound gI, produced mature forms of gE that reached the cell surface, and were incorporated into virions yet produced plaques similar to gE null mutants. Moreover, all three mutants were highly restricted in spread within the corneal epithelium, in the case of mutant 277 to only 4 to 6% of the number of cells compared with wild-type HSV. Therefore, the ET domain of gE is indispensable for efficient cell-to-cell spread. These observations are consistent with our working hypothesis that gE/gI can bind extracellular ligands, so-called gE/gI receptors that are concentrated at epithelial cell junctions. This fits with similarities in structure and function of gE/gI and gD, which is a receptor binding protein.


In vivo monitoring of splenocytes in NMRI nu/nu- and nu/+,- mice during Lewis lung tumor progression or regression.
Splenocytes of T-cell deficient NMRI nu/nu- mice (n=15) and of inbred immunocompetent NMRI nu/+,littermates (n=18) significantly (p<0.05) increased during Lewis lung (LL) tumor growth and eventually decreased. Half of the heterozygous NMRI nu/+,- mice rejected the tumors (LL-regressors). Zymosan-induced and lucigenin-amplified chemiluminescence (CL) of splenic polymorphonuclear leukocytes (PMN) and -precursors significantly (p<0.05) increased in NMRI nu/+,- mice with progressing tumors (LL-progressors) on the 3rd and in LL-regressors and nude mice on the 7th day after tumor inoculation. It subsequently decreased although numbers of PMN/- precursors further multiplied predominantly in the LL-progressors. Another peak of CL was found in those animals after the 20th day. Splenic lymphocytes significantly (p<0.05) increased in all mice from day 5 forward and dropped to original numbers between the 20th and 25th days. Maximal lymphocyte counts were significantly (p<0.05) lower in nude mice as compared to the NMRI nu/+,- littermates. The splenic T-/B-ratio significantly (p<0.005) correlated in LL-regressors and did not in LL-progressors. This correlation could be a suitable marker for tumor progression in NMRI nu/+ mice bearing LL-tumors.


Evaluation of the role of inositol trisphosphate in IgE-dependent exocytosis.
A close correlation exists between inhibition by 12-O-tetradecanoylphorbol 13-acetate (TPA) of inositol trisphosphate (InsP3) formation and the rise in internal Ca2+ concentrations in IgE-stimulated rat basophilic leukemia (RBL-2H3) cells. Inhibition of both processes is dose-dependent, with half-maximal and maximal inhibition occurring at 1.5 and 10 ng of TPA/ml respectively. At a similar range of concentrations TPA does not inhibit, but rather enhances, IgE-dependent secretion. When added to antigen-activated cells. EGTA immediately abrogates secretion and stimulates InsP3 production. In contrast, EGTA has only a small inhibitory effect on IgE-induced secretion from TPA-activated cells. In antigen-activated cells, EGTA partially inhibits InsP1 formation, suggesting that, unlike InsP3, InsP1 may in part be formed directly from phosphatidylinositol in a Ca2(-)-dependent manner. Together, these findings suggest that under physiological conditions the stimulated formation of InsP3 is insufficient for triggering secretion, and that Ca2+ influx is essential. Moreover, InsP3 formation is not obligatory for IgE-mediated exocytosis, provided that the cells are activated by TPA. Secretion from TPA-activated cells, which is independent of InsP3 formation and the rise in internal Ca2+, does not require the presence of external Ca2+, implying that the presence of external Ca2+ during IgE-induced secretion is required for producing the Ca2+ signal and not for exocytosis per se.


Planned randomized conversion from tacrolimus to sirolimus-based immunosuppressive regimen in de novo kidney transplant recipients.
Planned conversion from tacrolimus to sirolimus was evaluated in de novo kidney transplant recipients. In this multicenter, randomized, open-label study, 297 patients were initially treated with tacrolimus, mycophenolate sodium and prednisone. Of the 283 patients reaching 3 months, 97 were converted to sirolimus (SRL), 107 were maintained on tacrolimus (TAC) and 79 were patients receiving TAC without criteria to undergo intervention at month 3 (TACex). The primary objective was to show superior estimated glomerular filtration rate (eGFR) in the SRL group at month 24. Of the 258 patients who completed 24 months, 91 (94%) were in the SRL group, 101 (94%) in the TAC group and 66 (84%) in the TACex group. In the intention-to-treat population there were no differences in eGFR (66.2 ± 25.3 vs. 70.7 ± 25.1, p = 0.817) or in the severity of chronic sclerosing lesions scores in 24-month protocol biopsies. Higher mean urinary protein-to-creatinine ratio (0.36 ± 0.69 vs. 0.15 ± 0.53, p = 0.03) and higher incidence of treated acute rejection between months 3-24 (13.4% vs. 4.7%, p = 0.047) were observed in SRL compared to TAC group. In this population planned conversion from TAC to SRL 3 months after kidney transplantation was not associated with improved renal function at 24 months.


Adsorption characteristics of various proteins on a metal surface in the presence of an external electric potential.
The effect of the properties of a protein on its adsorption to a metal surface in the presence of external electric potential was investigated. Protein adsorption processes at different surface potentials were measured for fifteen types of proteins using an in-situ ellipsometry. The tested proteins were classified into three groups, based on the amount of protein that was adsorbed as a function of the surface potential: In First group of proteins, an increasing trend for the amount adsorbed with a more positive surface potential was found; The amount adsorbed of α-chymotrypsinogen A and ribonuclease A (Second group) were roughly constant and independent of the applied surface electric potentials; In Third group, the amount adsorbed decreased with increasing surface potential. This protein classification was correlated with the isoelectric points of the proteins (First group: ≤9.3; Second group: 9.3-10; Third group: >10). Increasing the pH positively and negatively shifted the surface potentials, allowing ß-lactoglobulin (First group) and lysozyme (Third) to become adsorbed, respectively. The surface potential range for protein adsorption was also markedly shifted depending on the metal substrate type. These findings were interpreted based on the electrostatic interactions among the protein, surface hydroxyl groups, and the applied external electric field.


Parameter convergence and learning curves for neural networks.
We revisit the oft-studied asymptotic (in sample size) behavior of the parameter or weight estimate returned by any member of a large family of neural network training algorithms. By properly accounting for the characteristic property of neural networks that their empirical and generalization errors possess multiple minima, we rigorously establish conditions under which the parameter estimate converges strongly into the set of minima of the generalization error. Convergence of the parameter estimate to a particular value cannot be guaranteed under our assumptions. We then evaluate the asymptotic distribution of the distance between the parameter estimate and its nearest neighbor among the set of minima of the generalization error. Results on this question have appeared numerous times and generally assert asymptotic normality, the conclusion expected from familiar statistical arguments concerned with maximum likelihood estimators. These conclusions are usually reached on the basis of somewhat informal calculations, although we shall see that the situation is somewhat delicate. The preceding results then provide a derivation of learning curves for generalization and empirical errors that leads to bounds on rates of convergence.


Implementing the ICF in Occupational Health; building a curriculum as an exemplary case.
This paper addresses the need for a paradigm shift from post-diagnosis tertiary care towards maintenance and promotion of health across the lifespan, for healthcare in general and in occupational healthcare specifically. It is based on the assumption that the realization of this paradigm shift may be facilitated by teaching (future) occupational health professionals to use the International Classification of Functioning, Disability and Health (ICF). Describing the development of a an ICF based occupational health curriculum. Grafting a training trajectory in the ICF for educating the biopsychosocial health paradigm, onto a training trajectory in the Critical Appraisal of a Topic (CAT), a method for teaching evidence based practice skills. The development process of the training trajectories in the master program Work, Health, and Career at Maastricht University is described as an example of an intervention for shifting the paradigm in healthcare curricula. The expected results are a shift from the biomedical towards the biopsychosocial paradigm, a reductionist approach towards a more holistic view on cases, a reactive way of working towards a more proactive work style, and from using a merely quantifiable evidence base towards using a broad evidence base. Incorporating the biopsychosocial paradigm into the assessment and scientific reasoning skills of students is not only valuable in occupational healthcare but might be a valuable approach for all disciplines in healthcare for which contextual factors are important e.g. rehabilitation, psychiatry and nutritional science.


The Correlation of Serum Myostatin Levels with Gait Speed in Kidney Transplantation Recipients.
The primary role of myostatin is to negatively regulate skeletal muscle growth. The gait speed is a noninvasive, reliable parameter that predicts cardiovascular risk and mortality. This study evaluated the relationship between serum myostatin concentrations and gait speeds in patients who had undergone kidney transplantation (KT). A total of 84 KT recipients were evaluated. A speed of less than 1.0 m/s was categorized into the low gait speed group. We measured serum myostatin concentrations with a commercial enzyme-linked immunosorbent assay. KT recipients in the low gait speed group had significantly older age, as well as higher body weight, body mass index (BMI), skeletal muscle index, serum triglyceride levels, glucose levels, and blood urea nitrogen levels, lower estimated glomerular filtration rates and serum myostatin levels, a higher percentage of steroid use, and a lower proportion of mycophenolate mofetil use. Multivariable logistic regression analysis revealed that lower myostatin levels and lower frequency of mycophenolate mofetil use were independently associated with low gait speed. In multivariable stepwise linear regression analysis, myostatin levels were positively correlated with gait speeds, and age and BMI were negatively correlated with gait speeds. In the study, serum myostatin levels were significantly lower in the low gait speed group. Subjects in the low gait speed group also had greater BMI and older age.


[An operated case of bitemporal lobe epilepsy associated with old infarction in the left occipital lobe].
A 15-year-old female, who presented with bitemporal lobe epilepsy associated with old infarction in the left occipital lobe, was reported. MRI with fluid attenuated inversion recovery sequence demonstrates cortical atrophy with hyperintensity of the white matter in the left occipital lobe as well as volume loss and hyperintensity of the left hippocampus. Interictal positron emission tomography with [18F]fluorodeoxy glucose (FDG-PET) and single photon emission computed tomography with technetium-99m-ethyl-cysteinate dimer indicate hypometabolism and hypoperfusion in the left occipital lobe and the left temporal lobe, respectively. Scalp recorded EEG did not lateralize the side of the epileptogenic zone. Chronic subdural electrode recording demonstrated that the ictal onset zones were located in the bilateral side of the temporal lobe. Eighty-nine percent of 19 spontaneous seizures were left sided onset. The anterior temporal lobectomy with hippocampectomy was performed for the left side. Although temporal lobe epilepsy is sometimes a bilateral disease, unilateral lobectomy for a strong predominant side, based on the MRI and FDG-PET findings, is effective for some patients.


[Assessment of a postural reference frame as a diagnostic test for lumbar lordotic posture].
Studying lumbar lordosis (LL) in large patient populations is not a practical approach due to the technical limitations of the gold standard. Determine the specificity of a postural reference frame (PRF) in the diagnosis of LL. A PRF (wooden frame using strings as a reference for measuring the distance between lumbar curvature and the glutei) was used as a diagnostic test to asses LL. The Ferguson angle, a radiologic evaluation, was used as gold standard. Specificity, sensitivity, positive predictive value (PPV), and negative predictive value (NPV) of the PRF were determined. One hundred and ten subjects were studied (58 women, 52 men). Mean age was 18.8 +/- 3 years, weight 63 +/- 12 kg, height 164 +/- 8 cm and body mass index (BMI) 23.2 +/- 4 kg/m2. With a cut-off point 37 cm, test specificity was 94%, sensitiivity was 17%, PPV was 50%, and NPV was 75%. The postural reference frame as an assessment tool is highly specific and can aid in the diagnosis of LL among young asymptomatic patients.


Development of a prognostic model for six-month mortality in older adults with declining health.
Estimation of six-month prognosis is essential in hospice referral decisions, but accurate, evidence-based tools to assist in this task are lacking. To develop a new prognostic model, the Patient-Reported Outcome Mortality Prediction Tool (PROMPT), for six-month mortality in community-dwelling elderly patients. We used data from the Medicare Health Outcomes Survey linked to vital status information. Respondents were 65 years old or older, with self-reported declining health over the past year (n=21,870), identified from four Medicare Health Outcomes Survey cohorts (1998-2000, 1999-2001, 2000-2002, and 2001-2003). A logistic regression model was derived to predict six-month mortality, using sociodemographic characteristics, comorbidities, and health-related quality of life (HRQOL), ascertained by measures of activities of daily living and the Medical Outcomes Study Short Form-36 Health Survey; k-fold cross-validation was used to evaluate model performance, which was compared with existing prognostic tools. The PROMPT incorporated 11 variables, including four HRQOL domains: general health perceptions, activities of daily living, social functioning, and energy/fatigue. The model demonstrated good discrimination (c-statistic=0.75) and calibration. Overall diagnostic accuracy was superior to existing tools. At cut points of 10%-70%, estimated six-month mortality risk sensitivity and specificity ranged from 0.8% to 83.4% and 51.1% to 99.9%, respectively, and positive likelihood ratios at all mortality risk cut points ≥40% exceeded 5.0. Corresponding positive and negative predictive values were 23.1%-64.1% and 85.3%-94.5%. Over 50% of patients with estimated six-month mortality risk ≥30% died within 12 months. The PROMPT, a new prognostic model incorporating HRQOL, demonstrates promising performance and potential value for hospice referral decisions. More work is needed to evaluate the model.


From science student to scientist: Predictors and outcomes of heterogeneous science identity trajectories in college.
This 5-year longitudinal study investigates the development of science identity throughout college from an expectancy-value perspective. Specifically, heterogeneous developmental patterns of science identity across 4 years of college were examined using growth-mixture modeling. Gender, race/ethnicity, and competence beliefs (efficacy for science tasks, perceived competence in science) were modeled as antecedents, and participation in a science career after graduation was modeled as a distal outcome of these identity development trajectories. Three latent classes (High with Transitory Incline, Moderate-High and Stable, and Moderate-Low with Early Decline) were identified. Gender, race/ethnicity, and competence beliefs in the first year of college significantly predicted latent class membership. In addition, students in the two highest classes were significantly more likely to report being involved in science careers or science fields after college graduation than students in the Moderate-Low with Early Decline class. (PsycINFO Database Record


Development and application of a microarray meter tool to optimize microarray experiments.
Successful microarray experimentation requires a complex interplay between the slide chemistry, the printing pins, the nucleic acid probes and targets, and the hybridization milieu. Optimization of these parameters and a careful evaluation of emerging slide chemistries are a prerequisite to any large scale array fabrication effort. We have developed a 'microarray meter' tool which assesses the inherent variations associated with microarray measurement prior to embarking on large scale projects. The microarray meter consists of nucleic acid targets (reference and dynamic range control) and probe components. Different plate designs containing identical probe material were formulated to accommodate different robotic and pin designs. We examined the variability in probe quality and quantity (as judged by the amount of DNA printed and remaining post-hybridization) using three robots equipped with capillary printing pins. The generation of microarray data with minimal variation requires consistent quality control of the (DNA microarray) manufacturing and experimental processes. Spot reproducibility is a measure primarily of the variations associated with printing. The microarray meter assesses array quality by measuring the DNA content for every feature. It provides a post-hybridization analysis of array quality by scoring probe performance using three metrics, a) a measure of variability in the signal intensities, b) a measure of the signal dynamic range and c) a measure of variability of the spot morphologies.


Experiences of gout-related disability from the patients' perspective: a mixed methods study.
Disability is a common problem in patients with gout. Recently, the Health Assessment Questionnaire Disability Index (HAQ-DI) was recommended for assessing patient reported activity limitations in gout. However, few studies have explored experiences and issues of disability from the perspective of gout patients themselves. The objectives of this study were to explore disability issues in patients with gout and to examine the content validity of the HAQ-DI in this patient group. Two studies were performed: a semi-structured interview study with seven male gout patients and a cross-sectional survey study among 34 other patients with gout. In both studies, specific examples of activity limitations were elicited using open-ended methods. The survey study additionally aimed to quantify the relevance of the activities listed in the HAQ-DI. Most patients experienced several gout attacks in the previous year. Limitations were reported to occur during a flare, but patients were generally not limited between attacks. During an attack, patients mainly experienced limitations related to mobility, especially walking and climbing stairs. Patients also mentioned limitations in activities related to domestic life, such as gardening and doing housework. Limitations related to self-care or activities requiring the use of the upper extremities were rarely mentioned. Corresponding HAQ-DI items were skewed towards very low disability scores over the past week. Assessments of gout-related disability should particularly focus on mobility and lower extremity functioning and should consider the intermittent nature of the disease. The HAQ-DI may not adequately meet these requirements, suggesting the need to explore other measures of gout-related disability


Factors associated with injuries in adolescents, from the National Adolescent School-based Health Survey (PeNSE 2012).
To estimate the prevalence of injuries among teenagers and to examine the associated risk factors, such as sociodemographic characteristics, risk behaviors, family ties and other factors. The prevalence of the outcome (injury) was estimated with a 95%confidence interval. In order to verify factors associated with the injury, a bivariate analysis was made with estimated odds ratio (OR) and its respective confidence intervals. Then, a multivariate analysis was carried out, only with variables whose descriptive level was equal to or lower than 5% (p < 0.05) remaining in the model. The study of injury in adolescents, based on the data from the National Adolescent School-based Health Survey (PeNSE), pointed out that 10.3% of the teenagers suffered severe injuries in the past 12 months, such as cuts or perforations, broken bones or dislocated joints. The following variables remained independently associated with "suffering severe injuries": being a male teenager; black, mulatto or indigenous race/color and working. Factors related to family ties are significant when the relations are fragile amongst members: adolescents that are injured the most are the ones who suffer most aggressions at home, who skip classes without notifying their parents, those who do not live with their parents and have low family control. The most relevant aspects of mental health are insomnia and loneliness. The factors associated to the exposure to situations of violence that remained in the model were: insecurity in school and in the route home-school; getting a ride with someone inebriated; drinking and driving motorized vehicles; not wearing the seatbelt; not wearing a helmet and being bullied. Among the factors of individual behavior, the following can be emphasized: use of alcohol, cigarettes, trying illicit drugs and early sexual intercourse. The analysis of the determinants for suffering injuries in childhood and adolescence shows the complex relationship between associated factors, which points to the need for action towards several aspects to reduce social inequalities, strengthen family ties and prevent violent contexts and individual risk factors.


Case report: DSP truncation variant p. R1951X leads to arrhythmogenic left ventricular cardiomyopathy.
Standardized diagnostic criteria for arrhythmogenic left ventricular cardiomyopathy (ALVC) have been recently proposed. The criteria emphasize structural left ventricle (LV) myocardial change on contrast-enhanced imaging and require the identification of gene variants associated with arrhythmogenic cardiomyopathy. A 21-year-old man presented for evaluation of exertional syncope and was found to have monomorphic ventricular tachycardia (VT) and an episode of polymorphic VT that degenerated to ventricular fibrillatory cardiac arrest. Documented premature ventricular contractions were of left bundle branch block, inferior axis morphology. Ventricular arrhythmias were successfully suppressed with β-blockade, amiodarone, and lidocaine, and a subcutaneous implantable cardioverter-defibrillator was implanted. Cardiac magnetic resonance imaging demonstrated normal-appearing right ventricle, reduced LV ejection fraction, and sub-epicardial scarring of basal-anterior and anterolateral LV segments. Endomyocardial biopsy showed lymphocytic myocarditis, and genetic testing revealed a pathogenic truncating mutation in the DSP gene, which encodes desmoplakin; this variant was also identified in the patient's mother who carried a diagnosis of non-ischaemic cardiomyopathy. These findings are consistent with a diagnosis of ALVC. The clinical presentation of ALVC can be very dramatic. The differential for sub-epicardial LV myocardial fibrosis includes myocarditis, sarcoidosis, and in those with a suspicious family history or characteristic electrocardiogram findings, genetic cardiomyopathy. Prompt referral to a genetic counsellor can be lifesaving to patients and their family members.


Language-tree divergence times support the Anatolian theory of Indo-European origin.
Languages, like genes, provide vital clues about human history. The origin of the Indo-European language family is "the most intensively studied, yet still most recalcitrant, problem of historical linguistics". Numerous genetic studies of Indo-European origins have also produced inconclusive results. Here we analyse linguistic data using computational methods derived from evolutionary biology. We test two theories of Indo-European origin: the 'Kurgan expansion' and the 'Anatolian farming' hypotheses. The Kurgan theory centres on possible archaeological evidence for an expansion into Europe and the Near East by Kurgan horsemen beginning in the sixth millennium BP. In contrast, the Anatolian theory claims that Indo-European languages expanded with the spread of agriculture from Anatolia around 8,000-9,500 years bp. In striking agreement with the Anatolian hypothesis, our analysis of a matrix of 87 languages with 2,449 lexical items produced an estimated age range for the initial Indo-European divergence of between 7,800 and 9,800 years bp. These results were robust to changes in coding procedures, calibration points, rooting of the trees and priors in the bayesian analysis.


Safety education in a pediatric primary care setting.
Parents of 171 children coming to the Yale-New Haven Hospital Primary Care Center for their 6-month checkup were randomized into an intervention group (n = 85) and a control group (n = 86). Parents in the intervention group received a three-part individualized course in child safety that required active parental participation. Parts 1, 2, and 3 were given at the 6-month, 9-month, and 12-month well-child visits, respectively. Parents in the control group received routine safety education as provided at well-child visits. The educational phase of the study was completed by 129 families, 65 in the intervention group and 64 in the control group. Safety knowledge, number of hazards in the home, and reported accidents were assessed by a "blinded" community health worker approximately 1 month after the 12-month well-child visit. A total of 109 home visits were made, 55 for the intervention group and 54 for the control group. Parental safety knowledge was assessed based upon pictorial hazard recognition. Of 13 possible hazards, the mean number of hazards recognized by the intervention group parents was 9.4 (n = 55) v 8.4 (n = 50) by the control group parents (t = 2.1, P less than .05, two-tailed). A hazard score was determined for each family based on nine possible hazards observed at the home visit. The mean hazard score for the intervention group was 2.4 (n = 55 v 3.0 (n = 54) for the control group (t = 2.4, P less than .02, two-tailed). Parentally reported accidents and accidents reported in hospital records were similar for both groups. Results of this study suggest that age-appropriate safety education that is repetitive and individualized and that requires active parental participation results in an increase in parental knowledge and an improvement in certain safety practices.


An overview of the California workers' compensation system.
Nearly every working Californian is protected by workers' compensation for any job-related injury. Because of its size and the nature of its program, California pays out twice as much in benefits as any other state. The author describes the administration of the compensation system, its costs and the results.


Psychotherapy as adjunct in treatment of vomiting during pregnancy.
Adjunctive psychotherapeutic measures were used to treat ten consecutive pregnant women referred to a psychiatric consultation service because of severe intractable vomiting. In addition to customary medical measures, they received three types of psychotherapy: (1) supportive psychotherapy (establishment of positive relationship, frequent reassuring conversations, and encouragement of expression of thoughts and feelings); (2) hypnotherapy (trance induction and suggestions of comfort in the gastrointestinal tract, desirability of feeling of substance in stomach, and ability to retain and digest food eaten); and (3) behavior modification (positive reinforcement for retaining food and gaining weight, through granting desirable considerations such as visitors, radio, television, and being up in or outside the room). Results were encouraging. Nine patients recovered and completed normal pregnancies, while one improved but later aborted due to other complications. When compared with a control group of routinely treated patients, recovery was more rapid.


Bond length alternation and energy band gap of polyyne.
The bond length alternation (BLA) and energy band gap of polyyne are investigated by various first-principles theories, including Hartree-Fock, MP2, hybrid, and nonhybrid density functional theories. Both solid-state calculations utilizing periodic boundary conditions on polymers and molecular quantum mechanical calculations on extra-long oligomers were performed with consistent results. By validation on similar linear conjugated polymers, polyacetylene and polydiacetylene, the combination of hybrid-DFT schemes, B3LYP//BHandHLYP or B3LYP//KMLYP, is shown to give the best predictions for both geometry and band gap of polyyne based on available experimental data. We conclude that the best estimate of the BLA of polyyne is about 0.13 A and that of the band gap is about 2.2 eV.


Highly sensitive mutation quantification by high-dynamic-range capillary-array electrophoresis (HiDy CE).
A simple and robust ultra-small four-color-fluorescence detection system was developed by integrating its components, namely, a four-capillary array, an injection-molded-plastic four-lens array, a four-dichroic-mirror array, and a CMOS sensor, as one device. The developed system was applied to a high-dynamic-range capillary-array electrophoresis (HiDy CE) to quantify a rare EGFR mutant (MT) of exon 19 deletion in a large excess of EGFR wild type (WT). Samples with serially diluted MT and constant-concentration WT were co-amplified by competitive PCR and subjected to HiDy CE. The MT peak in each electropherogram was then compared to the WT peak. As a result, MT was quantified with high-sensitivity (LOD of 0.004% MT/WT) and four-orders-of-magnitude dynamic range (0.01-100% MT/WT) by HiDy CE. Moreover, compared with existing methods, HiDy CE achieves higher speed, higher sample throughput, and lower consumable cost per sample. It has therefore great potential to be used in clinical practice.


Mapping the binding site of colchicinoids on beta -tubulin. 2-Chloroacetyl-2-demethylthiocolchicine covalently reacts predominantly with cysteine 239 and secondarily with cysteine 354.
2-Chloroacetyl-2-demethylthiocolchicine (2CTC) and 3-chloroacetyl-3-demethylthiocolchicine (3CTC) resemble colchicine in binding to tubulin and react covalently with beta-tubulin, forming adducts with cysteine residues 239 and 354. The adducts at Cys-239 are less stable than those at Cys-354 during formic acid digestion. Extrapolating to zero time, the Cys-239 to Cys-354 adduct ratio is 77:23 for 2CTC and 27:73 for 3CTC. Using energy minimization modeling to dock colchicinoids into the electron crystallographic model of beta-tubulin in protofilaments (Nogales, E. , Wolf, S. G., and Downing, K. H. (1998) Nature 391, 199-203), we found two potential binding sites. At one, entirely encompassed within beta-tubulin, the C2- and C3-oxygen atoms of 2CTC and 3CTC overlapped poorly with those of colchicine and thiocolchicine, but distances from the reactive carbon atoms of the analogs to the sulfur atoms of the cysteine residues were qualitatively consistent with reactivity. The other potential binding site was located at the alpha/beta interface. Here, the oxygen atoms of the analogs overlapped well with those of colchicine, but relative distances of the reactive carbons to the cysteine sulfur atoms did not correlate with the observed reactivity. A significant conformational change must occur in the colchicine binding site of tubulin in the transition from the unpolymerized to the polymerized state.


Acute ethanol modulation of neurocircuit function in the nucleus of the tractus solitarius.
The nucleus of the tractus solitarius (NTS) is a brain stem region critical to many physiologic processes and has been implicated in addiction to multiple classes of abused drugs, including alcohol (EtOH). That said, the mechanism by which EtOH modulates NTS neurocircuit activity is not well characterized and has yet to be examined utilizing electrophysiologic methods in mouse models of alcohol use disorders. To begin to address this gap in knowledge, we sought to use whole-cell and cell-attached recordings to determine the mechanism of acute EtOH action on GABAergic and glutamatergic neurotransmission, as well as on action potential firing in the NTS of adult male, EtOH naïve mice. Bath application of EtOH (50mM) significantly enhanced the frequency of spontaneous inhibitory postsynaptic current events, while increasing the amplitude of these events in half of the neurons tested. This finding suggests a presynaptic mechanism of EtOH action on GABAergic transmission in the NTS as well as a postsynaptic mechanism in subsets of NTS neurons. EtOH application was further associated with a significant decrease in action potential firing in most, but not all, NTS neurons tested. EtOH induced a small but significant decrease in spontaneous excitatory postsynaptic current frequency, indicating that EtOH may also inhibit NTS glutamatergic signaling to some degree. Intriguingly, in vivo EtOH exposure (4g/kg IP) enhanced c-FOS colocalization with tyrosine hydroxylase via immunohistochemical methods, indicating that NTS norepinephrine neurons may be activated by acute EtOH exposure. Although future work is needed, the current data indicate that acute EtOH may enhance GABAergic signaling in local NTS circuits resulting in disinhibition of NTS norepinephrine neurons. Such a finding has important implications in understanding the role of the NTS in the development of alcoholism.


Cognitive and school functioning in children and adolescents with chronic pain: a critical review.
Cognitive function is a critical factor related to a child's overall developmental trajectory. There is increasing evidence that chronic pain disrupts cognitive function in adults. Little is known about the nature or impact of cognitive disruption in children and adolescents with chronic pain. The present review examines the current literature related to cognitive function in children and adolescents with chronic pain, implications of these findings and future research directions. Nine studies on this topic were found, with a relatively recent increase in publications related to school attendance and subjective studies of school performance. The studies that were found on this topic suggested that chronic pain affects cognitive function in children but the scope of these effects on children's function and developmental trajectories is not yet clear. While methodological issues surely make it difficult to study cognitive function in children with chronic pain, the potential gains from such research warrant a pursuit of such work. Much remains to be studied on this important topic.


[Set of sterilization equipment for large therapeutic institutions].
Some problems concerning the organization of sterilization departments in medical and prophylactic institutions of the Health Ministry of the USSR are discussed. A fundamentally new organizational scheme of work according to the three-zone principle is described. The set of equipment for large sterilization departments is presented. It ensures the operation based on the three-zone system.


Protection against myocardial infarction and no-reflow through preservation of vascular integrity by angiopoietin-like 4.
Increased permeability, predominantly controlled by endothelial junction stability, is an early event in the deterioration of vascular integrity in ischemic disorders. Hemorrhage, edema, and inflammation are the main features of reperfusion injuries, as observed in acute myocardial infarction (AMI). Thus, preservation of vascular integrity is fundamental in ischemic heart disease. Angiopoietins are pivotal modulators of cell-cell junctions and vascular integrity. We hypothesized that hypoxic induction of angiopoietin-like protein 4 (ANGPTL4) might modulate vascular damage, infarct size, and no-reflow during AMI. We showed that vascular permeability, hemorrhage, edema, inflammation, and infarct severity were increased in angptl4-deficient mice. We determined that decrease in vascular endothelial growth factor receptor 2 (VEGFR2) and VE-cadherin expression and increase in Src kinase phosphorylation downstream of VEGFR2 were accentuated after ischemia-reperfusion in the coronary microcirculation of angptl4-deficient mice. Both events led to altered VEGFR2/VE-cadherin complexes and to disrupted adherens junctions in the endothelial cells of angptl4-deficient mice that correlated with increased no-reflow. In vivo injection of recombinant human ANGPTL4 protected VEGF-driven dissociation of the VEGFR2/VE-cadherin complex, reduced myocardial infarct size, and the extent of no-reflow in mice and rabbits. These data showed that ANGPTL4 might constitute a relevant target for therapeutic vasculoprotection aimed at counteracting the effects of VEGF, thus being crucial for preventing no-reflow and conferring secondary cardioprotection during AMI.


Cascaded redundancy reduction.
We describe a method for incrementally constructing a hierarchical generative model of an ensemble of binary data vectors. The model is composed of stochastic, binary, logistic units. Hidden units are added to the model one at a time with the goal of minimizing the information required to describe the data vectors using the model. In addition to the top-down generative weights that define the model, there are bottom-up recognition weights that determine the binary states of the hidden units given a data vector. Even though the stochastic generative model can produce each data vector in many ways, the recognition model is forced to pick just one of these ways. The recognition model therefore underestimates the ability of the generative model to predict the data, but this underestimation greatly simplifies the process of searching for the generative and recognition weights of a new hidden unit.


Enzymatic assay of beta-lactamase using circular dichroism spectropolarimetry.
A method for measuring the rates of enzymatic hydrolysis of beta-lactam antibiotics based on circular dichroism spectropolarimetry is described. Unhydrolyzed beta-lactam antibiotics have high molar ellipticities, but the hydrolyzed compounds are circular dichroism (CD) inactive in the case of penams or have significantly different CD spectra in the case of cephems. By measuring CD at constant wavelength as a function of time for reaction mixtures containing beta-lactamase and beta-lactam antibiotics, rates of hydrolysis and steady-state enzyme kinetic constants can be derived. The method was applied to measurement of a wide range of enzymatic reaction constants for wild-type and four mutant RTEM-1 beta-lactamases. Compared to the commonly employed assay based on ultraviolet spectroscopy, the new method offers several advantages. These include the ability to measure larger enzymatic Michaelis-Menten constants, less interference from high concentrations of beta-lactamase, higher sensitivity, and potentially less interference from other uv-absorbing components of complex reaction mixtures.


Newer diagnostic modalities for tuberculosis.
The gold standard for diagnosis of tuberculosis is demonstration of mycobacteria from various body fluids. This is often not possible in children due to pauci-bacillary nature of illness. Significant improvement in understanding of molecular biology of Mycobacterium tuberculosis has led to development of newer diagnostic techniques of tuberculosis. Polymerase chain reaction (PCR) is an emerging diagnostic tool for diagnosis of TB in children. However, its role in day-to-day clinical practice needs to be defined. A negative PCR never eliminates possibility of tuberculosis, and a positive result is not always confirmatory. The PCR may be useful in evaluating children with significant pulmonary disease when diagnosis is not readily established by other means, and in evaluating immunocompromised children (HIV infection) with pulmonary disease. In the absence of good diagnostic methods for tuberculosis, a lot of interest has been generated in serodiagnosis. ELISA has been used to detect antibodies to various purified or complex antigens of M. tuberculosis in children. Despite a large number of studies published over the past several years, serology has found little place in the routine diagnosis of tuberculosis in children, even though it is rapid and does not require specimen from the site of disease. Sensitivity and specificity depend on the antigen used, gold standard for the diagnosis of tuberculosis and the type of tubercular infection. Though most of these tests have high specificity, their sensitivity is poor. In addition, these tests may be influenced by factors such as age, prior BCG vaccination and exposure to environmental mycobacteria. The serological tests, theoretically, may not be able to differentiate between infection and disease. At present, serodiagnosis does not appear to have any role in diagnosis of childhood pulmonary tuberculosis. A new test (QuantiFERON-TB or QFT) that measures the release of interferon-gamma in whole blood in response to stimulation by purified protein derivative is comparable with the tuberculin skin testing to detect latent tubercular infection, and is less affected by BCG vaccination. It can also discriminate responses due to nontuberculous mycobacteria, and avoids variability and subjectivity associated with placing and reading the tuberculin skin test. Polymerase chain reaction based test for identification of katG and rpoB mutation which are associated with isoniazid and rifampicin resistance may help in early identification of drug resistance in mycobacterium.


Suboptimal choices and dosing of statins at start of therapy.
To assess dosing and determinants of the choice of statins among starters of statins. Data were obtained from the PHARMO database comprising pharmacy and linked hospital discharge records of approximately 300 000 subjects in the Netherlands. All new users of statins in 1998 were selected. Patient characteristics and drug regimens were compared between starters of different statins. Odds ratios (OR) and 95% confidence intervals (CI) were calculated using polytomous logistic regression modelling, using the start of simvastatin therapy as reference category. In 1998, 1738 patients started using simvastatin (41.1%), pravastatin (23.1%), fluvastatin (11.9%), atorvastatin (22.8%) or cerivastatin (1.0%). Compared with starters with simvastatin [mean dose 1.02 +/- 0.39 defined daily doses (DDDs)], starters with pravastatin (1.27 +/- 0.56 DDDs) and atorvastatin (1.43 +/- 0.59 DDDs) received higher doses (P < 0.001), whereas users of fluvastatin (0.78 +/- 0.37 DDDs) and cerivastatin (0.81 +/- 0.30 DDDs) received lower doses (P < 0.001). Patients already using CYP3A4 inhibitors more frequently received fluvastatin (OR = 1.80; 95% CI 1.11, 2.94), metabolized by non-CYP3A4 pathways, and atorvastatin (OR = 1.62; 95% CI 1.06, 2.47), which is metabolized by CYP3A4, than simvastatin. Statin doses were not adjusted when prescribed to patients using CYP3A4 inhibitors. Many patients starting statin therapy did not receive a statin of first choice. The coadministration of potentially interacting drugs may have led to a change in statin choice, but not in dosage lowering. These findings suggest that the quality of statin therapy could be improved.


Psychometric evaluation of the FACT Colorectal Cancer Symptom Index (FCSI-9): reliability, validity, responsiveness, and clinical meaningfulness.
Patient-reported outcomes (PROs) are essential for evaluating treatment effects on health-related quality of life and symptoms from the patient's perspective. This study sought to evaluate the psychometric properties of the nine-item Functional Assessment of Cancer Therapy/National Comprehensive Cancer Network Colorectal Cancer Symptom Index (FCSI-9) in a metastatic colorectal cancer (mCRC) population. The FCSI-9 and EQ-5D were administered every 2-4 weeks to mCRC subjects in a phase III clinical trial. Three hundred ninety-one mCRC subjects completed the questionnaires at baseline and at least one follow-up assessment. Internal consistency reliability, test-retest reliability, construct validity, known groups validity, responsiveness, and the minimum important difference (MID) of the FCSI-9 were evaluated. The internal consistency and test-retest reliability of the FCSI-9 were acceptable (0.81 and 0.76, respectively). Construct validity was supported based on moderate correlations with the EQ-5D. Known groups validity was evaluated by examining the FCSI-9 scores of subjects categorized by their Eastern Cooperative Oncology Group performance status (PS) score. Subjects with better PS scores reported significantly higher FCSI-9 scores than those with lower PS scores at both baseline and week 8. Responsiveness, as measured by Guyatt's statistic, was 0.77 from baseline to week 8 and 0.60 from week 4 to week 12. Considering all data together, the MID of the FCSI-9 is estimated to be in the range of 1.5-3.0 points. Results provide preliminary evidence of the reliability, validity, and responsiveness of the FCSI-9.


Sustainable development goals for global health: facilitating good governance in a complex environment.
Increasing complexity is following in the wake of rampant globalization. Thus, the discussion about Sustainable Development Goals (SDGs) requires new thinking that departs from a critique of current policy tools in exploration of a complexity-friendly approach. This article argues that potential SDGs should: treat stakeholders, like states, business and civil society actors, as agents on different aggregate levels of networks; incorporate good governance processes that facilitate early involvement of relevant resources, as well as equitable participation, consultative processes, and regular policy and programme implementation reviews; anchor adoption and enforcement of such rules to democratic processes in accountable organizations; and include comprehensive systems evaluations, including procedural indicators. A global framework convention for health could be a suitable instrument for handling some of the challenges related to the governance of a complex environment. It could structure and legitimize government involvement, engage stakeholders, arrange deliberation and decision-making processes with due participation and regular policy review, and define minimum standards for health services. A monitoring scheme could ensure that agents in networks comply according to whole-systems targets, locally defined outcome indicators, and process indicators, thus resolving the paradox of government control vs. local policy space. A convention could thus exploit the energy created in the encounter between civil society, international organizations and national authorities.


KPU-300, a Novel Benzophenone-Diketopiperazine-Type Anti-Microtubule Agent with a 2-Pyridyl Structure, Is a Potent Radiosensitizer That Synchronizes the Cell Cycle in Early M Phase.
KPU-300 is a novel colchicine-type anti-microtubule agent derived from plinabulin (NPI-2358). We characterized the effects of KPU-300 on cell cycle kinetics and radiosensitization using HeLa cells expressing the fluorescent ubiquitination-based cell cycle indicator (Fucci). Cells treated with 30 nM KPU-300 for 24 h were efficiently synchronized in M phase and contained clearly detectable abnormal Fucci fluorescence. Two-dimensional flow-cytometric analysis revealed a fraction of cells distinct from the normal Fucci fluorescence pattern. Most of these cells were positive for an M phase marker, the phosphorylated form of histone H3. Cells growing in spheroids responded similarly to the drug, and the inner quiescent fraction also responded after recruitment to the growth fraction. When such drug-treated cells were irradiated in monolayer, a remarkable radiosensitization was observed. To determine whether this radiosensitization was truly due to the synchronization in M phase, we compared the radiosensitivity of cells synchronized by KPU-300 treatment and cells in early M phase isolated by a combined method that took advantage of shake-off and the properties of the Fucci system. Following normalization against the surviving fraction of cells treated with KPU-300 alone, the surviving fractions of cells irradiated in early M phase coincided. Taken together with potential vascular disrupting function in vivo, we propose a novel radiosensitizing strategy using KPU-300.


Effect of Pharmacoprophylaxis on Postoperative Outcomes in Adult Elective Colorectal Surgery: A Multi-Center Retrospective Cohort Study within an Enhanced Recovery after Surgery Framework.
The application of enhanced recovery after surgery principles decreases postoperative complications (POCs), length of stay (LOS), and readmissions. Pharmacoprophylaxis decreases morbidity, but the effect of specific regimens on clinical outcomes is unclear. Records of 476 randomly selected adult patients who underwent elective colorectal surgeries (ECRS) at 10 US hospitals were abstracted. Primary outcomes were surgical site infection (SSI), venous thromboembolism (VTE), postoperative nausea and vomiting (PONV), pain, and ileus rates. Secondary outcomes included LOS and 7- and 30-day readmission rates. POC rates were SSI (3.4%), VTE (1.5%), PONV (47.9%), pain (58.1%), and ileus (16.1%). Cefazolin 2 g/metronidazole 500 mg and ertapenem 1 g were associated with the shortest LOS; cefotetan 2 g and cefoxitin 2 g with the longest LOS. No SSI occurred with ertapenem and cefotetan. More Caucasians than Blacks received oral antibiotics before intravenous antibiotics without impact. Enoxaparin 40 mg subcutaneously daily was the most common inpatient and discharge VTE prophylaxis. All in-hospital VTEs occurred with unfractionated heparin. Most received rescue rather than around-the-clock antiemetics. Scopolamine patches, spinal opioids, and IV lidocaine continuous infusion were associated with lower PONV. Transversus abdominis plane block with long-acting local anesthetics, celecoxib, non-anesthetic ketamine bolus, ketorolac IV, lidocaine IV, and pregabalin were associated with lower in-hospital pain severity rates. Gabapentinoids and alvimopan were associated with lower ileus rates. Acetaminophen, alvimopan, famotidine, and lidocaine patches were associated with shorter LOS. Significant differences in pharmacotherapy regimens that may improve primary and secondary outcomes in ECRS were identified. In adult ECRS, cefotetan or ertapenem may be better regimens for preventing in-hospital SSI, while ertapenem or C/M may lead to shorter LOS. The value of OA to prevent SSI was not demonstrated. Inpatient enoxaparin, compared to UFH, may reduce VTE rates with a similar LOS. A minority of patients had a documented PONV risk assessment, and a majority used as-needed rather than around-the-clock strategies. Preoperative scopolamine patches continued postoperatively may lower PONV and PDNV severity and shorter LOS. Alvimopan may reduce ileus and shorten LOS. Anesthesia that includes TAP block, ketorolac IV, and pregabalin use may lead to reduced pain rates. Acetaminophen, alvimopan, famotidine, and lidocaine patches may shorten LOS. Given the challenges of pain management and the incidence of PONV/PDNV found in this study, additional studies should be conducted to determine optimal opioid-free anesthesia and the benefit of newer antiemetics on patient outcomes. Moreover, future research should identify latent pharmacotherapy variables that impact patient outcomes, correlate pertinent laboratory results, and examine the impact of order or care sets used for ECRS at study hospitals.


Total parenteral nutrition.
Total parenteral nutrition (TPN) is a relatively new innovation in patient care which allows us to replace and maintain essential nutrients in patients in whom oral or tube feedings are contraindicated or inadequate. Insertion of a catheter into a large central vein permits one to concentrate hypertonic dextrose calories in normal daily fluid requirements. In addition, TPN solutions contain synthetic amino acids or protein hydrolysates, macroelements, electrolytes, and vitamins. Indications for TPN include intestinal fistulas, severe short bowel syndrome, unresolving pancreatitis, advanced inflammatory bowel disease, delayed postoperative gastrointestinal function, developmental anomalies of the intestinal tract, protracted diarrhea of infancy, and hypermetabolic states. Complications encountered in patients receiving TPN are catheter-related mechanical problems, infections, and metabolic abnormalities. In select patients, who otherwise would require repeated hospitalizations for malnutrition, encouraging results have been achieved by the use of TPN in the home.


In-frame deletion in FLNA causing familial periventricular heterotopia with skeletal dysplasia in males.
Periventricular heterotopia (PH) is an etiologically heterogeneous disorder characterized by nodules of neurons ectopically placed along the lateral ventricles. Truncating and missense mutations of the FLNA gene have been identified in almost 100% of families and 26% of sporadic patients with PH. The otopalatodigital syndrome spectrum is caused by distinct FLNA missense mutations or in-frame deletions disrupting the development of craniofacial and long bones. We report on a clinical, neuroimaging, X-ray, and molecular study of a family in which classical bilateral PH appeared as an isolated anatomic feature in the mother and was associated with skeletal abnormalities and facial dysmorphisms in her two sons. Both boys exhibited PH associated with flat face and spatulate finger tips, short broad phalanx and metacarpus, and bowed radius with dislocated wrist joints. All three patients harbored the c.7865_7870del in-frame deletion (p.2622_2623delDK) in the carboxyl-terminal domain (repeat 24) of FLNA. The X-inactivation observed in the mother was skewed towards the mutant allele, resulting in the preferential expression of the wild-type allele. The in-frame deletion in the carboxyl-terminal domain of FLNA caused a phenotype in which PH was associated with skeletal features suggestive of the otopalatodigital syndrome spectrum in boys. There appears to be a continuum among allelic disorders due to FLNA mutations.


Hydrogel-Sheathed Hipsc-Derived Heart Microtissue Enables Anchor-Free Contractile Force Measurement.
In vitro reconstruction of highly mature engineered heart tissues (EHTs) is attempted for the selection of cardiotoxic drugs suitable for individual patients before administration. Mechanical contractile force generated in the EHTs is known to be a critical indicator for evaluating the EHT response. However, measuring contractile force requires anchoring the EHT in a tailored force-sensing cell culture chamber, causing technical difficulties in the stable evaluation of contractile force in long-term culture. This paper proposes a hydrogel-sheathed human induced pluripotent stem cell (hiPSC)-derived heart microtissue (H3 M) that can provide an anchor-free contractile force measurement platform in commonly used multi-well plates. The contractile force associated with tissue formation and drug response is calculated by motion tracking and finite element analysis on the bending angle of the hydrogel sheath. From the experiment of the drug response, H3 M is an excellent drug screening platform with high sensitivity and early testing capability compared to conventionally anchored EHT. This unique platform would be useful and versatile for regenerative therapy and drug discovery research in EHT.


Fibroblast Growth Factor 21: A Fascinating Perspective on the Regulation of Muscle Metabolism.
Fibroblast growth factor 21 (FGF21) plays a vital role in normal eukaryotic organism development and homeostatic metabolism under the influence of internal and external factors such as endogenous hormone changes and exogenous stimuli. Over the last few decades, comprehensive studies have revealed the key role of FGF21 in regulating many fundamental metabolic pathways, including the muscle stress response, insulin signaling transmission, and muscle development. By coordinating these metabolic pathways, FGF21 is thought to contribute to acclimating to a stressful environment and the subsequent recovery of cell and tissue homeostasis. With the emphasis on FGF21, we extensively reviewed the research findings on the production and regulation of FGF21 and its role in muscle metabolism. We also emphasize how the FGF21 metabolic networks mediate mitochondrial dysfunction, glycogen consumption, and myogenic development and investigate prospective directions for the functional exploitation of FGF21 and its downstream effectors, such as the mammalian target of rapamycin (mTOR).


Chromosomal Arrangement of Phosphorelay Genes Couples Sporulation and DNA Replication.
Genes encoding proteins in a common regulatory network are frequently located close to one another on the chromosome to facilitate co-regulation or couple gene expression to growth rate. Contrasting with these observations, here, we demonstrate a functional role for the arrangement of Bacillus subtilis sporulation network genes on opposite sides of the chromosome. We show that the arrangement of two sporulation network genes, one located close to the origin and the other close to the terminus, leads to a transient gene dosage imbalance during chromosome replication. This imbalance is detected by the sporulation network to produce cell-cycle coordinated pulses of the sporulation master regulator Spo0A∼P. This pulsed response allows cells to decide between sporulation and continued vegetative growth during each cell cycle spent in starvation. The simplicity of this coordination mechanism suggests that it may be widely applicable in a variety of gene regulatory and stress-response settings. VIDEO ABSTRACT.


FCS study of the structural stability of lysozyme in the presence of morpholinium salts.
Ability of the ionic liquids to alter the structural stability of proteins in aqueous solution is a topic of considerable interest in modern bioscientific research because of possible applications of these substances in protein purification and as refolding agents. A few early studies involving the imidazolium ionic liquids have demonstrated their role as both denaturants and refolding agents. As the influence of an ionic liquid on a given protein depends on the identity of both species, it is necessary to extend the studies to a wider number of ionic liquids and proteins to obtain insight into the mechanism of interaction between the two and to arrive at a comprehensive picture. It is in this context that we have studied the effect of two morpholinium salts, [Mor1,2][Br] and [Mor1,4][Br], differing in the alkyl chain length of cation, on chicken egg white lysozyme in its native and chemically denatured states employing primarily the fluorescence correlation spectroscopy (FCS) technique. Fluorescence signal of Alexa488-labeled lysozyme (A488-Lysz) has been used to determine the changes in hydrodynamic radius of protein in the presence of additives. The results reveal a conformational dynamics of lysozyme with a time constant of 56 ± 10 μs in its native state. It is observed, when in its native state, both the morpholinium salts induce structural changes of lysozyme. However, when in its unfolded state, [Mor1,4][Br] at low concentration compacts the protein, but at higher concentration, it stabilizes the unfolded state, unlike [Mor1,2][Br], which compacts lysozyme at both low and high concentrations. A comparison of the effect of these salts and arginine, a protein stabilizer, on lysozyme indicates that [Mor1,2][Br] is a superior compacting agent for the unfolded state of the protein compared to arginine.


Positive psychological functioning in breast cancer: An integrative review.
This integrative review aimed to analyze the research into positive psychological functioning after breast cancer, and to integrate the most relevant findings relating to sociodemographic, medical and psychosocial factors. Relevant outcomes were identified from electronic databases (Medline, PsycINFO, Web of Science, Scopus, Cochrane, CINAHL, and Wiley Online Library) up to July 2015. A Google search was performed to identify unindexed literature. Dissertations and theses were searched on Proquest Dissertations and Theses, DIALNET and TDX. Selection criteria included empirical studies assessing relationships between breast cancer and positive functioning, without restrictions on type of participants. In total, 134 studies met the inclusion criteria. The sociodemographic, medical, and psychosocial characteristics associated with well-being, posttraumatic growth, finding benefit and meaning were being young, undergoing chemotherapy, and having social support. The last two of these characteristics were time-oriented. The culture of the different samples and positive dispositional characteristics like optimism had an influence on the women's coping styles. Socioeconomic status and level of education were also associated with positive psychological functioning. The perceived impact of breast cancer on patient, as well as the perceived support from significant others can result in better functioning in women with breast cancer. The results highlight that oncology health professionals should take into account not only the individual and medical characteristics, but also the stage of the oncological process and the psychosocial environment of patients in order to promote their positive functioning.


Ignoring spatial heterogeneity in drivers of SARS-CoV-2 transmission in the US will impede sustained elimination.
To dissect the transmission dynamics of SARS-CoV-2 in the United States, we integrate parallel streams of high-resolution data on contact, mobility, seasonality, vaccination and seroprevalence within a metapopulation network. We find the COVID-19 pandemic in the US is characterized by a geographically localized mosaic of transmission along an urban-rural gradient, with many outbreaks sustained by between-county transmission. We detect a dynamic tension between the spatial scale of public health interventions and population susceptibility as pre-pandemic contact is resumed. Further, we identify regions rendered particularly at risk from invasion by variants of concern due to spatial connectivity. These findings emphasize the public health importance of accounting for the hierarchy of spatial scales in transmission and the heterogeneous impacts of mobility on the landscape of contagion risk.


Relief from nitrogen starvation entails quick unexpected down-regulation of glycolytic/lipid metabolism genes in enological Saccharomyces cerevisiae.
Nitrogen composition of the grape must has an impact on yeast growth and fermentation kinetics as well as on the organoleptic properties of the final product. In some technological processes, such as white wine/rosé winemaking, the yeast-assimilable nitrogen content is sometimes insufficient to cover yeast requirements, which can lead to slow or sluggish fermentations. Growth is nevertheless quickly restored upon relief from nutrient starvation, e.g. through the addition of ammonium nitrogen, allowing fermentation completion. The aim of this study was to determine how nitrogen repletion affected the transcriptional response of a Saccharomyces cerevisiae wine yeast strain, in particular within the first hour after nitrogen addition. We found almost 4800 genes induced or repressed, sometimes within minutes after nutrient changes. Some of these responses to nitrogen depended on the TOR pathway, which controls positively ribosomal protein genes, amino acid and purine biosynthesis or amino acid permease genes and negatively stress-response genes, and genes related to the retrograde response (RTG) specific to the tricarboxylic acid (TCA) cycle and nitrogen catabolite repression (NCR). Some unexpected transcriptional responses concerned all the glycolytic genes, carbohydrate metabolism and TCA cycle-related genes that were down-regulated, as well as genes from the lipid metabolism.


Xeroderma pigmentosum-Cockayne syndrome complex.
Xeroderma pigmentosum-Cockayne syndrome complex is a very rare multisystem degenerative disorder (Orpha: 220295; OMIM: 278730, 278760, 278780, 610651). Published information on XP-CS is mostly scattered throughout the literature. We compiled statistics related to symptom prevalence in XP-CS and have written a clinical description of the syndrome. We also drew on clinical practices used in XP and in Cockayne syndrome without XP to aid management of XP-CS.Extensive searches of the literature identified 43 XP-CS patients. The diagnosis had been confirmed with molecular or biochemical methods in 42 of them. Clinical features of each patient were summarized in spreadsheets and summary statistics were generated from this data. XP patients are classified into complementation groups according to the gene that is mutated. There are four groups in XP-CS, and classification was available for 42 patients. Twenty-one were in the XP-G complementation group, 13 in XP-D, 5 in XP-B, and 3 in XP-F. Overall, the clinical features of XP-CS are very similar to those of CS without XP, with the exception of skin cancers in XP-CS. However, one intriguing finding was that cancer incidence was lower in XP-CS compared to XP alone or XP-neurological disorder. The cancer rate in XP-CS was higher than in CS without XP, an unsurprising finding. There is preliminary evidence for the existence of severity groups in XP-CS, as is the case in CS.Although health problems in XP-CS vary both in severity and in when they the first occur, there was overall homogeneity between all complementation groups and putative severity groups. Severely affected patients met fewer milestones and died at younger ages compared to more mildly affected patients.


Asymmetric Total Synthesis of Mycobacterial Diacyl Trehaloses Demonstrates a Role for Lipid Structure in Immunogenicity.
The first asymmetric total synthesis of three structures proposed for mycobacterial diacyl trehaloses, DAT1, DAT2, and DAT3 is reported. The presence of two of these glycolipids, DAT1 and DAT3, within different strains of pathogenic M. tuberculosis was confirmed, and it was shown that their abundance varies significantly. In mass spectrometry, synthetic DAT2 possessed almost identical fragmentation patterns to presumptive DAT2 from Mycobacterium tuberculosis H37Rv, but did not coelute by HPLC, raising questions as the precise relationship of the synthetic and natural materials. The synthetic DATs were examined as agonists for signaling by the C-type lectin, Mincle. The small differences in the chemical structure of the lipidic parts of DAT1, DAT2, and DAT3 led to drastic differences of Mincle binding and activation, with DAT3 showing similar potency as the known Mincle agonist trehalose dimycolate (TDM). In the future, DAT3 could serve as basis for the design of vaccine adjuvants with simplified chemical structure.


Deterministic SIR (Susceptible-Infected-Removed) models applied to varicella outbreaks.
Deterministic SIR models were applied to simulate Susceptible-Infected-Removed and to estimate the threshold condition for varicella outbreaks in children, reported in Medellín, Colombia. The expected numbers of susceptible, infected and removed individuals were compared with observed cases from notification of varicella outbreaks to the local Board of Health and from survey data. The threshold condition was estimated by the basic reproductive ratio and by the relative removal rate, through which measures for preventing and curtailing the outbreaks were identified. The model demonstrated a reasonable fit to the observations, except in two of the six outbreaks which probably reflected under-registration of cases. In order to have prevented these outbreaks, between 4.4% and 52.9% of the susceptible population should have been vaccinated assuming an 85% vaccine effectiveness. Similarly, isolation of affected children should have been increased to between 4.3% and 44.8% per week.


Comparing quantile residual life functions by confidence bands.
In this article we present a nonparametric method for constructing confidence bands for the difference of two quantile residual life (qrl) functions. These bands provide evidence for two random variables ordering with respect to the qrl order. The comparison of qrl functions is of importance, specially in the treatment of cancer when there exists a possibility of benefiting from a new secondary therapy. A qrl function is the quantile of the remaining life of a surviving subject, as it varies with time. We show the applicability of this approach in Medicine and Ecology. A simulation study has been carried out to evaluate and illustrate the performance and the consistency of this new methodology.


Evaluation of PC-ISO for customized, 3D Printed, gynecologic 192-Ir HDR brachytherapy applicators.
The purpose of this study was to evaluate the radiation attenuation properties of PC-ISO, a commercially available, biocompatible, sterilizable 3D printing material, and its suitability for customized, single-use gynecologic (GYN) brachytherapy applicators that have the potential for accurate guiding of seeds through linear and curved internal channels. A custom radiochromic film dosimetry apparatus was 3D-printed in PC-ISO with a single catheter channel and a slit to hold a film segment. The apparatus was designed specifically to test geometry pertinent for use of this material in a clinical setting. A brachytherapy dose plan was computed to deliver a cylindrical dose distribution to the film. The dose plan used an 192Ir source and was normalized to 1500 cGy at 1 cm from the channel. The material was evaluated by comparing the film exposure to an identical test done in water. The Hounsfield unit (HU) distributions were computed from a CT scan of the apparatus and compared to the HU distribution of water and the HU distribution of a commercial GYN cylinder applicator. The dose depth curve of PC-ISO as measured by the radiochromic film was within 1% of water between 1 cm and 6 cm from the channel. The mean HU was -10 for PC-ISO and -1 for water. As expected, the honeycombed structure of the PC-ISO 3D printing process created a moderate spread of HU values, but the mean was comparable to water. PC-ISO is sufficiently water-equivalent to be compatible with our HDR brachytherapy planning system and clinical workflow and, therefore, it is suitable for creating custom GYN brachytherapy applicators. Our current clinical practice includes the use of custom GYN applicators made of commercially available PC-ISO when doing so can improve the patient's treatment.


The radio-opacity of surgical and radiological devices used in vivo: a test method for markers in surgical gauze.
Most radio-opaque markers used in surgical gauze contain barium sulphate, and a minimum percentage content of this is required to ensure an acceptable level of radio-opacity. When this minimum percentage is specified by Standards authorities, a suitable test method must be available to determine compliance. A new radiographic test method is presented which can be used to determine the barium sulphate content or equivalence to within 2% of compliance requirements. The method consists of immersing the marker in a solution of barium sulphate which is then radiographed. Compliance can be evaluated by inspection of the radiograph. The method was found to be virtually independent of x-ray energy, film-screen combination and thickness or shape of the test object. The method is also an appropriate test for any other material or object, such as vascular catheters used in radiology or small toys which pose an ingestion hazard.


Similarities between plasmids of the P-incompatibility group derived from different bacterial genera.
The properties of four P-group plasmids, R26, R527, R751 and R906, which differ in resistance phenotype or in the bacterial species in which they were first detected, have been compared with the prototype of this group, RPI. Two of the plasmids, R26 and R527, are new isolates which have been assigned to the P group because of their incompatibility with R751. The properties studied include response to aeruginocin and to male and female sex-specific phages, interaction with prophage B3 and fertility inhibition by plasmid R38. Strains harbouring these plasmids behaved similarly in all tests except those involving aeruginocin. This suggests that the locus for aeruginocin-insensitivity is one of the R-determinants whereas the genes controlling the remaining characteristics are closely linked to the transfer factor. These plasmids may therefore have a common ancestor and their differences in resistance phenotype may simply reflect recombination events which they have undergone subsequently. Their similarity is also seen in transduction experiments, since determinants from two of these plasmids can be 'rescued' by the P-group plasmid R18 if this is already present in the recipient cell and the host recombination system is functional.


The etiologies and management of spinal cord compression in childhood cancers: Are we aware of the emergency of cord compression?
The spinal cord compression causes irreversible long-term permanent neurological sequelae. This study aims to increase awareness of childhood cancers that cause cord compression by comparing histopathological diagnosis, treatments, and survival rates to the literature. Seventy-three patients (38 male, 35 female) with spinal cord compression, among 1085 patients diagnosed with solid tumors at Gazi University Department of Pediatric Oncology between 1991 and 2021 were retrospectively evaluated. The mean time between the onset of complaints and diagnosis was 27.5± 24.9 (2-150) days. The first three most common tumors that caused cord compression; were central nervous system tumors in 22 (30%), neuroblastoma in 17 (23%), and malignant germ cell tumors in 8 (10%) cases. Of the patients, 46 (63%) had compression due to extradural masses, and 27 (37%) patients had an intradural compression. The most common symptoms were pain in 60 (82%), weakness in 57 (78%), and pins and needles in 28 (38%) patients, respectively. The clinical physical neurological examination findings were motor deficit in 62 (84%), and deep tendon reflex changes in 54 patients (73.9%). Compression findings were detected in 58 (79.5%) patients at diagnosis, and in 15 (20.5%) of them during follow-up. The most common level of compression was seen in the thoracolumbar region in 19 (26%) cases. In 65 (89%) patients with cord compression, corticosteroids were given as anti-edema treatment. Surgical excision was performed in 39 (53%) patients. Spinal radiotherapy was given to 35 patients (48%) with radiosensitive tumors. Chemotherapy protocols were started in 52 (71.2%) cases according to their diagnoses. Complete neurological recovery was achieved in 33 (45%) patients. The 5-year overall survival rates for solid tumors with extradural compression and intradural compression were 62% and 22%, respectively (p=0.002). Neurological sequela-free recovery is possible with early diagnosis and urgent treatment. Spinal compression must be detected by detailed systemic and neurological examination and imaging methods. Patients should be rapidly transferred to pediatric oncology units after starting anti-edema treatment.


Radionuclide bone scintigraphy in sports injuries.
Bone scintigraphy is one of the mainstays of molecular imaging. It has retained its relevance in the imaging of acute and chronic trauma and sporting injuries in particular. The basic reasons for its longevity are the high lesional conspicuity and technological changes in gamma camera design. The implementation of hybrid imaging devices with computed tomography scanners colocated with the gamma camera has revolutionized the technique by allowing a host of improvements in spatial resolution and anatomical registration. Both bone and soft-tissue lesions can be visualized and identified with greater and more convincing accuracy. The additional benefit of detecting injury before anatomical changes in high-level athletes has cost and performance advantages over other imaging modalities. The applications of the new imaging techniques will be illustrated in the setting of bone and soft-tissue trauma arising from sporting injuries.


Association between Lifestyle Changes and at-Home Hours during and after the State of Emergency Due to the COVID-19 Pandemic in Japan.
Lifestyle changes during the coronavirus disease (COVID-19) lockdown have been previously examined, but there is limited understanding about changes after such restrictions were lifted. This study examines changes in lifestyle habits and body weight among the Japanese population with regard to the length of at-home hours both during (April to May) and after (September) the nationwide stay-at-home request compared to those before the COVID-19 pandemic (January 2020). An online survey was conducted in September 2020 involving 10,000 Japanese survey monitors, selected according to population distribution. During the stay-at-home request, 34% participants extended their at-home hours. More respondents in the group with extended at-home hours experienced an increase or decrease in total physical activity, snacking, food intake, alcohol drinking, and body weight than those in the group with nonextended at-home hours. Some of these changes had a trend according to age. The prevalence of most of these changes decreased when at-home hours returned to normal after the stay-at-home request period; however, increased alcohol consumption and increased or decreased body weight persisted. Our findings suggest that close monitoring for further health outcomes and age-appropriate measures to encourage favorable health behaviors is needed.


[Features of genome expression of phage transposon D3112 of Pseudomonas aeruginosa in Escherichia coli bacteria: dependence of bacterial phenotype on copy number of D3112 genome].
Escherichia coli (RP4 :: D3112) bacteria manifest Tcs phenotype (thirty centigrade sensitivity), i.e. the cells do not divide and form colonies under conditions of lowered temperature (30 degrees C and lower), while cells grow normally at 42 degrees C. In this work it is demonstrated that replication-transposition of D3112 and the Tcs phenotype depend on no recA system of E.coli. Following events lead to the loss of the Tcs phenotype (in E.coli (RP4 :: D3112) cells survived after growing at 30 degrees C): occurrence of mutations in bacterial, phage and plasmid genomes, elimination of DNA of hybrid plasmid or RP4 DNA (a portion of DNA) as well as integration of the hybrid plasmid into bacterial chromosome. In the latter case, the E.coli (D3112) cells acquired the properties shared by the initial bacteria and those with the Tcs phenotype. Such clones are designated tcl (thirty centigrade low sensitivity), they are able to form colonies at 30 degrees C but their growth is more slow, they maintain instability at lowered temperature and continue to produce D3112 phage. The tcl clones in which replication-transposition of D3112 DNA in less effective than in the tcs clones are a suitable object for the study of genetic rearrangements caused by D3112 phage transposon. It is shown that either complete RP4 genome or its portion are comprised between direct repeats of D3112 and are built into various chromosomal sites, i.e. cointegrates are being formed. Two types of deletions are revealed: eliminating sites of RP4 plasmid adjacent to the left end of D3112 genome as well as deletions of the D3112 genome. It is demonstrated that alteration in the growth nature of E.coli, carrying D3112 DNA, at 30 degrees C depends on the copy number of D3112 per bacterial cell.


Pure Laparoscopic Anatomic Resection of Liver Segment 4 with Middle Hepatic Vein Involvement Using Indocyanine Green Fluorescence Staining.
Laparoscopic anatomic resection of liver segment 4 is a technically challenging operation, which is rarely reported owing to the difficulty of defining the demarcation of a hepatic segment 4 on a monitor.1 The portal territory staining method is technically feasible to identify tumors and segment boundaries during hepatectomy.2 Herein, we describe the laparoscopic hepatectomy of segment 4 using the fluorescent-positive staining method. A 72-year-old man recurred colorectal liver metastases after colectomy, positron emission tomography (PET)/computed tomography (CT) showed metastases located in segment 4 with involvement of the middle hepatic vein (MHV) and caudate lobe; no other organ metastasis or recurrence occurred. We performed an anatomical hepatectomy 4 with MHV and parenchymal resection of segment 1 (H1'/4-MHV).3 The key point of the procedure was dividing and clamping Glisson's branches for segment 2 and segment 3 using the hepatic round ligament approach; the G2 and G3 were dissected along the right side of round ligament via the extrahepatic Glissonian approach, then the left hepatic artery (LHA) was divided and injected with ICG in the left portal vein (LPV). Finally, transection was performed along the fluorescent stain location line and ischemic demarcation line. The operation time was 263 min; the Pringle lasted 110 min, and the estimated blood loss was 400 g. The patient was discharged on postoperative day 5 without complications. Sigmoid carcinoma and R0 margin were confirmed by histopathology. Laparoscopic anatomic hepatectomy 4 with middle hepatic vein invasion using indocyanine green (ICG) fluorescence staining is a feasible and effective technique.


Emerging applications of metabolomics in studying chemopreventive phytochemicals.
Phytochemicals from diet and herbal medicines are under intensive investigation for their potential use as chemopreventive agents to block and suppress carcinogenesis. Chemical diversity of phytochemicals, together with complex metabolic interactions between phytochemicals and biological system, can overwhelm the capacity of traditional analytical platforms, and thus pose major challenges in studying chemopreventive phytochemicals. Recent progresses in metabolomics have transformed it to become a robust systems biology tool, suitable for examining both chemical and biochemical events that contribute to the cancer prevention activities of plant preparations or their bioactive components. This review aims to discuss the technical platform of metabolomics and its existing and potential applications in chemoprevention research, including identifying bioactive phytochemicals in plant extracts, monitoring phytochemical exposure in humans, elucidating biotransformation pathways of phytochemicals, and characterizing the effects of phytochemicals on endogenous metabolism and cancer metabolism.


Synthesis, biological evaluation, and docking studies of some 5-chloro-2(3H)-benzoxazolone Mannich bases derivatives as cholinesterase inhibitors.
A series of N-substituted-5-chloro-2(3H)-benzoxazolone derivatives were synthesized and evaluated for their acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) inhibitory, and antioxidant activities. The structures of the title compounds were confirmed by spectral and elemental analyses. The cholinesterase (ChE) inhibitory activity studies were carried out using Ellman's colorimetric method. The free radical scavenging activity was also determined by in vitro ABTS (2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)) assay. The biological activity results revealed that all of the title compounds displayed higher AChE inhibitory activity than the reference compound, rivastigmine, and were selective for AChE. Among the tested compounds, compound 7 exhibited the highest inhibition against AChE (IC50 = 7.53 ± 0.17 μM), while compound 11 was found to be the most active compound against BuChE (IC50 = 17.50 ± 0.29 μM). The molecular docking study of compound 7 showed that this compound can interact with the catalytic active site (CAS) of AChE and also has potential metal chelating ability and a proper log P value. On the other hand, compound 2 bearing a methyl substituent at the ortho position on the phenyl ring showed better radical scavenging activity (IC50 = 1.04 ± 0.04 mM) than Trolox (IC50 = 1.50 ± 0.05 mM).


The role of closed-packed positions in the pathogenesis of temporomandibular joint internal derangements.
A hypothesis is presented as to how certain occlusal relationships and habitual patterns of jaw use may predispose an individual to TMJ internal derangements. Knowledge of diarthrodial joint biomechanics, in general, and the appreciation of close-packed positions of the TMJ, in particular, would not only provide the clinician with a greater discriminatory sense when diagnosing intracapsular problems but also help him to make better decisions and judgement about therapy. Popular treatment regimens, in particular those based upon the age-old Centric Relation concept, will be re-appraised.


Isolation and sequence analysis of the Chlamydia pneumoniae GroE operon.
Chlamydia pneumoniae has emerged as an important human respiratory pathogen. From a lambda gt11 gene bank constructed from C. pneumoniae isolate AR-39 DNA, an immunoreactive plaque containing a 3.0-kb insert was purified. In immunoblots, a 60-kDa protein was recognized by anti-C. pneumoniae rabbit immune serum. The recombinant protein was reactive with a Chlamydia genus-specific monoclonal antibody recognizing a 60-kDa protein found in the Sarkosyl-soluble fraction and with rabbit immune serum prepared against the Chlamydia trachomatis 60-kDa GroEL homolog associated with the delayed-type hypersensitivity response. DNA sequence analysis confirmed that the C. pneumoniae gene product is an analog of the C. trachomatis delayed-type hypersensitivity antigen and the Escherichia coli GroEL heat shock protein.


Endothelialization of a new Dacron graft in an experimental model: light microscopy, electron microscopy and immunocytochemistry.
Two types of synthetic vascular grafts, Dacron Triaxial and Dacron Gelseal Triaxial, were implanted into both the common carotids of sheep. The animals were sacrificed 1, 2, 8, and 16 weeks after surgery. Multiple specimens, obtained from grafts and anastomoses, were studied by light microscopy, transmission and scanning electron microscopy. A parallel immunocytochemical analysis was performed on some specimens. Dacron Triaxial grafts failed to develop a complete neointimal coverage. Myofibroblasts and fibroblasts were the dominant cells in such synthetic graft. Moreover, focal areas of stripping, platelet deposition, and thrombosis were observed at 8 and 16 weeks. In contrast, a stable endothelial coverage developed on the Gelseal Triaxial grafts after 16 weeks.


[Development cycle of the yeastlike fungus, Endomycopsis fibuligera, and its accumulation of glucoamylase activity].
The growth cycle and the accumulation of glucoamylase by the yeast-like fungus Endomycopsis fibuligera 20-9 were studied in the conditions of submerged cultivation. The enzyme was accumulated by the end of the logarithmic phase and particularly at the stationary phase of the cultural growth. A correlation has been established between the accumulation of the glucoamylase in the cultural broth and the production of blastospores. Possible systems involved in the control of the enzyme accumulation are discussed.


A pattern-based approach to the interpretation of skeletal muscle biopsies.
The interpretation of muscle biopsies is complex and provides the most useful information when integrated with the clinical presentation of the patient. These biopsies are performed for workup of a wide range of diseases including dystrophies, metabolic diseases, and inflammatory processes. Recent insights have led to changes in the classification of inflammatory myopathies and have changed the role that muscle biopsies have in the workup of inherited diseases. These changes will be reviewed. This review follows a morphology-driven approach by discussing diseases of skeletal muscle based on a few basic patterns that include cases with (1) active myopathic damage and inflammation, (2) active myopathic damage without associated inflammation, (3) chronic myopathic changes, (4) myopathies with distinctive inclusions or vacuoles, (5) biopsies mainly showing atrophic changes, and (6) biopsies that appear normal on routine preparations. Each of these categories goes along with certain diagnostic considerations and pitfalls. Individual biopsy features are only rarely pathognomonic. Establishing a firm diagnosis therefore typically requires integration of all of the biopsy findings and relevant clinical information. With this approach, a muscle biopsy can often provide helpful information in the diagnostic workup of patients presenting with neuromuscular problems.


MAVS Promotes Inflammasome Activation by Targeting ASC for K63-Linked Ubiquitination via the E3 Ligase TRAF3.
Stringent control of inflammasome signaling pathway is important for maintaining immunological balance, yet the molecular mechanisms responsible for its tight regulation are still poorly understood. In this study, we found that the signaling pathway dependent on mitochondrial antiviral signaling protein (MAVS) was required for the optimal activation of apoptosis-associated specklike protein (ASC)-dependent inflammasome. In particular, TNFR-associated factor 3 was found to be a direct E3 ligase for ASC. Ubiquitination of ASC at Lys(174) was critical for speck formation and inflammasome activation. Deficiency in MAVS or TNFR-associated factor 3 impaired ASC ubiquitination and cytosolic aggregates formation, resulting in reduced inflammasome response upon RNA virus infection. This study has identified a previously unrecognized role of MAVS in the regulation of inflammasome signaling and provided molecular insight into the mechanisms by which ubiquitination of ASC controls inflammasome activity through the formation of ASC specks.


Environmental risk factors and Parkinson's disease: a case-control study in Taiwan.
To explore environmental risk factors for Parkinson's disease (PD) in Taiwan, we investigated 120 patients with PD and 240 hospital control subjects matched with patients on age (+/-2 years) and sex. Based on a structured open-ended questionnaire, we carried out standardized interviews to obtain history of exposure to environmental factors, including place of residence, source of drinking water, and environmental and occupational exposures to various agricultural chemicals. In the univariate analysis, the history of living in a rural environment, farming, use of herbicides/pesticides, and use of paraquat were associated with an increased PD risk in a dose-response relationship. After adjustment for multiple risk factors through conditional logistic regression, the biological gradient between PD and previous uses of herbicides/pesticides and paraquat remained significant. The PD risk was greater among subjects who had used paraquat and other herbicides/pesticides than those who had used herbicides/pesticides other than paraquat. There were no significant differences in occupational exposures to chemicals, heavy metals, and minerals between PD patients and matched control subjects. The duration of drinking well water and alcohol consumption was not significantly associated with PD. There was an inverse relationship between cigarette smoking and PD. Environmental factors, especially exposures to paraquat and herbicides/pesticides, may play important roles in the development of PD in Taiwan.


Accumulation of rhodopsin in late endosomes triggers photoreceptor cell degeneration.
Progressive retinal degeneration is the underlying feature of many human retinal dystrophies. Previous work using Drosophila as a model system and analysis of specific mutations in human rhodopsin have uncovered a connection between rhodopsin endocytosis and retinal degeneration. In these mutants, rhodopsin and its regulatory protein arrestin form stable complexes, and endocytosis of these complexes causes photoreceptor cell death. In this study we show that the internalized rhodopsin is not degraded in the lysosome but instead accumulates in the late endosomes. Using mutants that are defective in late endosome to lysosome trafficking, we were able to show that rhodopsin accumulates in endosomal compartments in these mutants and leads to light-dependent retinal degeneration. Moreover, we also show that in dying photoreceptors the internalized rhodopsin is not degraded but instead shows characteristics of insoluble proteins. Together these data implicate buildup of rhodopsin in the late endosomal system as a novel trigger of death of photoreceptor neurons.


Evaluation of Newly Integrated Bivalirudin Titration Protocol in Patients With Mechanical Circulatory Support.
Patients with cardiogenic shock or end-stage heart failure can be maintained on mechanical circulatory support (MCS) devices. Once a patient undergoes placement of a device, obtaining and maintaining therapeutic anticoagulation is vital. Guidelines recommend the use of institutional protocols to assist in dosing and titration of anticoagulants. The purpose of this study was to characterize the use of bivalirudin before and after the implementation of a standardized titration protocol in patients with MCS. A retrospective review of patients who received bivalirudin for MCS (VA ECMO [veno-arterial extracorporeal membrane oxygenation], Impella, or LVAD [left ventricular assist device]) before and after the implementation of the titration protocol into the electronic health record (EHR) was conducted. The primary outcome was to compare the proportion of therapeutic activated partial thromboplastin time (aPTT). Secondary outcomes included number of subtherapeutic and supratherapeutic aPTTs, incidence of bleeding and clotting events, bivalirudin titrations per day, and percentage of patients with therapeutic aPTT level. A total of 100 patients were included (precohort = 67; postcohort = 33). The proportion of therapeutic aPTTs was significantly higher in the postcohort than that in the precohort (62% vs 48%; P < 0.001). The postcohort had 0% of patients failing to achieve therapeutic aPTT levels. The number of titrations per day was significantly lower in the postcohort, with 1.20 titrations per day versus 1.93 in the precohort (P < 0.001). Implementation of the bivalirudin titration nomograms within the EHR significantly increased the number of therapeutic aPTTs, reduced the number of patients who never achieved a therapeutic aPTT, and reduced the required number of titrations per day.


Gathering and learning from relevant clinical data: a new framework.
Given the rising costs of health care in today's economic environment, the need for effective, value-driven care has never been more pressing. While the U.S. health care system strives continually to improve patient outcomes, it struggles with the inadequacies due to variation in care and the inefficiencies of unnecessary resource utilization. The tools traditionally used to study care, from retrospective studies to randomized controlled trials, may be inadequate to address the complicated, interdependent questions related to defining effective care. To overcome the deficiencies of these traditional tools and better optimize our health care system, a new kind of methodology is required--one that integrates the functionality of previously existing tools in a novel way. Standardized Clinical Assessment and Management Plans (SCAMPs) were designed to accomplish this goal. A SCAMP is a care pathway, designed by clinicians, to guide medical decision making around a particular disorder. SCAMPs are unique in that they invite knowledge-based diversions from their recommendations and are accompanied by data collection and continuous improvement processes. Through these mechanisms, SCAMPs successfully reduce practice variation, optimize resource use, and create an integrated medical learning system which overcomes many of the inadequacies of traditional research tools. As such, the SCAMP paradigm may represent an important breakthrough in the effort to define and implement effective health care.