Nutritional sustainability of pet foods.
Sustainable practices meet the needs of the present without compromising the ability of future generations to meet their needs. Applying these concepts to food and feed production, nutritional sustainability is the ability of a food system to provide sufficient energy and essential nutrients required to maintain good health in a population without compromising the ability of future generations to meet their nutritional needs. Ecological, social, and economic aspects must be balanced to support the sustainability of the overall food system. The nutritional sustainability of a food system can be influenced by several factors, including the ingredient selection, nutrient composition, digestibility, and consumption rates of a diet. Carbon and water footprints vary greatly among plant- and animal-based ingredients, production strategy, and geographical location. Because the pet food industry is based largely on by-products and is tightly interlinked with livestock production and the human food system, however, it is quite unique with regard to sustainability. Often based on consumer demand rather than nutritional requirements, many commercial pet foods are formulated to provide nutrients in excess of current minimum recommendations, use ingredients that compete directly with the human food system, or are overconsumed by pets, resulting in food wastage and obesity. Pet food professionals have the opportunity to address these challenges and influence the sustainability of pet ownership through product design, manufacturing processes, public education, and policy change. A coordinated effort across the industry that includes ingredient buyers, formulators, and nutritionists may result in a more sustainable pet food system.


Metabolic benefits of annatto-extracted tocotrienol on glucose homeostasis, inflammation, and gut microbiome.
Emerging evidence suggests that the gut microbiome plays an important role in the pathophysiology of both obesity and type 2 diabetes mellitus. We previously reported that dietary annatto-extracted tocotrienol exerts beneficial effects by modulating inflammatory responses in mice fed a high-fat diet (HFD). The purpose of this study was to test the hypothesis that tocotrienol supplementation when combined with an HFD would result in an altered gut microbiota composition. For 14 weeks, forty-eight male C57BL/6J mice were assigned to 4 groups-low-fat diet, HFD, HFD supplemented with annatto-extracted tocotrienol at 800 mg/kg diet (AT), and HFD supplemented with metformin at 200 mg/kg diet. Glucose homeostasis was assessed by glucose and insulin tolerance tests, serum and pancreas insulin levels, and histological assessments of insulin and glucagon in pancreatic tissue. The concentrations of adipokines were measured in white adipose tissues. For the gut microbiome analysis, cecal content was collected, DNA was extracted, and 16S rRNA gene sequencing was performed. AT supplementation improved glucose homeostasis and lowered resistin, leptin, and interleukin-6 levels in white adipose tissue. Relative to the HFD group, AT-supplemented mice showed a decrease in the Firmicutes to Bacteroidetes ratio and had a lower abundance of Ruminococcus lactaris, Dorea longicatena, and Lachnospiraceae family. The relative abundance of Akkermansia muciniphila was increased in the AT group compared to the low-fat diet group. The association between the metabolic improvements and the identified bacterial taxa suggests a potential metabolic modulation caused by AT supplementation through the gut microbiota composition in mice fed an HFD.


Murine norovirus replicase augments RIG-I-like receptors-mediated antiviral interferon response.
Noroviruses are the main causative agents for acute viral gastroenteritis worldwide. RIG-I-like receptors (RLRs) triggered interferon (IFN) activation is essential for host defense against viral infections. In turn, viruses have developed sophisticated strategies to counteract host antiviral response. This study aims to investigate how murine norovirus (MNV) replicase interacts with RLRs-mediated antiviral IFN response. Counterintuitively, we found that the MNV replicase NS7 enhances the activation of poly (I:C)-induced IFN response and the transcription of downstream interferon-stimulated genes (ISGs). Interestingly, NS7 protein augments RIG-I and MDA5-triggered antiviral IFN response, which conceivably involves direct interactions with the caspase activation and recruitment domains (CARDs) of RIG-I and MDA5. Consistently, RIG-I and MDA5 exert anti-MNV activity in human HEK293T cells with ectopic expression of viral receptor CD300lf. This effect requires the activation of JAK/STAT pathway, and is further enhanced by NS7 overexpression. These findings revealed an unconventional role of MNV NS7 as augmenting RLRs-mediated IFN response to inhibit viral replication.


A model-based analysis of impulsivity using a slot-machine gambling paradigm.
Impulsivity plays a key role in decision-making under uncertainty. It is a significant contributor to problem and pathological gambling (PG). Standard assessments of impulsivity by questionnaires, however, have various limitations, partly because impulsivity is a broad, multi-faceted concept. What remains unclear is which of these facets contribute to shaping gambling behavior. In the present study, we investigated impulsivity as expressed in a gambling setting by applying computational modeling to data from 47 healthy male volunteers who played a realistic, virtual slot-machine gambling task. Behaviorally, we found that impulsivity, as measured independently by the 11th revision of the Barratt Impulsiveness Scale (BIS-11), correlated significantly with an aggregate read-out of the following gambling responses: bet increases (BIs), machines switches (MS), casino switches (CS), and double-ups (DUs). Using model comparison, we compared a set of hierarchical Bayesian belief-updating models, i.e., the Hierarchical Gaussian Filter (HGF) and Rescorla-Wagner reinforcement learning (RL) models, with regard to how well they explained different aspects of the behavioral data. We then examined the construct validity of our winning models with multiple regression, relating subject-specific model parameter estimates to the individual BIS-11 total scores. In the most predictive model (a three-level HGF), the two free parameters encoded uncertainty-dependent mechanisms of belief updates and significantly explained BIS-11 variance across subjects. Furthermore, in this model, decision noise was a function of trial-wise uncertainty about winning probability. Collectively, our results provide a proof of concept that hierarchical Bayesian models can characterize the decision-making mechanisms linked to the impulsive traits of an individual. These novel indices of gambling mechanisms unmasked during actual play may be useful for online prevention measures for at-risk players and future assessments of PG.


Stability of cognitive performance in children with mild intellectual disability.
Longitudinal studies that have examined cognitive performance in children with intellectual disability more than twice over the course of their development are scarce. We assessed population and individual stability of cognitive performance in a clinical sample of children with borderline to mild non-syndromic intellectual disability. Thirty-six children (28 males, eight females; age range 3-19y) with borderline to mild intellectual disability (Full-scale IQ [FSIQ] 50-85) of unknown origin were examined in a retrospective clinical case series using linear mixed models including at least three assessments with standardized intelligence tests. Average cognitive performance remained remarkably stable over time (high population stability, drop of only 0.38 IQ points per year, standard error=0.39, p=0.325) whereas individual stability was at best moderate (intraclass correlation of 0.58), indicating that about 60% of the residual variation in FSIQ scores can be attributed to between-child variability. Neither sex nor socio-economic status had a statistically significant impact on FSIQ. Although intellectual disability during childhood is a relatively stable phenomenon, individual stability of IQ is only moderate, likely to be caused by test-to-test reliability (e.g. level of child's cooperation, motivation, and attention). Therefore, clinical decisions and predictions should not rely on single IQ assessments, but should also consider adaptive functioning and previous developmental history.


A micro RNA processing defect in rapidly progressing idiopathic pulmonary fibrosis.
Idiopathic pulmonary fibrosis exhibits differential progression from the time of diagnosis but the molecular basis for varying progression rates is poorly understood. The aim of the present study was to ascertain whether differential miRNA expression might provide one explanation for rapidly versus slowly progressing forms of IPF. miRNA and mRNA were isolated from surgical lung biopsies from IPF patients with a clinically documented rapid or slow course of disease over the first year after diagnosis. A quantitative PCR miRNA array containing 88 of the most abundant miRNA in the human genome was used to profile lung biopsies from 9 patients with rapidly progressing IPF, 6 patients with slowly progressing IPF, and 10 normal lung biopsies. Using this approach, 11 miRNA were significantly increased and 36 were significantly decreased in rapid biopsies compared with normal biopsies. Slowly progressive biopsies exhibited 4 significantly increased miRNA and 36 significantly decreased miRNA compared with normal lung. Among the miRNA present in IPF with validated mRNA targets were those with regulatory effects on epithelial-mesenchymal transition (EMT). Five miRNA (miR-302c, miR-423-5p, miR-210, miR-376c, and miR-185) were significantly increased in rapid compared with slow IPF lung biopsies. Additional analyses of rapid biopsies and fibroblasts grown from the same biopsies revealed that the expression of AGO1 and AGO2 (essential components of the miRNA processing RISC complex) were lower compared with either slow or normal lung biopsies and fibroblasts. These findings suggest that the development and/or clinical progression of IPF might be the consequence of aberrant miRNA processing.


Sense of humor among the elderly: findings using the German version of the SHS.
Empirical studies on humor among the elderly are lacking. Based on the model of sense of humor by Paul McGhee, different age groups are compared (n=979) in the present study. Data suggest that playfulness and sense of humor are stable across different age groups (in a cross-sectional design) but that elderly participants were highest in positive mood. However, they also indicated that they laughed less and less easily than younger participants. Among the elderly, those who frequently (more than once a week) meet with friends and colleagues yielded higher expressions in different aspects of the sense of humor than those who meet friends and colleagues less often. Generally, humor was positively associated with personal and national well-being. However, among those participants older than 60 years of age, national well-being (i.e., satisfaction with the government, safety, or economics in the country) was negatively related to humor. Results are discussed together with a general outlook on the use of humor interventions for increasing the well-being and quality of life of elderly people.


Adaptive control of laser modal properties.
An adaptive optical system for precise control of a laser beam's mode structure has been developed. The system uses a dynamic lens based on controlled optical path deformation in a dichroic optical element that is heated with an auxiliary laser. Our method is essentially aberration free, has high dynamic range, and can be implemented with high average power laser beams where other adaptive optics methods fail. A quantitative model agrees well with our experimental data and demonstrates the potential of our method as a mode-matching and beam-shaping element for future large-scale gravitational wave detectors.


ESAT-6 proteins: protective antigens and virulence factors?
The 6kDa early secreted antigenic target from Mycobacterium tuberculosis, ESAT-6, is the prototype of a novel family of small proteins of unknown function produced by Actinobacteria. Export of ESAT-6, a potent T-cell antigen, and related proteins requires a dedicated secretory apparatus that is encoded by a cluster of genes, several of which also code for proteins that are recognized strongly by T cells. ESAT-6 systems can thus be considered as immunogenicity islands and there is growing evidence that the corresponding genes are subject to selective pressure imposed by the immune system of the host. Recently, there has been major progress in understanding the biogenesis, secretion and antigenicity of ESAT-6 proteins and, at least in the case of ESAT-6 system 1, in unravelling their role in pathogenicity. Here, we discuss these findings and their implications for the development of new therapeutic and prophylactic interventions against tuberculosis.


Molecular Diagnosis of Diffuse Gliomas through Sequencing of Cell-Free Circulating Tumor DNA from Cerebrospinal Fluid.
Purpose: Diffuse gliomas are the most common primary tumor of the brain and include different subtypes with diverse prognosis. The genomic characterization of diffuse gliomas facilitates their molecular diagnosis. The anatomical localization of diffuse gliomas complicates access to tumor specimens for diagnosis, in some cases incurring high-risk surgical procedures and stereotactic biopsies. Recently, cell-free circulating tumor DNA (ctDNA) has been identified in the cerebrospinal fluid (CSF) of patients with brain malignancies.Experimental Design: We performed an analysis of IDH1, IDH2, TP53, TERT, ATRX, H3F3A, and HIST1H3B gene mutations in two tumor cohorts from The Cancer Genome Atlas (TCGA) including 648 diffuse gliomas. We also performed targeted exome sequencing and droplet digital PCR (ddPCR) analysis of these seven genes in 20 clinical tumor specimens and CSF from glioma patients and performed a histopathologic characterization of the tumors.Results: Analysis of the mutational status of the IDH1, IDH2, TP53, TERT, ATRX, H3F3A, and HIST1H3B genes allowed the classification of 79% of the 648 diffuse gliomas analyzed, into IDH-wild-type glioblastoma, IDH-mutant glioblastoma/diffuse astrocytoma and oligodendroglioma, each subtype exhibiting diverse median overall survival (1.1, 6.7, and 11.2 years, respectively). We developed a sequencing platform to simultaneously and rapidly genotype these seven genes in CSF ctDNA allowing the subclassification of diffuse gliomas.Conclusions: The genomic analysis of IDH1, IDH2, TP53, ATRX, TERT, H3F3A, and HIST1H3B gene mutations in CSF ctDNA facilitates the diagnosis of diffuse gliomas in a timely manner to support the surgical and clinical management of these patients. Clin Cancer Res; 24(12); 2812-9. ©2018 AACR.


Modern Management and Diagnosis of Hypertension in the United Kingdom: Home Care and Self-care.
The effective diagnosis and management of hypertension is one of the most important parts of cardiovascular prevention internationally and this is no different in the United Kingdom. Approximately 14% of the UK population currently receive treatment for hypertension. Recent UK guidelines from the National Institute of Health and Care Excellence have placed greater emphasis on the utilization of out-of-office measurement of blood pressure to more accurately diagnose hypertension. The aim of the present study was to provide a state-of-the-art review of the evidence for screening, diagnosing, and managing hypertension, as implemented in the United Kingdom, with an emphasis on the role of self-monitored and ambulatory blood pressure monitoring in routine clinical care. Consideration was given to the use of ambulatory and home monitoring to confirm a diagnosis of hypertension and the use of self-monitoring and self-management to monitor and guide treatment. The evidence for the use of self-monitoring in patients with hypertension was examined, both in isolation, and in combination with lifestyle and treatment interventions. There is a place for self-monitored blood pressure in specific underresearched populations such as the elderly, specialist conditions, ethnic groups, and during pregnancy and this is discussed here. The evidence supporting the use of out-of-office monitoring in all aspects of routine clinical care has increased substantially in recent years and is reflected in increased utilization by patients and clinicians alike. Several areas require further research but it is clear that out-of-office monitoring is here to stay and is fast becoming an important part of hypertension management in the United Kingdom.


Turning on/off satellite droplet ejection for flexible sample delivery on digital microfluidics.
Digital microfluidics has the potential to minimize and automate reactions in biochemical labs. However, the complexity of drop manipulation and sample preparation on-chip has limited its incorporation into daily workflow. In this paper, we report a novel method for flexible sample delivery on digital microfluidics in a wide volume range spanning four orders of magnitude from picoliters to nanoliters. The method is based on the phenomenon of satellite droplet ejection, triggered by a sudden change in the strength of the electric field across a drop on a hydrophobic dielectric surface. By precisely modulating the actuation signal with convenient external electric controls, satellite droplet ejection can be turned on to dispense samples or turned off to transport picking-up drops. A pico-dosing design is presented and validated in this work to demonstrate the direct and flexible on-chip sample delivery. This approach could pave the way for the acceptance of microfluidics as a common platform for daily reactions to realize lab-on-a-chip.


Direct oral anticoagulants: new drugs and new concepts.
Direct oral anticoagulants (DOACs) are approved for multiple thromboembolic disorders and provide advantages over existing agents. As with all anticoagulants, management protocols for the eventuality of bleeding are important. Randomized phase III studies generally show that DOACs have a similar risk of clinically relevant bleeding compared with standard anticoagulants, with reductions in major bleeding in some cases. This may be particularly important in patients with atrial fibrillation, for whom the rate of intracranial hemorrhage was approximately halved with DOACs compared with warfarin. Conversely, the risk of gastrointestinal bleeding may be increased. Specific patient characteristics, such as renal impairment, comedications, and particular aspects of each drug, including the proportion eliminated by the kidneys, must be taken into account when assessing the risk of bleeding. Although routine coagulation monitoring of DOACs is not required, it may be useful under some circumstances. Of the traditional clotting assays, a sensitive and calibrated prothrombin time may be useful for detecting the presence or absence of clinically relevant factor Xa inhibitor concentrations (rivaroxaban or apixaban), but specific anti-factor Xa assays can measure drug levels quantitatively. For dabigatran, the results of an activated partial thromboplastin time test may exclude a clinically relevant pharmacodynamic effect, but a calibrated dilute thrombin time assay can be used for quantification of drug levels. In the event of mild or moderate bleeding, normal hemostatic support measures are recommended. For life-threatening bleeding, use of nonspecific prohemostatic agents may be considered, although clinical evidence is scarce. Specific antidotes are in development.


Characterization of tibetan medicine zuota by multiple techniques.
Zuota is regarded as the king of Tibetan medicine. However, due to the confidentiality of this precious medicine, the scientific characterization of Zuota is very scarce, which limits the pharmacology and biosafety studies of Zuota. Herein, we collected four different Zuota samples from Tibet, Qinghai, Gansu, and Sichuan and characterized them by multiple techniques. Our results showed that Zuota was mainly an inorganic mixture of HgS, sulfur, and graphite. Morphologically, Zuota samples were composed of nanoparticles, which further aggregated into microsized particles. Chemically, the majorities of Zuota were S and Hg (in the forms of HgS and pure sulfur). All samples contained pure sulfur with orthorhombic crystalline. Zuota from Qinghai province had different HgS crystalline, namely, hexagonal crystalline. The others were all face-centered cubic crystalline. Carbon in Zuota NPs was in the form of graphite. The implication to future studies of Zuota was discussed.


Conflicting processes in the evolution of body size and development time.
Body size and development time of Manduca sexta are both determined by the same set of three developmental-physiological factors. These define a parameter space within which it is possible to analyse and explain how phenotypic change is associated with changes in the underlying factors. Body size and development time are determined by the identical set of underlying factors, so they are not independent, but because the mechanisms by which these factors produce each phenotype are different, the two phenotypes are only weakly correlated, and the correlation is context dependent. We use a mathematical model of this mechanism to explore the association between body size and development time and show that the correlation between these two life-history traits can be positive, zero or negative, depending entirely on where in parameter space a population is located, and on which of the underlying factors has a greater variation. The gradient within this parameter space predicts the unconstrained evolutionary trajectory under directional selection on each trait. Calculations of the gradients for body size and development time revealed that these are nearly orthogonal through much of the parameter space. Therefore, simultaneous directional selection on body size and development time can be neither synergistic nor antagonistic but leads to conflicting selection on the underlying developmental parameters.


Impaired hepatic release of triglycerides: a possible cause of acute alcoholic fatty liver in the Rhesus monkey.
1. The mechanism of development of acute alcoholic fatty liver in the Rhesus monkey was investigated by studying the effect of Triton WR-1339 on plasma triglycerides. 2. After ethanol infusion, the rise in plasma triglyceride produced by Triton was greatly reduced and there was an accumulation of triglyceride in the liver. 3. Thus findings suggest that acute alcoholic fatty liver in Rhesus monkeys is probably due to a defect in the release of triglyceride from the liver.


Pomalidomide mitigates neuronal loss, neuroinflammation, and behavioral impairments induced by traumatic brain injury in rat.
Traumatic brain injury (TBI) is a global health concern that typically causes emotional disturbances and cognitive dysfunction. Secondary pathologies following TBI may be associated with chronic neurodegenerative disorders and an enhanced likelihood of developing dementia-like disease in later life. There are currently no approved drugs for mitigating the acute or chronic effects of TBI. The effects of the drug pomalidomide (Pom), an FDA-approved immunomodulatory agent, were evaluated in a rat model of moderate to severe TBI induced by controlled cortical impact. Post-TBI intravenous administration of Pom (0.5 mg/kg at 5 or 7 h and 0.1 mg/kg at 5 h) was evaluated on functional and histological measures that included motor function, fine more coordination, somatosensory function, lesion volume, cortical neurodegeneration, neuronal apoptosis, and the induction of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6). Pom 0.5 mg/kg administration at 5 h, but not at 7 h post-TBI, significantly mitigated the TBI-induced injury volume and functional impairments, neurodegeneration, neuronal apoptosis, and cytokine mRNA and protein induction. To evaluate underlying mechanisms, the actions of Pom on neuronal survival, microglial activation, and the induction of TNF-α were assessed in mixed cortical cultures following a glutamate challenge. Pom dose-dependently ameliorated glutamate-mediated cytotoxic effects on cell viability and reduced microglial cell activation, significantly attenuating the induction of TNF-α. Post-injury treatment with a single Pom dose within 5 h significantly reduced functional impairments in a well-characterized animal model of TBI. Pom decreased the injury lesion volume, augmented neuronal survival, and provided anti-inflammatory properties. These findings strongly support the further evaluation and optimization of Pom for potential use in clinical TBI.


Alveolar ridge and maxillary sinus augmentation using rhBMP-2: a systematic review.
The aim of this systematic review was to evaluate clinical and safety data for recombinant human bone morphogenetic protein-2 (rhBMP-2) in an absorbable collagen sponge (ACS) carrier when used for alveolar ridge/maxillary sinus augmentation in humans. Clinical studies/case series published 1980 through June 2012 using rhBMP-2/ACS were searched. Studies meeting the following criteria were considered eligible for inclusion: >10 subjects at baseline and maxillary sinus or alveolar ridge augmentation not concomitant with implant placement. Seven of 69 publications were eligible for review. rhBMP-2/ACS yielded clinically meaningful bone formation for maxillary sinus augmentation that would allow placement of regular dental implants without consistent differences between rhBMP-2 concentrations. Nevertheless, the statistical analysis showed that sinus augmentation following autogenous bone graft was significantly greater (mean bone height: 1.6 mm, 95% CI: 0.5-2.7 mm) than for rhBMP-2/ACS (rhBMP-2 at 1.5 mg/mL). In extraction sockets, rhBMP-2/ACS maintained alveolar ridge height while enhancing alveolar ridge width. Safety reports did not represent concerns for the proposed indications. rhBMP-2/ACS appears a promising alternative to autogenous bone grafts for alveolar ridge/maxillary sinus augmentation; dose and carrier optimization may expand its efficacy, use, and clinical application.


In-office vs. operating room procedures for recurrent respiratory papillomatosis.
We conducted a study to analyze hospital and patient costs, outcomes, and patient satisfaction among adults undergoing in-office and operating room procedures for the treatment of recurrent respiratory papillomatosis. Our final study population was made up of 17 patients-1 man and 16 women, aged 30 to 86 years (mean: 62). The mean number of in-office laser procedures per patient was 4.2, and the mean interval between procedures was 5.4 months (although 10 patients underwent only 1 office procedure); the mean number of operating room procedures was 13.5, and the mean interval between procedures was 14.3 months. An equal number of patients reported complications or adverse events with the two types of procedures-5 each. The difference in cost between the office procedure (mean: $3,413.00) and the operating room procedure (mean: $12,382.59) was almost $9,000, but these savings were offset by the fact that the office procedures needed to be performed three times as often. Patients reported slightly more anxiety and discomfort during the office procedures and, overall, they appeared to prefer the operating room procedure. We conclude that office procedures are significantly more cost-effective than operating room procedures, but their use may be limited by patient tolerance and the increased frequency of the procedure.


Change in Overweight from Childhood to Early Adulthood and Risk of Type 2 Diabetes.
Childhood overweight is associated with an increased risk of type 2 diabetes in adulthood. We investigated whether remission of overweight before early adulthood reduces this risk. We conducted a study involving 62,565 Danish men whose weights and heights had been measured at 7 and 13 years of age and in early adulthood (17 to 26 years of age). Overweight was defined in accordance with Centers for Disease Control and Prevention criteria. Data on type 2 diabetes status (at age ≥30 years, 6710 persons) were obtained from a national health registry. Overweight at 7 years of age (3373 of 62,565 men; 5.4%), 13 years of age (3418 of 62,565; 5.5%), or early adulthood (5108 of 62,565; 8.2%) was positively associated with the risk of type 2 diabetes; associations were stronger at older ages at overweight and at younger ages at diagnosis of type 2 diabetes. Men who had had remission of overweight before the age of 13 years had a risk of having type 2 diabetes diagnosed at 30 to 60 years of age that was similar to that among men who had never been overweight (hazard ratio, 0.96; 95% confidence interval [CI], 0.75 to 1.21). As compared with men who had never been overweight, men who had been overweight at 7 and 13 years of age but not during early adulthood had a higher risk of type 2 diabetes (hazard ratio, 1.47; 95% CI, 1.10 to 1.98), but their risk was lower than that among men with persistent overweight (hazard ratio [persistently overweight vs. never overweight], 4.14; 95% CI, 3.57 to 4.79). An increase in body-mass index between 7 years of age and early adulthood was associated with an increased risk of type 2 diabetes, even among men whose weight had been normal at 7 years of age. Childhood overweight at 7 years of age was associated with increased risks of adult type 2 diabetes only if it continued until puberty or later ages. (Funded by the European Union.).


Guidelines on the use of botulinum toxin type A.
Botulinum toxin is available as types A and B. These two different forms need different dosages and hence, the physician needs to be familiar with the formulations. A thorough knowledge of the anatomy and physiology of the muscles in the area to be injected is essential. INDICATIONS FOR BOTULINUM TOXIN: Dynamic wrinkles caused by persistent muscular contractions are the main aesthetic indications for the use of Botulinum toxin. These include forehead lines, glabellar lines, crow's feet, bunny lines, perioral wrinkles, and platysmal bands. Non-aesthetic indications include hyperhidrosis of the palms, soles and axillae. PHYSICIANS' QUALIFICATIONS: Any qualified dermatologist may practice the technique after receiving adequate training in the field. This may be obtained either during post-graduation or at any workshops dedicated to this subject. Botulinum toxin can be administered in the dermatologist's minor procedure room. Detailed counseling with respect to the treatment, desired effects, and longevity of the results should be discussed with the patient. The patient should be given brochures to study and adequate opportunity to seek information. A detailed consent form needs to be completed by the patient. The consent form should include the type of botulinum toxin, longevity expected and possible postoperative complications. Pre- and postoperative photography is recommended. Dosage depends on the area, muscle mass, gender and other factors outlined in these guidelines. It is recommended that beginners should focus on the basic indications in the upper third of the face and that they treat the middle and lower parts of the face only after garnering adequate experience.


Culture of mouse amniotic fluid-derived cells on irradiated STO feeders results in the generation of primitive endoderm cell lines capable of self-renewal in vitro.
The cells present in amniotic fluid (AF) are currently used for prenatal diagnosis of fetal anomalies but are also a potential source of cells for cell therapy. To better characterize putative progenitor cell populations present in AF, we used culture conditions that support self-renewal to determine if these promoted the generation of stable cell lines from AF-derived cells (AFC). Cells isolated from E11.5 mouse were cultured on irradiated STO fibroblast feeder layers in human embryonic germ cell derivation conditions. The cultures grew multicellular epithelial colonies that could be repropagated from single cells. Reverse transcription semiquantitative polymerase chain reaction of established cell lines revealed that they belonged to the extraembryonic endoderm (ExEn) expressing high levels of Gata6, Gata4, Sox17, Foxa2 and Sox7 mRNA. Hierarchical clustering based on the whole transcriptome expression profile of the AFC lines (AFCL) shows significant correlation between transcription profiles of AFCL and blastocyst-derived XEN, an ExEn cell line. In vitro differentiation of AFCL results in the generation of cells expressing albumin and α-fetoprotein (AFP), while intramuscular injection of AFCL into immunodeficient mice produced AFP-positive tumors with primitive endodermal appearance. Hence, E11.5 mouse AF contains cells that efficiently produce XEN lines. These AF-derived XEN lines do not spontaneously differentiate into embryonic-type cells but are phenotypically stable and have the capacity for extensive expansion. The lack of requirement for reprogramming factors to turn AF-derived progenitor cells into stable cell lines capable of massive expansion together with the known ability of ExEn to contribute to embryonic tissue suggests that this cell type may be a candidate for banking for cell therapies.


Ectopic intrathyroidal thymoma: a case report and review.
Ectopic intrathyroidal thymomas are an exceedingly rare clinical entity that can be challenging to diagnose. This report describes a 39-year-old Japanese woman who presented with prominent left-sided thyroid enlargement that was thought to be a dominant thyroid nodule by ultrasound. Two fine-needle aspiration biopsies showed an atypical lymphoid proliferation that was suspicious for although not diagnostic of a low-grade lymphoma. A diagnosis of ectopic intrathyroidal thymoma was made only after appropriate histopathologic assessment of the surgical specimen.


The Impact of Oral Hygiene Maintenance on the Association Between Periodontitis and Osteoporosis: A Nationwide Population-Based Cross Sectional Study.
Both periodontitis and osteoporosis have similar sign of bone resorption in nature. However, the relationship of the severity between these 2 bone-loss diseases is still uncertain.The aim of this study was to investigate the association between the severity of osteoporosis and periodontitis regarding the impact of oral hygiene maintenance. In total, 35,127 osteoporosis patients and 50,498 comparisons were derived from the Longitudinal Health Insurance Database of Taiwan between 2000 and 2010. The population was subdivided into groups according to the different level oral hygiene maintenance and the severity of periodontitis and osteoporosis. The association between osteoporosis and periodontitis was estimated by multinomial logistic regression and rank correlation by Kendall rank correlation test, presented by odds ratio (OR), and 5% confidence intervals (CIs).After controlling the age, sex, and comorbidities, variables in the good oral hygiene maintenance population, we found that periodontitis raised 1.29-fold risk of osteoporosis (95% CI = 1.12-1.49); the risk of osteoporosis was increased with the elevated severity of periodontitis from 1.27 (95% CI = 1.08-1.48) to 1.38 (95% CI = 1.01-1.89). There is a positive correlation between the severity of periodontitis and osteoporosis occurrence in this population (OR = 1.27-1.46; Kendall rank correlation test P = 0.0003). In the poor oral hygiene maintenance population, periodontitis patients had 6.02-fold risk of osteoporosis than those who without periodontitis (95% CI = 4.65-7.81); the risk of osteoporosis was increased with periodontitis severity from 5.96 (95% CI = 4.48-7.92) to 6.37 (95% CI = 3.36-12.1).This result indicated the periodontitis and osteoporosis are conjunctive. The sudden periodontal breakdown of those who with good oral hygiene maintenance might be an indicator for the risk of osteoporosis; if those who were diagnosed as osteoporosis must pay more attention to their periodontal health. Good oral hygiene maintenance might be a crucial factor for preventing the deterioration of osteoporosis progressing; the oral hygiene maintenance plays a significant influence on the association between periodontitis and osteoporosis.


Thermodynamic Origin of Multilayer Structures in Langmuir Polymer Films.
The emergence of polymer-free water surface in a Langmuir polymer film at conditions where a homogeneous coverage has been expected previously is explained on the basis of the surface tensions of polymer and water, γpv and γwv, respectively, as well as the interfacial tension between the two materials, γpw. The polymer molecules considered are 22-residue poly(γ-benzyl-l-glutamate) (PBLG) peptides in α-helical conformation. Values for γpv and γpw derived from MD simulations are consistent with values inferred from experiments considering the emergence of polymer-free surface area for ultrathin films studied using the surface forces apparatus in earlier work. Based on these surface properties, the behavior of individual PBLG peptides at the air-water interface, the dimerization of PBLG peptides, the equilibrium height and width of fibers with given cross section, and the lateral fusion of fibers are described. We show that a prerequisite for the emergence of multilayer structures, which appear locally in domains of sizes of tens to hundreds of micrometers in the considered Langmuir polymer film, is that the condition γpv + γpw - γwv > 0 holds true.


Clinical and virological response to antiretroviral drugs among HIV patients on first-line treatment in Dar-es-Salaam, Tanzania.
In Tanzania, the follow-up on antiretroviral therapy (ART) response is based on clinical outcomes. We investigated virological response and ARV resistance mutations in relation to clinical response in ARV-treated patients. A cross-sectional study of a cohort of 150 patients taking first-line ART in Dar-es-Salaam was conducted. Data were collected using standardized questionnaires and patients' blood samples. HIV viral load testing and genotyping was performed on all viremic samples. Statistical analyses compared clinical responders and non-responders. The median time on ART was 20 months; 71 (47%) patients were ART clinical responders. Clinical non-responders were more likely to have started ART with advanced disease with significantly lower median percentage weight gain (6% versus 20%) with respect to pre-treatment levels. Sixty-one (86%) and 64 (81%) of clinical responders and non-responders, respectively, had undetectable viral loads. Genotyping was successful in 24 (96%) virologically failing patients, among whom 83% had resistance mutations; 67% had dual nucleoside reverse transcriptase inhibitor (NRTI)/non-NRTI (NNRTI) resistance mutations. Seventeen (71%) and 19 (79%) patients had NRTI and NNRTI resistance mutations, respectively, which were related to the ART in use, with no difference between clinical responders and non-responders. The most prevalent subtypes were A and C, found in 9 (38%) and 7 (29%) patients, respectively. The observed virological response was high and did not correlate with clinical response. The prevalence of ARV resistance mutations was high in viraemic patients and was related to the ARV prescribed. We recommend use of viral load monitoring during ART in Tanzania.


Illuminating nanostructured gold electrode: surface plasmons or electron ejection?
Recently, the photoelectric effect in metals in electrolyte environments has aroused the interest of researchers. However, direct evidence for surface plasmons-enhanced electrochemical reactions involving classic outer-sphere reactions of reversible redox couples is seldom reported. We used a surface plasmons-active gold-mushroom-array as a working electrode and observed enhanced faradaic current from ferrocenemethanol following illumination with a xenon lamp. The photoelectric current behaved differently in the presence and absence of oxygen in the solution. The preliminary results are discussed with consideration of the possible mechanisms of electron transfer, although they are very complex, due to the lack of direct evidence. Some consideration was also taken of the research on photoelectrochemical reactions on metallic electrodes.


Genome wide association study of incomplete hippocampal inversion in adolescents.
Incomplete hippocampal inversion (IHI), also called hippocampal malrotation, is an atypical presentation of the hippocampus present in about 20% of healthy individuals. Here we conducted the first genome-wide association study (GWAS) in IHI to elucidate the genetic underpinnings that may contribute to the incomplete inversion during brain development. A total of 1381 subjects contributed to the discovery cohort obtained from the IMAGEN database. The incidence rate of IHI was 26.1%. Loci with P<1e-5 were followed up in a validation cohort comprising 161 subjects from the PING study. Summary statistics from the discovery cohort were used to compute IHI heritability as well as genetic correlations with other traits. A locus on 18q11.2 (rs9952569; OR = 1.999; Z = 5.502; P = 3.755e-8) showed a significant association with the presence of IHI. A functional annotation of the locus implicated genes AQP4 and KCTD1. However, neither this locus nor the other 16 suggestive loci reached a significant p-value in the validation cohort. The h2 estimate was 0.54 (sd: 0.30) and was significant (Z = 1.8; P = 0.036). The top three genetic correlations of IHI were with traits representing either intelligence or education attainment and reached nominal P< = 0.013.


Direct seeding for rice production increased soil erosion and phosphorus runoff losses in subtropical China.
Estimating soil erosion and nutrient losses from surface runoff in paddy fields is essential for the assessment of sustainable rice (Oryza sativa L.) production and water quality protection. Different rice establishment methods have been used in the last three decades in Asia; however, it is still unclear how these methods influence sustainable agriculture and environmental protection in humid areas. The aim of this study was to evaluate the impacts of rice establishment method on soil erosion and phosphorus (P) losses from surface runoff in Hydragric Anthrosols under a subtropical monsoon climate. Total suspended solids (TSS), total P (TP), dissolved P (DP), and particulate P (PP) runoff losses were measured under four rice establishment treatments in 2013 and 2014, including traditional manual transplanting (TT), mechanical transplanting (MT), dry direct seeding (DD), and wet direct seeding (WD). The results showed that the seasonal TSS in the runoff varied from 59.9 to 829.8 kg ha-1 in the two years. Compared with TT, the DD significantly increased the TSS by 481% in 2013 and by 349% in 2014, while the WD significantly increased TSS by 783% in 2013 and by 571% in 2014. In the 2013 and 2014 rice seasons, the field-observed TP runoff losses were from 0.18 to 1.51 kg ha-1. Compared with TT, the DD significantly increased the TP lost by 222% in 2013 and by 197% in 2014, whereas the WD significantly increased the TP lost by 483% in 2013 and by 387% in 2014. However, the TSS and P losses from the MT and TT were similar in both years. The PP runoff losses accounted for 58-77% of the seasonal TP lost. These findings demonstrate that the conversion of traditional manual transplanting to direct seeding increased soil erosion and P runoff losses in subtropical China.


A novel HRM assay for the simultaneous detection and differentiation of eight poxviruses of medical and veterinary importance.
Poxviruses belonging to the Orthopoxvirus, Capripoxvirus and Parapoxvirus genera share common host species and create a challenge for diagnosis. Here, we developed a novel multiplex PCR method for the simultaneous detection and differentiation of eight poxviruses, belonging to three genera: cowpox virus (CPXV) and camelpox virus (CMLV) [genus Orthopoxvirus]; goatpox virus (GTPV), sheeppox virus (SPPV) and lumpy skin disease virus (LSDV) [genus Capripoxvirus]; orf virus (ORFV), pseudocowpox virus (PCPV) and bovine papular stomatitis virus (BPSV) [genus Parapoxvirus]. The assay is based on high-resolution melting curve analysis (HRMCA) of PCR amplicons produced using genus specific primer pairs and dsDNA binding dye. Differences in fragment size and GC content were used as discriminating power. The assay generated three well separated melting regions for each genus and provided additional intra-genus genotyping allowing the differentiation of the eight poxviruses based on amplicon melting temperature. Out of 271 poxviral DNA samples tested: seven CPXV, 25 CMLV, 42 GTPV, 20 SPPV, 120 LSDV, 33 ORFV, 20 PCPV and two BPSV were detected; two samples presented co-infection with CMLV and PCPV. The assay provides a rapid, sensitive, specific and cost-effective method for the detection of pox diseases in a broad range of animal species and humans.


Empiric antibiotic, mechanical ventilation, and central venous catheter duration as potential factors mediating the effect of a checklist prompting intervention on mortality: an exploratory analysis.
Checklists are clinical decision support tools that improve process of care and patient outcomes. We previously demonstrated that prompting critical care physicians to address issues on a daily rounding checklist that were being overlooked reduced utilization of empiric antibiotics and mechanical ventilation, and reduced risk-adjusted mortality and length of stay. We sought to examine the degree to which these process of care improvements explained the observed difference in hospital mortality between the group that received prompting and an unprompted control group. In the medical intensive care unit (MICU) of a tertiary care hospital, we conducted face-to-face prompting of critical care physicians if processes of care on a checklist were being overlooked. A control MICU team used the checklist without prompting. We performed exploratory analyses of the mediating effect of empiric antibiotic, mechanical ventilation, and central venous catheter (CVC)duration on risk-adjusted mortality. One hundred forty prompted group and 125 control group patients were included. One hundred eighty-three patients were exposed to at least one day of empiric antibiotics during MICU admission. Hospital mortality increased as empiric antibiotic duration increased (P<0.001). Prompting was associated with shorter empiric antibiotic duration and lower risk-adjusted mortality in patients receiving empiric antibiotics (OR 0.41, 95% CI 0.18-0.92, P=0.032). When empiric antibiotic duration was added to mortality models, the adjusted OR for the intervention was attenuated from 0.41 to 0.50, suggesting that shorter duration of empiric antibiotics explained 15.2% of the overall benefit of prompting. Evaluation of mechanical ventilation was limited by study size. Accounting for CVC duration changed the intervention effect slightly. In this analysis, some improvement in mortality associated with prompting was explained by shorter empiric antibiotic duration. However, most of the mortality benefit of prompting was unexplained.


Early endovascular experience for treatments of Takayasu's arteritis.
Takayasu's arteritis (TA) is a chronic inflammatory disease that involves the aorta and its major branches; however, only limited data are available on TA in Taiwan. This study presents the clinical features, angiographic findings, and response to treatment of patients with TA at a single institute in Taiwan. A search of the hospital database for ICD9 code 446.7 (Takayasu's disease) between 1990 and 2010 was performed. Seven cases fulfilled the 1990 American College of Rheumatology diagnostic criteria for Takayasu's disease. Angiographic classification was made according to the guidelines of the 1994 International TA Conference in Tokyo. All of our cases were female, and the median age at diagnosis was 27.5 years (range 14-36 years). Four patients had an angiographic classification of type I (57.1%), two were classified as type V (28.6%), and one was classified as type III (14.3%). The most common symptoms/signs were dizziness and vascular bruits. Two patients underwent bypass surgery, four endovascular stenting, and one hybrid bypass with stenting. After a mean follow-up period of 50.3 ± 68.2 months (range 12.3-199.6 months), both the procedure success and survival rates were 100%. There were four restenosis cases (57.1%), one in the surgical bypass group without symptoms (33.3%), and three in the endovascular group (60%), five restenosis in 14 stents (35.7%). Also, these three patients received secondary endovascular procedure for percutaneous transluminal angioplasty or restenting. There have not been any case series reports about treatments of Takayasu's disease in Taiwan to date, based on a search of the PubMed/MEDLINE and Cochrane Library databases. Although endovascular treatment is becoming more prevalent, the restenosis rate is still high, and long-term follow-up and further strategy for restenosis management are the main challenges.


A preformed, topologically stable replication fork. Characterization of leading strand DNA synthesis catalyzed by T7 DNA polymerase and T7 gene 4 protein.
This paper describes the construction of a DNA molecule containing a topologically stable structure that simulates a replication fork. This preformed DNA molecule is a circular duplex of 7.2 X 10(3) base pairs (M13mp6 DNA) from which arises, at a unique BamHI recognition site, a noncomplementary 5'-phosphoryl-terminated single strand of 237 nucleotides (SV40 DNA). This structure has two experimental attributes. 1) Templates for both leading and lagging strand synthesis exist as stable structures prior to any DNA synthesis. 2) DNA synthesis creates a cleavage site for the restriction endonuclease BamHI. Form I of T7 DNA polymerase, alone, catalyzes limited DNA synthesis at the preformed replication fork whereas Form II, alone, polymerizes less than 5 nucleotides. However, when T7 gene 4 protein is present, Form II of T7 DNA polymerase catalyzes rapid and extensive synthesis via a rolling circle mode. Kinetic analysis of this synthesis reveals that the fork moves at a rate of 300 bases/s at 30 degrees C. We conclude that the T7 gene 4 protein requires a single-stranded DNA binding site from which point it translocates to the replication fork where it functions as a helicase. The phage T4 DNA polymerase catalyzes DNA synthesis at this preformed replication fork in the presence of gene 4 protein, but the amount of DNA synthesized is less that 3% of the amount synthesized by the combination of Form II of T7 DNA polymerase and gene 4 protein. We conclude that T7 DNA polymerase and T7 gene 4 protein interact specifically during DNA synthesis at a replication fork.


Inflammatory impact of IFN-γ in CD8+ T cell-mediated lung injury is mediated by both Stat1-dependent and -independent pathways.
Influenza infection results in considerable pulmonary pathology, a significant component of which is mediated by CD8(+) T cell effector functions. To isolate the specific contribution of CD8(+) T cells to lung immunopathology, we utilized a nonviral murine model in which alveolar epithelial cells express an influenza antigen and injury is initiated by adoptive transfer of influenza-specific CD8(+) T cells. We report that IFN-γ production by adoptively transferred influenza-specific CD8(+) T cells is a significant contributor to acute lung injury following influenza antigen recognition, in isolation from its impact on viral clearance. CD8(+) T cell production of IFN-γ enhanced lung epithelial cell expression of chemokines and the subsequent recruitment of inflammatory cells into the airways. Surprisingly, Stat1 deficiency in the adoptive-transfer recipients exacerbated the lung injury that was mediated by the transferred influenza-specific CD8(+) T cells but was still dependent on IFN-γ production by these cells. Loss of Stat1 resulted in sustained activation of Stat3 signaling, dysregulated chemokine expression, and increased infiltration of the airways by inflammatory cells. Taken together, these data identify important roles for IFN-γ signaling and Stat1-independent IFN-γ signaling in regulating CD8(+) T cell-mediated acute lung injury. This is the first study to demonstrate an anti-inflammatory effect of Stat1 on CD8(+) T cell-mediated lung immunopathology without the complication of differences in viral load.


Sequencing and characterisation of an extensive Atlantic salmon (Salmo salar L.) microRNA repertoire.
Atlantic salmon (Salmo salar L.), a member of the family Salmonidae, is a totemic species of ecological and cultural significance that is also economically important in terms of both sports fisheries and aquaculture. These factors have promoted the continuous development of genomic resources for this species, furthering both fundamental and applied research. MicroRNAs (miRNA) are small endogenous non-coding RNA molecules that control spatial and temporal expression of targeted genes through post-transcriptional regulation. While miRNA have been characterised in detail for many other species, this is not yet the case for Atlantic salmon. To identify miRNAs from Atlantic salmon, we constructed whole fish miRNA libraries for 18 individual juveniles (fry, four months post hatch) and characterised them by Illumina high-throughput sequencing (total of 354,505,167 paired-ended reads). We report an extensive and partly novel repertoire of miRNA sequences, comprising 888 miRNA genes (547 unique mature miRNA sequences), quantify their expression levels in basal conditions, examine their homology to miRNAs from other species and identify their predicted target genes. We also identify the location and putative copy number of the miRNA genes in the draft Atlantic salmon reference genome sequence. The Atlantic salmon miRNAs experimentally identified in this study provide a robust large-scale resource for functional genome research in salmonids. There is an opportunity to explore the evolution of salmonid miRNAs following the relatively recent whole genome duplication event in salmonid species and to investigate the role of miRNAs in the regulation of gene expression in particular their contribution to variation in economically and ecologically important traits.


Prolonged herbicide-induced vegetation changes in a regenerating boreal aspen clearcut.
A soil-active herbicide (hexazinone) was applied (0, 2, and 4 kg/ha of active ingredient) in a 3-year-old regenerating boreal Populus tremuloides Michx. (aspen) clearcut to determine its effect on the compositional and structural development of the vegetation. Woody stem densities and plant foliar cover were evaluated prior to and 2, 6, and 17 years after treatment. Herbicide treatment at the 2 and 4 kg/ha rates reduced tree and total woody stem densities relative to the 0 kg/ha level. The 4 kg/ha level reduced stem densities by 27% 17 years after treatment. The primary reductions occurred in Amelanchier alnifolia (Nutt.) Nutt. ex M. Roemer (saskatoon) and Rosa acicularis Lindl. (wild rose); whereas Corylus cornuta Marsh. (beaked hazelnut) and Viburnum edule (Michx.) Raf. (low-bush cranberry) stem densities increased. Notable herbicide-caused foliar cover reductions at the 4 kg/ha level occurred in Eurybia conspicua (Lindl.) Nesom. (showy aster), Mertensia paniculata (Ait.) G. Don. (tall mertensia), Rubus pubescens Raf. (dewberry), and Spiraea betulifolia Pallas (spiraea), but Aralia nudicaulis L. (sarsaparilla), Cornus canadensis L. (bunchberry), and Symphyotrichum ciliolatum (Lindl.) A.&D. Lve (Lindley's aster) increased. Less distinctive but similar changes occurred in the 2 kg/ha treatment. Total plant cover, species richness, and species dominance concentration were similar among treatments. Eight distinctive forest understory-types were recognized among treatments in Year 17. Between the 0 and 4 kg/ha treatments, five understory-types differed in their frequency of occurrence. Hexazinone did not improve the survival of silviculturally planted Picea glauca (Moench) Voss (white spruce) seedlings relative to untreated sites, but the 4 kg/ha treatment level did increase Pinus contorta Dougl. ex Loud. (lodgepole pine) survival from 12 to 34%. Surviving seedlings had significantly greater height and basal diameter growth than those at the 0 kg/ha sites, particularly the 4 kg/ha treatment.


The structure of a thermophilic archaeal virus shows a double-stranded DNA viral capsid type that spans all domains of life.
Of the three domains of life (Eukarya, Bacteria, and Archaea), the least understood is Archaea and its associated viruses. Many Archaea are extremophiles, with species that are capable of growth at some of the highest temperatures and extremes of pH of all known organisms. Phylogenetic rRNA-encoding DNA analysis places many of the hyperthermophilic Archaea (species with an optimum growth > or = 80 degrees C) at the base of the universal tree of life, suggesting that thermophiles were among the first forms of life on earth. Very few viruses have been identified from Archaea as compared to Bacteria and Eukarya. We report here the structure of a hyperthermophilic virus isolated from an archaeal host found in hot springs in Yellowstone National Park. The sequence of the circular double-stranded DNA viral genome shows that it shares little similarity to other known genes in viruses or other organisms. By comparing the tertiary and quaternary structures of the coat protein of this virus with those of a bacterial and an animal virus, we find conformational relationships among all three, suggesting that some viruses may have a common ancestor that precedes the division into three domains of life >3 billion years ago.


Assessing mentalization in psychotherapy: first validation of the Mentalization Imbalances Scale.
The aim of this study was to provide data on the preliminary validation of a clinician-report multidimensional assessment measure of mentalization (Mentalization Imbalances Scale, MIS). A random national sample of psychotherapists (N=190) completed the MIS to identify mentalization imbalances, and the Personality Disorder Checklist to assess the personality disorders (PDs) of randomly selected patients currently in their care. Factor analysis confirmed the presence of six factors that represented different imbalances of mentalization: cognitive, affective, automatic, external, imbalance toward others, and imbalance toward self. We found several significant relationships between patients' mentalization imbalances and personality pathology. Paranoid, schizoid, and schizotypal PDs were predicted by an imbalance toward self, an imbalance the patients shared with histrionic, avoidant, and obsessive compulsive PDs, whereas dependent, borderline, and histrionic PDs were related to an imbalance toward others. Cognitive imbalance was related to schizoid, narcissistic, and obsessive compulsive PDs, whereas affective imbalance predicted antisocial, borderline, narcissistic and histrionic PDs. Automatic imbalance was related to schizotypal, antisocial, and borderline PDs. MIS represents a reliable and valid measure that can help clinicians at understanding patients' specific difficulties of mentalization.


Effects of an irregular bedtime schedule on sleep quality, daytime sleepiness, and fatigue among university students in Taiwan.
An irregular bedtime schedule is a prevalent problem in young adults, and could be a factor detrimentally affecting sleep quality. The goal of the present study was to explore the association between an irregular bedtime schedule and sleep quality, daytime sleepiness, and fatigue among undergraduate students in Taiwan. A total of 160 students underwent a semi-structured interview and completed a survey comprising 4 parts: Pittsburgh Sleep Quality Index (PSQI), Epworth Sleepiness Scale (ESS), Fatigue Severity Scale (FSS), and a rating of irregular bedtime frequency. Participants were grouped into 3 groups in terms of irregular bedtime frequency: low, intermediate, or high according to their 2-week sleep log. To screen for psychological disorders or distress that may have affected responses on the sleep assessment measures, the Chinese health questionnaire-12 (CHQ-12) was also administered. We found an increase in bedtime schedule irregularity to be significantly associated with a decrease in average sleep time per day (Spearman r = -0.22, p = 0.05). Multivariate regression analysis revealed that irregular bedtime frequency and average sleep time per day were correlated with PSQI scores, but not with ESS or FSS scores. A significant positive correlation between irregular bedtime frequency and PSQI scores was evident in the intermediate (partial r = 0.18, p = 0.02) and high (partial r = 0.15, p = 0.05) frequency groups as compared to low frequency group. The results of our study suggest a high prevalence of both an irregular bedtime schedule and insufficient sleep among university students in Taiwan. Students with an irregular bedtime schedule may experience poor sleep quality. We suggest further research that explores the mechanisms involved in an irregular bedtime schedule and the effectiveness of interventions for improving this condition.


Facial nerve paralysis: A case report of rare complication in uncontrolled diabetic patient with mucormycosis.
Mucormycosis is a rare opportunistic aggressive and fatal infection caused by mucor fungus. Seven types of mucormycosis are identified based on the extension and involvement of the lesion, of which the rhino orbital mucormycosis is most common in the head and neck region. Although it is widely spread in nature, clinical cases are rare and observed only in immunocompromised patients and patients with uncontrolled diabetes mellitus. Early symptoms include fever, nasal ulceration or necrosis, periorbital edema or facial swelling, paresthesia and reduced vision. Involvement of cranial nerves although not common, facial nerve palsy is a rare finding. The infection may spread through cribriform plate to the brain resulting in extensive cerebellar infarctions. Timely diagnosis and early recognition of the signs and symptoms, correction of underlying medical disorders, and aggressive medical and surgical intervention are necessary for successful therapeutic outcome.


Combined Use of Unnatural Amino Acids Enables Dual-Color Super-Resolution Imaging of Proteins via Click Chemistry.
Recent advances in optical nanoscopy have brought the imaging resolution to the size of the individual macromolecules, thereby setting stringent requirements for the fluorescent labels. Such requirements are optimally fulfilled by the incorporation of unnatural amino acids (UAAs) in the proteins of interest (POIs), followed by fluorophore conjugation via click chemistry. However, this approach has been limited to single POIs in mammalian cells. Here we solve this problem by incorporating different UAAs in different POIs, which are expressed in independent cell sets. The cells are then fused, thereby combining the different proteins and organelles, and are easily imaged by dual-color super-resolution microscopy. This procedure, which we termed Fuse2Click, is simple, requires only the well-established Amber codon, and allows the use of all previously optimized UAAs and tRNA/RS pairs. This should render it a tool of choice for multicolor click-based imaging.


Simple rules guide dragonfly migration.
Every year billions of butterflies, dragonflies, moths and other insects migrate across continents, and considerable progress has been made in understanding population-level migratory phenomena. However, little is known about destinations and strategies of individual insects. We attached miniaturized radio transmitters (ca 300 mg) to the thoraxes of 14 individual dragonflies (common green darners, Anax junius) and followed them during their autumn migration for up to 12 days, using receiver-equipped Cessna airplanes and ground teams. Green darners exhibited distinct stopover and migration days. On average, they migrated every 2.9+/-0.3 days, and their average net advance was 58+/-11 km in 6.1+/-0.9 days (11.9+/-2.8 km d-1) in a generally southward direction (186+/-52 degrees). They migrated exclusively during the daytime, when wind speeds were less than 25 km h-1, regardless of wind direction, but only after two nights of successively lower temperatures (decrease of 2.1+/-0.6 degrees C in minimum temperature). The migratory patterns and apparent decision rules of green darners are strikingly similar to those proposed for songbirds, and may represent a general migration strategy for long-distance migration of organisms with high self-propelled flight speeds.


MR imaging and PET/CT in diagnosis and management of multiple myeloma.
Multiple myeloma is a common hematologic malignancy among the elderly population. Although there have been many advances in treatment over the past few decades, the overall prognosis for the disease remains poor. Conventional radiography has long been the standard of reference for the imaging of multiple myeloma. However, 10%-20% of patients with multiple myeloma do not have evidence of disease at conventional radiography. There is a growing body of evidence supporting use of magnetic resonance (MR) imaging and 2-[fluorine-18]fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET)/computed tomography (CT) in diagnosis and management of multiple myeloma. MR imaging is useful in detection of bone marrow infiltration, a finding often missed at conventional radiography. FDG PET/CT is especially sensitive for the detection of extramedullary disease and can help detect the metabolically active lesions that often precede evidence of osseous destruction at conventional radiography. MR imaging and FDG PET/CT are useful tools that can provide essential information for diagnosis and management of patients with multiple myeloma. Both modalities allow accurate localization of disease after chemotherapy or autologous stem cell transplantation and can provide important prognostic information that can influence further clinical decision making regarding therapy, particularly when tumor serum markers may be a less reliable indicator of disease burden after repeated treatments.


Fabrication and characterization of optogenetic, multi-strip cardiac muscles.
Cardiac tissue engineering aims to recreate functional tissue constructs similar to the structure and function of the native myocardium. To date, in vitro tissue constructs lack the architectural complexity of a vascular network and the precise motor unit control of muscle fibers. Here, we present a method to construct engineered multi-strip cardiac muscle that simulates the bundle-like architecture of the native myocardium. Densely packed primary myocytes and cardiac fibroblasts were co-cultured with optogenetic, non-excitable cells. The resulting 3D syncytium triggered contraction upon localized blue light illumination to selectively activate and pace the multi-strip cardiac muscles, similar to the activity of pacemaker cells. Acting on a single load, we demonstrated graded force production through light-modulated multi-strip recruitment. These results demonstrate an in vitro platform of optogenetic, multi-strip cardiac muscles that can be used in a wide variety of applications, such as drug discovery, tissue engineering, and bio-hybrid robotic systems.


Lift and drag in intruders moving through hydrostatic granular media at high speeds.
Recently, experiments showed that forces on intruders dragged horizontally through dense, hydrostatic granular packings mainly depend on the local surface orientation and can be seen as the sum of the forces exerted on small surface elements. In order to understand such forces more deeply, we perform a two-dimensional soft-sphere molecular dynamics simulation, on a similar setup, of an intruder dragged through a 50-50 bi-disperse granular packing, with diameters 0.30 and 0.34 cm. We measure, for both circular and half-circle shapes, the forces parallel (drag) and perpendicular (lift) to the drag direction as functions of the drag speed, with V=10.3-309 cm/s, and intruder depths, with D=3.75-37.5 cm. The drag forces on an intruder monotonically increase with V and D, and are larger for the circle. However, the lift force does not depend monotonically on V and D, and this relationship is affected by the shape of the intruder. The vertical force was negative for the half-circle, but for a small range of V and D, we measure positive lift. We find no sign change for the lift on the circle, which is always positive. The explanation for the nonmonotonic dependence is related to the decrease in contacts on the intruder as V increases. This is qualitatively similar to supersonic flow detachment from an obstacle. The detachment picture is supported by simulation measurements of the velocity field around the intruder and force profiles measured on its surface.


Risk-Based Decision Making: A Systematic Scoping Review of Animal Models and a Pilot Study on the Effects of Sleep Deprivation in Rats.
Animals, including humans, frequently make decisions involving risk or uncertainty. Different strategies in these decisions can be advantageous depending the circumstances. Short sleep duration seems to be associated with more risky decisions in humans. Animal models for risk-based decision making can increase mechanistic understanding, but very little data is available concerning the effects of sleep. We combined primary- and meta-research to explore the relationship between sleep and risk-based decision making in animals. Our first objective was to create an overview of the available animal models for risky decision making. We performed a systematic scoping review. Our searches in Pubmed and Psychinfo retrieved 712 references, of which 235 were included. Animal models for risk-based decision making have been described for rodents, non-human primates, birds, pigs and honey-bees. We discuss task designs and model validity. Our second objective was to apply this knowledge and perform a pilot study on the effect of sleep deprivation. We trained and tested male Wistar rats on a probability discounting task; a "safe" lever always resulted in 1 reward, a "risky" lever resulted in 4 or no rewards. Rats adapted their preferences to variations in reward probabilities (p < 0.001), but 12 h of sleep deprivation during the light phase did not clearly alter risk preference (p = 0.21).


Nitric oxide production during cerebral ischemia and reperfusion in eNOS- and nNOS-knockout mice.
The purpose of this study was to clarify the kinetics of nitric oxide (NO) induced by either endothelial NO synthase (eNOS) or neuronal NO synthase (nNOS) after transient global forebrain ischemia. We investigated NO production and ischemic changes to hippocampal CA1 neurons in eNOS knockout (-/-) mice and nNOS (-/-) mice during cerebral ischemia and reperfusion. NO production was continuously monitored by in vivo microdialysis. Global forebrain ischemia was produced by occlusion of both common carotid arteries for 10 minutes. Levels of nitrite (NO(2)(-)) and nitrate (NO(3)(-)), as NO metabolites, in dialysate were determined using the Griess reaction. Two hours after the start of reperfusion, animals were perfused with 4% paraformaldehyde. Hippocampal CA1 neurons were divided into three phases (severely ischemic, moderately ischemic, surviving), and the ratio of surviving neurons to degenerated neurons was calculated as the survival rate. The relative cerebral blood flow (rCBF) was significantly higher in nNOS (-/-) mice than in control mice after reperfusion. Levels of NO(3)(-) were significantly lower in eNOS (-/-) mice and nNOS (-/-) mice than in control mice during ischemia and reperfusion. NO(3)(-) levels were significantly lower in nNOS (-/-) mice than in eNOS (-/-) mice after the start of reperfusion. Survival rate tended to be higher in nNOS (-/-) mice than in control mice, but not significantly. These in vivo data suggest that NO production in the striatum after reperfusion is closely related to activities of both nNOS and eNOS, and is mainly related to nNOS following reperfusion.


Changes in bone structure and mass with advancing age in the male C57BL/6J mouse.
To determine whether the mouse loses bone with aging and whether the changes mimic those observed in human aging, we examined the changes in the tibial metaphysis and diaphysis in the male C57BL/6J mouse over its life span using microcomputed tomography (microCT). Cancellous bone volume fraction (BV/TV) decreased 60% between 6 weeks and 24 months of age. Loss was characterized by decreased trabecular number (Tb.N), increased trabecular spacing (Tb.Sp), and decreased connectivity. Anisotropy decreased while the structure model index increased with age. Cortical bone thickness increased between 6 weeks and 6 months of age and then decreased continuously to 24 months (-12%). Cortical bone area (Ct.Ar) remained constant between 6 and 24 months. Fat-free weight reached a peak at 12 months and gradually declined to 24 months. Total mass lost between 12 and 24 months reached 10%. Overall, the age-related changes in skeletal mass and architecture in the mouse were remarkably similar to those seen in human aging. Furthermore, the rapid early loss of cancellous bone suggests that bone loss is not just associated with old age in the mouse but rather occurs as a continuum from early growth. We conclude that the C57BL/6J male mouse maybe a useful model to study at least some aspects of age-related bone loss in humans.


Microarray R-based analysis of complex lysate experiments with MIRACLE.
Reverse-phase protein arrays (RPPAs) allow sensitive quantification of relative protein abundance in thousands of samples in parallel. Typical challenges involved in this technology are antibody selection, sample preparation and optimization of staining conditions. The issue of combining effective sample management and data analysis, however, has been widely neglected. This motivated us to develop MIRACLE, a comprehensive and user-friendly web application bridging the gap between spotting and array analysis by conveniently keeping track of sample information. Data processing includes correction of staining bias, estimation of protein concentration from response curves, normalization for total protein amount per sample and statistical evaluation. Established analysis methods have been integrated with MIRACLE, offering experimental scientists an end-to-end solution for sample management and for carrying out data analysis. In addition, experienced users have the possibility to export data to R for more complex analyses. MIRACLE thus has the potential to further spread utilization of RPPAs as an emerging technology for high-throughput protein analysis. Project URL: http://www.nanocan.org/miracle/. Supplementary data are available at Bioinformatics online.


Are value of information methods ready for prime time? An application to alternative treatment strategies for NSTEMI patients.
The use of value of information methods to inform trial design has been widely advocated but there have been few empirical applications of these methods and there is little evidence they are widely used in decision making. This study considers the usefulness of value of information models in the context of a real clinical decision problem relating to alternative diagnostic strategies for patients with a recent non-ST elevated myocardial infarction. A pretrial economic model is constructed to consider the cost-effectiveness of two competing strategies: coronary angiography alone or in conjunction with fractional flow reserve measurement. A closed-form solution to the expected benefits of information is used with optimal sample size estimated for a range of models reflecting increasingly realistic assumptions and alternative decision contexts. Fractional flow reserve measurement is expected to be cost-effective with an incremental cost-effectiveness ratio of GBP 1,621, however, there is considerable uncertainty in this estimate and consequently a large expected value to reducing this uncertainty via a trial. The recommended sample size is strongly affected by the reality of the assumptions of the expected value of information (EVI) model and the decision context. Value of information models can provide a simple and flexible approach to clinical trial design and are more consistent with the constraints and objectives of the healthcare system than traditional frequentist approaches. However, the variation in sample size estimates demonstrates that it is essential that appropriate model parameters and decision contexts are used in their application.


Twin analyses of chronic fatigue in a Swedish national sample.
Chronic fatigue has infrequently been studied in twins. Data from twin studies can inform clinical and research approaches to the management and etiology of human complex traits. The authors obtained telephone interview data on current chronic fatigue from 31406 individuals twins in the Swedish Twin Registry (aged 42-64 years, 75.68% response rate), from both members of 12407 pairs and from one member of 6592 pairs. Of the complete pairs, 3269 pairs were monozygotic, 9010 pairs dizygotic, and 128 pairs of unknown zygosity. Structural equation twin modeling was used to estimate the latent genetic architecture of varying definitions of fatiguing illness. Estimates of additive genetic effects, shared environmental effects, and individual-specific environmental effects were similar in males and females. No definition of current fatiguing illness (ranging from any fatigue to CFS-like illness) was strikingly distinctive. Individual-specific effects were the predominant source of variation, followed by modest genetic influences. We could not exclude a small but conceptually important contribution of shared environmental effects. Current fatiguing illness appears to be a complex trait resulting from both environmental and genetic sources of variation without pronounced differences by gender.


Factors associated with consistent condom use and STIs among foreign female entertainment workers: results from a cross-sectional survey in Singapore.
We assessed the prevalence of consistent condom use and laboratory-confirmed STIs among foreign female entertainment workers (FEWs) who engaged in paid or casual sex in Singapore and the factors associated with these characteristics. A cross-sectional survey, using time-location sampling, was conducted on 220 FEWs (115 Vietnamese and 105 Thai) in 2015. For multivariable analysis, we used a mixed-effects Poisson regression model with backward stepwise approach to account for clustering by venue and to obtain the adjusted prevalence ratios (aPRs) of factors associated with consistent condom use and STI positivity, respectively. Consistent condom use with paid or casual partners in the past month was 39.6% and 36.2% for vaginal and oral sex, respectively. The prevalence of STI (cervical chlamydia, cervical gonorrhoea or pharyngeal gonorrhoea) positivity was 13.6%. In multivariable analysis, consistent condom use for both vaginal and oral sex increased with high self-perceived risk of getting STI/HIV (vaginal: aPR 2.09; 95% CI 1.26 to 3.46; oral: aPR 2.41; 95% CI 1.23 to 4.69) and condom negotiation (vaginal: aPR 3.74; 95% CI 2.07 to 6.75; oral: aPR 2.81; 95% CI 1.51 to 5.26). STI positivity decreased with consistent condom use for vaginal sex (aPR 0.22; 95% CI 0.07 to 0.75) and increased with number of sexual partners (aPR 1.43; 95% CI 1.07 to 1.92). In addition to advocating for policy decisions to provide a safer work environment, behavioural interventions on condom negotiation skills and condom use and biomedical interventions on STI/HIV testing and treatment interventions are needed among the FEWs in Singapore. NCT02780986; pre-results.


Clinical and epidemiologic features of SARS-CoV-2 in dogs and cats compiled through national surveillance in the United States.
To characterize clinical and epidemiologic features of SARS-CoV-2 in companion animals detected through both passive and active surveillance in the US. 204 companion animals (109 cats, 95 dogs) across 33 states with confirmed SARS-CoV-2 infections between March 2020 and December 2021. Public health officials, animal health officials, and academic researchers investigating zoonotic SARS-CoV-2 transmission events reported clinical, laboratory, and epidemiologic information through a standardized One Health surveillance process developed by the CDC and partners. Among dogs and cats identified through passive surveillance, 94% (n = 87) had reported exposure to a person with COVID-19 before infection. Clinical signs of illness were present in 74% of pets identified through passive surveillance and 27% of pets identified through active surveillance. Duration of illness in pets averaged 15 days in cats and 12 days in dogs. The average time between human and pet onset of illness was 10 days. Viral nucleic acid was first detected at 3 days after exposure in both cats and dogs. Antibodies were detected starting 5 days after exposure, and titers were highest at 9 days in cats and 14 days in dogs. Results of the present study supported that cats and dogs primarily become infected with SARS-CoV-2 following exposure to a person with COVID-19, most often their owners. Case investigation and surveillance that include both people and animals are necessary to understand transmission dynamics and viral evolution of zoonotic diseases like SARS-CoV-2.


Follicular maturation, luteinization and first meiotic division in oocytes after inhibiting mitochondrial function in mice with chloramphenicol.
A significant number of diploid oocytes is ovulated from adult NMRI/Han mice treated with high doses of gonadotrophins. This inhibition of the first meiotic division is very likely caused by an altered communication between the germ cell and the surrounding somatic cells leading to a failure of the endocrine control of meiosis. The present study examined the role of mitochondria in follicular development, oocyte maturation and chromosomal segregation during first meiotic division in NMRI/Han mice. To affect mitochondrial function during the late phase of follicular maturation, chloramphenicol, a potent inhibitor of mitochondrial peptidyl transferase, was used. Adult mice were treated with chloramphenicol (CAM; 18.8 or 37.5 mg/kg b.w.) at different times after the pregnant mare serum injection. The results revealed that CAM inhibited the characteristic increase of ovarian weight, reduced the number of oocytes ovulated per female, lowered the progesterone concentration in the postovulatory ovary and increased the incidence of ovulated diploid oocytes. It was concluded that an irregular mitochondrial function may affect normal follicular development and oocyte maturation, and potentially interferes with the order chromosome segregation during the first meiotic division.


Indirect vs Direct Voice Therapy for Children With Vocal Nodules: A Randomized Clinical Trial.
Benign vocal fold nodules affect 12% to 22% of the pediatric population, and 95% of otolaryngologists recommend voice therapy as treatment. However, no randomized clinical trials that we are aware of have shown its benefits. To determine the impact of voice therapy in children with vocal fold nodules according to pretherapy and posttherapy scores on the Pediatric Voice-Related Quality of Life (PVRQOL) survey; secondary objectives included changes in phonatory parameters. For this multicenter randomized clinical trial, 114 children ages 6 to 10 years with vocal fold nodules, PVRQOL scores less than 87.5, and dysphonia for longer than 12 weeks were recruited from outpatient voice and speech clinics. This age range was identified because these patients have not experienced pubertal changes of the larynx, tolerate stroboscopy, and cooperate with voice therapy. Participants were blinded to treatment arm. Participants received either indirect or direct therapy for 8 to 12 weeks. Indirect therapy focused on education and discussion of voice principles, while direct treatment used the stimulus, response, antecedent paradigm. The primary outcome measure was PVRQOL score change before and after treatment. Secondary phonatory measures were also compared. Overall, 114 children were recruited for study (mean [SD] age, 8 [1.4] years; 83 males [73%]); with 57 randomized to receive either indirect or direct therapy. Both direct and indirect therapy approaches showed significant differences in PVRQOL scores pretherapy to posttherapy. The mean increase in PVRQOL score for direct therapy was 19.2, and 14.7 for indirect therapy (difference, 4.5; 95.3% CI, -10.8 to 19.8). Of 44 participants in the direct therapy group, 27 (61%) achieved a clinically meaningful PVRQOL improvement, compared with 26 of 49 (53%) for indirect therapy (difference, 8%; 95% CI, -12 to 28). Post hoc stratification showed robust effects in the direct therapy group for older children (Cohen d = 0.50) and the latter two-thirds of participants (Cohen d = 0.46). Vocal fold nodules reduced in size in 31% (22 of 70) and completely resolved in 11% (8 of 70) of participants who consented to a second set of images after going through the recruitment process. Both direct and indirect voice therapy improved voice-related quality of life in children with vocal fold nodules, although there was no significant difference between approaches. Future studies may focus upon which voice therapy approaches are effective in treating age-defined populations. clinicaltrials.gov Identifier: NCT01255735.


Long-term effects of bereavement and caregiver intervention on dementia caregiver depressive symptoms.
The purpose of this study was to examine the joint effects of bereavement and caregiver intervention on caregiver depressive symptoms. Alzheimer's caregivers from a randomized trial of an enhanced caregiver support intervention versus usual care who had experienced the death of their spouse (n = 254) were repeatedly assessed with the Geriatric Depression Scale prior to and following bereavement. Random effects regression growth curve analyses examined the effects of treatment group and bereavement while controlling for other variables. The death of the care recipient led to reductions in depressive symptoms for both caregiving groups. Enhanced support intervention led to lower depressive symptoms compared with controls both before and after bereavement. Post-bereavement group differences were stronger for caregivers of spouses who did not previously experience a nursing home placement. These caregivers maintained these differences for more than 1 year after bereavement. Caregivers who received the enhanced support intervention were more likely to show long-term patterns of fewer depressive symptoms before and after bereavement, suggesting resilience, whereas control caregivers were more likely to show chronic depressive symptoms before and after the death of their spouse. Caregiver intervention has the potential to alter the long-term course of the caregiving career. Such clinical strategies may also protect caregivers against chronic depressive symptoms that would otherwise persist long after caregiving ends.


Laser punch-out for acne scars.
Patients with acne scars want smooth facial skin. However, achieving this is difficult with dermabrasion or chemical peeling. Nor can acne scars be covered with cosmetics, due to their ice-picked or cobblestone appearance. Laser resurfacing is more effective and safer than other conventional methods due to its precision with depth control and variable methods of surface cutting. Even depth resurfacing with a laser shows unsatisfactory results, therefore, for the deep-sited acne scar the cutting methods have to be changed according to the depth and pattern of the scar. For 2 years, starting in January 1996, we treated 71 patients with a high-powered CO2 laser (Ultrapulse). Different resurfacing methods were applied according to the depth and pattern of the scars. For mild depressed scars, even depth resurfacing was done. For moderate-depth acne scars, the shoulder technique was also used. For the deepest and ice-picked scars, the laser punch-out was combined. Laser resurfacing was carried out at 300-500 mJ, with two to five passes. Laser punch-out was done at 500 mJ, with three to seven continuous passes on the ice-picked scar. From the pathologic findings of acne scars showing that there was thick intradermal scar, we knew that laser punch-out was necessary for improvement of acne scars. Depth-wide, the ice-picked scars improved by over 80% and the sharp demarcated margin of the acne scar faded out. Most of the patients with acne scars were satisfied with laser resurfacing. Only six patients had a second laser treatment, with an interval of 12 months. There were no hypertrophic scars after laser resurfacing, but erythema lasted for 3-12 months. Patients taking oral retinoic acid were not contraindicated for laser resurfacing but required special caution because they had atrophic skin and delayed wound healing. Laser resurfacing is the most versatile method for acne scars, with a high-powered CO2 laser. The laser punch-out method is better than even depth resurfacing for improving deep acne scars and can be combined with the shoulder technique or even depth resurfacing according to the type of acne scar.


Sedentary time, physical activity and compliance with IOM recommendations in young children at childcare.
The aim of this study was to report patterns of sitting, standing and physical activity (PA) and compliance with Institute of Medicine (IOM) recommendations for sedentary behavior (SB) and PA among children aged 1 to 5 years at childcare, and examine sociodemographic variations. Sitting, standing and PA time was assessed using an activPAL inclinometer over a period of 1 to 5 days in 301 children (49% boys; mean age = 3.7 ± 1.0 years) across 11 childcare services in Illawarra, NSW, Australia. Breaks and bouts of sitting and standing were calculated and categorized. Height and weight were assessed and parents completed a demographic survey. Differences by sex, age category (< 3 vs ≥ 3 years), weight status and SES were examined. Children spent 48.4% of their time at childcare sitting, 32.5% standing, and 19.1% in PA. Boys spent significantly more time in PA compared to girls (20.8% vs 17.7%; P = 0.003). Toddlers (< 3 years) spent significantly more time in PA compared to preschoolers (≥ 3 years) (22.2% vs 18.3%; P < 0.001). Children who were underweight spent significantly more time sitting compared with their overweight peers (52.4% vs 46.8%; P = 0.003). 56% and 16% of children met the IOM SB and PA recommendations, respectively. Girls (odds ratio [OR]; 95%CI = 0.26; 0.13 to 0.55) and preschoolers (0.16; 0.07 to 0.38) were less likely to meet the IOM PA recommendation compared to boys and toddlers. Young children spent ~ 50% of their time at childcare sitting. Girls and preschoolers sit more and are less likely to meet PA recommendations, making them important groups to target in future interventions.


Specific IgG subclass antibody levels and phagocytosis of serotype 14 pneumococcus following immunization.
Complement and specific antibody directed against capsular polysaccharide are necessary for efficient phagocytosis of pneumococci. In normal adults, specific antibody to pneumococci is predominantly of the IgG2 subclass. However, the role of IgG2 in bacterial clearance is debatable. We therefore decided to investigate the relationship between specific IgG subclass antibody levels and phagocytosis of serotype 14 pneumococcus, before and after immunization with a pneumococcal capsular polysaccharide vaccine. Specific IgG subclass antibody was measured by an ELISA technique and the effect of serum on phagocytosis of radiolabelled pneumococci by normal polymorphs was determined. We found that in the presence of complement, phagocytosis correlated significantly with both specific IgG1 and IgG2 antibody titres (r = 0.547, P = 0.002 and r = 0.464, P = 0.009, respectively). However, in decomplemented sera, the correlation with IgG1 antibody was lost, whereas that with IgG2 antibody was strengthened (r = 0.641, P = less than 0.001). The possibility that IgG2 binds to receptors on polymorphs should be considered.


Incidence and clinical characteristics of thyroid cancer in prospective series of individuals with Cowden and Cowden-like syndrome characterized by germline PTEN, SDH, or KLLN alterations.
Thyroid cancer is believed to be an important component of Cowden syndrome (CS). Germline PTEN and SDHx mutations and KLLN epimutation cause CS and CS-like phenotypes. Despite the established association, little is known about the incidence and clinical features of thyroid cancer found in CS/CS-like patients. The aim of the study was to compare incidence, clinical, and histological characteristics of epithelial thyroid cancers in CS/CS-like individuals, in the context of PTEN, SDHx, and KLLN status. The study encompassed a 5-yr, multicenter, prospective accrual of 2723 CS and CS-like patients, all of whom had comprehensive PTEN analysis. SDHx mutation analysis occurred in those without PTEN mutations/variations and elevated manganese superoxide dismutase (MnSOD) levels. KLLN epimutation analysis was performed in the subset without any PTEN or SDHx mutation/deletion/ variant/polymorphism. Gene-specific thyroid cancer histologies, demographic and clinical information, and adjusted standardized incidence rates were studied. Of 2723 CS/CS-like patients, 664 had thyroid cancer. Standardized incidence rates for thyroid cancer were 72 [95% confidence interval (CI), 51-99; P < 0.001] for pathogenic PTEN mutations, 63 (95% CI, 42-92; P < 0.001) for SDHx variants, and 45 (95% CI, 26-73; P < 0.001) for KLLN epimutations. All six (16.7%) diagnosed under age 18 yr carried pathogenic PTEN mutations. Follicular thyroid cancer was overrepresented in PTEN mutation-positive cases compared to those with SDHx and KLLN alterations. PTEN frameshift mutations were found in 31% of patients with thyroid cancer compared to 17% in those without thyroid cancer. CS/CS-like patients have elevated risks of follicular thyroid cancer due to PTEN pathogenic mutations and of papillary thyroid cancer from SDHx and KLLN alterations. Children presenting with thyroid cancer should be tested for PTEN mutations.


Comparison of the construction of unmarked deletion mutations in Mycobacterium smegmatis, Mycobacterium bovis bacillus Calmette-Guérin, and Mycobacterium tuberculosis H37Rv by allelic exchange.
Until recently, genetic analysis of Mycobacterium tuberculosis, the causative agent of tuberculosis, was hindered by a lack of methods for gene disruptions and allelic exchange. Several groups have described different methods for disrupting genes marked with antibiotic resistance determinants in the slow-growing organisms Mycobacterium bovis bacillus Calmette-Guérin (BCG) and M. tuberculosis. In this study, we described the first report of using a mycobacterial suicidal plasmid bearing the counterselectable marker sacB for the allelic exchange of unmarked deletion mutations in the chromosomes of two substrains of M. bovis BCG and M. tuberculosis H37Rv. In addition, our comparison of the recombination frequencies in these two slow-growing species and that of the fast-growing organism Mycobacterium smegmatis suggests that the homologous recombination machinery of the three species is equally efficient. The mutants constructed here have deletions in the lysA gene, encoding meso-diaminopimelate decarboxylase, an enzyme catalyzing the last step in lysine biosynthesis. We observed striking differences in the lysine auxotrophic phenotypes of these three species of mycobacteria. The M. smegmatis mutant can grow on lysine-supplemented defined medium or complex rich medium, while the BCG mutants grow only on lysine-supplemented defined medium and are unable to form colonies on complex rich medium. The M. tuberculosis lysine auxotroph requires 25-fold more lysine on defined medium than do the other mutants and is dependent upon the detergent Tween 80. The mutants described in this work are potential vaccine candidates and can also be used for studies of cell wall biosynthesis and amino acid metabolism.


Housing trajectories and the risk of homelessness among new mental health service users: Protocol for the AMONT study.
High rates of mental illness among individuals experiencing homelessness, as well as documented contacts with psychiatric services prior to episodes of homelessness, indicate that early intervention could play a key role in homelessness prevention. Decision-makers and clinical teams need longitudinal data on housing trajectories following initial contact with psychiatric services, as well as on predictors of risk of housing instability and homelessness. This paper describes the AMONT study, a mixed-methods naturalistic longitudinal cohort study of individuals identified as new psychiatric service users in seven clinical sites across the province of Québec (Canada). The goal of AMONT is to examine the housing situations of individuals over 36 months after their initial contact with psychiatric services, and to identify environmental and individual correlates and predictors of housing outcomes. Participants complete a broad battery of instruments at baseline and follow-up assessments after 24 and 36 months. We explore housing stability following an initial episode of psychiatric service use from the perspective of service users, family members, and service providers, through qualitative interviews. The findings from the AMONT study will yield a better understanding of the residential pathways of individuals with mental illness, from their first contact with psychiatric services and for 3 years subsequently. This will inform service providers, decision-makers and managers on the specific housing concerns and issues that affect first-time mental health service users. This in turn can lead to the development and implementation of evidence-informed practices and policies that aim to prevent instability and homelessness.