Timing of increased autistic disorder cumulative incidence.
Autistic disorder (AD) is a severe neurodevelopmental disorder typically identified in early childhood. Both genetic and environmental factors are implicated in its etiology. The number of individuals identified as having autism has increased dramatically in recent years, but whether some proportion of this increase is real is unknown. If real, susceptible populations may have exposure to controllable exogenous stressors. Using literature AD data from long-term (approximately 10-year) studies, we determined cumulative incidence of AD for each cohort within each study. These data for each study were examined for a changepoint year in which the AD cumulative incidence first increased. We used data sets from Denmark, California, Japan, and a worldwide composite of studies. In the Danish, California, and worldwide data sets, we found that an increase in AD cumulative incidence began about 1988-1989. The Japanese study (1988-1996) had AD cumulative incidence increasing continuously, and no changepoint year could be calculated. Although the debate about the nature of increasing autism continues, the potential for this increase to be real and involve exogenous environmental stressors exists. The timing of an increase in autism incidence may help in screening for potential candidate environmental stressors.


Biophysics (and sociology) of ceramides.
In the past decade, the long-neglected ceramides (N-acylsphingosines) have become one of the most attractive lipid molecules in molecular cell biology, because of their involvement in essential structures (stratum corneum) and processes (cell signalling). Most natural ceramides have a long (16-24 C atoms) N-acyl chain, but short N-acyl chain ceramides (two to six C atoms) also exist in Nature, apart from being extensively used in experimentation, because they can be dispersed easily in water. Long-chain ceramides are among the most hydrophobic molecules in Nature, they are totally insoluble in water and they hardly mix with phospholipids in membranes, giving rise to ceramide-enriched domains. In situ enzymic generation, or external addition, of long-chain ceramides in membranes has at least three important effects: (i) the lipid monolayer tendency to adopt a negative curvature, e.g. through a transition to an inverted hexagonal structure, is increased, (ii) bilayer permeability to aqueous solutes is notoriously enhanced, and (iii) transbilayer (flip-flop) lipid motion is promoted. Short-chain ceramides mix much better with phospholipids, promote a positive curvature in lipid monolayers, and their capacities to increase bilayer permeability or transbilayer motion are very low or non-existent.


Pregabalin in treatment-refractory fibromyalgia.
Fibromyalgia is a chronic musculoskeletal pain disorder. The pain can be intractable and may not respond to commonly-used treatments, such as tricyclic antidepressants and opioids. To evaluate pregabalin response in the subset of patients with fibromyalgia whose pain had been judged refractory to other treatments. Patients had previously participated in a controlled trial of pregabalin and had moderate to severe pain despite treatment with gabapentin, a tricyclic antidepressant, and a third medication (e.g., other anticonvulsants, opioid, selective serotonin reuptake inhibitors, tramadol). Flexible-dose pregabalin 150-600 mg/day was added for 3-month treatment cycles, each followed by 3- to 28-day pregabalin "drug holiday" that lasted until a relapse occurred. Pain intensity was measured using the visual analogue scale of the Short-Form McGill Pain Questionnaire completed at baseline, the end of each 3-month treatment period and at the relapse visit. Analysis was at 15 months (after 5 cycles). In total, 25 patients were included and 19 completed the 15-month analysis period. At baseline, 88% were receiving ≥1 pain medication. Pregabalin 150-600 mg/day was associated with statistically significant, clinically meaningful pain reduction during each treatment cycle. Pain quickly returned to baseline levels during the "drug holidays" in a median time of 2-4 days. Somnolence (n=5) and dizziness (n=4) were the most common adverse events. These results suggest that pregabalin may be beneficial in patients with fibromyalgia who have had an unsatisfactory response to treatment with other medications.


Hybrid phenolic-inducible promoters towards construction of self-inducible systems for microbial lignin valorization.
Engineering strategies to create promoters that are both higher strength and tunable in the presence of inexpensive compounds are of high importance to develop metabolic engineering technologies that can be commercialized. Lignocellulosic biomass stands out as the most abundant renewable feedstock for the production of biofuels and chemicals. However, lignin a major polymeric component of the biomass is made up of aromatic units and remains as an untapped resource. Novel synthetic biology tools for the expression of heterologous proteins are critical for the effective engineering of a microbe to valorize lignin. This study demonstrates the first successful attempt in the creation of engineered promoters that can be induced by aromatics present in lignocellulosic hydrolysates to increase heterologous protein production. A hybrid promoter engineering approach was utilized for the construction of phenolic-inducible promoters of higher strength. The hybrid promoters were constructed by replacing the spacer region of an endogenous promoter, PemrR present in E. coli that was naturally inducible by phenolics. In the presence of vanillin, the engineered promoters Pvtac, Pvtrc, and Pvtic increased protein expression by 4.6-, 3.0-, and 1.5-fold, respectively, in comparison with a native promoter, PemrR. In the presence of vanillic acid, Pvtac, Pvtrc, and Pvtic improved protein expression by 9.5-, 6.8-, and 2.1-fold, respectively, in comparison with PemrR. Among the cells induced with vanillin, the emergence of a sub-population constituting the healthy and dividing cells using flow cytometry was observed. The analysis also revealed this smaller sub-population to be the primary contributor for the increased expression that was observed with the engineered promoters. This study demonstrates the first successful attempt in the creation of engineered promoters that can be induced by aromatics to increase heterologous protein production. Employing promoters inducible by phenolics will provide the following advantages: (1) develop substrate inducible systems; (2) lower operating costs by replacing expensive IPTG currently used for induction; (3) develop dynamic regulatory systems; and (4) provide flexibility in operating conditions. The flow cytometry findings strongly suggest the need for novel approaches to maintain a healthy cell population in the presence of phenolics to achieve increased heterologous protein expression and, thereby, valorize lignin efficiently.


Nanostructured hydrotreating catalysts for electrochemical hydrogen evolution.
Progress in catalysis is driven by society's needs. The development of new electrocatalysts to make renewable and clean fuels from abundant and easily accessible resources is among the most challenging and demanding tasks for today's scientists and engineers. The electrochemical splitting of water into hydrogen and oxygen has been known for over 200 years, but in the last decade and motivated by the perspective of solar hydrogen production, new catalysts made of earth-abundant materials have emerged. Here we present an overview of recent developments in the non-noble metal catalysts for electrochemical hydrogen evolution reaction (HER). Emphasis is given to the nanostructuring of industrially relevant hydrotreating catalysts as potential HER electrocatalysts. The new syntheses and nanostructuring approaches might pave the way for future development of highly efficient catalysts for energy conversion.


Loss of STEP61 couples disinhibition to N-methyl-d-aspartate receptor potentiation in rodent and human spinal pain processing.
Dysregulated excitability within the spinal dorsal horn is a critical mediator of chronic pain. In the rodent nerve injury model of neuropathic pain, BDNF-mediated loss of inhibition (disinhibition) gates the potentiation of excitatory GluN2B N-methyl-d-aspartate receptor (NMDAR) responses at lamina I dorsal horn synapses. However, the centrality of this mechanism across pain states and species, as well as the molecular linker involved, remain unknown. Here, we show that KCC2-dependent disinhibition is coupled to increased GluN2B-mediated synaptic NMDAR responses in a rodent model of inflammatory pain, with an associated downregulation of the tyrosine phosphatase STEP61. The decreased activity of STEP61 is both necessary and sufficient to prime subsequent phosphorylation and potentiation of GluN2B NMDAR by BDNF at lamina I synapses. Blocking disinhibition reversed the downregulation of STEP61 as well as inflammation-mediated behavioural hypersensitivity. For the first time, we characterize GluN2B-mediated NMDAR responses at human lamina I synapses and show that a human ex vivo BDNF model of pathological pain processing downregulates KCC2 and STEP61 and upregulates phosphorylated GluN2B at dorsal horn synapses. Our results demonstrate that STEP61 is the molecular brake that is lost following KCC2-dependent disinhibition and that the decrease in STEP61 activity drives the potentiation of excitatory GluN2B NMDAR responses in rodent and human models of pathological pain. The ex vivo human BDNF model may thus form a translational bridge between rodents and humans for identification and validation of novel molecular pain targets.


Acquired type 3-like von Willebrand syndrome preceded full-blown systemic lupus erythematosus.
We report a quite rare case of acquired type 3-like von Willebrand syndrome (vWS) that preceded full-blown systemic lupus erythematosus (SLE). A 16-year-old woman with no previous disease history and no family history of hemorrhagic diathesis was referred to our hospital because of recurrent epistaxis and gingival bleeding. She was diagnosed as having atypical type 3 von Willebrand disease because of prolonged bleeding time with normal platelet count and prolonged activated partial thromboplastin time (aPTT), and an almost complete absence of von Willebrand factor (vWF) antigen, ristocetin cofactor activity (vWF:RCo) and ristocetin-induced platelet agglutination (RIPA). Furthermore, electrophoretic analysis of plasma vWF revealed a trace amount of vWF and an absence of the multimeric form of vWF. Infusions of either vasopressin or factor VIII/vWF concentrates improved bleeding symptoms and corrected the aPTT and RIPA. However, she complained of low-grade fever, general fatigue and polyarthralgia 5 months later, and leukocytepenia and hypo-complementemia developed. Anti-double-stranded DNA antibodies and lupus erythematosus cells became positive. These findings were compatible with SLE. Mixing the patient's platelet-poor plasma (PPP) with normal platelet-rich plasma (PRP) (PPP/PRP = 2/1) resulted in a complete inhibition of RIPA, suggesting the presence of vWF inhibitor in her plasma. Treatment with prednisolone (40 mg/day) started and the bleeding tendency gradually improved. One month later, all of the laboratory data including aPTT, bleeding time, RIPA and vWF:RCo became normal. These findings indicate that she has an acquired type 3-like vWS associated with SLE.


Brain size affects responsiveness in mating behaviour to variation in predation pressure and sex ratio.
Despite ongoing advances in sexual selection theory, the evolution of mating decisions remains enigmatic. Cognitive processes often require simultaneous processing of multiple sources of information from environmental and social cues. However, little experimental data exist on how cognitive ability affects such fitness-associated aspects of behaviour. Using advanced tracking techniques, we studied mating behaviours of guppies artificially selected for divergence in relative brain size, with known differences in cognitive ability, when predation threat and sex ratio was varied. In females, we found a general increase in copulation behaviour in when the sex ratio was female biased, but only large-brained females responded with greater willingness to copulate under a low predation threat. In males, we found that small-brained individuals courted more intensively and displayed more aggressive behaviours than large-brained individuals. However, there were no differences in female response to males with different brain size. These results provide further evidence of a role for female brain size in optimal decision-making in a mating context. In addition, our results indicate that brain size may affect mating display skill in male guppies. We suggest that it is important to consider the association between brain size, cognitive ability and sexual behaviour when studying how morphological and behavioural traits evolve in wild populations.


PilY1 Promotes Legionella pneumophila Infection of Human Lung Tissue Explants and Contributes to Bacterial Adhesion, Host Cell Invasion, and Twitching Motility.
Legionnaires' disease is an acute fibrinopurulent pneumonia. During infection Legionella pneumophila adheres to the alveolar lining and replicates intracellularly within recruited macrophages. Here we provide a sequence and domain composition analysis of the L. pneumophila PilY1 protein, which has a high homology to PilY1 of Pseudomonas aeruginosa. PilY1 proteins of both pathogens contain a von Willebrand factor A (vWFa) and a C-terminal PilY domain. Using cellular fractionation, we assigned the L. pneumophila PilY1 as an outer membrane protein that is only expressed during the transmissive stationary growth phase. PilY1 contributes to infection of human lung tissue explants (HLTEs). A detailed analysis using THP-1 macrophages and A549 lung epithelial cells revealed that this contribution is due to multiple effects depending on host cell type. Deletion of PilY1 resulted in a lower replication rate in THP-1 macrophages but not in A549 cells. Further on, adhesion to THP-1 macrophages and A549 epithelial cells was decreased. Additionally, the invasion into non-phagocytic A549 epithelial cells was drastically reduced when PilY1 was absent. Complementation variants of a PilY1-negative mutant revealed that the C-terminal PilY domain is essential for restoring the wild type phenotype in adhesion, while the putatively mechanosensitive vWFa domain facilitates invasion into non-phagocytic cells. Since PilY1 also promotes twitching motility of L. pneumophila, we discuss the putative contribution of this newly described virulence factor for bacterial dissemination within infected lung tissue.


Pegfilgrastim support for full delivery of BEACOPP-14 chemotherapy for patients with high-risk Hodgkin's lymphoma: results of a phase II study.
The primary endpoint of this feasibility study was to determine whether pegfilgrastim support could enable the delivery of the full dose of BEACOPP chemotherapy every 14 days on schedule. Forty-one patients with high-risk Hodgkin's lymphoma were randomized to receive pegfilgrastim (6 mg) on day 4 or 8 of each cycle. Eighty-one percent of cycles administered were delivered at full dose and on schedule (FDOS). Response was retrospectively assessed in 27 patients at 6 months; 23 of these 27 patients (85%) achieved a complete response and one (4%) achieved a partial response. Toxicities were mostly moderate in intensity. These results support the feasibility of delivering full dose, on schedule BEACOPP-14, chemotherapy with pegfilgrastim support.


Addressing the escalating burden of chronic diseases in India: Need for strengthening primary care.
The growing epidemic of noncommunicable diseases (NCDs) has impacted the national health systems, policies, and socioeconomic developments, thereby leading to increasing country level disparities. Despite substantial improvements in health indicators made in the past decade, the Indian health-care system continues to contribute disproportionately to the global disease burden, wherein NCDs holds significant prominence. Against this background, the present review analyzes the current NCD landscape from the perspective of India's health system preparedness toward meeting this growing challenge. Implementation and delivery of strategies and interventions are often impeded by existing grass root level challenges. Recognizing the importance of effective primary care, the review highlights the importance of implementing affordable, accessible, and comprehensive interventions, and delivering them at societal, a community and individual level. This simultaneously calls for strengthening of the primary care system through appropriate strategy and policy frameworks. Toward addressing India-specific needs in NCD prevention and management, concerted efforts on development of robust surveillance mechanisms, intersectoral and interdepartmental collaborations, integration of national programs, enhanced role of education and awareness should be made, to ensure effectivity, scale-up, and outreach of services in primary care.


Adaptive optoelectronic camouflage systems with designs inspired by cephalopod skins.
Octopus, squid, cuttlefish, and other cephalopods exhibit exceptional capabilities for visually adapting to or differentiating from the coloration and texture of their surroundings, for the purpose of concealment, communication, predation, and reproduction. Long-standing interest in and emerging understanding of the underlying ultrastructure, physiological control, and photonic interactions has recently led to efforts in the construction of artificial systems that have key attributes found in the skins of these organisms. Despite several promising options in active materials for mimicking biological color tuning, existing routes to integrated systems do not include critical capabilities in distributed sensing and actuation. Research described here represents progress in this direction, demonstrated through the construction, experimental study, and computational modeling of materials, device elements, and integration schemes for cephalopod-inspired flexible sheets that can autonomously sense and adapt to the coloration of their surroundings. These systems combine high-performance, multiplexed arrays of actuators and photodetectors in laminated, multilayer configurations on flexible substrates, with overlaid arrangements of pixelated, color-changing elements. The concepts provide realistic routes to thin sheets that can be conformally wrapped onto solid objects to modulate their visual appearance, with potential relevance to consumer, industrial, and military applications.


Poly (lactic-co-glycolic acid)/graphene oxide composites combined with electrical stimulation in wound healing: preparation and characterization.
In this study, we fabricated multifunctional, electrically conductive composites by incorporating graphene oxide (GO) into a poly (lactic-co-glycolic acid) (PLGA) copolymer for wound repair. Furthermore, the resultant composites were coupled with electrical stimulation to further improve the therapeutic effect of wound repair. We evaluated the surface morphology of the composites, as well as their physical properties, cytotoxicity, and antibacterial activity, along with the combined effects of composites and electrical stimulation (ES) in a rat model of wound healing. Application of the PLGA/GO composites to full-thickness wounds confirmed their advantageous biological properties, as evident from the observed improvements in wound-specific mechanical properties, biocompatibility, and antibacterial activity. Additionally, we found that the combination of composites and ES improved composite-mediated cell survival and accelerated wound healing in vivo by promoting neovascularization and the formation of type I collagen. These results demonstrated that combined treatment with the PLGA/GO composite and ES promoted vascularization and epidermal remodeling and accelerated wound healing in rats, thereby suggesting the efficacy of PLGA/GO+ES for broad applications associated with wound repair.


Monomeric recombinant TCR ligand reduces relapse rate and severity of experimental autoimmune encephalomyelitis in SJL/J mice through cytokine switch.
Our previous studies demonstrated that oligomeric recombinant TCR ligands (RTL) can treat clinical signs of experimental autoimmune encephalomyelitis (EAE) and induce long-term T cell tolerance against encephalitogenic peptides. In the current study, we produced a monomeric I-A(s)/PLP 139-151 peptide construct (RTL401) suitable for use in SJL/J mice that develop relapsing disease after injection of PLP 139-151 peptide in CFA. RTL401 given i.v. or s.c. but not empty RTL400 or free PLP 139-151 peptide prevented relapses and significantly reduced clinical severity of EAE induced by PLP 139-151 peptide in SJL/J or (C57BL/6 x SJL)F(1) mice, but did not inhibit EAE induced by PLP 178-191 or MBP 84-104 peptides in SJL/J mice, or MOG 35-55 peptide in (C57BL/6 x SJL/J)F(1) mice. RTL treatment of EAE caused stable or enhanced T cell proliferation and secretion of IL-10 in the periphery, but reduced secretion of inflammatory cytokines and chemokines. In CNS, there was a modest reduction of inflammatory cells, reduced expression of very late activation Ag-4, lymphocyte function-associated Ag-1, and inflammatory cytokines, chemokines, and chemokine receptors, but enhanced expression of Th2-related factors, IL-10, TGF-beta3, and CCR3. These results suggest that monomeric RTL therapy induces a cytokine switch that curbs the encephalitogenic potential of PLP 139-151-specific T cells without fully preventing their entry into CNS, wherein they reduce the severity of inflammation. This mechanism differs from that observed using oligomeric RTL therapy in other EAE models. These results strongly support the clinical application of this novel class of peptide/MHC class II constructs in patients with multiple sclerosis who have focused T cell responses to known encephalitogenic myelin peptides.


Integrative transcriptomic analysis of tissue-specific metabolic crosstalk after myocardial infarction.
Myocardial infarction (MI) promotes a range of systemic effects, many of which are unknown. Here, we investigated the alterations associated with MI progression in heart and other metabolically active tissues (liver, skeletal muscle, and adipose) in a mouse model of MI (induced by ligating the left ascending coronary artery) and sham-operated mice. We performed a genome-wide transcriptomic analysis on tissue samples obtained 6- and 24 hr post MI or sham operation. By generating tissue-specific biological networks, we observed: (1) dysregulation in multiple biological processes (including immune system, mitochondrial dysfunction, fatty-acid beta-oxidation, and RNA and protein processing) across multiple tissues post MI and (2) tissue-specific dysregulation in biological processes in liver and heart post MI. Finally, we validated our findings in two independent MI cohorts. Overall, our integrative analysis highlighted both common and specific biological responses to MI across a range of metabolically active tissues.


Accountable care organization hospitals differ in health IT capabilities.
The aim of this study was to evaluate health information technology (IT) adoption in hospitals participating in accountable care organizations (ACOs) and compare this adoption to non-ACO hospitals. A cross-sectional sample of US nonfederal, acute care hospitals with data from 3 matched sources: the 2013 American Hospital Association (AHA) Annual Survey, the 2013 AHA Survey of Care Systems and Payments (CSP), and the 2014 AHA Information Technology Supplement. To compare health IT adoption in ACO- and non-ACO hospitals, we created measures of Meaningful Use (MU) Stage 1 and Stage 2 core and menu criteria, patient engagement-oriented health IT, and health information exchange (HIE) participation. Adoption was compared using both naïve and multivariate logit models. Of the 393 ACO hospitals and 810 non-ACO hospitals, a greater percentage of ACO hospitals were capable of meeting MU Stage 1 (50.9% vs 41.6%; P < .01) and Stage 2 (7.6% vs 4.8%; P < .05), having patient engagement health IT (39.8% vs 15.2%; P < .001), and participating in HIE (49.0% vs 30.1%; P < .001). In adjusted models, no difference was found between ACO and non-ACO hospital ability to meet MU Stage 1 or Stage 2, but ACO hospitals were more likely to have patient engagement health IT (odds ratio (OR), 2.20; 95% CI, 1.59-3.04) and be HIE participants (OR, 1.41; 95% CI, 1.03-1.92). ACO-participating hospitals appear to be focused more on adopting health IT that aligns with broader strategic goals rather than those that achieve MU. Aligning adoption with quality and payment reform may be a productive path forward to encourage hospital health IT adoption behavior.


A yeast FRET biosensor enlightens cAMP signaling.
The cAMP-PKA signaling cascade in budding yeast regulates adaptation to changing environments. We developed yEPAC, a FRET-based biosensor for cAMP measurements in yeast. We used this sensor with flow cytometry for high-throughput single cell-level quantification during dynamic changes in response to sudden nutrient transitions. We found that the characteristic cAMP peak differentiates between different carbon source transitions and is rather homogenous among single cells, especially for transitions to glucose. The peaks are mediated by a combination of extracellular sensing and intracellular metabolism. Moreover, the cAMP peak follows the Weber-Fechner law; its height scales with the relative, and not the absolute, change in glucose. Last, our results suggest that the cAMP peak height conveys information about prospective growth rates. In conclusion, our yEPAC-sensor makes possible new avenues for understanding yeast physiology, signaling, and metabolic adaptation.


PITX2-dependent gene regulation in atrial fibrillation and rhythm control.
Atrial fibrillation (AF) is a common arrhythmia. Better prevention and treatment of AF are needed to reduce AF-associated morbidity and mortality. There are several major mechanisms that cause AF in patients, including a genetic predisposition to develop AF. Genome-wide association studies have identified genetic variants associated with AF populations, with the strongest hits clustering on chromosome 4q25, close to the gene for the homeobox transcription factor PITX2. The effect of these common gene variants on cardiac PITX2 mRNA is currently under study. PITX2 protein regulates right-left differentiation of the embryonic heart, thorax and aorta. PITX2 is expressed in the adult left atrium, but much less so in other heart chambers. Pitx2 deficiency results in electrical and structural remodelling, and impaired repair of the heart in murine models, all of which may influence AF through divergent mechanisms. PITX2 levels and single nucleotide polymorphisms on chromosome 4q25 may also be a predictor of the effectiveness of anti-arrhythmic drug therapy.


Anthropogenic and natural influences in the evolution of lower stratospheric cooling.
Observations reveal that the substantial cooling of the global lower stratosphere over 1979-2003 occurred in two pronounced steplike transitions. These arose in the aftermath of two major volcanic eruptions, with each cooling transition being followed by a period of relatively steady temperatures. Climate model simulations indicate that the space-time structure of the observed cooling is largely attributable to the combined effect of changes in both anthropogenic factors (ozone depletion and increases in well-mixed greenhouse gases) and natural factors (solar irradiance variation and volcanic aerosols). The anthropogenic factors drove the overall cooling during the period, and the natural ones modulated the evolution of the cooling.


Preanalytical quality improvement: in quality we trust.
Total quality in laboratory medicine should be defined as the guarantee that each activity throughout the total testing process is correctly performed, providing valuable medical decision-making and effective patient care. In the past decades, a 10-fold reduction in the analytical error rate has been achieved thanks to improvements in both reliability and standardization of analytical techniques, reagents, and instrumentation. Notable advances in information technology, quality control and quality assurance methods have also assured a valuable contribution for reducing diagnostic errors. Nevertheless, several lines of evidence still suggest that most errors in laboratory diagnostics fall outside the analytical phase, and the pre- and postanalytical steps have been found to be much more vulnerable. This collective paper, which is the logical continuum of the former already published in this journal 2 years ago, provides additional contribution to risk management in the preanalytical phase and is a synopsis of the lectures of the 2nd European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)-Becton Dickinson (BD) European Conference on Preanalytical Phase meeting entitled "Preanalytical quality improvement: in quality we trust" (Zagreb, Croatia, 1-2 March 2013). The leading topics that will be discussed include quality indicators for preanalytical phase, phlebotomy practices for collection of blood gas analysis and pediatric samples, lipemia and blood collection tube interferences, preanalytical requirements of urinalysis, molecular biology hemostasis and platelet testing, as well as indications on best practices for safe blood collection. Auditing of the preanalytical phase by ISO assessors and external quality assessment for preanalytical phase are also discussed.


Antihyperglycemic Medication Use Among Medicare Beneficiaries With Heart Failure, Diabetes Mellitus, and Chronic Kidney Disease.
Diabetes mellitus, heart failure (HF), and chronic kidney disease are common comorbidities, but overall use and safety of antihyperglycemic medications (AHMs) among patients with these comorbidities are poorly understood. Using Get With the Guidelines-Heart Failure and linked Medicare Part D data, we assessed AHM use within 90 days of hospital discharge among HF patients with diabetes mellitus discharged from Get With the Guidelines-Heart Failure hospitals between January 1, 2006, and October 1, 2011. We further summarized use by renal function and assessed renal contraindicated AHM use for patients with estimated glomerular filtration rate <30 mL/min/1.73m(2). Among 8791 patients meeting inclusion criteria, the median age was 77 (interquartile range 71-83), 62.3% were female, median body mass index was 29.7 (interquartile range 25.5-35.3), median hemoglobin A1c was 6.8 (interquartile range 6.2-7.8), and 34% had ejection fraction <40%. 74.9% of patients filled a prescription for an AHM, with insulin (39.5%), sulfonylureas (32.4%), and metformin (17%) being the most commonly used AHMs. Insulin use was higher and sulfonylurea/metformin use was lower among patients with lower renal function classes. Among 1512 patients with estimated glomerular filtration rate <30 mL/min/1.73m(2), 35.4% filled prescriptions for renal contraindicated AHMs per prescribing information, though there was a trend toward lower renal contraindicated AHM use over time (Cochran-Mantel-Haenszel row-mean score test P=0.048). Although use of other AHMs was low overall, thiazolidinediones were used in 6.6% of HF patients, and dipeptidyl peptidase-4 inhibitors were used in 5.1%, with trends for decreasing thiazolidinedione use and increased dipeptidyl peptidase-4 inhibitor use over time (P<0.001). Treatment of diabetes mellitus in patients with HF and chronic kidney disease is complex, and these patients are commonly treated with renal contraindicated AHMs, including over 6% receiving a thiazolidinedione, despite known concerns regarding HF. More research regarding safety and efficacy of various AHMs among HF patients is needed.


The difficulty with audit of high grade cervical cytology in the absence of a national screening programme.
Audit is an essential requirement of a cervical screening programme to ensure that laboratories are practising to agreed standards, and to ensure high quality patient care. The aim of this study was to assess the ability to audit high-grade cytology smear reports in a large cervical cytology laboratory in Ireland, where a nationally organised screening programme does not exist. Seven hundred and five questionnaires were forwarded to smear takers requesting follow-up data regarding high grade smear results from 2003. Seventy-four percent of the questionnaires were returned containing insufficient data, with a "don't know result" rate of >50%. This attempt at detailed audit took place 5 years ago. Annual internal audit continues to the best of the laboratory's ability but the situation, in terms of a centralised database in the context of a national screening programme, remains unchanged. A National Cervical Cytology Screening Programme is essential to centralise patient data, to allow for improved patient care, patient follow-up and audit.


How much and what type: analysis of the first year of the acute care surgery operative case log.
A case log was created by the American Association for the Surgery of Trauma Acute Care Surgery (ACS) committee to track trainee operative experiences, allowing them to enter their cases in the form of Current Procedural Terminology (CPT) codes. We hypothesized that the number of cases an ACS trainee performed would be similar to the expectations of a fifth-year general surgery resident and that the current list of essential and desired cases (E/D list) would accurately reflect cases done by ACS trainees. The database was queried from July 1, 2011, to June 30, 2012. Trainees were classified as those in American Association for the Surgery of Trauma-accredited fellowships (ACC) and those in ACS fellowships not accredited (non-ACC). CPT codes were mapped to the E/D list. Cases entered manually were individually reviewed and assigned a CPT code if possible or listed as "noncodable." To compensate for nonoperative rotations and noncompliance, case numbers were analyzed both annually and monthly to estimate average case numbers for all trainees. In addition, case logs of trainees were compared with the E/D list to assess how well it reflected actual trainee experience. Eighteen ACC ACS and 11 non-ACC ACS trainees performed 16.4 (12.6) cases per month compared with 15.7 (14.2) cases for non-ACC ACS fellows (p = 0.71). When annualized, trainees performed, on average, 195 cases per year. Annual analysis led to similar results. The E/D list captured only approximately 50% of the trainees' operative experience. Only 77 cases were categorized as pediatric. ACS trainees have substantial operative experience averaging nearly 200 major cases during their ACS year. However, high variability exists in the number of essential or desirable cases being performed with approximately 50% of the fellows' operative experience falling outside the E/D list of cases. Modification of the fellows' operative experience and/or the rotation requirements seems to be needed to provide experience in E/D cases.


Gene expression, telomere and cognitive deficit analysis as a function of Chornobyl radiation dose and age: from in utero to adulthood.
Objective - to estimate the possible effects of low dose ionizing radiation on human cognitive function in adult hood and in utero. Cognitive tests, telomere length and expression of genes regulating telomere function were studied in Chornobyl cleanup workers who were exposed to doses under 500 mSv (n = 326) and subjects exposed in utero during the first days after the accident Prypiat town (n = 104). The neurocognitive assessment covered mem ory, attention, language, executive and visiospatial functions. In young adults after prenatal exposure a relation ship was analyzed between a cognitive function and radiation dose to foetus, brain and thyroid gland. Internal con trols were used for both groups - the group of Chornobyl cleanup workers exposed in doses less than 20 mSv and an age matched comparison group from radioactively contaminated areas for subjects exposed in utero. Cognitive functions in cleanup workers exposed to ionizing radiation at adulthood are characterized by symptoms of a mild cognitive impairment according to the MMSE (mean group score 25,58 ± 2,95) and a significant ly higher level of mental disorders according to the BPRS in a dose related manner. Cleanup workers exposed to doses over 500 mSv demonstrate a significant cognitive deficit in comparison with those exposed below 500 mSv and espe cially non exposed patients. Subjects exposed in utero during the check at age of 25-27 years exhibit an excess of the disorders of autonomic nervous system (ICD 10: G90). Neurological microsymptoms as well as neurotic, stress relat ed and somatoform disorders (F40-F48) dominate. Relationship were revealed between the TERT, TERF1, TERF2 genes expression, relative telomere length (RTL),cognitive deficit and cerebrovascular pathology, radiation dose and age. Telomere length in cleanup workers is sreduced after 50 years (6.1 %). The most significant reduction in telomere length is shown after 70 years (11.7 %). Negative correlation was found between telomere length and degree of cog nitive deficit (MMSE scale) and between age and degree of cognitive deficit. The RTL is significantly decreased in groups of persons with cognitive deficit compared to a comparison group. Telomere length at the late period after low dose radiation exposure is downregulated by the high TERF2 gene expression combined with low expression of TERT gene. After exposure to doses over 250-500 mSv a cognitive deficit and dementia were associated with a substantial increase in TERT gene expression, overexpression of TERF1 and decrease in expression of TERF2 gene. A relationship was revealed between the TERF2 gene expression and CD95+ cell fraction susceptible to apoptosis. This study shows that cognitive deficit in humans at a late period after radiation exposure is influ enced by dose, age at exposure and gene regulation of telomere function.


Neurofilament light chain protein in neurodegenerative dementia: A systematic review and network meta-analysis.
The diagnostic value of neurofilament light chain protein in neurodegenerative dementia diseases is still controversial. A systematic literature search was performed to identify relevant case-control studies conducted through October 2018. Traditional and net meta-analyses were performed based on 42 studies that tested the diagnostic performance of neurofilament light chain protein (NfL) concentration in CSF and serum/plasma from patients with neurodegenerative dementia. CSF and serum/plasma NfL levels were significantly increased in patients with neurodegenerative dementia diseases. Network meta-analysis showed a significant reduction in CSF NfL levels during mild cognitive impairment, whereas an increase was observed in vascular dementia compared to Alzheimer's disease. Surface under the cumulative ranking curve and cluster analysis showed that the NfL concentration in CSF (vascular dementia, frontotemporal dementia, and Alzheimer's disease) and serum/plasma (frontotemporal dementia and Alzheimer's disease) ranked first among neurodegenerative dementia diseases. NfL is an important biomarker that can help clinical neurologists make early diagnoses of neurodegenerative diseases, so patients can receive prompt treatment.


The Ca2(+)-binding glycoprotein SPARC modulates cell cycle progression in bovine aortic endothelial cells.
SPARC (secreted protein, acidic and rich in cysteine) is an extracellular, Ca2(+)-binding protein associated with cellular populations undergoing migration, proliferation, and/or differentiation. Active preparations of SPARC bind to specific components of the extracellular matrix and cause mesenchymal cells to assume a rounded phenotype. In this study we show that SPARC modulates the progression of bovine aortic endothelial cells through the cell cycle. At a concentration of 20 micrograms/ml, SPARC inhibited the incorporation of [3H]thymidine into newly synthesized DNA by approximately 70%, as compared to control cultures within 24 hr after the release from G0 phase. The effect was dose-dependent and reached greater than 90% inhibition at 30 micrograms of SPARC per ml after 24 hr. A 20-residue synthetic peptide (termed 2.1) from a non-Ca2(+)-binding, disulfide-rich domain of SPARC also exhibited a dose-dependent inhibition of [3H]thymidine uptake in endothelial cells within 24 hr after release from G0 phase. An inhibition of 50% was seen with peptide 2.1 at a 0.4 mM concentration. Peptides from other regions of the SPARC protein did not produce this effect. Maximum inhibition of [3H]thymidine uptake by SPARC and peptide 2.1 occurred during the early-to-middle G1 phase of the endothelial-cell cycle. From 0-12 hr after release from G0 phase, cells exhibited delayed entry into S phase, which normally occurred at 24 +/- 2 hr. These results were further corroborated by flow cytometry. In the presence of SPARC at 20 micrograms/ml, 72% fewer cells were in S phase after a 24-hr period; a similar, but less marked, reduction was seen with peptide 2.1. Peptide 2.1 did not cause cell rounding, whereas peptide 1.1, a highly efficient inhibitor of endothelial-cell spreading, exhibited essentially no activity with respect to cell-cycle progression. It therefore appears that the transient, inhibitory effect of SPARC on the entry of endothelial cells into S phase does not depend on the overt changes in cell shape mediated through cytoskeletal rearrangement.


Institutional corruption of pharmaceuticals and the myth of safe and effective drugs.
Over the past 35 years, patients have suffered from a largely hidden epidemic of side effects from drugs that usually have few offsetting benefits. The pharmaceutical industry has corrupted the practice of medicine through its influence over what drugs are developed, how they are tested, and how medical knowledge is created. Since 1906, heavy commercial influence has compromised congressional legislation to protect the public from unsafe drugs. The authorization of user fees in 1992 has turned drug companies into the FDA's prime clients, deepening the regulatory and cultural capture of the agency. Industry has demanded shorter average review times and, with less time to thoroughly review evidence, increased hospitalizations and deaths have resulted. Meeting the needs of the drug companies has taken priority over meeting the needs of patients. Unless this corruption of regulatory intent is reversed, the situation will continue to deteriorate. We offer practical suggestions including: separating the funding of clinical trials from their conduct, analysis, and publication; independent FDA leadership; full public funding for all FDA activities; measures to discourage R&D on drugs with few, if any, new clinical benefits; and the creation of a National Drug Safety Board.


A cascade of degradative hydrolase activity contributes to hepatocyte necrosis during anoxia.
Calpain proteases contribute to hepatocyte necrosis during anoxia. Our aim was to ascertain the mechanism causing calpain activation during anoxia. In rat hepatocytes, a twofold increase in calpain activity occurred despite the lack of an increase in cytosolic Ca2+ concentration ([Ca2+]i). The increase in calpain activity was not associated with an increase in calpain mRNA or a decrease in calpastatin mRNA expression. Because phospholipid degradation products generated by phospholipases can activate calpains at physiological [Ca2+]i, we determined the effect of phospholipase inhibitors and activators on calpain activity. Pretreatment of hepatocytes with fluphenazine, a phospholipase inhibitor, decreased calpain activation and improved cell survival. Melittin, a phospholipase A2 activator, increased calpain activity and potentiated cell killing. Finally, phospholipid degradation preceded the increase in calpain activity. Thus the enhanced calpain activity occurring in hepatocytes during anoxia 1) is regulated at the posttranslational level and 2) appears to be dependent on phospholipase activity. These data suggest a novel cascade for degradative hydrolase activity during hepatocyte necrosis by anoxia with phospholipase-mediated activation of calpains.


The genomic and clinical landscape of fetal akinesia.
Fetal akinesia has multiple clinical subtypes with over 160 gene associations, but the genetic etiology is not yet completely understood. In this study, 51 patients from 47 unrelated families were analyzed using next-generation sequencing (NGS) techniques aiming to decipher the genomic landscape of fetal akinesia (FA). We have identified likely pathogenic gene variants in 37 cases and report 41 novel variants. Additionally, we report putative pathogenic variants in eight cases including nine novel variants. Our work identified 14 novel disease-gene associations for fetal akinesia: ADSSL1, ASAH1, ASPM, ATP2B3, EARS2, FBLN1, PRG4, PRICKLE1, ROR2, SETBP1, SCN5A, SCN8A, and ZEB2. Furthermore, a sibling pair harbored a homozygous copy-number variant in TNNT1, an ultrarare congenital myopathy gene that has been linked to arthrogryposis via Gene Ontology analysis. Our analysis indicates that genetic defects leading to primary skeletal muscle diseases might have been underdiagnosed, especially pathogenic variants in RYR1. We discuss three novel putative fetal akinesia genes: GCN1, IQSEC3 and RYR3. Of those, IQSEC3, and RYR3 had been proposed as neuromuscular disease-associated genes recently, and our findings endorse them as FA candidate genes. By combining NGS with deep clinical phenotyping, we achieved a 73% success rate of solved cases.


Effects of changing inspiratory to expiratory time ratio on carbon dioxide elimination during high-frequency jet ventilation.
Clinical observations indicate that gas exchange during high-frequency ventilation can be improved by decreasing the inspiratory to expiratory time ratio (I/E). Decreasing I/E at a fixed frequency may therefore allow a reduction of tidal volume (VT) without affecting gas exchange. This decreased VT may result in lower airway pressures, and thereby reduce the incidence of barotrauma. In order to quantify the effects of I/E on gas exchange, the relationship between carbon dioxide elimination and VT was studied in 6 mongrel dogs at I/E of 1/2, 1/1, and 2/1. Ventilation was provided by a jet-type high-frequency ventilator using a frequency of 5 Hz and VT of 17 to 100% of the anatomic dead space. Carbon dioxide elimination was found to vary inversely with (I/E)0.9, corroborating the clinical observations. These data add further evidence that I/E may be a useful parameter in optimizing high-frequency ventilation and provide a quantitative relationship that can be used to predict the effect that altering I/E will have on gas exchange.


A novel mutant allele of the CYP2A6 gene (CYP2A6*11 ) found in a cancer patient who showed poor metabolic phenotype towards tegafur.
In a clinical study, a newly developed anticancer drug, TS-1 capsule, which contained tegafur (FT) and 5-chloro-2,4-dihydroxypyridine, an inhibitor of dihydropyrimidine dehydrogenase, was orally administered to five gastric cancer patients (patients 1-5). The total area under the plasma FT concentration-time curve in patient 1 was four-fold higher than in other patients. Since cytochrome P450 2A6 (CYP2A6) has been reported to metabolize FT to yield 5-fluorouracil (5-FU), it was postulated that the poor metabolic phenotype of patient 1 was caused by mutations of the CYP2A6 gene. Thus, alleles for the CYP2A6 genes derived from patient 1 were completely sequenced. It was found that one allele was CYP2A6*4C, which was a whole deleted allele for the human CYP2A6 gene. The other allele was a novel mutant allele (CYP2A6*11) in which thymine at nucleotide 670 was changed to cytosine. The nucleotide change caused an amino acid change from serine at residue 224 to proline. To examine whether or not the amino acid change affected CYP2A6 activity, we expressed an intact or mutant CYP2A6 together with NADPH-P450 oxidoreductase in Escherichia coli, and compared the capacity of the wild and mutant enzymes to metabolize FT to 5-FU. The Vmax value for FT metabolism by the mutant CYP2A6 was approximately one-half of the value of the intact CYP2A6, although the Km values were nearly the same. From these results, we conclude that the poor metabolic phenotype of patient 1 was caused by the existence of the two mutant alleles, CYP2A6*4C and the new variant CYP2A6*11.


Central gastrin inhibits feeding behavior and food passage in neonatal chicks.
The gastrin/cholecystokinin (CCK) family is recognized as the principal family of hormones involved in regulation of the gastrointestinal tract CCK is recognized as a satiety hormone in mammalian species, but it has been suggested that gastrin rather CCK may have an important role in controlling feeding behavior in the neonatal chick through a poorly developed blood brain barrier. So far, however, there is no direct evidence that central gastrin inhibits food intake in neonatal chicks. The aim of this study was to elucidate whether central administration of gastrin 1) inhibits feeding behavior and 2) alters food passage from the crop. The effects of central administration of gastrin on food intake were investigated in experiment 1. Birds (2-day-old) were food-deprived for 3 h and then gastrin or saline was injected intracerebroventricularly. Gastrin strongly inhibited food intake in a dose-dependent fashion for 2 h. Thereafter, the effects of central gastrin on feeding behavior and serum corticosterone concentration were examined in experiment 2. Following central administration of gastrin, food intake was depressed and pecking behavior was inhibited. Serum corticosterone concentration was not altered by central administration of gastrin. The influence of central gastrin on food passage from the crop was investigated in experiment 3. Central administration of gastrin clearly delayed food passage. In conclusion, central gastrin appears to have a strong effect for the satiety and gastrointestinal motility in the neonatal chick.


A genome-wide association study of resistance to stripe rust (Puccinia striiformis f. sp. tritici) in a worldwide collection of hexaploid spring wheat (Triticum aestivum L.).
New races of Puccinia striiformis f. sp. tritici (Pst), the causal pathogen of wheat stripe rust, show high virulence to previously deployed resistance genes and are responsible for large yield losses worldwide. To identify new sources of resistance we performed a genome-wide association study (GWAS) using a worldwide collection of 1000 spring wheat accessions. Adult plants were evaluated under field conditions in six environments in the western United States, and seedlings were tested with four Pst races. A single-nucleotide polymorphism (SNP) Infinium 9K-assay provided 4585 SNPs suitable for GWAS. High correlations among environments and high heritabilities were observed for stripe rust infection type and severity. Greater levels of Pst resistance were observed in a subpopulation from Southern Asia than in other groups. GWAS identified 97 loci that were significant for at least three environments, including 10 with an experiment-wise adjusted Bonferroni probability < 0.10. These 10 quantitative trait loci (QTL) explained 15% of the phenotypic variation in infection type, a percentage that increased to 45% when all QTL were considered. Three of these 10 QTL were mapped far from previously identified Pst resistance genes and QTL, and likely represent new resistance loci. The other seven QTL mapped close to known resistance genes and allelism tests will be required to test their relationships. In summary, this study provides an integrated view of stripe rust resistance resources in spring wheat and identifies new resistance loci that will be useful to diversify the current set of resistance genes deployed to control this devastating disease.


Outcomes of medical emergencies on commercial airline flights.
Worldwide, 2.75 billion passengers fly on commercial airlines annually. When in-flight medical emergencies occur, access to care is limited. We describe in-flight medical emergencies and the outcomes of these events. We reviewed records of in-flight medical emergency calls from five domestic and international airlines to a physician-directed medical communications center from January 1, 2008, through October 31, 2010. We characterized the most common medical problems and the type of on-board assistance rendered. We determined the incidence of and factors associated with unscheduled aircraft diversion, transport to a hospital, and hospital admission, and we determined the incidence of death. There were 11,920 in-flight medical emergencies resulting in calls to the center (1 medical emergency per 604 flights). The most common problems were syncope or presyncope (37.4% of cases), respiratory symptoms (12.1%), and nausea or vomiting (9.5%). Physician passengers provided medical assistance in 48.1% of in-flight medical emergencies, and aircraft diversion occurred in 7.3%. Of 10,914 patients for whom postflight follow-up data were available, 25.8% were transported to a hospital by emergency-medical-service personnel, 8.6% were admitted, and 0.3% died. The most common triggers for admission were possible stroke (odds ratio, 3.36; 95% confidence interval [CI], 1.88 to 6.03), respiratory symptoms (odds ratio, 2.13; 95% CI, 1.48 to 3.06), and cardiac symptoms (odds ratio, 1.95; 95% CI, 1.37 to 2.77). Most in-flight medical emergencies were related to syncope, respiratory symptoms, or gastrointestinal symptoms, and a physician was frequently the responding medical volunteer. Few in-flight medical emergencies resulted in diversion of aircraft or death; one fourth of passengers who had an in-flight medical emergency underwent additional evaluation in a hospital. (Funded by the National Institutes of Health.).


An interferometric approach to the measurement of optical polarization.
After discussing the desirability of determining the variation of polarization with frequency in planetary spectra, the possibility of measuring the intensity and state of polarization of optical radiation by means of the high resolution Fourier spectroscopic method is discussed. In the proposed experimental arrangement a two-beam interferometer is used with a polarizer in each beam. After recombination the emergent radiation is analyzed with a linear polarizer. It is shown that the interferograms obtained in this way contain information about the four Stokes parameters of the incident radiation. The polarizers introduce an asymmetry in the interferograms requiring full (exponential) transforms for retrieval of the desired data. The effects of the finite range of path difference and the variation of its zero point with frequency are considered, and evaluation of the corresponding phase error with a proper choice of the polarizer settings is discussed. The formalism also takes into account the residual polarization introduced by the beam splitter, and the differential transmission of the two beams. Generally, three independent interferograms are needed for determining the phase error and the four Stokes parameters. Some simple arrangements are described in which the two beams are either both linearly or both circularly polarized. It is hoped that instruments based on the principle described here will be built by workers in the field.


An assay to image neuronal microtubule dynamics in mice.
Microtubule dynamics in neurons play critical roles in physiology, injury and disease and determine microtubule orientation, the cell biological correlate of neurite polarization. Several microtubule binding proteins, including end-binding protein 3 (EB3), specifically bind to the growing plus tip of microtubules. In the past, fluorescently tagged end-binding proteins have revealed microtubule dynamics in vitro and in non-mammalian model organisms. Here, we devise an imaging assay based on transgenic mice expressing yellow fluorescent protein-tagged EB3 to study microtubules in intact mammalian neurites. Our approach allows measurement of microtubule dynamics in vivo and ex vivo in peripheral nervous system and central nervous system neurites under physiological conditions and after exposure to microtubule-modifying drugs. We find an increase in dynamic microtubules after injury and in neurodegenerative disease states, before axons show morphological indications of degeneration or regrowth. Thus increased microtubule dynamics might serve as a general indicator of neurite remodelling in health and disease.


Oxidation of protein disulfide bonds by singlet oxygen gives rise to glutathionylated proteins.
Disulfide bonds play a key function in determining the structure of proteins, and are the most strongly conserved compositional feature across proteomes. They are particularly common in extracellular environments, such as the extracellular matrix and plasma, and in proteins that have structural (e.g. matrix) or binding functions (e.g. receptors). Recent data indicate that disulfides vary markedly with regard to their rate of reaction with two-electron oxidants (e.g. HOCl, ONOOH), with some species being rapidly and readily oxidized. These reactions yielding thiosulfinates that can react further with a thiol to give thiolated products (e.g. glutathionylated proteins with glutathione, GSH). Here we show that these 'oxidant-mediated thiol-disulfide exchange reactions' also occur during photo-oxidation reactions involving singlet oxygen (1O2). Reaction of protein disulfides with 1O2 (generated by multiple sensitizers in the presence of visible light and O2), yields reactive intermediates, probably zwitterionic peroxyl adducts or thiosulfinates. Subsequent exposure to GSH, at concentrations down to 2 μM, yields thiolated adducts which have been characterized by both immunoblotting and mass spectrometry. The yield of GSH adducts is enhanced in D2O buffers, and requires the presence of the disulfide bond. This glutathionylation can be diminished by non-enzymatic (e.g. tris-(2-carboxyethyl)phosphine) and enzymatic (glutaredoxin) reducing systems. Photo-oxidation of human plasma and subsequent incubation with GSH yields similar glutathionylated products with these formed primarily on serum albumin and immunoglobulin chains, demonstrating potential in vivo relevance. These reactions provide a novel pathway to the formation of glutathionylated proteins, which are widely recognized as key signaling molecules, via photo-oxidation reactions.


Survey of characteristics and treatment preferences for physicians treating postpartum depression in the general medical setting.
This research examines the relationship between the characteristics of physicians who routinely provide medical care for postpartum mothers and their treatment preferences for managing postpartum depression (PPD) by means of a self-administered postal survey. A survey was sent to a random sample of 600 obstetricians/gynecologists and 600 family practitioners in North Carolina. The overall response rate was 42%. Forty-six percent of the responding physicians (N = 228) reported that they had seen women for postpartum visits during the past 3 months. Physician age, gender, race, and practice type were significantly associated with differences in their treatment preferences of PPD. Older physicians and non-white physicians were less likely to prefer treatment of PPD with antidepressants. Older physicians and physicians trained in obstetrics/gynecology (OB/GYN) were less likely to treat PPD with referral to a social worker or psychologist for counseling, but physicians trained in family practice and female physicians were more likely to do so.


African American women's exposure to interpersonal racial discrimination in public settings and preterm birth: the effect of coping behaviors.
To determine whether African American women's coping behaviors modify the relationship between exposure to interpersonal racial discrimination in public settings and preterm birth (<37 weeks). A case-control study was performed among African American women delivering infants at two tertiary care hospitals in Chicago, IL between July 2001-June 2005. A structured questionnaire was administered to measure maternal perceived exposure to interpersonal racial discrimination in public settings and coping behaviors. A greater percentage of African American mothers of preterm infants had high lifetime and past year exposure to racism in public settings than their peers who deliver term infants; odds ratios (OR) and 95% confidence intervals (95% CI) equaled 1.5 (0.9-2.8) for lifetime and 2.5 (1.2-5.2) for past year exposure. Active coping, especially "working harder to prove them wrong" led to attenuated ORs (interaction P value<.05 for lifetime and <.10 for past year. African American women's exposure to racism in public settings is a risk factor for preterm birth; active coping behaviors weaken this relationship.


[Current clinical trials using new targeted therapies for myeloid leukemia and the research trends].
Great progress on insight into genetic aberrations of myeloid leukemia via gene expression profiling has led to better understanding of the pathobiology of this heterogeneous disorder. It enabled the development of specific treatment modalities targeted to underlying oncogenic abnormalities, with well established examples of all-trans retinoic acid for the treatment of acute promyelocytic leukemia and imatinib for chronic myeloid leukemia. However, these strategies have not been completely developed yet in that most of brand new targeted therapies have been somewhat far from achieving cure of leukemia and that many problems with regards to drug resistance and recurrence from minimal residual disease remain to be solved. On the other hand, concept of cancer(leukemic) stem cell and its niche has been shedding new light on oncological field these days. This review summarizes the current clinical trials using new targeted therapies and research trends on myeloid leukemia.


Telomere maintenance requires the RAD51D recombination/repair protein.
The five RAD51 paralogs (RAD51B, RAD51C, RAD51D, XRCC2, and XRCC3) are required in mammalian cells for normal levels of genetic recombination and resistance to DNA-damaging agents. We report here that RAD51D is also involved in telomere maintenance. Using immunofluorescence labeling, electron microscopy, and chromatin immunoprecipitation assays, RAD51D was shown to localize to the telomeres of both meiotic and somatic cells. Telomerase-positive Rad51d(-/-) Trp53(-/-) primary mouse embryonic fibroblasts (MEFs) exhibited telomeric DNA repeat shortening compared to Trp53(-/-) or wild-type MEFs. Moreover, elevated levels of chromosomal aberrations were detected, including telomeric end-to-end fusions, a signature of telomere dysfunction. Inhibition of RAD51D synthesis in telomerase-negative immortalized human cells by siRNA also resulted in telomere erosion and chromosome fusion. We conclude that RAD51D plays a dual cellular role in both the repair of DNA double-strand breaks and telomere protection against attrition and fusion.


Accuracy and Usability of a Novel Algorithm for Detection of Irregular Pulse Using a Smartwatch Among Older Adults: Observational Study.
Atrial fibrillation (AF) is often paroxysmal and minimally symptomatic, hindering its diagnosis. Smartwatches may enhance AF care by facilitating long-term, noninvasive monitoring. This study aimed to examine the accuracy and usability of arrhythmia discrimination using a smartwatch. A total of 40 adults presenting to a cardiology clinic wore a smartwatch and Holter monitor and performed scripted movements to simulate activities of daily living (ADLs). Participants' clinical and sociodemographic characteristics were abstracted from medical records. Participants completed a questionnaire assessing different domains of the device's usability. Pulse recordings were analyzed blindly using a real-time realizable algorithm and compared with gold-standard Holter monitoring. The average age of participants was 71 (SD 8) years; most participants had AF risk factors and 23% (9/39) were in AF. About half of the participants owned smartphones, but none owned smartwatches. Participants wore the smartwatch for 42 (SD 14) min while generating motion noise to simulate ADLs. The algorithm determined 53 of the 314 30-second noise-free pulse segments as consistent with AF. Compared with the gold standard, the algorithm demonstrated excellent sensitivity (98.2%), specificity (98.1%), and accuracy (98.1%) for identifying irregular pulse. Two-thirds of participants considered the smartwatch highly usable. Younger age and prior cardioversion were associated with greater overall comfort and comfort with data privacy with using a smartwatch for rhythm monitoring, respectively. A real-time realizable algorithm analyzing smartwatch pulse recordings demonstrated high accuracy for identifying pulse irregularities among older participants. Despite advanced age, lack of smartwatch familiarity, and high burden of comorbidities, participants found the smartwatch to be highly acceptable.


Relationship between vapor intrusion and human exposure to trichloroethylene.
Trichloroethylene (TCE) in groundwater has the potential to volatilize through soil into indoor air where it can be inhaled. The purpose of this study was to determine whether individuals living above TCE-contaminated groundwater are exposed to TCE through vapor intrusion. We examined associations between TCE concentrations in various environmental media and TCE concentrations in residents. For this assessment, indoor air, outdoor air, soil gas, and tap water samples were collected in and around 36 randomly selected homes; blood samples were collected from 63 residents of these homes. Additionally, a completed exposure survey was collected from each participant. Environmental and blood samples were analyzed for TCE. Mixed model multiple linear regression analyses were performed to determine associations between TCE in residents' blood and TCE in indoor air, outdoor air, and soil gas. Blood TCE concentrations were above the limit of quantitation (LOQ; ≥ 0.012 µg L(-1)) in 17.5% of the blood samples. Of the 36 homes, 54.3%, 47.2%, and >84% had detectable concentrations of TCE in indoor air, outdoor air, and soil gas, respectively. Both indoor air and soil gas concentrations were statistically significantly positively associated with participants' blood concentrations (P = 0.0002 and P = 0.04, respectively). Geometric mean blood concentrations of residents from homes with indoor air concentrations of >1.6 µg m(-3) were approximately 50 times higher than geometric mean blood TCE concentrations in participants from homes with no detectable TCE in indoor air (P < .0001; 95% CI 10.4-236.4). This study confirms the occurrence of vapor intrusion and demonstrates the magnitude of exposure from vapor intrusion of TCE in a residential setting.


Examining the safety and efficacy of a chemotherapy dosing method in Allogeneic Stem Cell Transplant patients of extreme body size.
There is no consensus on a universal dosing method for calculating high-dose chemotherapy in allogeneic Stem Cell Transplant (SCT) patients. The Metropolitan Life (Met-Life) Insurance Company's weight-height tables have been used to determine body weight for chemotherapy dosing for SCT, however no formal study has been done to determine if the Met-Life weight- height tables can be used for chemotherapy dosing in SCT. We retrospectively studied the use of Met-Life weight-height tables for chemotherapy dosing in SCT. Our goal is to determine if patients with extremes of body size who had undergone an SCT and were dosed according to the Met-Life weight- height tables had an increase of Treatment Related Morbidity (TRM) or mortality or relapse. Patients were grouped into three different treatment regimens, cyclophosphamide/TBI, busulphan/cyclophosphamide, and AraC/cyclophosphamide/TBI. Patients in each treatment regimen were further divided into five equal groups based on weight. Treatment related morbidity and mortality was evaluated by comparing the lowest and highest quintiles to the middle quintiles within each treatment regimen. Data from 262 patients was evaluated in this study. Overall, there was not an increase in TRM or mortality or in relapse in patients with extremes of body size. The Met-Life weight-height tables could be used to dose patients undergoing allogeneic SCTs. Additional prospective studies would need to be done comparing other chemotherapy dosing methods with the Met-Life weight-height tables to further validate this conclusion.


The factors affecting persistent pneumothorax and mortality in neonatal pneumothorax.
The aim of this study was to present our experience in management of neonatal pneumothorax and factors contributing to persistent pneumothorax and mortality. Forty-two newborns were analyzed according to gestational age, birth weight, Apgar score, age of admittance, type of delivery, mother's age, side of pneumothorax, causes of pneumothorax, accompanying disorders, tube thoracostomy and mechanical ventilation durations, mean hospital stay, and deaths. Sixteen patients (38%) weighed less than 2500 g and 28 (66%) were preterm. The mean Apgar score at 5th minute was 6.2 (2-10). The pneumothorax was bilateral in 9 patients (21%). There was a defined underlying lung pathology in 26 (61%) patients and accompanying disorder in 14 (33%). Mean tube thoracostomy duration was 5 days (2-12). Twenty-five patients (59%) needed mechanical ventilation. Overall 10 babies died. Our findings indicated that underlying primary lung pathology, need for mechanical ventilation, and bilateral pneumothorax were major determinants of persistent pneumothorax and mortality in newborns.


Dementia care in rural and remote settings: a systematic review of formal or paid care.
The purpose of this review is to critically evaluate the available evidence from the published scientific literature on dementia care and service provision in rural and remote settings from the perspective of formal/paid caregiving, in order to assess the current state of knowledge, identify policy and practice implications, and make recommendations for future research. A systematic review of the literature indexed in ISI Web of Knowledge, PsychInfo, Medline, Healthstar, CINAHL, EMBASE, and Sociological Abstracts was conducted. Data were extracted from papers meeting inclusion criteria: peer-reviewed papers that focused on dementia or Alzheimer's disease (AD), examined care or service provision in relation to persons with AD or dementia, and relevant to rural or remote care or services. The search identified 872 articles for review, reduced to 72 after removing duplicates and articles not meeting criteria. Of the 72 remaining, 46 are included in this current review focusing on formal or paid care. A future review will focus on the 26 studies on informal/unpaid care. Six themes that correspond to the current state of knowledge in rural dementia care in the 46 included studies were: diagnostic processes, service provision, service models and programs, staff education and support needs, use of technology, and long-term care. Despite the growing body of evidence over the 20 years covered by this review, much of the research is descriptive and/or based on small sample sizes, and distributed across the care continuum. Hence the body of evidence on which to base policy and program decisions remains limited. More research is needed that would support the development of comprehensive rural dementia care models.


C-reactive protein and risk of first-ever ischemic and hemorrhagic stroke in a general Japanese population: the Hisayama Study.
The role of high-sensitivity C-reactive protein (hsCRP) in the development of stroke is not clearly understood. We investigated the relationship between serum hsCRP levels and stroke occurrence in a general Japanese population. We followed 2692 subjects > or =40 years of age for 12 years. The relative risks and 95% CIs for ischemic and hemorrhagic stroke occurrence were calculated according to the hsCRP quintiles. During the follow-up, 129 first-ever ischemic and 59 hemorrhagic strokes occurred. In men, the age-adjusted incidence of ischemic stroke significantly increased with elevated serum hsCRP levels; the difference between the first and fifth quintiles was statistically significant (1.4 versus 6.6 per 1000 person-years; P=0.02). This association remained significant even after adjustment for other confounding factors, such as age, systolic blood pressure, ECG abnormalities, diabetes, body mass index, total cholesterol, high-density lipoprotein cholesterol, smoking habits, alcohol intake, and regular exercise (adjusted relative risks, 3.11; 95% CI, 1.04 to 9.32; P=0.04). However, such associations were not observed for ischemic stroke in women or in hemorrhagic stroke in either sex. Among male subjects who were both in the fifth hsCRP level and had hypertension, diabetes, obesity, hypercholesterolemia, or a smoking habit, the risk of ischemic stroke was extremely increased, even after adjustment for other risk factors. Our findings suggest that elevated serum hsCRP levels are an independent risk factor for future ischemic stroke in Japanese men and that the coexistence of a high hsCRP level with another risk factor extremely increases the risk of ischemic stroke.


Comparison of a commercial hexadimethrine bromide method and low-ionic-strength solution for antibody detection with special reference to anti-K.
The sensitivities of manual low-ionic hexadimethrine bromide (Polybrene, LIP) and low-ionic Polybrene indirect antiglobulin tests (LIPAT) were compared with those of a manual low-ionic-strength indirect antiglobulin test (LISS) by using a commercial Polybrene kit. One hundred antibodies were coded, titrated, and tested in parallel. LIP did not detect 36 antibodies: 31 anti-K, two anti-E, two anti-Fya, and one anti-Jka. LIPAT did not detect seven anti-K, two anti-E, and two anti-Jka. The combination of LIP and LIPAT did not detect two anti-E that were reactive only in a two-stage enzyme test and seven anti-K that had titers of 2 or lower by LISS. LISS detected all antibodies except for the two enzyme-reactive anti-E. There were no significant differences in the titers of 63 percent of the antibodies studied. For 54 percent of the antibodies in the Kell system, LISS produced significantly higher titers; for 25 percent of antibodies in the Rh system, LIP did so. The poor sensitivity of the Polybrene kit for anti-K makes it unsuitable as a primary method for antibody screening.


Programming self-organizing multicellular structures with synthetic cell-cell signaling.
A common theme in the self-organization of multicellular tissues is the use of cell-cell signaling networks to induce morphological changes. We used the modular synNotch juxtacrine signaling platform to engineer artificial genetic programs in which specific cell-cell contacts induced changes in cadherin cell adhesion. Despite their simplicity, these minimal intercellular programs were sufficient to yield assemblies with hallmarks of natural developmental systems: robust self-organization into multidomain structures, well-choreographed sequential assembly, cell type divergence, symmetry breaking, and the capacity for regeneration upon injury. The ability of these networks to drive complex structure formation illustrates the power of interlinking cell signaling with cell sorting: Signal-induced spatial reorganization alters the local signals received by each cell, resulting in iterative cycles of cell fate branching. These results provide insights into the evolution of multicellularity and demonstrate the potential to engineer customized self-organizing tissues or materials.


Ca2+-dependent exocytosis of L-glutamate by alphaTC6, clonal mouse pancreatic alpha-cells.
Pancreatic islet cells express receptors and transporters for L-glutamate and are thus believed to use L-glutamate as an intercellular signaling molecule. However, the mechanism by which L-glutamate appears in the islets is unknown. In the present study, we investigated whether L-glutamate is secreted through exocytosis by alphaTC6 cells (clonal mouse pancreatic alpha-cells). An appreciable amount of L-glutamate was released from cultured cells after the addition of KCl or A23187 in the presence of Ca2+ and 10 mmol/l glucose in the medium. The KCl-induced glutamate release was significantly reduced when assayed in the absence of Ca2+ or when the cells were pretreated with EGTA-AM. The KCl-induced Ca2+-dependent glutamate release was inhibited approximately 40% by voltage-gated Ca2+ channel blockers, such as nifedipine at 20 micromol/l. The degree of KCl-induced Ca2+-dependent glutamate release was correlated with an increase in intracellular [Ca2+], as monitored by fura-2 fluorescence. Botulinum neurotoxin type E inhibited 55% of the KCl-induced Ca2+-dependent glutamate release, followed by specific cleavage of 25 kDa synaptosomal-associated protein. Furthermore, bafilomycin A1, a specific inhibitor of vacuolar H+-ATPase, inhibited 40% of the KCl-induced Ca2+-dependent glutamate release. Immunoelectronmicroscopy with antibodies against synaptophysin, a marker for neuronal synaptic vesicles and endocrine synaptic-like microvesicles, revealed a large number of synaptophysin-positive clear vesicles in cells. Digitonin-permeabilized cells took up L-glutamate only in the presence of MgATP, which is sensitive to bafilomycin A1 or 3,5-di-tert-butyl-4-hydroxybenzylidene-malononitrile (a proton conductor) but insensitive to either oligomycin or vanadate. From these results, it was concluded that alphaTC6 cells accumulate L-glutamate in the synaptophysin-containing vesicles in an ATP-dependent manner and secrete it through a Ca2+-dependent exocytic mechanism. The Ca2+-dependent glutamate release was also triggered when cells were transferred in the medium containing 1 mmol/l glucose, suggesting that low glucose treatment stimulates the release of glutamate. Our results are consistent with the idea that L-glutamate is secreted by alpha-cells through Ca2+-dependent regulated exocytosis.


A Novel Preservation Solution Containing a JNK Inhibitory Peptide Efficiently Improves Islet Yield for Porcine Islet Isolation.
For islet transplantation, pancreas preservation in University of Wisconsin (UW) solution is associated with disadvantages, such as collagenase inhibition, resulting in poor islet yield and islets with poor viability. In this study, we evaluated a novel preservation solution, the extracellular-type c-Jun N-terminal kinase (JNK) inhibitor-containing (EJ) solution. The EJ solution has high sodium-low potassium composition with low viscosity compared to UW solution. Moreover, EJ solution contains a recently developed JNK inhibitor from our laboratory. We first compared the performance of EJ solution with that of UW solution. Islet yield before and after purification was significantly higher in the EJ group than in the UW group. Second, we compared the performance of EJ solution with that of EJ solution without the JNK inhibitor (EJ-J solution). After pancreas preservation in EJ solution, JNK activity was maintained at a relatively low level during islet isolation. Islet yield before and after purification was significantly higher in the EJ group than in the EJ-J group. After islet transplantation into streptozotocin-induced diabetic mice, blood glucose levels reached the normoglycemic range in 61.5% and 7.7% of diabetic mice in the EJ and EJ-J groups, respectively. Moreover, EJ solution exhibited reduced inhibition of collagenase digestion compared with UW solution. Advantages of EJ solution over UW solution were inhibition of JNK activity and reduced collagenase inhibition. EJ solution may therefore be more suitable for islet isolation than UW solution.


Oligonucleotide-induced alternative splicing of serotonin 2C receptor reduces food intake.
The serotonin 2C receptor regulates food uptake, and its activity is regulated by alternative pre-mRNA splicing. Alternative exon skipping is predicted to generate a truncated receptor protein isoform, whose existence was confirmed with a new antiserum. The truncated receptor sequesters the full-length receptor in intracellular membranes. We developed an oligonucleotide that promotes exon inclusion, which increases the ratio of the full-length to truncated receptor protein. Decreasing the amount of truncated receptor results in the accumulation of full-length, constitutively active receptor at the cell surface. After injection into the third ventricle of mice, the oligonucleotide accumulates in the arcuate nucleus, where it changes alternative splicing of the serotonin 2C receptor and increases pro-opiomelanocortin expression. Oligonucleotide injection reduced food intake in both wild-type and ob/ob mice. Unexpectedly, the oligonucleotide crossed the blood-brain barrier and its systemic delivery reduced food intake in wild-type mice. The physiological effect of the oligonucleotide suggests that a truncated splice variant regulates the activity of the serotonin 2C receptor, indicating that therapies aimed to change pre-mRNA processing could be useful to treat hyperphagia, characteristic for disorders like Prader-Willi syndrome.


The RhoGAP protein DLC-1 functions as a metastasis suppressor in breast cancer cells.
The identification of molecular signatures characteristic of tumor cells that are capable of metastatic spread is required for the development of therapeutic interventions to abrogate this lethal process. To facilitate this, we have previously characterized an experimental system in which the role of candidate metastasis-related genes can be screened and tested. Monoclonal cell lines M4A4 and NM2C5 are spontaneously occurring sublines of the MDA-MB-435 cell breast tumor cell line that exhibit phenotypic differences in growth, invasion, and metastatic efficiency in athymic mice. In this study, transcriptional profiles of these cell lines were created using oligonucleotide microarrays representing over 12,000 genes. Intensity modeling and hierarchical clustering analysis identified a 171-gene expression signature that correlated with metastatic phenotype and highlighted several GTPase signaling components. Restoration of one of these GTPases, deleted in liver cancer-1 (DLC-1), in metastatic M4A4 cells to levels observed in the nonmetastatic NM2C5 cell line resulted in the inhibition of migration and invasion in vitro and a significant reduction in the ability of these cells to form pulmonary metastases in athymic mice. These studies show the utility of expression profiling, in an appropriate experimental system, to identify genetic determinants of metastatic sufficiency. The finding that DLC-1 can act as a metastasis-suppressor gene supports an influential role for GTPase signaling in tumor progression.


Predictors of high-intensity running capacity in collegiate women during a soccer game.
The purpose of this investigation was to determine which physiological assessments best predicted high-intensity running (HIR) performance during a women's collegiate soccer game. A secondary purpose was to examine the relationships among physiological performance measures including muscle architecture on soccer performance (distance covered, HIR, and sprints during the game) during a competitive collegiate women's soccer game. Ten National Collegiate Athletic Association (NCAA) Division I women soccer players performed physiological assessments within a 2-week period before a competitive regulation soccer game performed during the spring season. Testing consisted of height, body mass, ultrasound measurement of dominant (DOMleg), and nondominant leg (NDOMleg) vastus lateralis for muscle thickness (MT) and pennation angle (PA), VO2max, running economy, and Wingate anaerobic test (WAnT) for peak power (PP), mean power (MP), and fatigue rate (FR). During the game, distance run, HIR, and sprints were measured using a 10-Hz global positioning system. Stepwise regression revealed that VO2max, dominant leg thickness, and dominant leg PA were the strongest predictors of HIR distance during the game (R = 0.989, SEE = 115.5 m, p = 0.001). V[Combining Dot Above]O2max was significantly correlated with total distance run (r = 0.831; p = 0.003), HIR (r = 0.755; p = 0.012), WAnTPP (r = -0.737; p = 0.015), WAnTPP·kg (r = -0.706; p = 0.022), and WAnTFR (r = -0.713; p = 0.021). DOMlegMT was significantly correlated with WAnTFR (r = 0.893; p = 0.001). DOMlegPA was significantly correlated with WAnTFR (r = 0.740; p = 0.023). The NDOMlegPA was significantly correlated to peak running velocity (r = 0.781; p = 0.013) and WAnT MP·kg (r = 0.801; p = 0.01). Results of this study indicate that V[Combining Dot Above]O2max and muscle architecture are important characteristics of NCAA Division I women soccer players and may predict HIR distance during a competitive contest.


Designing spaces for the networked learning landscape.
The concept of the learning landscape is used to explore the range of learning environments needed at multiple scales to better align with changes in the medical education curriculum. Four key scales that correspond to important types of learning spaces are identified: the classroom, the building, the campus and the city. "In-between" spaces are identified as growing in importance given changing patterns of learning and the use of information technology. Technology is altering how learning takes place in a wider variety of types of spaces as it is interwoven into every aspect of learning. An approach to planning learning environments which recognizes the need to think of networks of learning spaces connected across multiple scales is proposed. The focus is shifted from singular spaces to networks of inter-connected virtual and digital environments. A schematic model comprising the networked learning landscape, intended as a guide to planning that emphasizes relationships between the changing curriculum and its alignment with learning environments at multiple scales is proposed in this work. The need for higher levels of engagement of faculty, administrators and students in defining the briefs for the design of new kinds of medical education environments is highlighted.


Clinal variation for only some phenological traits across a species range.
Phenology is the timing of life cycle events of an organism. Alterations in phenology can have profound effects on individual fitness, population growth, and community dynamics. Recent changes in climate have altered the phenology of many organisms, which may result in selection to shift phenological traits. Understanding the relationship between local climates and population differentiation in phenology will allow us to anticipate responses to novel selective environments caused by global climate change. We evaluated population differentiation in the number of days to germination, first flower, and fruit maturation for 33 populations throughout the range of Campanulastrum americanum (American Bellflower). Germination and fruit maturation had geographical clines with earlier timing in populations from northern latitudes. Northern sites were cooler and drier, suggesting potential adaptive differentiation of the shorter life cycle associated with earlier phenology. Similarly, higher elevations were cooler and had earlier fruit maturation. However, seed germination was later in higher elevation populations. Although there was substantial variation in the day to first flower, ranging 40 days between population means, it was idiosyncratic and not related to latitude, suggesting differentiation in response to selective factors distinct from those on germination and fruit maturation. Thus, germination and fruit maturation in C. americanum may shift in response to selection by rising temperatures. However, such changes are not expected for flowering time, a typical indicator of climate change.


Safe drinking water in regional NSW, Australia.
The New South Wales (NSW) Public Health Act 2010 requires water suppliers to implement a drinking water quality assurance program that addresses the 'Framework for management of drinking water quality' in the Australian drinking water guidelines. NSW Health has recognised the importance of a staged implementation of this requirement and the need to support regional water utilities. To date, NSW Health has assisted 74 regional utilities to develop and implement their management systems. The Public Health Act 2010 has increased awareness of drinking water risk management, and offers a systematic process to identify and control risks. This has benefited large utilities, smaller suppliers, and remote and Aboriginal communities. Work is continuing to ensure implementation of the process by private suppliers and water carters.


Conserved UBE3A subcellular distribution between human and mice is facilitated by non-homologous isoforms.
The human UBE3A gene, which is essential for normal neurodevelopment, encodes three Ubiquitin E3 ligase A (UBE3A) protein isoforms. However, the subcellular localization and relative abundance of these human UBE3A isoforms are unknown. We found, as previously reported in mice, that UBE3A is predominantly nuclear in human neurons. However, this conserved subcellular distribution is achieved by strikingly distinct cis-acting mechanisms. A single amino-acid deletion in the N-terminus of human hUBE3A-Iso3, which is homologous to cytosolic mouse mUBE3A-Iso2, results in its translocation to the nucleus. This singe amino-acid deletion is shared with apes and Old World monkeys and was preceded by the appearance of the cytosolic hUBE3A-Iso2 isoform. This hUBE3A-Iso2 isoform arose after the lineage of New World monkeys and Old World monkeys separated from the Tarsiers (Tarsiidae). Due to the loss of a single nucleotide in a non-coding exon, this exon became in frame with the remainder of the UBE3A protein. RNA-seq analysis of human brain samples showed that the human UBE3A isoforms arise by alternative splicing. Consistent with the predominant nuclear enrichment of UBE3A in human neurons, the two nuclear-localized isoforms, hUBE3A-Iso1 and -Iso3, are the most abundantly expressed isoforms of UBE3A, while hUBE3A-Iso2 maintains a small pool of cytosolic UBE3A. Our findings provide new insight into UBE3A localization and evolution and may have important implications for gene therapy approaches in Angelman syndrome.


"Death by a Thousand Cuts": Agriculture Producer Resiliency in the Western United States.
Agricultural producers face a wide array of stress triggers, shocks, and long-term pressures such as drought, flooding, fire, government policies, financial insecurity, and physical injuries. Extant research has revealed that mental health stigma, lack of access to care in rural areas, and negative coping responses (alcohol abuse, suicide, prescription drugs use) exacerbate the challenge of producer responses to short and long-term adversity. Resilience, the traits, processes, and capacities of producers to adapt and transform their approach to farming or ranching, when necessary, in response to stress triggers or long-term pressures, has received less research attention, particularly in the Western United States. The purpose of the study was to apply an interactionist occupational resilience theoretical perspective to the investigation of contextual factors contributing to resilience in Western United States agricultural producers. Qualitative interviews (45 to 90 minutes) were conducted with agricultural producers (n=51) from Western states and territories. Applied thematic analysis with a phenomenological lens was utilized to analyze interview transcriptions. First and second level coding were conducted to derive themes. The analysis revealed that resilience is based upon the interactions between traits of producers and the context of agriculture. Four themes were generated (Agricultural Life, External Stressors, Traits and Adaptations, and Supports and Resources), supported by subthemes. The themes and subthemes are depicted in an agricultural producer resiliency model. The findings shed light on the equivocal role of neighbors in providing support for each other and the double-edged sword of co-working with family. The findings underscore that social capital is an important mechanism for supporting farmers and ranchers, as those with stronger social resources are more resilient. We recommend more funding to tailor stress and mental health programming to the specifics of agriculture, integration of behavioral health in primary care as a mechanism to increase access to care, and more intentional technical assistance for farmers and ranchers on strategic planning and problem solving.


GPR56/ADGRG1 is associated with response to antidepressant treatment.
It remains unclear why many patients with depression do not respond to antidepressant treatment. In three cohorts of individuals with depression and treated with serotonin-norepinephrine reuptake inhibitor (N = 424) we show that responders, but not non-responders, display an increase of GPR56 mRNA in the blood. In a small group of subjects we also show that GPR56 is downregulated in the PFC of individuals with depression that died by suicide. In mice, we show that chronic stress-induced Gpr56 downregulation in the blood and prefrontal cortex (PFC), which is accompanied by depression-like behavior, and can be reversed by antidepressant treatment. Gpr56 knockdown in mouse PFC is associated with depressive-like behaviors, executive dysfunction and poor response to antidepressant treatment. GPR56 peptide agonists have antidepressant-like effects and upregulated AKT/GSK3/EIF4 pathways. Our findings uncover a potential role of GPR56 in antidepressant response.


Expansion of cytolytic CD8(+) natural killer T cells with limited capacity for graft-versus-host disease induction due to interferon gamma production.
T cells with natural killer cell phenotype and function (NKT cells) have been described in both human and murine tissues. In this study, culture conditions were developed that resulted in the expansion of CD8(+) NKT cells from bone marrow, thymus, and spleen by the timed addition of interferon-gamma (IFN-gamma), interleukin 2 (IL-2), and anti-CD3 monoclonal antibody. After 14 to 21 days in culture, dramatic expansion of CD3(+), CD8(+), alphabetaT-cell receptor(+) T cells resulted with approximately 20% to 50% of the cells also expressing the NK markers NK1.1 and DX5. The CD8(+) NKT cells demonstrated lytic activity against several tumor target cells with more than 90% lysis by day 14 to day 21 of culture. Cytotoxicity was observed against both syngeneic and allogeneic tumor cell targets with the greatest lytic activity by the cells expressing either NK1.1 or DX5. The expanded CD8(+) NKT cells produce T(H)1-type cytokines with high levels of IFN-gamma and tumor necrosis factor alpha. Expansion of the CD8(+) NKT cells was independent of CD1d. Ly49 molecules were expressed on only a minority of cells. A single injection of expanded CD8(+) NKT cells was capable of protecting syngeneic animals from an otherwise lethal dose of Bcl1 leukemia cells. Expanded CD8(+) NKT cells produced far less graft-versus-host disease (GVHD) than splenocytes across major histocompatibility barriers, even when 10 times the number of CD8(+) NKT cells as compared to splenocytes were injected. This reduction in GVHD was related to IFN-gamma production since cells expanded from IFN-gamma knock-out animals caused acute lethal GVHD, whereas cells expanded from animals defective in fas ligand, fas, IL-2, and perforin did not. These data indicate that CD8(+) NKT cells expanded in this fashion could be useful for preserving graft-versus-leukemia activity without causing GVHD.


Problem based medical education in the community: a student nutritional survey in Nigeria.
Students in their second year of medical studies perceived nutritional deficiency as a possible health problem in rural Nigeria and conducted a survey to investigate and compare the nutritional and health status of three distinct groups of children (Bokos in Babana, Bokos in Marami, Fulani). The nutritional status of the children in the three study populations differed and seasonal change affected the one to four year old children among the Fulani more than the other 2 groups. An attempt was made to explain the results on the basis of environmental and socioeconomic differences among the groups. This account, more than just elaborating on figures and more than making indisputable statements, aims at showing how medical students, from their earliest studies, can learn from an apparent health problem of the community, how they can be messengers of data relevant to picture the local epidemiology (practical aspect) and how their findings can be used as self instructive teaching--learning material (educational aspect).


Porcine sialoadhesin: a newly identified xenogeneic innate immune receptor.
Extracorporeal porcine liver perfusion is being developed as a bridge to liver allotransplantation for patients with fulminant hepatic failure. This strategy is limited by porcine Kupffer cell destruction of human erythrocytes, mediated by lectin binding of a sialic acid motif in the absence of antibody and complement. Sialoadhesin, a macrophage restricted lectin that binds sialic acid, was originally described as a sheep erythrocyte binding receptor. Given similarities between sialoadhesin and the unidentified macrophage lectin in our model, we hypothesized porcine sialoadhesin contributed to recognition of human erythrocytes. Two additional types of macrophages were identified to bind human erythrocytes-spleen and alveolar. Expression of sialoadhesin was confirmed by immunofluorescence in porcine tissues and by flow cytometry on primary macrophages. A stable transgenic cell line expressing porcine sialoadhesin (pSn CHO) bound human erythrocytes, while a sialoadhesin mutant cell line did not. Porcine macrophage and pSn CHO recognition of human erythrocytes was inhibited approximately 90% by an antiporcine sialoadhesin monoclonal antibody and by human erythrocyte glycoproteins. Furthermore, this binding was substantially reduced by sialidase treatment of erythrocytes. These data support the hypothesis that porcine sialoadhesin is a xenogeneic receptor that mediates porcine macrophage binding of human erythrocytes in a sialic acid-dependent manner.