Advances in immunostains used in Mohs surgery.
In recent years, Mohs micrographic surgery (MMS) has become a widely utilized method of removal for a variety of cutaneous neoplasms. Certain clinical scenarios, however, make it difficult to visualize residual tumor cells, potentially decreasing the efficacy of the Mohs procedure. Immunohistochemical (IHC) stains are now available and are being utilized to delineate cells of interest intraoperatively when routinely stained slides are equivocal. While useful, IHC stains have not gained wide acceptance as an adjunct to MMS, particularly due to increased processing time, cost and workload required. There have been multiple recent advances, however, in the utilization of IHC stains in MMS. In this article, the authors discuss recent advances in IHC stains used in MMS for the treatment of melanoma as well as nonmelanoma skin cancers, potentially making their routine use in select cases more feasible.


Bridging process-based and empirical approaches to modeling tree growth.
The gulf between process-based and empirical approaches to modeling tree growth may be bridged, in part, by the use of a common model. To this end, we have formulated a process-based model of tree growth that can be fitted and applied in an empirical mode. The growth model is grounded in pipe model theory and an optimal control model of crown development. Together, the pipe model and the optimal control model provide a framework for expressing the components of tree biomass in terms of three standard inventory variables: tree height, crown height and stem cross-sectional area. Growth rates of the inventory variables and the components of biomass are formulated from a carbon balance. Fundamentally, the parameters of the model comprise physiological rates and morphological ratios. In principle, the values of these parameters may be estimated by lower-level process models. Alternatively, the physiological and morphological parameters combine, under reasonable assumptions, into a set of aggregate parameters, whose values can be estimated from inventory data with a statistical fitting procedure.


Sustainable food systems for optimal planetary health.
Sustainable food systems are an important component of a planetary health strategy to reduce the threat of infectious disease, minimize environmental footprint and promote nutrition. Human population trends and dietary transition have led to growing demand for food and increasing production and consumption of meat, amid declining availability of arable land and water. The intensification of livestock production has serious environmental and infectious disease impacts. Land clearing for agriculture alters ecosystems, increases human-wildlife interactions and leads to disease proliferation. Context-specific interventions should be evaluated towards optimizing nutrition resilience, minimizing environmental footprint and reducing animal and human disease risk.


Association of Acquired and Heritable Factors With Intergenerational Differences in Age at Symptomatic Onset of Alzheimer Disease Between Offspring and Parents With Dementia.
Acquired and heritable traits are associated with dementia risk; however, how these traits are associated with age at symptomatic onset (AAO) of Alzheimer disease (AD) is unknown. Identifying the associations of acquired and heritable factors with variability in intergenerational AAO of AD could facilitate diagnosis, assessment, and counseling of the offspring of parents with AD. To quantify the associations of acquired and heritable factors with intergenerational differences in AAO of AD. This nested cohort study used data from the Knight Alzheimer Disease Research Center that included community-dwelling participants with symptomatic AD, parental history of dementia, and available DNA data who were enrolled in prospective studies of memory and aging from September 1, 2005, to August 31, 2016. Clinical, biomarker, and genetic data were extracted on January 17, 2017, and data analyses were conducted from July 1, 2017, to August 20, 2019. The associations of acquired (ie, years of education; body mass index; history of cardiovascular disease, hypertension, hypercholesterolemia, diabetes, active depression within 2 years, traumatic brain injury, tobacco use, and unhealthy alcohol use; and retrospective determination of AAO) and heritable factors (ie, ethnicity/race, paternal or maternal inheritance, parental history of early-onset dementia, APOE ε4 allele status, and AD polygenic risk scores) to intergenerational difference in AAO of AD were quantified using stepwise forward multivariable regression. Missense or frameshift variants within genes associated with AD pathogenesis were screened using whole-exome sequencing. There were 164 participants with symptomatic AD, known parental history of dementia, and available DNA data (mean [SD] age, 70.9 [8.3] years; 90 [54.9%] women) included in this study. Offspring were diagnosed with symptomatic AD a mean (SD) 6.1 (10.7) years earlier than their parents (P < .001). The adjusted R2 for measured acquired and heritable factors for intergenerational difference in AAO of AD was 0.29 (F8,155 = 9.13; P < .001). Paternal (β = -9.52 [95% CI, -13.79 to -5.25]) and maternal (β = -6.68 [95% CI, -11.61 to -1.75]) history of dementia, more years of education (β = -0.58 [95% CI -1.08 to -0.09]), and retrospective determination of AAO (β = -3.46 [95% CI, -6.40 to -0.52]) were associated with earlier-than-expected intergenerational difference in AAO of AD. Parental history of early-onset dementia (β = 21.30 [95% CI, 15.01 to 27.59]), presence of 1 APOE ε4 allele (β = 5.00 [95% CI, 2.11 to 7.88]), and history of hypertension (β = 3.81 [95% CI, 0.88 to 6.74]) were associated with later-than-expected intergenerational difference in AAO of AD. Missense or frameshift variants within genes associated with AD pathogenesis were more common in participants with the greatest unexplained variability in intergenerational AAO of AD (19 of 48 participants [39.6%] vs 26 of 116 participants [22.4%]; P = .03). Acquired and heritable factors were associated with a substantial proportion of variability in intergenerational AAO of AD. Variants in genes associated with AD pathogenesis may contribute to unexplained variability, justifying further study.


Relationship between haemolysis production and resistance to fluoroquinolones among clinical isolates of Escherichia coli.
The activities of ampicillin, amoxycillin-clavulanic acid, gentamicin, tetracycline, nalidixic acid, ciprofloxacin, pefloxacin and trovafloxacin against 207 consecutive clinical isolates of Escherichia coli were determined. Fifty-six (27.3%) isolates were haemolytic. The percentages of resistance to quinolones and tetracycline, but not to other agents, among haemolytic isolates were significantly lower (P < 0.05) than among non-haemolytic isolates. Ciprofloxacin-resistant mutants obtained from ciprofloxacin-susceptible haemolytic isolates still produced haemolysis. It is concluded that most quinolone-resistant clinical isolates of E. coli are non-haemolytic, although haemolysis is produced by quinolone-resistant mutants derived from haemolytic quinolone-susceptible strains.


An Algorithm to Evaluate Suspected Lung Metastases in Patients with HPV-Associated Oropharyngeal Cancer.
Distinguishing between distantly metastatic and metachronous lung primary carcinoma is challenging for patients with a history of head and neck cancer. There are implications for registry data, prognosis and related counseling, and management options, including eligibility for precision oncology trials. Patients with human papillomavirus (HPV)-associated oropharyngeal squamous cell carcinoma who were treated under a uniform clinical protocol and achieved a complete response were identified in a single-institution prospective head and neck cancer epidemiology database (n = 205). Fifteen patients presented with pulmonary nodule(s) after completion of therapy. We describe our algorithm for the evaluation of these patients, including histopathology, p16 immunohistochemistry, and HPV in situ hybridization.


Knowledge, attitudes, and practices related to TB among the general population of Ethiopia: Findings from a national cross-sectional survey.
Ethiopia is among the high-burden countries for tuberculosis (TB), TB/HIV, and drug-resistant TB. The aim of this nationwide study was to better understand TB-related knowledge, attitudes, and practices (KAPs) and generate evidence for policy and decision-making. We conducted a cross-sectional TB KAP survey in seven regions and two city administrations of Ethiopia. Eighty kebeles (wards) and 40 health centers were randomly selected for the study. Using systematic sampling, 22 households and 11 TB patients were enrolled from each selected village and health center, respectively. Variables with a value of p = < 0.25 were included in the model for logistic regression analysis. Of 3,503 participants, 884 (24.4%), 836 (24.1%), and 1,783 (51.5%) were TB patients, families of TB patients, and the general population, respectively. The mean age was 34.3 years, and 50% were women. Forty-six percent were heads of households, 32.1% were illiterate, 20.3% were farmers, and 19.8% were from the lowest quintile. The majority (95.5%) had heard about TB, but only 25.8% knew that TB is caused by bacteria. Cough or sneezing was reported as the commonest means of TB transmission. The majority (85.3%) knew that TB could be cured. Men, better-educated people, and TB patients and their families have higher knowledge scores. Of 2,483 participants, 96% reported that they would go to public health facilities if they developed TB symptoms. Most Ethiopians have a high level of awareness about TB and seek care in public health facilities, and communities are generally supportive. Inadequate knowledge about TB transmission, limited engagement of community health workers, and low preference for using community health workers were the key challenges. Given misconceptions about TB's causes, low preference for use of community health workers, and inadequate engagement, targeted health education interventions are required to improve TB services.


Innate host response in primary human hepatocytes with hepatitis C virus infection.
The interaction between hepatitis C virus (HCV) and innate antiviral defense systems in primary human hepatocytes is not well understood. The objective of this study is to examine how primary human hepatocytes response to HCV infection. An infectious HCV isolate JFH1 was used to infect isolated primary human hepatocytes. HCV RNA or NS5A protein in the cells was detected by real-time PCR or immunofluorescence staining respectively. Apoptosis was examined with flow cytometry. Mechanisms of HCV-induced IFN-β expression and apoptosis were determined. Primary human hepatocytes were susceptible to JFH1 virus and released infectious virus. IFN-α inhibited viral RNA replication in the cells. IFN-β and interferon-stimulated genes were induced in the cells during acute infection. HCV infection induced apoptosis of primary human hepatocytes through the TRAIL-mediated pathway. Silencing RIG-I expression in primary human hepatocytes inhibited IFN-β and TRAIL expression and blocked apoptosis of the cells, which facilitated viral RNA replication in the cells. Moreover, HCV NS34A protein inhibited viral induced IFN-β expression in primary human hepatocytes. Innate host response is intact in HCV-infected primary human hepatocytes. RIG-I plays a key role in the induction of IFN and TRAIL by viruses and apoptosis of primary human hepatocytes via activation of the TRAIL-mediated pathway. HCV NS34A protein appears to be capable of disrupting the innate antiviral host responses in primary human hepatocytes. Our study provides a novel mechanism by which primary human hepatocytes respond to natural HCV infection.


[Complex cases in heart surgery].
Between January 1968 and December 1993, 837 patients underwent cardiac operations with were either complex or performed in the presence of a life threatening disease of other vital organs. Of them 74 (8.8%) patients have died within 30 days after the operation. A substantial number of the operations and associated operative death included left ventricular (LV) aneurysmectomy or plication or LV endoaneurysmectomy with coronary artery bypass (CAB) grafts with or without other cardiac procedures, cardiac reoperations, CAB grafts and mitral or aortic valve replacement, combined mitral and aortic valve replacement (MAVR) with or without tricuspid valve replacement and CAB grafts, CAB grafting for an end-stage coronary artery disease (CAD), emergency CAB grafts for an acute myocardial infarction with cardiogenic shock, complex internal thoracic artery (ITA) grafting, and miscellaneous. The best results were achieved in CAB grafts for an end-stage CAD, complex ITA grafting, CAB grafts with mitral or aortic valve replacement, cardiac reoperations. MAVR and miscellaneous. This is probably, related to an intensive treatment of congestive heart failure (CHF) before the operation, pretreatment with the oxygen free radical inhibitor (allopurinol), selective use of an intraaortic balloon assist device and LV venting, routine use of hemoconcentrator (ultrafiltration) during cardiopulmonary bypass in those with CHF, thorough myocardial protection and a complete coronary revascularization.


Low Energy Availability in Athletes: A Review of Prevalence, Dietary Patterns, Physiological Health, and Sports Performance.
In a high-performance sports environment, athletes can present with low energy availability (LEA) for a variety of reasons, ranging from not consuming enough food for their specific energy requirements to disordered eating behaviors. Both male and female high-performance athletes are at risk of LEA. Longstanding LEA can cause unfavorable physiological and psychological outcomes which have the potential to impair an athlete's health and sports performance. This narrative review summarizes the prevalence of LEA and its associations with athlete health and sports performance. It is evident in the published scientific literature that the methods used to determine LEA and its associated health outcomes vary. This contributes to poor recognition of the condition and its sequelae. This review also identifies interventions designed to improve health outcomes in athletes with LEA and indicates areas which warrant further investigation. While return-to-play guidelines have been developed for healthcare professionals to manage LEA in athletes, behavioral interventions to prevent the condition and manage its associated negative health and performance outcomes are required.


The Peculiar Case of the Hyper-thermostable Pyrimidine Nucleoside Phosphorylase from Thermus thermophilus*.
The poor solubility of many nucleosides and nucleobases in aqueous solution demands harsh reaction conditions (base, heat, cosolvent) in nucleoside phosphorylase-catalyzed processes to facilitate substrate loading beyond the low millimolar range. This, in turn, requires enzymes that can withstand these conditions. Herein, we report that the pyrimidine nucleoside phosphorylase from Thermus thermophilus is active over an exceptionally broad pH (4-10), temperature (up to 100 °C) and cosolvent space (up to 80 % (v/v) nonaqueous medium), and displays tremendous stability under harsh reaction conditions with predicted total turnover numbers of more than 106 for various pyrimidine nucleosides. However, its use as a biocatalyst for preparative applications is critically limited due to its inhibition by nucleobases at low concentrations, which is unprecedented among nonspecific pyrimidine nucleoside phosphorylases.


Finding Novel Antibiotic Substances from Medicinal Plants - Antimicrobial Properties of Nigella Sativa Directed against Multidrug-resistant Bacteria.
The progressive rise in multidrug-resistant (MDR) bacterial strains poses serious problems in the treatment of infectious diseases. While the number of newly developed antimicrobial compounds has greatly fallen, the resistance of pathogens against commonly prescribed drugs is further increasing. This rise in resistance illustrates the need for developing novel therapeutic and preventive antimicrobial options. The medicinal herb Nigella sativa and its derivatives constitute promising candidates. In a comprehensive literature survey (using the PubMed data base), we searched for publications on the antimicrobial effects of N. sativa particularly directed against MDR bacterial strains. In vitro studies published between 2000 and 2015 revealed that N. sativa exerted potent antibacterial effects against both Gram-positive and Gram-negative species including resistant strains. For instance, N. sativa inhibited the growth of bacteria causing significant gastrointestinal morbidity such as Salmonella, Helicobacter pylori, and Escherichia coli. However, Listeria monocytogenes and Pseudomonas aeruginosa displayed resistance against black cumin seed extracts. In conclusion, our literature survey revealed potent antimicrobial properties of N. sativa against MDR strains in vitro that should be further investigated in order to develop novel therapeutic perspectives for combating infectious diseases particularly caused by MDR strains.


Studies on the relationship between estrogen receptor content, glutathione S-transferase pi expression, and induction by 2,3,7,8-tetrachlorodibenzo-p-dioxin and drug resistance in human breast cancer cells.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) induces both phase I and phase II drug-metabolizing enzymes in rodent liver and hepatoma cell lines and this induction is mediated by the aryl hydrocarbon (Ah) receptor. Induction of CYP1A1 by TCDD in human breast cancer cells has been reported and results of several studies suggest that the estrogen receptor (ER) may be required for Ah responsiveness. This study investigates the induction of GST pi by TCDD in human breast cancer cells and the role of the ER in mediating this response. TCDD did not induce chloramphenicol acetyl transferase (CAT) activity in ER positive (ER+) MCF-7 and ER- MDA-MB-468 and MDA-MB-231 human breast cancer cell lines transiently transfected with GST pi (human) or GSTP (rat) promoter-reporter constructs containing the -291/+36 and -2.9/+59 region, respectively, of the GST pi and GSTP gene promoters. Furthermore, TCDD did not induce GST pi or GSTP in MDA-MB-468 and MDA-MB-231 human breast cancer cells stably transfected with the ER. RT-PCR confirmed that GST pi mRNA levels were low in ER+ MCF-7 cells and high in ER- MDA-MB-468 and MDA-MB-231 cells; however, in MDA-MB-468 and MDA-MB-231 cells stably transfected with the ER GST pi mRNA levels remained elevated and were not inducible. MDA-MB-468 and MDA-MB-231 cells stably transfected with the ER exhibited increased GST activity and decreased GSH content compared to wild-type cells; however, in MDA-MB-468 cells stably transfected with ER, the susceptibility to doxorubicin, ellipticine, chlorambucil, malphalan, or cisplatin was similar to that observed in wild-type cells. Adriamycin accumulation was similar in wild-type and ER stably transfected cells and verapamil did not affect this response, suggesting that ER expression did not influence p-glycoprotein activity. Taken together these data suggest that not all GST isoforms are responsive to TCDD and stable transfection of ER- cells with ER is not sufficient to restore the ER+ phenotype in some breast cancer cell lines.


Isotope effects and the nature of enantioselectivity in the shi epoxidation. The importance of asynchronicity.
The epoxidation of beta-methylstyrene catalyzed by the Shi fructose-derived ketone is studied using experimental kinetic isotope effects and DFT calculations. The observation of a large beta olefinic (13)C isotope effect and small alpha carbon isotope effect is indicative of an asynchronous transition state with more advanced formation of the C-O bond to the beta olefinic carbon. By varying the catalyst conformation and alkene orientation, diverse transition structure geometries were located calculationally, and the lowest-energy structure leads to an accurate prediction of the isotope effects. Given this support for the accuracy of the calculations employed, the nature of enantioselectivity in this and related epoxidations is considered. The lowest-energy transition structures are generally those in which the differential formation of the incipient C-O bonds, the "asynchronicity," resembles that of an unhindered model, and the imposition of greater or less asynchronicity leads to higher barriers. In reactions of cis-disubstituted and terminal alkenes using Shi's oxazolidinone catalyst, the asynchronicity of the epoxidation transition state leads to increased steric interaction with the oxazolidinone when a pi-conjugating substituent is distal to the oxazolidinone but decreased steric interaction when the pi-conjugating substituent is proximal to the oxazolidinone. Overall, the asynchronicity of the transition state must be considered carefully to understand the enantioselectivity.


Protocol for the EARCO Registry: a pan-European observational study in patients with α1-antitrypsin deficiency.
Alpha-1 antitrypsin deficiency (AATD) is a genetic condition that leads to an increased risk of emphysema and liver disease. Despite extensive investigation, there remain unanswered questions concerning the natural history, pathophysiology, genetics and the prognosis of the lung disease in association with AATD. The European Alpha-1 Clinical Research Collaboration (EARCO) is designed to bring together researchers from European countries and to create a standardised database for the follow-up of patients with AATD. The EARCO Registry is a non-interventional, multicentre, pan-European, longitudinal observational cohort study enrolling patients with AATD. Data will be collected prospectively without interference/modification of patient's management by the study team. The major inclusion criterion is diagnosed severe AATD, defined by an AAT serum level <11 µM (50 mg·dL-1) and/or a proteinase inhibitor genotype ZZ, SZ or compound heterozygotes or homozygotes of other rare deficient variants. Assessments at baseline and during the yearly follow-up visits include lung function testing (spirometry, body plethysmography and diffusing capacity of the lung), exercise capacity, blood tests and questionnaires (symptoms, quality of life and physical activity). To ensure correct data collection, there will be designated investigator staff to document the data in the case report form. All data will be reviewed by the EARCO database manager. The EARCO Registry aims to understand the natural history and prognosis of AATD better with the goal to create and validate prognostic tools to support medical decision-making.


An interactive image processing system.
A multiuser multiprocessing image processing system has been developed. It is an interactive picture manipulation and enhancement facility which is capable of executing a variety of image processing operations while simultaneously controlling real-time input and output of pictures. It was designed to provide a reliable picture processing system which would be cost-effective in the commercial production environment. Additional goals met by the system include flexibility and ease of operation and modification.


Sonicate Fluid Cellularity Predicted by Transcriptomic Deconvolution Differentiates Infectious from Non-Infectious Arthroplasty Failure.
Although cellularity is traditionally assessed morphologically, deep sequencing approaches being used for microorganism detection may be able to provide information about cellularity. We hypothesized that cellularity predicted using CIBERSORTx (Stanford University), a transcriptomic-based cellular deconvolution tool, would differentiate between infectious and non-infectious arthroplasty failure. CIBERSORTx-derived cellularity profiles of 93 sonicate fluid samples, including 53 from subjects who underwent failed arthroplasties due to periprosthetic joint infection (PJI) (abbreviated for the purpose of this study as PJIF) and 40 from subjects who had undergone non-infectious arthroplasty failure (abbreviated NIAF) that had been subjected to bulk RNA sequencing were evaluated. Samples from PJIF and NIAF subjects were differentially clustered by principal component analysis based on the cellularity profile. Twelve of the 22 individual predicted cellular fractions were differentially expressed in the PJIF cases compared with the NIAF cases, including increased predicted neutrophils (mean and standard error, 9.73% ± 1.06% and 0.81% ± 0.60%), activated mast cells (17.12% ± 1.51% and 4.11% ± 0.44%), and eosinophils (1.96% ± 0.37% and 0.42% ± 0.21%), and decreased predicted M0 macrophages (21.33% ± 1.51% and 39.75% ± 2.45%), M2 macrophages (3.56% ± 0.52% and 8.70% ± 1.08%), and regulatory T cells (1.57% ± 0.23% and 3.20% ± 0.34%). The predicted total granulocyte fraction was elevated in the PJIF cases (32.97% ± 2.13% and 11.76% ± 1.61%), and the samples from the NIAF cases had elevated predicted total macrophage and monocyte (34.71% ± 1.71% and 55.34% ± 2.37%) and total B cell fractions (5.89% ± 0.30% and 8.62% ± 0.86%). Receiver operating characteristic curve analysis identified predicted total granulocytes, neutrophils, and activated mast cells as highly able to differentiate between the PJIF cases and the NIAF cases. Within the PJIF cases, the total granulocyte, total macrophage and monocyte, M0 macrophage, and M2 macrophage fractions were differentially expressed in Staphylococcus aureus compared with Staphylococcus epidermidis -associated samples. Within the NIAF cases, the predicted total B cell, naïve B cell, plasma cell, and M2 macrophage fractions were differentially expressed among different causes of failure. CIBERSORTx can predict the cellularity of sonicate fluid using transcriptomic data, allowing for the evaluation of the underlying immune response during the PJIF and NIAF cases, without a need to phenotypically assess cell composition.


Control of E-cadherin apical localisation and morphogenesis by a SOAP-1/AP-1/clathrin pathway in C. elegans epidermal cells.
E-cadherin (E-cad) is the main component of epithelial junctions in multicellular organisms, where it is essential for cell-cell adhesion. The localisation of E-cad is often strongly polarised in the apico-basal axis. However, the mechanisms required for its polarised distribution are still largely unknown. We performed a systematic RNAi screen in vivo to identify genes required for the strict E-cad apical localisation in C. elegans epithelial epidermal cells. We found that the loss of clathrin, its adaptor AP-1 and the AP-1 interactor SOAP-1 induced a basolateral localisation of E-cad without affecting the apico-basal diffusion barrier. We further found that SOAP-1 controls AP-1 localisation, and that AP-1 is required for clathrin recruitment. Finally, we also show that AP-1 controls E-cad apical delivery and actin organisation during embryonic elongation, the final morphogenetic step of embryogenesis. We therefore propose that a molecular pathway, containing SOAP-1, AP-1 and clathrin, controls the apical delivery of E-cad and morphogenesis.


Intervening Leg Movements Disrupt PLMS Sequences.
Periodic leg movements during sleep (PLMS) occur within a subject as a series with a remarkably stable period defined by the intermovement interval (IMI). Sometimes a non-PLMS movement occurs intervening between two PLMS. PLMS scoring rules totally ignore these intervening leg movements (iLM). This implicitly assumes an iLM results from a process sufficiently independent from the periodic process producing PLMS that it does not affect the periodicity of the surrounding PLMS. This study for the first time tests this basic assumption and explores characteristics of iLM as a potentially significant class of leg movements during sleep. Leg movements were analyzed from two nights of polysomnography recordings from 27 RLS patients and 22 controls using the validated MATPLM1.1 program. All periods (IMI) between PLMS containing an iLM were compared to the local PLMS period defined as the immediately preceding PLMS IMI using pairwise two-sided Wilcoxon sign-rank tests. Similarly, iLM were tested to see if they started a new PLMS series by having the same period as the subsequent PLMS. The periods (IMIs) containing iLM were longer than the previous periods in RLS subjects, but not controls (p < .05). The periods beginning with the iLM were shorter than the subsequent periods in both RLS and controls (p < .05). iLM as a separate type of LM distort PLMS periodicity and do not restart PLMS series. iLM end PLMS series.


Fatal toxicity associated with antidepressant use in primary care.
New selective serotonin reuptake inhibitors (SSRIs) are perceived to be much safer in use than older tricyclic antidepressants (TCAs). However, previous assessments of association with fatal toxicity were made too soon after the introduction of the new drugs to permit accurate estimation. To determine the level of association of antidepressant drugs with fatal poisoning in the treatment of depression. National data for England and Wales for three years (1993 to 1995) for fatal poisonings associated with antidepressants were obtained and, together with national primary care data on prescribing, were used to calculate fatality association by antidepressant drug. There were substantial variations between drugs in the level of association with fatal poisoning. Assuming an average treatment episode lasted three months, one fatality is associated with 11,800 treatment episodes of antidepressant use (95% CI = 11,120 to 12,580) when only single substance fatalities are considered. For SSRIs as a group the association was one in 411,800 (95% CI = 243,300 to 1.34 million) and for TCAs one in 8130 (95% CI = 7650 to 8670). However, for one of the newer TCAs, lofepramine, the single substance fatality rate associated with its use was one in 233,700 (95% CI = 124,500 to 1.89 million), which is not statistically significantly different from the SSRIs (P = 0.35). Estimated death rates associated with specific antidepressants should be compared with caution because drugs may be used selectively in patients with differing severity of depression. The proportion of these fatalities that could be prevented by switching to safer antidepressants is unclear when so few deaths are recorded as accidental; when there is intent to do self-harm the potential for switching to other means is unknown. However, this approach to relative toxicity may remain the best available since it is unlikely that a randomised trial will ever be conducted with a large enough sample size to obtain experimental data. Fatalities from antidepressant poisoning are very rare but if safety is paramount then lofepramine or an SSRI are justifiable treatment choices.


Risk Factors Associated With Glaucomatous Progression in Pseudoexfoliation Patients.
To investigate risk factors associated with glaucomatous progression in pseudoexfoliation glaucoma (PXG) patients. A retrospective observational study with a total of 48 PXG patients and age, sex, and visual field (VF) test mean deviation-matched 48 primary open-angle glaucoma (POAG) patients recruited from 3 university-based hospitals. Glaucomatous progression was defined by 2 criteria, either optic disc/retinal nerve fiber layer photographic assessment or serial VF data. Univariate and multivariate Cox proportional hazard models were used to detect associations between risk factors and progression in each subgroup and shared frailty model were used in total participants. In total participants, PXG diagnosis [hazard ratio (HR), 7.245; P=0.050], a higher baseline intraocular pressure (IOP) (HR, 1.221; P=0.025) and a greater number of glaucoma medications (HR, 3.537; P=0.016) were significant factors for glaucoma progression. In the PXG group, a higher baseline IOP (HR, 1.131; P<0.001), a lower IOP reduction (HR, 0.966; P=0.027), a greater number of glaucoma medications (HR, 2.413; P=0.004), and worse baseline VF mean deviation (HR, 0.875; P=0.004) were significant factors, whereas the number of glaucoma medications (HR, 3.493; P=0.019) and disc hemorrhage (HR, 3.953; P=0.043) showed association in the POAG group. Factors associated with progression differed between PXG and POAG groups. Higher baseline IOP and requirement for a greater number of glaucoma medications were related with PXG progression.


The effect of aspirin and eicosapentaenoic acid on urinary biomarkers of prostaglandin E2 synthesis and platelet activation in participants of the seAFOod polyp prevention trial.
Urinary prostaglandin (PG) E metabolite (PGE-M) and 11-dehydro (d)-thromboxane (TX) B2 are biomarkers of cyclooxygenase-dependent prostanoid synthesis. We investigated (1) the effect of aspirin 300 mg daily and eicosapentaenoic acid (EPA) 2000 mg daily, alone and in combination, on urinary biomarker levels and, (2) whether urinary biomarker levels predicted colorectal polyp risk, during participation in the seAFOod polyp prevention trial. Urinary PGE-M and 11-d-TXB2 were measured by liquid chromatography-tandem mass spectrometry. The relationship between urinary biomarker levels and colorectal polyp outcomes was investigated using negative binomial (polyp number) and logistic (% with one or more polyps) regression models. Despite wide temporal variability in PGE-M and 11-d-TXB2 levels within individuals, both aspirin and, to a lesser extent, EPA decreased levels of both biomarkers (74% [P ≤ .001] and 8% [P ≤ .05] reduction in median 11-d-TXB2 values, respectively). In the placebo group, a high (quartile [Q] 2-4) baseline 11-d-TXB2 level predicted increased polyp number (incidence rate ratio [IRR] [95% CI] 2.26 [1.11,4.58]) and risk (odds ratio [95% CI] 3.56 [1.09,11.63]). A low (Q1) on-treatment 11-d-TXB2 level predicted reduced colorectal polyp number compared to placebo (IRR 0.34 [0.12,0.93] for combination aspirin and EPA treatment) compared to high on-treatment 11-d-TXB2 values (0.61 [0.34,1.11]). Aspirin and EPA both inhibit PGE-M and 11-d-TXB2 synthesis in keeping with shared in vivo cyclooxygenase inhibition. Colorectal polyp risk and treatment response prediction by 11-d-TXB2 is consistent with a role for platelet activation during early colorectal carcinogenesis. The use of urinary 11-d-TXB2 measurement for a precision approach to colorectal cancer risk prediction and chemoprevention requires prospective evaluation.


Diversification of freshwater crabs on the sky islands in the Hengduan Mountains Region, China.
The numerous naturally-fragmented sky islands (SIs) in the Hengduan Mountains Region (HMR) of southwestern China constitute discontinuous landscapes where montane habitats are isolated by dry-hot valleys which have fostered exceptional species diversification and endemicity. However, studies documenting the crucial role of SI on the speciation dynamics of native freshwater organisms are scarce. Here we used a novel set of comprehensive genetic markers (24 nuclear DNA sequences and complete mitogenomes), morphological characters, and biogeographical information to reveal the evolutionary history and speciation mechanisms of a group of small-bodied montane potamids in the genus Tenuipotamon. Our results provide a robustly supported phylogeny, and suggest that the vicariance events of these montane crabs correlate well with the emergence of SIs due to the uplift of the HMR during the Late Oligocene. Furthermore, ancestrally, mountain ridges provided corridors for the dispersal of these montane crabs that led to the colonization of moist montane-specific habitats, aided by past climatic conditions that were the crucial determinants of their evolutionary history. The present results illustrated that the mechanisms isolating SIs are reinforced by the harsh-dry isolating climatic features of dry-hot valleys separating SIs and continue to affect local diversification. This offers insights into the causes of the high biodiversity and endemism shown by the freshwater crabs of the HMR-SIs in southwestern China.


Regiospecific functionalization of methyl C-H bonds of alkyl groups in reagents with heteroatom functionality.
We report the regiospecific, rhodium-catalyzed borylation of saturated terminal C-H bonds in molecules with existing functionality. Moderate to good yields were obtained with the organic substrate in excess and as limiting reagent. The borylations of trialkylamines, protected alcohols, protected ketones, and fluoroalkanes occurred regiospecifically at the methyl group that is least sterically hindered. Reactions were also conducted that probed electronic effects on the alkyl borylation. These reactions showed that the borylation occurred preferentially at the methyl group that is most electron-deficient. Methods to conduct tandem borylation of C-H bonds and conversion of the resulting boronate esters to alcohols, alkylarenes, and alkyltrifluoroborates were also developed.


MELOGEN: an EST database for melon functional genomics.
Melon (Cucumis melo L.) is one of the most important fleshy fruits for fresh consumption. Despite this, few genomic resources exist for this species. To facilitate the discovery of genes involved in essential traits, such as fruit development, fruit maturation and disease resistance, and to speed up the process of breeding new and better adapted melon varieties, we have produced a large collection of expressed sequence tags (ESTs) from eight normalized cDNA libraries from different tissues in different physiological conditions. We determined over 30,000 ESTs that were clustered into 16,637 non-redundant sequences or unigenes, comprising 6,023 tentative consensus sequences (contigs) and 10,614 unclustered sequences (singletons). Many potential molecular markers were identified in the melon dataset: 1,052 potential simple sequence repeats (SSRs) and 356 single nucleotide polymorphisms (SNPs) were found. Sixty-nine percent of the melon unigenes showed a significant similarity with proteins in databases. Functional classification of the unigenes was carried out following the Gene Ontology scheme. In total, 9,402 unigenes were mapped to one or more ontology. Remarkably, the distributions of melon and Arabidopsis unigenes followed similar tendencies, suggesting that the melon dataset is representative of the whole melon transcriptome. Bioinformatic analyses primarily focused on potential precursors of melon micro RNAs (miRNAs) in the melon dataset, but many other genes potentially controlling disease resistance and fruit quality traits were also identified. Patterns of transcript accumulation were characterised by Real-Time-qPCR for 20 of these genes. The collection of ESTs characterised here represents a substantial increase on the genetic information available for melon. A database (MELOGEN) which contains all EST sequences, contig images and several tools for analysis and data mining has been created. This set of sequences constitutes also the basis for an oligo-based microarray for melon that is being used in experiments to further analyse the melon transcriptome.


Efficacy of combined topical treatment of acne vulgaris with adapalene and nadifloxacin: a randomized study.
Topical retinoid and antibiotic combination therapy is an integral part of acne treatment and is considered the appropriate first-line therapy according to the Japanese guideline for moderate and severe acne. In this combination, clindamycin or doxycycline are mostly used as antibiotics, but there have been no reports on the effectiveness of nadifloxacin, a widely used antibiotic in Japan and European countries for acne, in combination with topical retinoid. To confirm the efficacy and safety of adapalene gel and nadifloxacin cream in the treatment of Japanese patients with acne vulgaris, a total of 50 patients were randomized to the two groups, the combination therapy and the adapalene monotherapy, and each therapy was tested for 8 weeks. The percentage reduction in the number of inflammatory acne lesions was evaluated and the safety was monitored through adverse events. The combination of adapalene gel and nadifloxacin cream produced a significantly higher reduction in the inflammatory lesions at 2 weeks (P = 0.047) and at 8 weeks (P = 0.011) after the starting than did adapalene gel monotherapy. The combination did not elevate the side effects of erythema and scale scores, but rather significantly depressed erythema at 1 week. This study showed the efficacy and safety of the combination therapy of nadifloxacin cream with adapalene gel for the inflammatory acne.


A novel 2-bp indel within Krüppel-like factor 15 gene (KLF15) and its associations with chicken growth and carcass traits.
1. The Krüppel-like factor (KLF) family of zinc-finger transcription factors plays a critical role in cell differentiation, phenotypic modulation and physiologic function. KLF15 has been proposed to regulate adipogenesis and gluconeogenesis. The objective of this study was to establish the association between KLF15 gene polymorphism and chicken growth and carcass traits. 2. An F2resource population of Gushi chickens crossed with Anka broilers was used to investigate the genetic effects of the chicken KLF15 gene. A 2-bp indel mutation (G13781_13782del/insAG) within intron 2 was detected, and a polymerase chain reaction-restriction fragment length polymorphism method was developed to genotype the F2 individuals. 3. Association analysis showed that the Single Nucleotide Polymorphisms (SNP) was significantly associated with chicken growth and carcass traits. The chickens with the insAG/insAG genotype generally had a significantly higher body weight and size than other genotypes. Gene expression for each genotype showed that birds carrying insAG/insAG had a higher expression level than the other genotypes. 4. The results suggested that this polymorphic site may serve as a useful target for marker assisted selection of chicken growth and carcass traits.


A cluster-randomized trial to reduce major perinatal morbidity among women with one prior cesarean delivery in Québec (PRISMA trial): study protocol for a randomized controlled trial.
Rates of cesarean delivery are continuously increasing in industrialized countries, with repeated cesarean accounting for about a third of all cesareans. Women who have undergone a first cesarean are facing a difficult choice for their next pregnancy, i.e.: (1) to plan for a second cesarean delivery, associated with higher risk of maternal complications than vaginal delivery; or (b) to have a trial of labor (TOL) with the aim to achieve a vaginal birth after cesarean (VBAC) and to accept a significant, but rare, risk of uterine rupture and its related maternal and neonatal complications. The objective of this trial is to assess whether a multifaceted intervention would reduce the rate of major perinatal morbidity among women with one prior cesarean. The study is a stratified, non-blinded, cluster-randomized, parallel-group trial of a multifaceted intervention. Hospitals in Quebec are the units of randomization and women are the units of analysis. As depicted in Figure 1, the study includes a 1-year pre-intervention period (baseline), a 5-month implementation period, and a 2-year intervention period. At the end of the baseline period, 20 hospitals will be allocated to the intervention group and 20 to the control group, using a randomization stratified by level of care. Medical records will be used to collect data before and during the intervention period. Primary outcome is the rate of a composite of major perinatal morbidities measured during the intervention period. Secondary outcomes include major and minor maternal morbidity; minor perinatal morbidity; and TOL and VBAC rate. The effect of the intervention will be assessed using the multivariable generalized-estimating-equations extension of logistic regression. The evaluation will include subgroup analyses for preterm and term birth, and a cost-effectiveness analysis. The intervention is designed to facilitate: (1) women's decision-making process, using a decision analysis tool (DAT), (2) an estimate of uterine rupture risk during TOL using ultrasound evaluation of low-uterine segment thickness, (3) an estimate of chance of TOL success, using a validated prediction tool, and (4) the implementation of best practices for intrapartum management. Current Controlled Trials, ID: ISRCTN15346559 . Registered on 20 August 2015.


Benchmarking the scientific output of industrial wastewater research in Arab world by utilizing bibliometric techniques.
Rapid population growth, worsening of the climate, and severity of freshwater scarcity are global challenges. In Arab world countries, where water resources are becoming increasingly scarce, the recycling of industrial wastewater could improve the efficiency of freshwater use. The benchmarking of scientific output of industrial wastewater research in the Arab world is an initiative that could support in shaping up and improving future research activities. This study assesses the scientific output of industrial wastewater research in the Arab world. A total of 2032 documents related to industrial wastewater were retrieved from 152 journals indexed in the Scopus databases; this represents 3.6 % of the global research output. The h-index of the retrieved documents was 70. The total number of citations, at the time of data analysis, was 34,296 with an average citation of 16.88 per document. Egypt, with a total publications of 655 (32.2 %), was ranked the first among the Arab countries followed by Saudi Arabia 300 (14.7 %) and Tunisia 297 (14.6 %). Egypt also had the highest h-index, assumed with Saudi Arabia, the first place in collaboration with other countries. Seven hundred fifteen (35.2 %) documents with 66 countries in Arab/non-Arab country collaborations were identified. Arab researchers collaborated mostly with researchers from France 239 (11.7 %), followed by the USA 127 (6.2 %). The top active journal was Desalination 126 (6.2 %), and the most productive institution was the National Research Center, Egypt 169 (8.3 %), followed by the King Abdul-Aziz University, Saudi Arabia 75 (3.7 %). Environmental Science was the most prevalent field of interest 930 (45.8 %). Despite the promising indicators, there is a need to close the gap in research between the Arab world and the other nations. Optimizing the investments and developing regional experiences are key factors to promote the scientific research.


Climate warming and Bergmann's rule through time: is there any evidence?
Climate change is expected to induce many ecological and evolutionary changes. Among these is the hypothesis that climate warming will cause a reduction in body size. This hypothesis stems from Bergmann's rule, a trend whereby species exhibit a smaller body size in warmer climates, and larger body size under colder conditions in endotherms. The mechanisms behind this rule are still debated, and it is not clear whether Bergmann's rule can be extended to predict the effects of climate change through time. We reviewed the primary literature for evidence (i) of a decrease in body size in response to climate warming, (ii) that changing body size is an adaptive response and (iii) that these responses are evolutionary or plastic. We found weak evidence for changes in body size through time as predicted by Bergmann's rule. Only three studies investigated the adaptive nature of these size decreases. Of these, none reported evidence of selection for smaller size or of a genetic basis for the size change, suggesting that size decreases could be due to nonadaptive plasticity in response to changing environmental conditions. More studies are needed before firm conclusions can be drawn about the underlying causes of these changes in body size in response to a warming climate.


Fostering Transformative Learning in a Social Pediatrics Research Summer Studentship.
Teaching future doctors the skills necessary to address health disparities is a challenge for medical educators. In response, the authors developed and implemented the Social Pediatrics Research Summer Studentship (SPReSS) program for medical students at the University of Toronto. The curriculum incorporated research and clinical placements into a formal seminar series. Participating students were required to complete a research project and to write a reflection describing a situation that challenged their thinking. The authors and curriculum developers applied transformative learning principles not only to facilitate critical reflection and learning in the students but also as an innovative approach to program development and evaluation. The authors conducted a thematic analysis of the reflections of 23 students participating in the program in June and July 2013, 2014, and 2015 to evaluate the SPReSS program. The analysis revealed students' empathic responses to marginalized patients, and these responses acted as triggers for critical reflection. Students described feeling empowered to act as advocates and wrote that these feelings were reinforced through faculty members' role modeling. According to their reflections, students found the program both challenging and rewarding, particularly the integration of the clinical and research experiences which made broader sociopolitical phenomena introduced through assigned readings and seminar discussions concrete. The authors are exploring models, including a fourth-year selective or multiyear longitudinal experience, to support more students. They also hope to involve more community partners and to evaluate long-term outcomes of participants.


Mechanisms of postabsorptive hyperglycemia in streptozotocin diabetic rats.
Postabsorptive hepatic glucose output (HGO) was estimated in normal (n = 9) and streptozotocin (STZ) diabetic rats after a 6-h [3-3H]glucose infusion. In diabetic rats, HGO was estimated at ambient (n = 12) or normal (achieved via phlorizin infusion; n = 9) glucose concentrations. HGO was not statistically different between normal and diabetic rats (63 +/- 3 vs. 77 +/- 10 mumol.kg-1.min-1; P > 0.05). HGO was also normal in diabetic rats even when plasma glucose was normalized with phlorizin infusion (71 +/- 5 vs. 63 +/- 3 mumol.kg-1.min-1; P > 0.05). In contrast, peripheral glucose uptake, when estimated at matched euglycemia, was lower by approximately 25% in diabetic than in normal rate (46 +/- 6 vs. 62 +/- 3 mumol.kg-1.min-1; P < 0.01). In addition, acute changes in plasma glucose concentrations did not have significant effects on HGO or peripheral glucose uptake in diabetic rats (P > 0.05), resulting in markedly decreased glucose clearance at ambient hyperglycemia (P < 0.001). In conclusion, postabsorptive HGO was not elevated in a majority (17 of 21) of STZ diabetic rats with severe hyperglycemia and therefore was not responsible for postabsorptive hyperglycemia. Our data suggest that an impairment in the ability of glucose to regulate peripheral glucose uptake or HGO develops in STZ diabetes and contributes to postabsorptive hyperglycemia.


Serotonin-induced protein phosphorylation in a lobster neuromuscular preparation.
Three substances believed to be neurohormones in the lobster--serotonin, octopamine, and proctolin--can modulate the physiology of the dactyl opener muscle of the lobster walking leg. All three act directly on muscle fibers to cause long-lasting contractures. In addition, serotonin enhances the release of transmitter from the excitatory and inhibitory axons that innervate the muscle. We now report that serotonin, applied to intact nerve-muscle preparations, increases the phosphorylation of a soluble 29,000-dalton protein by as much as 20-fold. In two respects the phosphorylation parallels the physiological effects of serotonin: both have a slow time course, and both are caused by low concentrations of serotonin (thresholds of approximately 10(-8) M). The 29,000-dalton protein is present in many lobster tissues, including thoracic ganglia, gills, and several different muscles. This suggests that the phosphorylation participates in some aspect of metabolism common to many types of cells, rather than in an activity unique to nerve or muscle. The function of the phosphorylated protein remains unknown. Several lines of evidence suggest that the phosphorylation is mediated by a cyclic nucleotide, probably cyclic AMP. (1) Adding cyclic AMP or cyclic GMP to nerve-muscle homogenates leads to the phosphorylation. (2) Adding 8-Br-cyclic AMP or 8-Br-cyclic GMP to intact preparations leads to the phosphorylation. (3) In intact tissue, forskolin, an adenylate cyclase activator, and isobutylmethylxanthine, a phosphodiesterase inhibitor, increase cyclic AMP and cyclic GMP and also cause the phosphorylation. (4) Serotonin increases cyclic AMP in intact preparations. (5) RMI 12,330A, which blocks the serotonin-induced cyclic AMP increase, also blocks the phosphorylation. (6) Serotonin has no detectable effect on cyclic GMP. This suggests that cyclic AMP, and not cyclic GMP, is the second messenger that mediates phosphorylation in vivo. In contrast, calcium is not likely to mediate the phosphorylation. Agents that block calcium channels do not block the serotonin-induced phosphorylation, and agents that increase sarcoplasmic calcium do not cause it. Furthermore, calcium does not induce the phosphorylation in a homogenate. One of the other modulators, proctolin, does not increase cyclic AMP or cause the phosphorylation even at concentrations 10(6)-fold greater than the hormone's threshold for producing contractures. The third modulator, octopamine, causes small increases in cyclic AMP and is a weak agonist of the phosphorylation. Neither proctolin nor octopamine alters cyclic GMP levels.


[Contamination, by Toxocara spp eggs, in public parks and squares in Botucatu, São Paulo, Brazil].
The frequency of Toxocara spp eggs in public parks was determined from March 1995 to February 1996. One hundred and twelve samples were collected from 10 public parks in Botucatu, São Paulo State, Brazil. Samples were processed by the decinormal sodium hydroxide concentration method. Out of the 120 soil samples analyzed, 21 were contaminated with Toxocara spp eggs, corresponding to a 17.5% rate of infestation. Of the ten squares submitted to analysis during the year, six presented contamination. Most of the ova found presented characteristics of inviability, with a consequent low chance of human infection, although the population is not free from the risk of contracting this zoonosis.


Have incidence rates of liver cancer peaked in the United States?
Liver cancer incidence has increased for several decades in the United States. Recently, reports have suggested that rates of hepatocellular carcinoma (HCC), the dominant form of liver cancer, had declined in certain groups. However, to the authors' knowledge, the most recent histology-specific liver cancer rates have not been reported to date. The authors examined the incidence of HCC and intrahepatic cholangiocarcinoma (ICC) from 1992 through 2016 using data from the Surveillance, Epidemiology, and End Results registries. Age-standardized incidence rates were calculated by histology, sex, race and/or ethnicity, and age. Trends were analyzed using the National Cancer Institute's Joinpoint Regression Program to estimate the annual percent change. Between 2011 and 2016, HCC rates significantly declined (annual percent change, -1.9%), with more prominent declines noted among males, Asian/Pacific Islanders, and individuals aged <50 years. Conversely, ICC rates increased from 2002 through 2016. Declining HCC rates may persist due to improved treatment of the hepatitis C virus and/or competing causes of mortality among individuals with fatty liver disease.


Effect of hemodialysis and peritoneal dialysis on redox status in chronic renal failure patients: a comparative study.
To investigate the effects of hemodialysis (HD) and periotoneal dialysis (PD) on oxidative stress in chronic renal failure patients (CRF). 20 HD patients (M/F: 8/12, 36 ± 12 years) and 20 PD patients (M/F: 10/10, 40 ± 8 years) were compared with 20 end stage renal failure patients (CRF) (M/F: 4/16, 61 ± 13 years). Thiobarbituric acid reactive substances (TBARS) values were elevated in HD and decreased in PD compared to CRF (P < 0.05). TBARS-VLDL and TBARS-HDL2 were decreased in HD and PD, compared to CRF (p < 0.05). TBARS-LDL were higher in HD compared to CRF (p < 0.05). No significant difference in TBARS-HDL3 values between the three groups. Carbonyls were increased in HD (p < 0.05) and PD (p < 0.01) compared to CRF. Plasma superoxide dismutase activity (SOD) was decreased in HD compared to CRF and PD (P < 0.05). Glutathion peroxidase activity (GSH-Px) was decreased in HD and PD (P < 0.005), compared to CRF. Decrease in catalase activity was noted only in PD compared to CRF (P < 0.05). An increase in nitric oxide was noted in HD compared to CRF (p < 0.05). Albumin concentrations were higher in HD and PD compared to CRF (P < 0.001). Whereas uric acid concentrations were decreased in HD (P < 0.001) compared to CRF and PD. Bilirubin values were similar in all groups. Increased values of iron were noted in HD and PD, compared to PD (p < 0.001). HD and PD aggravate oxidative stress generated by uremia. HD accentuates lipid and protein peroxidation, while PD aggravates protein oxidation. However, the activity of antioxidant enzymes was altered by both dialysis treatments.


[The "phenomenon of ethanol preference"].
The problem of using the "phenomenon of ethanol preference" under conditions of a free choice between ethanol and water as a behavioural model of alcoholism is discussed. The methodical aspects of the model are considered: the regimen of the animals' keeping, the criteria of preference, the concentrations of ethanol solutions, the schedule of presentation of the solutions. There was shown the principal similarity between the changes in the behaviour and biological parameters of the animals preferring ethanol and the changes observed in the man suffering from chronic alcoholism. It is concluded that despite the imperfection of the model the "phenomenon of ethanol preference" remains fairly valuable for studying the etiopathogenesis of alcoholism and developing the methods of the therapeutic action on the patient.


The Na(+)/Ca(2+), K(+) exchanger 2 modulates mammalian cone phototransduction.
Calcium ions (Ca(2+)) modulate the phototransduction cascade of vertebrate cone photoreceptors to tune gain, inactivation, and light adaptation. In darkness, the continuous current entering the cone outer segment through cGMP-gated (CNG) channels is carried in part by Ca(2+), which is then extruded back to the extracellular space. The mechanism of Ca(2+) extrusion from mammalian cones is not understood. The dominant view has been that the cone-specific isoform of the Na(+)/Ca(2+), K(+) exchanger, NCKX2, is responsible for removing Ca(2+) from their outer segments. However, indirect evaluation of cone function in NCKX2-deficient (Nckx2(-/-)) mice by electroretinogram recordings revealed normal photopic b-wave responses. This unexpected result suggested that NCKX2 may not be involved in the Ca(2+) homeostasis of mammalian cones. To address this controversy, we examined the expression of NCKX2 in mouse cones and performed transretinal recordings from Nckx2(-/-) mice to determine the effect of NCKX2 deletion on cone function directly. We found that Nckx2(-/-) cones exhibit compromised phototransduction inactivation, slower response recovery and delayed background adaptation. We conclude that NCKX2 is required for the maintenance of efficient Ca(2+) extrusion from mouse cones. However, surprisingly, Nckx2(-/-) cones adapted normally in steady background light, indicating the existence of additional Ca(2+)-extruding mechanisms in mammalian cones.


Australia and New Zealand Islets and Pancreas Transplant Registry Annual Report 2017-Pancreas Waiting List, Recipients, and Donors.
This is a registry report from the Australia and New Zealand Islet and Pancreas Transplant Registry. We report data for all solid organ pancreas transplant activity from inception in 1984 to end of 2016. Data analysis was performed using Stata Software version 14 (StataCorp, College Station, Tex). From 1984 to 2016 a total of 756 solid organ pancreas transplants have been performed in Australia and New Zealand, in 738 individuals. In 2016, 55 people received a pancreas transplant. These transplants were performed in Auckland (4), Monash (22), and Westmead (29). In 2016, 50 transplants were simultaneous pancreas kidney, 4 were pancreas after kidney, and 1 was a pancreas transplant alone.


Normal gut microbiome in NMDA receptor encephalitis.
To determine whether the gut microbiota shows overabundance of commensal bacteria species in patients with anti-NMDA receptor (NMDAR) encephalitis, similar to patients with MS or neuromyelitis optica where they potentially balance pro- and anti-inflammatory immune responses or participate in disease pathogenesis by molecular mimicry. Intestinal microbiota was characterized in patients with NMDAR encephalitis (n = 23, mean age: 34 ± 12.7 years; 21 females) and age/sex/environment-matched healthy controls (n = 24, 40 ± 14.2 years; 22 females) using stool bacteria 16S rDNA sequencing and classification in operational taxonomic units (OTUs). Statistical analyses focused on intraindividual and interindividual bacterial diversity and identification of differentially abundant taxa. Patients with NMDAR encephalitis and controls had similar microbiome profiles of the gut microbiota regarding intraindividual bacterial diversity, OTU distribution, ratio between regional and local species diversity when testing all OTUs, and genera with a relative abundance greater than 0.5%. Similarly, the subgroup of NMDAR encephalitis patients with an ovarian teratoma (n = 3) showed no differences in microbiome variation compared with controls. Patients in the acute encephalitis stage (n = 8) showed significant differences in the numbers of Clostridium XVIII, Clostridium IV, Oscillibacter, Prevotella, and Blautia; however, significance was lost after correction for multiple testing. Patients with NMDAR encephalitis and controls both had a normal gut microbiome. The lack of overabundance of certain bacterial species in patients suggests that microbiome changes are no major contributors to the pathogenesis, disease course, or prognosis in NMDAR encephalitis. Despite the small sample size and heterogeneous groups, findings indicate differences to other neuroimmunologic diseases.


Generation of human acute lymphoblastic leukemia xenografts for use in oncology drug discovery.
The establishment of reproducible mouse models of acute lymphoblastic leukemia (ALL) is necessary to provide in vivo therapeutic test systems that recapitulate human ALL, and for amplification of limited amounts of primary tumor material. A popular assay is the primary xenograft model that utilizes immunocompromised mice. The protocol includes injection of primary patient tumor specimens into mice with subsequent serial passaging of the tumors by retransplants of cells harvested from the mouse bone marrow and spleen. The tumors generated are then used for genomic profiling, ex vivo compound testing, mechanistic studies and retransplantation. Detailed in this unit are procedures for the establishment and maintenance of primary ALL xenograft panels for use in basic research and translational studies.


Dynamic regulation of p53 subnuclear localization and senescence by MORC3.
The tumor suppressor p53 is a key transcriptional factor regulating the induction of cellular senescence by oncogenic signals. The activity of p53 is regulated by recruitment into promyelocytic leukemia (PML)-nuclear bodies (NBs) as well as by stabilization through posttranslational modifications such as phosphorylation and acetylation. Here we found that MORC3 (microrchidia3)-ATPase activated p53 and induced cellular senescence in normal human and mouse fibroblasts but not p53-/- fibroblasts. Conversely, genotoxic stress-induced phosphorylation and stabilization of p53 but barely increased its transcriptional activity in Morc3-/- fibroblasts. MORC3 localized on PML-NBs in presence of PML and mediated recruitment of p53 and CREB-binding protein (CBP) into PML-NBs. In contrast, expression of ATPase activity-deficient mutant MORC3-E35A or siRNA repression of MORC3 impaired the localization of p53 and Sp100 but not CBP on PML-NBs. These results suggest that MORC3 regulates p53 activity and localization into PML-NBs. We identified a new molecular mechanism that regulates the activity of nuclear proteins by localization to a nuclear subdomain.


Multilead ST-segment monitoring in patients with acute coronary syndromes: a consensus statement for healthcare professionals. ST- Segment Monitoring Practice Guideline International Working Group.
ST-segment monitoring is underused by healthcare professionals for patients with acute coronary syndromes treated in emergency departments and intensive care units. To provide clinically practical consensus guidelines for optimal ST-segment monitoring. A working group of key nurses and physicians met in Dallas, Tex, in November 1998. Consensus was reached on who should and should not have ST monitoring, goals and time frames for ST monitoring in various diagnostic categories, what electrocardiographic leads should be monitored, what equipment requirements are needed, what strategies improve accuracy and clinical usefulness of ST monitoring, and what knowledge and skills are required for safe and effective ST monitoring. Because changes in the ST segment can shift among various electrocardiographic leads in the same person over time owing to different ischemic mechanisms, 12-lead ST monitoring is recommended. Recommended monitoring times are as follows: myocardial infarction or unstable angina, 24 to 48 hours or until patient is event-free for 12 to 24 hours; chest pain prompting a visit to an emergency department, 8 to 12 hours; catheter-based interventions with less definitive interventional outcomes requiring monitoring in an intensive unit, 6 to 12 hours; and cardiac surgery or noncardiac surgery in patients with coronary disease or risk factors, 24 to 48 hours. An ST measurement point of J + 60 ms makes it unlikely that measurement will coincide with the upslope of the T wave, even in patients with sinus tachycardia. Accurate and consistent lead placement and careful electrode and skin preparation are imperative to improve the clinical usefulness of ST monitoring.


Synthetic associative learning in engineered multicellular consortia.
Associative learning (AL) is one of the key mechanisms displayed by living organisms in order to adapt to their changing environments. It was recognized early as a general trait of complex multicellular organisms but is also found in 'simpler' ones. It has also been explored within synthetic biology using molecular circuits that are directly inspired in neural network models of conditioning. These designs involve complex wiring diagrams to be implemented within one single cell, and the presence of diverse molecular wires become a challenge that might be very difficult to overcome. Here we present three alternative circuit designs based on two-cell microbial consortia able to properly display AL responses to two classes of stimuli and displaying long- and short-term memory (i.e. the association can be lost with time). These designs might be a helpful approach for engineering the human gut microbiome or even synthetic organoids, defining a new class of decision-making biological circuits capable of memory and adaptation to changing conditions. The potential implications and extensions are outlined.


Use of MRI to identify enlarged inferior gluteal and ischioanal lymph nodes and associated findings related to the primary disease.
We aimed to draw attention to the lymph nodes at the inferior gluteal and ischioanal regions and evaluate the lesions accompanying them using 3.0 T magnetic resonance imaging (MRI). In total, 22 patients (15 men, 7 women; mean age, 50±11.2 years; age range, 32-71 years) were included in this study. The patients' medical records were reviewed. MRI data were reviewed on a picture archiving and communication system workstation by two radiologists in consensus. Lymph node location, laterality, number, and size were documented. The primary disorders causing the enlargement of inferior gluteal lymph nodes (n=16) were perianal fistula of cryptoglandular origin (n=5), perianal fistula associated with Crohn's disease (n=2), decubitus ulcers (n=2), presacral abscess (n=1), non-Hodgkin lymphoma (n=2), prostate cancer invading urethra and anorectal junction (n=1), endometrium cancer invading the urethra and vagina (n=1), and anal cancer (n=2). The pathologies causing the enlargement of ischioanal lymph nodes (n=6) were perianal fistula of cryptoglandular origin (n=4), subcutaneous inflammation of gluteal region related to Crohn's disease (n=1), and prostate cancer (n=1). The infectious and neoplastic lesions involving the anal canal, distal rectum, gluteal region, prostate, and urethra are the possible causes of inferior gluteal and ischioanal lymph node enlargement. Lymphoproliferative diseases can also affect these node groups. MRI is an important method to identify enlarged inferior gluteal and ischioanal lymph nodes and define associated findings.


Loss of chondroitin 6-O-sulfotransferase-1 function results in severe human chondrodysplasia with progressive spinal involvement.
We studied two large consanguineous families from Oman with a distinct form of spondyloepiphyseal dysplasia (SED Omani type). By using a genome-wide linkage approach, we were able to map the underlying gene to a 4.5-centimorgan interval on chromosome 10q23. We sequenced candidate genes from the region and identified a missense mutation in the chondroitin 6-O-sulfotransferase (C6ST-1) gene (CHST3) changing an arginine into a glutamine (R304Q) in the well conserved 3'-phosphoadenosine 5'-phosphosulfate binding site. C6ST-1 catalyzes the modifying step of chondroitin sulfate (CS) synthesis by transferring sulfate to the C-6 position of the N-acetylgalactosamine of chondroitin. From the crystal structures of other sulfotransferases, it could be inferred that Arg-304 is essential for the structure of the cosubstrate binding site. We used recombinant C6ST-1 to show that the identified missense mutation completely abolishes C6ST-1 activity. Disaccharide composition analysis of CS chains by anion-exchange HPLC shows that both Delta HexA-GalNAc(6S) and Delta HexA(2S)-GalNAc(6S) were significantly reduced in the patient's cells and that Delta HexA-GalNAc(4S,6S), undetectable in controls, was elevated. Analysis of the patient's urine shows marked undersulfation of CS, in particular reduction in 6-O-sulfated disaccharide and an increase in the nonsulfated unit. Our results indicate that the mutation in CHST3 described here causes a specific but generalized defect of CS chain sulfation resulting in chondrodysplasia with major involvement of the spine.


Growth inhibition and radiosensitization of glioblastoma and lung cancer cells by small interfering RNA silencing of tumor necrosis factor receptor-associated factor 2.
Radiotherapy combined with chemotherapy is the treatment of choice for glioblastoma and locally advanced lung cancer, but radioresistance of these two types of cancer remains a significant therapeutic hindrance. To identify molecular target(s) for radiosensitization, we screened a small interfering RNA (siRNA) library targeting all protein kinases and E3 ubiquitin ligases in the human genome and identified tumor necrosis factor receptor-associated factor 2 (TRAF2). Silencing of TRAF2 using siRNA caused a significant growth suppression of glioblastoma U251 cells and moderately sensitized these radioresistant cells to radiation. Overexpression of a really interesting new gene (RING)-deleted dominant-negative TRAF2 mutant also conferred radiosensitivity, whereas overexpression of wild-type (WT) TRAF2 significantly protected cells from radiation-induced killing. Likewise, siRNA silencing of TRAF2 in radioresistant lung cancer H1299 cells caused growth suppression and radiosensitization, whereas overexpression of WT TRAF2 enhanced radioresistance in a RING ligase-dependent manner. Moreover, siRNA silencing of TRAF2 in UM-SCC-1 head and neck cancer cells also conferred radiosensitization. Further support for the role of TRAF2 in cancer comes from the observations that TRAF2 is overexpressed in both lung adenocarcinoma tissues and multiple lung cancer cell lines. Importantly, TRAF2 expression was very low in normal bronchial epithelial NL20 cells, and TRAF2 silencing had a minimal effect on NL20 growth and radiation sensitivity. Mechanistically, TRAF2 silencing blocks the activation of the nuclear factor-kappaB signaling pathway and down-regulates several G(2)-M cell cycle control proteins, resulting in enhanced G(2)-M arrest, growth suppression, and radiosensitization. Our studies suggest that TRAF2 is an attractive drug target for anticancer therapy and radiosensitization.


Inventory of surveillance systems assessing dietary, physical activity and sedentary behaviours in Europe: a DEDIPAC study.
There is a need for harmonized public health surveillance systems to monitor regional variations and temporal trends of health behaviours and health outcomes and to align policies, action plans and recommendations in terms of healthy diet and physical (in)activity within Europe. We provide an inventory of currently existing surveillance systems assessing diet, physical activity, and sedentary behaviours in Europe as a tool to assist in the identification of gaps and needs and to contribute to the roadmap for an integrated pan-European surveillance system. An inventory questionnaire was completed by representatives of eleven European countries. Eligible surveillance systems were required to meet specific inclusion criteria. First, pre-screening of available surveillance systems in each country was conducted. Second, an in-depth appraisal of the retained surveillance systems complying with the pre-defined requirements was performed. Fifty surveillance systems met the inclusion criteria: six multinational European surveys and forty-four national surveys. Dietary intake and physical activity are the domains predominantly assessed and adults are the most frequently studied age group. Many on-going activities were identified at the national level focussing on adults, but fewer surveillance systems involving vulnerable groups such as infants and pre-school children. Assessment of sedentary and dietary behaviours should be more frequently considered. There is a need for harmonization of surveillance methodologies, indicators and target populations for between-country and over time comparisons. This inventory will serve to feed future discussions within the DEDIPAC-JPI major framework on how to optimize design and identify priorities within surveillance.


Heterogeneous combinatorial expression of Hoxd genes in single cells during limb development.
Global analyses of gene expression during development reveal specific transcription patterns associated with the emergence of various cell types, tissues, and organs. These heterogeneous patterns are instrumental to ensure the proper formation of the different parts of our body, as shown by the phenotypic effects generated by functional genetic approaches. However, variations at the cellular level can be observed within each structure or organ. In the developing mammalian limbs, expression of Hox genes from the HoxD cluster is differentially controlled in space and time, in cells that will pattern the digits and the forearms. While the Hoxd genes broadly share a common regulatory landscape and large-scale analyses have suggested a homogenous Hox gene transcriptional program, it has not previously been clear whether Hoxd genes are expressed together at the same levels in the same cells. We report a high degree of heterogeneity in the expression of the Hoxd11 and Hoxd13 genes. We analyzed single-limb bud cell transcriptomes and show that Hox genes are expressed in specific combinations that appear to match particular cell types. In cells giving rise to digits, we find that the expression of the five relevant Hoxd genes (Hoxd9 to Hoxd13) is unbalanced, despite their control by known global enhancers. We also report that specific combinatorial expression follows a pseudo-time sequence, which is established based on the transcriptional diversity of limb progenitors. Our observations reveal the existence of distinct combinations of Hoxd genes at the single-cell level during limb development. In addition, we document that the increasing combinatorial expression of Hoxd genes in this developing structure is associated with specific transcriptional signatures and that these signatures illustrate a temporal progression in the differentiation of these cells.


An iron variant of the Noyori hydrogenation catalyst for the asymmetric transfer hydrogenation of ketones.
We report the design of a new iron catalyst for the asymmetric transfer hydrogenation of ketones. This type of iron catalyst combines the structural characteristics of the Noyori hydrogenation catalyst (an axially chiral 2,2'-bis(phosphino)-1,1'-binaphthyl fragment and the metal-ligand bifunctional motif) and an ene(amido) group that can activate the iron center. After activation by 8 equivalents of potassium tert-butoxide, (SA,RP,SS)-7a and (SA,RP,SS)-7b are active but nonenantioselective catalysts for the transfer hydrogenation of acetophenone and α,β-unsaturated aldehydes at room temperature in isopropanol. A maximum turnover number of 14480 was observed for (SA,RP,SS)-7a in the reduction of acetophenone. The right combination of the stereochemistry of the axially chiral 2,2'-bis(phosphino)-1,1'-binaphthyl group and the carbon-centered chiral amine-imine moiety in (SA,RP,RR)-7b' afforded an enantioselective catalyst for the preparation of chiral alcohols with moderate to good yields and a broad functional group tolerance.


Dropping Behavior in the Pea Aphid (Hemiptera: Aphididae): How Does Environmental Context Affect Antipredator Responses?
The pea aphid Acyrthosiphon pisum : Harris (Hemiptera: Aphididae) is a phloem-feeding insect whose antipredator defenses include kicking, walking away, and dropping from the plant. Aphid dropping, a risky and energetically costly antipredator behavior, can be increased by the release of aphid alarm pheromone; there is also evidence that insect density and plant health can affect the likelihood of aphids engaging in this behavior. We investigated whether interactions between alarm cues, insect density, and plant health can alter the dropping behavior of aphids in response to an artificial disturbance. The presence of the alarm pheromone E-β-farnesene resulted in a nearly 15-fold increase in aphid dropping behavior; the other two factors, however, did not affect dropping and none of the two- or three-way interactions were significant. This was surprising because aphids affected plant health: production of new plant biomass after 5 d of exposure to high aphid densities was 50% lower than in the control treatment. This research adds to our understanding of the factors affecting aphid antipredator behavior; the fact that neither aphid density nor feeding period impacted dropping may reflect the high energetic costs of this activity and an unwillingness to use it in any but the riskiest situations.


Differentiation of the 1-(methylenedioxyphenyl)-2-piperazinopropanes and 1-(methoxyphenyl)-2-piperazinopropanones by GC-IRD and GC-MS.
Two amphetamine-like piperazine-containing compounds, 1-(3,4-methylenedioxyphenyl)-2-piperazinopropane (3,4-MDPPP), its positional isomer 1-(2,3-methylenedioxyphenyl)-2-piperazinopropane (2,3-MDPPP) and three methcathinone-like piperazine-containing regioisomeric ring substituted 1-(methoxyphenyl)-2-piperazinopropanones (OMePPPOs) have identical elemental composition and no marked differences in their mass spectra. Perfluoroacylation of the secondary amine nitrogen of these isomeric piperazines gave mass spectra with differences in the relative abundance of some fragment ions but did not alter the fragmentation pathway to provide unique ions for discrimination among these isomers. Gas chromatography coupled to infrared detection (GC-IRD) provides direct confirmatory data for the identification of the carbonyl containing compounds and the differentiation of the 3,4-MDPPP from its direct (2,3-MDPPP) and indirect (OMePPPOs) regioisomers. The vapor phase infrared spectra provide for specific confirmation of each of the isomeric piperazines. The perfluoroacyl derivative forms of the five piperazines involved in this study were resolved on two stationary phases, the first is composed of 100% dimethyl polysiloxane (Rtx-1) and the second of 5% diphenyl and 95% dimethyl polysiloxane (Rtx-5).


Psychometric Properties of the Physical Educators' Self-Efficacy Toward Including Students With Disabilities-Autism Among Chinese Preservice Physical Education Teachers.
Teachers' self-efficacy is a critical predictor for successful inclusive physical education. However, little is known about preservice physical educators' self-efficacy toward teaching students with autism spectrum disorders in China. A sound instrument is necessary to measure their self-efficacy level. This validation study examined the psychometric properties of the Chinese version of the Physical Educators' Self-Efficacy Toward Including Students with Disabilities-Autism. A multisection survey form was administered to preservice physical educators in Mainland China (n = 205) and Hong Kong (n = 227). The results of confirmatory factor analysis confirmed the one-factor model of the scale in the total sample and each of the two samples. Invariance tests across the two samples supported configural and metric invariance but not scalar invariance. The scale scores showed good internal reliability and were correlated with theoretically relevant constructs (i.e., burnout and life satisfaction) in the total sample and subsamples. These findings generally support the utility of the scale for use among Chinese preservice physical educators.


Usefulness of 3 Tesla Ultrashort Echo Time Magnetic Resonance Angiography (UTE-MRA, SILENT-MRA) for Evaluation of the Mother Vessel after Cerebral Aneurysm Clipping: Case Series of 19 Patients.
It is important to assess the cerebral arteries near the clip after cerebral aneurysm clipping. Contrast-enhanced computed tomography angiography has side effects of contrast medium and radiation exposure. Time-of-flight magnetic resonance angiography (TOF-MRA) is a fast and non-invasive method, but clip-induced artifact limits the assessment around the clip. Recently, 3 tesla MRA with ultrashort echo time called SILENT MRA (GE Healthcare Life Sciences, UK) has been reported to have the potential to overcome these disadvantages. We herein present consecutive 19 cerebral aneurysm patients treated by clipping and evaluated using SILENT MRA. The 19 patients (15 women and 4 men) underwent TOF-MRA and SILENT MRA during the same scan session. Two neurosurgeons independently assessed the visibility of the mother vessel at the clipping site in TOF-MRA and SILENT MRA. We also investigated the factors related to visibility in SILENT MRA. All patients' mother vessels were not described in TOF-MRA, and that of 16 patients (84%) were described in SILENT MRA. Overall agreement was 100% in the two neurosurgeons, and the fixed marginal kappa = 1.00 (95% CI: 0.36-1.00). Univariate analysis revealed that larger aneurysm dome and long clip blade length contributed to the visibility of the mother vessel in SILENT MRA. (p = 0.023, 0.007, each). In conclusion, SILENT MRA can be applied for the assessment of the arteries and aneurysm neck remnants near the clip. Using clips with long blade and ligation with its tip would be related to the visibility of the mother vessels in SILENT MRA.


EQ-5D-5L population norms and health inequalities for Trinidad and Tobago.
The EQ-5D instrument is now used in many health systems as a health outcomes measure. Recently an EQ-5D valuation study was conducted for Trinidad and Tobago, but thus far there have been no population norms published for Trinidad and Tobago or for any Caribbean country. The objective of this study is to provide a set of population norms, and to investigate inequalities in health in Trinidad and Tobago. The EQ-5D-5L questionnaire was included in the 2012/2013 Adult Population Survey of the Global Entrepreneurship Monitor for Trinidad and Tobago. This survey covered a representative sample of 2,036 adults aged 18 and over. Demographic data and self-reported health using EQ-5D-5L were collected. The Trinidad and Tobago value set was used to obtain EQ-5D index values. The Kakwani index and logistic regression models were used to evaluate inequalities in health. Mean EQ-5D index values and EQ-VAS values were calculated by age group, ethnicity, gender, income, educational attainment, employment status and place of residence. The 10 most commonly observed EQ-5D-5L states accounted for 90% of the respondents. The mean VAS value for the sample was 83.6 and the mean EQ-5D-5L index value was 0.95. Pain/discomfort was found to be the EQ-5D dimension with the highest prevalence of reported problems with 22% of the population reporting pain at any level. Self-care was the dimension with the lowest prevalence of problems reported at any level (3%). Health declines with increasing age, and men reported fewer problems and higher levels of self-reported health than women. Age, gender and education level were found to be important drivers of health status as measured by the EQ-5D instrument. Being in a very low income group was also observed to affect EQ-VAS values among younger respondents. The population norms provided in this study can be used by clinicians, academics and policy makers in several ways. They can be used in comparing different demographic groups or patient groups, or as a basis for tracking the progress of patients through a treatment regimen. They can also provide a baseline for cost utility analysis of health interventions for Trinidad and Tobago.


Anterior dynamic ultrasound and Graf's examination in neonatal hip instability.
Discrepancy between neonatal hip morphology and stability has been reported in the literature. Comparative ultrasound studies on this issue are limited. To compare neonatal hip instability, as assessed by dynamic ultrasound and clinical examination, with acetabular morphology, as assessed by Graf's method. 536 newborn infants with clinical signs of hip instability, ambiguous findings at clinical hip examination, or positive risk factors for DDH were investigated with two ultrasound methods, the Graf method and anterior dynamic ultrasound, at an average age of 12 days. The hips were allocated to three groups according to the Graf result: A, normal (type Ia and b); B, borderline or immature (type IIa); and C, pathologic (type IIc and worse). Graf examination was compared with two diagnostic tests for instability, namely clinical examination by senior pediatric orthopedists and anterior dynamic ultrasound. According to Graf's method, 77% of the hips were normal, 20% borderline/immature, and 3% pathologic. On clinical hip examination, 82% were stable, 14% unstable, and 4% dislocatable. The dynamic ultrasound outcome was 88% stable hips, 10% unstable, and 2% dislocatable. Of the hips considered unstable or dislocatable on dynamic ultrasound, 21% had normal (type I) and 66% immature acetabular morphology according to the Graf method. Of the hips that were stable on dynamic ultrasound, only one (0.1%) was dysplastic according to the Graf method. Graf's examination showed the smallest number of normal hips, but also the fewest pathologic hips, with many indeterminate results that needed follow-up. Acetabular morphology correlated better to stability as assessed by dynamic ultrasound than to the clinical examination results, with fair to moderate agreement. Graf's examination resulted in a large number of indeterminate results that needed follow-up, but when used as the sole criterion for deciding treatment did not lead to a higher treatment rate than when the decision was based on clinical hip examination.


Breast tumor specific mutation in GATA3 affects physiological mechanisms regulating transcription factor turnover.
The transcription factor GATA3 is a favorable prognostic indicator in estrogen receptor-α (ERα)-positive breast tumors in which it participates with ERα and FOXA1 in a complex transcriptional regulatory program driving tumor growth. GATA3 mutations are frequent in breast cancer and have been classified as driver mutations. To elucidate the contribution(s) of GATA3 alterations to cancer, we studied two breast cancer cell lines, MCF7, which carries a heterozygous frameshift mutation in the second zinc finger of GATA3, and T47D, wild-type at this locus. Immunofluorescence staining and subcellular fractionation were employed to verify cellular localization of GATA3 in T47D and MCF7 cells. To test protein stability, cells were treated with translation inhibitor, cycloheximide or proteasome inhibitor, MG132, and GATA3 abundance was measured over time using immunoblot. GATA3 turn-over in response to hormone was determined by treating the cells with estradiol or ERα agonist, ICI 182,780. DNA binding ability of recombinant GATA3 was evaluated using electrophoretic mobility shift assay and heparin chromatography. Genomic location of GATA3 in MCF7 and T47D cells was assessed by chromatin immunoprecipitation coupled with next-generation sequencing (ChIP-seq). GATA3 localized in the nucleus in T47D and MCF7 cells, regardless of the mutation status. The truncated protein in MCF7 had impaired interaction with chromatin and was easily released from the nucleus. Recombinant mutant GATA3 was able to bind DNA to a lesser degree than the wild-type protein. Heterozygosity for the truncating mutation conferred protection from regulated turnover of GATA3, ERα and FOXA1 following estrogen stimulation in MCF7 cells. Thus, mutant GATA3 uncoupled protein-level regulation of master regulatory transcription factors from hormone action. Consistent with increased protein stability, ChIP-seq profiling identified greater genome-wide accumulation of GATA3 in MCF7 cells bearing the mutation, albeit with a similar distribution across the genome, comparing to T47D cells. We propose that this specific, cancer-derived mutation in GATA3 deregulates physiologic protein turnover, stabilizes GATA3 binding across the genome and modulates the response of breast cancer cells to estrogen signaling.


Taking off the gloves: toward a less dogmatic approach to the use of contact isolation.
Strongly held beliefs about the need for contact isolation to prevent the spread of infections in the hospital have contributed to increased costs and decreased flexibility and, more recently, have driven aggressive diagnostic testing for colonization in asymptomatic patients. Examination of the evidence cited in support of the benefits of isolation and growing evidence of its unintended harms offer an opportunity to think differently about how contact isolation might best be applied. This review considers what we do and do not know about the potential benefits and harms of isolation as a public health measure and proposes a framework for considering under what circumstances it might optimally be used.


Learning-based structurally-guided construction of resting-state functional correlation tensors.
Functional magnetic resonance imaging (fMRI) measures changes in blood-oxygenation-level-dependent (BOLD) signals to detect brain activities. It has been recently reported that the spatial correlation patterns of resting-state BOLD signals in the white matter (WM) also give WM information often measured by diffusion tensor imaging (DTI). These correlation patterns can be captured using functional correlation tensor (FCT), which is analogous to the diffusion tensor (DT) obtained from DTI. In this paper, we propose a noise-robust FCT method aiming at further improving its quality, and making it eligible for further neuroscience study. The novel FCT estimation method consists of three major steps: First, we estimate the initial FCT using a patch-based approach for BOLD signal correlation to improve the noise robustness. Second, by utilizing the relationship between functional and diffusion data, we employ a regression forest model to learn the mapping between the initial FCTs and the corresponding DTs using the training data. The learned forest can then be applied to predict the DTI-like tensors given the initial FCTs from the testing fMRI data. Third, we re-estimate the enhanced FCT by utilizing the DTI-like tensors as a feedback guidance to further improve FCT computation. We have demonstrated the utility of our enhanced FCTs in Alzheimer's disease (AD) diagnosis by identifying mild cognitive impairment (MCI) patients from normal subjects.


Management of peripherally inserted central catheters (PICC) in pediatric heart failure patients receiving continuous inotropic support.
The study aim was to evaluate present practice of maintaining PICC line patency in pediatric heart failure patients receiving continuous inotropes by comparing one cohort receiving low dose continuous heparin with one receiving no heparin. A case control retrospective chart review compared the two cohorts on duration of patency (measured in days) and need for thrombolytic agents. Median duration of patency for the heparin group was 24 days versus 16 days for the no heparin group (p=0.07). Use of thrombolytic agents was 28% in the heparin group compared to 50% in the no heparin group (p=0.08). Although not statistically significant, findings were clinically significant and supportive of current practice.


Neocortical dendritic complexity is controlled during development by NOMA-GAP-dependent inhibition of Cdc42 and activation of cofilin.
Neocortical neurons have highly branched dendritic trees that are essential for their function. Indeed, defects in dendritic arborization are associated with human neurodevelopmental disorders. The molecular mechanisms regulating dendritic arbor complexity, however, are still poorly understood. Here, we uncover the molecular basis for the regulation of dendritic branching during cortical development. We show that during development, dendritic branching requires post-mitotic suppression of the RhoGTPase Cdc42. By generating genetically modified mice, we demonstrate that this is catalyzed in vivo by the novel Cdc42-GAP NOMA-GAP. Loss of NOMA-GAP leads to decreased neocortical volume, associated specifically with profound oversimplification of cortical dendritic arborization and hyperactivation of Cdc42. Remarkably, dendritic complexity and cortical thickness can be partially restored by genetic reduction of post-mitotic Cdc42 levels. Furthermore, we identify the actin regulator cofilin as a key regulator of dendritic complexity in vivo. Cofilin activation during late cortical development depends on NOMA-GAP expression and subsequent inhibition of Cdc42. Strikingly, in utero expression of active cofilin is sufficient to restore postnatal dendritic complexity in NOMA-GAP-deficient animals. Our findings define a novel cell-intrinsic mechanism to regulate dendritic branching and thus neuronal complexity in the cerebral cortex.


Flow cytometric analyses of antibody binding to Chinese hamster ovary cells expressing human thyrotropin receptor.
To develop a method that can be used to directly detect binding of antibodies to TSH receptor (TSHr), we employed Chinese hamster ovary (CHO) cells permanently transfected with a human TSHr complementary DNA (CHOR). These cells showed increased cAMP production when treated with either human TSH or thyroid-stimulating antibodies and decreased TSH-mediated cAMP production when treated with stimulation-blocking antibodies. We employed flow cytometry and rabbit antibodies against the extracellular domain of the TSHr (ETSHr) to test whether these cells can be used to directly detect and quantitate the binding of anti-TSHr antibodies. Rabbit anti-ETSHr bound specifically to CHOR cells, and the binding could be blocked with purified ETSHr. To test the feasibility of using these cells for epitope mapping, we tested the binding of rabbit antibodies raised against several synthetic TSHr peptides. Rabbit antipeptide 92 (amino acids 12-30) and 91 (amino acids 32-46) showed little or no binding to the CHOR cells. In contrast, antibodies raised against peptides 93 (amino acids 316-330), 95 (aa 325-345), 3A (aa 357-372), 367 (aa 367-386), and 1B (aa 362-376) showed significant binding to the CHOR cells. The specificity of binding of antipeptide antibodies was demonstrated by a complete inhibition of binding by corresponding peptides. When TSH-binding inhibitory Ig-positive sera from 15 patients with hyperthyroidism were tested, 8 of them showed specific binding to the CHOR cells compared to their relative binding to normal CHO cells; sera from all normal individuals tested did not exhibit specific binding to CHOR cells. These studies showed the usefulness of CHOR cells and flow cytometry in epitope mapping using sera with known specificities and the potential usefulness of the technique to detect anti-TSHr antibodies in patient sera.


Exercise-induced expression of heme oxygenase-1 in human lymphocytes.
The aim of the present investigation was to determine whether an acute bout of exercise increases heme oxygenase-1 (HO-1) mRNA accumulation in human lymphocytes. Eight male subjects performed separate exercise and rest trials in a randomised order at least 10 days apart. In the exercise trial subjects ran for 75-min at a speed corresponding to 70% maximal oxygen uptake, and in the resting trial subjects sat calmly in the laboratory for an equivalent period of time. Lymphocytes were harvested from blood samples taken before and after each trial. Total RNA was isolated and used to determine the fold-change in HO-1 mRNA accumulation relative to baseline values using real time reverse transcription-polymerase chain reaction. HO-1 protein was determined by Western blotting. Six of the eight subjects showed an increase in HO-1 mRNA greater than two-fold after exercise. The median peak fold-change was 2.7 fold with one subject showing a particularly pronounced response (20-fold) 24 h post-exercise. In the rest trial the level of HO-1 mRNA did not change over the period of investigation. There was also an increase in HO-1 protein 2 h after exercise. These results complement an earlier study showing that acute exercise of a different type (half marathon) leads to an increase in HO-1 expression in lymphocytes.