Efficient Expression of HIV in Immunocompetent Mouse Brain Reveals a Novel Nonneurotoxic Viral Function in Hippocampal Synaptodendritic Injury and Memory Impairment.
HIV causes neurodegeneration and dementia in AIDS patients, but its function in milder cognitive impairments in virologically suppressed patients on antiretroviral therapy is unknown. Such patients are immunocompetent, have low peripheral and brain HIV burdens, and show minimal brain neuropathology. Using the model of HIV-related memory impairment in EcoHIV-infected conventional mice, we investigated the neurobiological and cognitive consequences of efficient EcoHIV expression in the mouse brain after intracerebral infection. HIV integrated and persisted in an expressed state in brain tissue, was detectable in brain monocytic cells, and caused neuroinflammatory responses and lasting spatial, working, and associative memory impairment. Systemic antiretroviral treatment prevented direct brain infection and memory dysfunction indicating the requirement for HIV expression in the brain for disease. Similarly inoculated murine leukemia virus used as a control replicated in mouse brain but not in monocytic cells and was cognitively benign, linking the disease to HIV-specific functions. Memory impairment correlated in real time with hippocampal dysfunction shown by defective long-term potentiation in hippocampal slices ex vivo and with diffuse synaptodendritic injury in the hippocampus reflected in significant reduction in microtubule-associated protein 2 and synapsin II staining. In contrast, there was no evidence of overt neuronal loss in this region as determined by neuron-specific nuclear protein quantification, TUNEL assay, and histological observations. Our results reveal a novel capacity of HIV to induce neuronal dysfunction and memory impairment independent of neurotoxicity, distinct from the neurotoxicity of HIV infection in dementia.IMPORTANCE HIV neuropathogenesis has been attributed in large measure to neurotoxicity of viral proteins and inflammatory factors produced by infected monocytic cells in the brain. We show here that HIV expression in mouse brain causes lasting memory impairment by a mechanism involving injury to hippocampal synaptodendritic arbors and neuronal function but not overt neuronal loss in the region. Our results mirror the observation of minimal neurodegeneration in cognitively impaired HIV patients on antiretroviral therapy and demonstrate that HIV is nonneurotoxic in certain brain abnormalities that it causes. If neurons comprising the cognition-related networks survive HIV insult, at least for some time, there is a window of opportunity for disease treatment.


The regulation of trophoblast differentiation by oxygen in the first trimester of pregnancy.
In the first trimester of human pregnancy villous cytotrophoblasts are able to differentiate to form either the overlying syncytiotrophoblast layer or, in anchoring villi, extravillous trophoblasts which grow out from the villi and invade into the maternal decidua, acting to both physically attach the placenta to the decidua, and modify the maternal spiral arteries to sustain pregnancy. During the first 10-12 weeks of gestation, extravillous trophoblast plugs block the spiral arteries and prevent maternal blood flow entering the intervillous space, thereby creating an environment of physiological hypoxia in which placental and fetal development occur. As extravillous trophoblasts migrate away from the villus they differentiate from a proliferative to an invasive phenotype. The hypoxic environment of the first trimester is believed to play an important role in the regulation of trophoblast differentiation. However, there is currently a large body of conflicting experimental evidence concerning this topic. This review examines the experimental evidence to date on the role of oxygen in trophoblast differentiation.


Effects of dietary energy levels on physiological parameters and reproductive performance of gestating sows over three consecutive parities.
This experiment was to evaluate the effects of the dietary energy levels on the physiological parameters and reproductive performance during gestation over three parities in sows. A total of 52 F1 gilts (Yorkshire×Landrace) were allotted to one of four dietary treatments using a completely randomized design. The treatments contained 3,100, 3,200, 3,300, or 3,400 kcal of metabolizable energy (ME)/kg diet but feed was provided at 2.0, 2.2, and 2.4 kg/d in the first, second and third parity, respectively. The body weight and body weight gain during gestation increased as the dietary energy level increased (p<0.05, and p<0.01) in the first parity. In the second parity, the body weight of sows was the lowest (p<0.05) when 3,100 kcal of ME/kg treatment diet was provided. The body weight was higher as the dietary energy level increased (p<0.05) during the gestation period in the third parity. During lactation, the voluntary feed intake of lactating sows tended to decrease when gilts were fed higher energy treatment diet (p = 0.08) and the body weight, body weight gain were increased by dietary energy level during gestation (p< 0.05). Backfat thickness was not affected by dietary treatment during the gestation period in three parities, interestingly backfat change from breeding to d 110 of gestation was higher as the dietary energy level increased at the first parity (p<0.05). When gilts were fed 3,400 kcal of ME/kg treatment diet a higher number of weaning piglets was observed in the first parity (p<0.05). The highest culling rate (69%) was seen when gestating sows were fed 3,100 kcal/kg ME treatment diet during three parities. In conclusion, the adequate energy intake of gestating sows should be 6,400 or 6,600 kcal of ME/d, 7,040 or 7,260 kcal of ME/d, and 7,680 or 7,920 kcal of ME/d for parity 1, 2, and 3, respectively.


Amitriptyline-induced cutaneous hyperpigmentation: case report and review of psychotropic drug-associated mucocutaneous hyperpigmentation.
Several drugs can be associated with hyperpigmentation of mucosa or skin. They include antibiotic, antimalarial, antineoplastic, and psychotropic medications. To describe a 42-year-old woman with amitriptyline-associated photo-distributed hyperpigmentation and to review psychotropic drug-induced hyperpigmentation of the skin. The features of a woman with amitriptyline-induced hyperpigmentation are presented. Using PubMed, the following terms were searched and relevant citations were assessed and discussed for context: amitriptyline, chlorpromazine, citalopram, desipramine, drug-associated, drug-induced, Fontana Masson, hyperpigmentation, imipramine, melanin, melanophages, mirtazapine, phenytoin, psychotropic, sertraline, thioridazine, tricyclic antidepressant. Photo-distributed hyperpigmentation on the upper back of a woman developed six and a half years after initiation of amitriptyline therapy. Biopsy of the affected area showed pigment-laden melanophages and intradermal melanin deposition. Psychotropic drugs associated with cutaneous hyperpigmentation include amitriptyline, chlorpromazine, citalopram, desipramine, imipramine, mirtazapine, phenytoin, sertraline, and thioridazine. The hyperpigmentation may initially appear many years after starting the medication. Pathology typically shows melanophages and melanin in the dermis. Fontana Masson stain confirms the presence of melanin; Perl stain for hemosiderin or iron is negative. Discontinuation of the drug may result in spontaneous improvement. Further studies are needed to better understand the role of Q-switched laser in treating drug-induced hyperpigmentation.


Near-infrared photothermal liposomal nanoantagonists for amplified cancer photodynamic therapy.
Photodynamic therapy (PDT) has been demonstrated to be a promising strategy for the treatment of cancer, while its therapeutic efficacy is often compromised due to excessive concentrations of glutathione (GSH) as a reactive oxygen species (ROS) scavenger in cancer cells. Herein, we report the development of near-infrared (NIR) photothermal liposomal nanoantagonists (PLNAs) for amplified PDT through through the reduction of intracellular GSH biosynthesis. Such PLNAs were constructed via encapsulating a photosensitizer, indocyanine green (ICG) and a GSH synthesis antagonist, l-buthionine sulfoximine (BSO) into a thermal responsive liposome. Under NIR laser irradiation at 808 nm, PLNAs generate mild heat via a ICG-mediated photothermal conversion effect, which leads to the destruction of thermal responsive liposomes for a controlled release of BSO in a tumor microenvironment, ultimately reducing GSH levels. This amplifies intracellular oxidative stresses and thus synergizes with PDT to afford an enhanced therapeutic efficacy. Both in vitro and in vivo data verify that PLNA-mediated phototherapy has an at least 2-fold higher efficacy in killing cancer cells and inhibiting tumor growth compared to sole PDT. This study thus demonstrates a NIR photothermal drug delivery nanosystem for amplified photomedicine.


TLR activation of the transcription factor XBP1 regulates innate immune responses in macrophages.
Sensors of pathogens, such as Toll-like receptors (TLRs), detect microbes to activate transcriptional programs that orchestrate adaptive responses to specific insults. Here we report that TLR4 and TLR2 specifically activated the endoplasmic reticulum (ER) stress sensor kinase IRE1alpha and its downstream target, the transcription factor XBP1. Previously described ER-stress target genes of XBP1 were not induced by TLR signaling. Instead, TLR-activated XBP1 was required for optimal and sustained production of proinflammatory cytokines in macrophages. Consistent with that finding, activation of IRE1alpha by ER stress acted in synergy with TLR activation for cytokine production. Moreover, XBP1 deficiency resulted in a much greater bacterial burden in mice infected with the TLR2-activating human intracellular pathogen Francisella tularensis. Our findings identify an unsuspected critical function for XBP1 in mammalian host defenses.


Disseminated Infection by Scedosporium/Lomentospora During Induction Therapy for Acute Myeloid Leukemia Complicated by Nontuberculous Mycobacteria.
Scedosporium/Lomentospora infections are rare and are associated with a high mortality rate in immunocompromised patients. A 69-year-old man with nontuberculous mycobacteria (NTM) died during induction chemotherapy for acute myeloid leukemia because of multiple organ failure due to pneumonia. During an autopsy, Lomentospora prolificans was detected using a fungal gene analysis of the blood, lungs, spleen, kidneys, and intestines, and Scedosporium aurantiacum was detected in the lungs. NTM disease may predispose patients to Scedosporium/Lomentospora infections. Physicians should consider Scedosporium/Lomentospora spp. as an invasive fungal infection that occurs during myelosuppression, particularly when NTM is a complication.


Visual evoked potentials in primary hypertension.
Functional integrity of sensory pathways in the brain has not been well documented in hypertension. It is suggested that vascular endothelial changes including hyalinisation during hypertension may lead to demyelination in the vulnerable areas of the brain. Since optic nerve is considered to be part of brain hence the present study was done to find out if visual pathways are involved in hypertension. Transient pattern reversal visual evoked potentials (VEPs) from 01, 02 scalp regions were recorded in 23 primary hypertensive patients and compared with 14 normotensive control. Of these, six patients showed delayed P1 latencies beyond 99% tolerance limit i.e. Mean + 3 SD of normal. The remaining 17 had latencies of all positive (P1-P3) and negative (N1-N3) waves comparable to those of the control group. Correlation Coefficient worked out, showed significant correlation between systolic BP and P1 latency in the control group only. No other parameter showed any correlation with P1 latency & amplitude in both the groups. These findings show that fluctuations of BP in normotensive subjects have correlation with P1 latency. This correlation ceased to exist in hypertensive patients and abnormality in P1 latency of VEP was detected in 26% cases.


Adolescent sources of stress, stress levels, and associations between stress and changes in weight outcomes at the time of COVID-19 related school closure in March 2020.
Stress is associated with weight changes, yet how level and sources of stress relate to this association is poorly understood. This mixed methods study examined associations between adolescent stress at the COVID-19 pandemic onset and standardized BMI (BMIz) over a three-month period. Methods: Participants (N = 197, mean age 13.66 ± 1.43 years, 85% Hispanic, 56% female) were recruited from a charter school in Texas to participate in a healthy lifestyle intervention during physical education class. We calculated BMIz using height and weight measurements taken December 2019 and the week of COVID-19 school closures in March 2020. We measured stress in March 2020 with the 4-item Perceived Stress Scale. We ran a multiple linear regression model controlling for baseline BMIz (December 2019), age, sex, intervention condition, and assessed the interaction between baseline BMIz and PSS-4 score on BMIz change. Overall, our sample had an average PSS-4 score of seven and mean BMIz decrease of 0.04 over the three-month period. We observed a significant interaction between PSS-4 score and baseline BMIz. At lower baseline BMIz scores, higher PSS-4 scores were associated with decreases in BMIz over time. There was no significant association at higher baseline BMIz scores. COVID-19 pandemic-related concerns and school performance were the two primary stressors reported. The relation between stress and weight changes differed based on starting weight status. Future interventions should consider the influence of external stressors on intervention adherence and outcomes.


Automated liver elasticity calculation for MR elastography.
MR elastography (MRE) is a phase-contrast MRI technique that is used to quantitatively assess liver stiffness for staging hepatic fibrosis. The current approach requires manual selection of a region of interest (ROI) with good wave quality from which to measure stiffness. The purpose of this work was to develop and evaluate a fully automated approach for measuring hepatic stiffness from MRE images to further reduce measurement variability. An automated liver elasticity calculation (ALEC) algorithm was developed to address reader stiffness measurement variability. ALEC has three stages: initial tissue estimation, segmentation, and ROI cleanup. Stiffnesses measured by the algorithm were compared with technicians and an expert radiologist in a set of 121 clinical cases acquired at 1.5 Tesla. Intra-class correlation (ICC), Bland-Altman analysis, and a noninferiority test were performed to evaluate whether the algorithm can be used in place of manual analysis by technicians. The stiffness measurement difference with the expert was 1.42% ± 11.17% (mean ± standard deviation) for the algorithm and 1.82% ± 13.65% for the technicians. The ICCs were 0.981 and 0.984, respectively. Both the algorithm and technicians were equivalent to the expert within a 5% significance margin (P < 0.01). The algorithm had no failures in the 119 cases that were considered analyzable by the human readers. The results of this study show that the newly developed automated algorithm is able to measure stiffness in clinical liver MRE exams with an accuracy that is equivalent to that of an expert radiologist. ALEC may be useful for analysis of archived data and suitable for performing multi-center studies.


Ablation of otolithic organs with argon laser.
Utricular maculae of guinea pigs were irradiated by argon laser through the oval window without touching the perilymph. The stapes was replaced in the oval window after irradiation. The inner ear was studied morphologically and physiologically. Argon laser beam caused detachment of the sensory epithelium from the basement membrane of the utricular macula. The lesion was round and localized at the site of irradiation. The epithelium changed to a layer of cuboidal cells after 2 to 3 weeks at the irradiated spot. Degeneration of the epithelium gradually spread to the periphery, and total loss of sensory epithelium was observed 10 weeks after irradiation. Hearing was preserved, and caloric test showed normal response.


The phosphorylation of the two-chain form of vitronectin by protein kinase A is heparin dependent.
In circulating blood, vitronectin occurs in two forms: a single-chain (75 kDa) and an endogenously clipped two-chain form (65 kDa and 10 kDa) held together by a disulfide bridge. The 75 kDa form was previously shown to be phosphorylated at Ser378 by protein kinase A, released by physiologically stimulated platelets. By contrast, at pH 7.5 the two-chain form is not phosphorylated at all. Heparin or heparan sulfate are shown here to modulate the conformation of clipped vitronectin at physiological pH, exposing Ser378 and allowing its stoichiometric phosphorylation by the kinase. At this pH the two-chain form of vitronectin in plasma exhibits a higher affinity for heparin, and behaves as a flexible molecule, which can conformationally respond to heparin and heparan sulfate, effectors involved in vitronectin function.


Relapsing pityriasis rosea.
To assess the prevalence of relapses of pityriasis rosea (PR), a retrospective cohort study investigated all PR cases diagnosed in Genoa between 2000 and 2013 and followed them up to today. Of 570 cases, 21 (3.7%) relapsed. Most of them had a single episode, but 4 had two episodes and 2 had three episodes. The herald patch was always absent, size and number of the lesions were reduced, and duration was shorter than that of the primary episodes. Constitutional symptoms were present, though less severe than in the primary eruption. Most recurrences occurred within 1 year (16/21, 76.2%). Men outnumbered women and the mean age of the relapsing patients (20.3 years) was higher than that for the primary episode. A pathogenetic hypothesis is provided: since PR is associated with reactivation of human herpesvirus 6/7, a parallelism with other typical reactivating human herpesviruses (varicella zoster virus and Epstein-Barr virus) has been established.


Illness understanding, causal attributions and emotional reactions in wives of myocardial infarction patients.
This study addresses lay illness understanding and attributions, including attribution of blame, in wives of myocardial infarction (MI) patients concerning the causes of MI. The present study also investigates the association between illness attributions and emotional adjustment, and the associations of relevant third variables with illness attributions. A sample of 37 wives were interviewed during the acute phase of the illness, three months and 10 years post-index MI. Attributions of the MI to multiple causes were common with an strong emphasis on psychosocial risk factors, but attributional content showed a low consistency over time. In retrospect, the wives believed they could have done more to prevent the MI, but self-blame was not a significant issue. However, they blamed the patient for not taking preventive actions. Attributions were not related to background and illness characteristics or denial in any strong and consistent manner. In a short-term perspective, attributing the MI to stress causes and husbands' personality was associated with worse emotional adjustment. Also, the use of magical explanations and blame of the patient were related to a poorer emotional outcome for the wife. The causal attributions made by the wives during the acute phase did not influence the emotional adjustment after 10 years.


FXR overexpression alters adipose tissue architecture in mice and limits its storage capacity leading to metabolic derangements.
The bile acid-activated nuclear receptor, FXR (NR1H4), has been implicated in the control of lipid and energy metabolism, but its role in fat tissue, where it is moderately expressed, is not understood. In view of the recent development of FXR-targeting therapeutics for treatment of human metabolic diseases, understanding the tissue-specific actions of FXR is essential. Transgenic mice expressing human FXR in adipose tissue (aP2-hFXR mice) at three to five times higher levels than endogenous Fxr, i.e., much lower than its expression in liver and intestine, have markedly enlarged adipocytes and show extensive extracellular matrix remodeling. Ageing and exposure to obesogenic conditions revealed a strongly limited capacity for adipose expansion and development of fibrosis in adipose tissues of aP2-hFXR transgenic mice. This was associated with impaired lipid storage capacity, leading to elevated plasma free fatty acids and ectopic fat deposition in liver and muscle as well as whole-body insulin resistance. These studies establish that adipose FXR is a determinant of adipose tissue architecture and contributes to whole-body lipid homeostasis.


Bicalutamide ('Casodex') 150 mg as adjuvant to radiotherapy in patients with localised or locally advanced prostate cancer: results from the randomised Early Prostate Cancer Programme.
The ongoing Early Prostate Cancer (EPC) programme is assessing bicalutamide ('Casodex') 150 mg, either alone or as adjuvant to treatment of curative intent, in patients with localised or locally advanced prostate cancer (n=8113). This paper presents an exploratory analysis of the subgroup of the EPC programme who received radiotherapy with curative intent (n=1370) in order to determine the efficacy (in terms of progression-free survival [PFS]) and tolerability of bicalutamide 150 mg in this setting. 1370 patients with T1-4, MO, any N prostate cancer received bicalutamide 150 mg or placebo adjuvant to radiotherapy of curative intent. This analysis was undertaken at median 5.3 years' follow-up. In patients with locally advanced disease (n=305), bicalutamide adjuvant to radiotherapy significantly increased PFS by 53% (event-time ratio 1.53; 95% confidence intervals [CI] 1.16, 2.02) compared with placebo and reduced the risk of disease progression by 42% (hazard ration [HR] 0.58; 95% CI 0.41, 0.84; P=0.00348). In these patients, objective progression was experienced by 33.5% of those randomised to bicalutamide versus 48.6% for those randomised to placebo. The between-group difference in patients with localised disease (n=1065) failed to reach statistical significance (HR 0.80; 95% CI 0.62, 1.03; P=0.088). The most common adverse events were breast pain (74.8%) and gynaecomastia (66.6%), which were mild to moderate in >90% of cases. Bicalutamide 150 mg/day given as adjuvant to radiotherapy significantly improved PFS in patients with locally advanced prostate cancer. For patients with localised disease, the results at this stage from the radiotherapy subgroup and the overall EPC programme suggest that adjuvant hormonal therapy is currently not appropriate. There were no unexpected tolerability findings.


Does the Use of Negative Pressure Wound Therapy and Postoperative Drains Impact the Development of Surgical Site Infections?: A PARITY Trial Secondary Analysis.
Surgical site infections (SSIs) represent a major complication following oncologic reconstructions. Our objectives were (1) to assess whether the use of postoperative drains and/or negative pressure wound therapy (NPWT) were associated with SSIs following lower-extremity oncologic reconstruction and (2) to identify factors associated with the duration of postoperative drains and with the duration of NPWT. This is a secondary analysis of the Prophylactic Antibiotic Regimens in Tumor Surgery (PARITY) trial, a multi-institution randomized controlled trial of lower-extremity oncologic reconstructions. Data were recorded regarding the use of drains alone, NPWT alone, or both NPWT and drains, including the total duration of each postoperatively. We analyzed postoperative drain duration and associations with tourniquet use, intraoperative thromboprophylaxis or antifibrinolytic use, incision length, resection length, and total operative time, through use of a linear regression model. A Cox proportional hazards model was used to evaluate the independent predictors of SSI. Overall, 604 patients were included and the incidence of SSI was 15.9%. Postoperative drains alone were used in 409 patients (67.7%), NPWT alone was used in 15 patients (2.5%), and both postoperative drains and NPWT were used in 68 patients (11.3%). The median (and interquartile range [IQR]) duration of drains and of NPWT was 3 days (IQR, 2 to 5 days) and 6 days (IQR, 4 to 8 days), respectively. The use of postoperative drains alone, NPWT alone, or both drains and NPWT was not associated with SSI (p = 0.14). Increased postoperative drain duration was associated with longer operative times and no intraoperative tourniquet use, as shown on linear regression analysis (p < 0.001 and p = 0.03, respectively). A postoperative drain duration of ≥14 days (hazard ratio [HR], 3.6; 95% confidence interval [CI], 1.3 to 9.6; p = 0.01) and an operative time of ≥8 hours (HR, 4.5; 95% CI, 1.7 to 11.9; p = 0.002) were independent predictors of SSI following lower-extremity oncologic reconstruction. A postoperative drain duration of ≥14 days and an operative time of ≥8 hours were independent predictors of SSI following lower-extremity oncologic reconstruction. Neither the use of postoperative drains nor the use of NPWT was a predictor of SSI. Future research is required to delineate the association of the combined use of postoperative drains and NPWT with SSI. Therapeutic Level II. See Instructions for Authors for a complete description of levels of evidence.


NEW BIOTECHNOLOGICAL TREATMENTS FOR LIPID DISORDERS.
Disorders of lipid and lipoprotein metabolism play a central role in the pathogenesis of atherosclerotic cardiovascular diseases (CVDs). Despite the widespread use of efficacious lipid-modifying therapies, the residual risk of CVD remains unacceptably high. The purpose of this manuscript is to review the application of new technologies in the treatment of lipid disorders. New therapies work mostly at the gene expression level and are, therefore, different from traditional small-molecule drugs that work mainly by inhibiting already synthesized proteins. We will briefly lay out the function of the gene products targeted by the new agents. Then, we will organize our review of new biotechnological treatments by the molecular approach, namely: monoclonal antibodies, antisense oligonucleotides, small-interfering RNAs, and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated 9 (Cas9)-based genome editing. The paper concludes with the description of the current clinical studies and the perspectives for the use of these agents. (REV INVEST CLIN. 2018;70:244-54).


Management of ventriculo-gallbladder shunt in the presence of gallstones.
Hydrocephalus is a prevalent health problem that is frequently encountered by paediatric neurosurgeons during infancy and childhood. We report a case of an 11-year-old boy with high cerebrospinal fluid protein hydrocephalus secondary to optic glioma that required a ventriculoperitoneal (VP) shunt. The patient had multiple failures of VP shunt and developed massive ascites. Alternatively, the hydrocephalus was treated by ventriculo-gallbladder (VG) shunt in the presence of sludge which was removed from the gallbladder before placement of the shunt. After VG shunt insertion, the patient expressed signs of infection with elevated liver profile, which emphasised the presence of gallstones. While the shunt was kept in its place without any complications, the gallstones were successfully removed by an endoscopic retrograde cholangiopancreatography. In conclusion, the presence of sludge is not a contraindication for VG shunt placement, and, if the VG shunt was complicated with gallstones, it could be treated without the need for cholecystectomy.


Pollen carryover, pollinator movement, and spatial context impact the delivery of pollination services in apple orchards.
Assessing the relative contributions of different pollinator taxa to pollination services is a central task in both basic eco-evolutionary research and applied conservation and agriculture. To that end, many studies have quantified single-visit pollen deposition and visitation frequency, which together determine a pollinator species' rate of conspecific pollen delivery. However, for plant species that require or benefit from outcrossing, pollination service quality further depends upon the ratio of outcross to self-pollen deposited, which is determined by two additional pollinator traits: pollen carryover and movement patterns among genetically compatible plant individuals. Here, we compare the pollination capacities of managed honey bees, native bumble bees, and native mining bees in apple-a varietally self-incompatible commercial crop-when pollen carryover and pollinator movement patterns are considered. We constructed simulation models of outcross pollen deposition parameterized using empirically measured single-visit pollen deposition, visitation frequency, and probabilities of intertree movement exhibited by each pollinator type, as well as pollen carryover patterns simulated based on parameters reported in the literature. In these models, we also explicitly specified the spatial relationships among cross-compatible trees based on field-realistic orchard layout schemes. We found that estimated pollination service delivery was considerably reduced for all pollinator types when pollen carryover and pollinator movement patterns were considered, as compared to when only single-visit pollen deposition and visitation frequency were considered. We also found that the performance of different pollinator types varied greatly across simulated orchard layout schemes and pollen carryover scenarios, including one instance where bumble and mining bees reversed their relative rankings. In all simulations, native bumble and mining bees outperformed managed honey bees in terms of both outcross pollen delivery per unit time and per flower visited, with disparities being greatest under scenarios of low pollen carryover. We demonstrate the degree to which pollination studies may reach inaccurate conclusions regarding pollination service delivery when pollen carryover and pollinator movement patterns are ignored. Our finding of the strong context dependence of pollination efficiency, even within a single plant-pollinator taxon pair, cautions that future studies in both basic and applied pollination biology should explicitly consider the ecological context in which pollination interactions take place.


Reducing the extinction risk of stochastic populations via nondemographic noise.
We consider nondemographic noise in the form of uncertainty in the reaction step size and reveal a dramatic effect this noise may have on the stability of self-regulating populations. Employing the reaction scheme mA→kA but allowing, e.g., the product number k to be a priori unknown and sampled from a given distribution, we show that such nondemographic noise can greatly reduce the population's extinction risk compared to the fixed k case. Our analysis is tested against numerical simulations, and by using empirical data of different species, we argue that certain distributions may be more evolutionary beneficial than others.


The spray-drying process is sufficient to inactivate infectious porcine epidemic diarrhea virus in plasma.
Porcine epidemic diarrhea virus (PEDV) is considered an emergent pathogen associated with high economic losses in many pig rearing areas. Recently it has been suggested that PEDV could be transmitted to naïve pig populations through inclusion of spray-dried porcine plasma (SDPP) into the nursery diet which led to a ban of SDPP in several areas in North America and Europe. To determine the effect of spray-drying on PEDV infectivity, 3-week-old pigs were intragastrically inoculated with (1) raw porcine plasma spiked with PEDV (RAW-PEDV-CONTROL), (2) porcine plasma spiked with PEDV and then spray dried (SD-PEDV-CONTROL), (3) raw plasma from PEDV infected pigs (RAW-SICK), (4) spray-dried plasma from PEDV infected pigs (SD-SICK), or (5) spray-dried plasma from PEDV negative pigs (SD-NEG-CONTROL). For the spray-drying process, a tabletop spray-dryer with industry-like settings for inlet and outlet temperatures was used. In the RAW-PEDV-CONTROL group, PEDV RNA was present in feces at day post infection (dpi) 3 and the pigs seroconverted by dpi 14. In contrast, PEDV RNA in feces was not detected in any of the pigs in the other groups including the SD-PEDV-CONTROL group and none of the pigs had seroconverted by termination of the project at dpi 28. This work provides direct evidence that the experimental spray-drying process used in this study was effective in inactivating infectious PEDV in the plasma. Additionally, plasma collected from PEDV infected pigs at peak disease did not contain infectious PEDV. These findings suggest that the risk for PEDV transmission through commercially produced SDPP is minimal.


Reversible switching of pRNA activity on the DNA packaging motor of bacteriophage phi29.
This paper reports a reversible switching of the biological activity of an RNA molecule, packaging RNA (pRNA), which is a central component of the DNA packaging motor of bacteriophage phi29. The switching mechanism contains two components: (1) inhibition of pRNA by a short antisense DNA (asDNA) that can bind to the 3' end of the pRNA and inactivate the packaging motor; and (2) reactivation of pRNA by isothermal removal of asDNA from pRNA through a strand displacement strategy. The switching process can be repeated for multiple cycles and has been demonstrated by gel electrophoresis and a virion assembly assay.


Proposal for a revised international standard series of patch tests.
Worldwide, there are 3 major standard series of patch tests, the European, North American and Japanese, together presenting 32 allergens, the differences between them being the result of regional variation in allergen distribution as well as differences in dermatological opinion. We propose a "minimal" international standard series of 20 allergens, together with an "extended" international standard series of 14 allergens.


Clinically Confirmed Stroke With Negative Diffusion-Weighted Imaging Magnetic Resonance Imaging: Longitudinal Study of Clinical Outcomes, Stroke Recurrence, and Systematic Review.
We sought to establish whether the presence (versus absence) of a lesion on magnetic resonance imaging (MRI) with diffusion weighting (DWI-MRI) at presentation with acute stroke is associated with worse clinical outcomes at 1 year. We recruited consecutive patients with a nondisabling ischemic stroke and performed DWI-MRI. Patients were followed up at 1 year to establish stroke recurrence (clinical or on MRI), cognitive impairment (Addenbrooke Cognitive Assessment Revised,<88) and modified Rankin Scale. A median of 4 days post stroke, one third (76/264; 29%) of patients did not have a DWI lesion (95% confidence interval, 23%-35%). There was no statistically significant difference between those with and without a DWI lesion with respect to age or vascular risk factors. Patients without a lesion were more likely to be women or have previous stroke. At 1 year, 11 of 76 (14%) patients with a DWI-negative index stroke had a clinical diagnosis of recurrent stroke or transient ischemic attack, 33% had cognitive impairment (Addenbrooke Cognitive Assessment Revised<88), and 40% still had modified Rankin Scale>1, no different from DWI-positive patients; DWI-positive patients were more likely to have a new lesion on MRI (14%), symptomatic or asymptomatic, than DWI-negative patients (2%; P=0.02). Our data were consistent with 6 other studies (total n=976), pooled proportion of DWI-negative patients was 21% (95% confidence interval, 12%-32%). Nearly one third of patients with nondisabling stroke do not have a relevant lesion on acute DWI-MRI. Patients with negative DWI-MRI had no better prognosis than patients with a lesion. DWI-negative stroke patients should receive secondary prevention.


A Large-Scale Analysis of the Relationship of Synonymous SNPs Changing MicroRNA Regulation with Functionality and Disease.
Historically, owing to not changing amino acid composition of protein sequences, synonymous mutations are commonly assumed to be neutral during evolution and therefore have no effect on the phenotype and disease. Here, based on observations from large-scale analysis of genomic data, we predicted the putative synonymous SNPs that could result in functional consequences and disease risk through changing the microRNA-mediated gene regulation. We found that nearly half of the synonymous SNPs could affect protein expression by changing microRNA regulation in human genome and these SNPs significantly prefer to be associated with human diseases and traits. The synonymous SNPs changing microRNA-mediated gene regulation tend to be more under recent positive selection, prefer to affect gene expression, and implicate in human disease. We conclude that the miRNA-mediated regulation changes could be a potential mechanism for the contributions of synonymous SNPs to protein functions and disease risks.


Simplified and Highly-reliable automated production of [18F]FSPG for clinical studies.
Background (S)-4-(3- 18 F-Fluoropropyl)-L-Glutamic Acid ([ 18 F]FSPG) is a positron emission tomography (PET) tracer that specifically targets the cystine/glutamate antiporter (xc-), which is frequently overexpressed in cancer and several neurological disorders. Pilot studies examining the dosimetry and biodistribution of ([ 18 F]FSPG in healthy volunteers and tumor detection in patients with non-small cell lung cancer, hepatocellular carcinoma, and brain tumors showed promising results. In particular, low background uptake in the brain, lung, liver, and bowel was observed that further leads to excellent imaging contrasts of [ 18 F]FSPG PET. However, reliable production-scale cGMP-compliant automated procedures for [ 18 F]FSPG production are still lacking to further increase the utility and clinical adoption of this radiotracer. Herein, we report the optimized automated approaches to produce [ 18 F]FSPG through two commercially available radiosynthesizers capable of supporting centralized and large-scale production for clinical use. Results Starting with activity levels of 60-85 GBq, the fully-automated process to produce [ 18 F]FSPG took less than 45 minutes with average radiochemical yields of 22.56 ± 0.97% and 30.82 ± 1.60% (non-decay corrected) using TRACERlab™ FXFN and FASTlab™, respectively. The radiochemical purities were > 95% and the formulated [ 18 F]FSPG solution was determined to be sterile and colorless with the pH of 6.5-7.5. No radiolysis of the product was observed up to 8 hours after final batch formulation. Conclusions In summary, cGMP-compliant radiosyntheses and quality control of [ 18 F]FSPG have been established on two commercially available synthesizers leveraging high activity concentration and radiochemical purity. While the clinical trials using [ 18 F]FSPG PET are currently underway, the automated approaches reported herein will accelerate the clinical adoption of this radiotracer and warrant centralized and large-scale production of [ 18 F]FSPG.


Endogenous adenosine modulates stimulation-induced depression at the frog neuromuscular junction.
1. Endogenous adenosine, which is produced by enzymatic degradation of ATP released from synaptic vesicles, has been shown to be a potent inhibitor of acetylcholine release from motor nerve terminals. It has been proposed that this auto-inhibition mechanism might contribute significantly to tetanic stimulation-induced depression. 2. Levels of facilitation and depression during a 20 Hz stimulus train differ greatly in different terminals, but are strongly and non-linearly correlated with the terminal's release characteristics (the amount of transmitter released per unit terminal length, or 'release efficacy'). There is a weaker, approximately linear, correlation between depression and release efficacy at 2 Hz stimulation. 3. The effects of both endogenous and exogenously applied adenosine are also highly variable for different nerve terminals. We have shown that much of this variability can be attributed to the release efficacy of each terminal in the case of endogenous effects, and to the size of the nerve terminal in the case of exogenously applied adenosine receptor agonists. 4. When nerve terminals are pooled according to their individual release characteristics, endogenous adenosine can be shown to contribute significantly to stimulation-induced depression of release primarily in terminals that release enough transmitter to generate significant levels of adenosine, but do not release so much transmitter that depletion of releasable quanta is severe.


Reconstruction of a case of thallium poisoning using LA-ICP-SFMS.
The unique capabilities of laser ablation in combination with inductively coupled plasma sector field mass spectrometry (LA-ICP-SFMS) were employed to reconstruct details of a homicide by thallium poisoning, which took place 38 years ago in Austria. Thallium was determined in several human bone samples after acid digestion in a microwave oven. The ICP-SFMS results showed that the thallium concentration in the victim's bones was in the range 1.07-2.63 microg g(-1), which is up to 170 times higher compared to concentrations found in bones of persons who have died due to natural causes. The results were in accordance with the values obtained by graphite furnace atomic absorption spectrometry (GF-AAS). Laser ablation ICP-SFMS was applied to assess the time interval between the victim's poisoning and death. Several line scans with a laser spot size of 50 mum were performed on a thumbnail of the poisoned person and on a reference thumbnail by laser ablation ICP-SFMS. Thallium peaks were detected on the nail of the victim at a distance of 2.5 mm from the younger edge of the nail.


A S-Layer Protein of Bacillus anthracis as a Building Block for Functional Protein Arrays by In Vitro Self-Assembly.
S-layer proteins create a cell-surface layer architecture in both bacteria and archaea. Because S-layer proteins self-assemble into a native-like S-layer crystalline structure in vitro, they are attractive building blocks in nanotechnology. Here, the potential use of the S-layer protein EA1 from Bacillus anthracis in constructing a functional nanostructure is investigated, and apply this nanostructure in a proof-of-principle study for serological diagnosis of anthrax. EA1 is genetically fused with methyl parathion hydrolase (MPH), to degrade methyl parathion and provide a label for signal amplification. EA1 not only serves as a nanocarrier, but also as a specific antigen to capture anthrax-specific antibodies. As results, purified EA1-MPH forms a single layer of crystalline nanostructure through self-assembly. Our chimeric nanocatalyst greatly improves enzymatic stability of MPH. When applied to the detection of anthrax-specific antibodies in serum samples, the detection of our EA1-MPH nanostructure is nearly 300 times more sensitive than that of the unassembled complex. Together, it is shown that it is possible to build a functional and highly sensitive nanosensor based on S-layer protein. In conclusion, our present study should serve as a model for the development of other multifunctional nanomaterials using S-layer proteins.


Incidence of thyroid diseases in Zhejiang Province, China, after 15 years of salt iodization.
Thyroid diseases(TD) can be induced by either deficient or excessive iodine intake. Universal Salt Iodization(USI) program has been implemented in China since 1995, to prevent iodine deficiency disorders (IDD). To evaluate the current conditions of TD and the role of USI, a multi-stage stratified random sampling scheme was used to perform a cross-sectional survey on the incidence of TD among participants in 6600 households in Zhejiang Province, a coastal area in China. Iodine nutrition status of the population was assessed by dietary iodine intake recall and urinary iodine concentration (UIC) of the participants, and TD were diagnosed by thyroid ultrasonography for 15122 participants and for 5873 participants by serum criteria for thyroid function(fT3, fT4, TSH, TRAb, TgAb, TPOAb; see Introduction for abbreviations). The median UIC of the surveyed population was 163μg iodine/L. From the participants 23.2% had UIC<100μg/L which is moderately iodine-deficient according to WHO classification. Diffuse goiter was present in 2.3% of the population and thyroid nodule in 20.9%. The incidence of hyperthyroidism, subclinical hyperthyroidism, hypothyroidism, subclinical hypothyroidism, Graves' disease and chronic lymphocytic thyroiditis was 0.5%, 0.6%, 0.6%, 7.8%, 0.2% and 0.3%, respectively. The proportion of several TD for participants with non-iodized salt intake was higher than that for participants with iodized salt intake.


New directions for migration policy in Europe.
There is a growing debate about the future direction of migration policy in Europe. After nearly 30 years of pursuing restrictive immigration and asylum policies, many European Union (EU) governments are beginning to re-assess their migration policies and to call for a new approach. For the first time in many years, several EU governments have begun to talk again about the benefits of labour migration and, even more significantly, have even begun to take action to recruit more migrants, especially skilled workers. This paper looks at the background to current calls for a new approach to migration in Europe and public reaction to these new initiatives. It first describes recent trends in migration in Europe and then briefly considers the demographic case for more migration. This is followed by a brief outline of some of the measures being considered by European governments to promote selective labour migration. The remainder of the paper is devoted to a discussion of some of the implications of this change in policy, focusing on two main issues: the likely consequences for sending countries, and the implications for the fight against the smuggling and trafficking of people.


Bioengineering approach to non-invasive measurement of body composition.
Measurement of body fat percentage is essential for medical care and research. The "gold standard" method for humans is underwater weighing, which is clearly inappropriate for infants, sick people and non-human animals. The corresponding criterion method for animals is comminution of the carcass followed by extraction of the fat with a volatile solvent such as ether. Our goal has been to develop a method for body composition (fat percentage) for use in animals and humans which is non-invasive and minimally intrusive, independent of variation in body conformation and fat distribution, and reasonable in cost. In one variant, our approach to this problem has been to move Archimedes' principle "on to dry land." The subject's volume is determined by measuring the differential buoyancy in comfortably breathable light (low density) and heavy atmospheres. In another, we use "structured light," in which a pattern of illumination is cast on the patient. The image is acquired using a video camera and the geometrical spatial coordinates of a large number of points on the surface of the subject are acquired. This permits the computation of the surface area and volume of the subject; which, combined with the weight, determines the fat percentage.


Clinical profile and antibiotics response in typhoid fever.
The objective of this study is to evaluate the clinical profile and drug response in typhoid fever. This is a retrospective analysis of paediatric patients suffering from typhoid fever who were admitted at Kathmandu Medical College Teaching Hospital, Sinamangal during the period of two years and nine months. Total numbers of 100 cases of typhoid were studied. Diagnosis of Typhoid fever was based on clinical features, Widal test and blood culture. The sensitivity pattern of drugs in blood culture was recorded. The mode of presentation, treatment history, laboratory investigations reports, antibiotics administered and response to therapy were recorded. Quinolone is still the highly sensitive drug and most widely used for Salmonella typhi. Because of the indiscriminate use of these drugs, resistant to ciprofloxacin has been quite high and the duration of the defeverscence period has also been prolonged. But Ofloxacin is still showed highly effective and widely used with good response.


Surgical management of benign gastric outlet obstruction in the elderly.
Surgical management of gastric outlet obstruction may associate with significant morbidity and mortality. Few studies have documented surgical outcomes in elderly patients. The aim of this study was to review recent operative results of benign gastric outlet obstruction in elderly patients compared with younger patients. Forty- seven consecutive patients from January 2000 through September 2008 were included. Preoperative, intraoperative data and early postoperative complications were analyzed. Fifteen operations were performed in elderly patients and 32 in younger patients. More patients in the elderly group were assigned as ASA class 3 (p=0.037), but Charlson comorbidity index was similar. Procedure types included Finney or Jaboulay pyloroplasty (n=26), antrectomy (n=13) and gastrojejunostomy (n=8). The mean postoperative hospital stay was 14.9 days. A modest correlation between the length of stay and the patient's age (p=0.044; r=0.294) was observed. There were two in hospital mortalities and four patients had complications. Surgery for benign gastric outlet obstruction is safe in the elderly population and is not associated with any increase in morbidity or mortality.


Enhanced SOCS3 in osteoarthiritis may limit both proliferation and inflammation.
Osteoarthritis (OA) is a degenerative joint disease that is characterized by localized inflammatory and secondary proliferative changes. Suppressor of cytokine signaling 3 (SOCS3) is elevated during OA development. We investigated the effects of this protein on human chondrocyte survival in OA and the inflammatory response together with the mechanisms of these effects. Small interfering RNA (siRNA) was used to knock down the expression of SOCS3 in interleukin(IL)-1β-induced primary human osteoarthritic chondrocytes. We found that siRNA-mediated SOCS3 knock-down in human osteoarthritic chondrocytes increased production of IL-1β-induced prostaglandin E2, cell growth, transcript level and nuclear translocation of cyclin D1. Silencing of SOCS3 resulted in altered expression of nuclear factor-kappa-B (NF-κB) and cyclooxygenase (COX2). Our findings indicate that enhanced SOCS3 could have contradictory influences on OA development. SOCS3 might protect damaged joints by its anti-inflammatory effect and by inhibition of over-augmented cartilage tissue repair, which could exhibit inhibitory properties for joint inflammation, abnormal chondrocyte clustering and osteophyte formation in OA. On the other hand, SOCS3 might reduce chondrocyte growth response, which would delay repair of subchondral cancellous bone damage in OA owing to its anti-proliferation effect. The anti-inflammation and growth inhibition effects exhibited by enhanced SOCS3 in OA appear to be related to its capacity to down-regulate expression levels of NF-κB and COX2.


Burden of excess mortality after implementation of the new kidney allocation system may be borne disproportionately by middle-aged recipients.
Under the new kidney allocation system (KAS), implemented in 2014, the distribution of the best quality donor kidney grafts shifted between age groups, but it is unclear whether this change translates to meaningful differences in post-transplant outcomes. We conducted a retrospective cohort study of 20,345 deceased donor kidney transplant recipients before and 4,605 recipients after implementation of the KAS using data from the United Network of Organ Sharing. Overall, two-year mortality was greater among recipients in the post-KAS era compared with the pre-KAS era (6.31% vs 5.91% respectively, [p = 0.01]), and two-year graft loss was not significantly different between eras (9.95% and 9.65%, respectively [p = 0.13]). In analysis stratified by age group (18-45, 46-55, 56-65, and ≥66 years), relative risk of mortality was 1.48 (95% confidence interval [CI] 1.09-1.98) among recipients 46-55 years old and 1.47 (95% CI 1.18-1.81) among recipients 56-65 years old. Relative risk of all-cause graft loss was 1.43 (95% CI 1.20-1.70) among recipients 56-65 years old. There were no significant differences in relative risk of mortality or graft loss associated with the KAS era among other age groups. After adjustment for recipient characteristics and characteristics of the changing donor pool, relative risk of two-year mortality and graft loss associated with the post-KAS era was attenuated for recipients aged 46-55 and 56-65 years, but remained statistically significant. In this early analysis after implementation of the KAS, there is suggestion that increased risk of mortality and graft loss may be disproportionately borne by middle-aged recipients, which is only partially accounted for by changes in recipient and donor characteristics. These findings signal a need to continue to monitor the effects of the KAS to ensure that allocation practices both maximize utility of the kidney graft pool and respect fairness between age groups.


Paediatric estimated average glucose in children with Type 1 diabetes.
Estimated average glucose has been used to transform HbA1c into a glucose measure that might better inform patients of their glycaemic control. The data set used to obtain the estimated average glucose equation was derived in adults with Type 1 and Type 2 diabetes, along with normal healthy control subjects, and requires testing in children. This was a cross-sectional study of 234 children and young people (106 male) with Type 1 diabetes aged 4.0-23.5 years who underwent continuous glucose monitoring over a 5-day period along with a measure of HbA1c . Regression analysis was used to determine estimated average glucose and agreement was assessed with the average glucose estimated from the Nathan equation: Nathan average glucose equation = 1.59 (HbA1c% ) - 2.59. Mean HbA1c was 76 mmol/mol (25.1) [9.1 (2.3)%] and mean continuous glucose monitoring tissue glucose was 10.4 (2.6) mmol/l. The relationship between continuous glucose monitoring tissue glucose and HbA1c was described by the paediatric equation: paediatric estimated average glucose = 0.49 (HbA1c %) + 5.95 (r = 0.45; P < 0.001). The mean paediatric estimated average glucose was 10.4 (1.1) mmol/l compared with that from the Nathan average glucose equation of 11.9 (3.7) mmol/l (P < 0.001). Overall, the paediatric estimated average glucose was 2.7 mmol/l lower than the Nathan estimated average glucose, with a 95% limit of agreement of ± 0.5 mmol/l. The agreement was very close with HbA1c values below 80 mmol/mol (9.5%). These data suggest that the Nathan estimated average glucose could be used in children and young people with Type 1 diabetes. Caution should still be exercised in the estimates derived for average glucose as the data set is skewed in both Nathan and paediatric average glucose estimates in opposite directions because of the differences in average HbA1c .


Exploring the Process of Neutrophil Transendothelial Migration Using Scanning Ion-Conductance Microscopy.
The dynamics of neutrophil transendothelial migration was investigated in a model of experimental septicopyemia. Scanning ion-conductance microscopy allowed us to determine changes in morphometric characteristics of endothelial cells during this process. In the presence of a pyogenic lesion simulated by Staphylococcus aureus, such migration was accompanied by both compensatory reactions and alteration of both neutrophils and endothelial cells. Neutrophils demonstrated crawling along the contact sites between endothelial cells, swarming phenomenon, as well as anergy and formation of neutrophil extracellular traps (NETs) as a normergic state. Neutrophil swarming was accompanied by an increase in the intercellular spaces between endothelial cells. Endothelial cells decreased the area of adhesion to the substrate, which was determined by a decrease in the cell projection area, and the cell membrane was smoothed. However, endothelial cell rigidity was paradoxically unchanged compared to the control. Over time, neutrophil migration led to a more significant alteration of endothelial cells: first, shallow perforations in the membrane were formed, which were repaired rather quickly, then stress fibrils were formed, and finally, endothelial cells died and multiple perforations were formed on their membrane.


A case of urethral duplication arising from the posterior urethra to the scrotum with urinary stone in a 6-year-old male.
Urethral duplication is a rare congenital anomaly. We report a 6-year-old male with type IIA2 (Y-type) using Effmann's classification. The accessory urethra, in which a urinary stone existed, arose from the posterior urethra to the scrotum. Because of recurrent urinary tract infection and urinary discharge from the accessory urethra, surgical removal of the accessory urethra through a scrotal incision was performed. At 7-month postoperative follow-up the patient was completely free from urinary incontinence and urinary tract infection.


Longitudinal relationships between self-compassion and depressive symptoms in midlife women.
Women in midlife face a range of biopsychosocial stressors that increase the risk of depression, with potential negative consequences in older adulthood. Lower self-compassion is known to predict subsequent depressive symptoms, but little is known about whether depressive symptoms predict subsequent levels of self-compassion. This study examined the longitudinal and reciprocal relationships between self-compassion and depressive symptoms over a 5-year period in midlife. This longitudinal study used data from 272 women aged 40-60 years at baseline. Cross-lagged panel analysis was used to investigate reciprocal relationships between depressive symptoms and self-compassion at baseline and 5-year follow-up. After controlling for autoregressive effects, self-compassion at baseline explained 4% of unique variance in depressive symptoms 5 years later. Baseline depressive symptoms also predicted subsequent levels of self-compassion, with 2% of unique variance explained. This is the first study to report a bidirectional longitudinal relationship between self-compassion and depressive symptoms. Midlife women may experience a negative cycle whereby low self-compassion and depressive symptoms reinforce each other through midlife and into older adulthood. Clinical trials examining self-compassion interventions in midlife are indicated.


Dose-dependent association of apolipoprotein E allele epsilon 4 with late-onset, sporadic Alzheimer's disease.
We examined the apolipoprotein E (apo E) genotypes in 47 patients with late-onset sporadic Alzheimer's disease (mean age at onset +/- standard deviation, 72.2 +/- 6.4 years), 8 with late-onset familial Alzheimer's disease (75.5 +/- 5.1 years), 18 with early-onset sporadic Alzheimer's disease (52.8 +/- 4.7 years), and 10 with early-onset familial Alzheimer's disease (52.0 +/- 6.8 years) in Japan and compared them with genotypes in control subjects. In late-onset sporadic Alzheimer's disease, apo E-epsilon 4 frequency increased significantly (epsilon 4 frequency: 0.34 vs 0.095 in controls, p < 0.0001), and the odds ratio, which represents the strength of association between Alzheimer's disease and apo E-epsilon 4, markedly increased with increasing dose of apo E-epsilon 4 gene (3 [95% confidence interval, 2-6] in one dose; 43 [95% confidence interval, 12-154] in two doses). This study also suggested that apo E-epsilon 4 is associated with both late-onset (epsilon 4: 0.31) and early-onset familial Alzheimer's disease (epsilon 4: 0.35). In contrast, we found no association between apo E-epsilon 4 and early-onset sporadic Alzheimer's disease (epsilon 4: 0.08). These results indicate that the risk of developing late-onset sporadic Alzheimer's disease is markedly dependent on the dose of apo E-epsilon 4, while apo E-epsilon 4 does not appear to be a major risk factor for early-onset sporadic Alzheimer's disease.


Percutaneous Transgastric Duodenal Stenting and Gastrostomy Repair Using a Vascular Closure Device: Proof of Concept in a Porcine Model.
Introduction. Gastroduodenal stenting is efficacious and safe in both benign and malignant foregut diseases. Transgastric duodenal stenting has been described and however requires a gastrostomy tube to remain in situ for 4 to 6 weeks post-procedure which can lead to complications. We present a technique for immediate gastric repair using a suture-mediated vascular closure device, without the need for a gastrostomy tube in porcine models. Methods. Percutaneous access into the stomach was achieved using fluoroscopy. Two or 3 Perclose Proglide devices were pre-deployed. The tract was dilated and a wire advanced into the distal duodenum. A 15.5 cm covered enteric stent was delivered through the gastrostomy, deployed and position confirmed. The gastrostomy was closed using Perclose Proglide sutures. Necropsy leak pressure measurement was performed to assess integrity of gastrostomy closure in the porcine models. Results. Two (n = 8) or 3 (n = 2) Perclose Proglide devices were deployed in ten porcine models, with 1 misfire (4.5%). Percutaneous transgastric access and stent delivery was successful in all porcine models. Mean leak pressure in the animals with adequately deployed devices was 219 mmHg (range 172 mmHg-270 mmHg). Conclusion. This study demonstrates percutaneous transgastric duodenal stenting with immediate gastric repair using suture-mediated vascular closure devices is a feasible procedure.


[The main variants of organic mental disorders comorbid with alcohol dependence].
To identify typical clinical and pathogenetic variants of organic mental disorders (OMD), comorbid with alcohol dependence. A study included 302 men who underwent forensic-psychiatric examination. Clinical/psychopathological, clinical/dynamic and statistical methods were used. RESULTS AND СONCLUSION: Significant disorders of social adaptation, commitment of serious unlawful acts, problems of diagnosis and forensic psychiatric assessment were typical of all comorbid disorders studied. We determined two main variants of this comorbid pathology: 1) primary OMD, complicated with alcohol dependence; 2) alcohol dependence, which caused secondary OMD. These variants differ not only in the etiopathogenesis, but in the clinical design and dynamic of disorders.


Cognitive and adaptive advantages of growth hormone treatment in children with Prader-Willi syndrome.
People with Prader-Willi syndrome (PWS) typically have mild to moderate intellectual deficits, compulsivity, hyperphagia, obesity, and growth hormone deficiencies. Growth hormone treatment (GHT) in PWS has well-established salutatory effects on linear growth and body composition, yet cognitive benefits of GHT, seen in other patient groups, have not been well studied in PWS. Study 1 included 96 children and youth with PWS aged 4-21 years who naturalistically varied in their exposures to GHT. Controlling for socioeconomic status, analyses compared cognitive and adaptive behavior test scores across age-matched treatment naïve versus growth hormone treated children. Study II assessed if age of treatment initiation or treatment duration was associated with subsequent cognition or adaptive behavior in 127, 4- to 21-year olds with PWS. Study III longitudinally examined cognitive and adaptive behavior in 168 participants who were either consistently on versus off GHT for up to 4-5 years. Compared to the treatment naïve group, children receiving GHT had significantly higher Verbal and Composite IQs, and adaptive communication and daily living skills. Children who began treatment before 12 months of age had higher Nonverbal and Composite IQs than children who began treatment between 1 and 5 years of age. Longitudinally, the groups differed in their intercepts, but not slopes, with each group showing stable IQ and adaptive behavior scores over time. Cognitive and adaptive advantages should be considered an ancillary benefit and additional justification for GHT in people with PWS. Future efforts need to target apparent socioeconomic inequities in accessing GHT in the PWS population.


PIRATE project: point-of-care, informatics-based randomised controlled trial for decreasing overuse of antibiotic therapy in Gram-negative bacteraemia.
Antibiotic overuse drives antibiotic resistance. The optimal duration of antibiotic therapy for Gram-negative bacteraemia (GNB), a common community and hospital-associated infection, remains unknown and unstudied via randomised controlled trials (RCTs). This investigator-initiated, multicentre, non-inferiority, informatics-based point-of-care RCT will randomly assign adult hospitalised patients receiving microbiologically efficacious antibiotic(s) for GNB to (1) 14 days of antibiotic therapy, (2) 7 days of therapy or (3) an individualised duration determined by clinical response and 75% reduction in peak C reactive protein (CRP) values. The randomisation will occur in equal proportions (1:1:1) on day 5 (±1) of efficacious antibiotic therapy as determined by antibiogram; patients, their physicians and study investigators will be blind to treatment duration allocation until the day of antibiotic discontinuation. Immunosuppressed patients and those with GNB due to complicated infections (endocarditis, osteomyelitis, etc) and/or non-fermenting bacilli (Acinetobacter spp, Burkholderia spp, Pseudomonas spp) Brucella spp, Fusobacterium spp or polymicrobial growth with Gram-positive organisms will be ineligible. The primary outcome is incidence of clinical failure at day 30; secondary outcomes include clinical failure, all-cause mortality and incidence of Clostridiumdifficile infection in the 90-day study period. An interim safety analysis will be performed after the first 150 patients have been followed for ≤30 days. Given a chosen margin of 10%, the required sample size to determine non-inferiority is roughly 500 patients. Analyses will be performed on both intention-to-treat and per-protocol populations. Ethics approval was obtained from the cantonal ethics committees of all three participating sites. Results of the main trial and each of the secondary endpoints will be submitted for publication in a peer-reviewed journal. This trial is registered at www.clinicaltrials.gov (NCT03101072; pre-results).


Effects of systemic lidocaine versus magnesium administration on postoperative functional recovery and chronic pain in patients undergoing breast cancer surgery: A prospective, randomized, double-blind, comparative clinical trial.
We aimed to compare the effects of intraoperative lidocaine and magnesium on postoperative functional recovery and chronic pain after mastectomy due to breast cancer. Systemic lidocaine and magnesium reduce pain hypersensitivity to surgical stimuli; however, their effects after mastectomy have not been evaluated clearly. In this prospective, double-blind, clinical trial, 126 female patients undergoing mastectomy were randomly assigned to lidocaine (L), magnesium (M), and control (C) groups. Lidocaine and magnesium were administered at 2 mg/kg and 20 mg/kg for 15 minutes immediately after induction, followed by infusions of 2 mg/kg/h and 20 mg/kg/h, respectively. The control group received the same volume of saline. Patient characteristics, perioperative parameters, and postoperative recovery profiles, including the Quality of Recovery 40 (QoR-40) survey, pain scales, length of hospital stay, and the short-form McGill pain questionnaire (SF-MPQ) at postoperative 1 month and 3 months were evaluated. The global QoR-40 scores on postoperative day 1 were significantly higher in group L than in group C (P = 0.003). Moreover, in sub-scores of the QoR-40 dimensions, emotional state and pain scores were significantly higher in group L than those in groups M and C (P = 0.027 and 0.023, respectively). At postoperative 3 months, SF-MPQ and SF-MPQ-sensitive scores were significantly lower in group L than in group C (P = 0.046 and 0.036, respectively). Intraoperative infusion of lidocaine improved the quality of recovery and attenuated the intensity of chronic pain in patients undergoing breast cancer surgery.


Splenic arterial interventions: anatomy, indications, technical considerations, and potential complications.
Splenic arterial interventions are increasingly performed to treat various clinical conditions, including abdominal trauma, hypersplenism, splenic arterial aneurysm, portal hypertension, and splenic neoplasm. When clinically appropriate, these procedures may provide an alternative to open surgery. They may help to salvage splenic function in patients with posttraumatic injuries or hypersplenism and to improve hematologic parameters in those who otherwise would be unable to undergo high-dose chemotherapy or immunosuppressive therapy. Splenic arterial interventions also may be performed to exclude splenic artery aneurysms from the parent vessel lumen and prevent aneurysm rupture; to reduce portal pressure and prevent sequelae in patients with portal hypertension; to treat splenic artery steal syndrome and improve liver perfusion in liver transplant recipients; and to administer targeted treatment to areas of neoplastic disease in the splenic parenchyma. As the use of splenic arterial interventions increases in interventional radiology practice, clinicians must be familiar with the splenic vascular anatomy, the indications and contraindications for performing interventional procedures, the technical considerations involved, and the potential use of other interventional procedures, such as radiofrequency ablation, in combination with splenic arterial interventions. Familiarity with the complications that can result from these interventional procedures, including abscess formation and pancreatitis, also is important.


Age-related evaluation of peripheral circulation of workers with vibration exposure.
This study was designed to clarify the age variation of the indices in the accelerated plethysmography (APG) for fingers and set the standard ageing curve. The indices of APG of workers with exposure to vibration were compared with those of the standard ageing curves. The subjects were 815 male workers, ranging in age from 18 to 66, without exposure to vibration and without history of hypertension, circulatory diseases or diabetes. Their indices of APG are used establishing the standard ageing curves. The workers occupationally exposed to hand-arm vibration were also the subjects to evaluate peripheral circulation of hand. The APG was measured on the index finger tip of his dominant side by an accelerated plethysmometer. The subjects were divided into age groups at intervals of 5 years. In all age groups from 18 to 64 years, each index of APG showed an approximately normal distribution in a logarithmic scale. The mode of the distribution shifted to a lower value with increasing age. The standard ageing curves were drawn by calculating 25th, 50th and 75th percentiles by a nonparametric method for each group. Each index of the standard ageing curves dropped with increasing age. The significant age effect was slightly marked below 30 years, but marked between 30 and 50 years. The indices of APG of workers with exposure to vibration were also compared with those of the standard ageing curve. The indices of -b/a and a/d of workers with exposure to vibration were deteriorated beyond physiological change. We established the standard ageing curves for APG for male men. The technique is good for the evaluation of peripheral circulation disorder of aged workers with occupational exposure to vibration.


In-office dispensing of oral oncolytics: a continuity of care and cost mitigation model for cancer patients.
The high cost of cancer therapies continues to lead to questions of affordability for the healthcare system and to patients. Ensuring patient access to oral cancer drugs presents a unique set of challenges due to the significant cost of these novel agents, healthcare/payer policies, and established distribution practices. The National Community Oncology Dispensing Association, Inc (NCODA) is a grassroots, nonprofit organization established by pharmacists who are directly involved at the community practice level in assisting patients with the acquisition of their oral cancer drugs. Community oncology practices that embrace the NCODA Quality Standards are able to provide exceptional patient care by providing direct access to oral cancer drugs through the in-practice dispensary. Patient continuity of care is ensured by allowing practice staff to manage all aspects of drug therapy-from initial dispense to completion of therapy-and in-practice dispensing allows for improved patient convenience, safety, and compliance. Practice staff in the dispensary area work directly with patients to address the insurance coverage limitations and financial toxicity of procuring these drugs. Medicare patients are not eligible to take advantage of patient assistance and/or co-pay programs that have been established by pharmaceutical companies. Foundations such as the Patient Access Network Foundation have been established to provide assistance to Medicare patients. This case report focuses on a new dispensary in a moderately sized oncology community practice that prescribes to the NCODA Quality Standards and outlines the processes developed to assist Medicare patients in accessing their oral cancer medications.


Acute Essex-Lopresti Syndrome: About a Case of Brachioradialis Tendon Transfer.
Background The classic treatment for acute Essex-Lopresti syndrome is closed reduction percutaneous pinning (CRPP) of the distal radioulnar joint (DRUJ). This work aimed to verify whether it was possible to add a transfer of the brachioradialis tendon to the pinning. Case Description The patient was a 39-year-old right-handed man, climbing instructor, who sustained the Mason II fracture and disjunction of the DRUJ. A transfer of the brachioradialis tendon severed from its muscle attach that was made through a bone tunnel passing through the radius and the neck of the ulna. The clinical and radiological result at the 6-month follow-up was satisfactory. Literature Review and Clinical Relevance Our results in a single case showed that the brachioradialis tendon transfer was useful in acute Essex-Lopresti syndrome.


A Novel Endoscopic Cerenkov Luminescence Imaging System for Intraoperative Surgical Navigation.
Cerenkov luminescence imaging is an emerging optical technique for imaging the distribution of radiopharmaceuticals in vivo. However, because of the light scattering effect, it cannot obtain optical information from deep internal organs. To overcome this challenge, we established a novel endoscopic Cerenkov luminescence imaging system that used a clinically approved laparoscope and an electron-multiplying charge-coupled device camera. We assessed the performance of the system through a series of in vitro and in vivo experiments. The results demonstrated superior superficial imaging resolution (0.1 mm), a large field of view (500 mm2 with 10 mm imaging distance), and superb imaging sensitivity (imaging 1 μCi) of our system. It captured the weak Cerenkov signal from internal organs successfully and was applied to intraoperative surgical navigation of tumor resection. It offered objective information of the tumor location and tumor residual during the surgical operation. This technique holds great potential for clinical translation.


Canadian psychiatrists' current attitudes, practices, and knowledge regarding fitness to drive in individuals with mental illness: a cross-Canada survey.
To assess current attitudes, practices, and knowledge of Canadian psychiatrists regarding fitness to drive in individuals with mental illness and to explore variations according to provincial legislation. We carried out a national cross-sectional survey, using a random sample of psychiatrists. We used a mail survey to collect data. In total, 248 psychiatrists participated; the response rate was 54.2% on traced subjects. The majority (64.1%) reported that they strongly agreed or agreed that addressing patients' fitness to drive is an important issue. However, only 18.0% of respondents were always aware of whether their patients were active drivers. One-fourth strongly agreed or agreed that they were confident in their ability to evaluate fitness to drive. In discretionary provinces, 29.3% of psychiatrists reported not knowing their provincial legislation, as did 14.6% of psychiatrists in mandatory provinces; of those responding, 54.0% from discretionary provinces and 2.8% from mandatory provinces gave incorrect answers. Psychiatrists' responses demonstrate a broad range of attitudes, practices, and knowledge. There appears to be a large gap between what is expected of psychiatrists and their readiness and self-perceived ability to make informed clinical decisions related to driving safety. There is a clear need for education and guidelines to assist psychiatrists in decision making about driving fitness.


Frequency of family dinner and adolescent body weight status: evidence from the national longitudinal survey of youth, 1997.
To explore associations between overweight status and the frequency of family dinners (FFD) for adolescents and how those associations differ across race and ethnicity. A sample of 5014 respondents between 12 and 15 years of age from the 1997 wave of the National Longitudinal Survey of Youth 1997 (NLSY97) was used. BMI was calculated using self-reported height and weight; 13.3% of respondents qualified as overweight, 16.4% qualified as at-risk-of-overweight, and 1.9% qualified as underweight. The remainder were normal weight. FFD was defined as the number of times respondents had dinner with their families in a typical week in the past year. Multinomial logistic regression models were estimated separately for non-Hispanic whites vs. blacks and Hispanics for odds of belonging to the other weight categories compared with normal weight. A supplementary longitudinal analysis estimated the odds of change in overweight status between 1997 and 2000. In 1997, the FFD distribution was as follows: 0, 8.3%; 1 or 2, 7.3%; 3 or 4, 13.4%; 5 or 6, 28.1%; 7, 42%. For whites, higher FFD was associated with reduced odds of being overweight in 1997, reduced odds of becoming overweight, and increased odds of ceasing to be overweight by 2000. No such associations were found for blacks and Hispanics. Reasons for racial and ethnic differences in the relationship between FFD and overweight may include differences in the types and portions of food consumed at family meals. More research is needed to verify this.


A quantitative histological analysis of the effects of age and sex on human lingual epithelium.
After exclusion criteria had eliminated changes of pathological origin, tongues were obtained from 86 necropsies evenly divided by sex and age from 16-98 years. Epithelium was examined from two precisely defined sites on the lateral and dorsal lingual surfaces. Morphometric techniques were employed to determine various epithelial parameters. The epithelium was thicker in males but rates of ageing change between the sexes were similar. Generally, age changes were similar at each site: the mean epithelial thickness underwent a 30% reduction over the age range studied; the progenitor cell layer remained of constant thickness but its nuclear/cytoplasm ratio reduced significantly; significant reductions also occurred in the rete surface area of the lateral epithelium and in the papillary surface area of the dorsal epithelium. Although some of the parameters showed steeper rates of change in the younger half of the age range, the overall pattern of ageing in the lingual epithelium suggested a continuous trend towards atrophy and simplification of structure occurring evenly throughout the entire adult life span.


Granuloma associated with Trichosporon asahii infection in the lung: Unusual pathological findings and PCR detection of Trichosporon DNA.
We describe the first case of a Trichosporon asahii infection of the lung characterized by an epithelioid granuloma with multinucleated giant cells. The patient died of respiratory failure due to lung fibrosis and lung cancer. Autopsy showed primary lung cancer with metastasis, lung fibrosis, and a focal infectious lesion containing the filamentous and yeast forms of a fungus. From its shape the fungus was judged to be Trichosporon, but unusual pathological findings revealed an epithelioid granuloma with multinucleated giant cells surrounding the hyphae and yeast cells. Immunohistochemical staining was performed using the specific Trichosporon spp. monoclonal antibodies and DNA identificationn was done by the PCR. The results showed this case to be a localized Trichosporon asahii infection. Hypersensitivity pneumonitis (HP) due to Trichosporon spp. with its pathological finding of inflammation with granuloma is common, but infections with granulomatous inflammation are extremely rare. This case suggests that Trichosporon infection may develop into granuloma with multinucleated giant cells.


Outbreak of Zika virus infection in Singapore: an epidemiological, entomological, virological, and clinical analysis.
An outbreak of Zika virus infection was detected in Singapore in August, 2016. We report the first comprehensive analysis of a national response to an outbreak of Zika virus infection in Asia. In the first phase of the outbreak, patients with suspected Zika virus infection were isolated in two national referral hospitals until their serum tested negative for the virus. Enhanced vector control and community engagement measures were deployed in disease clusters, including stepped-up mosquito larvicide and adulticide use, community participation in source reduction (destruction of mosquito breeding sites), and work with the local media to promote awareness of the outbreak. Clinical and epidemiological data were collected from patients with confirmed Zika virus infection during the first phase. In the second phase, admission into hospitals for isolation was stopped but vector control efforts continued. Mosquitoes were captured from areas with Zika disease clusters to assess which species were present, their breeding numbers, and to test for Zika virus. Mosquito virus strains were compared with human strains through phylogenetic analysis after full genome sequencing. Reproductive numbers and inferred dates of strain diversification were estimated through Bayesian analyses. From Aug 27 to Nov 30, 2016, 455 cases of Zika virus infection were confirmed in Singapore. Of 163 patients with confirmed Zika virus infection who presented to national referral hospitals during the first phase of the outbreak, Zika virus was detected in the blood samples of 97 (60%) patients and the urine samples of 157 (96%) patients. There were 15 disease clusters, 12 of which had high Aedes aegypti breeding percentages. Captured mosquitoes were pooled into 517 pools for Zika virus screening; nine abdomen pools (2%) were positive for Zika virus, of which seven head and thorax pools were Zika-virus positive. In the phylogenetic analysis, all mosquito sequences clustered within the outbreak lineage. The lineage showed little diversity and was distinct from other Asian lineages. The estimated most recent common ancestor of the outbreak lineage was from May, 2016. With the deployment of vector control and community engagement measures, the estimated reproductive number fell from 3·62 (95% CI 3·48-3·77) for July 31 to Sept 1, 2016, to 1·22 (95% CI 1·19-1·24) 4 weeks later (Sept 1 to Nov 24, 2016). The outbreak shows the ease with which Zika virus can be introduced and spread despite good baseline vector control. Disease surveillance, enhanced vector control, and community awareness and engagement helped to quickly curb further spread of the virus. These intensive measures might be useful for other countries facing the same threat. National Medical Research Council Singapore, Centre for Infectious Disease Epidemiology and Research, and A*STAR Biomedical Research Council.


One set of pliers for more tasks in installation work: the effects on (dis)comfort and productivity.
In installation work, the physical workload is high. Awkward postures, heavy lifting and repetitive movements are often seen. To improve aspects of the work situation, frequently used pliers were redesigned to make them suitable for more cutting tasks. In this study these multitask pliers are evaluated in comparison to the originally used pliers in a field study and a laboratory study. For the field study 26 subjects participated divided into two groups according to their type of work. Ten subjects participated in the laboratory study. The multitask plier appeared to result in more comfort during working, more relaxed working and more satisfaction. No differences in productivity were found. In conclusion, the multitask pliers can replace the originally used pliers and are suitable for more tasks than the original pliers. The installation workers have to carry less pliers by using the multitask pliers.


The CDX2 transcription factor regulates furin expression during intestinal epithelial cell differentiation.
CDX2, a member of the caudal family of transcription factors, is involved in enterocyte lineage specification. CDX2 activates many intestine-specific genes, such as sucrase-isomaltase and lactase-phlorizin hydrolase (LPH), and adhesion proteins, namely, LI-cadherin and claudin-2. In this study, we show that the proprotein convertase furin, involved in proteolytic maturation of proprotein substrates including LPH and cell surface proteins, is a CDX2 target. Indeed, expression of the rat furin homolog was induced 1.5-fold, as determined by microarray experiments that compared control with CDX2-expressing intestinal epithelial cells (IEC-6). As determined by transient transfection assays in Caco-2/15 cells, the furin P1 promoter 1.3-kb fragment between SacI and NheI was essential for CDX2 transcriptional activation. Electrophoretic mobility shift/supershift assays followed by site-specific mutagenesis and chromatin immunoprecipitation identified the CDX DNA-binding site (CBS)2 sequence from nt -1827 to -1821 as the major CBS involved in furin P1 promoter activation. Increased furin mRNA and protein expression correlated with both CDX2 expression and intestinal epithelial cell differentiation. In addition, furin mRNAs were detected predominantly in differentiated epithelial cells of the villus, as determined by in situ hybridization. Treatment of Caco-2/15 cells with a furin inhibitor led to inhibition of LPH activity. Morphological differentiation of enterocyte-like features in Caco-2/15 such as epithelial cell polarity and brush-border formation were strongly attenuated by furin inhibition. These results suggest that CDX2 regulates furin expression in intestinal epithelial cells. Furin may be important in modulating the maturation and/or activation of key factors involved in enterocyte differentiation.


Investigating the structural and functional consequences of circular permutation on lipase B from Candida antarctica.
The engineering of lipase B from Candida antarctica (CALB) by circular permutation has yielded over sixty hydrolase variants, and several show significantly improved catalytic performance. Here we report a detailed characterization of ten selected enzyme variants by kinetic and spectroscopic methods to further elucidate the impact of circular permutation on the structure and function of CALB. Our experiments identify lipase variants with up to 175-fold enhanced k(cat)/K(M) values over wild-type. In addition, circular permutation does not change the enzymes' enantiopreference and preserves or even improves their enantioselectivity compared to that of the wild-type enzyme. Finally, our spectroscopic analyses suggest that the structural effects of circular permutation on CALB are mostly local, concentrating on regions near the native and new protein termini. The observed changes in secondary structure and protein thermostability vary among enzyme variants but directly correlate with the locations of the new termini, a first step towards a predictive framework.


Understanding the GERD.
Gastroesophageal reflux disease (GERD) is the most common gastrointestinal disorder in the United States. Without proper treatment, patients may be at risk for long-term complications including Barrett's esophagus and adenocarcinoma of the esophagus. Because reflux is a common complaint, clinicians need to know how to rule out causes other than GERD and how to treat patients suspected of having GERD. This article discusses how to diagnose GERD, the medical and surgical treatment options, and how to evaluate patients who are at risk for long-term complications.


Fifteen Years of Experience from a Medical School' Clinical Skills Laboratory.
Medical Education has evolved being defined as a continuum of the medical training from the pre- to the post-graduate period and through a lifetime of medical practice that is mirrored in the competencies framework that several international institutions have established. This creates a challenge to educational institutions (pre- and post-graduate) that traditionally take separate pathways. The present report is a description of the efforts carried by the School of Medicine of the University of Minho to implement a strategy of teaching/learning methods that follows modern standards towards constructive alignment of the medical curriculum, from the pre- to the post-graduate. The faculty members responsible for these activities make a narrative self-report of the activities developed and where available quantitative data from participant surveys are presented. In order to achieve a constructive alignment of educational/assessment strategies several steps were taken that focused on the revision of the learning goals, teaching methodologies and assessment strategies. This implicated the investment in the training/certification of faculty, acquisition of simulation tools and a dedicated infrastructure. Alumni were a fundamental cornerstone as voluntary faculty contributing to pre-graduate training, but also training their own scholar skills. Overall, courses are rate with a high rate of satisfaction among course participants. Setting up the present teaching/learning environment of the School of Medicine of the University of Minho required a collective effort of the faculty, as well a progressive investment in both acquisition of equipment and training of staff. These human and material efforts, however, lead to an excellent return in learning outcomes. The main conclusion is that the constructive alignment of educational and assessment strategies towards the medical education continuum needs reflective thinking on the learners' needs. The secondary gain of these initiatives is to provide opportunities for junior doctors to practice teaching.


A Low-Cost Pulse Generator for Exacerbating Muscle Fiber Detachment Phenotypes in Zebrafish.
Muscle fiber detachment from myoseptal boundaries is a common finding in zebrafish models of muscular dystrophies. In some instances, there is a weakening of the interaction between muscle fiber and myosepta, which is yet to manifest as a fiber detachment phenotype. Therefore, to push the fiber detachment of muscle, mutant fish but not their wild-type siblings, beyond their binding threshold, a series of small electrical pulses can be applied to the larvae to create a maximal force contraction and ultimately fiber detachment. To do this, we built a digital pulse generator which delivers four 8 ms 30 V pulses in quick succession, and it has the advantage over older analog approaches to pulse generation because it improves accuracy and is appreciably less expensive. Our pulse generator significantly increases fiber detachment in the laminin-α2 deficient, congenital muscular dystrophy type 1a (MDC1a) model lama2-/- fish when compared with controls.


Zein as an Effective Carrier for Hesperidin Delivery Systems with Improved Prebiotic Potential.
Hesperidin is a polyphenol derived from citrus fruits that has a broad potential for biological activity and the ability to positively modify the intestinal microbiome. However, its activity is limited by its low solubility and, thus, its bioavailability-this research aimed to develop a zein-based hesperidin system with increased solubility and a sustained release profile. The study used triple systems enriched with solubilizers to maximize solubility. The best system was the triple system hesperidin-zein-Hpβ-CD, for which the solubility improved by more than six times. A significant improvement in the antioxidant activity and the ability to inhibit α-glucosidase was also demonstrated, due to an improved solubility. A release profile analysis was performed in the subsequent part of the experiments, confirming the sustained release profile of hesperidin, while improving the solubility. Moreover, the ability of selected probiotic bacteria to metabolize hesperidin and the effect of this flavonoid compound on their growth were investigated.


[Symptom-centered group therapy for phobias--the problem-oriented workshop in psychotherapy].
Short term psychotherapy is increasingly recognized as a socioeconomic necessity. Problem-oriented treatments, particularly if applied in problem-homogenous groups, are one means of economizing and sometimes even improving psychotherapy. They also require a re-evaluation of the nature of the patient-therapist dyad. The "task-oriented small group" as a basic field of human interaction has been studied intensively in experimental social psychology. Nevertheless, group psychotherapy and behavior therapy have largely neglected group-specific aspects of such variables as cohesion, task-orientation, modelling, mutual self-help, risk-taking, motivation and insight. This paper outlines some recent changes in this respect, and discusses in particular the "symptomoriented peer workshop" as a treatment milieu. The general therapeutic potential of the peer workshop is briefly assessed with regard to the application of the above mentioned group variables in peer self-help groups, rehabilitation programs for the chronically hospitalized, symptom or depth-treatment in short-term psychotherapy, and in re-education programs for problem-children. The main concern is with its specific application to the treatment of phobias. Phobias are comparatively easy to define, yet are often connected with other personality problems. This has made them a model-problem in the arguments about symptom-substitution between "psycho"- and "behavior" -therapists, and it has stimulated discussion as to how behavior-focused treatments affect cognition and emotion, and to what extend depth and insight-oriented therapies change behavior. The interaction between treated and non-treated problems, and the behavioral, cognitive and emotional effects of differential treatment ingredients are discussed with special reference to the first controlled study in behavior therapy, that applied behavior-focussed confrontation with reality (flooding in vivo) under the sociodynamics of the small gsoup. Problem-focussed retraining under real-life conditions (reality testing) as group-therapeutic fieldwork can be a means of treatment in its own right may open up the patient for subsequent attempts to tackle more complex problems, or may just be an adjuvant in a multi-level approach. This certainly applies for phobias and can probably be generalized to other problems.


Investigation of serum proteome alterations in human glioblastoma multiforme.
Glioblastoma multiforme (GBM) or grade IV astrocytoma is the most common and lethal adult malignant brain tumor. The present study was conducted to investigate the alterations in the serum proteome in GBM patients compared to healthy controls. Comparative proteomic analysis was performed employing classical 2DE and 2D-DIGE combined with MALDI TOF/TOF MS and results were further validated through Western blotting and immunoturbidimetric assay. Comparison of the serum proteome of GBM and healthy subjects revealed 55 differentially expressed and statistically significant (p <0.05) protein spots. Among the identified proteins, haptoglobin, plasminogen precursor, apolipoprotein A-1 and M, and transthyretin are very significant due to their functional consequences in glioma tumor growth and migration, and could further be studied as glioma biomarkers and grade-specific protein signatures. Analysis of the lipoprotein pattern indicated elevated serum levels of cholesterol, triacylglycerol, and low-density lipoproteins in GBM patients. Functional pathway analysis was performed using multiple software including ingenuity pathway analysis (IPA), protein analysis through evolutionary relationships (PANTHER), database for annotation, visualization and integrated discovery (DAVID), and GeneSpring to investigate the biological context of the identified proteins, which revealed the association of candidate proteins in a few essential physiological pathways such as intrinsic prothrombin activation pathway, plasminogen activating cascade, coagulation system, glioma invasiveness signaling, and PI3K signaling in B lymphocytes. A subset of the differentially expressed proteins was applied to build statistical sample class prediction models for discrimination of GBM patients and healthy controls employing partial least squares discriminant analysis (PLS-DA) and other machine learning methods such as support vector machine (SVM), Decision Tree and Naïve Bayes, and excellent discrimination between GBM and control groups was accomplished.


[Cystic lymphangioma of the pancreas. Apropos of a case].
A new case of pancreatic cystic lymphangioma is reported in a young woman. The authors discuss its clinic and histologic features and emphasize the complementary findings of ultrasonography and CT scan which are similar to the other sites of cystic lymphangioma in adults.