Cytochrome P450IIIA enzymes in rat liver microsomes: involvement in C3-hydroxylation of diazepam and nordazepam but not N-dealkylation of diazepam and temazepam.
Microsomes prepared from livers of male and female rats of nine inbred and outbred strains and of male Sprague-Dawley rats pretreated with monooxygenase-inducing agents were used to study N-dealkylation of diazepam and temazepam and C3-hydroxylation of diazepam and nordazepam. Both C3-hydroxylation reactions were more rapid in male than in female liver preparations, but this gender-dependent pattern was not seen with the N-dealkylation reactions. These results indicate the lack of identity of the monooxygenases responsible for the two kinds of reaction and suggest that male-specific enzyme(s) are responsible for the C3-hydroxylations. Induction studies were undertaken to further define these enzymes. To do this, liver microsomes prepared from male Sprague-Dawley rats pretreated with a variety of agents known to have different monooxygenase induction effects were used. With triacetyloleandomycin, dexamethasone, and phenobarbital pretreatment, the specific activities of the C3-hydroxylation reactions were selectively elevated over corresponding control values. These particular xenobiotics are known to enhance the abundance of cytochrome P450IIIA family enzymes, and our results strongly suggest the involvement of these enzymes in the benzodiazepine B ring monooxygenations. Formation of temazepam was also shown to be inhibited by triacetyloleandomycin. This effect was demonstrated to be equal in both saline-treated and dexamethasone-treated male Sprague-Dawley rat liver microsomes, with the antibiotic present either with diazepam throughout the entire incubation period or initially with NADPH in a preincubation mix for 15 min, following which C3-hydroxylation was initiated by the addition of diazepam. These results confirm the uniformity of the involvement of cytochrome P450IIIA family enzymes in diazepam C3-hydroxylation in untreated and inducer-treated rat liver microsomes. Recent studies with human and rabbit liver microsomal preparations have shown that orthologues of these enzymes also catalyze an equivalent hydroxylation in the B ring of midazolam. These findings, considered with the present results showing that the adjacent methyl N-substituent (absent in nordazepam but present in diazepam) did not affect the selectivity of these enzymes for the C3-hydroxylation reaction, suggest that neither steric nor electronic factors markedly influence catalysis of this monooxygenation by these enzymes.


Crystal structure of the low-pH form of the vesicular stomatitis virus glycoprotein G.
The vesicular stomatitis virus has an atypical membrane fusion glycoprotein (G) exhibiting a pH-dependent equilibrium between two forms at the virus surface. Membrane fusion is triggered during the transition from the high- to low-pH form. The structure of G in its low-pH form shows the classic hairpin conformation observed in all other fusion proteins in their postfusion conformation, in spite of a novel fold combining features of fusion proteins from classes I and II. The structure provides a framework for understanding the reversibility of the G conformational change. Unexpectedly, G is homologous to gB of herpesviruses, which raises important questions on viral evolution.


Detection of KRAS G12/G13 Mutations in Cell Free-DNA by Droplet Digital PCR, Offers Prognostic Information for Patients with Advanced Non-Small Cell Lung Cancer.
KRAS mutations are found in approximately one third of non-small cell lung cancer (NSCLC) patients. In this study, we aim to investigate whether KRAS G12/G13 mutant allele fraction (MAF) in cell-free DNA (cfDNA) can provide meaningful prognostic information in NSCLC. Multiplex droplet-digital PCR was used to quantitatively assess KRAS G12/G13 MAF in cfDNA from 114 pre-treated advanced disease NSCLC patients. In 14 patients, changes in KRAS G12/G13 MAF were longitudinally monitored during treatment. Plasma KRAS G12/G13 status was associated with poor patients' outcome in terms of progression-free survival (PFS) (p < 0.001) and overall survival (OS) (p < 0.001). In multivariate analysis, the detection of plasma KRAS mutations was an independent predictor of adverse PFS (HR = 3.12; p < 0.001) and OS (HR = 2.53; p = 0.002). KRAS G12/G13 MAF at first treatment evaluation (T1) was higher (p = 0.013) among patients experiencing progressive disease compared to those with disease control, and increased KRAS MAF at T1 was associated (p = 0.005) with shorter PFS. On the contrary, no association was observed between tissue KRAS mutation status and patients' prognosis. Our results show that ddPCR-based detection of KRAS G12/G13 mutations in plasma could serve as an independent biomarker of unfavorable prognosis in NSCLC patients. Changes in KRAS MAF can provide valuable information for monitoring patient outcome during treatment.


Characterization of the HLA-A2.2 subtype: T cell evidence for further heterogeneity.
Five blood donors were identified whose HLA-A2 is different from the common HLA-A2. Their A2 molecule (A2.2) had a more basic isoelectric point than normal A2 (A2.1). Cytotoxic T lymphocytes (CTL) restricted by HLA-A2.1, specific for influenza A and Epstein-Barr viruses, failed to lyse virus-infected target cells with HLA-A2.2. Identical patterns were obtained with both viruses. CTL from four of the A2.2-positive donors recognized target cells prepared from others in the group that shared only the HLA-A2.2 antigen. The A2.2 antigen from one donor seemed to be different in that target cells were not recognized by CTL from donors with the normal A2.1 nor with basic A2.2. There seems, therefore, to be heterogeneity within the HLA-A2.2 subtype.


Rat DNA sequence associated with a complex form of DNA polymerase alpha in nonregenerating liver interacts with a ubiquitous transcription/replication factor Oct-1.
A ubiquitous mammalian transcription factor, Oct-1 (also known as OTF-1, NF-A1, OBP100, or NFIII), stimulates the initiation of replication of adenovirus DNA, and may also be involved in the activation of some chromosomal replication origins. If this is true, binding sites for Oct-1 should be present within regions responsible for the initiation of DNA replication. In this study such a binding site has been identified within a 340bp fragment that was originally isolated from a minor fraction of DNA associated with a complexed form of DNA polymerase alpha from nonregenerating rat liver, and which shows autonomous replication sequence activity in a transient transfection assay. Northern blot analysis was used to show that Oct-1 mRNA is induced in regenerating rat liver 6-14 h after hepatectomy.


Genetic aspects of behavioral neurotoxicology.
Considerable progress has been made over the past couple of decades concerning the molecular bases of neurobehavioral function and dysfunction. The field of neurobehavioral genetics is becoming mature. Genetic factors contributing to neurologic diseases such as Alzheimer's disease have been found and evidence for genetic factors contributing to other diseases such as schizophrenia and autism are likely. This genetic approach can also benefit the field of behavioral neurotoxicology. It is clear that there is substantial heterogeneity of response with behavioral impairments resulting from neurotoxicants. Many factors contribute to differential sensitivity, but it is likely that genetic variability plays a prominent role. Important discoveries concerning genetics and behavioral neurotoxicity are being made on a broad front from work with invertebrate and piscine mutant models to classic mouse knockout models and human epidemiologic studies of polymorphisms. Discovering genetic factors of susceptibility to neurobehavioral toxicity not only helps identify those at special risk, it also advances our understanding of the mechanisms by which toxicants impair neurobehavioral function in the larger population. This symposium organized by Edward Levin and Annette Kirshner, brought together researchers from the laboratories of Michael Aschner, Douglas Ruden, Ulrike Heberlein, Edward Levin and Kathleen Welsh-Bohmer conducting studies with Caenorhabditis elegans, Drosophila, fish, rodents and humans studies to determine the role of genetic factors in susceptibility to behavioral impairment from neurotoxic exposure.


Optimal beams for propagation through random media.
The problem of maximizing the intensity that is transferred from a transmitter aperture to a receiver aperture is considered in which the propagation medium is random. Two optimization criteria are considered: maximal expected intensity transfer and minimal scintillation index. The beam that maximizes the expected intensity is shown to be fully coherent. Its coherent mode is determined as the principal eigenfunction for a kernel that is determined through the second-order moments of the propagation Green's function. The beam that minimizes the scintillation index is shown to be partially coherent in general, with its coherent modes determined by minimizing a quadratic form that has nonlinear dependence on the coherent-mode fields, and on the second- and fourth-order moments of the propagation Green's function.


Perceived needs in women with gestational diabetes: A qualitative study.
Diabetes is the most common medical complication of pregnancy. It can be associated with many complications for mother and fetus. Gestational diabetes is also one of the main health issues in Iran. Therefore, the present study is aimed at a deeper understanding of women's experiences of gestational diabetes and their perceived needs to inform future lifestyle interventions. This qualitative content analysis study was carried out in 2015. Participants were pregnant women diagnosed with gestational diabetes in the 24th to 36th week of pregnancy, who were referred to the clinics affiliated with Shahid Beheshti Medical Science University in Tehran, Iran. In-depth interviews were conducted with participants, using semi-structured questions. Interviews were audio taped and transcribed verbatim. Conventional content analysis was carried out for data analysis. Interviews continued until data saturation was obtained. Data were coded in MAXQDA software (version 11). Content analysis highlighted two themes; educational needs and need to support. The former was featured with five main categories: information sources, education process, unknown and known, weaknesses of public information system, and eagerness to learn. The latter was featured with two main categories: family support and social support. Clarifying the needs of the mothers with gestational diabetes, leads to better and proper education planning and a program toward the improvement of health, self-care, and prevention of diabetes.


Emergent Multifunctional Magnetic Proximity in van der Waals Layered Heterostructures.
Proximity effect, which is the coupling between distinct order parameters across interfaces of heterostructures, has attracted immense interest owing to the customizable multifunctionalities of diverse 3D materials. This facilitates various physical phenomena, such as spin order, charge transfer, spin torque, spin density wave, spin current, skyrmions, and Majorana fermions. These exotic physics play important roles for future spintronic applications. Nevertheless, several fundamental challenges remain for effective applications: unavoidable disorder and lattice mismatch limits in the growth process, short characteristic length of proximity, magnetic fluctuation in ultrathin films, and relatively weak spin-orbit coupling (SOC). Meanwhile, the extensive library of atomically thin, 2D van der Waals (vdW) layered materials, with unique characteristics such as strong SOC, magnetic anisotropy, and ultraclean surfaces, offers many opportunities to tailor versatile and more effective functionalities through proximity effects. Here, this paper focuses on magnetic proximity, i.e., proximitized magnetism and reviews the engineering of magnetism-related functionalities in 2D vdW layered heterostructures for next-generation electronic and spintronic devices. The essential factors of magnetism and interfacial engineering induced by magnetic layers are studied. The current limitations and future challenges associated with magnetic proximity-related physics phenomena in 2D heterostructures are further discussed.


Evaluation of resistance in 8 different heat-and-moisture exchangers: effects of saturation and flow rate/profile.
When endotracheal intubation is required during ventilatory support, the physiologic mechanisms of heating and humidifying the inspired air related to the upper airways are bypassed. The task of conditioning the air can be partially accomplished by heat-and-moisture exchangers (HMEs). To evaluate and compare with respect to imposed resistance, different types/models of HME: (1) dry versus saturated, (2) changing inspiratory flow rates. Eight different HMEs were studied using a lung model system. The study was conducted initially by simulating spontaneous breathing, followed by connecting the system directly to a mechanical ventilator to provide pressure-support ventilation. None of the encountered values of resistance (0.5\N3.6 cm H(2)O/L/s) exceeded the limits stipulated by the previously described international standard for HMEs (International Standards Organization Draft International Standard 9360-2) (not to exceed 5.0 cm H(2)O with a flow of 1.0 L/s, even when saturated). The hygroscopic HME had less resistance than other types, independent of the precondition status (dry or saturated) or the respiratory mode. The hygroscopic HME also had a lesser increase in resistance when saturated. The resistance of the HME was little affected by increases in flow, but saturation did increase resistance in the hydrophobic and hygroscopic/hydrophobic HME to levels that could be important at some clinical conditions. Resistance was little affected by saturation in hygroscopic models, when compared to the hydrophobic or hygroscopic/hydrophobic HME. Changes in inspiratory flow did not cause relevant alterations in resistance.


Uptake by COPI-coated vesicles of both anterograde and retrograde cargo is inhibited by GTPgammaS in vitro.
On the basis of the cell surface protein CD8 we have constructed reporter molecules for both anterograde and retrograde transport from the Golgi complex. The cytoplasmic tail of CD8 was exchanged by a construct comprising a hemagglutinin (HA) epitope, the C-terminal sequence of the viral protein E19 (containing a KKXX retrieval signal) followed by a myc epitope (CD8-LT). Due to this masking of the KKXX retrieval signal CD8-LT is transported to the cell surface. Since the KKXX motif is joined to the myc epitope via a thrombin cleavage site, CD8-LT in isolated Golgi membranes can be proteolytically converted into an unmasked reporter molecule for retrograde transport (CD8-ST) in vitro. A CHO cell line stably expressing CD8-LT was generated and used for the isolation of Golgi membranes. These membranes were shown to contain CD8-LT en route to the cell surface. By addition of thrombin, CD8-LT could be efficiently converted into CD8-ST, and this allows us to study the sorting into coat protein COPI-coated vesicles of these different kinds of cargo on a comparative basis. COPI-coated vesicles were generated in vitro from Golgi membranes containing either CD8-LT or CD8-ST. When the incubation was performed in the presence of GTP, both CD8-LT and CD8-ST were packaged into COPI-coated vesicles. However, COPI-coated vesicles generated in the presence of the slowly hydrolyzable analogue of GTP, GTP(&ggr ;)S contained strikingly lower amounts of CD8-LT and CD8-ST. While COPI-coated vesicles accumulated about 12-fold in the presence of GTPgammaS these vesicles together contained only one fifth of cargo compared to the few vesicles generated in the absence of GTPgammaS. These data indicate that cargo packaging into COPI-coated vesicles requires hydrolysis of GTP.


Optimizing chronic HIV care in the Dutch Caribbean: a questionnaire based study on experience and perception of healthcare workers.
Human immunodeficiency virus (HIV) infection has become a chronic disorder for which adaptation of current healthcare practices is needed. In Curaçao, a new organization of chronic HIV care is being set up based on task shifting in which healthcare workers (HCWs) will deliver HIV care more prominently within the primary healthcare system. In preparation for implementation of the proposed task-shifting model, we investigated the perception of HCWs regarding existing HIV care in Curaçao and the need for training in HIV/AIDS among HCWs. An in-depth questionnaire based study was used. Nineteen HCWs of seven different cadres were interviewed. The questionnaire constituted four sections: quality of existing HIV care, respondents own knowledge and willingness to be trained, need for training in HIV/AIDS and preferred educational approaches. Quality of existing HIV services in Curaçao is considered acceptable but needs improvement mainly to facilitate integration of chronic HIV care. All respondents indicated that training in HIV/AIDS is needed among HCWs in Curaçao, especially for nurses and general practitioners. All were willing to participate in training with varying amounts of time to be spent. Training should be tailored to the level of expertise of HCWs and to the role a HCW is expected to have in the new healthcare delivery framework. There is need for training to ensure the effective integration of chronic HIV care into the existing healthcare delivery system in Curaçao.There is a willingness and need in all cadres for training with e-learning as a preferred educational tool.


An investigation of antifungal stewardship programmes in England.
We sought to explore the current status of antifungal stewardship (AFS) initiatives across National Health Service (NHS) Trusts within England, the challenges and barriers, as well as ways to improve current AFS programmes. An electronic survey was sent to all 155 acute NHS Trusts in England. A total of 47 Trusts, corresponding to 30 % of English acute Trusts, responded to the the survey; 46 Trusts (98 %) had an antimicrobial stewardship (AMS) programme but only 5 (11 %) had a dedicated AFS programme. Overall, 20 (43 %) Trusts said they included AFS as part of their AMS programmes. From those conducting AFS programmes, 7 (28 %) have an AFS/management team, 16 (64 %) monitor and report on antifungal usage, 5 (20 %) have dedicated AFS ward rounds and 12 (48 %) are directly involved in the management of invasive fungal infections.Results/Key findings. Altogether, 13 acute Trusts (52 %) started their AFS programme to manage costs, whilst 12 (48 %) commenced the programme due to clinical need; 27 (73 %) declared that they would increase their AFS initiatives if they could. Of those without an AFS programme, 14 (67 %) responded that this was due to lack of resources/staff time. Overall, 12 Trusts (57 %) responded that the availability of rapid diagnostics and clinical support would enable them to conduct AFS activities. Although a minority of Trusts conduct dedicated AFS programmes, nearly half include AFS as part of routine AMS activities. Cost issues are the main driver for AFS, followed by clinical need. The availability of rapid diagnostics and clinical support could help increase AFS initiatives.


Adrenal venous sampling for primary aldosteronism: laboratory medicine best practice.
Primary aldosteronism (PA) is the most common form of secondary hypertension and is critical to identify because when caused by an aldosterone-producing adenoma (APA) or another unilateral form, it is potentially curable, and even when caused by bilateral disease, antihypertensives more specific to PA treatment can be employed (ie, aldosterone antagonists). Identification of unilateral forms is not generally accomplished with imaging because APAs may be small and elude detection, and coincidental identification of a non-functioning incidentaloma contralateral to an APA may lead to removal of an incorrect gland. For this reason, the method of choice for identifying unilateral forms of PA is selective adrenal venous sampling (AVS) followed by aldosterone and cortisol analysis on collected samples. This procedure is technically difficult from a radiological standpoint and, from the laboratory perspective, is fraught with opportunities for preanalytical, analytical and postanalytical error. We review the process of AVS collection, analysis and reporting. Suggestions are made for patient preparation, specimen labelling practices and nomenclature, analytical dilution protocols, which numerical results to report, and the necessary subsequent calculations. We also identify and explain frequent sources of confusion in the aldosterone and cortisol results and provide an example of tabular reporting to facilitate interpretation and communication between laboratorian, radiologist and clinician.


A missense mutation in the alphaB-crystallin chaperone gene causes a desmin-related myopathy.
Desmin-related myopathies (DRM) are inherited neuromuscular disorders characterized by adult onset and delayed accumulation of aggregates of desmin, a protein belonging to the type III intermediate filament family, in the sarcoplasma of skeletal and cardiac muscles. In this paper, we have mapped the locus for DRM in a large French pedigree to a 26-cM interval in chromosome 11q21-23. This region contains the alphaB-crystallin gene (CRYAB), a candidate gene encoding a 20-kD protein that is abundant in lens and is also present in a number of non-ocular tissues, including cardiac and skeletal muscle. AlphaB-crystallin is a member of the small heat shock protein (shsp) family and possesses molecular chaperone activity. We identified an R120G missense mutation in CRYAB that co-segregates with the disease phenotype in this family. Muscle cell lines transfected with the mutant CRYAB cDNA showed intracellular aggregates that contain both desmin and alphaB-crystallin as observed in muscle fibers from DRM patients. These results are the first to identify a defect in a molecular chaperone as a cause for an inherited human muscle disorder.


A simulation study on the statistical monitoring of condemnation rates from slaughterhouses for syndromic surveillance: an evaluation based on Swiss data.
Syndromic surveillance (SyS) systems currently exploit various sources of health-related data, most of which are collected for purposes other than surveillance (e.g. economic). Several European SyS systems use data collected during meat inspection for syndromic surveillance of animal health, as some diseases may be more easily detected post-mortem than at their point of origin or during the ante-mortem inspection upon arrival at the slaughterhouse. In this paper we use simulation to evaluate the performance of a quasi-Poisson regression (also known as an improved Farrington) algorithm for the detection of disease outbreaks during post-mortem inspection of slaughtered animals. When parameterizing the algorithm based on the retrospective analyses of 6 years of historic data, the probability of detection was satisfactory for large (range 83-445 cases) outbreaks but poor for small (range 20-177 cases) outbreaks. Varying the amount of historical data used to fit the algorithm can help increasing the probability of detection for small outbreaks. However, while the use of a 0·975 quantile generated a low false-positive rate, in most cases, more than 50% of outbreak cases had already occurred at the time of detection. High variance observed in the whole carcass condemnations time-series, and lack of flexibility in terms of the temporal distribution of simulated outbreaks resulting from low reporting frequency (monthly), constitute major challenges for early detection of outbreaks in the livestock population based on meat inspection data. Reporting frequency should be increased in the future to improve timeliness of the SyS system while increased sensitivity may be achieved by integrating meat inspection data into a multivariate system simultaneously evaluating multiple sources of data on livestock health.


Walnut ( Juglans regia) Peptides Reverse Sleep Deprivation-Induced Memory Impairment in Rat via Alleviating Oxidative Stress.
The aim of this study was to determine the neuroprotective effects of walnut protein hydrolysates (WPH) against memory deficits induced by sleep deprivation (SD) in rat and further to identify and characterize the potent neuroprotective peptides against glutamate-induced apoptosis in PC12 cells. Results showed that a remarkable amelioration effect on behavioral performance in Morris water maze test was observed for WPH and its low molecular weight fraction WPHL, especially for WPHL. Additionally, a reduction of antioxidant defense (catalase, glutathione peroxidase (GSH-px), and superoxide dismutase (SOD)) and an increase of malondialdehyde content induced by SD were normalized in brain of rat after oral administration of WPH and WPHL. Then three neuroprotective peptides including GGW, VYY, and LLPF were identified from WPHL, which could protect PC12 cells against glutamate-induced apoptosis with relative cell viability of 78.29 ± 3.09%, 80.65 ± 1.74%, and 83.97 ± 3.06%, respectively, versus glutamate group 48.61 ± 3.99%. The possible mechanism underlying their protective effects of GGW and VYY could be related to their strong radical scavenging activity as well as their ability to reduce reactive oxygen species production and the depletion of SOD and GSH-px in PC12 cells. Notably, the marked neuroprotective effects of LLPF, which did not show obvious free-radical scavenging activity in vitro, could be attributed to its strong effects on inhibiting Ca2+ influx and mitochondrial membrane potential collapse. Additionally, all these peptides could regulate the expression of apoptosis-related proteins (Bax and Bcl-2). Therefore, walnut peptides might be regarded as the potential nutraceuticals against neurodegenerative disorders associated with memory deficits.


Sequence and functional characterization of hypoxia-inducible factors, HIF1α, HIF2αa, and HIF3α, from the estuarine fish, Fundulus heteroclitus.
The hypoxia-inducible factor (HIF) family of transcription factors plays central roles in the development, physiology, pathology, and environmental adaptation of animals. Because many aquatic habitats are characterized by episodes of low dissolved oxygen, fish represent ideal models to study the roles of HIF in the response to aquatic hypoxia. The estuarine fish Fundulus heteroclitus is found in habitats prone to hypoxia. It responds to low oxygen via behavioral, physiological, and molecular changes, and one member of the HIF family, HIF2α, has been previously described. Herein, cDNA sequencing, phylogenetic analyses, and genomic approaches were used to determine other members of the HIFα family from F. heteroclitus and their relationships to HIFα subunits from other vertebrates. In vitro and cellular approaches demonstrated that full-length forms of HIF1α, HIF2α, and HIF3α independently formed complexes with the β-subunit, aryl hydrocarbon receptor nuclear translocator, to bind to hypoxia response elements and activate reporter gene expression. Quantitative PCR showed that HIFα mRNA abundance varied among organs of normoxic fish in an isoform-specific fashion. Analysis of the F. heteroclitus genome revealed a locus encoding a second HIF2α-HIF2αb-a predicted protein lacking oxygen sensing and transactivation domains. Finally, sequence analyses demonstrated polymorphism in the coding sequence of each F. heteroclitus HIFα subunit, suggesting that genetic variation in these transcription factors may play a role in the variation in hypoxia responses among individuals or populations.


Prenatal exposure to very severe maternal obesity is associated with adverse neuropsychiatric outcomes in children.
Prenatal maternal obesity has been linked to adverse childhood neuropsychiatric outcomes, including increased symptoms of attention deficit hyperactivity disorder (ADHD), internalizing and externalizing problems, affective disorders and neurodevelopmental problems but few studies have studied neuropsychiatric outcomes among offspring born to very severely obese women or assessed potential familial confounding by maternal psychological distress. We evaluated neuropsychiatric symptoms in 112 children aged 3-5 years whose mothers had participated in a longitudinal study of obesity in pregnancy (50 very severe obesity, BMI ⩾40 kg/m2, obese class III and 62 lean, BMI 18.5-25 kg/m2). The mothers completed the Conners' Hyperactivity Scale, Early Symptomatic Syndrome Eliciting Neurodevelopmental Clinical Examination Questionnaire (ESSENCE-Q), Child's Sleep Habits Questionnaire (CSHQ), Strengths and Difficulties Questionnaire (SDQ), and Child Behavior Checklist (CBCL) to assess child neuropsychiatric symptoms. Covariates included child's sex, age, birthweight, gestational age, socioeconomic deprivation levels, maternal age, parity, smoking status during pregnancy, gestational diabetes and maternal concurrent symptoms of anxiety and depression assessed using State Anxiety of Spielberger State-Trait Anxiety Index (STAI) and General Health Questionnaire (GHQ), respectively. Children exposed to prenatal maternal very severe obesity had significantly higher scores in the Conners' Hyperactivity Scale; ESSENCE-Q; total sleep problems in CSHQ; hyperactivity, conduct problems and total difficulties scales of the SDQ; higher externalizing and total problems, anxious/depressed, aggressive behaviour and other problem syndrome scores and higher DSM-oriented affective, anxiety and ADHD problems in CBCL. Prenatal maternal very severe obesity remained a significant predictor of child neuropsychiatric problems across multiple scales independent of demographic factors, prenatal factors and maternal concurrent symptoms of anxiety and depression. Prenatal maternal very severe obesity is a strong predictor of increased neuropsychiatric problems in early childhood.


The influence of inhibitors of apoptosis proteins (IAPs) on chronic rhinosinusitis with nasal polyps.
Inhibitors of apoptosis proteins (IAPs) modulate the inflammatory process, and may facilitate the formation of chronic rhinosinusitis with nasal polyps (CRSwNP). This study aimed to observe if IAPs were differently expressed between patients with CRSwNP and controls, and to correlate the expression of IAPs with some inflammatory markers, as with the response to nasal corticosteroids in patients with CRSwNP. We obtained nasal biopsies from patients with CRSwNP (n=27) and controls (n=16). qRT-PCR measured the expression of IAPs and caspases, while Luminex assay measured the concentration of cytokines. Unpaired parametric tests and Principal Component Analysis (PCA) were used for statistical analysis. We observed lower expression of IAP genes (XIAP, BIRC2/IAP1, and BIRC3/IAP2) in CRSwNP patients compared to controls, and we identified that patients with bad response to corticosteroids presented lower levels of BIRC2/IAP1, XIAP, BCL2, CASP9, and IL-17, and higher levels of CASP7 and TGF-β. IAPs expression was downregulated in CRSwNP, and was associated with poorer response to nasal corticosteroids. The present findings suggest the importance of IAPs as a link between environment and the host inflammatory responses, and this pathway could be explored as a potential new target therapy for patients with CRSwNP.


Clear benefit of mycophenolate mofetil-based triple therapy in reducing the incidence of acute rejection after living donor renal transplantations.
According to a pooled analysis of three randomized clinical studies concerning the prevention of acute rejection in cadaveric renal transplantation, mycophenolate mofetil (MMF) proved superior to azathioprine or placebo in conjunction with cyclosporine (CsA) and steroids. MMF-treated patients showed reduced incidence and severity of acute rejection, similar graft survival, and better graft function over 12 months. However, the multicenter trials did not include the Asian recipients of living donor kidneys. To assess the efficacy of MMF as the third component of a triple therapy in addition to CsA-Neoral and steroids in living donor renal transplantation recipients in Asians, a total of 100 recipients were randomized to receive CsA-Neoral and steroids (control group, n=50), or MMF-based triple therapy (1.0 g of MMF twice daily from postoperative day 2, MMF group, n=50). The dosing plan for Neoral and steroids was essentially same between groups. During 12 months of follow-up, we compared the incidence of acute rejection, adverse events such as infections, and 12-month actual graft and patient survival. The graft and patient survival at 1 year was excellent in both groups: 96/98% in the control group and 98/100% in the MMF group, respectively. MMF significantly reduced the proportion of patients with at least one episode of acute rejection (34% in the control group vs. 14% in the MMF group), cumulative incidence of acute rejection episodes (46% vs. 16%), and requirement of antilymphocyte antibody (21.7% vs. 12.5%). In the MMF group, viral infection such as herpes zoster or chicken pox was more prevalent than in the control group. Like cadaveric renal transplantation, this open clinical trial showed MMF to be effective in reducing the incidence and severity of acute rejection if used in conjunction with Neoral and steroids after living donor renal transplantation in Asian ethnicity.


Caesalpinia sappan: A promising natural source of antimicrobial agent for inhibition of cariogenic bacteria.
From the previous findings, the ethanolic fractionated extract of Caesalpinia sappan (F-EtOH) has high activity against Streptococcus mutans, the most severe cariogenic bacteria. The present study was aimed to isolate and identify the active compound of F-EtOH and compare its inhibitory activity against the biofilm of S. mutans as well as the cytotoxicity to oral fibroblast cells with F-EtOH. Compound isolation was done by column chromatography. The active compound was identified using liquid chromatography-mass spectrometry with electrospray ionization and nuclear magnetic resonance spectroscopy. It was found that the major compound of F-EtOH is brazilin. F-EtOH and brazilin were compared for inhibitory potential on the biofilms of three strains of S. mutans. The results exhibited that both F-EtOH and brazilin had potential on inhibiting biofilm formation and eradicating the preformed biofilms and their activity was dose dependent. F-EtOH showed significantly less toxic to normal periodontal ligament fibroblast than brazilin. At low concentration of 1- and 2-MBC, F-EtOH showed higher effective than brazilin. The results of our study suggest that the antibacterial activity of F-EtOH is according to the synergistic effects of the existing compounds including brazilin in F-EtOH.


Management of entecavir-resistant chronic hepatitis B with adefovir-based combination therapies.
To evaluate the long-term efficacy adefovir (ADV)-based combination therapies in entecavir (ETV)-resistant chronic hepatitis B (CHB) patients. Fifty CHB patients with genotypic resistance to ETV at 13 medical centers in South Korea were included for the analysis. All the patients received rescue therapy with the combination of ADV plus ETV (ADV/ETV, n = 23) or ADV plus lamivudine (LMV) (ADV/LMV, n = 27) for more than 12 mo. Patients were monitored at least every 3-4 mo during ADV-based combination therapy by clinical examination as well as biochemical and virological assessments. Hepatitis B virus (HBV) DNA levels were measured by real-time PCR and logarithmically transformed for analysis. Cumulative rates of virologic response (VR; HBV DNA < 20 IU/mL) were calculated using the Kaplan-Meier method, and the difference was determined by a log-rank test. Multivariate logistic regression and Cox proportional hazards models were used to identify independent risk factors significantly associated with short-term and long-term VR, respectively. Baseline median HBV DNA levels were 5.53 (2.81-7.63) log10 IU/mL. The most commonly observed ETV genotypic mutation sites were rt184 and rt202. Patients were treated for a median of 27 (12-45) mo. Overall, cumulative VR rates at 6, 12, 24, and 36 mo were 26%, 36%, 45%, and 68%, respectively. Patients treated with the ADV/ETV combination showed higher cumulative VR rates (35%, 43%, 65%, and 76%, respectively) than those with the ADV/LAM combination (18%, 30%, 30%, and 62%, respectively; P = 0.048). In the multivariate analysis, low baseline HBV DNA levels (< 5.2 log10 IU/mL) and initial virologic response at 3 mo (IVR-3; HBV DNA < 3.3 log10 IU/mL after 3 mo) were independent predictive factors for VR. Patients with favorable predictors achieved cumulative VR rates up to 90% at 36 mo. During the same period, the cumulative incidence of virologic breakthrough was as low as 6% in patients with the both favorable predictors. If tenofovir is not available, ADV/ETV combination could be considered in ETV-resistant patients with low HBV DNA titers, and may be continued if IVR-3 is achieved.


Polymorphisms in heterocyclic aromatic amines metabolism-related genes are associated with colorectal adenoma risk.
Colorectal adenoma (CRA) and colorectal cancer (CRC) risks have been linked to the intake of red and processed meat. Heterocyclic aromatic amines (HCA) formed herein during high temperature cooking, are metabolized by a variety of enzymes, and allelic variation in the coding genes could influence individual CRA risk. Associations of polymorphisms in NAT1, NAT2, GSTA1, SULT1A1, CYP1A2, UGT1A7, UGT1A9, GSTP1 genes with colorectal adenoma risk were investigated in a nested case-control study of the EPIC-Heidelberg cohort including 428 cases matched by age, sex and year of recruitment with one or two controls (n=828) with negative colonoscopy per case. Genoyping was preformed with the Sequenom MassArray system and the LightCycler 480. Conditional logistic regression was used to compute odds ratios (OR) and corresponding 95% confidence intervals (CI). For rs15561 (NAT1) and rs1057126 (NAT1), the rarer allel was significantly inversely associated with adenoma risk OR=0.80 (95% CI 0.65-0.97) and (OR=0.81 (95% CI 0.65-0.99) and, respectively). For the combined NAT2 alleles encoding for enzymes with medium (versus slow) activity we also observed a significantly inverse association with adenoma risk (OR=0.75; 95% CI 0.85-0.97). In addition, homozygous carriers of the A allele of rs3957357 (GSTA1), i.e., those with a decreased enzyme activity, had a decreased risk of colorectal adenoma with an OR of 0.68 (95% CI 0.50-0.92; AA versus GG/GA). Polymorphisms in the other tested genes did not modify the risk of colorectal adenomas. In conclusion, polymorphisms in NAT1, NAT2, and GSTA1 are related to colorectal adenoma risk in this German cohort.


Risk-reducing salpingo-oophorectomy for the prevention of BRCA1- and BRCA2-associated breast and gynecologic cancer: a multicenter, prospective study.
Risk-reducing salpingo-oophorectomy (RRSO) has been widely adopted as a key component of breast and gynecologic cancer risk-reduction for women with BRCA1 and BRCA2 mutations. Despite 17% to 39% of all BRCA mutation carriers having a mutation in BRCA2, no prospective study to date has evaluated the efficacy of RRSO for the prevention of breast and BRCA-associated gynecologic (ovarian, fallopian tube or primary peritoneal) cancer when BRCA2 mutation carriers are analyzed separately from BRCA1 mutation carriers. A total of 1,079 women 30 years of age and older with ovaries in situ and a deleterious BRCA1 or BRCA2 mutation were enrolled onto prospective follow-up studies at one of 11 centers from November 1, 1994 to December 1, 2004. Women self-selected RRSO or observation. Follow-up information through November 30, 2005, was collected by questionnaire and medical record review. The effect of RRSO on time to diagnosis of breast or BRCA-associated gynecologic cancer was analyzed using a Cox proportional-hazards model. During 3-year follow-up, RRSO was associated with an 85% reduction in BRCA1-associated gynecologic cancer risk (hazard ratio [HR] = 0.15; 95% CI, 0.04 to 0.56) and a 72% reduction in BRCA2-associated breast cancer risk (HR = 0.28; 95% CI, 0.08 to 0.92). While protection against BRCA1-associated breast cancer (HR = 0.61; 95% CI, 0.30 to 1.22) and BRCA2-associated gynecologic cancer (HR = 0.00; 95% CI, not estimable) was suggested, neither effect reached statistical significance. The protection conferred by RRSO against breast and gynecologic cancers may differ between carriers of BRCA1 and BRCA2 mutations. Further studies evaluating the efficacy of risk-reduction strategies in BRCA mutation carriers should stratify by the specific gene mutated.


Biocompatibility and biodegradability of metal organic frameworks for biomedical applications.
Metal organic frameworks (MOFs) are a unique class of smart hybrid materials that have recently attracted significant interest for catalysis, separation and biomedical applications. Different strategies have been developed to overcome the limitations of MOFs for bio-applications in order to produce a system with high biocompatibility and biodegradability. In this review, we outline the chemical and physical factors that dictate the biocompatibility and biodegradability characteristics of MOFs including the nature of the metal ions and organic ligands, size, surface properties and colloidal stability. This review includes the in vitro biodegradation and in vivo biodistribution studies of MOFs to better understand their pharmacokinetics, organ toxicity and immune response. Such studies can guide the design of future bio-friendly systems that bring us closer to safely translating these platforms into the pharmaceutical consumer market.


A pilot study to assess the efficacy of tariquidar to inhibit P-glycoprotein at the human blood-brain barrier with (R)-11C-verapamil and PET.
Tariquidar, a potent, nontoxic, third-generation P-glycoprotein (P-gp) inhibitor, is a possible reversal agent for central nervous system drug resistance. In animal studies, tariquidar has been shown to increase the delivery of P-gp substrates into the brain by severalfold. The aim of this study was to measure P-gp function at the human blood-brain barrier (BBB) after tariquidar administration using PET and the model P-gp substrate (R)-(11)C-verapamil. Five healthy volunteers underwent paired (R)-(11)C-verapamil PET scans and arterial blood sampling before and at 2 h 50 min after intravenous administration of tariquidar (2 mg/kg of body weight). The inhibition of P-gp on CD56-positive peripheral lymphocytes of each volunteer was determined by means of the (123)Rh efflux assay. Tariquidar concentrations in venous plasma were quantified using liquid chromatography/mass spectrometry. Tariquidar administration resulted in significant increases (Wilcoxon test for paired samples) in the distribution volume (DV, +24% +/- 15%) and influx rate constant (K(1), +49% +/- 36%) of (R)-(11)C-verapamil across the BBB (DV, 0.65 +/- 0.13 and 0.80 +/- 0.07, P = 0.043; K(1), 0.034 +/- 0.009 and 0.049 +/- 0.009, P = 0.043, before and after tariquidar administration, respectively). A strong correlation was observed between the change in brain DV after administration of tariquidar and tariquidar exposure in plasma (r = 0.90, P = 0.037). The mean plasma concentration of tariquidar achieved during the second PET scan (490 +/- 166 ng/mL) corresponded to 100% inhibition of P-gp function in peripheral lymphocytes. Tariquidar significantly increased brain penetration of (R)-(11)C-verapamil-derived activity due to increased influx. As opposed to peripheral P-gp function, central P-gp inhibition appeared to be far from complete after the administered tariquidar dose.


Discovery of the presence and functional expression of cannabinoid CB2 receptors in brain.
Two well-characterized cannabinoid receptors (CBrs), CB1 and CB2, mediate the effects of cannabinoids and marijuana use, with functional evidence for other CBrs. CB1 receptors are expressed primarily in brain and peripheral tissues. For over a decade several laboratories were unable to detect CB2 receptors in brain and were known to be intensely expressed in peripheral and immune tissues and have traditionally been referred to as peripheral CB2 CBrs. We have reported the discovery and functional presence of CB2 cannabinoid receptors in mammalian brain that may be involved in depression and drug abuse and this was supported by reports of identification of neuronal CB2 receptors that are involved in emesis. We used RT-PCR, immunoblotting, hippocampal cultures, immunohistochemistry, transmission electron microscopy, and stereotaxic techniques with behavioral assays to determine the functional expression of CB2 CBrs in rat brain and mice brain exposed to chronic mild stress (CMS) or those treated with abused drugs. RT-PCR analyses supported the expression of brain CB2 receptor transcripts at levels much lower than those of CB1 receptors. In situ hybridization revealed CB2 mRNA in cerebellar neurons of wild-type but not of CB2 knockout mice. Abundant CB2 receptor immunoreactivity (iCB2) in neuronal and glial processes was detected in brain and CB2 expression was detected in neuron-specific enolase (NSE) positive hippocampal cell cultures. The effect of direct CB2 antisense oligonucleotide injection into the brain and treatment with JWH015 in motor function and plus-maze tests also demonstrated the functional presence of CB2 cannabinoid receptors in the central nervous system (CNS). Thus, contrary to the prevailing view that CB2 CBrs are restricted to peripheral tissues and predominantly in immune cells, we demonstrated that CB2 CBrs and their gene transcripts are widely distributed in the brain. This multifocal expression of CB2 immunoreactivity in brain suggests that CB2 receptors may play broader roles in the brain than previously anticipated and may be exploited as new targets in the treatment of depression and substance abuse.


Widespread distribution of binding sites for the novel Ca2+-mobilizing messenger, nicotinic acid adenine dinucleotide phosphate, in the brain.
Nicotinic acid adenine dinucleotide phosphate (NAADP) is a potent Ca(2+)-mobilizing agent in invertebrate eggs that has recently been shown to be active in certain mammalian and plant systems. Little, however, is known concerning the properties of putative NAADP receptors. Here, for the first time, we report binding sites for NAADP in brain. In contrast to sea urchin egg homogenates, [(32)P]NAADP bound reversibly to multiple sites in brain membranes. The rank order of potency of NAADP, 2',3'-cyclic NAADP and 3'-NAADP in displacing [(32)P]NAADP was, however, the same in the two systems and in agreement with their ability to mobilize Ca(2+) from homogenates. These data indicate that [(32)P]NAADP likely binds to receptors mediating Ca(2+) mobilization. Autoradiography revealed striking heterogeneity in the distribution of [(32)P]NAADP binding sites throughout the brain. Our data strongly support a role for NAADP-induced Ca(2+) signaling in the brain.


What Is the Diagnostic Accuracy of the Duck Walk Test in Detecting Meniscal Tears?
Clinical weightbearing provocation tests, like the duck walk test, may be of value in diagnosing or screening for medial meniscal tears. However, evidence of the diagnostic accuracy of the duck walk test is lacking. (1) To determine the sensitivity and specificity of the duck walk test in diagnosing medial meniscal tears. (2) To determine whether tear location, tear cause (traumatic versus degenerative), and ACL insufficiency were associated with differences in the sensitivity and specificity of the test. A convenience sample of 136 patients of all ages was retrospectively analyzed by evaluating the outpatient knee clinic appointment list of one orthopaedic surgeon for patients with a broad range of knee injuries who had a prior MRI before (24%) or after (76%) physical examination and had a duck walk test stated in their patient records. Of 230 patients with MRI requested by one orthopaedic surgeon attributable to knee complaints, 136 (59%) fulfilled the inclusion criteria; 70 (52%) patients were male and 66 (49%) were female, with a mean age of 42 (± SD 14) years. The duck walk test was performed in case of suspected meniscal injury, based on mechanism of injury, general joint line pain, and/or mechanical complaints (ie, locking, giving away). The test is performed by squatting and "waddling" before rising and is positive in case of general joint line pain or painful "clicking". Interobserver repeatability was not evaluated, but the test is well defined and leaves little room for difference in interpretation. Diagnostic accuracy measures were evaluated. Since the convenience sample in this study consisted of patients who had a duck walk test and MRI, and a positive result of the duck walk test almost certainly increased the probability that MRI would be ordered in the majority (76%) of the patients, the test properties calculated here-especially sensitivity-should be considered inflated. The calculated sensitivity of the duck walk test was 71% (95% CI, 59%-81%) and there was low specificity of 39% (95% CI, 27%-52%). We found no difference in sensitivity between medial (67%; 95% CI, 51%-80%) and lateral (76%; 95% CI, 50%-92%; p = 0.492) meniscal tears. With the numbers available, we compared these patients with patients without a history of trauma and with an intact ACL. We found no difference among patients with traumatic tears (79%; 95% CI, 59%-91%; p = 0.253) and in patients with ACL tears (77%; 95% CI, 46%-94%; p = 0.742). Because of the issue of verification bias, the actual sensitivity of this test in practice is likely much lower than the calculated sensitivity we observed. In addition, the test did not seem to perform better in patients with trauma or ACL insufficiency, nor was it more effective in detecting medial than lateral tears, although the numbers on some of those comparisons were rather small. Based on these results, we conclude that used alone, the duck walk test likely has little value in practice as a screening test. However, it is conceivable that it could be used in combination with other provocative tests for screening purposes. Future studies might consider using it as a means to best identify which patients should undergo MRI for the possibility of a meniscal tear. Level III, diagnostic study.


Re-visioning Academic Medicine Through a Constructionist Lens.
Constructionism in academic medicine matters. It encourages educators and researchers to question taken-for-granted assumptions, paying close attention to socially and historically contingent meanings. In this Invited Commentary, the authors explain what constructionism is; examine its ontological, epistemological, and axiological underpinnings; and outline its common methodologies and methods. Although constructivism favors the individual, constructionism privileges the social as the controlling force behind the construction of meaning. Where micro-constructionism attends to the minutiae of language, macro-constructionism focuses on broader discourses reproduced through material and social practices and structures. While social constructionists might situate themselves at any point on the relativist-realist continuum, many constructionists focus on constructionism as epistemology (the nature of knowledge) rather than ontology (the nature of reality). From an epistemological standpoint, constructionism asserts that how we come to know the world is constructed through social interaction. Constructionism thus values language, dialogue, and context, in addition to internal coherence between epistemology, methodology, and methods. Constructionism similarly values the concepts of dependability, authenticity, credibility, confirmability, reflexivity, and transferability. It also embraces the researcher-researched relationship. Given the privileging of language, qualitative methodologies and methods are key in constructionism, with constructionist-type questions focusing on how people speak. Here, the authors encourage the reader to develop an understanding of constructionism to re-vision academic medicine through a constructionist lens.


Gamma-hydroxybutyrate withdrawal syndrome.
Gamma-hydroxybutyrate (GHB) withdrawal syndrome is increasingly encountered in emergency departments among patients presenting for health care after discontinuing frequent GHB use. This report describes the characteristics, course, and symptoms of this syndrome. A retrospective review of poison center records identified 7 consecutive cases in which patients reporting excessive GHB use were admitted for symptoms consistent with a sedative withdrawal syndrome. One additional case identified by a medical examiner was brought to our attention. These medical records were reviewed extracting demographic information, reason for presentation and use, concurrent drug use, toxicology screenings, and the onset and duration of clinical signs and symptoms. Eight patients had a prolonged withdrawal course after discontinuing chronic use of GHB. All patients in this series were psychotic and severely agitated, requiring physical restraint and sedation. Cardiovascular effects included mild tachycardia and hypertension. Neurologic effects of prolonged delirium with auditory and visual hallucinations became episodic as the syndrome waned. Diaphoresis, nausea, and vomiting occurred less frequently. The onset of withdrawal symptoms in these patients was rapid (1 to 6 hours after the last dose) and symptoms were prolonged (5 to 15 days). One death occurred on hospital day 13 as withdrawal symptoms were resolving. In our patients, severe GHB dependence followed frequent ingestion every 1 to 3 hours around-the-clock. The withdrawal syndrome was accompanied initially by symptoms of anxiety, insomnia, and tremor that developed soon after GHB discontinuation. These initial symptoms may progress to severe delirium with autonomic instability.


Negotiating access to the Nigerian healthcare system: the experiences of HIV-positive men who have sex with men.
Men who have sex with men in sub-Saharan Africa continue to have a disproportionately higher burden of HIV than their counterparts in the general population. In this qualitative study, barriers to participants accessing healthcare services in Nigeria, a country where same-sex relationships are criminalised and considered societally abhorrent, were explored. Four focus group discussions and 21 semi-structured interviews were conducted with HIV-positive men who have sex with men recruited from 3 NGOs in Abuja and Lagos, Nigeria in 2016. Interviews were transcribed verbatim and analysed thematically using NVivo. The findings of this study revealed the numerous barriers these men encounter accessing general (government and private) healthcare facilities. Data were organised into two categories: barriers associated with the wider legal context in relation to HIV-positive men who have sex with men and those with service delivery. There were barriers reported in relation to accessing an emerging service, which had hitherto sought to address some barriers encountered in general healthcare facilities. Findings point to the importance of facilitating a more enabling social and political environment whereby men who have sex with men can freely access healthcare services, potentially through these facilities.


Minimal impact of response shift for SF-12 mental and physical health status in homeless and vulnerably housed individuals: an item-level multi-group analysis.
The purpose of this study was to examine whether homeless or vulnerably housed individuals experienced response shift over a 12-month time period in their self-reported physical and mental health status. Data were obtained from the Health and Housing in Transition study, a longitudinal multi-site cohort study in Canada (N = 1190 at baseline). Multi-group confirmatory factor analysis (MG-CFA) and methods for response shift detection at the item level, based on the approach by Oort, were used to test for reconceptualization, reprioritization, and recalibration response shift on the SF-12 in four groups of individuals who were homeless (n = 170), housed (n = 437), or who reported a change in their housing status [from homeless to housed (n = 285) or housed to homeless (n = 73)] over a 12-month time period. Mean and variance adjusted weighted-least squares estimation was used to accommodate the ordinal and binary distributions of the SF-12 items. Using MG-CFA, a strict invariance model showed that the measurement model was equivalent for the four groups at baseline. Although we found small but statistically significant response shift for several measurement model parameters, the impact on the predicted average mental and physical health scores within each of the groups was small. Response shift does not appear to be a significant concern when using the SF-12 to obtain change scores over a 12-month period in this population.


Managing the stresses of group-living in the transition to village life.
Group living is stressful for all mammals, and these stresses limit the size of their social groups. Humans live in very large groups by mammal standards, so how have they solved this problem? I use homicide rates as an index of within-community stress for humans living in small-scale ethnographic societies, and show that the frequency of homicide increases linearly with living-group size in hunter-gatherers. This is not, however, the case for cultivators living in permanent settlements, where there appears to be a 'glass ceiling' below which homicide rates oscillate. This glass ceiling correlates with the adoption of social institutions that allow tensions to be managed. The results suggest (a) that the transition to a settled lifestyle in the Neolithic may have been more challenging than is usually assumed and (b) that the increases in settlement size that followed the first villages necessitated the introduction of a series of social institutions designed to manage within-community discord.


Variation in levels of anxiety to dental treatment among nonorphan and orphan children living under different systems.
It is essential to understand the factors influencing the level of anxiety to dental treatment among different children as it can influence seeking dental care. Here, we assessed the impact of parental loss on dental anxiety among 6-13-year-old children. A total of 444 children within the age group 6-13 years were selected. Group 1 consisted of orphan children living in government-run orphanages, Group 2 consisted of orphan children taken care by a person with a motherly relationship, Group 3 consisted of abandoned children living in private organization and Group 4 consisted of children living with their parents. Dental anxiety was measured using children's fear survey schedule-dental subscale and modified faces version of modified child dental anxiety scale. The highest number of anxious children were observed in Group 4 and the difference in the anxiety levels among the four groups was found to be highly statistically significant. Children living in government-run orphanages had least dental anxiety. All the orphans may not have the same anxiety levels and the environment of upbringing the orphans plays a significant role in the development of the anxiety.


Towards a molecular definition of worker sterility: differential gene expression and reproductive plasticity in honey bees.
We show that differences in the reproductive development of honey bee workers are associated with locus-specific changes to abundance of messenger RNA. Using a cross-fostering field experiment to control for differences related to age and environment, we compared the gene expression profiles of functionally sterile workers (wild-type) and those from a mutant strain in which workers are reproductively active (anarchist). Among the set of three genes that are significantly differentially expressed are two major royal jelly proteins that are up-regulated in wild-type heads. This discovery is consistent with sterile workers synthesizing royal jelly as food for developing brood. Likewise, the relative underexpression of these two royal jellies in anarchist workers is consistent with these workers' characteristic avoidance of alloparental behaviour, in favour of selfish egg-laying. Overall, there is a trend for the most differentially expressed genes to be up-regulated in wild-type workers. This pattern suggests that functional sterility in honey bee workers may generally involve the expression of a suite of genes that effectively 'switch' ovaries off, and that selfish reproduction in honey bee workers, though rare, is the default developmental pathway that results when ovary activation is not suppressed.


Adiponectin, type 2 diabetes and the metabolic syndrome: lessons from human genetic studies.
Adiponectin, a protein exclusively secreted by adipose tissue but present at low levels in obesity, is now widely recognised as a key determinant of insulin sensitivity and of protection against obesity-associated metabolic syndrome. In this review we explain how genetic findings have contributed to a better understanding of the physiological role of adiponectin in humans. The adiponectin-encoding gene, ADIPOQ (ACDC), is very polymorphic: many frequent exonic synonymous, intronic and promoter single-nucleotide polymorphisms (SNPs) have been identified, as well as a few rare exonic amino acid substitutions. Several of these variations additively contribute to the modulation of adiponectin level and function, and associate with insulin sensitivity, type 2 diabetes and vascular complications of obesity.


The use of digital photographs for the diagnosis of hand osteoarthritis: the AGES-Reykjavik study.
The objective of the study was to standardize a method using digital photographs to diagnose and grade hand osteoarthritis (HOA), to compare it with radiographs and clinical examination with regard to prevalence and relation to symptoms, and finally to construct a simple shortened version suitable for use in very large studies, where a global estimate may be preferable. High quality photographs with standard distance and hand positioning were analysed for the presence of HOA and subsequently compared with standard radiographs and clinical examination in 381 random participants in the AGES-Reykjavik Study, a large population study. The mean age of the participants was 76 years. Using the photographic method, the most commonly affected joints were the second DIP joints followed by the third DIP joints and second and third PIP joints. Both interobserver (ICC = 0.83) and intraobserver reading agreements (ICC = 0.89) were acceptable. On comparison with radiography and clinical examination, aggregate scores were significantly correlated (R(s) 0.35-0.69), more so in females (R(s) 0.53-0.72) than males. Hand pain in males showed very little association with HOA findings by the three methods but all methods showed a comparable moderate association with hand pain in females. The performance of photography in predicting pain on most days for at least a month in females was comparable to that of radiography and clinical examination (AUC 0.63 p = 0.004). Analysis of intermittent pain yielded similar results for in the DIP and PIP joints (OR 3.2-3.3, p < 0.01), but for the CMC1 joints, both radiography (OR 9.0, p < 0.0001), and clinical examination (OR 9.8, p < 0.0001), had higher predictive odds ratios for pain than photography (OR 3.6, p < 0.0001)., A shortened, rapidly performed form of reading photographs also showed a high degree of correlation with the other methods (R(s) 0.56-0.82). High quality hand photographs can be used to diagnose and grade hand osteoarthritis. The method has the advantage of being inexpensive and easy to perform. By using a slightly simplified method of reading, it appears to be highly suitable for use in large studies.


The clinical course of chronic periodontitis.
The purpose of this study was to assess the long-term influence of gingival inflammation on tooth loss. The data originated from a 26-year longitudinal study of Norwegian males, who practiced adequate daily oral home care and received "state-of-the-art" dental care. The initial examination in 1969 included 565 individuals aged between 16 and 34 years. Subsequent examinations took place in 1971, 1973, 1975, 1981, 1988 and 1995. Thus, the study covers the age range of 16-59 years. The teeth were divided into three tooth groups (I-III) reflecting the history of inflammation of the surrounding gingiva (gingival index (GI) scores) over 26 years: (I) teeth with surrounding gingival units scoring a minimum of one site with GI=0 and a maximum of three sites with GI=1, (II) teeth with surrounding gingival units scoring a minimum of one site with GI=1 and a maximum of three sites with GI=2 over the observation periods and (III) teeth with surrounding gingival units always scoring a minimum of GI=2 (bleeding on probing) at all sites over the observation period. At baseline (1969), out of possible 15,820 teeth (565 x 28), 15,383 teeth were present. Four hundred and thirty-seven teeth had already been missing for unknown reasons. By 1995, 13,159 teeth were reexamined, i.e. over the 26-year observation period only 126 (0.95%) teeth were lost. Only 16 (0.28%) of 5793 teeth belonging to GI-Severity Group I were lost. In the GI-Severity Group II, however, 78 (2.28%) out of 3348 teeth were lost, and 13 (11.21%) of 103 teeth with GI-Severity Group III were lost. Teeth with GI-Severity Group III yielded an odds ratio for tooth loss that was 46 times higher than that of teeth with GI-Severity Group I, and five times higher than that of teeth with GI-Severity Group II over 26 years. Furthermore, teeth with the GI-Severity Group II had a nine times higher risk for tooth loss than teeth with the GI-Severity Group I. The GI-Severity Group I retained 99.5% of the teeth after a tooth age of 51 years. The GI-Severity Group II retained 93.8% of the teeth after a tooth age of 50 years. However, in the GI-Severity Group III, 63.4% of the teeth were retained for a tooth age of 47 years. Teeth surrounded with inflammation-free gingival tissues were maintained for a tooth age of 51 years, while teeth consistently surrounded with inflamed gingivae yielded a 46-times higher risk to be lost. Only two-thirds of such teeth were maintained throughout the 26-year observation period. This documents the role of gingival inflammation as a risk factor for future tooth loss.


Sc3p hydrophobin organization in aqueous media and assembly onto surfaces as mediated by the associated polysaccharide schizophyllan.
Sc3p is the major hydrophobin produced and excreted by the wood-rotting fungus Schizophyllum commune when grown as a monokaryon in culture. In addition to Sc3p, several lower molecular weight proteins of unknown function and a high molecular weight polysaccharide, schizophyllan, are present in this fungal culture medium. Previous studies in our laboratory indicated that schizophyllan and hydrophobin, as produced in the culture supernatant, work in concert to ultimately produce stable films on hydrophilic substrates. We report a new procedure for isolation of hydrophobin that does not require treatment with strong organic acids. This procedure allows a systematic evaluation of the importance of the individual hydrophobin and schizophyllan components in solution stabilization and film formation from a conformational state more representative of that found during fungal production. Experimental results indicate that schizophyllan stabilizes small hydrophobin multimers in aqueous solution. In addition, schizophyllan is necessary for subsequent hydrophobin assembly into films on hydrophilic surfaces. By contrast, hydrophobin alone is sufficient for thin film assembly onto a hydrophobic surface; however, hydrophobin aggregation at high protein concentration in the absence of schizophyllan may limit the efficiency of film formation. A model has been proposed in which the polysaccharide acts as a hydrophilic stabilizer of hydrophobin in solution preventing self-aggregation. The schizophyllan-stabilized complex may also allow conformational mobility for lateral reorganization during deposition and film coalescence.


Some aspects of astroglial functions and aluminum implications for neurodegeneration.
The present decade had witnessed an unprecedented attention focused on glial cells as a result of their unusual physiological roles that are being unraveled. It is now known that, rather than being a mere supporter of neurons, astroglia are actively involved in their modulation. The aluminum hypothesis seems to have been laid to rest, probably due to contradictory epidemiological reports on it as a causative factor of neurodegenerative diseases. Surprisingly, newer scientific evidences continue to appear and recent findings have implicated astrocytes as the principal target of its toxic action. In view of the likely detrimental effects of the interaction between these two infamous partners in neuroscience on neurons and nervous system, we have reviewed some aspects of glia-neuron interaction and discussed the implications of aluminum-impaired astrocytic functions on neurodegeneration. Because sporadic causes still account for the majority of the neurodegenerative diseases of which Alzheimer's disease is the most prominent, it has been suggested that neurotoxicologists should not relent in screening for the environmental agents, such as aluminum, and that considerable attention should be given to glial cells in view of the likely implications of environmental toxicants on their never-imagined newly reported roles in the central nervous system (CNS).


Wound with Diabetes: Present Scenario and Future.
Diabetes is a chronic metabolic disorder of the endocrine system characterized by an increase in blood glucose level. Several factors, such as pancreatic damage, oxidative stress, infection, genetic factor, obesity, liver dysfunction, play a vital role in the pathogenesis of diabetes, which further leads to serious diabetic complications. The diabetic wound is one such complication where the wound formation occurs, especially due to pressure and its healing process is disrupted due to factors, such as hyperglycemia, neuropathy, nephropathy, peripheral vascular disease, reduction of blood flow, atherosclerosis, impaired fibroblast. The process of wound healing is delayed due to different abnormalities like alteration in nitric oxide level, increase in aldose reductase, sorbitol, and fructose. Therefore, diabetic wound requires more time to heal as compared to the normal wound. Healing time is delayed in diabetic wound due to many factors, such as stress, decreased oxygenation supply, infection, decreased blood flow, impaired proliferation and migration rate, impaired growth factor production, impaired keratinocytes proliferation and migration, and altered vascular endothelial mediators. The current treatment for diabetic wounds includes wound patches, oxygenation therapy, hydrogel patches, gene therapy, laser therapy, and stem cell therapy. Medications with phytoconstituents are also one way to manage the diabetic wound, but it is not more effective for quick healing. The objective of this review is to understand the potential of various management options which are available for diabetic wound, with a special focus on biological cells.


Monitoring and identification of sepsis development through a composite measure of heart rate variability.
Tracking the physiological conditions of a patient developing infection is of utmost importance to provide optimal care at an early stage. This work presents a procedure to integrate multiple measures of heart rate variability into a unique measure for the tracking of sepsis development. An early warning system is used to illustrate its potential clinical value. The study involved 17 adults (age median 51 (interquartile range 46-62)) who experienced a period of neutropenia following chemoradiotherapy and bone marrow transplant; 14 developed sepsis, and 3 did not. A comprehensive panel (N = 92) of variability measures was calculated for 5 min-windows throughout the period of monitoring (12 ± 4 days). Variability measures underwent filtering and two steps of data reduction with the objective of enhancing the information related to the greatest degree of change. The proposed composite measure was capable of tracking the development of sepsis in 12 out of 14 patients. Simulating a real-time monitoring setting, the sum of the energy over the very low frequency range of the composite measure was used to classify the probability of developing sepsis. The composite revealed information about the onset of sepsis about 60 hours (median value) before of sepsis diagnosis. In a real monitoring setting this quicker detection time would be associated to increased efficacy in the treatment of sepsis, therefore highlighting the potential clinical utility of a composite measure of variability.


Interval hypoxic training improves autonomic cardiovascular and respiratory control in patients with mild chronic obstructive pulmonary disease.
Chronic obstructive pulmonary disease (COPD) is associated with cardiac autonomic nervous system dysregulation. This study evaluates the effects of interval hypoxic training on cardiovascular and respiratory control in patients with mild COPD. In 18 eucapnic normoxic mild COPD patients (age 51.7 +/- 2.4 years, mean +/- SEM), randomly assigned to either training or placebo group, and 14 age-matched healthy controls (47.7 +/- 2.8 years), we monitored end-tidal carbon dioxide, airway flow, arterial oxygen saturation, electrocardiogram, and continuous noninvasive blood pressure at rest, during progressive hypercapnic hyperoxia and isocapnic hypoxia to compare baroreflex sensitivity to hypoxia and hypercapnia before and after 3 weeks of hypoxic training. In double-blind fashion, both groups received 15 sessions of passive intermittent hypoxia (training group) or normoxia (placebo group). For the hypoxia group, each session consisted of three to five hypoxic (15-12% oxygen) periods (3-5 min) with 3-min normoxic intervals. The placebo group inhaled normoxic air. Before training, COPD patients showed depressed baroreflex sensitivity, as compared with healthy individuals, without evident chemoreflex abnormalities. After training, in contrast to placebo group, the training group showed increased (P < 0.05) baroreflex sensitivity up to normal levels and selectively increased hypercapnic ventilatory response (P < 0.05), without changes in hypoxic ventilatory response. Eucapnic normoxic mild COPD patients already showed signs of cardiovascular autonomic abnormalities at baseline, which normalized with hypoxic training. If confirmed in more severe patients, interval hypoxic training may be a therapeutic strategy to rebalance early autonomic dysfunction in COPD patients.


Recurrence of depression after bereavement-related depression: evidence for the validity of DSM-IV bereavement exclusion from the Epidemiologic Catchment Area Study.
The DSM-IV diagnostic criteria for major depressive disorder exclude bereavement-related depressive episodes that are brief and lack certain severe symptoms and are thus better explained as normal grief responses. However, the DSM-5 Task Force proposes to eliminate this exclusion because of a lack of evidence that such episodes differ relevantly from standard major depression. Using the two-wave longitudinal Epidemiologic Catchment Area Study, we compared 1-yr depression recurrence rates at wave 2 of four groups at wave 1 baseline: (1) those with no history of depressive disorder (n = 18,239), (2) those who had only lifetime excludable bereavement-related depression (n = 25), (3) those with brief-episode (≤ 2 months duration) lifetime standard depressive disorder (n = 446), and (4) those with nonbrief lifetime standard depressive disorder (n = 581). The recurrence rate in the excludable-depression group (3.7%) was not significantly different from the no-history group (1.7%) but was significantly and substantially lower than in the brief and nonbrief standard depression groups (14.4% and 16.2%, respectively). These findings confirm findings reported by Mojtabai (Arch Gen Psychiatry 68:920-928, 2011) using a different data set and time frame and thus substantially strengthen the support for the validity of bereavement exclusion and for its preservation in the DSM-5.


C-reactive protein is protective against Streptococcus pneumoniae infection in mice.
C-reactive protein (CRP) has several properties that suggest that it may function as a bacterial opsonin. CRP shows binding reactivity with pneumococcal C-polysaccharide, the cell wall carbohydrate of Streptococcus pneumoniae. In this study we have demonstrated protection of mice against serotypes 3 and 4 of S. pneumoniae infection by a single prior injection of CRP. This effect was seen both in mice that lacked antibody to phosphocholine and in normal mice. Thus the opsonic properties of CRP previously described may be related to protection against pneumococcal infection.


Record linked retrospective cohort study of 4.6 million people exploring ethnic variations in disease: myocardial infarction in South Asians.
Law and policy in several countries require health services to demonstrate that they are promoting racial/ethnic equality. However, suitable and accurate data are usually not available. We demonstrated, using acute myocardial infarction, that linkage techniques can be ethical and potentially useful for this purpose. The linkage was based on probability matching. Encryption of a unique national health identifier (the Community Health Index (CHI)) ensured that information about health status and census-based ethnicity could not be ascribed to an identified individual. We linked information on individual ethnic group from the 2001 Census to Scottish hospital discharge and mortality data. Overall, 94% of the 4.9 million census records were matched to a CHI record with an estimated false positive rate of less than 0.1 %, with 84.9 - 87.6% of South Asians being successfully linked. Between April 2001 and December 2003 there were 126 first episodes of acute myocardial infarction (AMI) among South Asians and 30,978 among non-South Asians. The incidence rate ratio was 1.45 (95% CI 1.17, 1.78) for South Asian compared to non-South Asian men and 1.80 (95% CI 1.31, 2.48) for South Asian women. After adjustment for age, sex and any previous admission for diabetes the hazard ratio for death following AMI was 0.59 (95% CI 0.43, 0.81), reflecting better survival among South Asians. The technique met ethical, professional and legal concerns about the linkage of census and health data and is transferable internationally wherever the census (or population register) contains ethnic group or race data. The outcome is a retrospective cohort study. Our results point to increased incidence rather than increased case fatality in explaining high CHD mortality rate. The findings open up new methods for researchers and health planners.


Studies on the resting behaviour and host choice of Anopheles gambiae and An. arabiensis from Muleba, Tanzania.
The relative efficacy of a mechanical (Prokopack) collection method vs. manual aspiration in the collection of resting mosquitoes was evaluated in northern Tanzania before and after an intervention using indoor residual spraying and longlasting insecticide-treated nets. In smoke-free houses mosquitoes were collected from the roof and walls, but in smoky houses mosquitoes were found predominantly on the walls. Anopheles gambiae (Diptera: Culicidae) constituted 97.7% of the 312 An. gambiae complex specimens identified before but only 19.3% of the 183 identified after the intervention. A single sampling with the Prokopack collected a third of the available insects. Anopheles gambiae completed its gonotrophic development indoors, whereas Anopheles arabiensis did so outdoors. In both species gonotrophic development took 2 days. Most unfed resting An. arabiensis collected outdoors were virgins, whereas the majority of engorged insects were parous (with well-contracted sacs). Daily survival was estimated to be 80.0%. Only 9.4% of the engorged An. arabiensis collected outdoors and 47.1% of those collected indoors had fed on humans. Using the Prokopack sampler is more efficient than manual methods for the collection of resting mosquitoes. Malaria transmission may have been affected by a change in vector composition resulting from a change in feeding, rather than reduced survival. Monitoring the proportions of members of the An. gambiae complex may provide signals of an impending breakdown in control.


Conceptualization and measurement of criminal thinking: initial validation of the Criminogenic Thinking Profile.
This article describes two studies concerning the development of a new measure of criminal thinking, the CriminogenicThinking Profile (CTP), influenced by the construct of psychopathy, and traditional models of cognitive-behavioral therapy (CBT). An experimental item pool based on verbalizations from offenders served as the pilot version of the instrument. Principal components analysis of the items resulted in a 62-item, eight-factor scale that was internally consistent. In terms of content, six of the resulting factors were conceptually related to psychopathy, one to CBT, and one to neutralization theory. The factor structure and internal reliability was supported by a subsequent confirmatory factor analysis. Initial support for the CTP's convergent validity was indicated by its positive correlations with psychopathy and personality disorders associated with criminal, aggressive, and impulsive behaviors. The CTP's divergent validity was supported by its inverse correlations with indices of healthy personality functioning. The CTP offers a somewhat different constellation of thinking patterns than those found on previously published criminal thinking instruments. The utility of the CTP to identify relevant cognitive targets for offender treatment is a recommended area of future research.


Microglial control of astrocytes in response to microbial metabolites.
Microglia and astrocytes modulate inflammation and neurodegeneration in the central nervous system (CNS)1-3. Microglia modulate pro-inflammatory and neurotoxic activities in astrocytes, but the mechanisms involved are not completely understood4,5. Here we report that TGFα and VEGF-B produced by microglia regulate the pathogenic activities of astrocytes in the experimental autoimmune encephalomyelitis (EAE) mouse model of multiple sclerosis. Microglia-derived TGFα acts via the ErbB1 receptor in astrocytes to limit their pathogenic activities and EAE development. Conversely, microglial VEGF-B triggers FLT-1 signalling in astrocytes and worsens EAE. VEGF-B and TGFα also participate in the microglial control of human astrocytes. Furthermore, expression of TGFα and VEGF-B in CD14+ cells correlates with the multiple sclerosis lesion stage. Finally, metabolites of dietary tryptophan produced by the commensal flora control microglial activation and TGFα and VEGF-B production, modulating the transcriptional program of astrocytes and CNS inflammation through a mechanism mediated by the aryl hydrocarbon receptor. In summary, we identified positive and negative regulators that mediate the microglial control of astrocytes. Moreover, these findings define a pathway through which microbial metabolites limit pathogenic activities of microglia and astrocytes, and suppress CNS inflammation. This pathway may guide new therapies for multiple sclerosis and other neurological disorders.


The nucleotide switch in Cdc42 modulates coupling between the GTPase-binding and allosteric equilibria of Wiskott-Aldrich syndrome protein.
The GTP/GDP nucleotide switch in Ras superfamily GTPases generally involves differential affinity toward downstream effectors, with the GTP-bound state having a higher affinity for effector than the GDP-bound state. We have developed a quantitative model of allosteric regulation of the Wiskott-Aldrich syndrome protein (WASP) by the Rho GTPase Cdc42 to better understand how GTPase binding is coupled to effector activation. The model accurately predicts WASP affinity for Cdc42, activity toward Arp2/3 complex, and activation by Cdc42 as functions of a two-state allosteric equilibrium in WASP. The ratio of GTPase affinities for the inactive and active states of WASP is appreciably larger for Cdc42-GTP than for Cdc42-GDP. The greater ability to distinguish between the two states of WASP makes Cdc42-GTP a full WASP agonist, whereas Cdc42-GDP is only a partial agonist. Thus, the nucleotide switch controls not only the affinity of Cdc42 for its effector but also the efficiency of coupling between the Cdc42-binding and allosteric equilibria in WASP. This effect can ensure high fidelity and specificity in Cdc42 signaling in crowded membrane environments.


Accidents in a psychiatric hospital.
In a six-month study of psychiatric hospital in-patients, it was found that accidents more often occurred among females and among patients on psychogeriatric wards. Falls were the most common form of accident. Accidents resulting from the aggressive behaviour of other patients usually occurred on long-stay and rehabilitation wards, with middle-aged patients the victims.


Validity and reliability of the 'Ten Questions' questionnaire for detecting moderate to severe neurological impairment in children aged 6-9 years in rural Kenya.
The 'Ten Questions' Questionnaire (TQQ) is used to detect severe neurological impairment in children living in resource-poor countries. Its usefulness has been established in Asia and the Caribbean, but there are a few published studies from Africa. We evaluated the TQQ as part of a larger study of neurological impairment in a rural community, on the coast of Kenya. The study was conducted in two phases from June 2001 to May 2002; in phase one, a community household screening of 10,218 children aged 6-9 years using the TQQ was performed. Phase two involved a comprehensive clinical and psychological assessment of all children testing positive on the TQQ (n = 810) and an equivalent number of those testing negative (n = 766). Data were interpreted using the impairment-specific approach. Overall, the sensitivity rates for screening the different impairments were: cognitive (70.0%), motor (71.4%), epilepsy (100%), hearing (87.4%) and visual (77.8%). All the specificity rates were greater than 96%. However, the positive predictive values were low, and ranged from 11 to 33%. These results are similar to those from other continents and provide evidence that the TQQ can be used to compare the epidemiology of moderate/severe impairment in different parts of the world. Furthermore, the TQQ can be used to screen for moderately/severely impaired children in resource-poor countries; however, the low positive predictive values mean that other assessments are required for confirmation.


Ablation of TFR1 in Purkinje Cells Inhibits mGlu1 Trafficking and Impairs Motor Coordination, But Not Autistic-Like Behaviors.
Group 1 metabotropic glutamate receptors (mGlu1/5s) are critical to synapse formation and participate in synaptic LTP and LTD in the brain. mGlu1/5 signaling alterations have been documented in cognitive impairment, neurodegenerative disorders, and psychiatric diseases, but underlying mechanisms for its modulation are not clear. Here, we report that transferrin receptor 1 (TFR1), a transmembrane protein of the clathrin complex, modulates the trafficking of mGlu1 in cerebellar Purkinje cells (PCs) from male mice. We show that conditional knock-out of TFR1 in PCs does not affect the cytoarchitecture of PCs, but reduces mGlu1 expression at synapses. This regulation by TFR1 acts in concert with that by Rab8 and Rab11, which modulate the internalization and recycling of mGlu1, respectively. TFR1 can bind to Rab proteins and facilitate their expression at synapses. PC ablation of TFR1 inhibits parallel fiber-PC LTD, whereas parallel fiber-LTP and PC intrinsic excitability are not affected. Finally, we demonstrate that PC ablation of TFR1 impairs motor coordination, but does not affect social behaviors in mice. Together, these findings underscore the importance of TFR1 in regulating mGlu1 trafficking and suggest that mGlu1- and mGlu1-dependent parallel fiber-LTD are associated with regulation of motor coordination, but not autistic behaviors.SIGNIFICANCE STATEMENT Group 1 metabotropic glutamate receptor (mGlu1/5) signaling alterations have been documented in cognitive impairment, neurodegenerative disorders, and psychiatric diseases. Recent work suggests that altered mGlu1 signaling in Purkinje cells (PCs) may be involved in not only motor learning, but also autistic-like behaviors. We find that conditional knock-out of transferrin receptor 1 (TFR1) in PCs reduces synaptic mGlu1 by tethering Rab8 and Rab11 in the cytosol. PC ablation of TFR1 inhibits parallel fiber-PC LTD, whereas parallel fiber-PC LTP and PC intrinsic excitability are intact. Motor coordination is impaired, but social behaviors are normal in TFR1flox/flox;pCP2-cre mice. Our data reveal a new regulator for trafficking and synaptic expression of mGlu1 and suggest that mGlu1-dependent LTD is associated with motor coordination, but not autistic-like behaviors.


Shine a light: Under-ice light and its ecological implications in a changing Arctic Ocean.
The Arctic marine ecosystem is shaped by the seasonality of the solar cycle, spanning from 24-h light at the sea surface in summer to 24-h darkness in winter. The amount of light available for under-ice ecosystems is the result of different physical and biological processes that affect its path through atmosphere, snow, sea ice and water. In this article, we review the present state of knowledge of the abiotic (clouds, sea ice, snow, suspended matter) and biotic (sea ice algae and phytoplankton) controls on the underwater light field. We focus on how the available light affects the seasonal cycle of primary production (sympagic and pelagic) and discuss the sensitivity of ecosystems to changes in the light field based on model simulations. Lastly, we discuss predicted future changes in under-ice light as a consequence of climate change and their potential ecological implications, with the aim of providing a guide for future research.


Confronting the ethical challenges to informed consent in emergency medicine research.
The Society for Academic Emergency Medicine believes that protection of human subjects is vital in emergency medicine research and that, whenever feasible, informed consent is at the heart of that protection. At the same time, the emergency setting presents unique barriers to informed consent both because of the time frame in which the research is performed and because patients in the emergency department are a vulnerable population. This report reviews the concept of informed consent, empirical data on patients' cognitive abilities during an emergency, the federal rules allowing exemption from consent under certain circumstances, issues surrounding consent forms, and the new Health Insurance Portability and Accountability Act regulations as they relate to research. The authors conclude that, in many circumstances, informed consent is possible if the researcher is diligent and takes time to adequately explain the study to the potential subject. In cases in which it is possible to obtain consent, precautions must be taken to ensure that subjects have decision-making capacity and are offered time to have their questions answered and their needs met. Sometimes resuscitation and other emergency medicine research must be conducted without the ability to obtain consent. In these cases, special protections of subjects under the exception from consent guidelines must be followed. Protection of research subjects is the responsibility of every researcher in emergency medicine.


Monoclonal antibody-based genetic immunotherapy.
The clinical application potential of monoclonal antibodies concerns a wide range of diseases including, among others, viral infections, cancer and autoimmune diseases. Intravenous injection is a simple and obvious mode of administration of purified therapeutic antibodies to patients but may not always be appropriate for long-term treatments for a variety of reasons. One limitation concerns the elevated costs of recombinant proteins certified for human use. Moreover, hour-long infusions require a hospital environment and are often associated with mild to very severe side effects. This makes large-scale clinical applications of a number of monoclonal antibodies with demonstrated therapeutic activity impossible or, at least, severely compromised. In vivo production of therapeutic antibodies in patients, through either genetic modification of their tissues or implantation of antibody-producing cells, might represent an attractive alternative to overcome these drawbacks. Moreover, this method should also provide other benefits. Continuous and sustained delivery of antibodies at a low, but therapeutic level should prevent, or at least delay, induction of neutralizing anti-idiotypic immune responses, which sometimes develop when massive doses of purified immunoglobulins are repeatedly injected into patients. Additionally, it should also limit variations in the bioavailability of therapeutic antibodies that are often detrimental to the efficacy of treatments. The present review reports on the recent developments of gene/cell therapies aiming at the in vivo production and systemic delivery of monoclonal antibodies with the final goal of treating severe chronic diseases.


Betaine concentration as a determinant of fasting total homocysteine concentrations and the effect of folic acid supplementation on betaine concentrations.
Remethylation of homocysteine to methionine can occur through either the folate-dependent methionine synthase pathway or the betaine-dependent betaine-homocysteine methyltransferase pathway. The relevance of betaine as a determinant of fasting total homocysteine (tHcy) is not known, nor is it known how the 2 remethylation pathways are interrelated. The objectives of the study were to examine the relation between plasma betaine concentration and fasting plasma tHcy concentrations and to assess the effect of folic acid supplementation on betaine concentrations in healthy subjects. A double-blind randomized trial of 6 incremental daily doses of folic acid (50-800 microg/d) or placebo was carried out in 308 Dutch men and postmenopausal women (aged 50-75 y). Fasted blood concentrations of tHcy, betaine, choline, dimethylglycine, and folate were measured at baseline and after 12 wk of vitamin supplementation. Concentrations of tHcy were inversely related to the betaine concentration (r = -0.17, P < 0.01), and the association was independent of age, sex, and serum concentrations of folate, creatinine, and cobalamin. Folic acid supplementation increased betaine concentration in a dose-dependent manner (P for trend = 0.018); the maximum increase (15%) was obtained at daily doses of 400-800 microg/d. The plasma betaine concentration is a significant determinant of fasting tHcy concentrations in healthy humans. Folic acid supplementation increases the betaine concentration, which indicates that the 2 remethylation pathways are interrelated.


The Rationale for Exercise in the Management of Pain in Parkinson's Disease.
Pain is a distressing non-motor symptom experienced by up to 85% of people with Parkinson's disease (PD), yet it is often untreated. This pain is likely to be influenced by many factors, including the disease process, PD impairments as well as co-existing musculoskeletal and/or neuropathic pain conditions. Expert opinion recommends that exercise is included as one component of pain management programs; however, the effect of exercise on pain in this population is unclear. This review presents evidence describing the potential influence of exercise on the pain-related pathophysiological processes present in PD. Emerging evidence from both animal and human studies suggests that exercise might contribute to neuroplasticity and neuro-restoration by increasing brain neurotrophic factors, synaptic strength and angiogenesis, as well as stimulating neurogenesis and improving metabolism and the immune response. These changes may be beneficial in improving the central processing of pain. There is also evidence that exercise can activate both the dopaminergic and non-dopaminergic pain inhibitory pathways, suggesting that exercise may help to modulate the experience of pain in PD. Whilst clinical data on the effects of exercise for pain relief in people with PD are scarce, and are urgently needed, preliminary guidelines are presented for exercise prescription for the management of central neuropathic, peripheral neuropathic and musculoskeletal pain in PD.


Why we should use topological data analysis in ageing: Towards defining the "topological shape of ageing".
Living systems are subject to the arrow of time; from birth, they undergo complex transformations (self-organization) in a constant battle for survival, but inevitably ageing and disease trap them to death. Can ageing be understood and eventually reversed? What tools can be employed to further our understanding of ageing? The present article is an invitation for biologists and clinicians to consider key conceptual ideas and computational tools (known to mathematicians and physicists), which potentially may help dissect some of the underlying processes of ageing and disease. Specifically, we first discuss how to classify and analyse complex systems, as well as highlight critical theoretical difficulties that make complex systems hard to study. Subsequently, we introduce Topological Data Analysis - a novel Big Data tool - which may help in the study of complex systems since it extracts knowledge from data in a holistic approach via topological considerations. These conceptual ideas and tools are discussed in a relatively informal way to pave future discussions and collaborations between mathematicians and biologists studying ageing.