Modulation of the TLR4/MyD88/NF-κB Pathway by Humulus japonicus Extract Protects Against Alcohol-Induced Liver Injury in a Rat Model.
Alcohol induces liver injury related to oxidative stress and inflammatory responses. The purpose of this study was to investigate the hepatoprotective effect of Humulus japonicus extract (HJE) against alcohol-induced liver injury. Furthermore, we investigated the mechanisms of the protective effect of HJE on alcohol-induced liver injury. The pretreatment of HJE decreased the levels of aspartate aminotransferase, alanine aminotransferase, triglyceride, and total cholesterol in the plasma, suppressed the malondialdehyde, myeloperoxidase, and enhanced the activities of superoxide dismutase, glutathione, and catalase. The inhibitory effect of HJE against oxidative stress may be associated with the upregulation of nuclear factor erythroid 2-related factor 2 and its target gene heme oxygenase-1. Moreover, HJE inhibited the pro-inflammatory cytokines (tumor necrosis factor alpha, interleukin-1 beta) by downregulating toll-like receptor 4, myeloid differentiation primary response 88, and nuclear factor kappa B p65. These findings provide evidence for the elucidation of the hepatoprotective mechanisms for HJE.


The target tissue in autoimmunity--an influential niche.
Central and peripheral tolerance mechanisms were for a long time the only regulatory circuits known in autoimmunity. It is now becoming clear that the target tissue itself may have the capacity to control its own destiny. Here we review mechanisms by which the target tissue regulates local inflammation, and the way this could influence progression to overt autoimmunity. Moreover, we discuss recent data showing that physiological properties of the target tissue can determine the organ specificity of autoimmune disease. These recent discoveries and ideas concerning the regulatory potential of the target tissue may, in the future, add a new dimension to our concept of regulatory circuits in autoimmunity.


'Oh my god, we're not doing nothing': young people's experiences of spatial regulation.
Social psychologists have become increasingly concerned with examining the ways in which social practices are interrelated with their location. Critical perspectives have highlighted the traditional lack of attention given to both the collective aspects of spatial identities, together with the discursive practices that construct the relationships between people and places. In this article, we draw together the developing discursive work on place with work on children's geographies, in order to examine young people's accounts of spatial regulation. Adopting a discursive approach to the analysis of focus group discussion, we illustrate a variety of concerns managed in relation to spatial practices by 41 young people living in a large city in the South of England. Our findings suggest that everyday use of public space by young people is constructed at a nexus of competing concerns around childhood/adulthood, freedom, and citizenship, and illustrate the dynamic nature of place, and its regulation, as a resource for constructing identities.


Simultaneous traumatic rupture of the patellar tendon and the contralateral quadriceps tendon in a healthy individual.
A simultaneous traumatic complete rupture of the patellar tendon and the contralateral quadriceps tendon is reported to occur in patients with renal failure and other inflammatory diseases, but is extremely rare in a healthy individual because of the different contributory factors and mechanisms of injury. We present a rare case report of such a combination of injuries in a 48-year-old healthy man. To our knowledge only three such cases have been reported in the English literature. This is an unusual combination and hence there is potential for missed diagnosis leading to suboptimal treatment.


Monoclonal antibodies against rat leukocyte surface antigens.
Monoclonal antibodies have proven to be powerful tools for studying the properties of leukocyte surface antigens and the cells that express them. In the past decades many monoclonal antibodies (mAb) for identifying the different rat leukocyte surface antigens have been described. A list of mAb is provided in Table I below. The rat leukocyte surface antigens are divided into different sections, including rat CD antigens (a), rat leukocyte surface antigens without CD designation (b), rat major histocompatibility complex (MHC) antigens (c), rat T-cell receptors (d) and rat immunoglobulins (e). The molecular and functional characteristics of rat leukocyte surface antigens are discussed in more detail in some of the other chapters of this issue (e.g. Van den Berg et al., p. 45). A more extensive overview of the properties of leukocyte surface antigens is provided by Barclay et al.


Effect of Age on the Initiation of Biologic Agent Therapy in Patients With Inflammatory Bowel Disease: Korean Common Data Model Cohort Study.
The Observational Health Data Sciences and Informatics (OHDSI) network is an international collaboration established to apply open-source data analytics to a large network of health databases, including the Korean common data model (K-CDM) network. The aim of this study is to analyze the effect that age at diagnosis has on the prognosis of inflammatory bowel disease (IBD) in Korea using a CDM network database. We retrospectively analyzed the K-CDM network database from 2005 to 2015. We transformed the electronic medical record into the CDM version 5.0 used in OHDSI. A worsened IBD prognosis was defined as the initiation of therapy with biologic agents, including infliximab and adalimumab. To evaluate the effect that age at diagnosis had on the prognosis of IBD, we divided the patients into an early-onset (EO) IBD group (age at diagnosis <40 years) and a late-onset (LO) IBD group (age at diagnosis ≥40 years) with the cutoff value of age at diagnosis as 40 years, which was calculated using the Youden index method. We then used the logrank test and Cox proportional hazards model to analyze the effect that age at diagnosis (EO group vs LO group) had on the prognosis in patients with IBD. A total of 3480 patients were enrolled. There was 2017 patients with ulcerative colitis (UC) and 1463 with Crohn's disease (CD). The median follow up period was 109.5 weeks. The EO UC group was statistically significant and showed less event-free survival (ie, experiences of biologic agents) than the LO UC group (P<.001). In CD, the EO CD group showed less event-free survival (ie, experiences of biologic agents) than the LO CD group. In the Cox proportional hazard analysis, the odds ratio (OR) of the EO UC group on experiences of biologic agents compared with the LO UC group was 2.3 (95% CI 1.3-3.8, P=.002). The OR of the EO CD group on experiences of biologic agents compared with the LO CD group was 5.4 (95% CI 1.9-14.9, P=.001). The EO IBD group showed a worse prognosis than the LO IBD group in Korean patients with IBD. In addition, this study successfully verified the CDM model in gastrointestinal research.


Random Search with Resetting: A Unified Renewal Approach.
We provide a unified renewal approach to the problem of random search for several targets under resetting. This framework does not rely on specific properties of the search process and resetting procedure, allows for simpler derivation of known results, and leads to new ones. Concentrating on minimizing the mean hitting time, we show that resetting at a constant pace is the best possible option if resetting helps at all, and derive the equation for the optimal resetting pace. No resetting may be a better strategy if without resetting the probability of not finding a target decays with time to zero exponentially or faster. We also calculate splitting probabilities between the targets, and define the limits in which these can be manipulated by changing the resetting procedure. We moreover show that the number of moments of the hitting time distribution under resetting is not less than the sum of the numbers of moments of the resetting time distribution and the hitting time distribution without resetting.


Somatostatin and Thyrotrophin-Releasing Hormone Response and Receptor Status of a Thyrotrophin-Secreting Pituitary Adenoma: Clinical and in vitro Studies.
Abstract We have described a patient with a thyrotrophin-secreting pituitary adenoma and correlated a detailed physiological and anatomical investigation of the surgically resected tumour with its in vivo regulation. Thyrotrophin secretion was inhibited by circulating thyroid hormones, dopaminergic agonists and the somatostatin analogue SMS 201-995 but could not be stimulated by thyrotrophin-releasing hormone or further inhibited by exogenous triiodothyronine. Prolonged treatment with SMS 201-995 caused tumour shrinkage as shown by successive computed tomography scans but was accompanied by tumour desensitization and the development of diabetes mellitus. This is the first thyrotroph adenoma in which somatostatin receptors have been directly demonstrated and shown to completely block thyrotrophin-releasing hormone-induced inositol phospholipid accumulation when occupied. In addition, preincubation with triiodothyronine significantly inhibited thyrotrophin-releasing hormone-induced inositol phospholipid turnover in dispersed pituitary cells indicating that in this tumour, circulating thyroid hormones were exerting feedback inhibition at the level of the pituitary either by reducing the number of thyrotrophin-releasing hormone receptors and/or their coupling to second messenger pathways. In keeping with this hypothesis, the acute reduction in intrapituitary triiodothyronine by sodium ipodate in vivo had no effect on peripheral thyrotrophin over 6 h suggesting that the onset of the effect of triiodothyronine withdrawal on thyrotrophin secretion was suitably delayed. The importance of the inositol phospholipid second messenger pathway in transducing the secretory response in this tumour was further corroborated by electrophysiological studies which demonstrated thyrotrophin-releasing hormone-induced changes in K(+) currents which are dependent on intracellular Ca(2+) ions, presumably mobilized via the inositol phospholipids. In addition to thyrotrophin and alpha subunit, growth hormone mRNA and growth hormone were found throughout the tumour as were two populations of cells distinguished electron microscopically by the size of their secretory granules. Although acromegalic features are not unusual in thyrotroph adenomas, our patient showed no evidence of inappropriate growth hormone secretion during surgery or in response to pre- or post-operative insulin stress tests.


Menopausal age, postmenopausal hormone therapy and incident atrial fibrillation.
Limited data exist on the association between menopause and atrial fibrillation (AF). We sought to examine the relationship between menopausal age, postmenopausal hormone therapy (PHT) use and incident AF. The Women's Health Study (WHS) enrolled 39 876 female health professionals between 1992 and 1995. We prospectively examined 30 034 women in WHS using Cox proportional-hazard models. Participants were free of cardiovascular disease and AF at baseline and had not undergone hysterectomy without bilateral oophorectomy prior to menopause. Incident AF was confirmed by medical record review. At baseline, median age was 53 years (IQR 49-60), median menopausal age was 50 years (IQR 46-52) and 14 415 (48.0%) had prior PHT use. Over a median follow-up of 20.5 years, 1350 AF events occurred. In multivariable analysis, relative hazards for AF were lower among women with younger age at menopause but did not differ significantly from women with the oldest menopausal age (<45: HR 0.82, 95% CI 0.67 to 1.02; 45-49: HR 0.90, 95% CI 0.74 to 1.08; 50-54: HR 0.89, 95% CI 0.75 to 1.06; >54 years: referent). Use of oestrogen-alone PHT, but not oestrogen and progesterone, was independently associated with AF risk (HR 1.22; 95% CI 1.02 to 1.45 vs HR 1.04; 95% CI 0.86 to 1.26). This relationship was not attenuated by intermediary cardiovascular events. In this large prospective study, menopausal age was not significantly related to incident AF, while use of oestrogen monotherapy was associated with increased AF risk. Our findings suggest a pathophysiological link between unopposed oestrogen exposure and AF in women. NCT000000479; Post-results.


Microbiota-Gut-Brain Axis: Modulator of Host Metabolism and Appetite.
The gut harbors an enormous diversity of microbes that are essential for the maintenance of homeostasis in health and disease. A growing body of evidence supports the role of this microbiota in influencing host appetite and food intake. Individual species within the gut microbiota are under selective pressure arising from nutrients available and other bacterial species present. Each bacterial species within the gut aims to increase its own fitness, habitat, and survival via specific fermentation of dietary nutrients and secretion of metabolites, many of which can influence host appetite and eating behavior by directly affecting nutrient sensing and appetite and satiety-regulating systems. These include microbiota-produced neuroactives and short-chain fatty acids. In addition, the gut microbiota is able to manipulate intestinal barrier function, interact with bile acid metabolism, modulate the immune system, and influence host antigen production, thus indirectly affecting eating behavior. A growing body of evidence indicates that there is a crucial role for the microbiota in regulating different aspects of eating-related behavior, as well as behavioral comorbidities of eating and metabolic disorders. The importance of intestinal microbiota composition has now been shown in obesity, anorexia nervosa, and forms of severe acute malnutrition. Understanding the mechanisms in which the gut microbiota can influence host appetite and metabolism will provide a better understanding of conditions wherein appetite is dysregulated, such as obesity and other metabolic or eating disorders, leading to novel biotherapeutic strategies.


Informatics in radiology: use of a macro scripting editor to facilitate transfer of dual-energy X-ray absorptiometry reports into an existing departmental voice recognition dictation system.
The process of verbally reporting or manually retyping numeric data generated at dual-energy x-ray absorptiometry (DXA) involves numerous pitfalls. With use of a macro scripting editor, a customized macro was created to automate the transfer of data generated by a DXA scanner into a structured voice recognition dictation system without requiring radiologists to type in a medical record number or accession number to identify the study. A preliminary report is generated with use of software for a DXA unit and a customized template that includes numeric and qualitative assessments of osteoporosis as well as data from prior studies if available. A customized macro is then invoked by the macro scripting editor, which selectively transfers the report from the draft document into the voice recognition dictation system, thereby producing a final structured diagnostic report. All of the radiologists surveyed to evaluate this automated method reported ease of software use and greater efficiency in report production. In addition, a random audit of the 800 DXA scans that have been reported with this technique demonstrated no reports generated under an incorrect accession number and no incorrect transfer of data. Automated DXA reporting is now the preferred method of dictation at the authors' institution and represents an inexpensive, accurate, and customizable means of DXA reporting.


Osteogenic protein-1, a bone morphogenetic protein, induces angiogenesis in the chick chorioallantoic membrane and synergizes with basic fibroblast growth factor and transforming growth factor-beta1.
Capillary invasion is a vital regulatory signal during bone morphogenesis that is influenced by angiogenic molecules such as fibroblast growth factor (FGF) and some members of the transforming growth factor-beta (TGF-beta) superfamily, including TGF-betas themselves. Bone morphogenetic proteins (BMPs), which are members of the TGF-beta superfamily, have previously not been shown to possess direct angiogenic properties. Osteogenic protein-1 (OP-1; BMP-7) is a potent regulator of cartilage and bone differentiation in vivo. The osteogenic and angiogenic properties of OP-1 at both ortho- and heterotopic sites in adult chacma baboons (Papio ursinus) are enhanced synergistically by the simultaneous application of relatively low doses of TGF-beta1. The single application of relatively high doses of TGF-beta1 (20 ng), and bFGF (500 ng) or relatively low (100 ng) and high (1,000 ng) doses of OP-1 in the chick chorioallantoic membrane (CAM) assay elicited a prominent and (for OP-1) dose-dependent angiogenic response. The binary application of a relatively low dose of OP-1 (100 ng) with a relatively low dose of bFGF (100 ng) or with a relatively low (5 ng) or high (20 ng) dose of TGF-beta1 resulted in a synergistic enhancement of the angiogenic response. The angiogenic effect of the relatively low doses of the combined morphogens was distinctly more pronounced than that of the single application of the relatively high doses of the respective factors. The present findings suggest that these morphogens may be deployed in binary combination in order to accentuate experimental angiogenesis. The cooperative interaction of the different morphogens in the CAM assay may provide important biological clues towards the control of clinical angiogenesis.


Building triangular nanoprisms from the bottom-up: a polyelectrolyte micellar approach.
Charge-regulated synthesis of triangular prisms in aqueous solution using self-assembled polyelectrolyte micelles as templates is described in detail. Micelles formed from amphiphilic polystyrene-block-sulfonated poly(1,3-cyclohexadiene) (PS-b-sPCHD) serve as templates to direct the formation of novel triangular prisms of CuCl2 single crystals. We demonstrate that the edge lengths of these triangular prisms can be easily tailored at room temperature from the nanoscale to the mesoscale by simply adjusting the ratio of charged micelles to protons in the solution. This approach can be extended to the preparation of different ordered crystal structures with a precision hard to achieve via other approaches.


Porokeratosis of Mibelli. A perforating disease of the epidermis.
A case of linear porokeratosis of Mibelli is presented. Although there was no suggestion clinically of a perforating disorder, histologic examination revealed a cornoid lamella penetrating into the dermis. Special stains did not show any evidence of a transepidermal elimination syndrome.


A study of drug abuse in a group of South African university students.
Seventeen informal interviews and 1597 questionnarie responses provided an investigation of drug abuse in a group of South African students. Legal problems in South Africa make research of drug abuse (and indeed many other areas of social relations), particularly interviewing, extremely hazardous and therefore uncommon. Not surprisingly, in view of this, the interviews revealed findings inconsistent with the questionnaire data in many areas. Indeed, the informal technique demonstrated the need for more personal research in this field rather than grouped data questionnaires, the latter being seemingly responsible for the myth of the stereotypic drug taker.


Identification of Ser153 in ICL2 of the gonadotropin-releasing hormone (GnRH) receptor as a phosphorylation-independent site for inhibition of Gq coupling.
Type I gonadotropin-releasing hormone (GnRH) receptor (GnRHR) is unique among mammalian G-protein-coupled receptors (GPCRs) in lacking a C-terminal tail, which is involved in desensitization in GPCRs. Therefore, we searched for inhibitory sites in the intracellular loops (ICLs) of the GnRHR. Synthetic peptides corresponding to the three ICLs were inserted into permeabilized alphaT3-1 gonadotrope cells, and GnRH-induced inositol phosphate (InsP) formation was determined. GnRH-induced InsP production was potentiated by ICL2 > ICL3 but not by the ICL1 peptides, suggesting they are acting as decoy peptides. We examined the effects of six peptides in which only one of the Ser or Thr residues was substituted with Ala or Glu. Only substitution of Ser153 with Ala or Glu ablated the potentiating effect upon GnRH-induced InsP elevation. ERK activation was enhanced, and the rate of GnRH-induced InsP formation was about 6.5-fold higher in the first 10 min in COS-1 cells that were transfected with mutants of the GnRHR in which the ICL2 Ser/Thr residues (Ser151, Ser153, and Thr142) or only Ser153 was mutated to Ala as compared with the wild type GnRHR. The data indicate that ICL2 harbors an inhibitory domain, such that exogenous ICL2 peptide serves as a decoy for the inhibitory site (Ser153) of the GnRHR, thus enabling further activation. GnRH does not induce receptor phosphorylation in alphaT3-1 cells. Because the phosphomimetic ICL2-S153E peptide did not mimic the stimulatory effect of the ICL2 peptide, the inhibitory effect of Ser153 operates through a phosphorylation-independent mechanism.


Epidermal growth factor down-regulates the expression of human hepatic stimulator substance via CCAAT/enhancer-binding protein β in HepG2 cells.
hHSS (human hepatic stimulator substance), acting as a hepatotrophic growth factor, promotes liver regeneration. However, the regulatory mechanisms for hHSS transcription are still poorly understood. In the present study, we investigated transcription of hHSS triggered by EGF (epidermal growth factor) and the role of C/EBPβ (CCAAT/enhancer-binding protein β) as a potential core factor responsible for hHSS transcription in HepG2 cells. The results show that EGF suppresses hHSS mRNA expression at early time points. Using a promoter deletion assay, we identified a proximal region (-358/-212) that is required for EGF suppression. Overexpression of C/EBPβ enhances EGF suppression of hHSS, and mutation of the C/EBPβ-binding site at -292/-279 or siRNA (short interfering RNA) interference abolishes EGF suppression. Furthermore, using an electrophoretic mobility-shift assay and chromatin immunoprecipitation analysis, we found that C/EBPβ specifically binds to the -292/-279 site that is responsible for EGF inhibition. Moreover, using a knockin (overexpression) and knockdown strategy (siRNA), we confirmed that C/EBPβ is a key factor responsible for inhibition of hHSS mRNA expression. Pre-treatment with an inhibitor of JNK (c-Jun N-terminal kinase) or down-regulation of JNK1 with specific siRNA reverses EGF-inhibited hHSS expression. Our results provide a crucial regulatory mechanism for EGF in hHSS transcription within the promoter proximal region.


Burn injury results in decreased gastric acid production in the acute shock period.
Gastrointestinal complications including acute stress ulcerations occur after burn injury. The causes of acute gastric derangements are multiple, and tissue ischemia in the acute period of burn shock may promote breakdown of the gastric mucosal protective barrier. We compared gastric pH in mice after 25% total body surface area, full-thickness murine burn injury with that in unburned control animals. Animals were anesthetized with methoxyflurane and were resuscitated with 1 ml normal saline solution immediately after burn. Animals were killed at intervals up to 24 hours after burn injury, stomachs were removed and opened, and gastric mucosal pH was measured by use of a surface pH probe. In other animals mixed venous blood was obtained via direct inferior vena cava puncture, and blood gas analysis was performed at intervals up to 24 hours after burn injury. Unexpectedly, gastric mucosal pH increased in burned mice compared with that in controls. The peak difference, greater than one log pH unit, occurred at 3 hours after burn injury (pH 4.45 burn vs pH 3.34 control, p < 0.00001), and this difference was maintained through 12 hours (pH 4.88 burn vs pH 3.20 control, p < 0.005). In this model, shock was observed to begin as early as 1 hour after burn injury and reached its maximal period (base deficit, -27.8 mEq/L) at 12 hours after burn injury. In view of the higher gastric pH in burned mice with concomitant profound shock, gastric acid production appears to be impaired during this time, which suggests acute postburn gastrointestinal ulcerations may be primarily due to ischemia. Prevention of organ ischemia may play a key role in the prevention of acute gastric ulcerations after burn injury.


Transcriptional profiles underlying parent-of-origin effects in seeds of Arabidopsis thaliana.
Crossing plants of the same species but different ploidies can have dramatic effects on seed growth, but little is known about the alterations to transcriptional programmes responsible for this. Parental genomic imbalance particularly affects proliferation of the endosperm, with an increased ratio of paternally to maternally contributed genomes ('paternal excess') associated with overproliferation, while maternal excess inhibits endosperm growth. One interpretation is that interploidy crosses disrupt the balance in the seed of active copies of parentally imprinted genes. This is supported by the observation that mutations in imprinted FIS-class genes of Arabidopsis thaliana share many features of the paternal excess phenotype. Here we investigated gene expression underlying parent-of-origin effects in Arabidopsis through transcriptional profiling of siliques generated by interploidy crosses and FIS-class mutants. We found that fertilized fis1 mutant seeds have similar profiles to seeds with paternal excess, showing that the shared phenotypes are underpinned by similar patterns of gene expression. We identified genes strongly associated with enhanced or inhibited seed growth; this provided many candidates for further investigation including MADS-box transcription factors, cell cycle genes, and genes involved in hormone pathways. The work presented here is a step towards understanding the effects on seed development of the related phenomena of parental genome balance and imprinting.


RUNX-1 haploinsufficiency causes a marked deficiency of megakaryocyte-biased hematopoietic progenitor cells.
Patients with familial platelet disorder with a predisposition to myeloid malignancy (FPDMM) harbor germline monoallelic mutations in a key hematopoietic transcription factor, RUNX-1. Previous studies of FPDMM have focused on megakaryocyte (Mk) differentiation and platelet production and signaling. However, the effects of RUNX-1 haploinsufficiency on hematopoietic progenitor cells (HPCs) and subsequent megakaryopoiesis remains incomplete. We studied induced pluripotent stem cell (iPSC)-derived HPCs (iHPCs) and Mks (iMks) from both patient-derived lines and a wild-type (WT) line modified to be RUNX-1 haploinsufficient (RUNX-1+/-), each compared with their isogenic WT control. All RUNX-1+/- lines showed decreased iMk yield and depletion of an Mk-biased iHPC subpopulation. To investigate global and local gene expression changes underlying this iHPC shift, single-cell RNA sequencing was performed on sorted FPDMM and control iHPCs. We defined several cell subpopulations in the Mk-biased iHPCs. Analyses of gene sets upregulated in FPDMM iHPCs indicated enrichment for response to stress, regulation of signal transduction, and immune signaling-related gene sets. Immunoblot analyses in FPDMM iMks were consistent with these findings, but also identified augmented baseline c-Jun N-terminal kinase (JNK) phosphorylation, known to be activated by transforming growth factor-β1 (TGF-β1) and cellular stressors. These findings were confirmed in adult human CD34+-derived stem and progenitor cells (HSPCs) transduced with lentiviral RUNX1 short hairpin RNA to mimic RUNX-1+/-. In both iHPCs and CD34+-derived HSPCs, targeted inhibitors of JNK and TGF-β1 pathways corrected the megakaryopoietic defect. We propose that such intervention may correct the thrombocytopenia in patients with FPDMM.


[Neurocysticercosis. Diagnostic and therapeutic advances].
A 42-year-old man of Croatian birth, with long-standing neurological symptoms, was found to have neurocysticercosis (a rare disease in Central Europe), as proven by positive antibody titres in serum and CSF and typical foci by computed tomography (CT). Ventriculo-cisternal drainage (after Torkildsen) controlled the acute symptoms of raised intracranial pressure. Postoperatively the CT revealed new low-density intracerebral foci, which responded to praziquantel.


Moment-angle relations after specific exercise.
This study examined the amount and time-course of shifts in the moment-knee angle relation of the quadriceps (QF) and hamstring (HAM) muscles in response to different length-restricted strength training regimens. Thirty-two athletes were divided into three different training groups (G1-3): G1 performed isometric training at knee joint angles corresponding to long muscle-tendon unit (MTU) length for QF and HAM; G2 conducted concentric-eccentric contraction cycles that were restricted to a knee joint range of motion corresponding to predominantly long MTU length for QF and HAM; G3 combined the protocols of G1 and G2. Moment-knee angle and EMG-knee angle relations of QF and HAM were measured on five different occasions: two times before, after five and eight weeks of training and four weeks post training. Moments and EMG-data of each subject were normalized to the largest value produced at any knee joint position [% Max.]. Obtained by curve fitting, the optimal knee joint angle for QF moment production was significantly (P<0.05) shifted to longer MTU length in G1 and G3 after 5 weeks of training and in G2 after 8 weeks of training. Contrary, no significant shifts were detected for HAM. Our data suggest that the predominant MTU length during loading is a major trigger for human force-length adaptations.


N-Glycan processing by a lepidopteran insect alpha1,2-mannosidase.
Protein glycosylation pathways are relatively poorly characterized in insect cells. As part of an overall effort to address this problem, we previously isolated a cDNA from Sf9 cells that encodes an insect alpha1,2-mannosidase (SfManI) which requires calcium and is inhibited by 1-deoxymannojirimycin. In the present study, we have characterized the substrate specificity of SfManI. A recombinant baculovirus was used to express a GST-tagged secreted form of SfManI which was purified from the medium using an immobilized glutathione column. The purified SfManI was then incubated with oligosaccharide substrates and the resulting products were analyzed by HPLC. These analyses showed that SfManI rapidly converts Man(9)GlcNAc(2)to Man(6)Glc-NAc(2)isomer C, then more slowly converts Man(6)GlcNAc(2)isomer C to Man(5)GlcNAc(2). The slow step in the processing of Man(9)GlcNAc(2)to Man(5)GlcNAc(2)by SfManI is removal of the alpha1,2-linked mannose on the middle arm of Man(9)GlcNAc(2). In this respect, SfManI is similar to mammalian alpha1,2-mannosidases IA and IB. However, additional HPLC and(1)H-NMR analyses demonstrated that SfManI converts Man(9)GlcNAc(2)to Man(5)GlcNAc(2)primarily through Man(7)GlcNAc(2)isomer C, the archetypal Man(9)GlcNAc(2)missing the lower arm alpha1,2-linked mannose residues. In this respect, SfManI differs from mammalian alpha1,2-mannosidases IA and IB, and is the first alpha1,2-mannosidase directly shown to produce Man(7)GlcNAc(2)isomer C as a major processing intermediate.


Sexual dimorphism in adult Little Stints (Calidris minuta) revealed by DNA sexing and discriminant analysis.
The sex of an individual organism plays such an important role in its life cycle that researchers must know a bird's sex to interpret key aspects of its biology. The sexes of dimorphic species can be easily distinguished, but sexing monomorphic bird species often requires expensive and time-consuming molecular methods. The Little Stint (Calidris minuta) is a numerous species, monomorphic in plumage but showing a small degree of reversed sexual size dimorphism. Females are larger than males but the ranges of their measurements overlap, making Little Stints difficult to sex in the field. Our aim was to develop reliable sexing criteria for Little Stints in different stages of primary moult during their stay on the non-breeding grounds in South Africa using DNA-sexed individuals and discriminant function analysis. We caught 348 adult Little Stints in 2008-2016 on their non-breeding grounds at Barberspan Bird Sanctuary. To molecularly identify the birds' sex we used P2/P8 primers and DNA isolated from blood samples collected in the field. We used Storer's dimorphism index to assess the degree of sexual size dimorphism. Then we divided our sample into two groups: before or during and after primary moult. For each group we developed two functions: one using wing length only and the other a combination of morphometric features including wing, tarsus and total head length. Then we used a stepwise procedure to check which combination of measurements best discriminated sexes. To validate our result we used a jack-knife cross-validation procedure and Cohen-kappa statistics. All the morphometric features we measured were bigger in DNA-sexed females than in males. Birds with fresh primaries had on average 2.3 mm longer wings than those with worn primaries. A discriminant function using wing length (D1) correctly sexed 78.8% of individuals before moult, and a stepwise analysis showed that a combination of wing length and tarsus (D2) correctly identified the sex of 82.7% of these birds. For birds with freshly moulted primaries a function using wing length (D3) correctly classified 83.4% of the individuals, and a stepwise analysis revealed that wing and total head length (D4) classified 84.7%. Sexual size differences in Little Stints might be linked to their phylogenetics and breeding biology. Females are bigger, which increases their fecundity; males are smaller, which increases their manoeuverability during display flights and hence their mating success. Little Stints show an extreme lack of breeding site fidelity so we did not expect a geographical cline in their biometrics. Sexing criteria available for Little Stints in the literature were developed using museum specimens, which often shrink, leading to misclassification of live birds. The sexing criteria we developed can be used for studies on Little Stints at their non-breeding grounds and on past data, but should be applied cautiously because of the overlapping ranges.


Facile and rapid ruthenium mediated photo-crosslinking of Bombyx mori silk fibroin.
A facile and rapid ruthenium catalysed photo-crosslinking method was employed to prepare Bombyx mori silk fibroin hydrogels for the first time that may be used for biomedical applications. The gels have been characterised by differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), photo-acoustic Fourier transform infrared spectroscopy (PA-FTIR), and X-ray diffraction (XRD). We have reported the tuning of the properties of the chemically crosslinked gels through hydration level. DSC and DMA measurements have shown that the molecular chain dynamics and mechanical properties are dependent on the moisture/water content. The structural changes upon gelation were examined through the use of DSC, PA-FTIR and XRD, which confirmed that the gels were less crystalline than the original silk powder. Finally, the biocompatibility and hence, the potential tissue engineering application of the silk fibroin hydrogel was also assessed by culturing chondrocyte progenitor cells for up to one week on the engineered fibroin surfaces.


Eating with others and meal location are differentially associated with nutrient intake by sex: The Diabetes Study of Northern California (DISTANCE).
Though eating with others is often a social behavior, relationships between social contexts of eating and nutrient intake have been underexplored. This study evaluates how social aspects of eating - frequencies of eating meals with others, meals prepared at home, and meals outside the home - are associated with nutrient intake. Because diet improvement can reduce complications of diabetes mellitus, we surveyed a multi-ethnic cohort of persons with type 2 diabetes (n = 770) about social aspects of diet (based on 24-hour recalls). Sex-stratified multiple regression analyses adjusted for confounders assessed the relationship between frequency of eating with others and nutrient intake (total energy, energy from fat, energy from carbohydrates, Healthy Eating Index/HEI, Dietary Approaches to Stop Hypertension/DASH score). Although there was slight variation in men's versus women's propensity to share meals, after adjustment for confounders, there was no consistently significant association between meals with others and the 5 nutrient intake measures for either men or women. The directions of association between categories of eating with others and diet quality (HEI and DASH scores) - albeit not significant - were different for men (positive) and women (mostly negative), which warrants further investigation. The next analyses estimated nutrient intake associated with meals prepared at home, and meals consumed outside the home. Analyses indicated that greater meal frequency at home was associated with significantly better scores on diet quality indices for men (but not women), while meal frequency outside the home was associated with poorer diet quality and energy intake for women (but not men). Better measurement of social dimensions of eating may inform ways to improve nutrition, especially for persons with diabetes for whom diet improvement can result in better disease outcomes.


Peripheral cannulation for cardiopulmonary bypass.
Peripheral cannulation is the historical route for connecting the pump-oxygenator to the vasculature of the patient in order to establish partial or complete cardiopulmonary bypass. Although most open heart procedures are nowadays realized with central cannulation, there is renewed interest in remote cannulation through the femoral, iliac, axillary, subclavian and jugular vessels. Remote cannulation is not only of interest in hemodynamically unstable patients who can be put on cardiopulmonary bypass in local anesthesia, and stabilized prior to intubation, but also for complex procedures like replacement of the thoracoabdominal aorta, acute type A aortic dissections, complex redo open heart surgery, extracorporeal membrane oxygenation, and more recently, small access open heart surgery, robotic surgery, and others. In the following shall be described femoral arterial cannulation with standard cannulas, femoral arterial cannulation with percutaneous cannulas, femoral venous cannulation using standard cannulas, femoral venous cannulation using percutaneous cannulas, as well as optimized venous cannulation relying on more advanced cannula designs.


Anti-Inflammatory and Chondroprotective Effects Induced by Phenolic Compounds from Onion Waste Extracts in ATDC-5 Chondrogenic Cell Line.
Osteoarthritis is a prevalent degenerative condition that is closely related to the destruction and inflammation of cartilage. The high prevalence of this pathology exhorts researchers to search for novel therapeutic approaches. Vegetable-fruit wastes have emerged as a promising origin of anti-inflammatory and antioxidant compounds that, in some cases, may also exert chondroprotective effects. This study aims to decipher the potential of onion waste products in the inhibition of molecular events involved in osteoarthritis. Onion extracts showed a high content of phenolic compounds and antioxidant properties. Cytocompatibility was demonstrated in the chondrogenic cell line ATDC-5, exerting viability percentages higher than 90% and a slight increase in the S phase cycle cell. The induction of inflammation mediated by the lipopolysaccharide and onion extracts' treatment substantially inhibited molecular markers related to inflammation and cartilage degradation, highlighting the promising application of onion extracts in biomedical approaches. The in silico analyses suggested that the results could be attributed to protocatechuic, ellagic, and vanillic acids' greater cell membrane permeability. Our work provides distinctive information about the possible application of waste onion extracts as functional components with anti-inflammatory and chondroprotective characteristics in osteoarthritis.


Towards a characterisation of the wild legume bitter vetch (Lathyrus linifolius L. (Reichard) Bässler): heteromorphic seed germination, root nodule structure and N-fixing rhizobial symbionts.
Lathyrus linifolius L. (Reichard) Bässler (Fabiaceae, bitter vetch) is a nitrogen (N) fixing species. A coloniser of low nutrient (N) soils, it supports biodiversity such as key moth and butterfly species, and its roots are known for their organoleptic and claimed therapeutic properties. Thus, the species has high potential for restoration, conservation, novel cropping and as a model species. The last because of its genetic synteny with important pulse crops. However, regeneration and functional attributes of L. linifolius remain to be characterised. Seeds of L. linifolius were characterised using physical, colorimetric and chemical data. Ultrastructural and functional characterisation of the N-fixing root nodules included immunolabelling with nifH protein antibodies (recognising the N-fixing enzyme, nitrogenase). Endosymbiotic bacteria were isolated from root nodules and characterised phylogenetically using 16S rRNA, nodA and nodD gene sequences. L. linifolius yielded heteromorphic seed of distinct colour classes: green and brown. Seed morphotypes had similar C:N ratios and were equally germinable (ca. 90%) after scarification at differing optimal temperatures (16 and 20 °C). Brown seeds were larger and comprised a larger proportion of the seed batch (69%). L. linifolius root nodules appeared indeterminate in structure, effective (capable of fixing atmospheric N) and having strains very similar to Rhizobium leguminosarum biovar viciae. The findings and rhizobial isolates have potential application for ecological restoration and horticulture using native seeds. Also, the data and rhizobial resources have potential applications in comparative and functional studies with related and socio-economically important crops such as Pisum, Lens and Vicia.


Enhancing GABAergic Transmission Improves Locomotion in a Caenorhabditis elegans Model of Spinal Muscular Atrophy.
Spinal muscular atrophy (SMA) is a neuromuscular disease characterized by degeneration of spinal motor neurons resulting in variable degrees of muscular wasting and weakness. It is caused by a loss-of-function mutation in the survival motor neuron (SMN1) gene. Caenorhabditis elegans mutants lacking SMN recapitulate several aspects of the disease including impaired movement and shorted life span. We examined whether genes previously implicated in life span extension conferred benefits to C. elegans lacking SMN. We find that reducing daf-2/insulin receptor signaling activity promotes survival and improves locomotor behavior in this C. elegans model of SMA. The locomotor dysfunction in C. elegans lacking SMN correlated with structural and functional abnormalities in GABAergic neuromuscular junctions (NMJs). Moreover, we demonstrated that reduction in daf-2 signaling reversed these abnormalities. Remarkably, enhancing GABAergic neurotransmission alone was able to correct the locomotor dysfunction. Our work indicated that an imbalance of excitatory/inhibitory activity within motor circuits and underlies motor system dysfunction in this SMA model. Interventions aimed at restoring the balance of excitatory/inhibitory activity in motor circuits could be of benefit to individuals with SMA.


Sedation, analgesia and anesthesia for interventional radiological procedures in adults. Part II. Recommendations for interventional radiologists.
Benzodiazepines are given orally as a premedication before an interventional radiological procedure. Local analgesia is achieved by drugs such as lidocaine, bupivacaine or ropivacaine. General analgesia is obtained by non opioid analgesics and opioid narcotics. For intravenous sedation, benzodiazepines such as ketamine or propofol should be administered under the supervision of an anesthesiologist. A preprocedure consultation with the anesthesiologist is recommended. Monitoring equipments, drugs and nursing staff assistance should be provided in the interventional suite. Vital signs should be monitored for several hours until patient's discharge. Close collaboration between anesthesiologists and interventional radiologists is a prerequisite for achieving high standard sedation and analgesia.


Planum temporale analysis via a new volumetric method in autoptic brains of demented and psychotic patients.
Investigations of alterations in brain asymmetry often focus on the planum temporale of patients with schizophrenia. Data also suggest changes in laterality of demented patients associated with a more marked impairment of the left hemisphere. Our study was performed on autoptic brain tissue of 84 patients, out of which there were 25 non-demented non-psychotic controls, 50 demented patients (34 Alzheimer disease, 9 multi - infarct dementia and 7 mixed-type dementia patients) and 9 people with schizophrenia. The plana temporalia were evaluated via a new volumetric method using dental resin matter. Areas, cortical thickness and volumes of the right and left planum temporale were evaluated without normalization to brain weight in 60 patients and with normalization in 24 people. In controls, a mild right/left laterality of areas, cortical thickness and volumes was found. Moreover, in control women the areas of the left planum temporale were smaller than those observed in control men. The shifts to left/right laterality of areas and volumes were found in all demented groups. In the more numerous Alzheimer group, the change in laterality of an area was associated with a mild decrease on the right and a mild increase on the left side. In contrast, marked but only bilateral area shrinkage as well as reduced cortical thickness and brain volumes were observed in schizophrenic patients.


The Effects of Gender on Radiation Dose during Fenestrated Endovascular Aneurysm Repair.
Female gender is considered a risk factor for worse perioperative outcomes after fenestrated endovascular aneurysm repair (FEVAR). We hypothesized that women would have more unfavorable anatomy, increasing case complexity and leading to higher radiation doses. Our aim was to evaluate the effect of gender on radiation dose during FEVARs. This single-center retrospective study was performed from 1/2015 to 2/2018. For patient data, linear model and stepwise variable selection algorithm were used. All dose measurements were log transformed before analysis. Significance level for parameter estimates and corresponding 95% confidence intervals were all transformed back using an exponential function. P-value of <0.05 was considered statistically significant. All analyses were performed in SAS 9.4 (SAS Institute Inc., Cary, NC). A total of 169 FEVARs (45 women) were performed on a Philips Allura Xper FD 20 fluoroscopy system equipped with clarity technology. There was no difference in body mass index (BMI) or operative time between genders, P = 0.9. The median reference air kerma for women was significantly lower than that for men (1,672 mGy vs. 2,496 mGy), P < 0.001. Women had on average a 28% total dose reduction after controlling for BMI, number of vessels fenestrated, operative time, and type of device, P < 0.001. The median fluorography and fluoroscopy doses for women were significantly lower than those for men (973 mGy vs. 1,401 mGy and 659 mGy vs. 1,008 mGy), resulting in a 24% fluorography dose reduction and a 38% fluoroscopy dose reduction for women, P < 0.001. FEVARs can be performed successfully in women with comparatively lower radiation doses.


[Health literacy: translation and validation of a research instrument on health promotion in Brazil].
The aim of this study was to translate, culturally adapt, and test the psychometric performance of a questionnaire to assess health literacy. Brazilian university students (n = 472) with a mean age of 22.7 (5.3) years participated in the study. The validities of the factor, convergent, and discriminant structure were tested using structural equations analysis. The 4-factors model showed only fair results, but was nevertheless the most adequate in terms of factor validity and proved invariant in independent samples. Convergent validity was only adequate for the factor "Search for Health Information", while discriminant validity was adequate for the factors "Search for Information" and "Understanding Information". Internal consistency showed adequate results on all the items. The second-order hierarchical model, although not totally adequate, slowly slightly higher fit indices and thus allowed calculating an overall health literacy score considering each item's best weight.


Increased vitamin D serum levels correlate with clinical improvement of rheumatic diseases after Dead Sea climatotherapy.
Ultraviolet B (UVB) rays are required by the skin for the production of vitamin D. The intensity of UVB at the Dead Sea area is the lowest in the world. Low vitamin D levels are often associated with musculoskeletal symptoms. To assess the effectiveness of climatotherapy at the Dead Sea on the production of vitamin D in Norwegian patients suffering from various rheumatic diseases and to investigate possible associations between increased vitamin D serum levels, musculoskeletal symptoms and disease severity. Sixty Norwegian patients who came to the Dead Sea area for 21 days of medical rehabilitation were divided into three groups according to their diagnosis: chronic pain syndromes, i.e., low back pain or fibromyalgia (Group 1, n=33); rheumatoid arthritis (Group 2, n=16); and osteoarthritis (Group 3, n=11). Serum 25-hydroxyvitamin D (25-OH-D) levels were determined at arrival and prior to departure. The treatment protocol included daily sun exposure (climatotherapy), bathing in the Dead Sea and mineral spring water (balneotherapy), mud applications and fitness classes. 25-OH-D serum levels increased significantly from 71.3 +/- 26.6 nM at arrival to 89.3 +/- 23.2 nM prior to departure (P < 0.001). Adjusted for the initial levels of pain (assessed by a visual analog scale) and disease severity, a direct correlation was observed between increased 25-OH-D serum levels and pain reduction (P = 0.012) and reduction of disease severity (P = 0.02). Climatotherapy at the Dead Sea induces significant changes in vitamin D. Increased 25-OH-D serum levels are associated with reduced musculoskeletal pain and disease severity.


SOX17 Inhibits Tumor Metastasis Via Wnt Signaling In Endometrial Cancer.
Endometrial cancer (EC) is the most common gynecological malignancy with high incidence of metastasis, while the mechanism of metastasis in EC is not clear. Immunohistochemistry and real-time PCR assays were used to assess expression of SOX17 in paraffin-embedded tissues from EC patients and in EC cells. The migration of EC cells was assessed by wound-healing and Transwell assays as well as in an in vitro study of nude mice. In addition, the expression of specific proteins was analyzed by Western blot. We observed that SOX17 expression levels were relatively high in stage I EC specimens, and were significantly correlated with the epithelial cadherin (E-cadherin) and β-catenin expression. Additionally, stage II EC patients whose specimens had relatively high SOX17 expression levels had better outcomes. Wound-healing and Transwell assays and in vivo murine experiments revealed that SOX17 inhibited EC cell migration. Meanwhile, SOX17 increased expression of E-cadherin and decreased expression of β-catenin and proteins in the Wnt signaling pathway. Moreover, LiCl (β-catenin activator) enhanced the regulatory effects of SOX17 on the expression of E-cadherin, promigratory cadherin, vimentin, and proteins in the Wnt signaling pathway, while XAV93920 (β-catenin inhibitor) exerted the opposite effect. The SOX17 N-terminus was proved to be necessary for these effects. Mechanistic investigations suggested SOX17 inhibits EC cell migration by inactivating the Wnt/β-catenin-epithelial mesenchymal transition (EMT) axis in EC cells. We uncovered a common SOX17-β-catenin-EMT mechanism underlying EC cell migration.


[Risk factors for coronary artery lesions in children with Kawasaki disease].
To study risk factors for the development of coronary artery lesions (CAL) in children with Kawasaki disease (KD). The clinical data of 527 children with KD between January 2006 and January 2009 were retrospectively reviewed. A total of 15 potential factors associated with occurrence of CAL were evaluated by univariate analysis and multivariate logistic regression analysis. The univariate analysis showed that age, gender, KD type, starting time of intravenous immunoglobulin (IVIG) treatment, response to IVIG treatment, additional treatment with corticosteroids, duration of fever and serum C-reactive protein level were significantly different between patients with and without CAL (P<0.05). Multivariate logistic regression analysis showed that an age of less than 1 year (OR=2.076, P<0.05) or greater than 8 years (OR=1.890, P<0.05), male sex (OR=1.972, P<0.05), incomplete KD (OR=1.426, P<0.05), delayed starting time of IVIG treatment (10 days after onset) (OR=3.251, P<0.05), no response to IVIG (OR=2.301, P<0.05) and fever duration of more than 10 days (OR=1.694, P<0.05) were independent risk factors for the development of CAL, whereas early starting time of IVIG treatment (before 5 days after onset) was a protective factor (OR=0.248, P<0.05). The occurrence of CAL is associated with many factors in children with KD. Age of less than 1 year or greater than 8 years, male sex, incomplete KD, delayed IVIG treatment after onset, no response to IVIG treatment and prolonged fever duration have been identified as risk factors for the development of CAL.


A workshop report on HIV mHealth synergy and strategy meeting to review emerging evidence-based mHealth interventions and develop a framework for scale-up of these interventions.
mHealth is a term used to refer to mobile technologies such as personal digital assistants and mobile phones for healthcare. mHealth initiatives to support care and treatment of patients are emerging globally and this workshop brought together researchers, policy makers, information, communication and technology programmers, academics and civil society representatives for one and a half days synergy meeting in Kenya to review regional evidence based mHealth research for HIV care and treatment, review mHealth technologies for adherence and retention interventions in anti-retroviral therapy (ART) programs and develop a framework for scale up of evidence based mHealth interventions. The workshop was held in May 2011 in Nairobi, Kenya and was funded by the Canadian Global Health Research Initiatives (GHRI) and the US Centre for Disease Control and Prevention (CDC). At the end of the workshop participants came up with a framework to guide mHealth initiatives in the region and a plan to work together in scaling up evidence based mHealth interventions. The participants acknowledged the importance of the meeting in setting the pace for strengthening and coordinating mHealth initiatives and unanimously agreed to hold a follow up meeting after three months.


Association of Increased Levels of lncRNA H19 in PBMCs with Risk of Coronary Artery Disease.
Considerable research shows that long non-coding RNAs, those longer than 200 nucleotides, are involved in several human diseases such as various cancers and cardiovascular diseases. Their significant role in regulating the function of endothelial cells, smooth muscle cells, macrophages, vascular inflammation, and metabolism indicates the possible effects of lncRNAs on the progression of atherosclerosis which is the most common underlying pathological process responsible for coronary artery disease (CAD). The aim of present study was to assess whether the expression of the lnc RNA H19 was associated with a susceptibility to CAD by evaluating the expression level of H19 in the peripheral blood. A case-control study of 50 CAD patients and 50 age and sex-matched healthy controls was undertaken to investigate whether the H19 lncRNA expression level is associated with a CAD using Taqman Real-Time polymerase chain reaction (PCR). The subsequent result indicated that the H19 lncRNA was over-expressed in CAD patients in comparison with the controls. However, it was not statistically significant. This overexpression may be involved in coronary artery disease progression. We report here, the up-regulation of H19 lncRNA in the whole blood of CAD patients and suggest a possible role for H19 in the atherosclerosis process and its consideration as novel biomarker for CAD.


'It's much more grey than black and white': clinical geneticists' views on the oversight of consumer genomics in Europe.
Aim: Direct-to-consumer (DTC) genetic tests (GT) have created controversy regarding their risks and benefits. In view of recent regulatory developments, this article aims to explore the attitudes of European clinical geneticists toward the oversight of DTC GT. Materials & methods: Fifteen semi-structured interviews were performed with clinical geneticists based in ten European countries. The transcripts were thematically analysized in an iterative process. Results & conclusion: Respondents strongly supported quality standards for DTC GT equal to those applied within the healthcare setting. Despite participants unanimously considering the involvement of healthcare professionals to be important, mandatory medical supervision was controversial. In this regard, promoting education and truth-in-advertising was considered as being key in maintaining a balance between protecting consumers and promoting their autonomy.


Monitoring genetic diversity with new indicators applied to an alpine freshwater top predator.
Genetic diversity is the basis for population adaptation and long-term survival, yet rarely considered in biodiversity monitoring. One key issue is the need for useful and straightforward indicators of genetic diversity. We monitored genetic diversity over 40 years (1970-2010) in metapopulations of brown trout (Salmo trutta) inhabiting 27 small mountain lakes representing 10 lake systems in central Sweden using >1200 fish per time point. We tested six newly proposed indicators; three were designed for broad, international use in the UN Convention on Biological Diversity (CBD) and are currently applied in several countries. The other three were recently elaborated for national use by a Swedish science-management effort and applied for the first time here. The Swedish indicators use molecular genetic data to monitor genetic diversity within and between populations (indicators ΔH and ΔFST , respectively) and assess the effective population size (Ne -indicator). We identified 29 genetically distinct populations, all retained over time. Twelve of the 27 lakes harboured more than one population indicating that brown trout biodiversity hidden as cryptic, sympatric populations are more common than recognized. The Ne indicator showed values below the threshold (Ne ≤ 500) in 20 populations with five showing Ne < 100. Statistically significant genetic diversity reductions occurred in several populations. Metapopulation structure appears to buffer against diversity loss; applying the indicators to metapopulations suggest mostly acceptable rates of change in all but one system. The CBD indicators agreed with the Swedish ones but provided less detail. All these indicators are appropriate for managers to initiate monitoring of genetic biodiversity.


Acanthamoeba keratitis. Potential role for topical clotrimazole in combination chemotherapy.
Clotrimazole is an antifungal agent that has been shown to have excellent in vitro activity against most strains of Acanthamoeba. We encountered four patients who developed Acanthamoeba keratitis while wearing contact lenses that had been stored in homemade saline. Their medical treatment regimens included the use of topical 1% clotrimazole. In two patients in whom conventional therapy failed, clotrimazole was successful in controlling recurrent infection following penetrating keratoplasty. Two other patients were treated with clotrimazole as well as propamidine isethionate and neomycin sulfate-polymyxin B sulfate-gramicidin from the outset, and had an excellent response to medical therapy. In those patients who found the commercially available cream uncomfortable, a 1% clotrimazole suspension formulated in artificial tears was used and found to be well tolerated.


Factors related to complications among adult patients with intellectual disabilities hospitalized at an academic medical center.
People with intellectual disabilities (ID) represent a small but important group of hospitalized patients who have higher rates of complications than do patients without ID hospitalized for the same reasons. Complications are potentially avoidable conditions, such as healthcare-acquired infections, healthcare-acquired skin breakdown, falls, and medication errors and reactions. Addressing factors related to complications can focus efforts to improve hospital care. The purpose of this exploratory study was to analyze data from reviews of academic medical center charts (N = 70) about complications and to examine patient and hospitalization characteristics in relation to complications among adult patients (age ≥ 18 years) with ID hospitalized for nonpsychiatric reasons. Adults with ID tended to be twice as likely to have complications (χ2 = 2.893, df = 1, p = .09) if they had a surgical procedure and were nearly four times as likely to have complications (χ2 = 6.836, df = 1, p = .009) if they had multiple chronic health conditions (three of the following: history of cerebral palsy, autism spectrum symptoms, aggressive behavior, respiratory disorder, and admission through the emergency department). Findings suggest preliminary criteria for assessing risk for complications among hospitalized people with ID and the need for attention to their specific needs when hospitalized.


The risk of acute pancreatitis associated with acid-suppressing drugs.
To assess the risk of acute pancreatitis associated with use of acid-suppressing drugs. We conducted a retrospective cohort study with a nested case-control design within the General Practice Research Database (GPRD) in the United Kingdom. The cohort included 180 178 persons aged 20-74 years, who had received at least one prescription of cimetidine, famotidine, nizatidine, ranitidine, lansoprazole, or omeprazole from January 1992 to September 1997 and who did not have major risk factors for pancreatic diseases. Patients with a computerized medical history compatible with idiopathic acute pancreatitis were validated through review of medical records. For the nested case-control analysis 1000 controls were randomly selected from the study population. We identified 88 potential cases of idiopathic acute pancreatitis. Medical records were available for 86. After review of these records 36 cases of acute pancreatitis were confirmed. Seven cases occurred during nonuse, corresponding to a background incidence rate (IR) of 4.4/100 000 person-years (PY). Six cases occurred during current use of ranitidine (IR 10.5/100 000 PY), five patients were current users of cimetidine (IR 13.9/100 000 PY), and three were current users of omeprazole (IR 7.8/100 000 PY). There were no cases among current users of famotidine, lansoprazole, or nizatidine. Relative risk (RR) compared with nonuse and corrected for age, gender, calendar year and use of medication known to be associated with acute pancreatitis was 1.3 (95% CI: 0.4,4.1) for ranitidine, 2.1 (95% CI: 0.6,7.2) for cimetidine, and 1.1 (95% CI: 0.3,4.6) for omeprazole. The results of this study do not support an association between acute pancreatitis and the use of acid-suppressing drugs, although a substantial increase in risk cannot be excluded with confidence.


Optogenetics for retinal disorders.
Optogenetics is the use of genetic methods combined with optical technology to achieve gain or loss of function within neuronal circuits. The field of optogenetics has been rapidly expanding in efforts to restore visual function to blinding diseases such as retinitis pigmentosa (RP). Most work in the field includes a group of light-sensitive retinaldehyde-binding proteins known as opsins. Opsins couple photon absorption to molecular signaling chains that control cellular ion currents. Targeting of opsin genes to surviving retinal cells is fundamental to the success of optogenetic therapy. Viral delivery, primarily adeno-associated virus, using intravitreal injection for inner retinal cells and subretinal injection for outer retinal cells, has proven successful in many models. Challenges in bioengineering remain for optogenetics including relative insensitivity of opsins to physiologic light levels of stimulation and difficulty with viral delivery in primate models. However, targeting optogenetic therapy may present an even greater challenge. Neural and glial remodeling seen in advanced stages of RP result in reorganization of remaining neural retina, and optogenetic therapy may not yield functional results. Remodeling also poses a challenge to the selection of cellular targets, with bipolar, amacrine and ganglion cells all playing distinct physiologic roles, and affected by remodeling differently. Although optogenetics has drawn closer to clinical utility, advances in opsin engineering, therapeutic targeting and ultimately in molecular inhibition of remodeling will play critical roles in the continued clinical advancement of optogenetic therapy.


Insulin-like growth factor I (IGF-I) receptors and IGF-I action in oligodendrocytes from rat brains.
Oligodendrocyte progenitor cells were prepared by mechanical dissociation of 1-day-old rat brain cultures. These cells undergo proliferation and differentiation into oligodendrocytes as demonstrated by the expression of proliferation and differentiation-related specific antigens. We have used this unique culture system to characterize insulin-like growth factor I (IGF-I) receptors and their action in the central nervous system (CNS). 125I-IGF-I specifically binds to these cultures with high affinity. Competition-inhibition data suggest that IGF-I is most potent in competing for 125I-IGF-I binding, followed by IGF-II and insulin. Scatchard analyses of the binding data indicate a curvilinear plot with a Kd for high affinity of 0.2 nM, and a Bmax of 247 fmol/mg, and a Kd for low affinity of 3.2 nM and Bmax of 1213 fmol/mg protein. Covalent cross-linking followed by SDS-PAGE analysis demonstrated a radioactive band of Mr 135,000 which corresponds to the alpha subunit of the IGF-I receptor. Solution hybridization/RNase protection assay produced a single protected band corresponding to IGF-I receptor messenger RNA, further confirming the presence of these receptors. Incubation of progenitor cells with IGF-I resulted in a time- and concentration-dependent increase in [3H]thymidine incorporation and cell numbers. This effect appears to be mediated by IGF-I receptors since IGF-II and insulin were proportionately less potent. In addition to its effect on proliferation, IGF-I also increased the number of 4E7- and GC-antigen positive cells. These observations indicate that oligodendrocytes in primary culture express specific IGF-I receptors and that the interaction of IGF-I with these receptors results in the proliferation as well as differentiation of oligodendrocytes.


Meningovertebral ligaments and their putative significance in low back pain.
To determine the presence and morphology of the meningovertebral ligaments (ligaments of Hofmann) as well as postulate their possible contribution to low back pain. Sagittal dissections were performed on 12 embalmed cadaver specimens including the L5/S1 intervertebral level cephalad to T1. Meningovertebral ligaments were labeled and documented in both the lumbar and thoracic regions. Meningovertebral ligaments were found in both the lumbar and thoracic regions of all cadaveric specimens. These ligaments were much more prevalent in the lumbar vertebral column but were also present throughout the thoracic vertebral column. The meningovertebral ligaments in the lumbar region were more robust as well as more frequently encountered than those found in the thoracic region. Dural sac attachments to the posterior aspect of the vertebral bodies and the posterior longitudinal ligament could act to traction the dural sac in the event of nuclear bulge or herniation. The prevalence of these ligaments in the lumbar spine, coupled with the high incidence of herniated nucleus pulposus and disc bulges in this region, may compound the effects of disc pathology and result in increased low back pain.


Computational Modeling of Molecular Structures Guided by Hydrogen-Exchange Data.
Data produced by hydrogen-exchange monitoring experiments have been used in structural studies of molecules for several decades. Despite uncertainties about the structural determinants of hydrogen exchange itself, such data have successfully helped guide the structural modeling of challenging molecular systems, such as membrane proteins or large macromolecular complexes. As hydrogen-exchange monitoring provides information on the dynamics of molecules in solution, it can complement other experimental techniques in so-called integrative modeling approaches. However, hydrogen-exchange data have often only been used to qualitatively assess molecular structures produced by computational modeling tools. In this paper, we look beyond qualitative approaches and survey the various paradigms under which hydrogen-exchange data have been used to quantitatively guide the computational modeling of molecular structures. Although numerous prediction models have been proposed to link molecular structure and hydrogen exchange, none of them has been widely accepted by the structural biology community. Here, we present as many hydrogen-exchange prediction models as we could find in the literature, with the aim of providing the first exhaustive list of its kind. From purely structure-based models to so-called fractional-population models or knowledge-based models, the field is quite vast. We aspire for this paper to become a resource for practitioners to gain a broader perspective on the field and guide research toward the definition of better prediction models. This will eventually improve synergies between hydrogen-exchange monitoring and molecular modeling.


Measuring and Controlling the Local Environment of Surface-Bound DNA in Self-Assembled Monolayers on Gold When Prepared Using Potential-Assisted Deposition.
DNA self-assembled monolayers (SAMs) were prepared using potential-assisted deposition on clean gold single-crystal bead electrodes under a number of conditions (constant or square-wave potential perturbations in TRIS or phosphate immobilization buffers with and without Cl-). The local environment around the fluorophore-labeled DNA tethered to the electrode surface was characterized using in situ fluorescence microscopy during electrochemical measurements as a function of the underlying surface crystallography. Potential-assisted deposition from a TRIS buffer containing Cl- created DNA SAMs that were uniformly distributed on the surface with little preference to the underlying crystallography. A constant (+0.4 V/SCE) or a square-wave potential perturbation (+0.4 to -0.3 V/SCE, 50 Hz) resulted in similar DNA-modified surfaces in TRIS immobilization buffer. Deposition using a square-wave potential without Cl- resulted in lower DNA surface coverage. Despite this, the local environment around the DNA in the SAM appears to be densely packed. This implies the formation of clusters of densely packed DNA in the SAM. This effect was also demonstrated when depositing from a phosphate buffer. DNA clusters were significantly reduced when Cl- was present in the buffer. Clusters were most prevalent on the low-index plane surfaces (e.g., {111} and {100}) and less on the higher-index planes (e.g., {210} or {311}). A mechanism is proposed to rationalize the formation of DNA-clustered regions for deposition using a square-wave potential perturbation. The conditions for creating clusters of DNA in a SAM or for preventing these clusters from forming provide an approach for tailoring the surfaces used for biosensing.


Estradiol effects on the growth hormone/insulin-like growth factor-1 axis in amenorrheic athletes.
Disruption of the growth hormone/insulin-like growth factor-1 (GH/IGF-1) axis has been reported and studied in menopause, hypothalamic amenorrhea, and anorexia nervosa, but not in weight-stable amenorrheic athletes. We investigated the effects of short-term transdermal estradiol on basal and exercise-stimulated serum GH, IGF-1, and associated binding proteins (IGFBP-1 and IGFBP-3) in seven weight-stable female amenorrheic athletes with percentage body fats greater that 12%. Each subject received a 72 h placebo patch followed by 144 h of transdermal estradiol. Serum samples for GH, IGF-1, IGFBP-1, and IGFBP-3 were obtained at baseline (t1), 72 hr (t2), 144 hr (t3), and during three 90-minute trials of aerobic exercise. Basal, and exercise GH, IGF-1, and IGFBP-1 were not different between trials. Baseline IGFBP-3 decreased from t1 to t2 (p = 0.04) and serum free fatty acids increased from t1 to t2, and t1 to t3 (p = 0.04, and 0.02 respectively). These findings differ from postmenopausal women, and women having weightloss-associated amenorrhea, suggesting that estrogen, exercise, and nutritional deficiencies may have independent effects on the GH/IGF-1 axis.


[Euthanasia and the abuse of conscience clause concept].
Since 2002, the Belgian legal framework authorizes the practice of euthanasia, under cer tain clear conditions. All cases have to be reported to the Assessment and Control Commission (ACC). To date, more than 10.000 cases have been reported since 2002. The concept of individual conscience clause is of concern and its use in an institutional dimension is analysed. The role of moral integrity and conscience clause in health care should be downplayed in the field of medicine's core values.


Langevin Approach to Quantum Optics with Molecules.
We investigate the interaction between light and molecular systems modeled as quantum emitters coupled to a multitude of vibrational modes via a Holstein-type interaction. We follow a quantum Langevin equations approach that allows for analytical derivations of absorption and fluorescence profiles of molecules driven by classical fields or coupled to quantized optical modes. We retrieve analytical expressions for the modification of the radiative emission branching ratio in the Purcell regime and for the asymmetric cavity transmission associated with dissipative cross talk between upper and lower polaritons in the strong coupling regime. We also characterize the Förster resonance energy transfer process between donor-acceptor molecules mediated by the vacuum or by a cavity mode.


Synthesis of novel pancreatic lipase inhibitors: Biological investigation and in silico studies.
The targeted compounds which are Schiff base derivatives were prepared by reaction of 6-(4-phenyl-piperazin-1-yl)pyridine-3-ylamine with 2-hydroxy and 2,6-dichloro benzaldehyde. These compounds were isolated, purified and then spectrally characterized via FT-IR, 1H and 13C NMR, LC MS TOF, and TGA analysis where strong proofs confirmed the formation of the targeted product. The biological activity, which is pancreatic porcine lipase inhibition, of the compounds was investigated and Orlistat was used as standard drug. The compound 3 was found to be as potent as orlistat against PL enzyme with an IC50 value of 0.50 μM. The molecular docking studies were performed for both obtained compounds and orlistat against active side of porcine pancreatic lipase. Also, MM/PBSA binding free energy and molecular dynamics (MD) simulation analyzes were performed for pancreatic porcine lipase with compound 3, which showed potent inhibition according to the results of in vitro studies. Furthermore, The ADME and toxicity analysis of the compounds were examined using web-based online platforms, SwissADME and pkCSM. In the light of biological activity and in silico studies, the compound 3 can be a potential drug candidate with further studies.Communicated by Ramaswamy H. Sarma.


Sediment toxicity identification evaluation (TIE phases I and II) based on microscale bioassays for diagnosing causes of toxicity in coastal areas affected by domestic sewage.
Domestic sewage is a major problem in highly urbanized coastal areas worldwide. In the present study, toxicity identification evaluation (TIE) phases I and II were applied to sediment interstitial water from 2 locations along the São Paulo coast in southeastern Brazil: the sewage outfalls from the city of Santos, a densely urbanized area, and the city of Bertioga, a less urbanized area. An adapted microscale sea urchin embryo-larval development bioassay was employed. Phase TIE-I manipulations were 1) ethylenediamine tetraacetic acid (EDTA) addition, 2) aeration at modified pH, 3) C18 solid-phase extraction (SPE), and 4) addition of Na2 S2 O3 . The results of the Santos TIE-I indicated toxicity by sulfides and substances with affinity to C18 resin. In Bertioga, toxicity was the result of NH3 , metals, and arsenic, as well as substances with affinity to C18 resin. Phase TIE-II aimed to specify the causes of toxicity by testing the toxicity of eluates of the sublation and C18 SPE manipulation. The results reinforced the role of both surfactants and nonpolar compounds as causative agents of toxicity in both Santos and Bertioga. Chemical analyses of sediment interstitial water or whole sediment ruled out the influence of polycyclic aromatic hydrocarbons (PAHs) or polychlorinated biphenyls (PCBs) in the toxicity of both sampling sites. Other hydrophobic substances may play a role in the toxicity of Santos and Bertioga effluents. Efforts to remove such substances from sanitary effluents must be prioritized. Environ Toxicol Chem 2017;36:1820-1832. © 2017 SETAC.


Spiral ganglion stem cells can be propagated and differentiated into neurons and glia.
The spiral ganglion is an essential functional component of the peripheral auditory system. Most types of hearing loss are associated with spiral ganglion cell degeneration which is irreversible due to the inner ear's lack of regenerative capacity. Recent studies revealed the existence of stem cells in the postnatal spiral ganglion, which gives rise to the hope that these cells might be useful for regenerative inner ear therapies. Here, we provide an in-depth analysis of sphere-forming stem cells isolated from the spiral ganglion of postnatal mice. We show that spiral ganglion spheres have characteristics similar to neurospheres isolated from the brain. Importantly, spiral ganglion sphere cells maintain their major stem cell characteristics after repeated propagation, which enables the culture of spheres for an extended period of time. In this work, we also demonstrate that differentiated sphere-derived cell populations not only adopt the immunophenotype of mature spiral ganglion cells but also develop distinct ultrastructural features of neurons and glial cells. Thus, our work provides further evidence that self-renewing spiral ganglion stem cells might serve as a promising source for the regeneration of lost auditory neurons.


Pharmacophore-Based Screening, Molecular Docking, and Dynamic Simulation of Fungal Metabolites as Inhibitors of Multi-Targets in Neurodegenerative Disorders.
Neurodegenerative disorders, such as Alzheimer's disease (AD), negatively affect the economic and psychological system. For AD, there is still a lack of disease-altering treatments and promising cures due to its complex pathophysiology. In this study, we computationally screened the natural database of fungal metabolites against three known therapeutic target proteins of AD. Initially, a pharmacophore-based, drug-likeness category was employed for screening, and it filtered the 14 (A-N) best hits out of 17,544 fungal metabolites. The 14 best hits were docked individually against GSK-3β, the NMDA receptor, and BACE-1 to investigate the potential of finding a multitarget inhibitor. We found that compounds B, F, and L were immuno-toxic, whereas E, H, I, and J had a higher LD50 dose (5000 mg/kg). Among the examined metabolites, the Bisacremine-C (compound I) was found to be the most active molecule against GSK-3β (ΔG: -8.7 ± 0.2 Kcal/mol, Ki: 2.4 × 106 M-1), NMDA (ΔG: -9.5 ± 0.1 Kcal/mol, Ki: 9.2 × 106 M-1), and BACE-1 (ΔG: -9.1 ± 0.2 Kcal/mol, Ki: 4.7 × 106 M-1). It showed a 25-fold higher affinity with GSK-3β, 6.3-fold higher affinity with NMDA, and 9.04-fold higher affinity with BACE-1 than their native ligands, respectively. Molecular dynamic simulation parameters, such as RMSD, RMSF, Rg, and SASA, all confirmed that the overall structures of the targeted enzymes did not change significantly after binding with Bisacremine-C, and the ligand remained inside the binding cavity in a stable conformation for most of the simulation time. The most significant hydrophobic contacts for the GSK-3β-Bisacremine-C complex are with ILE62, VAL70, ALA83, and LEU188, whereas GLN185 is significant for H-bonds. In terms of hydrophobic contacts, TYR184 and PHE246 are the most important, while SER180 is vital for H-bonds in NMDA-Bisacremine-C. THR232 is the most crucial for H-bonds in BACE-1-Bisacremine-C and ILE110-produced hydrophobic contacts. This study laid a foundation for further experimental validation and clinical trials regarding the biopotency of Bisacremine-C.


Explaining sudden infant death with cardiac arrhythmias: Complete exon sequencing of nine cardiac arrhythmia genes in Dutch SIDS cases highlights new and known DNA variants.
Previous studies suggested that Sudden Infant Death Syndrome (SIDS) can partially be genetically explained by cardiac arrhythmias; however, the number of individuals and populations investigated remain limited. We report the first SIDS study on cardiac arrhythmias genes from the Netherlands, a country with the lowest SIDS incidence likely due to parent education on awareness of environmental risk factors. By using targeted massively parallel sequencing (MPS) in 142 Dutch SIDS cases, we performed a complete exon screening of all 173 exons from 9 cardiac arrhythmias genes SCN5A, KCNQ1, KCNH2, KCNE1, KCNE2, CACNA1C, CAV3, ANK2 and KCNJ2 (∼34,000 base pairs), that were selected to harbour previously established SIDS-associated DNA variants. Motivated by the poor DNA quality from the paraffin embedded material used, the application of a conservative sequencing quality control protocol resulted in 102 SIDS cases surviving quality control. Amongst the 102 SIDS cases, we identified a total of 40 DNA variants in 8 cardiac arrhythmia genes found in 60 (58.8 %) cases. Statistical analyses using ancestry-adjusted reference population data and multiple test correction revealed that 13 (32.5 %) of the identified DNA variants in 6 cardiac arrhythmia genes were significantly associated with SIDS, which were observed in 15 (14.7 %) SIDS cases. These 13, and another three, DNA variants were classified as likely pathogenic for cardiac arrhythmias using the American College of Medical Genetics guidelines for interpretation of sequence variants. The 16 likely pathogenic DNA variants were found in 16 (15.7 %) SIDS cases, including i) 3 novel DNA variants not recorded in public databases ii) 7 known DNA variants for which significant SIDS association established here was previously unknown, and iii) 6 known DNA variants for which LQTS association was reported previously. By having replicated previously reported SIDS-associated DNA variants located in cardiac arrhythmia genes and by having highlighting novel SIDS-associated DNA variants in such genes, our findings provide additional empirical evidence for the partial genetic explanation of SIDS by cardiac arrhythmias. On a wider note, our study outcome stresses the need for routine post-mortem genetic screening of assumed SIDS cases, particularly for cardiac arrhythmia genes. When put in practise, it will allow preventing further sudden deaths (not only in infants) in the affected families, thereby allowing forensic molecular autopsy not only to provide answers on the cause of death, but moreover to save lives.


Local Fitting of the Kohn-Sham Density in a Gaussian and Plane Waves Scheme for Large-Scale Density Functional Theory Simulations.
A local resolution-of-the-identity (LRI) approach is introduced in combination with the Gaussian and plane waves (GPW) scheme to enable large-scale Kohn-Sham density functional theory calculations. In GPW, the computational bottleneck is typically the description of the total charge density on real-space grids. Introducing the LRI approximation, the linear scaling of the GPW approach with respect to system size is retained, while the prefactor for the grid operations is reduced. The density fitting is an O(N) scaling process implemented by approximating the atomic pair densities by an expansion in one-center fit functions. The computational cost for the grid-based operations becomes negligible in LRIGPW. The self-consistent field iteration is up to 30 times faster for periodic systems dependent on the symmetry of the simulation cell and on the density of grid points. However, due to the overhead introduced by the local density fitting, single point calculations and complete molecular dynamics steps, including the calculation of the forces, are effectively accelerated by up to a factor of ∼10. The accuracy of LRIGPW is assessed for different systems and properties, showing that total energies, reaction energies, intramolecular and intermolecular structure parameters are well reproduced. LRIGPW yields also high quality results for extended condensed phase systems such as liquid water, ice XV, and molecular crystals.


Dual targeting agents for Aβ plaque/P-glycoprotein and Aβ plaque/nicotinic acetylcholine α4β2* receptors-potential approaches to facilitate Aβ plaque removal in Alzheimer's disease brain.
Alzheimer's disease (AD) affects 10% of people older than 65 and is characterized by a progressive loss of cognitive function with an abnormal accumulation of amyloid β (Aβ ) plaques and neurofibrillary tangles (NFT) in the brain. Efforts to reduce brain Aβ plaques continue to be investigated as a therapeutic approach for AD. We report here development of dual targeting agents with affinity for Aβ plaque/P-glycoprotein (Pgp) and Aβ plaque/α4β 2* nicotinic acetylcholine receptors (nAChR). These novel dual agents may be able to efflux Aβ plaques via the paravascular (glymphatic) pathways. Ferulic acid (FA), ferulic acid ethyl ester (FAEE), and curcumin (CUR) were used for Aβ plaques, fexofenadine (FEX) was used as substrate for Pgp and nifrolidine (NIF) was used for α4β 2* nAChRs. Aβ plaque/α4β 2* nAChR dual agent, FA-NIF (GKS-007) exhibited IC50 = 3-6 nM for α4β 2* nAChRs in [3H]cytisine-radiolabeled thalamus and frontal cortex in rat brain slices. In postmortem human AD frontal cortex, Aβ plaques labeled with [3H]PIB, FEX-CUR showed a 35% reduction in gray matter (GM)/white matter (WM) [3H]PIB binding, while CUR alone showed a 50% reduction. In vivo biodistribution studies are required of the Aβ-Pgp and Aβ-α4β 2* nAChRs dual targeting agents in order to evaluate their potential as therapeutic approaches for reducing brain Aβ plaques.


Nefazodone in the adjunctive therapy of schizophrenia: an open-label exploratory study.
Compared to conventional antipsychotic medications, atypical antipsychotic medications demonstrate greater central serotonin (5HT2) receptor antagonism than dopamine type 2 (D2) receptor antagonism. Nefazodone, an antidepressant medication, exhibits 5HT2 receptor antagonism; we therefore wondered if its addition to stable regimens of antipsychotic medication would increase antipsychotic efficacy, independently of a primary effect on mood, through the mechanism of augmented 5HT2 receptor antagonism. In a pilot investigation, we administered nefazodone (400 mg/d) for 6 weeks as an open-label adjunct to antipsychotic medication in 10 patients with chronic schizophrenia. The patients were moderately depressed at baseline but did not meet criteria for major depressive episode. The Brief Psychiatric Rating Scale (BPRS) and Montgomery-Asberg Depression Rating Scale scores showed statistically significant and clinically robust improvements with nefazodone treatment, which were maintained at follow-up evaluation 2 weeks after the end of nefazodone treatment. There were no adverse events. These results suggest that nefazodone may be a safe and effective adjunct to antipsychotic medications in schizophrenia and that augmentation of 5HT2 antagonism may prove to be a viable strategy for "boosting" antipsychotic efficacy and for treating depressive symptoms in schizophrenia.


Evaluation of regional limb perfusion with erythromycin using the saphenous, cephalic, or palmar digital veins in standing horses.
There are no reported studies evaluating the use of erythromycin for regional limb perfusion (RLP) in horses. Our hypothesis was that using the cephalic and saphenous veins for RLP will enable delivery of therapeutic concentrations of erythromycin to the distal limb. Nineteen healthy horses participated in the study. The cephalic, saphenous or palmar digital (PD) vein was used to perfuse the limb with erythromycin. Synovial samples were collected from the metacarpo/metatarso-phalangeal (MCP/MTP) joint and blood samples were collected from the jugular vein. Maximum concentration (C(max)) of erythromycin in the MCP joint using the cephalic vein was 113 mg/L. The Cmax of erythromycin in the MTP joint using the saphenous vein was 38 mg/L. Erythromycin administered using the PD vein was not detectable in the MCP/MTP joint of four of six horses. Concentrations of erythromycin achieved in the synovial fluid of the MCP/MTP joint were between 152 and 452 times the minimal inhibitory concentration (MIC) for Rhodococcus equi (R. equi). In conclusion, the results indicate that when using the saphenous or cephalic veins for RLP, therapeutic concentrations of erythromycin in the MCP/MTP joint can be consistently reached [corrected].


Synthesis and in vivo evaluation of a radiofluorinated ketone body derivative.
The ketone bodies d-beta-hydroxybutyric acid and acetoacetic acid represent the principal oxidative energy sources of most tissues when dietary glucose is scarce. An 18F-labeled ketone body could be a useful tool for studying ketone body metabolism using positron emission tomography (PET). Here, we report the first radiofluorinated ketone body derivative (3S)-4-[18F]fluoro-3-hydroxybutyric acid ([18F]FBHB) as well as its enantiomer and l-beta-hydroxybutyric acid derivative, (3R)-4-[18F]fluoro-3-hydroxybutyric acid ((R)-[18F]F3HB). PET imaging in mice showed biodistribution profiles of the radiotracers that were consistent with the biodistribution of the respective endogenous compounds. Moreover, both enantiomers visualized breast cancer xenografts in vivo. Fasting over 24 h showed significantly enhanced brain and heart uptake of [18F]FBHB and tumor uptake of (R)-[18F]F3HB. Disorders exhibiting altered energy substrate utilization, such as Alzheimer's disease, epilepsy, diabetes, and cancer may be of interest for PET imaging studies using [18F]FBHB.


Ortho-substituted polychlorinated biphenyls alter calcium regulation by a ryanodine receptor-mediated mechanism: structural specificity toward skeletal- and cardiac-type microsomal calcium release channels.
We investigated a novel molecular mechanism by which polychlorinated biphenyls (PCBs) alter microsomal Ca2+ transport with sarcoplasmic reticulum (SR) membranes isolated from skeletal and cardiac muscles. Aroclors with an intermediate weight percent of chlorine enhance by >6-fold the binding of 1 nM[3H]ryanodine to its conformationally sensitive site on the SR Ca2+ -release channel [i.e., ryanodine receptor (RyR)] with high potency (EC50=1.4 microM), whereas Aroclors with either high or low chlorine composition show little activity. Structure-activity studies with selected pentachlorobiphenyl congeners reveal a stringent structural requirement for chlorine substitution at the ortho-positions, with 2,2',3,5',6-pentachlorobiphenyl having the highest potency toward skeletal and cardiac isoforms of RyR (EC50=330 nM and 2 microM, respectively). In contrast, 3,3',4,4',5-pentachlorobiphenyl does not enhance ryanodine binding, suggesting that noncoplanarity of the biphenyl rings is required for channel activation. However, 2,2',4,6,6'-pentachlorobiphenyl is significantly less active toward RyR, suggesting that some degree of rotation about the biphenyl bond is required. 2,2',3,5',6-Pentachlorobiphenyl induces a dose-dependent release of Ca2+ from actively loaded SR vesicles with a maximum rate of 1.2 micromol mg-1 min-1 (EC50=1 microM), whereas 3,3',4,4',5-pentachlorobiphenyl (< / = microM) does not alter Ca2+ transport. The mechanism of PCB-induced channel activation involves a significant decrease in the inhibitory potency of Ca2+ and Mg2+ (20-fold and 100-fold, respectively). Neither 2,2',3,5',6- nor 3,3',4,4',5-pentachlorobiphenyl (< / = 10 microM) alters the activity of the skeletal isoform of sarcoplasmic/endoplasmic reticulum Ca2+ -ATPase or the cardiac isoform of sarcoplasmic/endoplasmic reticulum Ca2+ -ATPase, and PCB-induced Ca2+ release can be fully blocked by either microM ryanodine or ruthenium red. These results are the first to demonstrate a selective ryanodine receptor-mediated mechanism by which ortho-substituted PCBs alter microsomal Ca2+ transport and may have toxicological relevance.


Characteristics of fast-food/takeaway-food and restaurant/café-food consumers among New Zealand adults.
To investigate: (i) the percentage of the New Zealand (NZ) population reporting fast food/takeaway food and restaurant/café food per day; (ii) examine demographic factors associated with their use; (iii) quantify their contribution to energy intake; and (iv) describe the specific types of foods reported from both sources. Twenty-four hour diet recalls from the cross-sectional 2008/09 NZ Adult Nutrition Survey were used to identify fast-food and restaurant-food consumers. NZ households. Adults aged 15 years and older (n 4721). Overall 28 % reported consuming at least one fast food and 14 % a restaurant food within the 24 h diet recall. Fast-food consumption was not associated with level of education or an area-based measure of socio-economic status, but a higher education was positively associated with restaurant-food consumption. Individual factors such as ethnicity, household size, age, sex and marital status were found to be important influences on the use of fast food and restaurant food. Fast-food consumption was more prevalent among participants living in urban areas, young adults (19-30 years) and Māori compared with NZ European and Others. The most frequently reported fast foods were bread-based dishes, potatoes (including fries) and non-alcoholic beverages. Given the high reported consumption of fast food by young adults, health promotion initiatives both to improve the nutritional quality of fast-food menus and to encourage healthier food choices would likely make a large impact on the overall diet quality of this group.


Transdermal clonidine in mild hypertension. A randomized, double-blind, placebo-controlled trial.
In 30 patients with mild essential hypertension, clonidine hydrochloride was delivered from a skin patch reservoir designed to release medication at a constant rate for seven days. After a four-week washout period, patients were randomized (double-blind) into a clonidine- or a placebo-treated group. Clonidine or placebo was then given for five weeks, followed by a two-week washout period to assess withdrawal from treatment. Blood pressure was controlled in 11 of 15 clonidine-treated patients but in only four of 15 placebo-treated patients. The clonidine-treated group evidenced larger decreases in both systolic and diastolic blood pressures. In the clonidine-treated group, blood pressures and plasma clonidine levels were stable throughout a representative seven-day period. Besides mild skin irritation with both clonidine and placebo patches, few side effects were observed. After discontinuation of clonidine administration, plasma levels declined in a non-log linear manner. There was no rebound hypertension. The results suggest that clonidine delivered transdermally is safe and effective for control of mild essential hypertension.


In vitro and pilot in vivo imaging of 18 kDa translocator protein (TSPO) in inflammatory vascular disease.
Inflammatory vascular disease of the arteries, such as inflamed atheromatous plaques or arteritis, may cause aneurysms or ischemic strokes. In this context, using positron emission tomography (PET) to image inflammation may help select patients who would benefit from appropriate therapeutic interventions. This study sought to assess the usefulness of the 18 kDa translocator protein (TSPO) tracers [11C]-PBR28 and [18F]-PBR06 for imaging inflammatory vascular disease in vitro and in vivo. Immunohistochemistry for macrophage infiltration as well as autoradiography with [18F]-PBR06 were performed on eight paraffin-embedded, formalin-fixed atherosclerosis plaques prospectively collected after carotid endarterectomy of eight patients affected by ischemic stroke. Six different patients, one of whom was also included in the in vitro study, underwent PET imaging. Two patients with carotid stenosis associated with ischemic stroke were imaged with [18F]-PBR06 PET/CT, and four other patients (three with large vessel vasculitis and one with bilateral carotid stenosis but without stroke) were imaged with [11C]-PBR28. All in vitro sections showed specific binding of [18F]-PBR06, which co-localized with immunohistochemistry markers for inflammation. However, in vivo TSPO imaging with either [11C]-PBR28 or [18F]-PBR06 was negative in all participants. Despite good uptake on surgical samples in vitro, [11C]-PBR28 and [18F]-PBR06 are not viable clinical tools for imaging inflammatory vascular disease. NCT02513589, registered 31 July 2015 and NCT00547976, registered 23 October 2007. https://clinicaltrials.gov .