Can Chinese Medicine Be Used for Prevention of Corona Virus Disease 2019 (COVID-19)? A Review of Historical Classics, Research Evidence and Current Prevention Programs.
Since December 2019, an outbreak of corona virus disease 2019 (COVID-19) occurred in Wuhan, and rapidly spread to almost all parts of China. This was followed by prevention programs recommending Chinese medicine (CM) for the prevention. In order to provide evidence for CM recommendations, we reviewed ancient classics and human studies. Historical records on prevention and treatment of infections in CM classics, clinical evidence of CM on the prevention of severe acute respiratory syndrome (SARS) and H1N1 influenza, and CM prevention programs issued by health authorities in China since the COVID-19 outbreak were retrieved from different databases and websites till 12 February, 2020. Research evidence included data from clinical trials, cohort or other population studies using CM for preventing contagious respiratory virus diseases. The use of CM to prevent epidemics of infectious diseases was traced back to ancient Chinese practice cited in Huangdi's Internal Classic (Huang Di Nei Jing) where preventive effects were recorded. There were 3 studies using CM for prevention of SARS and 4 studies for H1N1 influenza. None of the participants who took CM contracted SARS in the 3 studies. The infection rate of H1N1 influenza in the CM group was significantly lower than the non-CM group (relative risk 0.36, 95% confidence interval 0.24-0.52; n=4). For prevention of COVID-19, 23 provinces in China issued CM programs. The main principles of CM use were to tonify qi to protect from external pathogens, disperse wind and discharge heat, and resolve dampness. The most frequently used herbs included Radix astragali (Huangqi), Radix glycyrrhizae (Gancao), Radix saposhnikoviae (Fangfeng), Rhizoma Atractylodis Macrocephalae (Baizhu), Lonicerae Japonicae Flos (Jinyinhua), and Fructus forsythia (Lianqiao). Based on historical records and human evidence of SARS and H1N1 influenza prevention, Chinese herbal formula could be an alternative approach for prevention of COVID-19 in high-risk population. Prospective, rigorous population studies are warranted to confirm the potential preventive effect of CM.


Relaxation and guided imagery in Hispanic persons diagnosed with fibromyalgia: a pilot study.
Fibromyalgia (FM) is a chronic pain disorder affecting 2% of the population in the United States. Another 16 million Hispanics suffer from generically identified rheumatic diseases that likely include FM. Because there are few reported studies of Hispanics with FM, a pilot study using a repeated-measures pretest-posttest design investigated the effects of a 10-week mind-body intervention (visual imagery with relaxation) on symptom management. The change in means from baseline to week 10 demonstrated improvement in self-efficacy for managing pain and other symptoms and functional status. Visual imagery with relaxation is a mind-body intervention that may be used for symptom management in this population.


Validating the interval to biochemical failure for the identification of potentially lethal prostate cancer.
To validate the interval to biochemical failure (IBF) as a prognostic factor at the time of biochemical failure for prostate cancer mortality (PCM) following radiotherapy (RT). From a collaborative data set of men with clinically localized prostate cancer treated with RT from four institutions in three countries, we identified 1,722 men with biochemical failure (BF; prostate-specific antigen nadir + 2 ng/mL). The IBF was defined as the time interval from completion of treatment to the date of BF. The primary outcome measure was discriminatory power in the form of the concordance index (c-index). Seventeen percent of men had an IBF ≤ 18 months. Median potential follow-up beyond the time of BF was 67 months. There were 290 deaths from prostate cancer. The IBF was the most discriminating individual prognostic factor overall, with a sensitivity of IBF ≤ 18 months to predict PCM within 10 years of 48.4% (95% CI, 43.3% to 54.1%); the specificity was 86.1% (95% CI, 84.5% to 87.7%), equating to a c-index of 0.611 (95% CI, 0.578 to 0.647). The 5-year cumulative incidence of PCM for IBF more than 18 months versus IBF ≤ 18 months was 9.4% (95% CI, 7.7% to 11.5%) versus 26.3% (95% CI, 21.2% to 31.8%); corresponding 10-year estimates were 26.2% (95% CI, 21.5% to 30.8%) versus 55.9% (95% CI, 48.9% to 63.0%), respectively (P < .001 for both). IBF exhibited minimal change in performance across various follow-up durations. IBF is the single most robust prognostic factor for PCM following RT without androgen deprivation therapy. This external validation demonstrates that patients and clinicians can use this information to make decisions about subsequent treatments.


Towards a human brain EV atlas: Characteristics of EVs from different brain regions, including small RNA and protein profiles.
Extracellular vesicles (EVs) are released from different cell types in the central nervous system (CNS) and play roles in regulating physiological and pathological functions. Although brain-derived EVs (bdEVs) have been successfully collected from brain tissue, there is not yet a "bdEV atlas" of EVs from different brain regions. To address this gap, we separated EVs from eight anatomical brain regions of a single individual and subsequently characterized them by count, size, morphology, and protein and RNA content. The greatest particle yield was from cerebellum, while the fewest particles were recovered from the orbitofrontal, postcentral gyrus, and thalamus regions. EV surface phenotyping indicated that CD81 and CD9 were more abundant than CD63 for all regions. Cell-enriched surface markers varied between brain regions. For example, putative neuronal markers NCAM, CD271, and NRCAM were more abundant in medulla, cerebellum, and occipital regions, respectively. These findings, while restricted to tissues from a single individual, suggest that additional studies are merited to lend more insight into the links between EV heterogeneity and function in the CNS.


Effective Management of Peritoneal Dialysis-Associated Hydrothorax in a Child: A Case Report.
Peritoneal dialysis (PD) confers many advantages, including a better quality of life for children with end-stage renal disease; however, the procedure is associated with several complications, including pleuroperitoneal leaks. Here, we report an unusual case of hydrothorax caused by long-term PD in a child, which was further complicated by pneumonia. A 9-year-old boy who had received CAPD for 22 months presented with dyspnea, swelling, and increased body weight. Chest tube drainage yielded 500 mL of transudative fluid. Computed tomography peritoneography revealed increased outflow from the peritoneum to the pleural cavity. PD was suspended, and hemodialysis (HD) was initiated. Video-assisted thoracoscopic surgery was performed; however, because the patient had pneumonia during hospitalization, pleural adhesions with a septated appearance occurred. This resulted in difficulties identifying pleuroperitoneal fistula (PPF). Right pleural effusion resolved following pleurodesis using bleomycin. Regular HD was performed for 10 weeks, and PD was subsequently reinitiated. There was no recurrence of hydrothorax during long-term follow-up. We suspect that the underlying mechanism of hydrothorax in our patient was associated with a PPF that formed either due to a congenital diaphragmatic defect or an acquired defect, resulting in dialysate leakage. Our case demonstrates that a temporary switch from PD to HD, accompanied by pleurodesis, may help resolve hydrothorax that occurs as a complication of long-term PD.


[Postpartum hemorrhage--an update].
Postpartum hemorrhage remains a major cause of maternal morbidity and mortality. The incidence of postpartum hemorrhage appears to be increasing in developed countries due to an increased number of placenta accreta or percreta after previous Cesarean deliveries. The initial therapy of postpartum hemorrhage consists of uterotonic drugs and inspection of the uterine cavum. At the same time, optimization of the clotting potential should be initiated early. Tranexamic acid may be considered as a first line choice, followed by fibrinogen if necessary. If bleeding continues, fresh frozen plasma and packed red cells should be ordered in a ratio of 1:1, as this ratio has been shown to improve survival in trauma victims. All labor and delivery suites should have standard operating procedures for the management of postpartum hemorrhage in place with regular drills.


Long-chain acyl-CoA profiles in cultured fibroblasts from patients with defects in fatty acid oxidation.
Negative chemical ionization (NCI) mass spectrometry was used to quantify the acyl-CoA intermediates present in human fibroblasts growing in media containing the long-chain fatty acid, palmitate. The acyl-CoA intermediates were detected as the N-acyl pentafluorobenzyl glycinates. In fibroblasts from normal individuals only saturated acyl-CoA esters were detected, supporting the concept that the acyl-CoA dehydrogenase reaction is the rate-limiting step of intramitochondrial fatty acid oxidation. In patients with inherited enzymatic defects of intramitochondrial long-chain fatty acid oxidation, there was not a significant increase in the amount of long-chain acyl-CoA compounds, with palmitoyl-CoA amounts similar to those found in controls. However, there was a sharp decrease in the relative amount of lauroyl-CoA and a resultant sixfold elevation in the palmitoyl-CoA:lauroyl-CoA ratio. In contrast, fibroblasts with a defect involving the transport of fatty acids across the mitochondrial membrane, carnitine palmitoyl transferase 1 deficiency, had a fourfold increase in palmitoyl-CoA. Our results suggest that acyl-CoA esters in biological tissues are readily detectable using NCI mass spectrometry. This approach is significantly more sensitive than previous methods for the detection of these important metabolic intermediates, and may prove useful in the study of fatty acid oxidation in both normal and enzyme-deficient tissues.


Anti-inflammatory effects of retinoids and carotenoid derivatives on caspase-3-dependent apoptosis and efferocytosis of bovine neutrophils.
To evaluate immunomodulatory properties of all-trans retinoic acid and a fully oxidized β-carotene dietary product in calves with Mannheimia haemolytica-induced pneumonia. Twenty-five 6- to 10-week-old male Holstein calves for experimental inoculations and three 8- to 30-week-old Angus heifers for blood donations. In vitro, neutrophils and monocyte-derived macrophages isolated from blood of healthy Angus heifers were treated with all-trans retinoic acid (1 μM) or fully oxidized β-carotene (8.3 μg/mL) for various times and assessed for markers of cellular death, antimicrobial function, and production of proinflammatory leukotriene B4. Following 28 days of dietary supplementation with fully oxidized β-carotene, Holstein calves were experimentally inoculated with M haemolytica. Bronchoalveolar lavage fluid was collected at 3 and 24 hours after challenge inoculation and analyzed for markers of apoptosis. In vitro, all-trans retinoic acid and fully oxidized β-carotene induced cell-selective, caspase-3-dependent apoptosis in neutrophils, which subsequently enhanced efferocytosis in macrophages. Conversely, neither treatment altered phorbol 12-myristate 13-acetate-induced oxidative burst, phagocytosis of nonopsonized zymosan (complement or antibody independent), or M haemolytica-induced leukotriene B4 production in bovine neutrophils. In vivo, fully oxidized β-carotene enhanced leukocyte apoptosis in bronchoalveolar lavage fluid as well as subsequent efferocytosis by macrophages without altering numbers of circulating leukocytes. Neutrophil apoptosis and subsequent efferocytosis by macrophages are key mechanisms in the resolution of inflammation. Findings for the present study indicated that all-trans retinoic acid and fully oxidized β-carotene could be novel nutraceutical strategies that may confer anti-inflammatory benefits for cattle with respiratory tract disease.


ElectroMotive drug administration (EMDA) of Mitomycin C as first-line salvage therapy in high risk "BCG failure" non muscle invasive bladder cancer: 3 years follow-up outcomes.
In case of high grade non-muscle invasive bladder cancer (HG-NMIBC), intravesical BCG represents the first-line treatment; despite the "gold" standard therapy, up to 50% of patients relapse, needing radical cystectomy. Hence, alternative therapeutic strategies have been developed. The aim of the study was to evaluate a first-line salvage treatment with EMDA®-MMC in patients with HGNMIBC unresponsive to BCG. We carried out a prospective, single-center, single-arm Phase II study in order to evaluate the efficacy (in terms of recurrence and progression) and the safety of the EMDA®-MMC treatment in 26 (21 male, 5 female) consecutive patients with "BCG refractory" HGNMIBC on a 3 years follow-up. EMDA®-MMC treatment consisted of 40 mg of MMC diluted in 100 ml of sterile water retained in the bladder for 30 min with 20 mA pulsed electric current. EMDA®-MMC regimen consisted of an induction course of 6 weekly instillations followed by a maintenance course of 6 monthly instillations. Follow-up was performed with systematic mapping biopsies of the bladder (with sampling in the prostatic urethra for men), voiding and washing urinary cytology, radiological study of the upper urinary tract. We performed Survival Kaplan-Meier curves and Log-rank test in order to analyze high grade disease-free survival. At the end of follow-up, 16 patients (61.5%) preserved their native bladder; 10 patients (38.4%) underwent radical cystectomy, in 6 patients (23.1%) for recurrent HGNMIBC and in 4 patients (15.4%) for progression to muscle-invasive disease. At the end of follow-up, stratifying patients based on TNM classification (TaG3, T1G3, Cis, TaT1G3 + Cis), disease-free rates were 75, 71.4, 50 and 25%, respectively; survival curves showed statistically significant differences (p value < 0.05). Regarding toxicity, we reported severe adverse systemic event of hypersensitivity to the MMC in 3 patients (11.5%), and local side effects in 6 patients (26.1%). In the field of alternative strategies to radical cystectomy, the EMDA®-MMC could be considered safe and effective in high-risk NMIBC unresponsive to BCG, as a "bladder sparing" therapy in selected patients. Multicenter studies with a larger number of patients and a longer follow-up might confirm our preliminary results. EudraCT2017-002585-43. 17 June 2017 (retrospectively registered).


HLA class II transgenic mice as models of human diseases.
Predisposition to develop various autoimmune disorders has been associated with certain HLA class II molecules but there is a lack of information on the pathophysiological role of HLA genes in conferring susceptibility. Various experimental animal models of autoimmune disease have been studied to address the role of immune response genes. To study the interactions involved between class II molecules (DQ and DR) and define the immunologic mechanisms in various diseases, we generated HLA-DR and DQ transgenic mice that lacked endogenous class II molecules. The HLA molecules in these mice are expressed on the cell surface and can positively select CD4+ T cells expressing various V beta T-cell receptors (TCR). A peripheral tolerance is maintained to transgenic HLA molecules thus indicating that these molecules act as self. Mouse co-stimulatory and accessory molecules can interact with the HLA-peptide-TCR complex leading to efficient T-cell activation. In this review, we describe immunogenetic models for human diseases using these transgenic mice. Our studies show that HLA class II transgene-restricted T cells recognize the immunodominant antigens and peptide epitopes, similar to HLA class II-restricted human T cells. Thus these mice provide powerful tools to understand the role of HLA class II molecules in predisposition and onset of human diseases and to develop immunotherapy and vaccines.


Extreme context specificity in differential allelic expression.
Variability in cis-regulation of gene expression has been implicated in the phenotypic manifestation of complex traits including common, multifactorial diseases. The differential expression of alleles due to polymorphism in cis-regulatory elements is common in the human genome, but there is a paucity of information about the context specificity of these control elements. In this study, we examined the differential allelic expression (DAE) of BMP5 in human mesenchymal tissues obtained from 16 donors undergoing joint replacement for treatment of osteoarthritis. We observed significant differences in BMP5 allelic output, with allelic ratios greater than 4:1 (P < 10(-20)) in the tissues of some donors. We also discovered a significant variability in allelic expression within the different tissues of donors. For 12 of our donors, we examined the allelic expression of BMP5 in two different regions of cartilage: cartilage adjacent to the site of the osteoarthritic lesion and cartilage distal from the lesion. Five of these 12 donors demonstrated highly significant differences (P < or = 10(-8)) in allelic expression between the different regions of their cartilage. Using DAE as a phenotype, we attempted to map tissue-specific cis-regulatory polymorphisms, and we identified a single nucleotide polymorphism located downstream of BMP5, which was significantly associated with DAE in some but not all of the examined tissues. These findings suggest that allelic expression can be highly context specific and that when interrogating the cis-regulatory control of a particular gene, one cannot necessarily assume that allelic expression is conserved across different tissues or even across different regions of the same tissue.


Insulin-dependent diabetes mellitus and the major histocompatibility complex peptide transporters TAP1 and TAP2: no association in a population with a high disease incidence.
Although many studies have established an association between insulin-dependent diabetes mellitus (IDDM) and the class II region of the human major histocompatibility complex (MHC), it has been difficult to assign susceptibility to a single locus. Recently, two antigen-processing genes, TAP1 and TAP2, have been identified within the region. Previous studies have reached conflicting conclusions as to the role of these genes in IDDM; it is uncertain whether an increased frequency of the allele TAP2A and a concomitant decrease in TAP2B are independent disease associations or secondary to linkage disequilibrium (LD) between TAP2A and HLA-DR3. To further investigate this question, we have characterized TAP1 and TAP2 alleles in 129 IDDM patients from Sardinia, a population with limited genetic heterogeneity and a high disease incidence. When compared to 90 random controls, the only significant difference was a decrease in the minor allele TAP2C in patients. However, when HLA-DR and -DQ matched controls were compared, this difference disappeared. Further analysis suggested that TAP2C was in LD with HLA-DRB1*1401 and subtypes of HLA-DRB1*11, alleles which were not observed in the IDDM population. LD was also observed between other TAP and HLA-DR alleles, in particular between TAP2A and HLA-DR3 in both patients and controls. Our data supports the conclusion that there is no primary association between TAP2 alleles and IDDM, and that previously reported associations may be due to LD with other class II loci.


Prevalence and cost of nonadherence with antiepileptic drugs in an adult managed care population.
This study assessed the extent of refill nonadherence with antiepileptic drugs (AEDs) and the potential association between AED nonadherence and health care costs in an adult-managed care population. Retrospective claims from the PharMetrics database were analyzed. Inclusion criteria were: age > or =21, epilepsy diagnosis between January 01, 2000 and March 12, 2005, > or =2 AED prescriptions, and continuous health plan enrollment for > or =6 months prior to and > or =12 months following AED initiation. Adherence was evaluated using the medication possession ratio (MPR). Patients with an MPR <0.8 were classified as nonadherent. Multivariate regression was used to assess the effect of AED nonadherence on annualized cost outcomes. Regression covariates included patient demographics, Charlson Comorbidity Index (CCI), and follow-up duration. Among patients meeting all inclusion criteria (N = 10,892), 58% were female, mean age was 44 years, mean CCI was 0.94, and mean follow-up was 27 months. Mean MPR was 0.78 and 39% of patients were nonadherent. AED nonadherence was associated with an increased likelihood of hospitalization (odds ratio [OR]= 1.110, p = 0.013) and emergency room (ER) admission (OR = 1.479, p < 0.0001), as well as increased inpatient and ER costs of $1,799 and $260 (both p = 0.001), respectively, per patient per year. Outpatient and other ancillary costs were not significantly affected by nonadherence. A large net positive effect of nonadherence on total annual health care costs remained (+$1,466, p = 0.034) despite an offset from reduced prescription drug intake. Adherence with AEDs among adult epilepsy patients is suboptimal and nonadherence appears to be associated with increased health care costs. Efforts to promote AED adherence may lead to cost savings for managed care systems.


Salt intake, asthma, and exercise-induced bronchoconstriction: a review.
Despite the heterogeneous treatment options for patients with asthma, there remains a substantial burden of unaddressed disease, even with optimal treatment. Epidemiological studies indicate that patients frequently resort to complimentary and alternative therapies when being treated for asthma and other chronic health conditions. Changes in diet associated with the development of a more affluent lifestyle is one of the environmental factors considered to contribute to the increased prevalence of asthma in the past few decades. Dietary sodium in particular has been considered to be a dietary constituent implicated in this phenomenon. This article reviews the studies conducted that have questioned whether reducing dietary salt intake potentially improves pulmonary function and airway hyper-responsiveness in asthmatics, as well as studies evaluating dietary salt intake on the severity of exercise-induced bronchoconstriction (EIB). The data presented supporting dietary salt restriction for reducing airway hyper-responsiveness in asthmatics is encouraging, though not clinically convincing. Studies conducted previously have been limited for a variety of reasons, including limitations related to the experiment and populations studied. However, in studies that evaluated the severity of EIB in asthmatic individuals and involved altered dietary salt intake, data have been more convincing. A low-sodium diet maintained for 1 to 2 weeks decreases bronchoconstriction in response to exercise in individuals with asthma. There are no data regarding the longer-term effects of a low-sodium diet on either the prevalence or severity of asthma or on EIB. As a low-sodium diet has other beneficial health effects, it can be considered a therapeutic option for adults with asthma, although it should be considered as an adjunctive intervention to supplement optimal pharmacotherapy, and not as an alternative.


Characteristics of American Indian Female Caregivers on a Southwest American Indian Reservation.
American Indian (AI) caregivers have been excluded from national survey efforts. Drawing from a 2012 survey administered on the Hopi Reservation in northern Arizona, 20% of adults are caregivers. More information is needed to guide program development tailored to Hopi needs. In a University-Community collaboration, a 58 question survey was administered to self-identified caregivers of a family member about amount and type of care provided, difficulties, caregiver health, and desired support services. Characteristics of caregivers and their experiences were described. Forty-four (44) female Hopi caregivers were interviewed from June-October 2017, mean age of 59 years (± 12.6) with mean 5.5 year (± 4.4) history of providing care. Over 84% provided care to either a parent or grandparent. Most caregivers provided transportation (93.2%), housework (93.2%), and medical related care (72.7%). Caregivers stated they had difficulties with not having enough time for family and or friends (88.6%), financial burdens (75.0%), and not having enough time for themselves (61.4%). The most frequently identified difficulty was stress (45.5%). Caregivers would like additional services, with 76.7% asking for training. Over 77% would not consider placing their relative in an assisted living facility. Compared to national data, Hopi female caregivers are older, provide more care hours/week, more caregiving duties, and for a longer number of years. Stress is the most reported difficulty, although lower than national levels. As caregivers are resistant to placing the recipient in assisted living, educational efforts should focus on training caregivers to assist the care recipient and decreasing caregiver stress.


Comparisons of intensity-duration patterns of physical activity in the US, Jamaica and 3 African countries.
This difference in how populations living in low-, middle or upper-income countries accumulate daily PA, i.e. patterns and intensity, is an important part in addressing the global PA movement. We sought to characterize objective PA in 2,500 participants spanning the epidemiologic transition. The Modeling the Epidemiologic Transition Study (METS) is a longitudinal study, in 5 countries. METS seeks to define the association between physical activity (PA), obesity and CVD risk in populations of African origin: Ghana (GH), South Africa (SA), Seychelles (SEY), Jamaica (JA) and the US (suburban Chicago). Baseline measurements of objective PA, SES, anthropometrics and body composition, were completed on 2,500 men and women, aged 25-45 years. Moderate and vigorous PA (MVPA, min/d) on week and weekend days was explored ecologically, by adiposity status and manual labor. Among the men, obesity prevalence reflected the level of economic transition and was lowest in GH (1.7%) and SA (4.8%) and highest in the US (41%). SA (55%) and US (65%) women had the highest levels of obesity, compared to only 16% in GH. More men and women in developing countries engaged in manual labor and this was reflected by an almost doubling of measured MPVA among the men in GH (45 min/d) and SA (47 min/d) compared to only 28 min/d in the US. Women in GH (25 min/d), SA (21 min/d), JA (20 min/d) and SEY (20 min/d) accumulated significantly more MPVA than women in the US (14 min/d), yet this difference was not reflected by differences in BMI between SA, JA, SEY and US. Moderate PA constituted the bulk of the PA, with no study populations except SA men accumulating > 5 min/d of vigorous PA. Among the women, no sites accumulated >2 min/d of vigorous PA. Overweight/obese men were 22% less likely to engage in manual occupations. While there is some association for PA with obesity, this relationship is inconsistent across the epidemiologic transition and suggests that PA policy recommendations should be tailored for each environment.


Cell biology of the leaf epidermis: Fate specification, morphogenesis, and coordination.
As the outermost layer of plants, the epidermis serves as a critical interface between plants and the environment. During leaf development, the differentiation of specialized epidermal cell types, including stomatal guard cells, pavement cells, and trichomes, occurs simultaneously, each providing unique and pivotal functions for plant growth and survival. Decades of molecular-genetic and physiological studies have unraveled key players and hormone signaling specifying epidermal differentiation. However, most studies focus on only one cell type at a time, and how these distinct cell types coordinate as a unit is far from well-comprehended. Here we provide a review on the current knowledge of regulatory mechanisms underpinning the fate specification, differentiation, morphogenesis, and positioning of these specialized cell types. Emphasis is given to their shared developmental origins, fate flexibility, as well as cell cycle and hormonal controls. Furthermore, we discuss computational modeling approaches to integrate how mechanical properties of individual epidermal cell types and entire tissue/organ properties mutually influence each other. We hope to illuminate the underlying mechanisms coordinating the cell differentiation that ultimately generate a functional leaf epidermis.


Versatile virus-like particle carrier for epitope based vaccines.
Recombinant proteins and in particular single domains or peptides are often poorly immunogenic unless conjugated to a carrier protein. Virus-like-particles are a very efficient means to confer high immunogenicity to antigens. We report here the development of virus-like-particles (VLPs) derived from the RNA bacteriophage AP205 for epitope-based vaccines. Peptides of angiotensin II, S.typhi outer membrane protein (D2), CXCR4 receptor, HIV1 Nef, gonadotropin releasing hormone (GnRH), Influenza A M2-protein were fused to either N- or C-terminus of AP205 coat protein. The A205-peptide fusions assembled into VLPs, and peptides displayed on the VLP were highly immunogenic in mice. GnRH fused to the C-terminus of AP205 induced a strong antibody response that inhibited GnRH function in vivo. Exposure of the M2-protein peptide at the N-terminus of AP205 resulted in a strong M2-specific antibody response upon immunization, protecting 100% of mice from a lethal influenza infection. AP205 VLPs are therefore a very efficient and new vaccine system, suitable for complex and long epitopes, of up to at least 55 amino acid residues in length. AP205 VLPs confer a high immunogenicity to displayed epitopes, as shown by inhibition of endogenous GnRH and protective immunity against influenza infection.


Integrated metagenomic analysis of the rumen microbiome of cattle reveals key biological mechanisms associated with methane traits.
Methane is one of the major contributors to global warming. The rumen microbiota is directly involved in methane production in cattle. The link between variation in rumen microbial communities and host genetics has important applications and implications in bioscience. Having the potential to reveal the full extent of microbial gene diversity and complex microbial interactions, integrated metagenomics and network analysis holds great promise in this endeavour. This study investigates the rumen microbial community in cattle through the integration of metagenomic and network-based approaches. Based on the relative abundance of 1570 microbial genes identified in a metagenomics analysis, the co-abundance network was constructed and functional modules of microbial genes were identified. One of the main contributions is to develop a random matrix theory-based approach to automatically determining the correlation threshold used to construct the co-abundance network. The resulting network, consisting of 549 microbial genes and 3349 connections, exhibits a clear modular structure with certain trait-specific genes highly over-represented in modules. More specifically, all the 20 genes previously identified to be associated with methane emissions are found in a module (hypergeometric test, p<10-11). One third of genes are involved in methane metabolism pathways. The further examination of abundance profiles across 8 samples of genes highlights that the revealed pattern of metagenomics abundance has a strong association with methane emissions. Furthermore, the module is significantly enriched with microbial genes encoding enzymes that are directly involved in methanogenesis (hypergeometric test, p<10-9).


Medicaid Incentives for Preventing Chronic Disease: Effects of Financial Incentives for Smoking Cessation.
To test the effectiveness of financial incentives for smoking cessation in the Medicaid population. Secondary data from the Medicaid Incentives for Prevention of Chronic Disease (MIPCD) program and Medicaid claims/encounter data from 2010 to 2015 for five states. Beneficiaries were randomized into receipt or no receipt of financial incentives. We ran multivariate regression models testing the impact of financial incentives on the use of counseling services, smoking behavior, and Medicaid expenditures and utilization. Participating states provided Medicaid eligibility, claims and encounters, program enrollment, and incentivized service use data. Participants who received incentives were more likely to call the Quitline and complete counseling sessions. Incentive receipt was positively associated with self-reported quit attempts, self-reported quits, or passing cotinine tests of smoking cessation in most programs, although results were only statistically significant in a subset. There was no systematic evidence that incentives affected health care use or spending. Financial incentives are a promising policy lever to motivate behavioral change in the Medicaid population, but more evidence is needed regarding optimal incentive size, effectiveness of process-versus outcome-based incentives, targeting of incentives, and long-run cost-effectiveness.


Accuracy of dentists in the clinical diagnosis of oral lesions.
Dentists play an important role in the early diagnosis and treatment of oral lesions. However, treatment based solely on a clinical impression of the diagnosis, without histologic confirmation, can result in serious consequences, particularly when the lesion is precancerous or cancerous. The purpose of this study was to determine the overall accuracy of clinical diagnoses made by dentists as well as to compare the diagnostic ability of general practitioners with members of various dental specialties. The biopsy reports of 976 specimens submitted to the Department of Oral and Maxillofacial Pathology, Virginia Commonwealth University School of Dentistry, between January 2009 and January 2010 were reviewed. The presumptive clinical diagnosis made by the practitioner and the final histologic diagnosis on each specimen were recorded in addition to whether the submitting dentist was a general practitioner or a specialist. Of the clinical diagnoses made by the submitting dentists, 43% were incorrect. General dentists misdiagnosed 45.9%, oral and maxillofacial surgeons 42.8%, endodontists 42.2%, and periodontists 41.2% of the time. The most commonly missed clinical diagnoses were hyperkeratosis (16%), focal inflammatory fibrous hyperplasia (10%), fibroma (8%), periapical granuloma (7%), and radicular cyst (6%). Cancerous lesions were misdiagnosed 5.6% of the time. The high rates of clinical misdiagnosis by dental practitioners indicate that all excised lesions should to be submitted for histologic diagnosis.


IMRT delivery verification using a spiral phantom.
In this paper we report on the testing and verification of a system for IMRT delivery quality assurance that uses a cylindrical solid water phantom with a spiral trajectory for radiographic film placement. This spiral film technique provides more complete dosimetric verification of the entire IMRT treatment than perpendicular film methods, since it samples a three-dimensional dose subspace rather than using measurements at only one or two depths. As an example, the complete analysis of the predicted and measured spiral films is described for an intracranial IMRT treatment case. The results of this analysis are compared to those of a single field perpendicular film technique that is typically used for IMRT QA. The comparison demonstrates that both methods result in a dosimetric error within a clinical tolerance of 5%, however the spiral phantom QA technique provides a more complete dosimetric verification while being less time consuming. To independently verify the dosimetry obtained with the spiral film, the same IMRT treatment was delivered to a similar phantom in which LiF thermoluminescent dosimeters were arranged along the spiral trajectory. The maximum difference between the predicted and measured TLD data for the 1.8 Gy fraction was 0.06 Gy for a TLD located in a high dose gradient region. This further validates the ability of the spiral phantom QA process to accurately verify delivery of an IMRT plan.


Impacted third molar transplantation on the malpracticed extraction socket.
Autotransplantation with or without endodontic therapy is regarded as an alternative treatment option for the replacement of missing teeth. A primary responsibility of a maxillofacial surgeon is to reverse any malpractice to promote successful outcomes and improve the patient's quality of life. This paper presents a malpractice case of incorrect extraction of the lower second molar instead of the impacted third molar. A simple technique of transplanting the impacted third molar to the site of the extracted second molar is introduced by a maxillofacial specialist in Ghana. By making an intentional root socket and fixation without using additional appliances, a novel second molar was achieved with complete recovery. This patient was followed after transplant for a four-year period with the best satisfaction. A grant of the Korean Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea. (HI15C0689).


Long distance active hyperspectral sensing using high-power near-infrared supercontinuum light source.
A hyperspectral remote sensing instrument employing a novel near-infrared supercontinuum light source has been developed for active illumination and identification of targets. The supercontinuum is generated in a standard normal dispersion multi-mode fiber and has 16 W total optical output power covering 1000 nm to 2300 nm spectral range. A commercial 256-channel infrared spectrometer was used for broadband infrared detection. The feasibility of the presented hyperspectral measurement approach was investigated both indoors and in the field. Reflection spectra from several diffusive targets were successfully measured and a measurement range of 1.5 km was demonstrated.


Variations in bitter-taste receptor genes, dietary intake, and colorectal adenoma risk.
Genetic variants in bitter-taste receptor genes have been hypothesized to negatively impact health outcomes and/or influence dietary intake and, consequently, could increase the risk of colorectal neoplasia. Using a case-control study of 914 colorectal adenoma cases/1188 controls, we explored associations among colorectal adenoma risk, dietary intake, and genetic variation in 3 bitter-taste receptor genes: TAS2R38 (rs713598, rs1726866, rs10246939), TAS2R16 (rs846672), and TAS2R50 (rs1376251). Analysis of covariance was conducted to detect trends in dietary intake across TAS2R genotypes/haplotypes. Odds ratios and 95% confidence intervals were estimated by logistic regression to test gene-adenoma risk associations. No significant associations were observed between the TAS2R38 PAV/PAV diplotype or the TAS2R16 (rs846672) polymorphism with the selected diet variables. We observed weak inverse associations between the TAS2R50 (rs1376251) C allele and dietary fiber and vegetable intake (Ps < 0.015). Odds ratios for adenoma risk were not significantly different from the null. Our findings do not support a link between these TAS2R genotypes/haplotypes and dietary intake that could impact colorectal adenoma risk. However, given the paucity of data, we cannot dismiss the possibility that these genes may influence colorectal adenoma risk in other ways, such as through impaired gastrointestinal function, particularly in subgroups of the population.


Tuning graphene transistors through ad hoc electrostatics induced by a nanometer-thick molecular underlayer.
We report on the modulation of the electrical properties of graphene-based transistors that mirror the properties of a few nanometers thick layer made of dipolar molecules sandwiched in between the 2D material and the SiO2 dielectric substrate. The chemical composition of the films of quinonemonoimine zwitterion molecules adsorbed onto SiO2 has been explored by means of X-ray photoemission and mass spectroscopy. Graphene-based devices are then fabricated by transferring the 2D material onto the molecular film, followed by the deposition of top source-drain electrodes. The degree of supramolecular order in disordered films of dipolar molecules was found to be partially improved as a result of the electric field at low temperatures, as revealed by the emergence of hysteresis in the transfer curves of the transistors. The use of molecules from the same family, which are suitably designed to interact with the dielectric surface, results in the disappearance of the hysteresis. DFT calculations confirm that the dressing of the molecules by an external electric field exhibits multiple minimal energy landscapes that explain the thermally stabilized capacitive coupling observed. This study demonstrates that the design and exploitation of ad hoc molecules as an interlayer between a dielectric substrate and graphene represents a powerful tool for tuning the electrical properties of the 2D material. Conversely, graphene can be used as an indicator of the stability of molecular layers, by providing insight into the energetics of ordering of dipolar molecules under the effect of electrical gating.


Evidence for catch-up in cognition and receptive vocabulary among adolescents born very preterm.
Very preterm adolescents display persistent deficits in neuropsychological functions. To compare cognitive and language outcomes at 16 years and cognitive and receptive vocabulary trajectories throughout school years between very preterm and term children and to determine child and family factors associated with better developmental trajectories. At 8, 12, and 16 years, 322 very preterm children with birth weights of 1250 g or less and 41 term children had cognitive and language testing. Hierarchical growth-curve modeling was used to delineate the differences in cognitive and receptive vocabulary development between participants. Cluster analyses allowed for the characterization of very preterm children with different patterns of cognitive and receptive vocabulary development. At 16 years, very preterm adolescents had deficits in general cognition and higher-order language skills (phonological awareness and phonemic decoding) compared with term peers. Although the between-group difference in cognitive scores remained stable from 8 to 16 years, very preterm children demonstrated catch-up gains in receptive vocabulary during the same period. Moreover, subgroups of very preterm children displayed developmental trajectories in cognition similar to term children (55% on the vocabulary and 46% on the block-design subtests). These children had lower rates of neurosensory impairment and mothers with higher education and were from an ethnic nonminority. Significant catch-up in receptive vocabulary is observed by the age of 16 years among very preterm children compared to term peers. The absence of neurosensory impairment and residing in a favorable socioeconomic milieu are associated with the most optimal developmental trajectories.


Long-term Outcomes After Autologous Hematopoietic Stem Cell Transplantation for Multiple Sclerosis.
Autologous hematopoietic stem cell transplantation (AHSCT) may be effective in aggressive forms of multiple sclerosis (MS) that fail to respond to standard therapies. To evaluate the long-term outcomes in patients who underwent AHSCT for the treatment of MS in a large multicenter cohort. Data were obtained in a multicenter, observational, retrospective cohort study. Eligibility criteria were receipt of AHSCT for the treatment of MS between January 1995 and December 2006 and the availability of a prespecified minimum data set comprising the disease subtype at baseline; the Expanded Disability Status Scale (EDSS) score at baseline; information on the administered conditioning regimen and graft manipulation; and at least 1 follow-up visit or report after transplant. The last patient visit was on July 1, 2012. To avoid bias, all eligible patients were included in the analysis regardless of their duration of follow-up. Data analysis was conducted from September 1, 2014 to April 27, 2015. Demographic, disease-related, and treatment-related exposures were considered variables of interest, including age, disease subtype, baseline EDSS score, number of previous disease-modifying treatments, and intensity of the conditioning regimen. The primary outcomes were MS progression-free survival and overall survival. The probabilities of progression-free survival and overall survival were calculated using Kaplan-Meier survival curves and multivariable Cox proportional hazards regression analysis models. Valid data were obtained from 25 centers in 13 countries for 281 evaluable patients, with median follow-up of 6.6 years (range, 0.2-16 years). Seventy-eight percent (218 of 281) of patients had progressive forms of MS. The median EDSS score before mobilization of peripheral blood stem cells was 6.5 (range, 1.5-9). Eight deaths (2.8%; 95% CI, 1.0%-4.9%) were reported within 100 days of transplant and were considered transplant-related mortality. The 5-year probability of progression-free survival as assessed by the EDSS score was 46% (95% CI, 42%-54%), and overall survival was 93% (95% CI, 89%-96%) at 5 years. Factors associated with neurological progression after transplant were older age (hazard ratio [HR], 1.03; 95% CI, 1.00-1.05), progressive vs relapsing form of MS (HR, 2.33; 95% CI, 1.27-4.28), and more than 2 previous disease-modifying therapies (HR, 1.65; 95% CI, 1.10-2.47). Higher baseline EDSS score was associated with worse overall survival (HR, 2.03; 95% CI, 1.40-2.95). In this observational study of patients with MS treated with AHSCT, almost half of them remained free from neurological progression for 5 years after transplant. Younger age, relapsing form of MS, fewer prior immunotherapies, and lower baseline EDSS score were factors associated with better outcomes. The results support the rationale for further randomized clinical trials of AHSCT for the treatment of MS.


Antipsychotic drugs in pregnancy: a review of their maternal and fetal effects.
Understanding the risks of antipsychotic medication use in pregnancy is becoming an important clinical concern given the evidence of their increasing rate of prescription in the general population for a range of disorders. Despite antipsychotics being amongst the earliest of psychotropic medications to be introduced, the evidence for their effects secondary to pregnancy exposure is extremely limited. While this review does not identify clear evidence for a risk of malformation, there is evidence for risks associated with pregnancy and neonatal outcomes. Studies identified found risks that included prematurity, low and high birth weight, and gestational diabetes. There have also been studies that suggest neonatal withdrawal and abnormal muscles movements. The longer term neurodevelopmental outcomes for children exposed in utero remain unclear with only four studies identified: two of first generation antipsychotics and two of second generation antipsychotics. When considering the risk of these medications in pregnancy, the risk of untreated maternal illness (particularly schizophrenia and bipolar disorder) on both maternal and child outcomes is relevant. Future research needs to focus on prospective, longitudinal studies with adequate measures of key confounding variables including maternal mental illness, other exposures (such as smoking, alcohol and illicit drug use) and adequate length of follow up where accurate child developmental measures are obtained.


RecA and RadA proteins of Brucella abortus do not perform overlapping protective DNA repair functions following oxidative burst.
Very little is known about the role of DNA repair networks in Brucella abortus and its role in pathogenesis. We investigated the roles of RecA protein, DNA repair, and SOS regulation in B. abortus. While recA mutants in most bacterial species are hypersensitive to UV damage, surprisingly a B. abortus recA null mutant conferred only modest sensitivity. We considered the presence of a second RecA protein to account for this modest UV sensitivity. Analyses of the Brucella spp. genomes and our molecular studies documented the presence of only one recA gene, suggesting a RecA-independent repair process. Searches of the available Brucella genomes revealed some homology between RecA and RadA, a protein implicated in E. coli DNA repair. We considered the possibility that B. abortus RadA might be compensating for the loss of RecA by promoting similar repair activities. We present functional analyses that demonstrated that B. abortus RadA complements a radA defect in E. coli but could not act in place of the B. abortus RecA. We show that RecA but not RadA was required for survival in macrophages. We also discovered that recA was expressed at high constitutive levels, due to constitutive LexA cleavage by RecA, with little induction following DNA damage. Higher basal levels of RecA and its SOS-regulated gene products might protect against DNA damage experienced following the oxidative burst within macrophages.


[(18)F]Fluorobenzoyllysinepentanedioic Acid Carbamates: New Scaffolds for Positron Emission Tomography (PET) Imaging of Prostate-Specific Membrane Antigen (PSMA).
Radiolabeled urea-based low-molecular weight inhibitors of the prostate-specific membrane antigen (PSMA) are under intense investigation as imaging and therapeutic agents for prostate and other cancers. In an effort to provide agents with less nontarget organ uptake than the ureas, we synthesized four (18)F-labeled inhibitors of PSMA based on carbamate scaffolds. 4-Bromo-2-[(18)F]fluorobenzoyllysineoxypentanedioic acid (OPA) carbamate [(18)F]23 and 4-iodo-2-[(18)F]fluorobenzoyllysine OPA carbamate [(18)F]24 in particular exhibited high target-selective uptake in PSMA+ PC3 PIP tumor xenografts, with tumor-to-kidney ratios of >1 by 4 h postinjection, an important benchmark. Because of its high tumor uptake (90% injected dose per gram of tissue at 2 h postinjection) and high tumor-to-organ ratios, [(18)F]23 is promising for clinical translation. Prolonged tumor-specific uptake demonstrated by [(18)F]24, which did not reach equilibrium during the 4 h study period, suggests carbamates as alternative scaffolds for mitigating dose to nontarget tissues.


Atypical residency of short-beaked common dolphins (Delphinus delphis) to a shallow, urbanized embayment in south-eastern Australia.
Short-beaked common dolphins (Delphinus delphis) are typically considered highly mobile, offshore delphinids. This study assessed the residency of a small community of short-beaked common dolphins in the shallow, urbanized Port Phillip Bay, south-eastern Australia. The ability to identify common dolphins by their dorsal fin markings and coloration using photo-identification was also investigated. Systematic and non-systematic boat surveys were undertaken between 2007 and 2014. Results showed that 13 adult common dolphins and their offspring inhabit Port Phillip Bay, of which 10 adults exhibit residency to the bay. The majority of these adults are reproductively active females, suggesting that female philopatry may occur in the community. Systematic surveys conducted between 2012 and 2014 revealed that the dolphins were found in a median water depth of 16 m and median distance of 2.2 km from the coast. The shallow, urbanized habitat of this resident common dolphin community is atypical for this species. As a result, these common dolphins face threats usually associated with inshore bottlenose dolphin communities. We suggest that the Port Phillip Bay common dolphin community is considered and managed separate to those outside the embayment and offshore to ensure the community's long-term viability and residency in the bay.


Noise-enhanced coding in phasic neuron spike trains.
The stochastic nature of neuronal response has lead to conjectures about the impact of input fluctuations on the neural coding. For the most part, low pass membrane integration and spike threshold dynamics have been the primary features assumed in the transfer from synaptic input to output spiking. Phasic neurons are a common, but understudied, neuron class that are characterized by a subthreshold negative feedback that suppresses spike train responses to low frequency signals. Past work has shown that when a low frequency signal is accompanied by moderate intensity broadband noise, phasic neurons spike trains are well locked to the signal. We extend these results with a simple, reduced model of phasic activity that demonstrates that a non-Markovian spike train structure caused by the negative feedback produces a noise-enhanced coding. Further, this enhancement is sensitive to the timescales, as opposed to the intensity, of a driving signal. Reduced hazard function models show that noise-enhanced phasic codes are both novel and separate from classical stochastic resonance reported in non-phasic neurons. The general features of our theory suggest that noise-enhanced codes in excitable systems with subthreshold negative feedback are a particularly rich framework to study.


Design, synthesis, and evaluation of in vivo potency and selectivity of epoxysuccinyl-based inhibitors of papain-family cysteine proteases.
The papain-family cathepsins are cysteine proteases that are emerging as promising therapeutic targets for a number of human disease conditions ranging from osteoporosis to cancer. Relatively few selective inhibitors for this family exist, and the in vivo selectivity of most existing compounds is unclear. We present here the synthesis of focused libraries of epoxysuccinyl-based inhibitors and their screening in crude tissue extracts. We identified a number of potent inhibitors that display selectivity for endogenous cathepsin targets both in vitro and in vivo. Importantly, the selectivity patterns observed in crude extracts were generally retained in vivo, as assessed by active-site labeling of tissues from treated animals. Overall, this study identifies several important compound classes and highlights the use of activity-based probes to assess pharmacodynamic properties of small-molecule inhibitors in vivo.


Voices of the five doctoral training councils in psychology: seeking common ground on Combined-Integrated doctoral training in psychology.
Doctoral training programs in psychology are accredited by the American Psychological Association in four areas: Clinical, Counseling, School, and Combined-Integrated (C-I) psychology. Each area of doctoral preparation in psychology has its own council, which represents its interests within the field and to external constituents. Despite the fact that Combined-Integrated training programs have existed since the mid-1970s, the Consortium representing this area's interests only formed in the past two years. The purpose of this article was to obtain input from representatives of all five training councils regarding the role of the newly formed council in relation to the other councils and the role of Combined-Integrated training. We begin with a brief description of each of the five councils. We then summarize the representatives' responses to narrative questions regarding advantages and disadvantages associated with meeting to discuss the combined model. Finally, common and divergent themes across the councils in training psychologists are presented, and the future role for Combined-Integrated programs is discussed.


Epstein-Barr virus hepatitis: diagnostic value of in situ hybridization, polymerase chain reaction, and immunohistochemistry on liver biopsy from immunocompetent patients.
Epstein-Barr virus (EBV) hepatitis is an uncommon, almost always self-limited disease in immunocompetent patients. Accurate diagnosis is imperative for appropriate clinical management. The aim of this study was to compare 3 available methods for EBV detection on routinely processed liver biopsies to determine their effectiveness in aiding the diagnosis. In 6 of the 8 cases of EBV hepatitis, EBV was detected by both polymerase chain reaction (PCR) for EBV DNA and in situ hybridization (ISH) for EBV early RNA (EBER). EBV was detected by PCR only in 1 case, and by ISH only in another. EBER-positive cells detected by ISH were typically few and individually distributed in the portal tracts and sinusoids. Immunohistochemical staining for EBV latent membrane proteins was negative in all 8 cases. Five cases of chronic hepatitis C used as negative controls were negative by all 3 detection methods for EBV. These data indicate that PCR and ISH are equally sensitive in detecting EBV in routinely processed liver biopsies. The ready implementation of ISH in pathology laboratories makes it a useful ancillary tool in confirming the diagnosis of EBV hepatitis in equivocal cases. However, EBER-positive cells can be sparse and easily overlooked. Immunohistochemistry for EBV latent membrane proteins apparently has no utility in the diagnosis of EBV hepatitis.


Neurodevelopment of children following prenatal exposure to venlafaxine, selective serotonin reuptake inhibitors, or untreated maternal depression.
Effects on child neurodevelopment of neurotransmitter reuptake inhibitors used as antidepressants during pregnancy have not been adequately studied. The authors compared the effects of prenatal exposure to venlafaxine (serotonin-norepinephrine reuptake inhibitor), selective serotonin reuptake inhibitors (SSRIs), and maternal depression. A cohort derived from a prospectively collected database included four groups of children born to 1) depressed women who took venlafaxine during pregnancy (N=62), 2) depressed women who took SSRIs during pregnancy (N=62), 3) depressed women who were untreated during pregnancy (N=54), and 4) nondepressed, healthy women (N=62). The children's intelligence and behavior outcomes were evaluated with standardized instruments at one time point between the ages of 3 years and 6 years, 11 months. The children exposed to venlafaxine, SSRIs, and maternal depression during pregnancy had similar full-scale IQs (105, 105, and 108, respectively). The IQs of the venlafaxine and SSRI groups were significantly lower than that of the children of nondepressed mothers (112). The three groups exposed to maternal depression had consistently, but nonsignificantly, higher rates of most problematic behaviors than the children of nondepressed mothers. Severity of maternal depression in pregnancy and at testing predicted child behavior. Maternal IQ and child sex predicted child IQ. Antidepressant dose and duration during pregnancy did not predict any cognitive or behavioral outcome. Factors other than antidepressant exposure during pregnancy strongly predict children's intellect and behavior. Depression during pregnancy is a significant risk factor for postpartum depression. Children of depressed mothers may be at risk of future psychopathology.


Xenopus spinal neurons express Kv2 potassium channel transcripts during embryonic development.
Developmentally regulated delayed rectifier potassium currents determine the waveform of the action potential in all Xenopus embryonic primary spinal neurons. To examine this developmental program at the molecular level, we have isolated Xenopus Kv2 potassium channel genes Kv2.1 and Kv2.2. Both genes induce functional heterologous expression of delayed rectifier potassium currents. Transcripts from both Kv2 genes are present in developing embryos; however, only Kv2.2 mRNA is detectable in embryonic spinal neurons. Notably, Kv2.2 transcripts localize to ventral spinal neurons, whereas previously described Kv1.1 transcripts are found in dorsal spinal neurons. Thus, spinal neuron subtypes express distinct potassium channel genes, yet they temporally coordinate functional expression of delayed rectifier potassium currents.


Failure of Ploidy and Proliferative Fraction to Predict Long-Term Outcome After Prostatectomy.
Historically, ploidy and S phase percentage appeared to be promising predictors for prostate cancer recurrence. Lack of uniformity and consistency hampered their development. We evaluated ploidy and S phase for prostate cancer death in a cohort of patients with long-term follow-up. We identified 127 patients that had ploidy and S phase determined at the time of their radical prostatectomy for prostate cancer. With 15 years of follow-up, we determined the risk of biochemical failure and risk of death from prostate cancer. We correlated the S phase and ploidy findings with standard pathology findings. A total of 107 (84%) had diploid and 20 (16%) had non-diploid cancers. The median S phase was 6.6%. There was no correlation of ploidy (P = 0.472) or S phase with preoperative PSA or Gleason score. On univariate analysis, EPE, margin positivity, seminal vesicle involvement, lymph node involvement, high Gleason score and PSA > 10 ng/mL were all predictive of biochemical failure. Ploidy and S phase were not. For prostate cancer death, only Gleason score was predictive. With long-term follow-up in our cohort, Gleason score was predictive of prostate cancer death. Ploidy and S phase were not predictive for biochemical failure or prostate cancer mortality.


Inhibitory effects of cannabidiol on voltage-dependent sodium currents.
Cannabis sativa contains many related compounds known as phytocannabinoids. The main psychoactive and nonpsychoactive compounds are Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD), respectively. Much of the evidence for clinical efficacy of CBD-mediated antiepileptic effects has been from case reports or smaller surveys. The mechanisms for CBD's anticonvulsant effects are unclear and likely involve noncannabinoid receptor pathways. CBD is reported to modulate several ion channels, including sodium channels (Nav). Evaluating the therapeutic mechanisms and safety of CBD demands a richer understanding of its interactions with central nervous system targets. Here, we used voltage-clamp electrophysiology of HEK-293 cells and iPSC neurons to characterize the effects of CBD on Nav channels. Our results show that CBD inhibits hNav1.1-1.7 currents, with an IC50 of 1.9-3.8 μm, suggesting that this inhibition could occur at therapeutically relevant concentrations. A steep Hill slope of ∼3 suggested multiple interactions of CBD with Nav channels. CBD exhibited resting-state blockade, became more potent at depolarized potentials, and also slowed recovery from inactivation, supporting the idea that CBD binding preferentially stabilizes inactivated Nav channel states. We also found that CBD inhibits other voltage-dependent currents from diverse channels, including bacterial homomeric Nav channel (NaChBac) and voltage-gated potassium channel subunit Kv2.1. Lastly, the CBD block of Nav was temperature-dependent, with potency increasing at lower temperatures. We conclude that CBD's mode of action likely involves 1) compound partitioning in lipid membranes, which alters membrane fluidity affecting gating, and 2) undetermined direct interactions with sodium and potassium channels, whose combined effects are loss of channel excitability.


Spread of Aedes japonicus japonicus (Theobald, 1901) in Austria, 2011-2015, and first records of the subspecies for Hungary, 2012, and the principality of Liechtenstein, 2015.
The Asian bush mosquito, Aedes (Hulecoeteomyia) japonicus japonicus (Theobald, 1901) (Diptera: Culicidae), was first identified in Austria in August 2011 in the federal state of Styria at the border to Slovenia. Between 2011 and 2015 the spread of Ae. j. japonicus was monitored in southern, eastern and western Austrian provinces as well as in neighbouring countries by checking natural and man-made container habitats for the aquatic stages. The search concentrated around the most recent occurrence of Ae. j. japonicus and extended up to several kilometres until the subspecies could not be found anymore. Between May and July 2012 the distribution area of Ae. j. japonicus was found to be extended westwards into Carinthia, and eastwards towards the federal state of Burgenland. In August 2012, the subspecies was found in Hungary, representing the first record of an invasive mosquito species in this country. In 2013 its expansion was confirmed at several sites in Austria. Additionally, between April and July 2015, the subspecies was detected in all districts of the westernmost Austrian state Vorarlberg reaching the alpine Montafon valley at the end of October 2015, at all three examined sites in southern Bavaria bordering Vorarlberg, and in the adjacent Principality of Liechtenstein, for which it also represents the first record of an invasive mosquito species. One remarkable finding of the subspecies was located close to the city of Kufstein in the lower Inn valley of the Tyrol in September 2015, which is an isolated occurrence without spatial connection to any known established population. Our findings demonstrate the ongoing spread of Ae. j. japonicus towards all directions within Austria and beyond. Together with the absence of supposed natural barriers, e.g. high mountain chains, at the borders of the current subspecies' distribution area in south-eastern Austria, these findings suggest a further spread to the Austrian capital Vienna and the Hungarian tourist region of Lake Balaton within the upcoming few years. The observed intrusions in western Austria represent most probably extensions of the population established and spreading in eastern Switzerland and southern Germany. The putative role of the subspecies in pathogen transmission together with its rapid spread observed argues for the implementation of comprehensive nation-wide surveillance and response preparedness.


Observation of a New Excited Beauty Strange Baryon Decaying to Ξ_{b}^{-}π^{+}π^{-}.
The Ξ_{b}^{-}π^{+}π^{-} invariant mass spectrum is investigated with an event sample of proton-proton collisions at sqrt[s]=13 TeV, collected by the CMS experiment at the LHC in 2016-2018 and corresponding to an integrated luminosity of 140 fb^{-1}. The ground state Ξ_{b}^{-} is reconstructed via its decays to J/ψΞ^{-} and J/ψΛK^{-}. A narrow resonance, labeled Ξ_{b}(6100)^{-}, is observed at a Ξ_{b}^{-}π^{+}π^{-} invariant mass of 6100.3±0.2(stat)±0.1(syst)±0.6(Ξ_{b}^{-}) MeV, where the last uncertainty reflects the precision of the Ξ_{b}^{-} baryon mass. The upper limit on the Ξ_{b}(6100)^{-} natural width is determined to be 1.9 MeV at 95% confidence level. The low Ξ_{b}(6100)^{-} signal yield observed in data does not allow a measurement of the quantum numbers of the new state. However, following analogies with the established excited Ξ_{c} baryon states, the new Ξ_{b}(6100)^{-} resonance and its decay sequence are consistent with the orbitally excited Ξ_{b}^{-} baryon, with spin and parity quantum numbers J^{P}=3/2^{-}.


Increased turnover of muscle contractile proteins in Duchenne muscular dystrophy as assessed by 3-methylhistidine and creatinine excretion.
1. Myofibrillar protein degradation has been measured in patients with Duchenne muscular dystrophy, normal boys, adult males and Duchenne carriers by the rate of 3-methylhistidine excretion after transfer of subjects to a meat-free diet. 2. Although absolute rates of protein breakdown are lower in Duchenne patients, expression of the data to allow for differences in muscle mass gives fractional degradation rates 2--3 times higher than in age-matched controls. 3. Fractional rates of muscle protein synthesis are increased in the Duchenne patients to almost the same extent as protein breakdown. 4. Rates of muscle protein breakdown in obligate and presumed carriers of the Duchenne gene are not different from controls.


BMP2/BMP4 colorectal cancer susceptibility loci in northern and southern European populations.
Genome-wide association studies have successfully identified 20 colorectal cancer susceptibility loci. Amongst these, four of the signals are defined by tagging single nucleotide polymorphisms (SNPs) on regions 14q22.2 (rs4444235 and rs1957636) and 20p12.3 (rs961253 and rs4813802). These markers are located close to two of the genes involved in bone morphogenetic protein (BMP) signaling (BMP4 and BMP2, respectively). By investigating these four SNPs in an initial cohort of Spanish origin, we found substantial evidence that minor allele frequencies (MAFs) may be different in northern and southern European populations. Therefore, we genotyped three additional southern European cohorts comprising a total of 2028 cases and 4273 controls. The meta-analysis results show that only one of the association signals (rs961253) is effectively replicated in the southern European populations, despite adequate power to detect all four. The other three SNPs (rs4444235, rs1957636 and rs4813802) presented discordant results in MAFs and linkage disequilibrium patterns between northern and southern European cohorts. We hypothesize that this lack of replication could be the result of differential tagging of the functional variant in both sets of populations. Were this true, it would have complex consequences in both our ability to understand the nature of the real causative variants, as well as for further study designs.


Towards an understanding of inequalities in accessing residential and nursing home provision: The role of geographical approaches.
Suggestions of the existence of so-called 'social care deserts' in England in the years leading up to the COVID-19 pandemic drew attention to the potential impact of geographical inequalities on the availability of residential, nursing and domiciliary care. To date, much of this analysis has been conducted at spatially aggregated scales such as that of local authorities or postcode sector. Hidden within such aggregate-level analysis however are geographical differences in the local provision of care services. In this paper, we draw attention to geographical modelling techniques that can be used to examine local trends in the supply of social care services in relation to potential demand. These spatial models can be used to examine variations in the number of facilities (or choice) within reasonable drive times/distances. Drawing on a national database of residential and nursing care beds in Wales for March 2020, we illustrate the potential of such techniques to provide an insight into current patterns in access to care homes, and to monitor future changes in the fall-out from the effects of the COVID-19 pandemic on the care home sector. The concentration of care home sites in metropolitan areas and in the heavily populated post-industrial valleys in the south-east is identified, but significant demand present in these areas ameliorates scores towards mid-range ratios. We conclude by suggesting that the types of techniques used in this study enable disparities in provision within localised areas to be better explored, thereby helping planners and policy makers to address potential inequalities in provision.


UV-screening and springtime recovery of photosynthetic capacity in leaves of Vaccinium vitis-idaea above and below the snow pack.
Evergreen plants in boreal biomes undergo seasonal hardening and dehardening adjusting their photosynthetic capacity and photoprotection; acclimating to seasonal changes in temperature and irradiance. Leaf epidermal ultraviolet (UV)-screening by flavonols responds to solar radiation, perceived in part through increased ultraviolet-B (UV-B) radiation, and is a candidate trait to provide cross-photoprotection. At Hyytiälä Forestry Station, central Finland, we examined whether the accumulation of flavonols was higher in leaves of Vaccinium vitis-idaea L. growing above the snowpack compared with those below the snowpack. We found that leaves exposed to colder temperatures and higher solar radiation towards the top of hummocks suffered greater photoinhibition than those at the base of hummocks. Epidermal UV-screening was highest in upper-hummock leaves, particularly during winter when lower leaves were beneath the snowpack. There was also a negative relationship between indices of flavonols and anthocyanins across all leaves suggesting fine-tuning of flavonoid composition for screening vs. antioxidant activity in response to temperature and irradiance. However, the positive correlation between the maximum quantum yield of photosystem II photochemistry (Fv/Fm) and flavonol accumulation in upper hummock leaves during dehardening did not confer on them any greater cross-protection than would be expected from the general relationship of Fv/Fm with temperature and irradiance (throughout the hummocks). Irrespective of timing of snow-melt, photosynthesis fully recovered in all leaves, suggesting that V. vitis-idaea has the potential to exploit the continuing trend for longer growing seasons in central Finland without incurring significant impairment from reduced duration of snow cover.


Effects of intravenous zoledronic acid plus subcutaneous teriparatide [rhPTH(1-34)] in postmenopausal osteoporosis.
Clinical data suggest concomitant therapy with bisphosphonates and parathyroid hormone (PTH) may blunt the anabolic effect of PTH; rodent models suggest that infrequently administered bisphosphonates may interact differently. To evaluate the effects of combination therapy with an intravenous infusion of zoledronic acid 5 mg and daily subcutaneous recombinant human (rh)PTH(1-34) (teriparatide) 20 µg versus either agent alone on bone mineral density (BMD) and bone turnover markers, we conducted a 1-year multicenter, multinational, randomized, partial double-blinded, controlled trial. 412 postmenopausal women with osteoporosis (mean age 65 ± 9 years) were randomized to a single infusion of zoledronic acid 5 mg plus daily subcutaneous teriparatide 20 µg (n = 137), zoledronic acid alone (n = 137), or teriparatide alone (n = 138). The primary endpoint was percentage increase in lumbar spine BMD (assessed by dual-energy X-ray absorptiometry [DXA]) at 52 weeks versus baseline. Secondary endpoints included change in BMD at the spine at earlier time points and at the total hip, trochanter, and femoral neck at all time points. At week 52, lumbar spine BMD had increased 7.5%, 7.0%, and 4.4% in the combination, teriparatide, and zoledronic acid groups, respectively (p < .001 for combination and teriparatide versus zoledronic acid). In the combination group, spine BMD increased more rapidly than with either agent alone (p < .001 versus both teriparatide and zoledronic acid at 13 and 26 weeks). Combination therapy increased total-hip BMD more than teriparatide alone at all times (all p < .01) and more than zoledronic acid at 13 weeks (p < .05), with final 52-week increments of 2.3%, 1.1%, and 2.2% in the combination, teriparatide, and zoledronic acid groups, respectively. With combination therapy, bone formation (assessed by serum N-terminal propeptide of type I collagen [PINP]) increased from 0 to 4 weeks, declined minimally from 4 to 8 weeks, and then rose throughout the trial, with levels above baseline from 6 to 12 months. Bone resorption (assessed by serum β-C-telopeptide of type I collagen [β-CTX]) was markedly reduced with combination therapy from 0 to 8 weeks (a reduction of similar magnitude to that seen with zoledronic acid alone), followed by a gradual increase after week 8, with levels remaining above baseline for the latter half of the year. Levels for both markers were significantly lower with combination therapy versus teriparatide alone (p < .002). Limitations of the study included its short duration, lack of endpoints beyond DXA-based BMD (e.g., quantitative computed tomography and finite-element modeling for bone strength), lack of teriparatide placebo, and insufficient power for fracture outcomes. We conclude that while teriparatide increases spine BMD more than zoledronic acid and zoledronic acid increases hip BMD more than teriparatide, combination therapy provides the largest, most rapid increments when both spine and hip sites are considered.


Frequency and predictors of individual treatment outcomes (response, remission, exacerbation, and relapse) in clinical adolescents with nonsuicidal self-injury.
Nonsuicidal self-injury (NSSI) is prevalent in adolescent clinical samples. There is evidence that NSSI can be treated effectively but data on individual treatment outcomes is limited. The goal of this study was to examine response, remission, exacerbation, and relapse rates over one and two years, respectively, among a clinical sample of adolescents with NSSI. Furthermore, we aimed to identify clinically relevant predictors of NSSI trajectories. The sample consists of n = 203 adolescents (12-17 y., 94% female) from a specialized outpatient clinic for risk-taking and self-harming behavior with NSSI on at least five days in the six months before first assessment. Assessments were completed at baseline and one (FU1) and two (FU2) years later using structured clinical interviews and self-report questionnaires. At FU1, 75% reported a reduction in NSSI frequency by at least 50% (treatment response); among those, one third (25% of the entire sample) achieved a remission (0 NSSI); an exacerbation (⩾50% more NSSI) was observed in 11% of patients. Of those in remission, 41% relapsed one year later. Predictors of non-response or non-remission were inpatient treatment and depressive symptoms. Adolescents with lower NSSI frequency at baseline had a higher risk of exacerbation. Due to limited sample size at FU2 no prediction model for relapse was established. While most adolescents presenting with NSSI achieved significant improvement, more attention should be paid to the rather low rates of full remission. Prediction and early detection of individuals who deteriorate during or relapse after treatment is critical.


Targeting αvβ3 and αvβ5 integrins inhibits pulmonary metastasis in an intratibial xenograft osteosarcoma mouse model.
Osteosarcoma is an aggressive bone cancer that has a high propensity for metastasis to the lungs. Patients with metastatic disease face a very poor prognosis. Therefore, novel therapeutics, efficiently suppressing the metastatic process, are urgently needed. Integrins play a pivotal role in tumor cell adhesion, motility and metastasis. Here, we evaluated αvβ3 and αvβ5 integrin inhibition with cilengitide as a novel metastasis-suppressive therapeutic approach in osteosarcoma. Immunohistochemical analysis of αvβ3 and αvβ5 integrins expression in a tissue microarray of tumor specimens collected from osteosarcoma patients revealed that αvβ5 integrin is mainly found on tumor cells, whereas αvβ3 is predominantly expressed by stromal cells. In vitro functional assays demonstrated that cilengitide dose-dependently inhibited de novo adhesion, provoked detachment and inhibited migration of osteosarcoma cell lines. Cilengitide induced a decline in cell viability, blocked the cell cycle in the G1 phase and caused anoikis by activation of the Hippo pathway. In a xenograft orthotopic mouse model cilengitide minimally affected intratibial primary tumor growth but, importantly, suppressed pulmonary metastasis. The data demonstrate that targeting αvβ3 and αvβ5 integrins in osteosarcoma should be considered as a novel therapeutic option for patients with metastatic disease.


The chemistry of silica and its potential health benefits.
There is considerable interest in the effects of silica on human health in contrast to prior research which focused solely on the toxic effects of inhaled crystalline silica. However, multiple forms of silica exist in nature and silicon, a component, is the second most prevalent element after oxygen. Silica has widespread industrial applications including use as a food additive, i.e., anti-caking agent, as a means to clarify beverages, control viscosity, as an anti-foaming agent, dough modifier, and as an excipient in drugs and vitamins. Chemically, silica is an oxide of silicon, viz., silicon dioxide, and is generally colorless to white and insoluble in water. When associated with metals or minerals the family of silicates is formed. There are several water soluble forms of silica referred collectively to as silicic acid (ortho, meta, di, and tri-silicates), which are present in surface and well water in the range of 1--100 mg/L. Orthosilicic acid is the form predominantly absorbed by humans and is found in numerous tissues including bone, tendons, aorta, liver and kidney. Compelling data suggest that silica is essential for health although no RDI has been established. However, deficiency induces deformities in skull and peripheral bones, poorly formed joints, reduced contents of cartilage, collagen, and disruption of mineral balance in the femur and vertebrae. Very little toxicity data exist regarding aqueous silica consumption due, in part, to the lack of anecdotal reports of toxicity and general presumption of safety. However, a few rodent studies have been conducted, which indicate a No Observed Adverse Effects Level (NOAEL) of 50,000 ppm (mg/L) for dietary silica. In conclusion, many forms of silica exist in nature and compelling data support myriad beneficial effects of silica in water.


Optimal dispersion maps for wavelength-division-multiplexed soliton transmission.
We propose an advantageous dispersion-management scheme for wavelength-division-multiplexed soliton transmission in which optimal launch points are obtained whose locations are independent of the fibers' dispersion parameters. Inasmuch as using such optimal launch points minimizes dispersively shed radiation, it is possible to optimize the transmission simultaneously in several different channels. For the particular case of a two-step dispersion map we demonstrate that this result can be achieved by proper choice of the fiber lengths.


Visit-to-visit (long-term) and ambulatory (short-term) blood pressure variability to predict mortality in an elderly hypertensive population.
To explore the association of different types of blood pressure (BP) variability measures estimated from either short-term ambulatory reading-to-reading or long-term clinic visit-to-visit BP records with long-term survival in an elderly treated hypertensive population. A subset of patients (n = 508) aged at least 65-years was studied from the Second Australian National Blood Pressure study. We estimated SBP and DBP BP variability as the SD of ambulatory (24-h, daytime, night-time) and clinic visit-to-visit BP directly from all corresponding on-treatment within-individual BP records. Ambulatory 'weighted day-night' variability was calculated as a weighted mean of daytime and night-time SD. Cox-proportional hazard models adjusted for baseline risk factors (Model 1) and corresponding on-treatment BP (Model 2) or average night-time SBP (best predictive BP measure for outcome) (Model 3) were used to determine the relationship between long-term outcome and BP variability. Over a median of 10.6 years, 101 patients died from any cause, of which 51 deaths were cardiovascular. We observed increase in 'daytime' and 'weighted day-night' SBP/DBP variability was significantly associated with increased all-cause mortality in all models. For cardiovascular mortality, only 'weighted day-night' SBP variability significantly predicted risk in all models (Model 3 hazard ratio: 1.09, 95% confidence interval: 1.00-1.19, P = 0.04). Long-term BP variability was not associated with any outcome. On direct comparison, both 'daytime' and 'weighted day-night' BP variability measures provided similar prognostic information. Short-term 'daytime' and 'weighted day-night' SBP variability from ambulatory BP recordings was a better predictor of mortality in elderly treated hypertensive patients than long-term BP variability from visit-to-visit BP recordings.


Neuroprotective effects of polysaccharides from wolfberry, the fruits of Lycium barbarum, against homocysteine-induced toxicity in rat cortical neurons.
Previous clinical and epidemiological studies have suggested that elevated plasma homocysteine (Hcy) levels increased the risk of Alzheime's disease (AD). Although the underlying mechanisms of its toxicity are elusive, it has been shown that Hcy damages neurons by inducing apoptosis, DNA fragmentation, and tau hyperphosphorylation. Wolfberry (Lycium barbarum) is a fruit that is known for its eye-protective and anti-aging properties in Asian countries. Previous studies from our laboratory have demonstrated that polysaccharides derived from wolfberry (LBA) have the ability to protect neurons from amyloid-beta (Abeta) peptide neurotoxicity. We hypothesize that the neuroprotective effects of wolfberry is not limited to Abeta and can also provide protection against other AD risk factors. In this study, we aim to elucidate the neuroprotective effects of wolfberry against Hcy-induced neuronal damage. Our data showed that LBA treatment significantly attenuated Hcy-induced neuronal cell death and apoptosis in primary cortical neurons as demonstrated by LDH and caspase-3 like activity assay. LBA also significantly reduced Hcy-induced tau phosphorylation at tau-1 (Ser198/199/202), pS396 (Ser396), and pS214 (Ser214) epitopes as well as cleavage of tau. At the same time, we also found that the phosphorylation level of p-GSK3beta (Ser9/Tyr 216) remained unchanged among different treatment groups at all detected time points. LBA treatment suppressed elevation of both p-ERK and p-JNK. In summary, our data demonstrated that LBA exerted neuroprotective effects on cortical neurons exposed to Hcy. Therefore, LBA has the potential to be a diseasemodifying agent for the prevention of AD.


Inheritance and identification of molecular markers associated with a novel dwarfing gene in barley.
Dwarfing genes have widely been used in barley breeding program. More than 30 types of dwarfs or semidwarfs have been reported, but a few has been exploited in barley breeding because pleiotropic effects of dwarfing genes cause some undesired traits. The plant architecture of newly discovered dwarfing germplasm "Huaai 11" consisted of desirable agronomic traits such as shortened stature and early maturity. Genetic factor controlling the plant height in dwarf line Huaai 11 was investigated. The Huaai 11 was crossed with tall varieties Monker, Mpyt, Zhenongda 3, Zaoshu 3, Advance, Huadamai 1, Huadamai 6, Hyproly and Ris01508. All the F1 plants displayed tall trait. Both tall and dwarf plants appeared in all the F2 populations with a 3:1 segregation ratio, suggesting that dwarfism of Huaai 11 is controlled by a single recessive gene, btwd1. Allelism test indicated that this dwarfing gene in the Huaai 11 is nonallelic with the gene br, uzu, sdw1 and denso. Using a double haploid population derived from a cross of Huadamai 6 and Huaai 11 and SSR markers the novel dwarfing gene was mapped onto the long arm of chromosome 7H, and closely linked to Bmac031 and Bmac167 with genetic distance of 2.2 cM. Huaai 11 is a new source of dwarf for broadening the genetic base of dwarfism. This dwarf source was controlled by a recessive dwarfing gene btwd1, was mapped onto the long arm of chromosome 7H.


Methylation profiling of ductal carcinoma in situ and its relationship to histopathological features.
DNA methylation is a well-studied biomarker in invasive breast cancer, but its role in ductal carcinoma in situ (DCIS) is less well characterized. The aims of this study are to assess the methylation profile in DCIS for a panel of well-characterized genes that are frequently methylated in breast cancer, to investigate the relationship of methylation with pathological features, and to perform a proof-of-principle study to evaluate the practicality of methylation as a biomarker in diagnostic DCIS material. Promoter CpG island methylation for a panel of 11 breast cancer-related genes was performed by methylation-sensitive high resolution melting (MS-HRM). Formalin-fixed, paraffin-embedded (FFPE) biopsies from 72 samples of pure DCIS (DCIS occurring in the absence of synchronous invasive carcinoma), 10 samples of mixed DCIS (DCIS adjacent to invasive carcinoma), and 18 samples of normal breast epithelium adjacent to a DCIS lesion were micro-dissected prior to DNA extraction. Methylation was seen for all the tested genes except BRCA1. RASSF1A was the most frequently methylated gene (90% of DCIS samples) and its methylation was associated with comedo necrosis (p = 0.018). Cluster analysis based on the methylation profile revealed four groups, the highly methylated cluster being significantly associated with high nuclear grade, HER2 amplification, negative estrogen receptor (ER) α status, and negative progesterone receptor (PgR) status, (p = 0.038, p = 0.018, p <0.001, p = 0.001, respectively). Methylation of APC (p = 0.017), CDH13 (p = 0.017), and RARβ (p <0.001) was associated with negative ERα status. Methylation of CDH13 (p <0.001), and RARβ (p = 0.001) was associated with negative PgR status. Methylation of APC (p = 0.013) and CDH13 (p = 0.026) was associated with high nuclear grade. Methylation of CDH13 (p = 0.009), and RARβ (p = 0.042) was associated with HER2-amplification. DNA methylation can be assessed in FFPE-derived samples using suitable methodologies. Methylation of a panel of genes that are known to be methylated in invasive breast cancer was able to classify DCIS into distinct groups and was differentially associated with phenotypic features in DCIS.


Spontaneous development of autoimmune uveoretinitis in nude mice following reconstitution with embryonic rat thymus.
This paper describes the spontaneous occurrence of an autoimmune uveoretinitis in nude (nu/nu) mice reconstituted when 4 weeks old by the grafting of rat embryonic thymus. The uveoretinitis was characterized histologically by progressive loss of the photoreceptor layer, observed in 4.0, 17.6, 42.9% and 71.4% of such mice at 3, 5, 7 and 12 months of age, respectively. Mice with uveoretinitis were shown to have serum IgG antibody reactive by indirect immunofluorescence with retinal photoreceptors, and with interphotoreceptor retinoid-binding protein (IRBP), but not retinal S-antigen, by immunoblotting and ELISA. A uveoretinitis could be adoptively transferred to syngeneic ungrafted nude mice by splenic CD4+ T cells from diseased animals. This is the first experimental model of a (T cell mediated) autoimmune uveoretinitis which develops spontaneously and which is not dependent upon deliberate sensitization with retinal antigens and adjuvants.


Legalizing and Regulating Marijuana in Canada: Review of Potential Economic, Social, and Health Impacts.
Notwithstanding a century of prohibition, marijuana is the most widely used illicit substance in Canada. Due to the growing public acceptance of recreational marijuana use and ineffectiveness of the existing control system in Canada, the issue surrounding legalizing this illicit drug has received considerable public and political attentions in recent years. Consequently, the newly elected Liberal Government has formally announced that Canada will introduce legislation in the spring of 2017 to start legalizing and regulating marijuana. This editorial aims to provide a brief overview on potential economic, social, and public health impacts of legal marijuana in Canada. The legalization could increase tax revenue through the taxation levied on marijuana products and could also allow the Government to save citizens' tax dollars currently being spent on prohibition enforcement. Moreover, legalization could also remove the criminal element from marijuana market and reduce the size of Canada's black market and its consequences for the society. Nevertheless, it may also lead to some public health problems, including increasing in the uptake of the drug, accidents and injuries. The legalization should be accompanied with comprehensive strategies to keep the drug out of the hands of minors while increasing awareness and knowledge on harmful effects of the drug. In order to get better insights on how to develop an appropriate framework to legalize marijuana, Canada should closely watch the development in the neighboring country, the United States, where some of its states viz, Colorado, Oregon, Washington, and Alaska have already legalized recreational use of marijuana.


Genome-level transcription data of Yersinia pestis analyzed with a new metabolic constraint-based approach.
Constraint-based computational approaches, such as flux balance analysis (FBA), have proven successful in modeling genome-level metabolic behavior for conditions where a set of simple cellular objectives can be clearly articulated. Recently, the necessity to expand the current range of constraint-based methods to incorporate high-throughput experimental data has been acknowledged by the proposal of several methods. However, these methods have rarely been used to address cellular metabolic responses to some relevant perturbations such as antimicrobial or temperature-induced stress. Here, we present a new method for combining gene-expression data with FBA (GX-FBA) that allows modeling of genome-level metabolic response to a broad range of environmental perturbations within a constraint-based framework. The method uses mRNA expression data to guide hierarchical regulation of cellular metabolism subject to the interconnectivity of the metabolic network. We applied GX-FBA to a genome-scale model of metabolism in the gram negative bacterium Yersinia pestis and analyzed its metabolic response to (i) variations in temperature known to induce virulence, and (ii) antibiotic stress. Without imposition of any a priori behavioral constraints, our results show strong agreement with reported phenotypes. Our analyses also lead to novel insights into how Y. pestis uses metabolic adjustments to counter different forms of stress. Comparisons of GX-FBA predicted metabolic states with fluxomic measurements and different reported post-stress phenotypes suggest that mass conservation constraints and network connectivity can be an effective representative of metabolic flux regulation in constraint-based models. We believe that our approach will be of aid in the in silico evaluation of cellular goals under different conditions and can be used for a variety of analyses such as identification of potential drug targets and their action.


ANF-mediated renal cGMP generation in congestive heart failure.
Previous studies have demonstrated that the biological actions of atrial natriuretic factor (ANF) are mediated via increases in its intracellular second messenger guanosine 3',5'-cyclic monophosphate (cGMP). Because cGMP egresses rapidly from target cells after ANF binding to particulate guanylate cyclase-linked receptors, extracellular cGMP may be a useful biological marker for the action of ANF in vivo under pathophysiological conditions. The present studies tested the hypothesis that the avid sodium retention and renal ANF resistance characteristic of chronic congestive heart failure (CHF) are associated with attenuated renal cGMP responses to ANF. We assessed the natriuretic and cGMP responses to endogenous and exogenous ANF during the evolution of CHF produced by 6 days of rapid ventricular pacing in conscious dogs (n = 6). Simultaneous measurement of plasma and urinary cGMP concentrations allowed determination of the net renal generation of cGMP, an indicator of the renal contribution to total urinary cGMP excretion. In early CHF, increased sodium excretion and renal cGMP production were observed in association with increases in plasma ANF. Exogenous ANF administration (10 micrograms/kg iv) before CHF also produced parallel increases in sodium excretion and renal cGMP production. In more advanced CHF produced by 6 days of pacing, we observed avid sodium retention in association with reversal of earlier increases in renal cGMP production despite progressive increases in circulating ANF. Natriuretic and renal cGMP responses to exogenous ANF were similarly attenuated in chronic CHF. These studies suggest that 1) renal cGMP production is a useful biological marker for the renal natriuretic action of ANF; 2) endogenous ANF contributes to the maintenance of sodium excretion in early CHF via increases in renal cGMP production; and 3) the avid sodium retention and renal ANF resistance in advanced CHF are, in part, linked to attenuated renal cGMP responses to endogenous and exogenous ANF.


Effect of endocardial trabeculae on left ventricular measurements and measurement reproducibility at cardiovascular MR imaging.
To prospectively assess the effect of including or excluding endocardial trabeculae in left ventricular (LV) measurements and the reproducibility of these measurements at cine cardiovascular magnetic resonance (MR) imaging with true fast imaging with steady-state precession (FISP). The study was approved by the local ethics committee, and each subject gave informed consent before participating. Twenty healthy subjects and 20 consecutive patients underwent 1.5-T cardiovascular MR imaging. Seven to 12 short-axis views encompassing the entire LV were acquired by using true FISP. Endocardial and epicardial contours were traced manually. The data sets in each patient were analyzed twice: with inclusion of endocardial trabeculae in the LV cavity volume and with exclusion of endocardial trabeculae from the cavity volume. On the basis of these two contour sets, the end-diastolic (ED) and end-systolic (ES) LV volumes, ejection fraction (EF), and LV mass were calculated. Additionally, interobserver and interexamination reproducibility was assessed by using Bland-Altman analysis. Compared with exclusion of trabeculae, inclusion of trabeculae in the LV cavity volume resulted in significantly larger ED and ES LV volumes (mean differences, 21 mL +/- 11 [standard deviation] and 19 mL +/- 33, respectively; P < .001) and lower EFs (mean difference, -2% +/- 2; P < .001). The calculated LV mass was significantly smaller with inclusion than with exclusion of trabeculae (mean difference, -21 g +/- 12; P < .001). All interobserver and interexamination limits of agreement based on inclusion of trabeculae, except those for EF measurements, were superior to those based on exclusion of trabeculae. At measurement reproducibility comparisons, differences in interobserver ED LV volume and LV mass and interexamination LV mass were statistically significant, favoring the inclusion of trabeculae in the LV cavity volume. Trabeculae significantly affect quantifications of LV volume and mass. The superior reproducibility of LV measurements with the inclusion of endocardial trabeculae in the cavity volume favors this tracing algorithm for clinical use.