Diverse type VI secretion phospholipases are functionally plastic antibacterial effectors.
Membranes allow the compartmentalization of biochemical processes and are therefore fundamental to life. The conservation of the cellular membrane, combined with its accessibility to secreted proteins, has made it a common target of factors mediating antagonistic interactions between diverse organisms. Here we report the discovery of a diverse superfamily of bacterial phospholipase enzymes. Within this superfamily, we defined enzymes with phospholipase A1 and A2 activity, which are common in host-cell-targeting bacterial toxins and the venoms of certain insects and reptiles. However, we find that the fundamental role of the superfamily is to mediate antagonistic bacterial interactions as effectors of the type VI secretion system (T6SS) translocation apparatus; accordingly, we name these proteins type VI lipase effectors. Our analyses indicate that PldA of Pseudomonas aeruginosa, a eukaryotic-like phospholipase D, is a member of the type VI lipase effector superfamily and the founding substrate of the haemolysin co-regulated protein secretion island II T6SS (H2-T6SS). Although previous studies have specifically implicated PldA and the H2-T6SS in pathogenesis, we uncovered a specific role for the effector and its secretory machinery in intra- and interspecies bacterial interactions. Furthermore, we find that this effector achieves its antibacterial activity by degrading phosphatidylethanolamine, the major component of bacterial membranes. The surprising finding that virulence-associated phospholipases can serve as specific antibacterial effectors suggests that interbacterial interactions are a relevant factor driving the continuing evolution of pathogenesis.


Persistence and generalization of adaptive changes in auditory localization behavior following unilateral conductive hearing loss.
Sound localization relies on the neural processing of binaural and monaural spatial cues generated by the physical properties of the head and body. Hearing loss in one ear compromises binaural computations, impairing the ability to localize sounds in the horizontal plane. With appropriate training, adult individuals can adapt to this binaural imbalance and largely recover their localization accuracy. However, it remains unclear how long this learning is retained or whether it generalizes to other stimuli. We trained ferrets to localize broadband noise bursts in quiet conditions and measured their initial head orienting responses and approach-to-target behavior. To evaluate the persistence of auditory spatial learning, we tested the sound localization performance of the animals over repeated periods of monaural earplugging that were interleaved with short or long periods of normal binaural hearing. To explore learning generalization to other stimulus types, we measured the localization accuracy before and after adaptation using different bandwidth stimuli presented against constant or amplitude-modulated background noise. Retention of learning resulted in a smaller initial deficit when the same ear was occluded on subsequent occasions. Each time, the animals' performance recovered with training to near pre-plug levels of localization accuracy. By contrast, switching the earplug to the contralateral ear resulted in less adaptation, indicating that the capacity to learn a new strategy for localizing sound is more limited if the animals have previously adapted to conductive hearing loss in the opposite ear. Moreover, the degree of adaptation to the training stimulus for individual animals was significantly correlated with the extent to which learning extended to untrained octave band target sounds presented in silence and to broadband targets presented in background noise, suggesting that adaptation and generalization go hand in hand. Together, these findings provide further evidence for plasticity in the weighting of monaural and binaural cues during adaptation to unilateral conductive hearing loss, and show that the training-dependent recovery in spatial hearing can generalize to more naturalistic listening conditions, so long as the target sounds provide sufficient spatial information.


The validity of using an electrocutaneous device for pain assessment in patients with cervical radiculopathy.
The purpose of this study was to evaluate the validity and preference for assessing pain magnitude with electrocutaneous testing (ECT) compared to the visual analogue scale (VAS) and Borg CR10 scale in men and women with cervical radiculopathy of varying sensory phenotypes. An additional purpose was to investigate ECT sensory and pain thresholds in men and women with cervical radiculopathy of varying sensory phenotypes. This is a cross-sectional study of 34 patients with cervical radiculopathy. Scatterplots and linear regression were used to investigate bivariate relationships between ECT, VAS and Borg CR10 methods of pain magnitude measurement as well as ECT sensory and pain thresholds. The use of the ECT pain magnitude matching paradigm for patients with cervical radiculopathy with normal sensory phenotype shows good linear association with arm pain VAS (R(2) = 0.39), neck pain VAS (R(2) = 0.38), arm pain Borg CR10 scale (R(2) = 0.50) and neck pain Borg CR10 scale (R(2) = 0.49) suggesting acceptable validity of the procedure. For patients with hypoesthesia and hyperesthesia sensory phenotypes, the ECT pain magnitude matching paradigm does not show adequate linear association with rating scale methods rendering the validity of the procedure as doubtful. ECT for sensory and pain threshold investigation, however, provides a method to objectively assess global sensory function in conjunction with sensory receptor specific bedside examination measures.


Landscape context and the biophysical response of rivers to dam removal in the United States.
Dams have been a fundamental part of the U.S. national agenda over the past two hundred years. Recently, however, dam removal has emerged as a strategy for addressing aging, obsolete infrastructure and more than 1,100 dams have been removed since the 1970s. However, only 130 of these removals had any ecological or geomorphic assessments, and fewer than half of those included before- and after-removal (BAR) studies. In addition, this growing, but limited collection of dam-removal studies is limited to distinct landscape settings. We conducted a meta-analysis to compare the landscape context of existing and removed dams and assessed the biophysical responses to dam removal for 63 BAR studies. The highest concentration of removed dams was in the Northeast and Upper Midwest, and most have been removed from 3rd and 4th order streams, in low-elevation (< 500 m) and low-slope (< 5%) watersheds that have small to moderate upstream watershed areas (10-1000 km2) with a low risk of habitat degradation. Many of the BAR-studied removals also have these characteristics, suggesting that our understanding of responses to dam removals is based on a limited range of landscape settings, which limits predictive capacity in other environmental settings. Biophysical responses to dam removal varied by landscape cluster, indicating that landscape features are likely to affect biophysical responses to dam removal. However, biophysical data were not equally distributed across variables or clusters, making it difficult to determine which landscape features have the strongest effect on dam-removal response. To address the inconsistencies across dam-removal studies, we provide suggestions for prioritizing and standardizing data collection associated with dam removal activities.


Low-dose secnidazole in the treatment of bacterial vaginosis.
To determine whether bacterial vaginosis could be cured with a single 1-g oral dose of secnidazole. A total of 80 women were recruited at the outpatient gynecologic clinic of Manuel Noriega Hospital, Maracaibo, Venezuela. Diagnosis and patient enrollment were based on the Amsel criteria. The participants were randomized to 2 groups. In group 1 (n=44) participants received a single 1-g oral dose and in group 2 (n=32) participants received a single 2-g oral dose of secnidazole. Clinical cure was defined as the absence of the characteristic symptoms (a bad odor and a grossly abnormal discharge) and at least 2 of the following: vaginal pH less than 4.5, no fish odor on addition of KOH, and no Gardnerella vaginalis or clue cells on wet-mount examination. Cytologic cure was defined as an absence of G. vaginalis on a Papanicolaou (Pap) smear. Clinical cure was experienced by 95.5% of the women who received the 1-g oral dose and by 97.4% of the women who received the 2-g oral dose of secnidazole. There was no significant difference between the groups in the clinical resolution of bacterial vaginosis. Following treatment, results were negative for G. vaginalis in 94.7% of the women. In group 1, 41 women (93.2%), and in group 2, 31 women (96.9%) had cytologic cure. The Pap smear revealed G. vaginalis in 3 of the women in group 1 and 1 of the women in group 2 (P=0.47). Twenty-seven women (35.5%) reported mild side effects. More women had adverse effects in group 1 (n=16) than in group 2 (n=11) but this difference was not statistically significant. This clinical study showed that a single 1-g oral dose of secnidazole is effective to cure bacterial vaginosis associated with G. vaginalis.


The Centers for Disease Control and Prevention's Healthy Days Measures - population tracking of perceived physical and mental health over time.
To promote the health and quality of life of United States residents, the U.S. Department of Health and Human Services' Centers for Disease Control and Prevention (CDC) - with 54 state and territorial health agencies - has supported population surveillance of health-related quality of life (HRQOL). HRQOL was defined as "perceived physical and mental health over time." Commonly-used measures of health status and activity limitation were identified and a set of "Healthy Days" HRQOL measures was developed and validated. A core set of these measures (the CDC HRQOL-4) asks about self-rated general health and the number of recent days when a person was physically unhealthy, mentally unhealthy, or limited in usual activities. A summary measure combines physically and mentally unhealthy days. From 1993 to 2001, more than 1.2 million adults responded to the CDC HRQOL-4 in each state-based Behavioral Risk Factor Surveillance System (BRFSS) telephone interview. More than one fifth of all BRFSS respondents also responded to a set of related questions - including five items that assess the presence, main cause and duration of a current activity limitation, and the need for activity-related personal and routine care; as well as five items that ask about recent days of pain, depression, anxiety, sleeplessness, and vitality. The Healthy Days surveillance data are particularly useful for finding unmet health needs, identifying disparities among demographic and socioeconomic subpopulations, characterizing the symptom burden of disabilities and chronic diseases, and tracking population patterns and trends. The full set of 14 Healthy Days Measures (the CDC HRQOL-14) has shown good measurement properties in several populations, languages, and settings. The brief standard CDC HRQOL-4 is now often used in surveys, surveillance systems, prevention research, and population health report cards.


Meta-analysis comparing bivalirudin versus heparin monotherapy on ischemic and bleeding outcomes after percutaneous coronary intervention.
With femoral access, bivalirudin decreases risks of major bleeding after percutaneous coronary intervention (PCI) and provides better net clinical benefit compared to unfractionated heparin (UFH) plus planned glycoprotein IIb/IIIa inhibitors. Whether this benefit exists compared to UFH monotherapy is less clear. We performed a systematic review and meta-analysis to compare outcomes in patients undergoing transfemoral PCI with UFH or bivalirudin. Randomized trials (n = 3) and observational studies (n = 13) comparing bivalirudin to UFH monotherapy were reviewed. Primary outcomes were 30-day rates of major adverse cardiovascular events (MACEs) including death, myocardial infarction (MI), urgent revascularization, as well as all-cause mortality, MI, major bleeding, and blood transfusion. We collected data from 16 studies involving 32,492 patients undergoing PCI. Most observational studies were performed in the United States, whereas all randomized trials were done in Europe. Compared to UFH monotherapy, bivalirudin was associated with similar risk of MACEs (odds ratios [OR] 0.92, 95% confidence interval [CI] 0.75 to 1.12), a substantial 45% relative decrease in major bleeding (OR 0.55, 95% CI 0.43 to 0.72), and a trend in the decrease of transfusion (OR 0.87, 95% CI 0.70 to 1.08). A decrease in mortality was seen in observational studies (OR 0.62, 95% CI 0.45 to 0.85) but remained inconclusive in randomized trials (OR 0.63, 95% CI 0.20 to 2.01). MI rate was similar with the 2 anticoagulants. In conclusion, in patients undergoing transfemoral PCI, the benefit of bivalirudin over UFH monotherapy is driven by a significant decrease in major bleeding with similar rates of MACE. As PCI practice moves toward other bleeding-avoidance strategies such as the radial approach, future studies should focus on the interaction between anticoagulant strategy and access-site choice.


What causes alopecia areata?
The pathobiology of alopecia areata (AA), one of the most frequent autoimmune diseases and a major unsolved clinical problem, has intrigued dermatologists, hair biologists and immunologists for decades. Simultaneously, both affected patients and the physicians who take care of them are increasingly frustrated that there is still no fully satisfactory treatment. Much of this frustration results from the fact that the pathobiology of AA remains unclear, and no single AA pathogenesis concept can claim to be universally accepted. In fact, some investigators still harbour doubts whether this even is an autoimmune disease, and the relative importance of CD8(+) T cells, CD4(+) T cells and NKGD2(+) NK or NKT cells and the exact role of genetic factors in AA pathogenesis remain bones of contention. Also, is AA one disease, a spectrum of distinct disease entities or only a response pattern of normal hair follicles to immunologically mediated damage? During the past decade, substantial progress has been made in basic AA-related research, in the development of new models for translationally relevant AA research and in the identification of new therapeutic agents and targets for future AA management. This calls for a re-evaluation and public debate of currently prevalent AA pathobiology concepts. The present Controversies feature takes on this challenge, hoping to attract more skin biologists, immunologists and professional autoimmunity experts to this biologically fascinating and clinically important model disease.


Pain and depression in patients with newly diagnosed pancreas cancer.
To evaluate the prevalence of pain and depression, their correlation, and their effect on quality of life in patients with recently diagnosed adenocarcinoma of the pancreas (PC). Cross-sectional pain and psychosocial distress were assessed using validated instruments, including the Memorial Pain Assessment Card (MPAC), Beck Depression Inventory (BDI), Hopelessness Scale (BHS), and Functional Living Index-Cancer (FLIC). Patients were evaluated before their first operation for PC or first treatment with chemotherapy at a large tertiary-care cancer center. One hundred thirty patients with proven PC were studied: 83 before their operation and 47 before their first chemotherapy treatment. At the time of study entrance, 37% of patients had no pain and an additional 34% had pain that was mild or less severe. Only 29% of patients had moderate, strong, or severe pain. Chemotherapy patients reported significantly more intense pain than did preoperative patients (P = .02). Symptoms of depression were assessed using the BDI and BHS scales. A substantial minority of patients (38%) had BDI scores > or = 15, which suggests high levels of depressive symptoms. There was a significant correlation between increasing pain and depressive symptoms among those who experienced pain. Quality of life was assessed using the Weekly Activity Checklist (WAC) and the FLIC. Compared with patients who had no pain or mild pain, patients with moderate or greater pain had significantly impaired functional activity (P = .03) and poorer quality-of-life scores (P = .02) when compared with those with lesser degrees of pain. There were significant correlations between increasing pain and depression and between pain and depressive symptoms and impaired quality of life and function. Our results indicate that moderate or severe pain and symptoms of depression are not as prevalent in recently diagnosed PC patients as is generally believed. However, one third have inadequate pain control despite the use of oral analgesics. These patients can be identified by the use of a simple self-report instrument (the MPAC card). Quality of life and function are adversely affected by moderate or greater levels of perceived pain intensity. A simple and rapid assessment is possible and can identify high-risk patients in need of intervention that may improve quality of life.


Regulation of growth hormone signaling by selective estrogen receptor modulators occurs through suppression of protein tyrosine phosphatases.
Activation of the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 5 (STAT5) pathway by GH is terminated by the suppressors of cytokine signaling (SOCSs) and protein tyrosine phosphatases, Src homology 2 domain-containing protein tyrosine phosphatase (SHP)-1 and SHP-2. Based on our recent report that estrogen inhibits GH signaling by stimulating SOCS-2 expression, we investigated the effects of selective estrogen receptor modulators (SERMs) on GH signaling in human embryonic kidney (HEK293) and breast cancer (MDA-MB-231) cells expressing human GH receptor and estrogen receptor-alpha. 17beta-estradiol (E(2)) suppressed GH activation of a STAT5-responsive luciferase reporter and JAK2 phosphorylation in both cell models. 4-hydroxytamoxifen and raloxifene augmented these actions of GH in HEK293 cells but not breast cancer cells. SOCS-2 expression in both cell types was stimulated by E(2) but unaffected by SERMs. In HEK293 cells, SHP-1 was inhibited by raloxifene and 4-hydroxytamoxifen, whereas the latter additionally inhibited SHP-2. The phosphatases were unaffected by E(2). In breast cancer cells, phosphatase activity was not altered by SERMs or E(2). In summary, estrogen inhibited the JAK2/STAT5 signaling of GH and stimulated SOCS-2 expression in both HEK293 and breast cancer cells. By contrast, SERMs augmented GH signaling by reducing SHP activities in HEK293 cells and had no effect on both in breast cancer cells. We provide the first evidence for a novel mechanism regulating GH signaling, in which SERMs enhance GH activation of the JAK2/STAT5 pathway in a cell-type-dependent manner by attenuating protein tyrosine phosphatase activities.


The Isolation, Primacy, and Recency Effects Predicted by an Adaptive LTD/LTP Threshold in Postsynaptic Cells.
An item that stands out (is isolated) from its context is better remembered than an item consistent with the context. This isolation effect cannot be accounted for by increased attention, because it occurs when the isolated item is presented as the first item, or by impoverished memory of nonisolated items, because the isolated item is better remembered than a control list consisting of equally different items. The isolation effect is seldom experimentally or theoretically related to the primacy or the recency effects-that is, the improved performance on the first few and last items, respectively, on the serial position curve. The primacy effect cannot easily be accounted for by rehearsal in short-term memory because it occurs when rehearsal is eliminated. This article suggests that the primacy, the recency, and the isolation effects can be accounted for by experience-dependent synaptic plasticity in neural cells. Neurological empirical data suggest that the threshold that determines whether cells will show long-term potentiation (LTP) or long-term depression (LTD) varies as a function of recent postsynaptic activity and that synaptic plasticity is bounded. By implementing an adaptive LTP-LTD threshold in an artificial neural network, the various aspects of the isolation, the primacy, and the recency effects are accounted for, whereas none of these phenomena are accounted for if the threshold is constant. This theory suggests a possible link between the cognitive and the neurological levels.


One-year clinical results of the Italian diffuse/multivessel disease ABSORB prospective registry (IT-DISAPPEARS).
The present multicentre prospective study, IT-DISAPPEARS, was designed with the aim of evaluating early and long-term clinical outcomes of the Absorb BVS in patients with long coronary lesions and/or multivessel coronary artery disease. The aim of this article is to present the one-year clinical results of this study. Between November 2014 and January 2016, we enrolled 1,002 patients undergoing BVS implantation (long lesion [≥ 24 mm] of a single vessel in 80.4%, at least two BVS in two or three coronary vessels in 8.6% and both criteria in 11%). Clinical presentation was an acute coronary syndrome in 59.8% of patients, including ST-elevation myocardial infarction in 21.8%. The primary endpoint was the device-oriented composite endpoint (DOCE) of cardiac death, target vessel MI, and ischaemia-driven TLR at one year. We implanted 2,040 BVS according to a pre-specified technique. One-year follow-up was available in 956 patients (95.4%). The rate of DOCE was 9.9% (95 patients). Cardiac death occurred in five patients (0.5%), while target vessel MI and TLR each occurred in 45 (4.7%) patients. The one-year rates of all-cause death, non-fatal MI, and any revascularisation were 1.2%, 5.4%, and 10.9%, respectively. The rate of definite/probable scaffold thrombosis was 0.9%. This is the first study specifically investigating the Absorb technology in patients with a high atherosclerotic burden and multivessel disease. The mandatory adherence to a pre-specified implantation technique led to minimising the risk of device failure reported by other studies, in particular with respect to the rate of DOCE and scaffold thrombosis.


Nucleobases, nucleosides, and nucleotides: versatile biomolecules for generating functional nanomaterials.
The incorporation of biomolecules into nanomaterials generates functional nanosystems with novel and advanced properties, presenting great potential for applications in various fields. Nucleobases, nucleosides and nucleotides, as building blocks of nucleic acids and biological coenzymes, constitute necessary components of the foundation of life. In recent years, as versatile biomolecules for the construction or regulation of functional nanomaterials, they have stimulated interest in researchers, due to their unique properties such as structural diversity, multiplex binding sites, self-assembly ability, stability, biocompatibility, and chirality. In this review, strategies for the synthesis of nanomaterials and the regulation of their morphologies and functions using nucleobases, nucleosides, and nucleotides as building blocks, templates or modulators are summarized alongside selected applications. The diverse applications range from sensing, bioimaging, and drug delivery to mimicking light-harvesting antenna, the construction of logic gates, and beyond. Furthermore, some perspectives and challenges in this emerging field are proposed. This review is directed toward the broader scientific community interested in biomolecule-based functional nanomaterials.


CARD8 is a negative regulator for NLRP3 inflammasome, but mutant NLRP3 in cryopyrin-associated periodic syndromes escapes the restriction.
NLRP3 plays a role in sensing various pathogen components or stresses in the innate immune system. Once activated, NLRP3 associates with apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and procaspase-1 to form a large protein complex termed inflammasome. Although some investigators have proposed a model of NLRP3-inflammasome containing an adaptor protein caspase recruitment domain-containing protein 8 (CARD8), the role of this molecule remains obscure. This study aimed to clarify the interaction between CARD8 and wild-type NLRP3 as well as mutant forms of NLRP3 linked with cryopyrin-associated periodic syndromes (CAPS). In here HEK293 expression system, cells were transfected with the cDNAs for inflammasome components. Also used were peripheral blood mononuclear cells (PBMCs) and human monocyte-derived macrophages (HMDMs) from healthy volunteers. The interaction of CARD8 and NLRP3 was studied by immunoprecipitation. The effect of CARD8 expression on IL-1β secretion was assessed by ELISA. CARD8 knockdown experiments were carried out by transfection of the specific siRNA into HMDMs. In HEK293 cells, CARD8 interacted with wild-type NLRP3, but not with CAPS-associated mutant NLRP3. CARD8 significantly reduced IL-1β secretion from cells transfected with wild-type NLRP3, but not if they were transfected with mutant NLRP3. In addition, association of endogenously expressed CARD8 with NLRP3 was confirmed in resting PBMCs, and CARD8 knockdown resulted in higher amount of IL-1β secretion from HMDMs. Until specific stimuli activate NLRP3, CARD8 holds NLRP3, and is supposed to prevent activation by subtle stimuli. However, CAPS-associated mutant NLRP3 is unable to bind with CARD8, which might be relevant to the pathogenesis of CAPS.


Bonobos' saliva remaining on the pith of terrestrial herbaceous vegetation can serve as non-invasive wild genetic resources.
Evaluating the genetic diversity of natural populations of endangered species is important for conservation. Although the genetic analysis of wildlife usually requires collecting DNA non-invasively, the variety of non-invasive DNA sampling methods is limited for each species. We present a method to obtain DNA of an endangered species, the bonobo (Pan paniscus), in which the pith of the terrestrial herbaceous vegetation (THV) that they consumed was newly utilized. We investigated the (1) frequency of encountering remnant saliva on three types of THV pith; (2) concentrations of DNA in the saliva samples by the real-time quantitative PCR; and (3) rates of positive PCR, accurate genotyping, and allelic drop out by analyzing two autosomal microsatellite loci (D7s817 and D9s910). The number of remnant saliva samples was recorded by following the bonobo groups on a daily basis. The frequency of encountering DNA samples was higher in saliva samples than in fecal samples. More than half of the saliva samples remaining on two types of THV pith provided sufficient concentrations of bonobo DNA (> 200 pg/μl). Rates of positive PCR and accurate genotyping were high, and allelic drop out rate was low when the amount of template DNA was above 200 pg per reaction. Our results suggest that the remnants of bonobo saliva on the pith of THV are a potential resource for obtaining DNA, and better than other kinds of samples from the perspective of the abundant sampling opportunities.


The Impact of Low Energy Availability on Nonexercise Activity Thermogenesis and Physical Activity Behavior in Recreationally Trained Adults.
Energy availability describes the amount of dietary energy remaining for physiological functionality after the energy cost of exercise is deducted. The physiological and hormonal consequences of low energy availability (LEA) are well established, but the impact of LEA on physical activity behavior outside of exercise and, specifically, nonexercise activity thermogenesis (NEAT) has not been systematically examined. The authors conducted a secondary analysis of a repeated-measures crossover study in which recreationally trained young men (n = 6, 25 ± 1.0 years) underwent two 4-day conditions of LEA (15 kcal·kg fat-free mass-1 ·day-1) with and without endurance exercise (LEA + EX and LEA EX) and two energy-balanced control conditions (CON + EX and CON EX). The duration and intensity of physical activity outside of prescribed exercise were assessed using the SenseWear Pro3 armband. LEA did not alter NEAT (p = .41), nor time spent in moderate to vigorous (p = .20) and low-intensity physical activity (p = .17). However, time spent in low-intensity physical activity was lower in LEA + EX than LEA - EX (13.7 ± 0.3 vs. 15.2 ± 0.3 hr/day; p = .002). Short-term LEA does not seem to impact NEAT per se, but the way it is attained may impact physical activity behavior outside of exercise. As the participants expended similar amounts of energy during NEAT (900-1,300 kcal/day = 12.5-18.0 kcal·kg fat-free mass-1·day-1) and prescribed exercise bouts (15.0 kcal·kg fat-free mass-1·day-1), excluding it as a component of energy expenditure may skew the true energy available for physiological functionality in active populations.


Perioperative morbidity and long-term outcome of unilateral nephrectomy in feline kidney donors: 141 cases (1998-2013).
OBJECTIVE To evaluate the outcome associated with unilateral nephrectomy in feline kidney donors. DESIGN Retrospective case series. ANIMALS 141 cats. PROCEDURES Medical records of cats that underwent nephrectomy for renal donation were reviewed for information on signalment, date of renal donation, results of blood and urine analyses, infectious disease history, anesthetic protocols, intra- and postoperative complications, and postoperative analgesic protocols. Long-term follow-up data were obtained via client telephone interview and review of referring veterinarian medical records. RESULTS All donors were healthy young adult cats with a median age of 1.5 years (range, 0.8 to 2 years). No cats died or were euthanized during the perioperative period. Intraoperative complications occurred in 2 cats, and postoperative complications occurred in 17. Median time from nephrectomy to hospital discharge was 3.6 days (range, 2 to 8 days). Long-term follow-up information was available for 99 cats, with a median interval between nephrectomy and follow-up of 10 years (range, 0.25 to 15 years). Six cats had a history of urinary tract disease including stable chronic kidney disease (n = 3), acute kidney injury (2), and cystitis (1). Nine cats were dead at follow-up; death was attributed to chronic renal failure in 2 and acute ureteral obstruction in 4. CONCLUSIONS AND CLINICAL RELEVANCE Feline donor nephrectomy had an acceptably low perioperative morbidity in this series. Most cats (84%) for which follow-up information was available had no associated long-term effects. However, a small subset (7%) developed renal insufficiency or died of urinary tract disease.


Incorporation of Time-of-Flight Information Reduces Metal Artifacts in Simultaneous Positron Emission Tomography/Magnetic Resonance Imaging: A Simulation Study.
This study aimed to describe and evaluate the influence of time-of-flight (TOF) information on metal artifact reduction in positron emission tomography (PET) image quality in clinical simultaneous PET/magnetic resonance (MR) scanning. A total of 7 patients with various malignant tumors were included and underwent a PET/MR examination after standard PET/computed tomography. Baseline TOF and non-TOF PET images were reconstructed. Next, the TOF and non-TOF PET reconstructions were repeated after the introduction of artificial signal voids in the attenuation map to simulate metal artifacts of various sizes in a range of locations. Three different sizes of signal voids were inserted in the attenuation maps for each location of interest: over the maxilla, humeral head, chest, sternum, thoracic and lumbar spine, as well as the femoral head to replicate clinically relevant metal artifacts. The reconstructed images with the artifacts were then compared with the baseline reconstructed images. The mean percentage error in a region of interest surrounding the simulated artifact was used to compare between TOF and non-TOF images. Further comparison between TOF and non-TOF images was performed using histogram analysis. In all cases, the mean percentage error in a region of interest surrounding the simulated artifact was reduced when TOF information was included in the reconstruction. The inclusion of TOF also changes the distribution of smaller errors away from the origin of the artifact. In some anatomical regions, an increase in the number of small errors was noted with TOF, although the differences with non-TOF were minimal. Positron emission tomographic imaging benefits from the integration of TOF information in simultaneous PET/MR. The inclusion of TOF information in simultaneous PET/MR imaging reduces errors related to metal artifacts at the site of the artifact.


Limited HIV-1 superinfection in seroconverters from the CAPRISA 004 Microbicide Trial.
HIV-1 superinfection (SI) occurs when an infected individual acquires a distinct new viral strain. The rate of superinfection may be reflective of the underlying HIV risk in a population. The Centre for the AIDS Programme of Research in South Africa (CAPRISA) 004 clinical trial demonstrated that women who used a tenofovir-containing microbicide gel had lower rates of HIV infection than women using a placebo gel. Women who contracted HIV-1 during the trial were screened for the occurrence of superinfection by next-generation sequencing of the viral gag and env genes. There were two cases (one in each trial arm) of subtype C superinfection identified from the 76 women with primary infection screened at two time points (rate of superinfection, 1.5/100 person-years). Both women experienced a >0.5-log increase in viral load during the window when superinfection occurred. The rate of superinfection was significantly lower than the overall primary HIV incidence in the microbicide trial (incidence rate ratio [IRR], 0.20; P=0.003). The women who seroconverted during the trial reported a significant increase in sexual contact with their stable partner 4 months after seroconversion (P<0.001), which may have lowered the risk of superinfection in this population. The lower frequency of SI compared to the primary incidence is in contrast to a report from a general heterosexual African population but agrees with a study of high-risk women in Kenya. A better understanding of the rate of HIV superinfection could have important implications for ongoing HIV vaccine research.


The Fast Component of Visual Field Decay Rate Correlates With Disc Rim Area Change Throughout the Entire Range of Glaucomatous Damage.
We investigated the longitudinal relationships between the changes in neuroretinal rim area (RA) and the slow (SC) and fast (FC) components of visual field (VF) decay at various stages of glaucoma. We divided 465 eyes of 338 patients into glaucoma suspect, and preperimetric, early, and moderate/advanced glaucoma. All patients had a minimum of 3 confocal scanning laser ophthalmoscopic examinations and 4 VF tests with follow-up of 4 or more years. A pointwise exponential regression was used to perform trend analyses on thresholds at each VF test location, which was partitioned into SC and FC. A mixed effects linear model was used to explore the associations of RA change with mean deviation (MD), visual field index (VFI), SC, and FC. Decreased RA was associated with lower mean threshold sensitivities of FC regardless of baseline severity of glaucoma (P ≤ 0.03). The mean threshold sensitivities in SC were not correlated with RA change at any stage. Decreased RA was correlated with worse MD in preperimetric, early, and moderate/advanced glaucoma (P < 0.05). Decreased RA was correlated with worse VFI in preperimetric and early glaucoma only (P ≤ 0.04). A decrease in rim area was significantly correlated with the fast VF component regardless of the baseline severity of glaucoma. Mean deviation and VFI correlated with change of rim area only in certain stages of glaucoma. The identification of the fast component seems a more robust and useful measure of glaucomatous change than MD or VFI.


Catalysis in a Cationic Coordination Cage Using a Cavity-Bound Guest and Surface-Bound Anions: Inhibition, Activation, and Autocatalysis.
The Kemp elimination (reaction of benzisoxazole with base to give 2-cyanophenolate) is catalyzed in the cavity of a cubic M8L12 coordination cage because of a combination of (i) benzisoxazole binding in the cage cavity driven by the hydrophobic effect, and (ii) accumulation of hydroxide ions around the 16+ cage surface driven by ion-pairing. Here we show how reaction of the cavity-bound guest is modified by the presence of other anions which can also accumulate around the cage surface and displace hydroxide, inhibiting catalysis of the cage-based reaction. Addition of chloride or fluoride inhibits the reaction with hydroxide to the extent that a new autocatalytic pathway becomes apparent, resulting in a sigmoidal reaction profile. In this pathway the product 2-cyanophenolate itself accumulates around the cationic cage surface, acting as the base for the next reaction cycle. The affinity of different anions for the cage surface is therefore 2-cyanophenolate (generating autocatalysis) > chloride > fluoride (which both inhibit the reaction with hydroxide but cannot deprotonate the benzisoxazole guest) > hydroxide (default reaction pathway). The presence of this autocatalytic pathway demonstrates that a reaction of a cavity-bound guest can be induced with different anions around the cage surface in a controllable way; this was confirmed by adding different phenolates to the reaction, which accelerate the Kemp elimination to different extents depending on their basicity. This represents a significant step toward the goal of using the cage as a catalyst for bimolecular reactions between a cavity-bound guest and anions accumulated around the surface.


Polydimethylsiloxane-based optical waveguides for tetherless powering of floating microstimulators.
Neural electrodes and associated electronics are powered either through percutaneous wires or transcutaneous powering schemes with energy harvesting devices implanted underneath the skin. For electrodes implanted in the spinal cord and the brain stem that experience large displacements, wireless powering may be an option to eliminate device failure by the breakage of wires and the tethering of forces on the electrodes. We tested the feasibility of using optically clear polydimethylsiloxane (PDMS) as a waveguide to collect the light in a subcutaneous location and deliver to deeper regions inside the body, thereby replacing brittle metal wires tethered to the electrodes with PDMS-based optical waveguides that can transmit energy without being attached to the targeted electrode. We determined the attenuation of light along the PDMS waveguides as 0.36 ± 0.03 ?? dB / cm and the transcutaneous light collection efficiency of cylindrical waveguides as 44 % ± 11 % by transmitting a laser beam through the thenar skin of human hands. We then implanted the waveguides in rats for a month to demonstrate the feasibility of optical transmission. The collection efficiency and longitudinal attenuation values reported here can help others design their own waveguides and make estimations of the waveguide cross-sectional area required to deliver sufficient power to a certain depth in tissue.


Detection of clinical mastitis with sensor data from automatic milking systems is improved by using decision-tree induction.
The objective was to develop and validate a clinical mastitis (CM) detection model by means of decision-tree induction. For farmers milking with an automatic milking system (AMS), it is desirable that the detection model has a high level of sensitivity (Se), especially for more severe cases of CM, at a very high specificity (Sp). In addition, an alert for CM should be generated preferably at the quarter milking (QM) at which the CM infection is visible for the first time. Data were collected from 9 Dutch dairy herds milking automatically during a 2.5-yr period. Data included sensor data (electrical conductivity, color, and yield) at the QM level and visual observations of quarters with CM recorded by the farmers. Visual observations of quarters with CM were combined with sensor data of the most recent automatic milking recorded for that same quarter, within a 24-h time window before the visual assessment time. Sensor data of 3.5 million QM were collected, of which 348 QM were combined with a CM observation. Data were divided into a training set, including two-thirds of all data, and a test set. Cows in the training set were not included in the test set and vice versa. A decision-tree model was trained using only clear examples of healthy (n=24,717) or diseased (n=243) QM. The model was tested on 105 QM with CM and a random sample of 50,000 QM without CM. While keeping the Se at a level comparable to that of models currently used by AMS, the decision-tree model was able to decrease the number of false-positive alerts by more than 50%. At an Sp of 99%, 40% of the CM cases were detected. Sixty-four percent of the severe CM cases were detected and only 12.5% of the CM that were scored as watery milk. The Se increased considerably from 40% to 66.7% when the time window increased from less than 24h before the CM observation, to a time window from 24h before to 24h after the CM observation. Even at very wide time windows, however, it was impossible to reach an Se of 100%. This indicates the inability to detect all CM cases based on sensor data alone. Sensitivity levels varied largely when the decision tree was validated per herd. This trend was confirmed when decision trees were trained using data from 8 herds and tested on data from the ninth herd. This indicates that when using the decision tree as a generic CM detection model in practice, some herds will continue having difficulties in detecting CM using mastitis alert lists, whereas others will perform well.


UM171 Expansion of Cord Blood Improves Donor Availability and HLA Matching For All Patients, Including Minorities.
Cord blood (CB) stem cell transplantation offers a greater tolerance to HLA mismatches compared to adult-derived stem cell transplants (i.e., bone marrow or peripheral blood stem cells). Indeed, 4/6 or 5/8 HLA-matched CB transplantations are regularly performed for patients lacking a matched unrelated donor. Unfortunately, most banked CB units contain a stem cell dose that is too small to treat adult patients, resulting in only 4% to 5% of available CB units offering an adequate cell dose for prompt engraftment for adult patients. Ex vivo stem cell expansion appears to be an attractive strategy to circumvent this cell dose issue, while also enabling the selection of better HLA-matched CB units. In this study, we retrospectively performed HLA matching simulations to assess how the minimal cell content requirements associated with UM171 CB expansion may improve usability of existing CB unit inventories and donor availability for patients of different races and ethnicities. We analyzed a dataset of 58,971 adults for whom a donor search was initiated through the National Marrow Donor Program Be The Match registry against 142,942 CB units from major U.S. public CB banks listed on the Be The Match registry. Our results show that by enabling selection of smaller CB units, UM171-expanded CB transplantation increases donor availability from 72% to 84% for all patients compared to single unmanipulated CB transplantation. Furthermore, the low cell dose criteria for UM171-expanded CB also increases donor availability compared to double CB transplantation, while enabling better HLA matching between donor and recipient. UM171 expanded CB appears particularly beneficial for racial and ethnic minority patients as CB availability increases from 53% to 78% for African Americans, from 66% to 85% for Hispanics, and from 68% to 84% for Asians and Pacific Islanders, compared to single unmanipulated CB transplantation. In addition, UM171 expansion dramatically improves usability of CB units currently in inventories, as only 4.3% and 0.6% of banked CBs have sufficient cell doses for a 70 kg and 100 kg patient, respectively. UM171 raises this proportion to 53.8% and 20.2%, respectively, making CB banks potentially more cost effective. In conclusion, UM171 expansion allows the use of smaller CB units while also improving access to transplantation for racial and ethnic minorities.


Aphasia in Neurology Practice: A Survey about Perceptions and Practices.
Aphasia is one of those clinical conditions, where the role of affiliated professionals, mainly speech language pathologists (SLPs) is substantial in diagnostic assessments, therapy, and rehabilitation. There is no study to focus on neurologists, with respect to their perceptions and practices about aphasia, the disease, as well as the profession of SLP. To reach out to the neurologist community in India and learn about their perceptions and practices about the nature of the ailment and role of speech language therapy (SLT). Our premise was that observations and inferences from a questionnaire-based survey will be subsequently helpful in planning educational activities targeted to neurologists with more focus on specific gaps in perceptions and practices. Three neurologists and two SLPs collaboratively developed the questionnaire. The aim was to probe the issues which were likely to have a bearing upon optimum service delivery to persons with aphasia by a dyad of neurologist and SLP. The survey was set in "Google Forms" and sent by "WhatsApp" and email to approximately 500 practicing neurologists in India. We employed a nonprobability sampling design for ease of administration with a combination of "chunk sampling" and "snowball sampling." Telephonic reminders were made to almost all. We received 100 responses. The mean age of respondents was 50.64 (SD +/- 12.60) with a range of 28-78 years. The mean number of years of experience as a neurophysician was 19.88 (SD. +/- 12.72) with range of 1-47 years. Females were only 8%. Apparently, the proportion of neurologists working in large corporate and large public sector institutions from tier one and tier two cities was higher, who are more likely to have SLP and related rehabilitation facilities in their institutions and hence harbor more conducive attitudes to SLT in aphasia. The ground reality from tier three cities and small private and public sector hospital and solo practitioners may be somewhat worse than this. Many responses were in conformity with facts and in tune with desirable attitudes as per guidelines like aphasia being a detrimental factor in stroke recovery, doing assessment of handedness, paying attention to neuroimaging correlations and associated cognitive functions, not resorting to unnecessary pharmacotherapy, being aware about efficacy of SLT, and fairly good chances of recovery. However, many more answers highlighted a need for emphasis in Continuing Medical Education like not being aware about community burden of aphasia in comparison to a few better known neurological diseases, not paying attention to psychosocial aspects apart from biological ones in assessment and rehabilitation, not using a standardized and validated battery, not confidant about role of SLT in chronic stable aphasia and need for longer and intensive therapy, and being unconcerned for the value of advocacy for aphasia, like the role of Self-Help Groups. The thrust areas, pertaining to gaps in perception and practices identified through this study, can be viewed as "an in-time input." We hope that changes in some of the perceptions and practices can be attained through an emphasis on education and training at multiple levels right from the undergraduate to the practicing physicians. A few more themes and domains will need advocacy actions targeted to different stakeholders.


A digital heterodyne laser interferometer for studying cochlear mechanics.
Laser interferometry is the technique of choice for studying the smallest displacements of the hearing organ. For low intensity sound stimulation, these displacements may be below 1 nm. This cannot be reliably measured with other presently available techniques in an intact organ of Corti. In a heterodyne interferometer, light is projected against an object of study and motion of the target along the optical axis causes phase and frequency modulations of the back-reflected light. To recover object motion, the reflected light is made to interfere with a reference beam of artificially altered frequency, producing a beating signal. In conventional interferometers, this carrier signal is demodulated with analog electronics. In this paper, we describe a digital implementation of the technique, using direct carrier sampling. In order to obtain the necessary reference signal for demodulation we introduce an additional third light path. Together, this results in lower noise and reduces the cost of the system. Within the hearing organ, different structures may move in different directions. It is therefore necessary to precisely measure the angle of incidence of the laser light, and to precisely localize the anatomical structure where the measurement is performed. Therefore, the interferometer is integrated with a laser scanning confocal microscope that permits us to map crucial morphometric parameters in each experiment. We provide key construction parameters and a detailed performance characterization. We also show that the system accurately measures the diminutive vibrations present in the apical turn of the cochlea during low-level sound stimulation.


Exercise alters pain sensitivity in Gulf War veterans with chronic musculoskeletal pain.
Since returning from the Persian Gulf, nearly 100,000 veterans of the first Gulf War (GVs) have reported numerous symptoms with no apparent medical explanation. A primary complaint of these individuals is chronic musculoskeletal pain (CMP). CMP symptoms in GVs are similar to those reported by patients with fibromyalgia (FM), but have not received equivalent scientific attention. Exercise research in CMP patients suggests that acute exercise may exacerbate pain while chronic exercise can reduce pain and improve other symptoms. However, the influence of exercise on GVs with CMP is largely unexplored. This study examined the impact of an acute bout of exercise on pain sensitivity in GVs with CMP. Thirty-two GVs (CMP, n = 15; Control, n = 17) were recruited to complete a series of psychophysical assessments to determine pain sensitivity to heat and pressure stimuli before and after exercise. In response to heat-pain stimuli, GVs with CMP reported higher pain intensity and affect ratings than healthy GVs and exhibited a significant increase in ratings following exercise. GVs with CMP rated exercise as more painful and effortful and were generally more sensitive to heat-pain stimuli than healthy GVs. These results are similar to what has been reported for acute exercise in patients with FM. Gulf War veterans with CMP perceive exercise as more painful and effortful than healthy GVs and experience increased pain sensitivity following exercise. These results suggest that similar abnormalities in central nervous system processing of nociceptive information documented in FM may also be occurring in GVs with CMP.


Axonal modulation of myelin gene expression in the peripheral nerve.
Myelin gene expression (P0, MBP, P2, and MAG) was investigated during Wallerian degeneration and in the presence or absence of subsequent axonal regeneration and remyelination. The steady state levels of mRNA and protein were assessed in the crushed or permanently transected rat sciatic nerve at 0, 1, 4, 7, 10, 12, 14, 21, and 35 days after injury. The mRNA and protein steady state levels of the myelin specific genes, P0 and the MBPs, decreased to low yet detectable levels during Wallerian degeneration and returned to normal levels with subsequent axonal regeneration. The steady state level of P2 protein also followed a similar pattern of expression. The steady state level of MAG mRNA decreased to undetectable levels by 4 days of injury in the permanently transected nerve. After crush injury, re-expression of MAG to levels comparable to those of normal nerves preceded that of P2 by 2 days and that of P0 and the MBPs by 3 weeks during axonal regeneration and remyelination. These results support the proposed roles for MAG in the formation of initial Schwann cell-axonal contact required for myelin assembly, for P2 in fatty acid transport during myelination, and for P0 and the MBPs in the maintenance of the integrity and compactness of the myelin sheath. In addition, these results indicate that the expression of the myelin specific genes, P0 and MBP, is constitutive and that the level of myelin specific mRNAs is modulated by axonal contact and myelin assembly.


An active learning curriculum improves fellows' knowledge and faculty teaching skills.
Traditional didactic lectures are the mainstay of teaching for graduate medical education, although this method may not be the most effective way to transmit information. We created an active learning curriculum for Brigham and Women's Hospital (BWH) gastroenterology fellows to maximize learning. We evaluated whether this new curriculum improved perceived knowledge acquisition and knowledge base. In addition, our study assessed whether coaching faculty members in specific methods to enhance active learning improved their perceived teaching and presentation skills. We compared the Gastroenterology Training Exam (GTE) scores before and after the implementation of this curriculum to assess whether an improved knowledge base was documented. In addition, fellows and faculty members were asked to complete anonymous evaluations regarding their learning and teaching experiences. Fifteen fellows were invited to 12 lectures over a 2-year period. GTE scores improved in the areas of stomach (p<0.001), general gastroenterology (p=0.005), esophagus (p<0.001), and small bowel (p=0.001), and the total score (p=0.001) between pre- and postimplementation of the active learning curriculum. Scores in hepatology, as well as biliary and pancreatic study, showed a trend toward improvement (p>0.05). All fellows believed the lectures were helpful, felt more prepared to take the GTE, and preferred the interactive format to traditional didactic lectures. All lecturers agreed that they acquired new teaching skills, improved teaching and presentation skills, and learned new tools that could help them teach better in the future. An active learning curriculum is preferred by GI fellows and may be helpful for improving transmission of information in any specialty in medical education. Individualized faculty coaching sessions demonstrating new ways to transmit information may be important for an individual faculty member's teaching excellence.


Identification of urinary peptide biomarkers associated with rheumatoid arthritis.
Early diagnosis and treatment of rheumatoid arthritis are associated with improved outcomes but current diagnostic tools such as rheumatoid factor or anti-citrullinated protein antibodies have shown limited sensitivity. In this pilot study we set out to establish a panel of urinary biomarkers associated with rheumatoid arthritis using capillary electrophoresis coupled to mass spectrometry. We compared the urinary proteome of 33 participants of the Scottish Early Rheumatoid Arthritis inception cohort study with 30 healthy controls and identified 292 potential rheumatoid arthritis-specific peptides. Amongst them, 39 were used to create a classifier model using support vector machine algorithms. Specific peptidic fragments were differentially excreted between groups; fragments of protein S100-A9 and gelsolin were less abundant in rheumatoid arthritis while fragments of uromodulin, complement C3 and fibrinogen were all increasingly excreted. The model generated was subsequently tested in an independent test-set of 31 samples. The classifier demonstrated a sensitivity of 88% and a specificity of 93% in diagnosing the condition, with an area under the receiver operating characteristic curve of 0.93 (p<0.0001). These preliminary results suggest that urinary biomarkers could be useful in the early diagnosis of rheumatoid arthritis. Further studies are currently being undertaken in larger cohorts of patients with rheumatoid arthritis and other athridities to assess the potential of the urinary peptide based classifier in the early detection of rheumatoid arthritis.


Association of the intestinal fatty acid-binding protein Ala54Thr polymorphism and abdominal adipose tissue in African-American and Caucasian women.
Genetic variants in the intestinal fatty acid-binding protein-2 (FABP2) gene have been associated with body composition phenotypes. We examined the association between the Ala(54)Thr variant in the FABP2 gene and levels of visceral (VAT) and sc (SAAT) abdominal fat in a group of 223 premenopausal African-American (n = 103) and Caucasian (n = 120) women. Subjects were genotyped for the marker. In addition, body composition was assessed by dual energy x-ray absorptiometry, and VAT was determined from a single computed tomography scan. The frequency of the Thr mutant allele did not differ significantly by ethnic group. After adjusting for total body fat, total abdominal adipose tissue (TAT) and SAAT were significantly lower in carriers of either one or two copies of the mutant Thr allele (P < 0.01). There was no association between total fat mass or VAT and the FABP2 polymorphism. Separate analyses by ethnic group showed that the association between the polymorphism and TAT and SAAT was observed in Caucasian (P < 0.01), but not in African-American (not significant), women. We conclude that women carriers of the FABP2 Thr allele have lower TAT and SAAT than noncarriers of the mutation. This association is present in Caucasian, but not in African-American, women.


Dietary patterns and practices in rural eastern Uganda: Implications for prevention and management of type 2 diabetes.
The burden of type 2 diabetes in Sub-Saharan Africa is projected to double by 2040, partly attributable to rapidly changing diets. In this paper, we analysed how community members in rural Uganda understood the concept of a healthy or unhealthy diet, food preparation and serving practices to inform the process of facilitating knowledge and skill necessary for self-management and care for type 2 diabetes. This was a qualitative study involving 20 focus group discussions and eight in-depth interviews among those at risk, patients with type 2 diabetes and the general community members without diabetes mellitus. Data was coded and entered into Atlas ti version 7.5.12 and interpreted using thematic analysis. We identified three main themes, which revealed, the perceptions on food and diet concerning health; the social dimensions of food and influence on diet practices; and food as a gendered activity. Participants noted that eating and cooking practices resulted in unhealthy diets. Their practices were affected by beliefs, poverty and food insecurity. Women determined which foods to prepare, but men prepared only some of the foods such as delicacies like a rice dish "pilau." New commercial and processed foods were increasingly available and consumed even in rural areas. Participants linked signs and symptoms of illness to diet as they narrated changes from past to current food preparation behaviours. Their view of overweight and obesity was also gendered and linked to social status. Participants' perception of disease influenced by diet was similar among those with and without type 2 diabetes, and those at risk. People described what is a healthy diet was as recommended by the health workers, but stated that their practices differed greatly from their knowledge. There was high awareness about healthy and balanced diets, but food is entrenched within social and gendered paradigms, which are slowly changing. Social and gender dimensions of food will need to be addressed through interventions in communities to promote change on a society level.


Drug treatment of microsporidiosis.
Microsporidia are ubiquitous organisms that are emerging pathogens in humans. These are most likely zoonotic and/or waterborne infections. In the immunosuppressed host, such as those treated with immunosuppressive drugs or infected with human immunodeficiency virus particularly at advanced stages of the disease, microsporidia can produce a wide range of clinical diseases. The most common manifestation is gastrointestinal tract infection; however, encephalitis, ocular infection, sinusitis, myositis and disseminated infection have also been described. In addition, these organisms have been reported in immune competent individuals. Multiple genera are involved in these infections and different organisms can result in distinct clinical pictures. Differences in clinical and parasitologic response to various therapeutic agents have emerged from clinical, as well as in vitro and in vivo studies. Currently there are no precisely defined guidelines for the optimal treatment of microsporidial infections. This article reviews the available data on compounds with in vitro activity and/or in vivo efficacy for microsporidial infections. Copyright 2000 Harcourt Publishers Ltd.


Functional analysis of the rat insulin-like growth factor I gene and identification of an IGF-I gene promoter.
Insulin-like growth factor I (IGF-I) mediates many of the systemic growth-promoting effects of growth hormone and also functions as a locally acting growth stimulator. In mammals, IGF-I gene expression is complicated, as the gene is transcribed and processed into multiple mRNAs (ranging in length from less than 1 to nearly 7.5 kb) that encode at least two protein precursors. As a step toward understanding the regulation of IGF-I, we report the complete organization of the rat IGF-I gene, including identification of the structural determinants for all IGF-I mRNA species, and an initial functional analysis of its promoters. The gene is composed of 6 exons distributed over nearly 80 kb of chromosomal DNA and is structurally heterogeneous. Several transcription start sites were identified within IGF-I exons 1 and 2, adjacent to presumptive promoters 1 and 2, respectively, and at least three polyadenylation sites were mapped to exon 6. To test promoter function, fusion genes were constructed linking fragments of IGF-I DNA to a reporter plasmid. Chimeric genes containing at least 395 bp of DNA from the 5'-flanking region of exon 1 enhanced luciferase activity after transfection into the IGF-I-producing SK-N-MC cell line, while fusion plasmids containing up to 1,300 bp of DNA from the 5'-flanking region of exon 2 were inactive. Relative levels of IGF-I mRNAs containing exons 1 or 2 varied among different rat tissues, although in response to acute or chronic growth hormone treatment both classes of transcripts were induced coordinately in rat liver. These observations represent the first thorough characterization of a mammalian IGF-I gene, and provide a starting point for defining the mechanisms by which growth hormone and other trophic factors regulate IGF-I gene expression.


Rose Bengal-photosensitized oxidation of 4-thiothymidine in aqueous medium: evidence for the reaction of the nucleoside with singlet state oxygen.
The photoreactivity of 4-thiothymidine (S(4)TdR) under visible light in the presence of Rose Bengal (RB), acting as a photosensitizer, was investigated in aqueous solutions at pH 7 and 12, using UV-vis, FTIR-ATR and (1)H-NMR spectroscopic techniques, time resolved absorption spectroscopy and electrospray ionization mass spectrometry (ESI-MS). Evidence for the generation of thymidine (TdR) as the main product, after one hour of irradiation, was obtained from UV-Vis data, that suggested 4-thiothymidine photodegradation to be faster at basic pH, and confirmed by FTIR-ATR and (1)H-NMR data. Clues for the presence of a further product, likely corresponding to a dimeric form of S(4)TdR, were obtained from the latter techniques. Besides indicating the presence of thymidine, the ESI-MS and MS/MS spectra of the reaction mixtures enabled the identification of the additional product as a S-S bridged covalent dimer of 4-thiothymidine. The concentration of the dimeric species could be estimated with the aid of (1)H-NMR data and was found to be lower than that of thymidine in pH 7 reaction mixtures and almost negligible in the pH 12 ones. From a mechanistic point of view, time-resolved absorption spectroscopy measurements provided direct evidence that the formation of the two products cannot be ascribed to a photoinduced electron transfer involving S(4)TdR and the excited triplet state of RB. Rather, their generation can be interpreted as the result of a bimolecular reaction occurring between singlet state oxygen ((1)O2), photogenerated by RB, and S(4)TdR, as demonstrated by the direct detection of (1)O2 through IR luminescence spectroscopy. More specifically, a sequential reaction pathway, consisting in the generation of an electrophilic hydroxylated form of S(4)TdR and its subsequent, rapid reaction with S(4)TdR, was hypothesized to explain the presence of the S-S bridged covalent dimer of 4-thiothymidine in the reaction mixtures. The described processes make S(4)TdR an interesting candidate in the role of molecular probe for the detection of (1)O2 under different pH conditions.


Principles of designing a cohort study in orthopaedics.
A well-designed and executed prospective cohort study can provide high-quality evidence in the evaluation of the effectiveness of surgical interventions. In designing a cohort study to evaluate orthopaedic interventions, it is important to recognize the limitations of the design as well as the methodological features that can be incorporated to strengthen the validity of the conclusions. In this article, we discuss the importance of the appropriate selection of participants for a control group, the management of confounders, the selection of outcomes with established measurement properties (reliability, validity, and sensitivity to change), the blinded assessment of outcomes, and the impact of nonparticipants and patients lost to follow-up.


Control of cellular GADD34 levels by the 26S proteasome.
GADD34, the product of a growth arrest and DNA damage-inducible gene, is expressed at low levels in unstressed cells. In response to stress, the cellular content of GADD34 protein increases and, on termination of stress, rapidly declines. We investigated the mechanisms that control GADD34 levels in human cells. GADD34 proteins containing either an internal FLAG or a C-terminal green fluorescent protein epitope were degraded at rates similar to endogenous GADD34. However, the addition of epitopes at the N terminus or deletion of N-terminal sequences stabilized GADD34. N-terminal peptides of GADD34, either alone or fused to heterologous proteins, exhibited rapid degradation similar to wild-type GADD34, thereby identifying an N-terminal degron. Deletion of internal PEST repeats had no impact on GADD34 stability but modulated the binding and activity of protein phosphatase 1. Proteasomal but not lysosomal inhibitors enhanced GADD34 stability and eukaryotic initiation factor 2alpha (eIF-2alpha) dephosphorylation, a finding consistent with GADD34's role in assembling an eIF-2alpha phosphatase. GADD34 was polyubiquitinated, and this modification enhanced its turnover in cells. A stabilized form of GADD34 promoted the accumulation and aggregation of the mutant cystic fibrosis transmembrane conductance regulator (CFTRDeltaF508), highlighting the physiological importance of GADD34 turnover in protein processing in the endoplasmic reticulum and the potential impact of prolonged GADD34 expression in human disease.


Metabolic engineering of glycine betaine synthesis: plant betaine aldehyde dehydrogenases lacking typical transit peptides are targeted to tobacco chloroplasts where they confer betaine aldehyde resistance.
Certain higher plants synthesize and accumulate glycine betaine, a compound with osmoprotectant properties. Biosynthesis of glycine betaine proceeds via the pathway choline-->betaine aldehyde-->glycine betaine. Plants such as tobacco (Nicotiana tabacum L.) which do not accumulate glycine betaine lack the enzymes catalyzing both reactions. As a step towards engineering glycine betaine accumulation into a non-accumulator, spinach and sugar beet complementary-DNA sequences encoding the second enzyme of glycine-betaine synthesis (betaine aldehyde dehydrogenase, BADH, EC 184.108.40.206) were expressed in tobacco. Despite the absence of a typical transit peptide, BADH was targeted to the chloroplast in leaves of transgenic plants. Levels of extractable BADH were comparable to those in spinach and sugar beet, and the molecular weight, isoenzyme profile and Km for betaine aldehyde of the BADH enzymes from transgenic plants were the same as for native spinach or sugar beet BADH. Transgenic plants converted supplied betaine aldehyde to glycine betaine at high rates, demonstrating that they were able to transport betaine aldehyde across both the plasma membrane and the chloroplast envelope. The glycine betaine produced in this way was not further metabolized and reached concentrations similar to those in plants which accumulate glycine betaine naturally. Betaine aldehyde was toxic to non-transformed tobacco tissues whereas transgenic tissues were resistant due to detoxification of betaine aldehyde to glycine betaine. Betaine aldehyded ehydrogenase is therefore of interest as a potential selectable marker, as well as in the metabolic engineering of osmoprotectant biosynthesis.


Undergraduate nursing students' transformational learning during clinical training.
Undergraduate nursing students encounter patients at the end of life during their clinical training. They need to confront dying and death under supportive circumstances in order to be prepared for similar situations in their future career. To explore undergraduate nursing students' descriptions of caring situations with patients at the end of life during supervised clinical training. A qualitative study using the critical incident technique was chosen. A total of 85 students wrote a short text about their experiences of caring for patients at the end of life during their clinical training. These critical incident reports were then analysed using deductive and inductive content analysis. The theme 'students' transformational learning towards becoming a professional nurse during clinical training' summarises how students relate to patients and relatives, interpret the transition from life to death, feel when caring for a dead body and learn end-of-life caring actions from their supervisors. As a preparation for their future profession, students undergoing clinical training need to confront death and dying while supported by trained supervisors and must learn how to communicate about end-of-life issues and cope with emotional stress and grief.


From post-traumatic stress disorder to absolute dependence in an intensive care unit: reflections on a clinical account.
This paper tells the story of one man's experience of terrifying hallucinations and nightmares in an intensive care unit (ICU), drawing attention to the reality that intensive care treatment induces emotional suffering severe enough to be identified as post-traumatic stress disorder (PTSD). A body of international research, confirmed by South African studies, links life-saving critical care to symptoms which qualify for secondary psychiatric diagnosis including of post-traumatic stress. Risk factors include pre-ICU comorbid psychopathology. Early on in the clinical encounter with the patient in this paper it emerged that he bore the scars of another trauma. He had been a soldier. Recounting the terror he experienced when he was being weaned off mechanical ventilation evoked memories of his military history. Paradoxically, these shifted the focus away from the symptoms of PTSD, to make the helplessness and dependency of ICU patients more visible. This patient's clinical account and patient experiences in other studies reveal the relational vulnerability of ICU patients. In as much as experiences of ICU treatment can be terrifying, the non-response of carers distresses patients. This interplay of wounding and care provides a starting point from which to explore how we account for the neglect of relational care that is a recurring theme in medical contexts, without blaming the carers. These questions find resonance in a South African novel to which the paper refers. A novel about war and trauma movingly portrays the internal conflict of the central character, a nurse and her quest not to care, as a defence against vulnerability. In these ways writing about the relational vulnerability of patients opened up questions about the social and institutional context of carer vulnerability.


Repetitive Peripheral Magnetic Stimulation With Intensive Swallowing Rehabilitation for Poststroke Dysphagia: An Open-Label Case Series.
The purpose of this pilot study was to determine the safety and feasibility of a six-day protocol of in-hospital repetitive peripheral magnetic stimulation combined with intensive swallowing rehabilitation (rPMS-ISR) for poststroke dysphagia. The subjects were eight patients with dysphagia caused by bilateral cerebral infarction (age: 62-70; time from onset of stroke: 27-39 months). rPMS was applied to the suprahyoid muscles, at strength set at 90% of the minimal intensity that elicited pain with a parabolic coil. One train of stimuli comprised 20 Hz for 3 sec followed by 27-sec rest. A single session included delivery of repetitive 20 trains of stimuli over 10 min, followed by 20 min of swallowing rehabilitation. Each patient received this combination treatment twice daily, morning and afternoon, over six consecutive days. Swallowing function was evaluated before and after intervention. rPMS-ISR induced significant improvement in swallowing ability, laryngeal elevation delay time, penetration aspiration scale, and swallowing quality of life (p < 0.01), but had no significant effect on the functional oral intake scale. The six-day in-hospital RPMS-ISR protocol seems safe and feasible for poststroke patients with dysphagia. The combination protocol improved swallowing function. Further larger studies are needed to confirm its efficacy.


Non-communicable diseases and global health governance: enhancing global processes to improve health development.
This paper assesses progress in the development of a global framework for responding to non-communicable diseases, as reflected in the policies and initiatives of the World Health Organization (WHO), World Bank and the UN: the institutions most capable of shaping a coherent global policy. Responding to the global burden of chronic disease requires a strategic assessment of the global processes that are likely to be most effective in generating commitment to policy change at country level, and in influencing industry behaviour. WHO has adopted a legal process with tobacco (the WHO Framework Convention on Tobacco Control), but a non-legal, advocacy-based approach with diet and physical activity (the Global Strategy on Diet, Physical Activity and Health). The paper assesses the merits of the Millennium Development Goals (MDGs) and the FCTC as distinct global processes for advancing health development, before considering what lessons might be learned for enhancing the implementation of the Global Strategy on Diet. While global partnerships, economic incentives, and international legal instruments could each contribute to a more effective global response to chronic diseases, the paper makes a special case for the development of international legal standards in select areas of diet and nutrition, as a strategy for ensuring that the health of future generations does not become dependent on corporate charity and voluntary commitments. A broader frame of reference for lifestyle-related chronic diseases is needed: one that draws together WHO's work in tobacco, nutrition and physical activity, and that envisages selective use of international legal obligations, non-binding recommendations, advocacy and policy advice as tools of choice for promoting different elements of the strategy.


The postdisaster prevalence of major depression relative to PTSD in survivors of the 9/11 attacks on the World Trade Center selected from affected workplaces.
Studies of survivors of the September 11, 2001 attacks on the World Trade Center in New York City suggest that postdisaster depressive disorders may be at least as prevalent, or even more prevalent, than posttraumatic stress disorder (PTSD), unlike findings from most other disaster studies. The relative prevalence and incidence of major depressive disorder (MDD) and PTSD were examined after the 9/11 attacks relative to trauma exposures. This study used full diagnostic assessment methods and careful categorization of exposure groups based on DSM-IV-TR criteria for PTSD to examine 373 employees of 9/11-affected New York City workplaces. Postdisaster new MDD episode (26%) in the entire sample was significantly more prevalent (p<.001) than 9/11-related PTSD (14%). Limiting the comparison to participants with 9/11 trauma exposures, the prevalence of postdisaster new MDD episode and 9/11-related PTSD did not differ (p=.446). The only 9/11 trauma exposure group with a significant difference in relative prevalence of MDD and PTSD were those with a 9/11 trauma-exposed close associate, for whom postdisaster new MDD episode (45%) was more prevalent (p=.046) than 9/11-related PTSD (31%). Because of the conditional definition of PTSD requiring trauma exposure that is not part of MDD criteria, prevalence comparisons of these two disorders must be limited to groups with qualifying trauma exposures to be meaningful. Findings from this study suggest distinct mechanisms underlying these two disorders that differentially relate to direct exposure to trauma vs. the magnitude of the disaster and personal connectedness to disaster and community-wide effects.


Aflatoxin regulations and global pistachio trade: insights from social network analysis.
Aflatoxins, carcinogenic toxins produced by Aspergillus fungi, contaminate maize, peanuts, and tree nuts in many regions of the world. Pistachios are the main source of human dietary aflatoxins from tree nuts worldwide. Over 120 countries have regulations for maximum allowable aflatoxin levels in food commodities. We developed social network models to analyze the association between nations' aflatoxin regulations and global trade patterns of pistachios from 1996-2010. The main pistachio producing countries are Iran and the United States (US), which together contribute to nearly 75% of the total global pistachio market. Over this time period, during which many nations developed or changed their aflatoxin regulations in pistachios, global pistachio trade patterns changed; with the US increasingly exporting to countries with stricter aflatoxin standards. The US pistachio crop has had consistently lower levels of aflatoxin than the Iranian crop over this same time period. As similar trading patterns have also been documented in maize, public health may be affected if countries without aflatoxin regulations, or with more relaxed regulations, continually import crops with higher aflatoxin contamination. Unlike the previous studies on maize, this analysis includes a dynamic element, examining how trade patterns change over time with introduction or adjustment of aflatoxin regulations.


Electron microscopy of the nuclear membrane of Amoeba proteus.
An electron microscope study of the nuclear membrane of Amoeba proteus by thin sectioning techniques has revealed an ultrastructure in the outer layer of the membrane that is homologous to the pores and annuli observed in the nuclear membranes of many other cell types studied by these techniques. An inner honeycombed layer apparently unique to Amoeba proteus is also described.


Angiostatic treatment prior to chemo- or photodynamic therapy improves anti-tumor efficacy.
Tumor vasculature is known to be poorly organized leading to increased leakage of molecules to the extravascular space. This process can potentially increase interstitial fluid pressure impairing intra-tumoral blood flow and oxygen supply, and can affect drug uptake. Anti-angiogenic therapies are believed to reduce vascular permeability, potentially reducing interstitial fluid pressure and improving the extravasation of small molecule-based chemotherapeutics. Here we show that pretreatment of human ovarian carcinoma tumors with sub-optimal doses of the VEGFR targeting tyrosine kinase inhibitor axitinib, but not the EGFR targeting kinase inhibitor erlotinib, induces a transient period of increased tumor oxygenation. Doxorubicin administered within this window was found to enter the extravascular tumor space more rapidly compared to doxorubicin when applied alone or outside this time window. Treatment with the chemotherapeutics, doxorubicin and RAPTA-C, as well as applying photodynamic therapy during this period of elevated oxygenation led to enhanced tumor growth inhibition. Improvement of therapy was not observed when applied outside the window of increased oxygenation. Taken together, these findings further confirm the hypothesis of angiostasis-induced vascular normalization and also help to understand the interactions between anti-angiogenesis and other anti-cancer strategies.


Neural organization of spoken language revealed by lesion-symptom mapping.
Studies of patients with acquired cognitive deficits following brain damage and studies using contemporary neuroimaging techniques form two distinct streams of research on the neural basis of cognition. In this study, we combine high-quality structural neuroimaging analysis techniques and extensive behavioural assessment of patients with persistent acquired language deficits to study the neural basis of language. Our results reveal two major divisions within the language system-meaning versus form and recognition versus production-and their instantiation in the brain. Phonological form deficits are associated with lesions in peri-Sylvian regions, whereas semantic production and recognition deficits are associated with damage to the left anterior temporal lobe and white matter connectivity with frontal cortex, respectively. These findings provide a novel synthesis of traditional and contemporary views of the cognitive and neural architecture of language processing, emphasizing dual routes for speech processing and convergence of white matter tracts for semantic control and/or integration.


Idiosyncratic adverse reactions to antiepileptic drugs.
Idiosyncratic drug reactions may be defined as adverse effects that cannot be explained by the known mechanisms of action of the offending agent, do not occur at any dose in most patients, and develop mostly unpredictably in susceptible individuals only. These reactions are generally thought to account for up to 10% of all adverse drug reactions, but their frequency may be higher depending on the definition adopted. Idiosyncratic reactions are a major source of concern because they encompass most life-threatening effects of antiepileptic drugs (AEDs), as well as many other reactions requiring discontinuation of treatment. Based on the underlying mechanisms, idiosyncratic reactions can be differentiated into (1) immune-mediated hypersensitivity reactions, which may range from benign skin rashes to serious conditions such as drug-related rash with eosinophilia and systemic symptoms; (2) reactions involving unusual nonimmune-mediated individual susceptibility, often related to abnormal production or defective detoxification of reactive cytotoxic metabolites (as in valproate-induced liver toxicity); and (3) off-target pharmacology, whereby a drug interacts directly with a system other than that for which it is intended, an example being some types of AED-induced dyskinesias. Although no AED is free from the potential of inducing idiosyncratic reactions, the magnitude of risk and the most common manifestations vary from one drug to another, a consideration that impacts on treatment choices. Serious consequences of idiosyncratic reactions can be minimized by knowledge of risk factors, avoidance of specific AEDs in subpopulations at risk, cautious dose titration, and careful monitoring of clinical response.


Functional sorbents for selective capture of plutonium, americium, uranium, and thorium in blood.
Self-assembled monolayer on mesoporous supports (SAMMS) are hybrid materials created from attachment of organic moieties onto very high surface area mesoporous silica. SAMMS with surface chemistries including three isomers of hydroxypyridinone, diphosphonic acid, acetamide phosphonic acid, glycinyl urea, and diethylenetriamine pentaacetate (DTPA) analog were evaluated for chelation of actinides ((239)Pu, (241)Am, uranium, thorium) from blood. Direct blood decorporation using sorbents does not have the toxicity or renal challenges associated with traditional chelation therapy and may have potential applications for critical exposure cases, reduction of nonspecific dose during actinide radiotherapy, and for sorbent hemoperfusion in renal insufficient patients, whose kidneys clear radionuclides at a very slow rate. Sorption affinity (K(d)), sorption rate, selectivity, and stability of SAMMS were measured in batch contact experiments. An isomer of hydroxypyridinone (3,4-HOPO) on SAMMS demonstrated the highest affinity for all four actinides from blood and plasma and greatly outperformed the DTPA analog on SAMMS and commercial resins. In batch contact, a fifty percent reduction of actinides in blood was achieved within minutes, and there was no evidence of protein fouling or material leaching in blood after 24 h. The engineered form of SAMMS (bead format) was further evaluated in a 100-fold scaled-down hemoperfusion device and showed no blood clotting after 2 h. A 0.2 g quantity of SAMMS could reduce 50 wt.% of 100 ppb uranium in 50 mL of plasma in 18 min and that of 500 dpm mL(-1) in 24 min. 3,4-HOPO-SAMMS has a long shelf-life in air and at room temperature for at least 8 y, indicating its feasibility for stockpiling in preparedness for an emergency. The excellent efficacy and stability of SAMMS materials in complex biological matrices suggest that SAMMS can also be used as orally administered drugs and for wound decontamination. By changing the organic groups of SAMMS, they can be used not only for actinides but also for other radionuclides. By using the mixture of these SAMMS materials, broad spectrum decorporation of radionuclides is very feasible.


An X-ray diffraction study of frog skeletal muscle during shortening near the maximum velocity.
Using an imaging plate exchanger and a synchrotron X-ray source, X-ray diffraction patterns were recorded from frog skeletal muscles shortening near the maximum velocity from a sarcomere length of 2.6 to 2.1 microns. The major findings were as follows: (1) the intensity of the second-order myosin meridional reflection, which decreased to 18% of the resting value during isometric contraction, increased to 34% during shortening, showing that the perturbation in the axial arrangement of myosin heads recovered. (2) The intensity of the third-order myosin meridional reflection increased by 32% on isometric contraction and decreased to 51% of the resting intensity during rapid shortening, showing disorder in the axial rearrangement of the myosin heads. (3) The axial Bragg spacing of the third-order meridional reflection, which increased by 1.22% during isometric contraction, was still 0.47% larger than the resting value during shortening, indicating that the increase is not entirely due to extension of the filament by tension. (4) The intensity ratio of the (1,0) and (1,1) equatorial reflections increased a small extent (from 0.57 to 0.62) during shortening, while the 5.9 nm and 5.1 nm actin layer-lines showed large intensity decreases (126 to 101% and 190 to 124% of the resting intensity, respectively), suggesting that considerable numbers of myosin heads are in the vicinity of the thin filament but not tightly bound to actin molecules during rapid shortening. These results are consistent with a model of muscle contraction in which myosin heads attach to actin in two different manners with weak and strong affinities and show that, during shortening, the number of tightly attached heads decreases by half, while the total number of attached heads decreases only slightly.


Inhibition of DNA repair protein RAD51 affects porcine preimplantation embryo development.
Homologous recombination (HR) plays a critical role in facilitating replication fork progression when the polymerase complex encounters a blocking DNA lesion, and it also serves as the primary mechanism for error-free DNA repair of double-stranded breaks. DNA repair protein RAD51 homolog 1 (RAD51) plays a central role in HR. However, the role of RAD51 during porcine early embryo development is unknown. In the present study, we examined whether RAD51 is involved in the regulation of early embryonic development of porcine parthenotes. We found that inhibition of RAD51 delayed cleavage and ceased development before the blastocyst stage. Disrupting RAD51 activity with RNAi or an inhibitor induces sustained DNA damage, as demonstrated by the formation of distinct γH2AX foci in nuclei of four-cell embryos. Inhibiting RAD51 triggers a DNA damage checkpoint by activating the ataxia telangiectasia mutated (ATM)-p53-p21 pathway. Furthermore, RAD51 inhibition caused apoptosis, reactive oxygen species accumulation, abnormal mitochondrial distribution and decreased pluripotent gene expression in blastocysts. Thus, our results indicate that RAD51 is required for proper porcine parthenogenetic activation (PA) embryo development.


Processing of emotional words measured simultaneously with steady-state visually evoked potentials and near-infrared diffusing-wave spectroscopy.
Emotional stimuli are preferentially processed compared to neutral ones. Measuring the magnetic resonance blood-oxygen level dependent (BOLD) response or EEG event-related potentials, this has also been demonstrated for emotional versus neutral words. However, it is currently unclear whether emotion effects in word processing can also be detected with other measures such as EEG steady-state visual evoked potentials (SSVEPs) or optical brain imaging techniques. In the present study, we simultaneously performed SSVEP measurements and near-infrared diffusing-wave spectroscopy (DWS), a new optical technique for the non-invasive measurement of brain function, to measure brain responses to neutral, pleasant, and unpleasant nouns flickering at a frequency of 7.5 Hz. The power of the SSVEP signal was significantly modulated by the words' emotional content at occipital electrodes, showing reduced SSVEP power during stimulation with pleasant compared to neutral nouns. By contrast, the DWS signal measured over the visual cortex showed significant differences between stimulation with flickering words and baseline periods, but no modulation in response to the words' emotional significance. This study is the first investigation of brain responses to emotional words using simultaneous measurements of SSVEPs and DWS. Emotional modulation of word processing was detected with EEG SSVEPs, but not by DWS. SSVEP power for emotional, specifically pleasant, compared to neutral words was reduced, which contrasts with previous results obtained when presenting emotional pictures. This appears to reflect processing differences between symbolic and pictorial emotional stimuli. While pictures prompt sustained perceptual processing, decoding the significance of emotional words requires more internal associative processing. Reasons for an absence of emotion effects in the DWS signal are discussed.


miR-30a inhibits endothelin A receptor and chemoresistance in ovarian carcinoma.
Drug resistance remains the major clinical barrier to successful treatment in epithelial ovarian carcinoma (EOC) patients, and the evidence of microRNA involvement in drug resistance has been recently emerging. Endothelin-1 (ET-1)/ETA receptor (ETAR) axis is aberrantly activated in chemoresistant EOC cells and elicits pleiotropic effects promoting epithelial-to-mesenchymal transition (EMT) and the acquisition of chemoresistance. However, the relationship between ETAR and miRNA is still unknown. Hence, in this study we evaluated whether dysregulation of miRNA might enhance ETAR expression in sensitive and resistant EOC cells. Based on bioinformatic tools, we selected putative miRNA able to recognize the 3'UTR of ETAR. An inverse correlation was observed between the expression levels of miR-30a and ETAR in both EOC cell lines and tumor samples. miR-30a was found to specifically bind to the 3'UTR of ETAR mRNA, indicating that ETAR is a direct target of miR-30a. Overexpression of miR-30a decreased Akt and mitogen activated protein kinase signaling pathway activation, cell proliferation, invasion, plasticity, EMT marker levels, and vascular endothelial growth factor release. Interestingly, ectopic expression of miR-30a re-sensitized platinum-resistant EOC cells to cisplatinum-induced apoptosis. Consistently, resistant EOC xenografts overexpressing miR-30a resulted in significantly less tumor growth than controls. Together our study provides a new perspective on the regulatory mechanism of ETAR gene. Interestingly, our findings highlight that blockade of ETAR regulatory axis is the mechanism underlying the tumor suppressor function of miR-30a in chemoresistant EOC cells.


Coccidioidomycosis in alpacas in the southwestern United States.
An anonymous web-based survey of alpaca owners was used to learn more about the clinical presentation, diagnosis, and treatment of coccidioidomycosis in alpacas in the United States. Thirty-seven owners, with 1,117 alpacas, completed the survey. Over 4% of alpacas included in the study were diagnosed with coccidioidomycosis between 2005 and 2016 (5 post mortem, 46 clinically). Immunodiffusion titers ranged from 1:4 to ≥1:256 in sick animals. Alpacas residing in Arizona counties with a high incidence of human disease were 5.8 times more likely to contract coccidioidomycosis than animals residing in other areas of the state. Treatment was reported in 23 alpacas, and 78% of those animals died or were euthanized. Necropsy records from a veterinary diagnostic laboratory in Tucson, AZ were reviewed to estimate the severity of disease in this species. Nine cases identified for review died of disseminated coccidioidomycosis; the disease was extensive in most animals, with the lungs, lymph nodes, and liver the most frequently affected. Alpacas appear to be highly susceptible to severe illness as a result of infection by Coccidioides spp., frequently resulting in death. More research is needed to better understand the epidemiology, clinical signs, and treatment protocols for coccidioidomycosis in alpacas.


Infertility.
Infertility is defined as failure to achieve pregnancy during one year of frequent, unprotected intercourse. Evaluation generally begins after 12 months, but it can be initiated earlier if infertility is suspected based on history or if the female partner is older than 35 years. Major causes of infertility include male factors, ovarian dysfunction, tubal disease, endometriosis, and uterine or cervical factors. A careful history and physical examination of each partner can suggest a single or multifactorial etiology and can direct further investigation. Ovulation can be documented with a home urinary luteinizing hormone kit. Hysterosalpingography and pelvic ultrasonography can be used to screen for uterine and fallopian tube disease. Hysteroscopy and/or laparoscopy can be used if no abnormalities are found on initial screening. Women older than 35 years also may benefit from ovarian reserve testing of follicle-stimulating hormone and estradiol levels on day 3 of the menstrual cycle, the clomiphene citrate challenge test, or pelvic ultrasonography for antral follicle count to determine treatment options and the likelihood of success. Options for the treatment of male factor infertility include gonadotropin therapy, intrauterine insemination, or in vitro fertilization. Infertility attributed to ovulatory dysfunction often can be treated with oral ovulation-inducing agents in a primary care setting. Women with poor ovarian reserve have more success with oocyte donation. In certain cases, tubal disease may be treatable by surgical repair or by in vitro fertilization. Infertility attributed to endometriosis may be amenable to surgery, induction of ovulation with intrauterine insemination, or in vitro fertilization. Unexplained infertility may be managed with ovulation induction, intrauterine insemination, or both. The overall likelihood of successful pregnancy with treatment is nearly 50 percent.


Dietary Inulin and Trichuris suis Infection Promote Beneficial Bacteria Throughout the Porcine Gut.
The gut microbiota (GM) displays a profound ability to adapt to extrinsic factors, such as gastrointestinal pathogens and/or dietary alterations. Parasitic worms (helminths) and host-associated GM share a long co-evolutionary relationship, exerting mutually modulatory effects which may impact the health of the host. Moreover, dietary components such as prebiotic fibers (e.g. inulin) are capable of modulating microbiota toward a composition often associated with a healthier gut function. The effect of helminth infection on the host microbiota is still equivocal, and it is also unclear how parasites and prebiotic dietary components interact to influence the microbiota and host health status. Some helminths, such as Trichuris suis (porcine whipworm), also exhibit strong immunomodulatory and anti-inflammatory effects. We therefore explored the effects of T. suis, alone and in interaction with inulin, both in fecal microbiota during the infection period and luminal microbiota across four intestinal segments at the end of a 4-week infection period. We observed that T. suis generally had minimal, but mainly positive, effects on the microbiota. T. suis increased the relative abundance of bacterial genera putatively associated with gut health such as Prevotella, and decreased bacteria such as Proteobacteria that have been associated with dysbiosis. Interestingly, dietary inulin interacted with T. suis to enhance these effects, thereby modulating the microbiota toward a composition associated with reduced inflammation. Our results show that administration of T. suis together with the consumption of prebiotic inulin may have the potential to positively affect gut health.