Genome mining and metabolic profiling of the rhizosphere bacterium Pseudomonas sp. SH-C52 for antimicrobial compounds.
The plant microbiome represents an enormous untapped resource for discovering novel genes and bioactive compounds. Previously, we isolated Pseudomonas sp. SH-C52 from the rhizosphere of sugar beet plants grown in a soil suppressive to the fungal pathogen Rhizoctonia solani and showed that its antifungal activity is, in part, attributed to the production of the chlorinated 9-amino-acid lipopeptide thanamycin (Mendes et al., 2011). To get more insight into its biosynthetic repertoire, the genome of Pseudomonas sp. SH-C52 was sequenced and subjected to in silico, mutational and functional analyses. The sequencing revealed a genome size of 6.3 Mb and 5579 predicted ORFs. Phylogenetic analysis placed strain SH-C52 within the Pseudomonas corrugata clade. In silico analysis for secondary metabolites revealed a total of six non-ribosomal peptide synthetase (NRPS) gene clusters, including the two previously described NRPS clusters for thanamycin and the 2-amino acid antibacterial lipopeptide brabantamide. Here we show that thanamycin also has activity against an array of other fungi and that brabantamide A exhibits anti-oomycete activity and affects phospholipases of the late blight pathogen Phytophthora infestans. Most notably, mass spectrometry led to the discovery of a third lipopeptide, designated thanapeptin, with a 22-amino-acid peptide moiety. Seven structural variants of thanapeptin were found with varying degrees of activity against P. infestans. Of the remaining four NRPS clusters, one was predicted to encode for yet another and unknown lipopeptide with a predicted peptide moiety of 8-amino acids. Collectively, these results show an enormous metabolic potential for Pseudomonas sp. SH-C52, with at least three structurally diverse lipopeptides, each with a different antimicrobial activity spectrum.


A novel cell surface antigen, immature thymocyte antigen-1, is involved in the differentiation of murine thymocytes.
A mAb, 1-23, which recognizes a novel cell surface Ag on immature murine thymocytes (designated as IMT-1 for immature thymocyte antigen-1) was prepared. IMT-1 was found to be expressed on 40 to 50% of CD4-8- double negative (DN), 5 to 10% of CD4-8+, and 5 to 20% of CD4+8+ double positive (DP) thymocytes in adult mice, but not expressed on CD4+8- thymocytes or peripheral T lymphocytes. It was not expressed on either nonlymphoid cell lines or thymic stromal cells. In subsets of DN thymocytes, IMT-1 was detected on 40 to 50% of the heat-stable Ag+, 15% of CD44+25+, 70% of CD44-25+, and 70% of CD44-25- populations, whereas it was not detected on heat-stable Ag- and CD44+25- populations. IMT-1+ thymocytes expressed low levels of or no CD3 molecules. In fetal thymuses, IMT-1 was expressed on a minor population of thymocytes at day 14.5 and 15.5 of gestation. However, at day 16.5 of gestation, a majority of DN as well as DP thymocytes became IMT-1-positive. Addition of the 1-23 Ab to the fetal thymus organ culture relatively increased CD8+ SP thymocytes. These results show that IMT-1 is expressed during the late DN stage as well as the early DP stage of thymocyte maturation and suggest the possible involvement of IMT-1 in thymocyte differentiation.


Adiabatic theorem and continuous coupled-mode theory for efficient taper transitions in photonic crystals.
We prove that an adiabatic theorem generally holds for slow tapers in photonic crystals and other strongly grated waveguides with arbitrary index modulation, exactly as in conventional waveguides. This provides a guaranteed pathway to efficient and broad-bandwidth couplers with, e.g., uniform waveguides. We show that adiabatic transmission can only occur, however, if the operating mode is propagating (nonevanescent) and guided at every point in the taper. Moreover, we demonstrate how straightforward taper designs in photonic crystals can violate these conditions, but that adiabaticity is restored by simple design principles involving only the independent band structures of the intermediate gratings. For these and other analyses, we develop a generalization of the standard coupled-mode theory to handle arbitrary nonuniform gratings via an instantaneous Bloch-mode basis, yielding a continuous set of differential equations for the basis coefficients. We show how one can thereby compute semianalytical reflection and transmission through crystal tapers of almost any length, using only a single pair of modes in the unit cells of uniform gratings. Unlike other numerical methods, our technique becomes more accurate as the taper becomes more gradual, with no significant increase in the computation time or memory. We also include numerical examples comparing to a well-established scattering-matrix method in two dimensions.


Why the NIH Trial to Assess Chelation Therapy (TACT) should be abandoned.
The National Institutes of Health (NIH) Trial to Assess Chelation Therapy (TACT) was begun in 2003 and is expected to be completed in 2009. It is a trial of office-based, intravenous disodium ethylene-diamine-tetra-acetic acid (Na(2)EDTA) as a treatment for coronary artery disease (CAD). A few case series in the 1950s and early 1960s had found Na(2)EDTA to be ineffective for CAD or peripheral vascular disease (PVD). Nevertheless, a few hundred physicians, almost all of whom advocate other dubious treatments, continued to peddle chelation as an office treatment. They claim that chelation dramatically improves symptoms and prolongs life in 80% to 90% of patients. In response, academics performed 4 controlled trials during the 1990s. None favored chelation, but chelationists repudiated those findings. We have investigated the method and the trial. We present our findings in 4 parts: history, origin and nature of the TACT, state of the evidence, and risks. We present evidence that chelationists and their organization, the American College for Advancement in Medicine, used political connections to pressure the NIH to fund the TACT. The TACT protocols justified the trial by misrepresenting case series and by ignoring evidence of risks. The trial employs nearly 100 unfit co-investigators. It conflates disodium EDTA and another, somewhat safer drug. It lacks precautions necessary to minimize risks. The consent form reflects those shortcomings and fails to disclose apparent proprietary interests. The trial's outcome will be unreliable and almost certainly equivocal, thus defeating its stated purpose. We conclude that the TACT is unethical, dangerous, pointless, and wasteful. It should be abandoned.


Effect of comorbidity on quality of life and treatment selection in patients with squamous cell carcinoma of the head and neck.
Comorbidity is significantly associated with diminished survival and quality of life (QOL) after treatment of head and neck squamous cell carcinoma (HNSCC). We sought to determine whether comorbidity influenced pretreatment QOL scores and treatment selection in patients with HNSCC. The medical records of all patients diagnosed with HNSCC who participated in pretreatment QOL analysis over a 15-month period were retrospectively reviewed. Patients with a history of prior treatment for head and neck cancer, unresectable, or distant metastatic disease were excluded. The University of Washington (UW) QOL questionnaire, Performance Status Scale (PSS), and Karnofsky score were used to measure pretreatment QOL. Comorbidity was graded using the Modified Medical Comorbidity Index. Of 75 patients who met study criteria, 33 underwent primary surgical therapy, and 42 underwent nonoperative (radiation or chemoradiation) therapy. Treatment groups did not differ with respect to patient demographics, UW QOL scores, PSS scores, Karnofsky score, or comorbidity. Treatment groups differed significantly by disease stage and primary site. Patients with advanced stage disease (III/IV) or oropharyngeal primary tumors were more likely to undergo nonoperative treatment, compared with patients with early stage disease (I/II) or oral cavity primary tumors (P < .005). No significant association was found between comorbidity and pretreatment QOL scores. Comorbidity was not significantly associated with treatment selection or pretreatment QOL scores in patients with HNSCC. Location of the primary tumor and disease stage were significantly associated with treatment selection. Further studies are required to determine the effect of comorbidity on patient and tumor responses to treatment.


In vivo monitoring of drought-induced embolism in Callitris rhomboidea trees reveals wide variation in branchlet vulnerability and high resistance to tissue death.
Damage to the plant water transport system through xylem cavitation is known to be a driver of plant death in drought conditions. However, a lack of techniques to continuously monitor xylem embolism in whole plants in vivo has hampered our ability to investigate both how this damage propagates and the possible mechanistic link between xylem damage and tissue death. Using optical and fluorescence sensors, we monitored drought-induced xylem embolism accumulation and photosynthetic damage in vivo throughout the canopy of a drought-resistant conifer, Callitris rhomboidea, during drought treatments of c. 1 month duration. We show that drought-induced damage to the xylem can be monitored in vivo in whole trees during extended periods of water stress. Under these conditions, vulnerability of the xylem to cavitation varied widely among branchlets, with photosynthetic damage only recorded once > 90% of the xylem was cavitated. The variation in branchlet vulnerability has important implications for understanding how trees like C. rhomboidea survive drought, and the high resistance of branchlets to tissue damage points to runaway cavitation as a likely driver of tissue death in C. rhomboidea branch tips.


Gene Function Analysis in the Ubiquitous Human Commensal and Pathogen Malassezia Genus.
The genus Malassezia includes 14 species that are found on the skin of humans and animals and are associated with a number of diseases. Recent genome sequencing projects have defined the gene content of all 14 species; however, to date, genetic manipulation has not been possible for any species within this genus. Here, we develop and then optimize molecular tools for the transformation of Malassezia furfur and Malassezia sympodialis using Agrobacterium tumefaciens delivery of transfer DNA (T-DNA) molecules. These T-DNAs can insert randomly into the genome. In the case of M. furfur, targeted gene replacements were also achieved via homologous recombination, enabling deletion of the ADE2 gene for purine biosynthesis and of the LAC2 gene predicted to be involved in melanin biosynthesis. Hence, the introduction of exogenous DNA and direct gene manipulation are feasible in Malassezia species. Species in the genus Malassezia are a defining component of the microbiome of the surface of mammals. They are also associated with a wide range of skin disease symptoms. Many species are difficult to culture in vitro, and although genome sequences are available for the species in this genus, it has not been possible to assess gene function to date. In this study, we pursued a series of possible transformation methods and identified one that allows the introduction of DNA into two species of Malassezia, including the ability to make targeted integrations into the genome such that genes can be deleted. This research opens a new direction in terms of now being able to analyze gene functions in this little understood genus. These tools will contribute to define the mechanisms that lead to the commensalism and pathogenicity in this group of obligate fungi that are predominant on the skin of mammals.


Bronchoalveolar lavage findings in patients with chronic nonproductive cough.
Mast cells and eosinophils may play a role in the pathophysiology of chronic cough in nonasthmatics. It is unknown, however, whether degranulation of these cells occurs in the airways of such patients. Thirty-five nonsmoking patients referred with a chronic nonproductive cough (mean cough duration 76.2 months) were evaluated using a comprehensive diagnostic protocol. Bronchoalveolar lavage (BAL) cell differentials and BAL histamine, tryptase and eosinophilic cationic protein (ECP) concentrations were determined. Ten nonsmoking healthy volunteers served as controls. Diagnostic subgroups were identified: eight postnasal drip syndrome (PNDS), seven cough variant asthma (CVA), seven gastro-esophageal reflux (GOR), seven dual aetiology and six idiopathic. Nonasthmatic coughers (NAC) were characterized as those patients without bronchial hyperresponsiveness on histamine challenge and whose cough had either responded to therapy for PNDS or GOR or failed to improve with antiasthma therapy. There was a significant increase in both eosinophil and mast cell numbers (p<0.05) and in histamine levels (p = 0.027) when NAC patients were compared with controls. Tryptase and ECP levels were elevated in 7 of 23 and 6 of 23 NAC patients, respectively. In conclusion, airway inflammatory cell numbers are not only increased but also activated, suggesting an important role for airways inflammation in the pathophysiology of chronic nonproductive cough.


Intrinsically Disordered Protein TEX264 Mediates ER-phagy.
Certain proteins and organelles can be selectively degraded by autophagy. Typical substrates and receptors of selective autophagy have LC3-interacting regions (LIRs) that bind to autophagosomal LC3 and GABARAP family proteins. Here, we performed a differential interactome screen using wild-type LC3B and a LIR recognition-deficient mutant and identified TEX264 as a receptor for autophagic degradation of the endoplasmic reticulum (ER-phagy). TEX264 is an ER protein with a single transmembrane domain and a LIR motif. TEX264 interacts with LC3 and GABARAP family proteins more efficiently and is expressed more ubiquitously than previously known ER-phagy receptors. ER-phagy is profoundly blocked by deletion of TEX264 alone and almost completely by additional deletion of FAM134B and CCPG1. A long intrinsically disordered region of TEX264 is required for its ER-phagy receptor function to bridge the gap between the ER and autophagosomal membranes independently of its amino acid sequence. These results suggest that TEX264 is a major ER-phagy receptor.


Translating diabetes prevention into native Hawaiian and Pacific Islander communities: the PILI 'Ohana Pilot project.
Native Hawaiians (NH) and Other Pacific Islanders (OPI) bear an excess burden of diabetes health disparities. Translation of empirically tested interventions such as the Diabetes Prevention Program Lifestyle Intervention (DPP-LI) offers the potential for reversing these trends. Yet, little is known about how best to translate efficacious interventions into public health practice, particularly among racial/ethnic minority populations. Community-based participatory research (CBPR) is an approach that engages the community in the research process and has recently been proposed as a means to improve the translation of research into community practice. To address diabetes health disparities in NHOPIs, CBPR approaches were used to: (1) culturally adapt the DPP-LI for NHOPI communities; and (2) implement and examine the effectiveness of the culturally-adapted program to promote weight loss in 5 NHOPI communities. Informant interviews (n=15) and focus groups (n=15, with 112 NHOPI participants) were completed to inform the cultural adaptation of the DPP-LI program. A team of 5 community investigators and 1 academic research team collaboratively developed and implemented the 12-week pilot study to assess the effectiveness of the culturally adapted program. A total of 127 NHOPIs participated in focus groups and informant interviews that resulted in the creation of a significantly modified version of the DPP-LI, entitled the PILI 'Ohana Lifestyle Intervention (POLI). In the pilot study, 239 NHOPIs were enrolled and after 12 weeks (post-program), mean weight loss was -1.5 kg (95%CI -2.0, -1.0) with 26% of participants losing > or = 3% of their baseline weight. Mean weight loss among participants who completed all 8 lessons at 12 weeks was significantly higher (-1.8 kg, 95%CI -2.3, -1.3) than participants who completed less than 8 lessons (-0.70 kg, 95%CI -1.1, -0.29). A fully engaged CBPR approach was successful in translating an evidence based diabetes prevention program into a culturally relevant intervention for NHOPI communities. This pilot study demonstrates that weight loss in high risk minority populations can be achieved over a short period of time using CBPR approaches.


First-line Immuno-Oncology Combination Therapies in Metastatic Renal-cell Carcinoma: Results from the International Metastatic Renal-cell Carcinoma Database Consortium.
In metastatic renal-cell carcinoma (mRCC), recent data have shown efficacy of first-line ipilimumab and nivolumab (ipi-nivo) as well as immuno-oncology (IO)/vascular endothelial growth factor (VEGF) inhibitor combinations. Comparative data between these strategies are limited. To compare the efficacy of ipi-nivo versus IO-VEGF (IOVE) combinations in mRCC, and describe practice patterns and effectiveness of second-line therapies. Using the International Metastatic Renal-cell Carcinoma Database Consortium (IMDC) dataset, patients treated with any first-line IOVE combination were compared with those treated with ipi-nivo. All patients received first-line IO combination therapies. First- and second-line response rates, time to treatment failure (TTF), time to next treatment (TNT), and overall survival (OS) were analysed. Hazard ratios were adjusted for IMDC risk factors. In total, 113 patients received IOVE combinations and 75 received ipi-nivo. For IOVE combinations versus ipi-nivo, first-line response rates were 33% versus 40% (between-group difference 7%, 95% confidence interval [CI] -8% to 22%, p = 0.4), TTF was 14.3 versus 10.2 mo (p = 0.2), TNT was 19.7 versus 17.9 mo (p = 0.4), and median OS was immature but not statistically different (p = 0.17). Adjusted hazard ratios for TTF, TNT, and OS were 0.71 (95% CI 0.46-1.12, p = 0.14), 0.65 (95% CI 0.38-1.11, p = 0.11), and 1.74 (95% CI 0.82-3.68, p = 0.14), respectively. Sixty-four (34%) patients received second-line treatment. In patients receiving subsequent VEGF-based therapy, second-line response rates were lower in the IOVE cohort than in the ipi-nivo cohort (15% vs 45%; between-group difference 30%, 95% CI 3-57%, p = 0.04; n = 40), though second-line TTF was not significantly different (3.7 vs 5.4 mo; p = 0.4; n = 55). Limitations include the study's retrospective design and sample size. There were no significant differences in first-line outcomes between IOVE combinations and ipi-nivo. Most patients received VEGF-based therapy in the second line. In this group, second-line response rate was greater in patients who received ipi-nivo initially. There were no significant differences in key first-line outcomes for patients with metastatic renal-cell carcinoma receiving immuno-oncology/vascular endothelial growth factor inhibitor combinations versus ipilimumab and nivolumab.


The history of neurosurgery at the University of Alabama at Birmingham.
: The Division of Neurosurgery at the University of Alabama at Birmingham was formally founded in 1954 under the leadership of James Garber Galbraith. The following 60 years would see neurosurgery at the forefront of the development of a nationally recognized medical center in the heart of Birmingham, Alabama. The Department of Neurosurgery now employs 14 faculty members, performs more than 4500 neurosurgical procedures annually, is active in clinical and laboratory research, and boasts a contemporary, comprehensive residency training program.


No prognostic value added by vitamin D pathway SNPs to current prognostic system for melanoma survival.
The prognostic improvement attributed to genetic markers over current prognostic system has not been well studied for melanoma. The goal of this study is to evaluate the added prognostic value of Vitamin D Pathway (VitD) SNPs to currently known clinical and demographic factors such as age, sex, Breslow thickness, mitosis and ulceration (CDF). We utilized two large independent well-characterized melanoma studies: the Genes, Environment, and Melanoma (GEM) and MD Anderson studies, and performed variable selection of VitD pathway SNPs and CDF using Random Survival Forest (RSF) method in addition to Cox proportional hazards models. The Harrell's C-index was used to compare the performance of model predictability. The population-based GEM study enrolled 3,578 incident cases of cutaneous melanoma (CM), and the hospital-based MD Anderson study consisted of 1,804 CM patients. Including both VitD SNPs and CDF yielded C-index of 0.85, which provided slight but not significant improvement by CDF alone (C-index = 0.83) in the GEM study. Similar results were observed in the independent MD Anderson study (C-index = 0.84 and 0.83, respectively). The Cox model identified no significant associations after adjusting for multiplicity. Our results do not support clinically significant prognostic improvements attributable to VitD pathway SNPs over current prognostic system for melanoma survival.


Experiencing community: perspectives of individuals diagnosed as having serious mental illness.
Community integration is recognized as a crucial component of recovery from serious mental illness. Although the construct of community integration can be measured with structured instruments, little is known about the subjective and experiential meaning of community and community involvement for persons with serious mental illness. In 2010, 30 individuals with serious mental illness treated in two public mental health clinics completed semistructured interviews that elicited the places and people that they associate with the experience of community and the larger meaning of community in their lives. Participants described four experiences as integral to their concepts of community: receiving help, minimizing risk, avoiding stigma, and giving back. Participants looked for communities that provide reliable support, and they described the need to manage community contact in order to protect themselves and others from their symptoms and from discrimination. Most participants experienced communities centered on mental health treatment or mentally ill peers as providing opportunities for positive engagement. The experience of having a serious mental illness shapes preferences for and perceptions of community in pervasive ways. Participants described community involvement not as a means to move away from illness experiences and identities but as a process that is substantially influenced by them. Mental health communities may help individuals with serious mental illness to both manage their illness and recognize and enjoy a sense of community. The findings indicate the need for further research on the relationship between community integration and outcome in serious mental illness.


Mindfulness Research Update: 2008.
OBJECTIVE: To briefly review the effects of mindfulness on the mind, the brain, the body, and behavior. METHODS: Selective review of MEDLINE, PsycINFO, and Google Scholar databases (2003-2008) using the terms "mindfulness", "meditation", "mental health", "physical health", "quality of life", and "stress reduction." A total of 52 exemplars of empirical and theoretical work were selected for review. RESULTS: Both basic and clinical research indicate that cultivating a more mindful way of being is associated with less emotional distress, more positive states of mind, and better quality of life. In addition, mindfulness practice can influence the brain, the autonomic nervous system, stress hormones, the immune system, and health behaviors, including eating, sleeping and substance use, in salutary ways. CONCLUSION: The application of cutting-edge technology toward understanding mindfulness - an "inner technology" - is elucidating new ways in which attention, awareness, acceptance, and compassion may promote optimal health - in mind, body, relationships, and spirit.


Deconstructing the mechanisms and consequences of TGF-β-induced EMT during cancer progression.
Transforming growth factor-β (TGF-β) is a potent pleiotropic cytokine that regulates mammalian development, differentiation, and homeostasis in essentially all cell types and tissues. TGF-β normally exerts anticancer activities by prohibiting cell proliferation and by creating cell microenvironments that inhibit cell motility, invasion, and metastasis. However, accumulating evidence indicates that the process of tumorigenesis, particularly that associated with metastatic progression, confers TGF-β with oncogenic activities, a functional switch known as the "TGF-β paradox." The molecular determinants governing the TGF-β paradox are complex and represent an intense area of investigation by researchers in academic and industrial settings. Recent findings link genetic and epigenetic events in mediating the acquisition of oncogenic activity by TGF-β, as do aberrant alterations within tumor microenvironments. These events coalesce to enable TGF-β to direct metastatic progression via the stimulation of epithelial-mesenchymal transition (EMT), which permits carcinoma cells to abandon polarized epithelial phenotypes in favor of apolar mesenchymal-like phenotypes. Attempts to deconstruct the EMT process induced by TGF-β have identified numerous signaling molecules, transcription factors, and microRNAs operant in mediating the initiation and resolution of this complex transdifferentiation event. In addition to its ability to enhance carcinoma cell invasion and metastasis, EMT also endows transitioned cells with stem-like properties, including the acquisition of self-renewal and tumor-initiating capabilities coupled to chemoresistance. Here, we review recent findings that delineate the pathophysiological mechanisms whereby EMT stimulated by TGF-β promotes metastatic progression and disease recurrence in human carcinomas.


A prospective descriptive study of cryptococcal meningitis in HIV uninfected patients in Vietnam - high prevalence of Cryptococcus neoformans var grubii in the absence of underlying disease.
Most cases of cryptococcal meningitis occur in patients with HIV infection: the course and outcome of disease in the apparently immunocompetent is much more poorly understood. We describe a cohort of HIV uninfected Vietnamese patients with cryptococcal meningitis in whom underlying disease is uncommon, and relate presenting features of patients and the characteristics of the infecting species to outcome. A prospective descriptive study of HIV negative patients with cryptococcal meningitis based at the Hospital for Tropical Diseases, Ho Chi Minh City. All patients had comprehensive clinical assessment at baseline, were cared for by a dedicated study team, and were followed up for 2 years. Clinical presentation was compared by infecting isolate and outcome. 57 patients were studied. Cryptococcus neoformans var grubii molecular type VN1 caused 70% of infections; C. gattii accounted for the rest. Most patients did not have underlying disease (81%), and the rate of underlying disease did not differ by infecting species. 11 patients died while in-patients (19.3%). Independent predictors of death were age > or = 60 years and a history of convulsions (odds ratios and 95% confidence intervals 8.7 (1 - 76), and 16.1 (1.6 - 161) respectively). Residual visual impairment was common, affecting 25 of 46 survivors (54.3%). Infecting species did not influence clinical phenotype or outcome. The minimum inhibitory concentrations of flucytosine and amphotericin B were significantly higher for C. neoformans var grubii compared with C. gattii (p < 0.001 and p = 0.01 respectively). In HIV uninfected individuals in Vietnam, cryptococcal meningitis occurs predominantly in people with no clear predisposing factor and is most commonly due to C. neoformans var grubii. The rates of mortality and visual loss are high and independent of infecting species. There are detectable differences in susceptibility to commonly used antifungal drugs between species, but the clinical significance of this is not clear.


Cameroon public health sector: shortage and inequalities in geographic distribution of health personnel.
Cameroon is classified by the World Health Organization (WHO) as having a critical shortage of health personnel. This is further complicated by the geographic distributional inequalities of the national health workforce. This shortfall impedes Cameroons' progress of improving the human resources for health (HRH) to meet up with the Millennium Development Goals (MDGs) by 2015. However, it is unknown whether the health workforce of Cameroon is distributed equally across geographic regions. Additionally, indicators other than population levels have not been used to measure health care needs. This study aimed to assess the adequacy, evenness of distribution and challenges faced by the health workforce across the different regions of Cameroon. National health personnel availability and distribution were assessed by use of end-of-year census data for 2011 obtained from the MoPH data base. The inequalities and distribution of the workforce were estimated using Gini coefficient and Lorenz curve and linear regression was used to determine the relation between health personnel density and selected health outcomes. Alternative indicators to determine health care needs were illustrated using concentration curves. Significant geographic inequalities in the availability of health workforce exist in Cameroon. Some regions have a higher number of physicians (per person) than others leading to poor health outcomes across the regions. 70 % of regions have a density of health personnel-to-population per 1,000 that is less than 1.5, implying acute shortage of health personnel. Poor working and living conditions, coupled with limited opportunities for career progress accounted for some documented 232 physicians and 205 nurses that migrated from the public sector. Significant distributional inequality was noticed when under-five infant mortality and malaria prevalence rate were used as indicators to measure health care needs. Our results show an absolute shortage of public health personnel in Cameroon that is further complicated by the geographic distributional inequalities across the regions of the nation. Cameroon aims to achieve universal health coverage by 2035; to realize this objective, policies targeting training, recruitment, retention and effective deployment of motivated and supported health workforce as well as the development and improvement of health infrastructures remain the major challenge.


Anticarcinogenic, cardioprotective, and other health benefits of tomato compounds lycopene, α-tomatine, and tomatidine in pure form and in fresh and processed tomatoes.
Tomatoes produce the bioactive compounds lycopene and α-tomatine that are reported to have potential health-promoting effects in animals and humans, but our understanding of the roles of these compounds in the diet is incomplete. Our current knowledge gained from the chemistry and analysis of these compounds in fresh and processed tomatoes and from studies on their bioavailability, bioactivity, and mechanisms of action against cancer cells and other beneficial bioactivities including antibiotic, anti-inflammatory, antioxidative, cardiovascular, and immunostimulating effects in cells, animals, and humans is discussed and interpreted here. Areas for future research are also suggested. The collated information and suggested research might contribute to a better understanding of the agronomical, biochemical, chemical, physiological, molecular, and cellular bases of the health-promoting effects and facilitate and guide further studies needed to optimize the use of lycopene and α-tomatine in pure form and in fresh tomatoes and processed tomato products to help prevent or treat human disease.


Oral diseases: a 14-year experience of a Chilean institution with a systematic review from eight countries.
Retrospective studies to assess the distribution of oral diseases (ODs) are helpful in estimating the prevalence of oral diagnoses in the population, and thus help in preventive and curative services. Prevalence and frequency data for ODs are available from many countries, but information from Chile is scarce. This study investigated the frequency of OD in a Chilean population. For this, we included all patients treated at the University of Talca (UTALCA, Chile) between 2001 and 2014. Patient characteristics were retrieved from medical files. To contextualize our results, we conducted a systematic review (SystRev) using Publish or Perish software (PoP), Google Scholar and MEDLINE/PubMed. One hundred sixty-six ODs were diagnosed, and the most prevalent groups were soft tissue tumours, epithelial pathology and salivary gland pathology. Individually, irritation fibroma, oral lichen planus (OLP) and mucocele were the most common diagnoses. ODs frequently affected unspecified parts of the mouth (including cheek, vestibule and retromolar area), gum, lips, tongue and palate. In the SystRev, the more studied diagnoses were leukoplakia, OLP and recurrent aphthous stomatitis; prevalent lesions included Fordyce's spots, recurrent aphthous stomatitis and fissured tongue. Chilean patients and SistRev shared almost all ODs. The results reflect ODs diagnosed in a specialized service of oral pathology and medicine in Chile and will allow the establishment of preventive/curative policies, adequate health services and dentistry curriculum.


Nikkomycin Z supersensitivity of an echinocandin-resistant mutant of Saccharomyces cerevisiae.
Echinocandins and nikkomycins are antibiotics that inhibit the synthesis of the essential cell wall polysaccharide polymers 1,3-beta-glucan and chitin, respectively. Some 40 echinocandin-resistant Saccharomyces cerevisiae mutants were isolated and assigned to five complementation groups. Four complementation groups contained mutants with 38 recessive mutations. The fifth complementation group comprised mutants with one dominant mutation, etg1-3 (strain MS10), and one semidominant mutation, etg1-4 (strain MS14). MS10 and MS14 are resistant to the semisynthetic pneumocandin B, L-733,560, and to aculeacin A but not to papulacandin. In addition, microsomal membranes of both mutant strains contain 1,3-beta-glucan synthase activity that is resistant to L-733,560 but not to papulacandin. Furthermore, MS14 is also supersensitive to nikkomycin Z. The echinocandin resistance and the nikkomycin Z supersensitivity of MS14 cosegregated in genetic crosses. The wild-type gene (designated ETG1 [C. Douglas, J. A. Marrinan, and M. B. Kurtz, J. Bacteriol. 176:5686-5696, 1994, and C. Douglas, F. Foor, J. A. Marrinan, N. Morin, J. B. Nielsen, A. Dahl, P. Mazur, W. Baginsky, W. Li, M. El-Sherbeini, J. A. Clemas, S. Mandala, B. R. Frommer, and M. B. Kurtz, Proc. Natl. Acad. Sci. USA, in press]) was isolated from a genomic library in the plasmid YCp50 by functional complementation of the nikkomycin Z supersensitivity phenotype. The cloned DNA also partially complements the echinocandin resistance phenotype, indicating that the two phenotypes are due to single mutations. The existence of a single mutation, in MS14, simultaneously affecting sensitivity to a glucan synthase inhibitor and a chitin synthase inhibitor implies a possible interaction between the two polymers at the cell surface.


Caseview: building the reference set.
There is a worldwide consensus for using the diagnosis related groups (DRG) when considering hospital activity. This tool leads to the production of tables of numbers (case mix), the interpretation of which is difficult. Therefore, methods aimed at facilitating this interpretation are needed. One of such methods is the case view, i.e. a graphical representation of the case mix. It reduces, in a way, each DRG to a "pixel", the set of the DRGs being an image (the case view). The reference set should be organized according to three criteria: medical/surgical, nosological and economic. This method can be used to answer theoretical questions or to visualize activity at the level of a hospital or at the level of a department. The purpose of this paper is to present important principles inherent in this graphic representation, both at the level of the method and at the level of the user.


The marine n-3 PUFA DHA evokes cytoprotection against oxidative stress and protein misfolding by inducing autophagy and NFE2L2 in human retinal pigment epithelial cells.
Accumulation and aggregation of misfolded proteins is a hallmark of several diseases collectively known as proteinopathies. Autophagy has a cytoprotective role in diseases associated with protein aggregates. Age-related macular degeneration (AMD) is the most common neurodegenerative eye disease that evokes blindness in elderly. AMD is characterized by degeneration of retinal pigment epithelial (RPE) cells and leads to loss of photoreceptor cells and central vision. The initial phase associates with accumulation of intracellular lipofuscin and extracellular deposits called drusen. Epidemiological studies have suggested an inverse correlation between dietary intake of marine n-3 polyunsaturated fatty acids (PUFAs) and the risk of developing neurodegenerative diseases, including AMD. However, the disease-preventive mechanism(s) mobilized by n-3 PUFAs is not completely understood. In human retinal pigment epithelial cells we find that physiologically relevant doses of the n-3 PUFA docosahexaenoic acid (DHA) induce a transient increase in cellular reactive oxygen species (ROS) levels that activates the oxidative stress response regulator NFE2L2/NRF2 (nuclear factor, erythroid derived 2, like 2). Simultaneously, there is a transient increase in intracellular protein aggregates containing SQSTM1/p62 (sequestosome 1) and an increase in autophagy. Pretreatment with DHA rescues the cells from cell cycle arrest induced by misfolded proteins or oxidative stress. Cells with a downregulated oxidative stress response, or autophagy, respond with reduced cell growth and survival after DHA supplementation. These results suggest that DHA both induces endogenous antioxidants and mobilizes selective autophagy of misfolded proteins. Both mechanisms could be relevant to reduce the risk of developing aggregate-associate diseases such as AMD.


Self-perceptions from people with Down syndrome.
This study asks people with Down syndrome (DS), ages 12 and older, about their self-perception so that their information could be shared with new and expectant parents of children with DS. We analyzed valid and reliable survey instruments from 284 people with DS on the mailing lists of 6 non-profit DS organizations around the country. Among those surveyed, nearly 99% of people with DS indicated that they were happy with their lives, 97% liked who they are, and 96% liked how they look. Nearly 99% people with DS expressed love for their families, and 97% liked their brothers and sisters. While 86% of people with DS felt they could make friends easily, those with difficulties mostly had isolating living situations. A small percentage expressed sadness about their life. In our qualitative analysis, people with DS encouraged parents to love their babies with DS, mentioning that their own lives were good. They further encouraged healthcare professionals to value them, emphasizing that they share similar hopes and dreams as people without DS. Overall, the overwhelming majority of people with DS surveyed indicate they live happy and fulfilling lives.


People's experiences of suffering a lower limb fracture and undergoing surgery.
To describe people's experiences of suffering a lower limb fracture and undergoing surgery, from the time of injury through to the care given at the hospital and recovery following discharge. There is a lack of research on people's experiences of suffering a lower limb fracture and undergoing surgery - from injury to recovery. A qualitative approach was used. Interviews with nine participants were subjected to thematic content analysis. One theme was expressed: from realising the seriousness of the injury to regaining autonomy. Participants described feelings of frustration and helplessness when realising the seriousness of their injury. The wait prior to surgery was a strain and painful experience, and participants needed orientation for the future. They expressed feelings of vulnerability about being in the hands of staff during surgery. After surgery, in the postanaesthesia unit, participants expressed a need to have control and to feel safe in their new situation. To mobilise and regain their autonomy was a struggle, and participants stated that their recovery was extended. Participants found themselves in a new and unexpected situation and experienced pain, vulnerability and a striving for control during the process, that is, 'from realising the seriousness of the injury to regaining autonomy'. How this is managed depends on how the patient's needs are met by nurses. The nursing care received while suffering a lower limb fracture and undergoing surgery should be situation specific as well as individual specific. The safe performance of technical interventions and the nurse's comprehensive explanations of medical terms may help the patient to feel secure during the process.


Focal cerebral hyperemia in acute stroke. Incidence, pathophysiology and clinical significance.
In a consecutive study comprising 41 patients with completed stroke of less than 72 hours duration, cerebral angiography and measurements of the regional cerebral blood flow (rCBF) were performed within 24 hours after admission. The rCBF study was done using the 133-Xenon intracarotid injection method and a 254 multi-detector camera. CT scan was done 24 hours after the rCBF study. Focal cerebral hyperemia was found in 16 patients. The study revealed 3 different types of hyperemia: Border-zone hyperemia, surrounding ischemic areas, was seen in patients with occluded arteries on angiography, presumably resulting from accumulation of acid metabolites in the border-zone of acute infarcts. Postischemic hyperemia was seen in patients without occlusion, presumably due to recanalization of a prior occluded artery. Remote hyperemia was found distant from the infarcted area, presumably due to local tissue pressure on brain tissue. Cortical infarcts (10 patients) all had extensive hyperemic areas. Because the 254 detector camera has an excellent resolution in the cortical surface, our findings strongly suggest that all acute cerebral infarcts are, in fact, associated with hyperemic areas. The hyperemic areas are often extensive and vascular reactivity is commonly impaired. It is suggested that treatment aimed at reducing blood flow in hyperemic areas might improve prognosis.


Histological and immunohistological study of the developing and involuting superficial cervical thymus in the koala (Phascolarctos cinereus).
The thymuses of 44 koalas, ranging from less than 30 d to more than 14 y of age, were examined histologically and immunohistologically. The thymuses from 17 of these koalas dying acutely through trauma were regarded as not being significantly affected by disease and formed the basis for study of the normal thymus. Most other koalas had chronic illness and, consequently, disease affected (involuted) thymuses. Histologically, thymuses showed obvious corticomedullary differentiation with small Hassall's corpuscles visible in koalas more than 8 mo of age. Most cortical and medullary lymphocytes stained for CD3 and CD5 (T lymphocyte markers) while some cells (predominantly medullary) stained for CD79b (B lymphocytes and plasma cells), IgG (plasma cells) or MHC class II (reticular epithelium, macrophages and possibly lymphocytes). Adults of up to 5-6 y of age which had died through trauma had little evidence of involution and had prominent Hassall's corpuscles and medullary epithelial thymocytes. Thymic eosinopoiesis was an inconsistent finding. In traumatised animals over this age, involution was obvious with fibrous replacement of lobules, loss of Hassall's corpuscles and the development of dilated ducts lined by nonciliated epithelium. However, loss of lymphocytes was gradual and pockets of lymphocytes, centrally located in lobules, were still present in the oldest koala examined. In these involuted thymuses, remaining lymphocytes stained for CD3 and lesser numbers of CD5 and CD79b. Plasma cells were common and often stained both for IgG and MHC class II. Thymuses of chronically diseased koalas showed accelerated involution when age matched with thymuses from traumatised koalas. Chronically ill koalas as young as 18-24 mo showed advanced involution, but the morphological and immunohistological characteristics of involuted thymus from diseased koalas could not be distinguished from those of involuted thymuses derived from traumatised koalas. It was concluded that development of the koala thymus is completed at 8 mo of age and that for normal koalas involution is a gradual process which starts not at but after sexual maturity. Immunohistological characterisation of the thymus was comparable to that reported for a variety of eutherian mammals.


Expression of the CYP19 gene and its product aromatase cytochrome P450 in human uterine leiomyoma tissues and cells in culture.
The conversion of C19 steroids to estrogens occurs in a number of tissues and is catalyzed by a specific form of cytochrome P450, namely aromatase cytochrome P450 (P450arom). Previously, conversion of radiolabeled androstenedione to estrone has been demonstrated in uterine leiomyomas. By use of reverse transcription followed by polymerase chain reaction amplification of total RNA together with a rat cRNA as an internal standard, we detected and quantified P450arom transcripts in total RNA isolated from 32 of the 35 leiomyoma (91%) and from 18 of the 24 adjacent myometrial (75%) tissue samples from 26 women. P450arom transcripts were not detectable in myometrial tissues from disease-free uteri (n = 8). P450arom transcript levels in leiomyomas were similar to those in adipose tissue (normalized to total RNA) and were 1.5- to 25-fold higher than those in adjacent myometrial tissues. We did not find any correlation between P450arom transcript levels and leiomyoma size, histopathology, uterine weight, or patient age. In leiomyoma smooth muscle cells in culture (n = 4) and tissue explants (n = 4), aromatase activity was stimulated by dibutyryl cAMP, and this effect was potentiated by a phorbol ester. These increases in aromatase expression were accompanied by comparable increases in the levels of translatable P450arom mRNA. Treatment with dexamethasone or platelet-derived growth factor did not stimulate aromatase expression. Consistently higher levels of aromatase activity and P450arom transcripts were found in the leiomyoma tissues than in smooth muscle cells in culture (2- to 20-fold). Reverse transcription-polymerase chain reaction analysis of untranslated 5'-termini of mRNA species in leiomyomas revealed the use of primarily promoter II (the ovarian-type promoter) for CYP19 gene transcription. Leiomyomas also contain some transcripts with untranslated exon I.4 (previously found in adipose stromal cells and skin fibroblasts). Placental-type promoter-specific 5'-ends were not present in leiomyomas. We conclude that aromatase expression in leiomyomas is regulated by the rate of CYP19 gene transcription, which is, in turn, regulated by the use of tissue-specific promoters. These findings are consistent with the hypothesis that localized estrogen biosynthesis may be of pathological significance in the promotion of leiomyoma growth.


Highly Diverse Aquatic Microbial Communities Separated by Permafrost in Greenland Show Distinct Features According to Environmental Niches.
The Greenland Analog Project (GAP) study area in the vicinity of Kangarlussuaq, Western Greenland, was sampled for surface water and deep groundwater in order to determine the composition and estimate the metabolic features of the microbial communities in water bodies separated by permafrost. The sampling sites comprised a freshwater pond, talik lake, deep anoxic groundwater, glacier ice and supraglacial river, meltwater river and melting permafrost active layer. The microbial communities were characterized by amplicon sequencing of the bacterial and archaeal 16S rRNA genes and fungal ITS1 spacer. In addition, bacterial, archaeal and fungal numbers were determined by qPCR and plate counts, and the utilization pattern of carbon and nitrogen substrates was determined with Biolog AN plates and metabolic functions were predicted with FAPROTAX. Different sample types were clearly distinguishable from each other based on community composition, microbial numbers, and substrate utilization patterns, forming four groups, (1) pond/lake, (2) deep groundwater, (3) glacial ice, and (4) meltwater. Bacteria were the most abundant microbial domain, ranging from 0.2-1.4 × 107 16S rRNA gene copies mL-1 in pond/lake and meltwater, 0.1-7.8 × 106 copies mL-1 in groundwater and less than 104 copies mL-1 in ice. The number of archaeal 16S and fungal 5.8S rRNA genes was generally less than 6.0 × 103 and 1.5 × 103, respectively. N2-fixing and methane-oxidizing Actinomycetes, Bacteroidetes and Verrucomicrobia were the dominant microorganisms in the pond/lake samples, whereas iron reducing Desulfosporosinus sp. dominated the deep anaerobic groundwater. The glacial ice was inhabited by Cyanobacteria, which were mostly Chloroplast-like. The meltwater contained methano- and methylotrophic Proteobacteria, but had also high relative abundances of the nano-sized Parcubacteria. The archaea composed approximately 1% of the 16S rRNA gene pool in the pond/lake samples with nano-sized Woesearchaeota as the dominating taxon, while in the other sample types archaea were almost negligent. Fungi were also most common in the pond/lake communities, were zoospore-forming Chytridiomycetes dominated. Our results show highly diverse microbial communities inhabiting the different cold Greenlandic aqueous environments and show clear segregation of the microbial communities according to habitat, with distinctive dominating metabolic features specifically inhabiting defined environmental niches and a high relative abundance of putatively parasitic or symbiotic nano-sized taxa.


Trends and socioeconomic differences in policy triggers for thinking about quitting smoking: Findings from the International Tobacco Control (ITC) Europe Surveys.
The aim of the current study is to investigate trends and socioeconomic differences in policy triggers for thinking about quitting in six European countries. Data were derived from all available survey waves of the International Tobacco Control (ITC) Europe Surveys (2003-2013). France conducted three survey waves (n=1420-1735), Germany three waves (n=515-1515), The Netherlands seven waves (n=1420-1668), Ireland three waves (n=582-1071), Scotland two waves (n=461-507), and the rest of the United Kingdom conducted seven survey waves (n=861-1737). Smokers were asked whether four different policies (cigarette price, smoking restrictions in public places, free or lower cost medication, and warning labels on cigarette packs) influenced them to think about quitting. Generalized Estimating Equation (GEE) models were estimated for each country. Cigarette price was mentioned most often in all countries and across all waves as trigger for thinking about quitting. Mentioning cigarette price and warning labels increased after the implementation of price increases and warning labels in some countries, while mentioning smoking restrictions decreased after their implementation in four countries. All studied policy triggers were mentioned more often by smokers with low and/or moderate education and income than smokers with high education and income. The education and income differences did not change significantly over time for most policies and in most countries. Tobacco control policies work as a trigger to increase thoughts about quitting, particularly in smokers with low education and low income and therefore have the potential to reduce health inequalities in smoking.


Deep phylogeographic divisions and long-term persistence of forest invertebrates (Hexapoda: Collembola) in the North-Western Mediterranean basin.
The North-Western Mediterranean basin is well known for its high number of relictual endemic taxa, and has been indicated as one of the world's major biodiversity hotspots at the species level. A possible contributing factor may be long-term persistence of populations and their prolonged stability. This study was designed to investigate the phylogeographic structure of three common species of the genus Lepidocyrtus (Hexapoda: Collembola), soil-dwelling arthropods characterized by limited dispersal capabilities and generally associated with forest habitats. We observed a remarkable geographic structure, with numerous deeply divergent genetic lineages occupying islands as well as mainland sites with no apparent gene flow among most sites, even across distances of only tens of kilometres. The reconstructed time frame for the evolution of these lineages suggests divergence between 5 and 15 Ma. This indicates a remarkably ancient origin and long-term persistence of individual lineages over a fine geographic scale despite the occurrence of abrupt sea level and climatic fluctuations in the area. This further suggests that currently recognized morphological species might be a serious underestimation of the true springtail biodiversity within this region.


Internationally comparable diagnosis-specific survival probabilities for calculation of the ICD-10-based Injury Severity Score.
The International Statistical Classification of Diseases, 10th Revision (ICD-10)-based Injury Severity Score (ICISS) performs well but requires diagnosis-specific survival probabilities (DSPs), which are empirically derived, for its calculation. The objective was to examine if DSPs based on data pooled from several countries could increase accuracy, precision, utility, and international comparability of DSPs and ICISS. Australia, Argentina, Austria, Canada, Denmark, New Zealand, and Sweden provided ICD-10-coded injury hospital discharge data, including in-hospital mortality status. Data from the seven countries were pooled using four different methods to create an international collaborative effort ICISS (ICE-ICISS). The ability of the ICISS to predict mortality using the country-specific DSPs and the pooled DSPs was estimated and compared. The pooled DSPs were based on a total of 3,966,550 observations of injury diagnoses from the seven countries. The proportion of injury diagnoses having at least 100 discharges to calculate the DSP varied from 12% to 48% in the country-specific data set and was 66% in the pooled data set. When compared with using a country's own DSPs for ICISS calculation, the pooled DSPs resulted in somewhat reduced discrimination in predicting mortality (difference in c statistic varied from 0.006 to 0.04). Calibration was generally good when the predicted mortality risk was less than 20%. When Danish and Swedish data were used, ICISS was combined with age and sex in a logistic regression model to predict in-hospital mortality. Including age and sex improved both discrimination and calibration substantially, and the differences from using country-specific or pooled DSPs were minor. Pooling data from seven countries generated empirically derived DSPs. These pooled DSPs facilitate international comparisons and enables the use of ICISS in all settings where ICD-10 hospital discharge diagnoses are available. The modest reduction in performance of the ICE-ICISS compared with the country-specific scores is unlikely to outweigh the benefit of internationally comparable Injury Severity Scores possible with pooled data. Prognostic and epidemiological study, level III.


Butyrate modulates diabetes-linked gut dysbiosis: epigenetic and mechanistic modifications.
Diabetic dysbiosis has been described as a novel key player in diabetes and diabetic complications. However, the cellular/molecular alterations associated with dysbiosis remain poorly characterized. For that, control, non-obese type 2 diabetic MKR mice and MKR mice treated with butyrate were used to delineate the epigenetic, cellular and molecular mechanisms by which dysbiosis associated with diabetes induces colon shortening and inflammation attesting to gastrointestinal disturbance. Our results show that dysbiosis is associated with T2DM and characterized by reduced Bacteroid fragilis population and butyrate-forming bacteria. The reduction of butyrate-forming bacteria and inadequate butyrate secretion result in alleviating HDAC3 inhibition and altering colon permeability. The observed changes are also associated with an increase in ROS production, a rise in NOX4 proteins, and a shift in the inflammatory markers, where IL-1β is increased and IL-10 and IL-17α are reduced. Treatment with butyrate restores the homeostatic levels of NOX4 and IL-1β. In summary, our data suggest that in T2DM, dysbiosis is associated with a reduction in butyrate content leading to increased HDAC3 activity. Butyrate treatment restores the homeostatic levels of the inflammatory markers and reduces ROS production known to mediate diabetes-induced colon disturbance. Taken together, our results suggest that butyrate could be a potential treatment to attenuate diabetic complications.


Alternate mechanisms of initial pattern recognition drive differential immune responses to related poxviruses.
Vaccinia immunization was pivotal to successful smallpox eradication. However, the early immune responses that distinguish poxvirus immunization from pathogenic infection remain unknown. To address this, we developed a strategy to map the activation of key signaling networks in vivo and applied this approach to define and compare the earliest signaling events elicited by immunizing (vaccinia) and lethal (ectromelia) poxvirus infections in mice. Vaccinia induced rapid TLR2-dependent responses, leading to IL-6 production, which then initiated STAT3 signaling in dendritic and T cells. In contrast, ectromelia did not induce TLR2 activation, and profound mouse strain-dependent responses were observed. In resistant C57BL/6 mice, the STAT1 and STAT3 pathways were rapidly activated, whereas in susceptible BALB/c mice, IL-6-dependent STAT3 activation did not occur. These data link early immune signaling events to infection outcome and suggest that activation of different pattern-recognition receptors early after infection may be important in determining vaccine efficacy.


State of HIV in the US Deep South.
The Southern United States has been disproportionately affected by HIV diagnoses and mortality. To inform efforts to effectively address HIV in the South, this manuscript synthesizes recent data on HIV epidemiology, care financing, and current research literature on factors that predispose this region to experience a greater impact of HIV. The manuscript focuses on a specific Southern region, the Deep South, which has been particularly affected by HIV. Epidemiologic data from the Centers from Disease Control and Prevention indicate that the Deep South had the highest HIV diagnosis rate and the highest number of individuals diagnosed with HIV (18,087) in 2014. The percentage of new HIV diagnoses that were female has decreased over time (2008-2014) while increasing among minority MSM. The Deep South also had the highest death rates with HIV as an underlying cause of any US region in 2014. Despite higher diagnosis and death rates, the Deep South received less federal government and private foundation funding per person living with HIV than the US overall. Factors that have been identified as contributors to the disproportionate effects of HIV in the Deep South include pervasive HIV-related stigma, poverty, higher levels of sexually transmitted infections, racial inequality and bias, and laws that further HIV-related stigma and fear. Interventions that address and abate the contributors to the spread of HIV disease and the poorer HIV-related outcomes in the Deep South are warranted. Funding inequalities by region must also be examined and addressed to reduce the regional disparities in HIV incidence and mortality.


Biradicals/zwitterions from thermolysis of enyne-isocyanates. Application to the synthesis of 2(1H)-pyridones, benzofuro[3,2-c]pyridin-1(2H)-ones, 2,5-dihydro-1H-pyrido[4,3-b]indol-1-ones, and related compounds.
Thermolysis of benzannulated enyne-isocyanates 13 and enyne-isocyanates 36 and 37 promoted the cycloaromatization reactions to generate in situ O,4-didehydro-2-hydroxyquinolines and O,4-didehydro-2-hydroxypyridines, respectively, as reactive intermediates. These cycloaromatized intermediates could be captured either as biradicals and/or as zwitterions depending on the nature of the substituent at the alkynyl terminus. The intermediate derived from cycloaromatization of 13a bearing a phenyl substituent could be regarded as biradical 14, which then abstracts hydrogen atoms from gamma-terpinene leading to 2(1H)-quinolinone 15. Alternatively, the same intermediate could also be regarded as zwitterion 14', which then undergoes an initial hydride abstraction from gamma-terpinene followed by protonation to produce 15. The presence of a 2-phenylethyl substituent in 13b and 37a or a 2-methylphenyl substituent in 37b also allowed the resulting intermediates to be captured intramolecularly either as biradicals or as zwitterions, producing 2(1H)-quinolinone 19, 2(1H)-pyridone 39, and benzopyranopyridine 43, respectively. On the other hand, with a 2-methoxyphenyl, a 2-(dimethylamino)phenyl, or a 3-methoxypropyl substituent, the chemical behavior of the cycloaromatized adduct could be best accounted for in terms of a zwitterionic intermediate leading to benzofuro[3,2-c]quinolin-6(5H)-one (20), 5,11-dihydro-11-methyl-6H-indolo[3,2-c]quinolin-6-one (25), benzofuro[3,2-c]pyridin-1(2H)-one 44, 2,5-dihydro-2,5-dimethyl-1H-pyrido[4,3-b]indol-1-one 46, and related compounds. Interestingly, thermolysis of 37f bearing a 2-(methoxymethyl)phenyl substituent at the alkynyl terminus produced the unexpected benzopyranopyridine 56 as the major product in a process involving the cleavage of the bond between the methoxyl oxygen and the adjacent methylene carbon. The efficiency and selectivity of the cycloaromatization reaction could also be enhanced by the introduction of 1.1 to 10 equiv of dimethylphenylsilyl chloride to the reaction mixture to capture the resulting zwitterion.


Change in body mass index before and after Alzheimer's disease onset.
A high body mass index (BMI) in middle-age or a decrease in BMI at late-age has been considered a predictor for the development of Alzheimer's disease (AD). However, little is known about the BMI change close to or after AD onset. BMI of participants from three cohorts, the Washington Heights and Inwood Columbia Aging Project (WHICAP; population-based) and the Predictors Study (clinic-based), and National Alzheimer's Coordinating Center (NACC; clinic-based) were analyzed longitudinally. We used generalized estimating equations to test whether there were significant changes of BMI over time, adjusting for age, sex, education, race, and research center. Stratification analyses were run to determine whether BMI changes depended on baseline BMI status. BMI declined over time up to AD clinical onset, with an annual decrease of 0.21 (p=0.02) in WHICAP and 0.18 (p=0.04) kg/m2 in NACC. After clinical onset of AD, there was no significant decrease of BMI. BMI even increased (b=0.11, p=0.004) among prevalent AD participants in NACC. During the prodromal period, BMI decreased over time in overweight (BMI≥25 and <30) WHICAP participants or obese (BMI≥30) NACC participants. After AD onset, BMI tended to increase in underweight/normal weight (BMI<25) patients and decrease in obese patients in all three cohorts, although the results were significant in NACC study only. Our study suggests that while BMI declines before the clinical AD onset, it levels off after clinical AD onset, and might even increase in prevalent AD. The pattern of BMI change may also depend on the initial BMI.


A fusion cytokine coupling GMCSF to IL9 induces heterologous receptor clustering and STAT1 hyperactivation through JAK2 promiscuity.
Cytokine receptors are randomly distributed on the cell surface membrane and are activated upon binding of their extracellular ligands to mediate downstream cellular activities. We hypothesized that pharmaceutical clustering of ligand-bound, activated receptors may lead to heretofore unrealized gain-of-function with therapeutically desirable properties. We here describe an engineered bifunctional cytokine borne of the fusion of Granulocyte Macrophage Colony Stimulating Factor (GMCSF) and Interleukin-9 (IL9) (hereafter GIFT9 fusokine) and demonstrate that it chaperones co-clustering of the functionally unrelated GMCSF receptor (GMCSFR) and IL9 receptor (IL9R) on cell surface of target cells. We demonstrate that GIFT9 treatment of MC/9 cells leads to transhyperphosphorylation of IL9R-associated STAT1 by GMCSFR-associated JAK2. We also show that IL9R-associated JAK1 and JAK3 augment phosphorylation of GMCSFR-linked STAT5. The functional relevance of these synergistic JAK/STAT transphosphorylation events translates to an increased mitogenic response by GMCSFR/IL9R-expressing primary marrow mast cells. The notion of inducing heterologous receptor clustering by engineered fusokines such as GIFT9 opens the door to a novel type of biopharmaceutical platform where designer fusokines modulate cell physiology through clustering of targeted receptor complexes.


Development of doxycycline MIC and disk diffusion interpretive breakpoints and revision of tetracycline breakpoints for Streptococcus pneumoniae.
A study was performed to derive susceptibility testing interpretive breakpoints for doxycycline with Streptococcus pneumoniae and to reassess breakpoints for tetracycline using the requirements defined in Clinical and Laboratory Standards Institute (CLSI) document M23-A3. Tetracycline and doxycycline MICs and disk diffusion zone sizes were determined on 189 isolates selected from the 2009-2010 CDC Active Bacterial Core surveillance strain collection according to the testing methods described in CLSI documents M07-A8 and M02-A10. Tetracycline and doxycycline MICs and zones were compared to each other directly, and the reproducibility of MICs and zone diameters for both drugs was determined. Scattergrams of tetracycline MICs versus corresponding zone diameters and doxycycline MICs versus zones were prepared, and analysis indicated that the present CLSI tetracycline MIC and disk breakpoints did not fit the susceptibility data for doxycycline. Doxycycline was 1 to 3 dilutions more potent than tetracycline, especially in strains harboring the tetM resistance determinant. tetM was detected in ≥ 90% of isolates having tetracycline MICs of ≥ 4 μg/ml and in ≥ 90% with doxycycline MICs of ≥ 1. Limited pharmacokinetic/pharmacodynamic (PK/PD) data coupled with application of the error-rate bounded method of analysis suggested doxycycline-susceptible breakpoints of either ≤ 0.25 μg/ml or ≤ 0.5 μg/ml, with intermediate and resistant breakpoints 1 and 2 dilutions higher, respectively. The disk diffusion zone diameter correlates were susceptible at ≥ 28 mm, intermediate at 25 to 27 mm, and resistant at ≤ 24 mm. Revised lower tetracycline MIC breakpoints were suggested as susceptible at ≤ 1 μg/ml, intermediate at 2 μg/ml, and resistant at ≥ 4 μg/ml. Suggested tetracycline disk diffusion zones were identical to those of doxycycline.


Medical education and the healthcare system--why does the curriculum need to be reformed?
Medical education has been the subject of ongoing debate since the early 1900s. The core of the discussion is about the importance of scientific knowledge on biological understanding at the expense of its social and humanistic characteristics. Unfortunately, reforms to the medical curriculum are still based on a biological vision of the health-illness process. In order to respond to the current needs of society, which is education's main objective, the learning processes of physicians and their instruction must change once again. The priority is the concept of the health-illness process that is primarily social and cultural, into which the biological and psychological aspects are inserted. A new curriculum has been developed that addresses a comprehensive instruction of the biological, psychological, social, and cultural (historical) aspects of medicine, with opportunities for students to acquire leadership, teamwork, and communication skills in order to introduce improvements into the healthcare systems where they work.


Behavioural responses to facial and postural expressions of emotion: An interpersonal circumplex approach.
While the recognition of emotional expressions has been extensively studied, the behavioural response to these expressions has not. In the interpersonal circumplex, behaviour is defined in terms of communion and agency. In this study, we examined behavioural responses to both facial and postural expressions of emotion. We presented 101 Romanian students with facial and postural stimuli involving individuals ('targets') expressing happiness, sadness, anger, or fear. Using an interpersonal grid, participants simultaneously indicated how communal (i.e., quarrelsome or agreeable) and agentic (i.e., dominant or submissive) they would be towards people displaying these expressions. Participants were agreeable-dominant towards targets showing happy facial expressions and primarily quarrelsome towards targets with angry or fearful facial expressions. Responses to targets showing sad facial expressions were neutral on both dimensions of interpersonal behaviour. Postural versus facial expressions of happiness and anger elicited similar behavioural responses. Participants responded in a quarrelsome-submissive way to fearful postural expressions and in an agreeable way to sad postural expressions. Behavioural responses to the various facial expressions were largely comparable to those previously observed in Dutch students. Observed differences may be explained from participants' cultural background. Responses to the postural expressions largely matched responses to the facial expressions.


The co-occurrence of correct and incorrect HIV transmission knowledge and perceived risk for HIV among women of childbearing age in El Salvador.
This article examines the co-occurrence of correct and incorrect knowledge about documented and undocumented modes of HIV transmission among women of childbearing age in El Salvador, and the relationship between HIV transmission knowledge and perceived risk. Incorrect beliefs about HIV transmission co-occur at high levels with, and are largely independent of, accurate knowledge about documented modes of transmission. The co-occurrence of correct and incorrect HIV transmission knowledge was shown to have important implications for perceived risk. Both correct and incorrect HIV transmission knowledge increased the odds of risk perception; uncertainty about risk was decreased among those with higher levels of correct knowledge and increased among those with higher levels of incorrect knowledge. Among those who considered themselves to be at some risk for HIV, higher levels of correct knowledge reduced uncertainty about the degree of risk, while higher levels of incorrect knowledge increased the degree of risk perceived. High levels of endorsement of the documented modes of HIV transmission do not necessarily indicate accurate or adequate knowledge about HIV transmission in the population. Co-occurring inaccurate beliefs about undocumented modes of transmission reflect cultural understandings of contagion and disease, and influence how individuals make sense of medical-scientific information about transmission. Our results suggest that the co-occurrence of correct and incorrect HIV transmission knowledge shapes individual-level risk perceptions. Given the independence of accurate knowledge and inaccurate beliefs. HIV/AIDS education and prevention programs must seek to directly undermine inaccurate beliefs about HIV transmission as part of their efforts to promote behavior change.


Ultra-Early Differential Diagnosis of Acute Cerebral Ischemia and Hemorrhagic Stroke by Measuring the Prehospital Release Rate of GFAP.
Plasma glial fibrillary acidic protein (GFAP) and tau are promising markers for differentiating acute cerebral ischemia (ACI) and hemorrhagic stroke (HS), but their prehospital dynamics and usefulness are unknown. We performed ultra-sensitivite single-molecule array (Simoa®) measurements of plasma GFAP and total tau in a stroke code patient cohort with cardinal stroke symptoms [National Institutes of Health Stroke Scale (NIHSS) ≥3]. Sequential sampling included 2 ultra-early samples, and a follow-up sample on the next morning. We included 272 cases (203 ACI, 60 HS, and 9 stroke mimics). Median (IQR) last-known-well to sampling time was 53 (35-90) minutes for initial prehospital samples, 90 (67-130) minutes for secondary acute samples, and 21 (16-24) hours for next morning samples. Plasma GFAP was significantly higher in patients with HS than ACI (P < 0.001 for <1 hour and <3 hour prehospital samples, and <3 hour secondary samples), while total tau showed no intergroup difference. The prehospital GFAP release rate (pg/mL/minute) occurring between the 2 very early samples was significantly higher in patients with HS than ACI [2.4 (0.6-14.1)] versus 0.3 (-0.3-0.9) pg/mL/minute, P < 0.001. For cases with <3 hour prehospital sampling (ACI n = 178, HS n = 59), a combined rule (prehospital GFAP >410 pg/mL, or prehospital GFAP 90-410 pg/mL together with GFAP release >0.6 pg/mL/minute) enabled ruling out HS with high certainty (NPV 98.4%) in 68% of patients with ACI (sensitivity for HS 96.6%, specificity 68%, PPV 50%). In comparison to single-point measurement, monitoring the prehospital GFAP release rate improves ultra-early differentiation of stroke subtypes. With serial measurement GFAP has potential to improve future prehospital stroke diagnostics.


The effect of work pace on workload, motor variability and fatigue during simulated light assembly work.
This study investigated the effect of work pace on workload, motor variability and fatigue during light assembly work. Upper extremity kinematics and electromyography (EMG) were obtained on a cycle-to-cycle basis for eight participants during two conditions, corresponding to "normal" and "high" work pace according to a predetermined time system for engineering. Indicators of fatigue, pain sensitivity and performance were recorded before, during and after the task. The level and variability of muscle activity did not differ according to work pace, and manifestations of muscle fatigue or changed pain sensitivity were not observed. In the high work pace, however, participants moved more efficiently, they showed more variability in wrist speed and acceleration, but they also made more errors. These results suggest that an increased work pace, within the range addressed here, will not have any substantial adverse effects on acute motor performance and fatigue in light, cyclic assembly work. STATEMENT OF RELEVANCE: In the manufacturing industry, work pace is a key issue in production system design and hence of interest to ergonomists as well as engineers. In this laboratory study, increasing the work pace did not show adverse effects in terms of biomechanical exposures and muscle fatigue, but it did lead to more errors. For the industrial engineer, this observation suggests that an increase in work pace might diminish production quality, even without any noticeable fatigue being experienced by the operators.


Preparing for national school-based deworming in Kenya: the validation and large-scale distribution of school questionnaires with urinary schistosomiasis.
School questionnaires of self-reported schistosomiasis provide a rapid and simple approach for identifying schools at high risk of Schistosoma haematobium and requiring mass treatment. This study investigates the reliability of school questionnaires to identify such schools and infected children within the context of a national school-based deworming programme in Kenya. Between November 2008 and March 2009, 6182 children from 61 schools in Coast Province, Kenya were asked by an interviewer whether they had blood in urine or urinary schistosomiasis (kichocho), and their results were compared with results from microscopic examination of urine samples. Subsequently, in 2009, a school-based questionnaire survey for self-reported schistosomiasis was distributed by the Ministry of Education to all schools in Coast Province, and its results were compared against results from the parasitological survey. The questionnaire survey results were linked to a schools database and mapped. Prevalence of self-reported blood in urine was lower among girls than boys among all ages. The use of a 30% threshold of reported blood in urine was both highly sensitive (91.7%) and specific (100%) in identifying high (>50%) prevalence schools in Coast Province. Questionnaires were however less reliable in diagnosing S. haematobium infection in individuals, particularly among young girls. Comparable levels of reliability were observed when the questionnaire was distributed through the existing education systems and administered by class teachers. The results confirm that blood in urine questionnaires can be reliably used to target mass treatment with praziquantel at national scales. The mapped results of the Ministry of Education survey serve to describe the spatial variation of urinary schistosomiasis and identify schools requiring mass treatment.


Anxiety as a predictor and outcome variable in the multimodal treatment study of children with ADHD (MTA).
Initial moderator analyses in the Multimodal Treatment Study of Children with ADHD (MTA) suggested that child anxiety ascertained by parent report on the Diagnostic Interview Schedule for Children 2.3 (DISC Anxiety) differentially moderated the outcome of treatment. Left unanswered were questions regarding the nature of DISC Anxiety, the impact of comorbid conduct problems on the moderating effect of DISC Anxiety, and the clinical significance of DISC Anxiety as a moderator of treatment outcome. Thirty-three percent of MTA subjects met DSM-III-R criteria for an anxiety disorder excluding simple phobias. Of these, two-thirds also met DSM-III-R criteria for comorbid oppositional-defiant or conduct disorder whereas one-third did not, yielding an odds ratio of approximately two for DISC Anxiety, given conduct problems. In this context, exploratory analyses of baseline data suggest that DISC Anxiety may reflect parental attributions regarding child negative affectivity and associated behavior problems (unlike fearfulness), particularly in the area of social interactions, another core component of anxiety that is more typically associated with phobic symptoms. Analyses using hierarchical linear modeling (HLM) indicate that the moderating effect of DISC Anxiety continues to favor the inclusion of psychosocial treatment for anxious ADHD children irrespective of the presence or absence of comorbid conduct problems. This effect, which is clinically meaningful, is confined primarily to parent-reported outcomes involving disruptive behavior, internalizing symptoms, and inattention; and is generally stronger for combined than unimodal treatment. Contravening earlier studies, no adverse effect of anxiety on medication response for core ADHD or other outcomes in anxious or nonanxious ADHD children was demonstrated. When treating ADHD, it is important to search for comorbid anxiety and negative affectivity and to adjust treatment strategies accordingly.


Structures of the tyrosine kinase domain of fibroblast growth factor receptor in complex with inhibitors.
A new class of protein tyrosine kinase inhibitors was identified that is based on an oxindole core (indolinones). Two compounds from this class inhibited the kinase activity of fibroblast growth factor receptor 1 (FGFR1) and showed differential specificity toward other receptor tyrosine kinases. Crystal structures of the tyrosine kinase domain of FGFR1 in complex with the two compounds were determined. The oxindole occupies the site in which the adenine of adenosine triphosphate binds, whereas the moieties that extend from the oxindole contact residues in the hinge region between the two kinase lobes. The more specific inhibitor of FGFR1 induces a conformational change in the nucleotide-binding loop. This structural information will facilitate the design of new inhibitors for use in the treatment of cancer and other diseases in which cell signaling by tyrosine kinases plays a crucial role in disease pathogenesis.


Effect of growth hormone on the metabolism of thymus and on the immune response against sheep erythrocytes.
The effect of pituitary growth hormone on the biosynthesis of DNA in the thymus and other lymphoid organs, as well as the ability of the rat to respond immunologically to sheep red blood cells, has been evaluated. There is a marked reduction in plaque-forming cells, hemagglutination titers, and DNA synthesis in animals when examined at 15 wk after hypophysectomy. Administration of bovine growth hormone (BGH) leads to the enhancement of DNA synthesis in lymphoid organs and recovery of the immune response. Similar effects of the hormone are observed in plateaued rats. Injection of rabbit anti-BGH globulins, in contrast to normal rabbit globulins, over 5 days causes a drop in the weight of the thymus and in the rate of DNA synthesis in this organ. The thymus is also the organ in which stimulation of DNA synthesis is observed at a time period earlier than the spleen and lymph nodes after a single injection of BGH. The hormone stimulates not only the incorporation of thymidine-(3)H into DNA in the cortical cells, but also the incorporation of sodium sulfate-(35)S into TCA-insoluble biopolymers reported to be elaborated in the medullary area of the thymus. An in vitro system for the action of BGH on the thymus has been described. There is an obligatory requirement for calcium, but not for fetal calf serum in the medium for the hormone effect. An early action of the hormone is the enhanced incorporation of uridine-G-(3)H into RNA in thymocytes which is followed by a stimulation of the synthesis of proteins and DNA. The stimulatory action of growth hormone on RNA synthesis is not because of a facilitated uptake of the radioactive uridine by the cells under hormonal influence, a mechanism by which insulin is observed to increase RNA synthesis in thymocytes in vitro. The action of growth hormone on thymocytes is specific, since thyroid-stimulating hormone (TSH), luteinizing hormone (LH), and heat-inactivated growth hormone are not effective. BGH has also a beneficial action on the regeneration of the thymus and spleen in starved rats.


Creation of a specialist core curriculum for the European Society for Sports traumatology, Knee surgery and Arthroscopy (ESSKA).
The European Society for Sports traumatology, Knee surgery and Arthroscopy (ESSKA) identified the need to develop a core curriculum for clinical specialists that work within the interest areas of ESSKA. A research-based approach was used to define a set of core competencies which could be used to map all of their educational activities, resources and development priorities. This paper describes the aims, development, results and implications of this competency-based core curriculum for orthopaedic conditions relevant to ESSKA members. A Core Curriculum Working Group, with leaders and other experts representing the main specialist areas within ESSKA, reviewed existing curricula and the literature in their own specialist areas. Applying expert group methodology, they iteratively developed a draft list of 285 core competencies for Orthopedic specialists within 6 specialist areas of Knee, Shoulder, Foot/Ankle, Hip, Elbow/Forearm and Sports/Exercise. All ESSKA members were then asked to comment and rate the importance of these competencies, and the Working Group used these findings to critically review and refine the curriculum. The expert groups defined 56 competencies related to 10 Knee pathologies; 67 related to 15 Shoulder pathologies; 45 related to 9 Foot/Ankle pathologies; 41 related to 6 Hip pathologies; and 34 related to 12 Elbow/Forearm pathologies and 42 related to 8 Sports/Exercise pathologies. Survey respondent mean ratings were at least 'Important' for all competencies, and the Working Group used these results to separate the competencies into three levels of importance. A competency-based core curriculum for Orthopedic specialists was achieved through a systematic and scholarly approach, involving both expert opinion and engagement of the wider ESSKA membership, identifying 285 treatment competencies in 6 specialist areas. It is now being used to guide educational and strategic development for ESSKA and should also be of interest to the wider orthopedic and sports medicine communities.


Brodalumab for the Treatment of Moderate-to-Severe Plaque Psoriasis: An Evidence Review Group Evaluation of a NICE Single Technology Appraisal.
As part of the National Institute for Health and Care Excellence single technology appraisal process, brodalumab was assessed to determine the clinical and cost effectiveness of its use in the treatment of moderate-to-severe plaque psoriasis. The Centre for Reviews and Dissemination and the Centre for Health Economics Technology Assessment Group at the University of York were commissioned to act as the independent Evidence Review Group. This article provides a summary of the Evidence Review Group's review of the company's submission, the Evidence Review Group report and the National Institute for Health and Care Excellence Appraisal Committee's subsequent guidance issued in March 2018. The main clinical effectiveness data were derived from three well-conducted, multicentre, double-blind randomised controlled trials. The trials demonstrated that brodalumab statistically significantly reduced the severity of psoriasis and its impact on health-related quality of life, compared with placebo, at 12 weeks. In comparison with ustekinumab, statistically significantly more patients taking brodalumab had reduced psoriasis severity at 12 weeks. Psoriasis severity and quality of life also appeared improved at 52 weeks, although statistical significance was not assessed. Withdrawal rates were comparable to drug survival rates of other biological therapies and rates of adverse events were similar between brodalumab and ustekinumab. A network meta-analysis was presented, comparing brodalumab with other therapies available at the same point in the treatment pathway (i.e. in patients for whom standard systemic therapy or phototherapy is inadequately effective, not tolerated or contraindicated). The network meta-analysis ranked treatments in order of effectiveness, in terms of achieving different levels of Psoriasis Area and Severity Index response. The results indicated that brodalumab had a similar probability of response to ixekizumab, secukinumab and infliximab and a higher probability of response than ustekinumab, adalimumab, etanercept, apremilast, dimethyl fumarate and placebo. The company's economic model compared nine treatment sequences that included three lines of active therapy, consisting of brodalumab and other comparators recommended by the National Institute for Health and Care Excellence, followed by best supportive care. The sequence with brodalumab in the first-line position dominated sequences that started with adalimumab, infliximab, secukinumab and ustekinumab. The incremental cost-effectiveness ratio of the brodalumab sequence compared to less effective and non-dominated sequences ranged from £7145 (vs. the etanercept sequence) to £13,353 (vs. the dimethyl fumarate sequence) per quality-adjusted life-year gained. The incremental cost-effectiveness ratio for the more costly and effective ixekizumab sequence was £894,010 per quality-adjusted life-year gained compared to the brodalumab sequence. At a threshold of £20,000 per quality-adjusted life-year gained, the brodalumab sequence had the highest probability of being cost effective (96%). The main limitation of the company's economic model was the restrictive nature of the sequences compared. Twelve separate scenarios based on key uncertainties were explored by the Evidence Review Group. The only scenarios where brodalumab was ranked lower than first were not considered to be more appropriate or plausible than the assumptions or scenarios included in the company's base case. The treatment rankings identified in the Evidence Review Group's alternative base case were identical to those derived from the company's base case model. At the first National Institute for Health and Care Excellence Appraisal Committee meeting, the Committee concluded that brodalumab appears to be as effective as other anti-interleukin-17 agents and is cost effective, based on the discount agreed in the patient access scheme. Brodalumab is recommended as an option for treating adults with severe plaque psoriasis (defined by a total Psoriasis Area and Severity Index score of 10 or more and a Dermatology Life Quality Index score of more than 10) who have not responded to other systemic non-biological therapies. Brodalumab should be stopped at 12 weeks if the psoriasis has not responded adequately.


Modulation of the hypoxic response following partial bladder outlet obstruction.
Tissue level hypoxia has been noted in animal models of partial bladder outlet obstruction. The key mechanisms linking hypoxia and obstruction induced bladder dysfunction remain unknown. 2-Methoxyestradiol is a natural derivative of 17β-estradiol and is currently used as an oncologic agent for its ability to regulate the hypoxia pathway. We investigated the ability of 2-methoxyestradiol to modulate the hypoxia response in a mouse model of bladder obstruction. A group of 5 to 6-week-old female C57BL/6 mice underwent oophorectomy and partial bladder outlet obstruction. Obstructed animals received a subcutaneous pellet of cholesterol placebo (7) or 2-methoxyestradiol plus cholesterol (7). Age matched controls underwent oophorectomy only (8). After 4 weeks the bladders of mice with partial bladder outlet obstruction and of unobstructed animals were harvested. Bladder sections (5 μm) were immunostained for Hypoxyprobe™-1, glucose transporter 1 and hypoxia inducible factor-1α. Real-time polymerase chain reaction was performed for hypoxia inducible factor-1α and lysyl oxidase. Statistical analysis was performed using 1-way ANOVA and the Wilcoxon rank sum test. Immunostaining for glucose transporter 1 and Hypoxyprobe-1 revealed the presence of tissue hypoxia after partial bladder outlet obstruction. Immunostaining and real-time polymerase chain reaction demonstrated the up-regulation of hypoxia inducible factor-1α in mice after partial bladder outlet obstruction compared to controls (p = 0.0394). Although not statistically significant, a trend toward lower gene expression of hypoxia inducible factor-1α was seen in mice receiving 2-methoxyestradiol compared to placebo (p = 0.0625). Compared to placebo, 2-methoxyestradiol treatment increased lysyl oxidase expression (p = 0.007). Murine partial bladder outlet obstruction resulted in hypoxia and up-regulation of the hypoxia inducible factor-1 pathway. Subcutaneous 2-methoxyestradiol administration attenuated this response and may be a viable tool to study the role of hypoxia after partial bladder outlet obstruction.


Comparative genomic analysis of prion genes.
The homologues of human disease genes are expected to contribute to better understanding of physiological and pathogenic processes. We made use of the present availability of vertebrate genomic sequences, and we have conducted the most comprehensive comparative genomic analysis of the prion protein gene PRNP and its homologues, shadow of prion protein gene SPRN and doppel gene PRND, and prion testis-specific gene PRNT so far. While the SPRN and PRNP homologues are present in all vertebrates, PRND is known in tetrapods, and PRNT is present in primates. PRNT could be viewed as a TE-associated gene. Using human as the base sequence for genomic sequence comparisons (VISTA), we annotated numerous potential cis-elements. The conserved regions in SPRNs harbour the potential Sp1 sites in promoters (mammals, birds), C-rich intron splicing enhancers and PTB intron splicing silencers in introns (mammals, birds), and hsa-miR-34a sites in 3'-UTRs (eutherians). We showed the conserved PRNP upstream regions, which may be potential enhancers or silencers (primates, dog). In the PRNP 3'-UTRs, there are conserved cytoplasmic polyadenylation element sites (mammals, birds). The PRND core promoters include highly conserved CCAAT, CArG and TATA boxes (mammals). We deduced 42 new protein primary structures, and performed the first phylogenetic analysis of all vertebrate prion genes. Using the protein alignment which included 122 sequences, we constructed the neighbour-joining tree which showed four major clusters, including shadoos, shadoo2s and prion protein-likes (cluster 1), fish prion proteins (cluster 2), tetrapode prion proteins (cluster 3) and doppels (cluster 4). We showed that the entire prion protein conformationally plastic region is well conserved between eutherian prion proteins and shadoos (18-25% identity and 28-34% similarity), and there could be a potential structural compatibility between shadoos and the left-handed parallel beta-helical fold. It is likely that the conserved genomic elements identified in this analysis represent bona fide cis-elements. However, this idea needs to be confirmed by functional assays in transgenic systems.


Pharmacogenetics of treatments for pancreatic cancer.
Introduction: Despite clinical efforts, pancreatic ductal adenocarcinoma (PDAC) has a dismal prognosis. The scarcity of effective therapies can be reflected by the lack of reliable biomarkers to adapt anticancer drugs prescription to tumors' and patients' features. Areas covered: Pharmacogenetics should provide the way to select patients who may benefit from a specific therapy that best matches individual and tumor genetic profile, but it has not yet led to gains in outcome. This review describes PDAC pharmacogenetics findings, critically reappraising studies on polymorphisms and -omics profiles correlated to response to gemcitabine, FOLFIRINOX, and nab-paclitaxel combinations, as well as limitations of targeted therapies. Further, we question whether personalized approaches will benefit patients to any significant degree, supporting the need of new strategies within well-designed trials and validated genomic tests for treatment decision-making. Expert opinion: A major challenge in PDAC is the identification of subgroups of patients who will benefit from treatments. Minimally-invasive tests to analyze biomarkers of drug sensitivity/toxicity should be developed alongside anticancer treatments. However, progress might fall below expectations because of tumor heterogeneity and clonal evolution. Whole-genome sequencing and liquid biopsies, as well as prospective validation in selected cohorts, should overcome the limitations of traditional pharmacogenetic approaches.


Transmission of Rift Valley fever virus from European-breed lambs to Culex pipiens mosquitoes.
Rift Valley fever virus (RVFV) is a mosquito-borne bunyavirus of the genus Phlebovirus that is highly pathogenic to ruminants and humans. The disease is currently confined to Africa and the Arabian Peninsula, but globalization and climate change may facilitate introductions of the virus into currently unaffected areas via infected animals or mosquitoes. The consequences of such an introduction will depend on environmental factors, the availability of susceptible ruminants and the capacity of local mosquitoes to transmit the virus. We have previously demonstrated that lambs native to the Netherlands are highly susceptible to RVFV and we here report the vector competence of Culex (Cx.) pipiens, the most abundant and widespread mosquito species in the country. Vector competence was first determined after artificial blood feeding of laboratory-reared mosquitoes using the attenuated Clone 13 strain. Subsequently, experiments with wild-type RVFV and mosquitoes hatched from field-collected eggs were performed. Finally, the transmission of RVFV from viremic lambs to mosquitoes was studied. Artificial feeding experiments using Clone 13 demonstrated that indigenous, laboratory-reared Cx. pipiens mosquitoes are susceptible to RVFV and that the virus can be transmitted via their saliva. Experiments with wild-type RVFV and mosquitoes hatched from field-collected eggs confirmed the vector competence of Cx. pipiens mosquitoes from the Netherlands. To subsequently investigate transmission of the virus under more natural conditions, mosquitoes were allowed to feed on RVFV-infected lambs during the viremic period. We found that RVFV is efficiently transmitted from lambs to mosquitoes, although transmission was restricted to peak viremia. Interestingly, in the mosquito-exposed skin samples, replication of RVFV was detected in previously unrecognized target cells. We here report the vector competence of Cx. pipiens mosquitoes from the Netherlands for RVFV. Both laboratory-reared mosquitoes and well as those hatched from field-collected eggs were found to be competent vectors. Moreover, RVFV was transmitted efficiently from indigenous lambs to mosquitoes, although the duration of host infectivity was found to be shorter than previously assumed. Interestingly, analysis of mosquito-exposed skin samples revealed previously unidentified target cells of the virus. Our findings underscore the value of including natural target species in vector competence experiments.