Systematic literature review of oral hygiene practices for intensive care patients receiving mechanical ventilation.
Oropharyngeal colonization with pathogenic organisms contributes to the development of ventilator-associated pneumonia in intensive care units. Although considered basic and potentially nonessential nursing care, oral hygiene has been proposed as a key intervention for reducing ventilator-associated pneumonia. Nevertheless, evidence from randomized controlled trials that could inform best practice is limited. To appraise the peer-reviewed literature to determine the best available evidence for providing oral care to intensive care patients receiving mechanical ventilation and to document a research agenda for this important activity in optimizing patients' outcomes. Articles published from 1985 to 2006 in English and indexed in the CINAHL, MEDLINE, Joanna Briggs Institute, Cochrane Library, EMBASE, and DARE databases were searched by using the key terms oral hygiene, oral hygiene practices, oral care, mouth care, mouth hygiene, intubated, mechanically ventilated, intensive care, and critical care. Reference lists of retrieved journal articles were searched for publications missed during the primary search. Finally, the Google search engine was used to do a comprehensive search of the World Wide Web to ensure completeness of the search. The search strategy was verified by a health librarian. The search yielded 55 articles: 11 prospective controlled trials, 20 observational studies, and 24 descriptive reports. Methodological issues and the heterogeneity of samples precluded meta-analysis. Despite the importance of providing oral hygiene to intensive care patients receiving mechanical ventilation, high-level evidence from rigorous randomized controlled trials or high-quality systematic reviews that could inform clinical practice is scarce.


Nuclear actin regulates dynamic subcellular localization and activity of the SRF cofactor MAL.
Actin, which is best known as a cytoskeletal component, also participates in the control of gene expression. We report a function of nuclear actin in the regulation of MAL, a coactivator of the transcription factor serum response factor (SRF). MAL, which binds monomeric actin, is cytoplasmic in many cells but accumulates in the nucleus upon serum-induced actin polymerization. MAL rapidly shuttles between cytoplasm and nucleus in unstimulated cells. Serum stimulation effectively blocks MAL nuclear export, which requires MAL-actin interaction. Nuclear MAL binds SRF target genes but remains inactive unless actin binding is disrupted. Fluorescence resonance energy transfer analysis demonstrates that the MAL-actin interaction responds to extracellular signals. Serum-induced signaling is thus communicated to nuclear actin to control a transcriptional regulator.


Peer Supports for Tobacco Cessation for Adults with Serious Mental Illness: A Review of the Literature.
OBJECTIVE: A relatively new approach to addressing tobacco use in mental health settings is the involvement of consumers or peers, defined as other individuals with mental health conditions, as service providers. This review examines the literature describing peer delivered supports for tobacco cessation for adults with serious mental illness. METHODS: The authors conducted a review of online databases to identify reports of tobacco cessation interventions in which peers play a key role in the provision of services to help other adults with serious mental illness to reduce or quit using tobacco. RESULTS: Our review yielded reports of four tobacco cessation interventions for persons with serious mental illness that include peer providers. The roles of peers in these interventions include that of co-leaders of an educational smoking cessation group, individual counselors as part of a multi-faceted tobacco treatment program, and as outreach tobacco cessation advocates. CONCLUSIONS: The roles of peers in these interventions are promising; however, more detail about the selection and experience of the peer providers, as well as the services they deliver in tobacco cessation interventions, would be helpful. In addition, rigorous research is needed to evaluate the impact of peer providers on reducing tobacco use in this population.


Childhood onset of stiff-man syndrome.
Reports of pediatric-onset stiff-man syndrome (SMS) are rare. This may be an underrecognized disorder in child neurology practice. To describe patients with disorders in the SMS spectrum beginning in childhood. This study was a medical record review and serological evaluation conducted at child and adult neurology clinics at the Mayo Clinic, Rochester, Minnesota. Systematic review of the literature was conducted of patients who presented from 1984-2012 with onset of symptomatic SMS occurring at age 18 years or younger. Response to symptomatic and immunotherapies, patient and physician reported, including modified Rankin scale. We identified 8 patients with childhood-onset SMS, representing 5% of patients with SMS evaluated at Mayo Clinic during a period of 29 years (4 were girls). The median age at symptom onset was 11 years (range, 1-14 years). The diagnosis in 3 patients was not established until adulthood (median symptom duration at diagnosis, 14 years; range, 0-46 years). The phenotypes encountered were: classic SMS (n = 5, involving the low back and lower extremities), variant SMS (n = 2, limited to 1 limb [with dystonic posture] or back), and progressive encephalomyelitis with rigidity and myoclonus (n = 1). Initial misdiagnoses included functional movement disorder (n = 2), generalized dystonia and parkinsonism (n = 1), and hereditary spastic paraparesis (n = 1). Six patients had 1 or more coexisting autoimmune disorders: type 1 diabetes mellitus (n = 4), thyroid disease (n = 2), and vitiligo (n = 2). Serologic study results revealed glutamic acid decarboxylase 65-IgG in all cases (median value, 754 nmol/L; range, 0.06-3847 nmol/L; normal value, ≤ 0.02 nmol/L) and glycine receptor antibody in 3 cases. Improvements were noted with symptomatic therapy (diazepam, 6 of 6 patients treated, and oral baclofen, 3 of 3 treated) and immunotherapy (intravenous immune globulin, 3 of 4 treated and plasmapheresis, 3 of 4 treated). The 3 patients with glycine receptor antibody all improved with immunotherapy. At last follow-up, 4 patients had mild or no symptoms, but 4 had moderate or severe residual symptoms and required maintenance symptomatic therapy (n = 5) and immunotherapy (n = 4). Ten of 12 pediatric SMS cases identified by literature review had a severe whole-body phenotype resembling progressive encephalomyelitis with rigidity and myoclonus. Childhood-onset SMS is a rare but underrecognized and treatable disorder. Serological and electrophysiological testing aid diagnosis.


Factors influencing movement probabilities of big brown bats (Eptesicus fuscus) in buildings.
We investigated movements of female big brown bats (Eptesicus fuscus) roosting in maternity colonies in buildings in Fort Collins, Colorado (USA), during the summers of 2002, 2003, and 2005. This behavior can be of public health concern where bats that may carry diseases (e.g., rabies) move among buildings occupied by people. We used passive integrated transponders (PIT tags) to mark individual bats and hoop PIT readers at emergence points to passively monitor the use of building roosts by marked adult females on a daily basis during the lactation phase of reproduction. Multi-strata models were used to examine movements among roosts in relation to ambient temperatures and ectoparasite loads. Our results suggest that high ambient temperatures influence movements. Numbers of mites (Steatonyssus occidentalis) did not appear to influence movements of female bats among building roosts. In an urban landscape, periods with unusually hot conditions are accompanied by shifting of bats to different buildings or segments of buildings, and this behavior may increase the potential for contact with people in settings where, in comparison to their more regularly used buildings, the bats may be more likely to be of public concern as nuisances or health risks.


Menopausal Hormone Therapy Influence on Breast Cancer Outcomes in the Women's Health Initiative.
The Woman's Health Initiative has conducted 2 full-scale, placebo-controlled clinical trials to determine the influence of menopausal therapy on breast cancer incidence and outcome. Estrogen plus progestin use in postmenopausal women with a uterus increases breast cancer incidence and deaths from breast cancer. Despite a short-term reduction in risk after stopping estrogen plus progestin use, an increase in breast cancer risk persists postintervention. Estrogen-alone use in postmenopausal women with prior hysterectomy reduces breast cancer incidence and reduces deaths from breast cancer. The reduced breast cancer risk persists for several years after stopping estrogen-alone use but is lost in late postintervention. These findings suggest recalibration of breast cancer risk and benefit consideration for both regimens, with estrogen plus progestin use associated with greater risk and estrogen-alone use associated with greater benefit. Use of either regimen in clinical practice requires careful consideration of all clinical risks and benefits.


Genes with 5' terminal oligopyrimidine tracts preferentially escape global suppression of translation by the SARS-CoV-2 Nsp1 protein.
Viruses rely on the host translation machinery to synthesize their own proteins. Consequently, they have evolved varied mechanisms to co-opt host translation for their survival. SARS-CoV-2 relies on a nonstructural protein, Nsp1, for shutting down host translation. However, it is currently unknown how viral proteins and host factors critical for viral replication can escape a global shutdown of host translation. Here, using a novel FACS-based assay called MeTAFlow, we report a dose-dependent reduction in both nascent protein synthesis and mRNA abundance in cells expressing Nsp1. We perform RNA-seq and matched ribosome profiling experiments to identify gene-specific changes both at the mRNA expression and translation levels. We discover that a functionally coherent subset of human genes is preferentially translated in the context of Nsp1 expression. These genes include the translation machinery components, RNA binding proteins, and others important for viral pathogenicity. Importantly, we uncovered a remarkable enrichment of 5' terminal oligo-pyrimidine (TOP) tracts among preferentially translated genes. Using reporter assays, we validated that 5' UTRs from TOP transcripts can drive preferential expression in the presence of Nsp1. Finally, we found that LARP1, a key effector protein in the mTOR pathway, may contribute to preferential translation of TOP transcripts in response to Nsp1 expression. Collectively, our study suggests fine-tuning of host gene expression and translation by Nsp1 despite its global repressive effect on host protein synthesis.


Feedback-mediated dynamics in a model of a compliant thick ascending limb.
The tubuloglomerular feedback (TGF) system in the kidney, which is a key regulator of filtration rate, has been shown in physiologic experiments in rats to mediate oscillations in tubular fluid pressure and flow, and in NaCl concentration in the tubular fluid of the thick ascending limb (TAL). In this study, we developed a mathematical model of the TGF system that represents NaCl transport along a TAL with compliant walls. The model was used to investigate the dynamic behaviors of the TGF system. A bifurcation analysis of the TGF model equations was performed by deriving and finding roots of the characteristic equation, which arises from a linearization of the model equations. Numerical simulations of the full model equations were conducted to assist in the interpretation of the bifurcation analysis. These techniques revealed a complex parameter region that allows a variety of qualitatively different model solutions: a regime having one stable, time-independent steady-state solution; regimes having one stable oscillatory solution only; and regimes having multiple possible stable oscillatory solutions. Model results suggest that the compliance of the TAL walls increases the tendency of the model TGF system to oscillate.


Bioassay of Reserpine for Possible Carcinogenicity (CAS No. 50-55-5).
A bioassay for possible carcinogenicity of reserpine, an antihypertensive drug for human use, was conducted by administering the test chemical in feed to F344 rats and B6C3F1 mice. Groups of 50 rats and 50 mice of each sex were administered reserpine at two doses, 5 ppm and 10 ppm, for 103 weeks and then observed for an additional 2 weeks. Matched controls consisted of groups of 50 untreated rats and 50 untreated mice of each sex. All surviving animals were killed and necropsied at the end of 104 or 105 weeks. The significant effects that could be related to administration of reserpine at the doses used were decreased body weight and increased tumor formation in dosed male rats and in mice of both sexes. Dosed male rats had an increased incidence of adrenal medullary pheochromocytomas. Among dosed mice, some males developed undifferentiatedcarcinomas of the seminal vesicles, which rarely occur in control mice, and females had an increased incidence of mammary cancer. It was concluded that, under the conditions of the bioassay, reserpine was carcinogenic in male rats and in mice of both sexes, producing three different kinds of cancers. reserpine was not carcinogenic for female rats, but they may not have received a high enough dose for maximum test sensitivity. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Negative Male Mice: Positive Female Mice: Positive


The polycystic ovary syndrome: pathogenesis and treatment.
To propose a theory for the pathogenesis of the polycystic ovary syndrome that explains the endocrinologic abnormalities of the syndrome and provides a rational approach to therapy. An English-language literature search using MEDLINE (1975 to 1988) and extensive bibliographic reviews of identified articles. We reviewed the literature and selected 169 articles considered most relevant for the definition of the endocrinologic abnormalities, elucidation of pathogenic mechanisms, or delineation of therapeutic interventions. The authors independently assessed study quality and data concerning endocrinologic abnormalities, pathogenic mechanisms, and therapy of the polycystic ovary syndrome. The polycystic ovary syndrome may be best defined as functional, gonadotropin-dependent ovarian hyperandrogenism. The polycystic ovary syndrome results when a primary defect increases one of three variables: the ratio of the serum concentrations of luteinizing hormone to follicle stimulating hormone, the ratio of the intraovarian concentrations of androgen to estrogen, or follicular atresia. An increase in one of these variables can then induce successive abnormalities in one or more of the remaining two variables in a to-and-fro manner. Evidence suggests that several causes exist, each of which can produce the clinical syndrome by a to-and-fro interaction among pathogenic mechanisms. Treatment objectives for the syndrome include amelioration of hirsutism, induction of ovulation, and weight reduction.


Selective coupling of HE11 and TM01 modes into microfabricated fully metal-coated quartz probes.
We report computational and experimental investigations on injection and transmission of light in microfabricated fully Aluminum-coated quartz probes. In particular, we show that a selective coupling of either the HE(11) or the TM(01) mode can be carried out by injecting focused linearly or radially polarized beams into the probe. Optical fields, emitted by the probe after a controlled injection, are characterized in intensity and phase with the help of an interferometric technique. With the help of near-field measurement, we finally demonstrate that a longitudinally polarized spot localized at the tip apex is actually produced when the TM(01) mode is coupled into the probe.


Cerebellar hemorrhage as a first presentation of acquired Hemophilia A.
Acquired hemophilia A (AHA) is an uncommon coagulation disorder caused by the development of autoantibodies against coagulation factor VIII (FVIII). While intracranial hemorrhage is a known complication of AHA, intracranial hemorrhage as the presenting manifestation of AHA has only been described in three previous case reports. We report a case of an 86-year-old woman with no previously reported history of coagulopathy presenting with an acute intraparenchymal cerebellar hemorrhage and laboratory studies demonstrating an isolated prolonged activated partial thromboplastin time (aPTT). We discuss an approach to the prolonged aPTT, and review the literature concerning the diagnosis and treatment of AHA. Occipital decompressive craniectomy with evacuation of the hemorrhage was performed. Eight hours following the procedure, the patient's status acutely declined with demonstration of a reoccurrence of the cerebellar hemorrhage and new right frontal lobe hemorrhage. After discussion with the patient's family, life-sustaining support measures were withdrawn. Postmortem analysis revealed a low FVIII activity level and the presence of FVIII inhibitor. The presentation of intracranial hemorrhage with an isolated prolonged aPTT is concerning for an acquired hemophilia with FVIII deficiency. Other causes of isolated prolonged aPTT such as a lupus anticoagulant must also be considered. Preoperative identification and work-up of the coagulation abnormality is essential to guide initial treatment.


The COVID-19 pandemic altered the modality, but not the frequency, of formal cognitive assessment.
We investigated the impacts of the COVID-19 pandemic on the modality of formal cognitive assessments (in-person versus remote assessments). We created a web-based survey with 34 items and collected data from 114 respondents from a range of health care professions and settings. We established the proportion of cognitive assessments which were face-to-face or via video or telephone conferencing, both pre- and post-March 2020. Further, we asked respondents about the assessment tools used and perceived barriers, challenges, and facilitators for the remote assessment of cognition. In addition, we asked questions specifically about the use of the Oxford Cognitive Screen. We found that the frequency of assessing cognition was stable compared to pre-pandemic levels. Use of telephone and video conferencing cognitive assessments increased by 10% and 18% respectively. Remote assessment increased accessibility to participants and safety but made observing the subtleties of behaviour during test administration difficult. The respondents called for an increase in the availability of standardised, validated, and normed assessments. We conclude that the pandemic has not been detrimental to the frequency of cognitive assessments. In addition, a shift in clinical practice to include remote cognitive assessments is clear and wider availability of validated and standardised remote assessments is necessary.IMPLICATIONS FOR REHABILITATIONWe caution the wider use and interpretation of remote formal cognitive assessments due to lack of validated, standardised, and normed assessments in a remote format.Clinicians should seek out the latest validation and normative data papers to ensure they are using the most up to date tests and respective cut offs.Support is needed for individuals who lack knowledge/have anxiety over the use of technology in formal cognitive assessments.


High-Resolution Intracranial Pressure Burden and Outcome in Subarachnoid Hemorrhage.
Intracranial pressure (ICP) control is a therapeutic target in patients with aneurysmal subarachnoid hemorrhage, although only a limited number of studies assessed its course and effect on outcome. Pressure-time dose (PTDICP) is a method to quantify the burden and the time spent above a defined threshold of ICP. PTDICP or its relationship with outcome has never been evaluated in aneurysmal subarachnoid hemorrhage. Analysis of data prospectively collected from aneurysmal subarachnoid hemorrhage patients admitted to Neurointensive Care Unit. Monitored data, including intraparenchymal ICP, were digitally recorded minute-by-minute in the first 7 days. PTDICP (mm Hg h) was computed using 4 predefined thresholds (15, 20, 25, and 30 mm Hg). Outcome was assessed through Extended Glasgow Outcome Scale at hospital discharge and at 6 months. Fifty-five patients were enrolled. Forty-two patients (76%) presented with a poor clinical grade. Overall, mortality was 17% at hospital discharge and 34% at 6 months. Half of patients required extensive therapy to control high ICP during day 1. Median ICP was 10 mm Hg (4-75), whereas median PTDICP15, PTDICP20, PTDICP25, PTDICP30 were, respectively, 13, 4, 2, and 1 mm Hg h. We observed an association between mortality at hospital discharge and higher level of PTDICP using 20, 25, and 30 mm Hg as thresholds and between exposure to a moderate-level PTDICP30 and unfavorable long-term outcome. PTDICP may better define one of the insults that the brain suffers after aneurysmal rupture, and exposure to moderate PTDICP30 was significant prognostic factor of 6-month unfavorable outcome.


Conflict-free collective stochastic decision making by orbital angular momentum of photons through quantum interference.
In recent cross-disciplinary studies involving both optics and computing, single-photon-based decision-making has been demonstrated by utilizing the wave-particle duality of light to solve multi-armed bandit problems. Furthermore, entangled-photon-based decision-making has managed to solve a competitive multi-armed bandit problem in such a way that conflicts of decisions among players are avoided while ensuring equality. However, as these studies are based on the polarization of light, the number of available choices is limited to two, corresponding to two orthogonal polarization states. Here we propose a scalable principle to solve competitive decision-making situations by using the orbital angular momentum of photons based on its high dimensionality, which theoretically allows an unlimited number of arms. Moreover, by extending the Hong-Ou-Mandel effect to more than two states, we theoretically establish an experimental configuration able to generate multi-photon states with orbital angular momentum and conditions that provide conflict-free selections at every turn. We numerically examine total rewards regarding three-armed bandit problems, for which the proposed strategy accomplishes almost the theoretical maximum, which is greater than a conventional mixed strategy intending to realize Nash equilibrium. This is thanks to the quantum interference effect that achieves no-conflict selections, even in the exploring phase to find the best arms.


Accuracy of the general practitioner's sense of alarm when confronted with dyspnoea and/or chest pain: a prospective observational study.
Dyspnoea and chest pain are symptoms shared with multiple pathologies ranging from the benign to life-threatening diseases. A Gut Feelings Questionnaire (GFQ) has been validated to measure the general practitioner's (GPs) sense of alarm or sense of reassurance. The aim of the study was to estimate the diagnostic test accuracy of GPs' sense of alarm when confronted with dyspnoea and chest pain. Prospective observational study in general practice. Patients aged between 18 and 80 years, consulting their GP for dyspnoea and/or chest pain, were considered for enrolment. These GPs had to complete the GFQ immediately after the consultation. Life-threatening and non-life-threatening diseases have previously been defined according to the pathologies or symptoms in the International Classification of Primary Care (ICPC)-2 classification. The index test was the sense of alarm and the reference standard was the final diagnosis at 4 weeks. 25 GPs filled in 235 GFQ questionnaires. The positive likelihood ratio for the sense of alarm was 2.12 (95% CI 1.49 to 2.82), the negative likelihood ratio was 0.55 (95% CI 0.37 to 0.77). Where the physician experienced a sense of alarm when a patient consulted him/her for dyspnoea and/or chest pain, the post-test odds that this patient had, in fact, a life-threatening disease was about twice as high as the pretest odds. NCT02932982.


The Balance protocol: a pragmatic weight gain prevention randomized controlled trial for medically vulnerable patients within primary care.
For patients with obesity who are not ready for or experience barriers to weight loss, clinical practice guidelines recommend provider counseling on preventing further weight gain as a first-line treatment approach. Unfortunately, evidence-based weight gain prevention interventions are not routinely available within primary care. To address this gap, we will implement a pragmatic 12-month randomized controlled trial of a digital weight gain prevention intervention delivered to patients receiving primary care within a network of Federally Qualified Community Health Centers in central North Carolina. Balance (Equilibrio in Spanish) is a pragmatic effectiveness trial that will randomize adult patients who have overweight or obesity (BMI of 25-40 kg/m2) to either: 1) a weight gain prevention intervention with tailored behavior change goals and tracking, daily weighing on a network-connected electronic scale, and responsive weight and goal coaching delivered remotely by health center registered dietitians; or 2) a usual care program with automated healthy living text messages and print materials and routine primary care. The primary outcome will be weight gain prevention at 24-months, defined as ≤3% change in baseline weight. To align with its pragmatic design, trial outcome data will be pulled from the electronic health record of the community health center network. For underserved, often rurally-located patients with obesity, digital approaches to promote a healthy lifestyle can curb further weight gain. Yet enrolling medically vulnerable patients into a weight gain prevention trial, many of whom are from racial/ethnic minorities, can be difficult. Despite these potential challenges, we plan to recruit a large, diverse sample from rural areas, and will implement a remotely-delivered weight gain prevention intervention to medically vulnerable patients. Upcoming trial results will demonstrate the effectiveness of this pragmatic approach to implement and evaluate a digital weight gain prevention intervention within primary care. NCT03003403 . Registered December 28, 2016.


Electrostatics of aquaporin and aquaglyceroporin channels correlates with their transport selectivity.
Aquaporins are homotetrameric channel proteins, which allow the diffusion of water and small solutes across biological membranes. According to their transport function, aquaporins can be divided into "orthodox aquaporins", which allow the flux of water molecules only, and "aquaglyceroporins", which facilitate the diffusion of glycerol and other small solutes in addition to water. The contribution of individual residues in the pore to the selectivity of orthodox aquaporins and aquaglyceroporins is not yet fully understood. To gain insights into aquaporin selectivity, we focused on the sequence variation and electrostatics of their channels. The continuum Poisson-Boltzmann electrostatic potential along the channel was calculated and compared for ten three-dimensional-structures which are representatives of different aquaporin subfamilies, and a panel of functionally characterized mutants, for which high-accuracy three-dimensional-models could be derived. Interestingly, specific electrostatic profiles associated with the main selectivity to water or glycerol could be identified. In particular: (i) orthodox aquaporins showed a distinctive electrostatic potential maximum at the periplasmic side of the channel around the aromatic/Arg (ar/R) constriction site; (ii) aquaporin-0 (AQP0), a mammalian aquaporin with considerably low water permeability, had an additional deep minimum at the cytoplasmic side; (iii) aquaglyceroporins showed a rather flat potential all along the channel; and (iv) the bifunctional protozoan PfAQP had an unusual all negative profile. Evaluation of electrostatics of the mutants, along with a thorough sequence analysis of the aquaporin pore-lining residues, illuminated the contribution of specific residues to the electrostatics of the channels and possibly to their selectivity.


Health benefits, costs, and cost-effectiveness of earlier eligibility for adult antiretroviral therapy and expanded treatment coverage: a combined analysis of 12 mathematical models.
New WHO guidelines recommend initiation of antiretroviral therapy for HIV-positive adults with CD4 counts of 500 cells per μL or less, a higher threshold than was previously recommended. Country decision makers have to decide whether to further expand eligibility for antiretroviral therapy accordingly. We aimed to assess the potential health benefits, costs, and cost-effectiveness of various eligibility criteria for adult antiretroviral therapy and expanded treatment coverage. We used several independent mathematical models in four settings-South Africa (generalised epidemic, moderate antiretroviral therapy coverage), Zambia (generalised epidemic, high antiretroviral therapy coverage), India (concentrated epidemic, moderate antiretroviral therapy coverage), and Vietnam (concentrated epidemic, low antiretroviral therapy coverage)-to assess the potential health benefits, costs, and cost-effectiveness of various eligibility criteria for adult antiretroviral therapy under scenarios of existing and expanded treatment coverage, with results projected over 20 years. Analyses assessed the extension of eligibility to include individuals with CD4 counts of 500 cells per μL or less, or all HIV-positive adults, compared with the previous (2010) recommendation of initiation with CD4 counts of 350 cells per μL or less. We assessed costs from a health-system perspective, and calculated the incremental cost (in US$) per disability-adjusted life-year (DALY) averted to compare competing strategies. Strategies were regarded very cost effective if the cost per DALY averted was less than the country's 2012 per-head gross domestic product (GDP; South Africa: $8040; Zambia: $1425; India: $1489; Vietnam: $1407) and cost effective if the cost per DALY averted was less than three times the per-head GDP. In South Africa, the cost per DALY averted of extending eligibility for antiretroviral therapy to adult patients with CD4 counts of 500 cells per μL or less ranged from $237 to $1691 per DALY averted compared with 2010 guidelines. In Zambia, expansion of eligibility to adults with a CD4 count threshold of 500 cells per μL ranged from improving health outcomes while reducing costs (ie, dominating the previous guidelines) to $749 per DALY averted. In both countries results were similar for expansion of eligibility to all HIV-positive adults, and when substantially expanded treatment coverage was assumed. Expansion of treatment coverage in the general population was also cost effective. In India, the cost for extending eligibility to all HIV-positive adults ranged from $131 to $241 per DALY averted, and in Vietnam extending eligibility to patients with CD4 counts of 500 cells per μL or less cost $290 per DALY averted. In concentrated epidemics, expanded access for key populations was also cost effective. Our estimates suggest that earlier eligibility for antiretroviral therapy is very cost effective in low-income and middle-income settings, although these estimates should be revisited when more data become available. Scaling up antiretroviral therapy through earlier eligibility and expanded coverage should be considered alongside other high-priority health interventions competing for health budgets. Bill & Melinda Gates Foundation, WHO.


Dual promoter usage as regulatory mechanism of let-7c expression in leukemic and solid tumors.
Let-7c, an intronic microRNA (miRNA) embedded in the long non-coding gene LINC00478, can act as a tumor suppressor by targeting oncogenes. Previous studies indicated that in acute promyelocytic leukemia (APL), a subtype of acute myelogenous leukemia (AML) bearing the leukemia promoting PML/RARα fusion protein, let-7c expression seems to be controlled by the host gene promoter, in which canonical Retinoic Acid Responsive Elements (RAREs) are bound by PML/RARα in an all transretinoic acid (ATRA)-sensitive manner. Here, let-7c transcriptional regulation was further investigated and a novel intronic promoter upstream of the pre-miRNA was identified. This new promoter has transcriptional activity strongly indicating that at least two promoters need to be considered for let-7c transcription: the distal host gene and the proximal intronic promoter. Therefore, epigenetic modifying enzymes and histone acetylation and methylation status were analyzed on both let-7c promoters. It was demonstrated that ATRA treatment leads to let-7c upregulation inducing a more open chromatin conformation of the host gene promoter, with an enrichment of epigenetic marks that correlate with a more active transcriptional state. Conversely, the epigenetic marks on the intronic promoter are not significantly affected by ATRA treatment. Interestingly, in solid tumors such as prostate and lung adenocarcinoma it was found that both host and intronic promoters are functional. These data suggest that while the host gene promoter may control let-7c expression in AML, in a nonleukemic tumor context instead the intronic promoter contributes or preferentially regulates let-7c transcription. Alternative promoter usage represents a regulatory mechanism of let-7c expression in different tissues. Mol Cancer Res; 12(6); 878-89. ©2014 AACR.


Regulation of proline metabolism in mycobacteria and its role in carbon metabolism under hypoxia.
Genes with a role in proline metabolism are strongly expressed when mycobacterial cells are exposed to nutrient starvation and hypoxia. Here we show that proline metabolism in mycobacteria is mediated by the monofunctional enzymes Δ(1) -pyrroline-5-carboxylate dehydrogenase (PruA) and proline dehydrogenase (PruB). Proline metabolism was controlled by a unique membrane-associated DNA-binding protein PruC. Under hypoxia, addition of proline led to higher biomass production than in the absence of proline despite excess carbon and nitrogen. To identify the mechanism responsible for this enhanced growth, microarray analysis of wild-type Mycobacterium smegmatis versus pruC mutant was performed. Expression of the DNA repair machinery and glyoxalases was increased in the pruC mutant. Glyoxalases are proposed to degrade methylglyoxal, a toxic metabolite produced by various bacteria due to an imbalance in intermediary metabolism, suggesting the pruC mutant was under methylglyoxal stress. Consistent with this notion, pruB and pruC mutants were hypersensitive to methylglyoxal. Δ(1) -pyrroline-5-carboxylate is reported to react with methylglyoxal to form non-toxic 2-acetyl-1-pyrroline, thus providing a link between proline metabolism and methylglyoxal detoxification. In support of this mechanism, we show that proline metabolism protects mycobacterial cells from methylglyoxal toxicity and that functional proline dehydrogenase, but not Δ(1) -pyrroline-5-carboxylate dehydrogenase, is essential for this protective effect.


Sequencing the CYP2D6 gene: from variant allele discovery to clinical pharmacogenetic testing.
CYP2D6 is one of the most studied enzymes in the field of pharmacogenetics. The CYP2D6 gene is highly polymorphic with over 100 catalogued star (*) alleles, and clinical CYP2D6 testing is increasingly accessible and supported by practice guidelines. However, the degree of variation at the CYP2D6 locus and homology with its pseudogenes make interrogating CYP2D6 by short-read sequencing challenging. Moreover, accurate prediction of CYP2D6 metabolizer status necessitates analysis of duplicated alleles when an increased copy number is detected. These challenges have recently been overcome by long-read CYP2D6 sequencing; however, such platforms are not widely available. This review highlights the genomic complexities of CYP2D6, current sequencing methods and the evolution of CYP2D6 from allele discovery to clinical pharmacogenetic testing.


SU-E-I-100: Feasibility Study of Gamma-Ray Medical Radiography.
The purpose of this research is to study an alternative technique to conventional x-ray radiography that requires less patient radiation dose, less cost, portable, requires less maintenance, and less power consumption. This research explores the feasibility of using gamma-ray radiography in medical imaging. Gamma-ray medical radiography has the potential to provide alternative diagnostic medical information to X-ray radiography. Approximately one Ci of Am-241 radioactive source which emits monoenergetic 59.5 KeV gamma rays was used in this study. Several factors that influence this feasibility were studied. These were the radiation source uniformity, image uniformity, image quality parameters such as contrast, noise, and spatial resolution. In addition, visual assessment of several human phantom gamma-ray and x-ray images were conducted. The images were recorded on computed radiography image receptors and displayed on a standard monitor. The radioactive source provided a relatively uniform radiation exposure and uniform images. Image noise was mainly dependent on the exposure time and the source size. Although the contrast depended on the window and level setting, it was also dependent on the exposure time and the source size. Spatial resolution was dependent on the source size and the magnification. The generated gamma-ray images were of lower quality than the X-ray images which was mainly due to the low radioactivity used. However, the gamma-ray images displayed most of the main structures contained in the humanoid phantoms. Thisresearch explored the feasibility of using gamma-ray radiography in medical imaging and showed that gamma-ray medical radiography has the potential to provide alternative diagnostic medical information to X-ray radiography. Finally, this research also paves the way for the usage and production of high radioactive Am-241 source that will show high quality medical gamma-ray radiography is feasible.


Anaphe venata larva extract-induced purposeless chewing in rats: the role of cholinergic, GABAergic and opioid systems.
Seasonal ataxia was reported in humans following the consumption ofAnaphe venata larva as protein supplement in diet and altered motor function in rodents when the extract was administered intraperitoneally. In this study we investigated the effect of the crude aqueous and Phosphate Buffer Saline (PBS) extracts of this larva on altered spontaneous rat behavior in a novel environment particularly chewing behaviour, with a view to determine the mechanism(s) involved in these behavioural alteration. Animals were randomly assigned into four groups (n = 6-12 per group) and graded doses of aqueous and PBS extracts (100-400 mg/kg) were administered dissolved in saline intraperitoneally (i.p.) to each animal in the experimental groups. The control group received an equivalent volume of saline. Behavioral scores were recorded for a period of 30 minutes after the administration of saline or extract. The role of various receptors in the extract induced chewing was evaluated using known receptor agonist/antagonists. Results revealed a significant increase in purposeless chewing (F (7, 95) = 7.85; p <0.05) by the aqueous extract compared to saline control at all dose levels, which was significantly attenuated by scopolamine (3 mg/kg, i.p) and thiamine (1 mg/kg, i.p) respectively (p <0.05); while flumazenil (2 mg/kg, i.p) and naloxone (2.5 mg/kg, i.p) did not alter the induced purposeless chewing behaviour. Also, administration of PBS induced a significant (F (7, 95) = 6.11; p <0.05) increase in chewing behaviour but only at 400 mg/kg dose level which was attenuated by scopolamine (3 mg/kg, i.p); while flumazenil (2 mg/kg, i.p), naloxone (2.5 mg/kg, i.p), and thiamine (1 mg/kg, i.p) potentiated purposeless chewing behaviour respectively. It may therefore be concluded from this study that Anaphe extract-induced purposeless chewing behaviour in rat is mediated via the activation of cholinergic neurotransmission which is modulated by GABAergic and opioid receptor systems.


Reduced acquisition and reactivation of human papillomavirus infections among older women treated with cryotherapy: results from a randomized trial in South Africa.
Treatment of women for high-grade cervical cancer precursors frequently results in clearance of the associated high-risk human papillomavirus (hrHPV) infection but the role of treatment among women without hrHPV is unknown. We investigated whether cervical cryotherapy reduces newly detected hrHPV infections among HIV-positive and HIV-negative women who were hrHPV negative when treated. The impact of cryotherapy on newly detected hrHPV infections was examined among 612 women of known HIV serostatus, aged 35 to 65 years, who were negative for hrHPV DNA, and randomized to either undergo cryotherapy (n = 309) or not (n = 303). All women underwent repeat hrHPV DNA testing 6, 12, 24, and 36 months later. Among 540 HIV-negative women, cryotherapy was associated with a significant reduction in newly detected hrHPV infections. Women in the cryotherapy group were 55% less likely to have newly detected hrHPV than women in the control group (95% CI 0.28 to 0.71). This association was independent of the influence of changes in sexual behaviors following therapy (adjusted hazards ratio (HR) = 0.49, 95% CI 0.29 to 0.81). Among 72 HIV-positive women, similar reductions were not observed (HR = 1.10, 95% CI 0.53 to 2.29). Cervical cryotherapy significantly reduced newly detected hrHPV infections among HIV-negative, but not HIV-positive women. These results raise intriguing questions about immunological responses and biological mechanisms underlying the apparent prophylactic benefits of cryotherapy.


Evidence for a SAL1-PAP chloroplast retrograde pathway that functions in drought and high light signaling in Arabidopsis.
Compartmentation of the eukaryotic cell requires a complex set of subcellular messages, including multiple retrograde signals from the chloroplast and mitochondria to the nucleus, to regulate gene expression. Here, we propose that one such signal is a phosphonucleotide (3'-phosphoadenosine 5'-phosphate [PAP]), which accumulates in Arabidopsis thaliana in response to drought and high light (HL) stress and that the enzyme SAL1 regulates its levels by dephosphorylating PAP to AMP. SAL1 accumulates in chloroplasts and mitochondria but not in the cytosol. sal1 mutants accumulate 20-fold more PAP without a marked change in inositol phosphate levels, demonstrating that PAP is a primary in vivo substrate. Significantly, transgenic targeting of SAL1 to either the nucleus or chloroplast of sal1 mutants lowers the total PAP levels and expression of the HL-inducible ASCORBATE PEROXIDASE2 gene. This indicates that PAP must be able to move between cellular compartments. The mode of action for PAP could be inhibition of 5' to 3' exoribonucleases (XRNs), as SAL1 and the nuclear XRNs modulate the expression of a similar subset of HL and drought-inducible genes, sal1 mutants accumulate XRN substrates, and PAP can inhibit yeast (Saccharomyces cerevisiae) XRNs. We propose a SAL1-PAP retrograde pathway that can alter nuclear gene expression during HL and drought stress.


Blood flow responses to intra-arterial acetylcholine in man: effects of basal flow and conduit vessel length.
1. Acetylcholine relaxes human resistance vessels and releases nitric oxide and other factors from the endothelium. Comparison of responses to acetylcholine with those to an endothelium-independent vasodilator (such as nitroprusside) forms the basis of the so-called acetylcholine test of endothelial function. However, when this test is applied in vivo by intraarterial infusion, the metabolic instability of acetylcholine may result in differential responses to these drugs arising from anatomical rather than functional differences. 2. Vasodilator responses to brachial artery infusions of acetylcholine (41 and 83 nmol/min) and sodium nitroprusside (11 and 38 nmol/min) were measured in 30 healthy human subjects using venous occlusion plethysmography. 3. Responses to acetylcholine showed a greater dependence on resting blood flow (P < 0.05) and on forearm length (P < 0.05) than those to sodium nitroprusside (results predicted by a simple blood flow model). 4. Correction for forearm length abolished an apparent difference of 59% between responses to acetylcholine in men and women. Conduit vessel geometry and resting blood flow influence the acetylcholine test of endothelial function.


DREAM is reduced in synovial fibroblasts of patients with chronic arthritic pain: is it a suitable target for peripheral pain management?
The endogenous pain-relieving system depends in part on the regulation of nociceptive signals through binding of opioids to the corresponding opioid receptor. Interfering with the trans-repression effect of downstream regulatory element antagonist modulator (DREAM) on the transcription of the opioid dynorphin-encoding prodynorphin (pdyn) gene might enhance pain relief in the periphery. Expression levels were measured in osteoarthritis (OA) synovial fibroblast-like cells (SFLCs) (n = 8) and in peripheral blood mononuclear cells (PBMCs) from OA patients (n = 53) and healthy controls (n = 26) by real-time polymerase chain reaction. Lysed OA SFLCs were analyzed by immunoprecipitation. Translation of DREAM mRNA was inhibited by small interfering RNAs (siRNAs). Expressions of DREAM, pdyn, and c-fos mRNAs were measured at 24, 48, and 72 hours after transfection. The expression of DREAM mRNA was shown in both healthy and OA SFLCs as well as PBMCs. Inhibiting transcription using siRNAs led to a marked reduction in DREAM expression after 24, 48, and 72 hours. However, no significant changes in c-fos and pdyn expression occurred. In addition, DREAM mRNA expression was significantly reduced in OA patients with chronic pain (pain intensity as measured by a visual analog scale scale of greater than 40), but no pdyn expression was detectable. To our knowledge, this is the first report showing the expression of DREAM in SFLCs and PBMCs on the mRNA level. However, DREAM protein was not detectable. Since repression of pdyn transcription persists after inhibiting DREAM translation, DREAM appears to play no functional role in the kappa opioid receptor system in OA SFLCs. Therefore, our data suggest that DREAM appears not to qualify as a target in peripheral pain management.


The effect of ischemic time and temperature on lung preservation in a simple ex vivo rabbit model used for functional assessment.
Despite 25 years of research in lung transplantation, little progress has been made in methods for improving lung preservation. To evaluate many factors that may affect lung function after preservation, we have developed a simple, reliable, and inexpensive animal model. This consists of an isolated rabbit lung preparation perfused with blood and ventilated with a closed-circuit system. Heart-lung blocks were harvested and the left lung was assessed after ligation of the right pulmonary artery and right main-stem bronchus. On completion of a storage period, the left lung was ventilated and perfused with fresh rabbit venous blood at a rate of 40 ml/min for 10 minutes. Assessment of lung function included gas analysis of infused and effluent blood, oxygen uptake, mean pulmonary artery perfusion pressure, and mean airway pressure. A control group and six preservation groups were evaluated, each with different storage temperatures (38 degrees C, 34 degrees C, 23 degrees C, 15 degrees C, 10 degrees C, and 4 degrees C). For each temperature, ischemic periods ranging from 1 to 30 hours were studied. In the control group, the lungs were assessed immediately after being harvested. In the preservation groups, the lungs were kept partially inflated, stored at a predetermined temperature by immersion, and later assessed after variable ischemic periods. Our studies demonstrated the following: (1) The degree of impaired lung function produced by ischemia is reflected by a decrease in oxygen uptake and in oxygen tension of the effluent pulmonary venous blood and an increase in pulmonary artery perfusion pressure; (2) hypothermia improves ischemic tolerance; (3) preservation of lung at 10 degrees C is superior to preservation at 15 degrees C and 4 degrees C. This screening model has allowed evaluation of independent multiple factors and methods pertinent to lung preservation and enables one to assess lung function reliably and rapidly.


Proteins differentially expressed in human beta-cells-enriched pancreatic islet cultures and human insulinomas.
In view of the great demand for human beta-cells for physiological and medical studies, we generated cell lines derived from human insulinomas which secrete insulin, C-peptide and express neuroendocrine and islet markers. In this study, we set out to characterize their proteomes, comparing them to those of primary beta-cells using DIGE followed by MS. The results were validated by Western blotting. An average of 1800 spots was detected with less than 1% exhibiting differential abundance. Proteins more abundant in human islets, such as Caldesmon, are involved in the regulation of cell contractility, adhesion dependent signaling, and cytoskeletal organization. In contrast, almost all proteins more abundant in insulinoma cells, such as MAGE2, were first described here and could be related to cell survival and resistance to chemotherapy. Our proteomic data provides, for the first time, a molecular snapshot of the orchestrated changes in expression of proteins involved in key processes which could be correlated with the altered phenotype of human beta-cells. Collectively our observations prompt research towards the establishment of bioengineered human beta-cells providing a new and needed source of cultured human beta-cells for beta-cell research, along with the development of new therapeutic strategies for detection, characterization and treatment of insulinomas.


Laparoscopic nephrectomy for emphysematous pyelonephritis.
Emphysematous pyelonephritis is a serious suppurative infection of the renal and extrarenal tissues. Controversies exist surrounding the appropriate management of this life-threatening condition with advocates for both medical management alone and percutaneous drainage combined with nephrectomy. The laparoscopic approach for nephrectomy is quickly becoming the standard of care for benign disease of the kidney and low-stage renal cancers. We report a case of a 60-year-old diabetic male undergoing successful laparoscopic nephrectomy for right emphysematous pyelonephritis, and our procedural technique. To our knowledge, this is the first report of laparoscopic nephrectomy in the setting of emphysematous pyelonephritis. Carefully selected patients with emphysematous pyelonephritis may benefit from the advantages of laparoscopic nephrectomy in centers with sufficient laparoscopic experience.


Selective importance of the rat anterior thalamic nuclei for configural learning involving distal spatial cues.
To test potential parallels between hippocampal and anterior thalamic function, rats with anterior thalamic lesions were trained on a series of biconditional learning tasks. The anterior thalamic lesions did not disrupt learning two biconditional associations in operant chambers where a specific auditory stimulus (tone or click) had a differential outcome depending on whether it was paired with a particular visual context (spot or checkered wall-paper) or a particular thermal context (warm or cool). Likewise, rats with anterior thalamic lesions successfully learnt a biconditional task when they were reinforced for digging in one of two distinct cups (containing either beads or shredded paper), depending on the particular appearance of the local context on which the cup was placed (one of two textured floors). In contrast, the same rats were severely impaired at learning the biconditional rule to select a specific cup when in a particular location within the test room. Place learning was then tested with a series of go/no-go discriminations. Rats with anterior thalamic nuclei lesions could learn to discriminate between two locations when they were approached from a constant direction. They could not, however, use this acquired location information to solve a subsequent spatial biconditional task where those same places dictated the correct choice of digging cup. Anterior thalamic lesions produced a selective, but severe, biconditional learning deficit when the task incorporated distal spatial cues. This deficit mirrors that seen in rats with hippocampal lesions, so extending potential interdependencies between the two sites.


Minocycline prevents morphine-induced apoptosis in rat cerebral cortex and lumbar spinal cord: a possible mechanism for attenuating morphine tolerance.
Tolerance to the chronic administration of opioids such as morphine reduces the utility of these drugs in pain management. Despite significant investigation, the precise cellular mechanisms underlying opioid tolerance and dependence remain elusive. It has been indicated that tolerance to the analgesic effect of morphine is associated with apoptosis in the central nervous system. The aim of this study was to examine the effects of the intracerebroventricular (icv) administration of minocycline (a second-generation tetracycline) on morphine-induced apoptosis in the cerebral cortex and lumbar spinal cord of rats after morphine-induced tolerance. Different groups of rats received either morphine (ip) and distilled water (icv) or morphine and different doses of minocycline (icv) or minocycline alone once per day. The terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method was used to analyze apoptosis. The anti-apoptotic factors, Bcl-2 and HSP 70 and the pro-apoptotic element caspase-3 were evaluated by immunoblotting. The results indicated that minocycline attenuated the number of apoptotic cells in both the cerebral cortex and lumbar spinal cord. Immunoblotting findings showed that the amounts of anti-apoptotic agents (Bcl-2 and HSP 70) were greater in the treatment groups than in the controls in both regions. Although minocycline did not change the level of caspase-3 at the doses used with morphine but the minocycline treated rats showed a significantly lower increase in caspase-3 activity than did in the control. In conclusion, minocycline decreased the number of TUNEL-positive cells and increased the amount of anti-apoptotic factors (Bcl-2 and HSP 70), but did not change the caspase-3 content.


Using restraint with nursing home residents: a qualitative study of nursing staff perceptions and decision-making.
The study reported in this paper applied a qualitative and interpretative approach to nursing staff perceptions of the use of restraint with elderly nursing home residents, and into nurses' decision-making on restraint use. The data were collected using unstructured interviews with a purposive sample of 20 trained and untrained nursing staff from two Swiss nursing homes. Data analysis was based on Colaizzi's phenomenological method. Three main themes were extracted from the data: (1) understanding the term restraint, (2) situations in which the decision to apply restraint is considered justified and (3) situations in which nursing staff are uncertain about the use of restraint. The underlying bases with respect to decision-making were: understanding restraint, the rights and responsibilities of both residents and staff, and the duties of staff. Staff members were ambiguous in their understanding of restraint and they showed positive as well as confused attitudes towards its use. Their behaviour was defensive and protective rather than challenging. Further research is required on what is meant by safety in care of the elderly nursing today. In nursing practice, as far as issues of restraint are concerned, greater attention should be devoted to the relationship between elderly residents' self-determination and responsibility for their actions.


Schwann cell galactocerebroside induced by derivatives of adenosine 3',5'-monophosphate.
Indirect immunofluorescence was used to show the presence of galactocerebroside (galC), a lipid found in myelin, on the surface of about half of the Schwann cells isolated from neonatal rat sciatic nerves and cultured for 1 day without neurons. By day 4 in vitro, the Schwann cells had all lost their surface galC. Three days after beginning treatment with 10(-3) molar 8-bromo-adenosine 3',5'-monophosphate (8-bromo cyclic AMP) or N6,O2'-dibutyryl adenosine 3',5'-monophosphate (dibutyryl cyclic AMP), galC reappeared on the Schwann cells, and 2 days later 48 percent of the cultured Schwann cells showed surface galC. Tritium from tritiated D-galactose was incorporated into galC by the 8-bromo cyclic AMP-and dibutyryl cyclic AMP-treated cultures at a rate 15 times the control rate.


Azoxymethane-induced fulminant hepatic failure in C57BL/6J mice: characterization of a new animal model.
Without transplantation, approximately 50-90% of all patients with fulminant hepatic failure (FHF) die. This poor outcome is due in part to the absence of an appropriate animal model, which would allow for a greater understanding of the pathophysiology of this syndrome. Given the reports of liver injury in humans and livestock fed cycad palm nuts on the island of Guam, we hypothesized that the active ingredient azoxymethane (AOM) could cause FHF. We therefore evaluated AOM in C57BL/6J mice. Histologically, we observed microvesicular steatosis 2 h, sinusoidal dilatation 4 h, and centrilobular necrosis 20 h after AOM administration, and transmission electron microscopy showed that this agent causes mitochondrial injury. FHF was associated with all four stages of encephalopathy, as well as by a prodromal period of decreased eating and drinking lasting approximately 15 h before the development of stage I encephalopathy (i.e., loss of scatter reflex). Late encephalopathy was associated with increased arterial ammonia, decreased serum glucose, and evidence of brain edema (astrocyte swelling). We show that AOM-induced FHF is highly reproducible, without evidence of lot-to-lot variability, and is dose dependent. These findings therefore suggest that AOM is an excellent agent for the study of FHF, as well as indicate that Guamanian FHF may be due to AOM found in unwashed cycad palm nuts.


Removing financial barriers to access reproductive, maternal and newborn health services: the challenges and policy implications for human resources for health.
The last decade has seen widespread retreat from user fees with the intention to reduce financial constraints to users in accessing health care and in particular improving access to reproductive, maternal and newborn health services. This has had important benefits in reducing financial barriers to access in a number of settings. If the policies work as intended, service utilization rates increase. However this increases workloads for health staff and at the same time, the loss of user fee revenues can imply that health workers lose bonuses or allowances, or that it becomes more difficult to ensure uninterrupted supplies of health care inputs.This research aimed to assess how policies reducing demand-side barriers to access to health care have affected service delivery with a particular focus on human resources for health. We undertook case studies in five countries (Ghana, Nepal, Sierra Leone, Zambia and Zimbabwe). In each we reviewed financing and HRH policies, considered the impact financing policy change had made on health service utilization rates, analysed the distribution of health staff and their actual and potential workloads, and compared remuneration terms in the public sectors. We question a number of common assumptions about the financing and human resource inter-relationships. The impact of fee removal on utilization levels is mostly not sustained or supported by all the evidence. Shortages of human resources for health at the national level are not universal; maldistribution within countries is the greater problem. Low salaries are not universal; most of the countries pay health workers well by national benchmarks. The interconnectedness between user fee policy and HRH situations proves difficult to assess. Many policies have been changing over the relevant period, some clearly and others possibly in response to problems identified associated with financing policy change. Other relevant variables have also changed.However, as is now well-recognised in the user fee literature, co-ordination of health financing and human resource policies is essential. This appears less well recognised in the human resources literature. This coordination involves considering user charges, resource availability at health facility level, health worker pay, terms and conditions, and recruitment in tandem. All these policies need to be effectively monitored in their processes as well as outcomes, but sufficient data are not collected for this purpose.


The stroke prognosis instrument II (SPI-II) : A clinical prediction instrument for patients with transient ischemia and nondisabling ischemic stroke.
In 1991 we developed the Stroke Prognosis Instrument (SPI-I) to stratify patients with transient ischemic attack or ischemic stroke by prognosis for stroke or death in 2 years. In this article we validate and improve SPI-I (creating SPI-II). To validate SPI-I, we applied it to 4 test cohorts and calculated pooled outcome rates. To create SPI-II, we incorporated new predictive variables identified in 1 of the test cohorts and validated it in the other 3 cohorts. For SPI-I, pooled rates (all 4 test cohorts) of stroke or death within 2 years in risk groups I, II, and III were 9%, 17%, and 24%, respectively (P<0.01, log-rank test). SPI-II was created by adding congestive heart failure and prior stroke to SPI-I. Each patient's risk group was determined by the total score for 7 factors: congestive heart failure (3 points); diabetes (3 points); prior stroke (3 points); age >70 years (2 points); stroke for the index event (not transient ischemic attack) (2 points); hypertension (1 point); and coronary artery disease (1 point). Risk groups I, II, and III comprised patients with 0 to 3, 4 to 7, and 8 to 15 points, respectively. For SPI-I, pooled rates (3 cohorts excluding the SPI-II development cohort) of stroke or death within 2 years in risk groups I, II, and III were 9%, 17%, and 23%, respectively. For SPI-II, pooled rates were 10%, 19%, and 31%, respectively. In receiver operator characteristic analysis, the area under the curve was 0.59 (95% CI, 0.57 to 0.60) for SPI-I and 0.63 (95% CI, 0.62 to 0.65) for SPI-II, confirming the better performance of the latter. Compared with SPI-I, SPI-II achieves greater discrimination in outcome rates among risk groups. SPI-II is ready for use in research design and may have a role in patient counseling.


Delays in diagnosis and treatment of pulmonary tuberculosis in India: a systematic review.
To systematically review Indian literature on delays in tuberculosis (TB) diagnosis and treatment. We searched multiple sources for studies on delays in patients with pulmonary TB and those with chest symptoms. Studies were included if numeric data on any delay were reported. Patient delay was defined as the interval between onset of symptoms and the patient's first contact with a health care provider. Diagnostic delay was defined as the interval between the first consultation with a health care provider and diagnosis. Treatment delay was defined as the interval between diagnosis and initiation of anti-tuberculosis treatment. Total delay was defined as time interval from the onset of symptoms until treatment initiation. Among 541 potential citations identified, 23 studies met the inclusion criteria. Included studies used a variety of definitions for onset of symptoms and delays. Median estimates of patient, diagnostic and treatment delay were respectively 18.4 (IQR 14.3-27.0), 31.0 (IQR 24.5-35.4) and 2.5 days (IQR 1.9-3.6) for patients with TB and those with chest symptoms combined. The median total delay was 55.3 days (IQR 46.5-61.5). About 48% of all patients first consulted private providers; an average of 2.7 health care providers were consulted before diagnosis. Number and type of provider first consulted were the most important risk factors for delay. These findings underscore the need to develop novel strategies for reducing patient and diagnostic delays and engaging first-contact health care providers.


eIF5A-PEAK1 Signaling Regulates YAP1/TAZ Protein Expression and Pancreatic Cancer Cell Growth.
In pancreatic ductal adenocarcinoma (PDAC), mutant KRAS stimulates the translation initiation factor eIF5A and upregulates the focal adhesion kinase PEAK1, which transmits integrin and growth factor signals mediated by the tumor microenvironment. Although eIF5A-PEAK1 signaling contributes to multiple aggressive cancer cell phenotypes, the downstream signaling processes that mediate these responses are uncharacterized. Through proteomics and informatic analyses of PEAK1-depleted PDAC cells, we defined protein translation, cytoskeleton organization, and cell-cycle regulatory pathways as major pathways controlled by PEAK1. Biochemical and functional studies revealed that the transcription factors YAP1 and TAZ are key targets of eIF5A-PEAK1 signaling. YAP1/TAZ coimmunoprecipitated with PEAK1. Interfering with eIF5A-PEAK1 signaling in PDAC cells inhibited YAP/TAZ protein expression, decreasing expression of stem cell-associated transcription factors (STF) including Oct4, Nanog, c-Myc, and TEAD, thereby decreasing three-dimensional (3D) tumor sphere growth. Conversely, amplified eIF5A-PEAK1 signaling increased YAP1/TAZ expression, increasing expression of STF and enhancing 3D tumor sphere growth. Informatic interrogation of mRNA sequence databases revealed upregulation of the eIF5A-PEAK1-YAP1-TEAD signaling module in PDAC patients. Taken together, our findings indicate that eIF5A-PEAK1-YAP signaling contributes to PDAC development by regulating an STF program associated with increased tumorigenicity. Cancer Res; 77(8); 1997-2007. ©2017 AACR.


Pulmonary dysfunction in cystic fibrosis is associated with oxidative stress.
The aim of this study was to determine whether a relationship exists between the circulating concentration of antioxidants, or markers of oxidative stress, and pulmonary function in cystic fibrosis patients. Plasma was obtained from 34 patients attending a cystic fibrosis clinic. Oxidative stress was investigated by measuring the concentrations of circulating lipid hydroperoxides and malondialdehyde (lipid peroxidation) and protein carbonyls (protein oxidation). Antioxidant status was determined from the plasma concentrations of alpha-tocopherol, ascorbic acid, uric acid and total sulphydryls. Forced vital capacity (FVC), forced expiratory volume in one second (FEV1) and forced mid-expiratory flow (FEF25-75) were measured in 25 of the subjects by spirometry, and expressed as percentage predicted for normal height, weight and age. Lung function decreased significantly with age and was associated with decreased plasma alpha-tocopherol, ascorbic acid and sulphydryl concentrations. The reduction in pulmonary function correlated with elevated plasma malondialdehyde, but not with lipid hydroperoxide or protein carbonyl concentrations. Patients with severe lung dysfunction (FEV1 < 50% predicted) had higher plasma concentrations of lipid hydroperoxides than those with mild-to-moderate lung dysfunction (FEV1 > 50% pred). This study provides evidence that cystic fibrosis patients have inadequate antioxidant defences to cope with the elevated oxidative stress that they regularly experience. We believe that recurring oxidative lung injury contributes to the decline in pulmonary function in these patients.


Energy Drink and Alcohol mixed Energy Drink use among high school adolescents: Association with risk taking behavior, social characteristics.
The aims of the study were to: a) examine the prevalence of energy drink (ED) and alcohol mixed with energy drink (AmED) consumption; b) investigate the relationships between ED and AmED with alcohol, binge drinking and drugs accounting for at risk behaviors among a representative sample of Italian adolescents. A representative sample of 30,588 Italian high school students, aged 15-19years, was studied. Binary and multivariate logistic regression analyses were performed to determine the independent association of the potential predictors' characteristics with the ED and AmED drinking during the last year. Respectively 41.4% and 23.2% of respondents reported drinking EDs and AmEDs in the last year. Multivariate analysis revealed that consumption of EDs and AmEDs during the last year were significantly associated with daily smoking, binge drinking, use of cannabis and other psychotropic drugs. Among life habits and risky behaviors the following were positively associated: going out with friends for fun, participating in sports, experiencing physical fights/accidents or injury, engaging in sexual intercourse without protection and being involved in accidents while driving. This study demonstrates the popularity of ED and AmED consumption among the Italian school population aged 15-19years old: 4 out of 10 students consumed EDs in the last year and 2 out of 10 AmED. Multivariate analysis highlighted the association with illicit drug consumption and harming behaviors, confirming that consumption of EDs and AmEDs is a compelling issue especially during adolescence, as it can effect health as well as risk taking behaviors.


Optic nerve sheath diameter measurement: a means of detecting raised ICP in adult traumatic and non-traumatic neurosurgical patients.
Bedside ultrasound measurement of optic nerve sheath diameter (ONSD) is emerging as a non-invasive technique to evaluate and predict raised intracranial pressure (ICP). It has been shown in previous literature that ONSD measurement has good correlation with surrogate findings of raised ICP such as clinical and radiological findings suggestive of raised ICP. The objective of the study is to find a correlation between sonographic measurements of ONSD value with ICP value measured via the gold standard invasive intracranial ICP catheter, and to find the cut-off value of ONSD measurement in predicting raised ICP, along with its sensitivity and specificity value. A prospective observational study was performed using convenience sample of 41 adult neurosurgical patients treated in neurosurgical intensive care unit with invasive intracranial pressure monitoring placed in-situ as part of their clinical care. Portable SonoSite ultrasound machine with 7 MHz linear probe were used to measure optic nerve sheath diameter using the standard technique. Simultaneous ICP readings were obtained directly from the invasive monitoring. Seventy-five measurements were performed on 41 patients. The non-parametric Spearman correlation test revealed a significant correlation at the 0.01 level between the ICP and ONSD value, with correlation coefficient of 0.820. The receiver operating characteristic curve generated an area under the curve with the value of 0.964, and with standard error of 0.22. From the receiver operating characteristic curve, we found that the ONSD value of 5.205 mm is 95.8% sensitive and 80.4% specific in detecting raised ICP. ONSD value of 5.205 is sensitive and specific in detecting raised ICP. Bedside ultrasound measurement of ONSD is readily learned, and is reproducible and reliable in predicting raised ICP. This non-invasive technique can be a useful adjunct to the current invasive intracranial catheter monitoring, and has wide potential clinical applications in district hospitals, emergency departments and intensive care units.


In Vitro Activity of Tigecycline and Colistin against clinical isolates of Acinetobacter baumannii in Hospitals in Tehran and Bandar-Abbas, Iran.
The Acinetobacter species, particularly A. baumannii, has emerged as one of the main causes of nosocomial infections in recent years. The high prevalence of drug resistance in A. baumannii limits the therapeutic options for treating infections caused by these bacteria. The objective of this study was to determine the in vitro activity of Tigecycline and Colistin against clinical isolates of A. baumannii in Tehran and Bandar Abbas, Iran. This study was conducted from March 2009 to November 2010 at three hospitals in Tehran and Bandar Abbas, Iran, using 165 Acinetobacter species isolated from clinical specimens. All isolates were subjected to PCR to detect bla OXA-51-like genes that are unique to Acinetobacter baumannii. Isolates that gave a band for the bla OXA-51-like genes were identified as A. baumannii. Anti-microbial susceptibility tests were performed for Tigecycline, Colistin, and other antibiotics. Sensitivity rates to Colistin and Polymyxin-B were 100%. Resistance rates for Tigecycline were 4.2% in Tehran and 8.8% in Bandar-Abbas according to Jones criteria, whereas, according to U.S. FDA criteria, the resistance rates were 20.8% and 17.6%, respectively. New alternative drugs are needed for the treatment of drug resistant A. baumannii. Although Colistin appears to be a good choice, adverse reactions have limited its usage. Tigecycline is effective against A. baumannii isolates, and it shows promise for solving the problem.


Selective activation of the B natriuretic peptide receptor by C-type natriuretic peptide (CNP).
The natriuretic peptides are hormones that can stimulate natriuretic, diuretic, and vasorelaxant activity in vivo, presumably through the activation of two known cell surface receptor guanylyl cyclases (ANPR-A and ANPR-B). Although atrial natriuretic peptide (ANP) and, to a lesser extent, brain natriuretic peptide (BNP) are efficient activators of the ANPR-A guanylyl cyclase, neither hormone can significantly stimulate ANPR-B. A member of this hormone family, C-type natriuretic peptide (CNP), potently and selectively activated the human ANPR-B guanylyl cyclase. CNP does not increase guanosine 3',5'-monophosphate accumulation in cells expressing human ANPR-A. The affinity of CNP for ANPR-B is 50- or 500-fold higher than ANP or BNP, respectively. This ligand-receptor pair may be involved in the regulation of fluid homeostasis by the central nervous system.


Control of Emi2 activity and stability through Mos-mediated recruitment of PP2A.
Before fertilization, vertebrate eggs are arrested in meiosis II by cytostatic factor (CSF), which holds the anaphase-promoting complex (APC) in an inactive state. It was recently reported that Mos, an integral component of CSF, acts in part by promoting the Rsk-mediated phosphorylation of the APC inhibitor Emi2/Erp1. We report here that Rsk phosphorylation of Emi2 promotes its interaction with the protein phosphatase PP2A. Emi2 residues adjacent to the Rsk phosphorylation site were important for PP2A binding. An Emi2 mutant that retained Rsk phosphorylation but lacked PP2A binding could not be modulated by Mos. PP2A bound to Emi2 acted on two distinct clusters of sites phosphorylated by Cdc2, one responsible for modulating its stability during CSF arrest and one that controls binding to the APC. These findings provide a molecular mechanism for Mos action in promoting CSF arrest and also define an unusual mechanism, whereby protein phosphorylation recruits a phosphatase for dephosphorylation of distinct sites phosphorylated by another kinase.


Influence of birth weight on internalizing traits modulated by serotonergic genes.
Fetal growth predicts childhood behavioral problems associated with brain serotonergic systems. We hypothesized that allelic variations in genes involved in serotonergic function would moderate associations between birth weight (BW) and internalizing traits in childhood. The Child Behavior Checklist was administered to 545 healthy Singaporean children at 8 to 12 years. BW, corrected for gestational age, and candidate single-nucleotide polymorphisms (SNPs) in the TPH2, HTR2A, and SCL6A4 genes were investigated. There was no significant main effect of BW on internalizing T scores (F = 1.08; P = .36). After multiple corrections, significant main effects on internalizing T scores were found for HTR2A rs2296972 (adjusted: F = 2.85; P = .019) and HTR2A rs6313 (adjusted: F = 5.91; P = .0002). Significant interactions were found between BW and SNPs for the TPH2 gene (rs2171363: P = .008; rs7305115: P = .007) and the HTR2A gene (rs2770304: P = .001; rs6313: P = .026) for internalizing T scores. The CC genotype of TPH2 rs2171363, GG genotype of TPH2 rs7305115, CC genotype of HTR2A rs2770304, and CC genotype of HTR2A rs6313 were associated with reduced internalizing scores for children born in the quartile above the midpoint. No significant main effects or interactions were found for SCL6A4 SNPs. These findings suggest that sequence variations in genes involved in serotonergic functions modulate relationships between BW and internalizing traits and might be candidates for plasticity mechanisms that determine individual differences in responses to environmental influences over the course of development.


Identity and diversity of blood meal hosts of biting midges (Diptera: Ceratopogonidae: Culicoides Latreille) in Denmark.
Host preference studies in haematophagous insects e.g. Culicoides biting midges are pivotal to assess transmission routes of vector-borne diseases and critical for the development of veterinary contingency plans to identify which species should be included due to their risk potential. Species of Culicoides have been found in almost all parts of the world and known to live in a variety of habitats. Several parasites and viruses are transmitted by Culicoides biting midges including Bluetongue virus and Schmallenberg virus. The aim of the present study was to determine the identity and diversity of blood meals taken from vertebrate hosts in wild-caught Culicoides biting midges near livestock farms. Biting midges were collected at weekly intervals for 20 weeks from May to October 2009 using light traps at four collection sites on the island Sealand, Denmark. Blood-fed female biting midges were sorted and head and wings were removed for morphological species identification. The thoraxes and abdomens including the blood meals of the individual females were subsequently subjected to DNA isolation. The molecular marker cytochrome oxidase I (COI barcode) was applied to identify the species of the collected biting midges (GenBank accessions JQ683259-JQ683374). The blood meals were first screened with a species-specific cytochrome b primer pair for cow and if negative with a universal cytochrome b primer pair followed by sequencing to identify mammal or avian blood meal hosts. Twenty-four species of biting midges were identified from the four study sites. A total of 111,356 Culicoides biting midges were collected, of which 2,164 were blood-fed. Specimens of twenty species were identified with blood in their abdomens. Blood meal sources were successfully identified by DNA sequencing from 242 (76%) out of 320 Culicoides specimens. Eight species of mammals and seven species of birds were identified as blood meal hosts. The most common host species was the cow, which constituted 77% of the identified blood meals. The second most numerous host species was the common wood pigeon, which constituted 6% of the identified blood meals. Our results suggest that some Culicoides species are opportunistic and readily feed on a variety of mammals and birds, while others seems to be strictly mammalophilic or ornithophilic. Based on their number, dispersal potential and blood feeding behaviour, we conclude that Culicoides biting midges are potential vectors for many pathogens not yet introduced to Denmark.


Ultrastructural examination of the host inflammatory response within gills of netpen reared chinook salmon (Oncorhynchus tshawytscha) with Microsporidial Gill Disease.
The sequence of host changes following the rupture of spore-laden xenomas of the microsporidian Loma salmonae during Microsporidial Gill Disease of Salmon was deduced from ultrastructural examination of the gills of naturally infected, moribund, chinook salmon from a commercial aquaculture site. The gills contained many stages of parasite development suggesting fish were chronically exposed to the parasite. Intact xenomas were generally found beneath the endothelium in arteries and arterioles and were encapsulated by a layer of collagen containing fibroblasts sometimes joined by desmosomes. Xenoma dissolution was characterized by neutrophil infiltration and loss of the xenoma plasma membrane and encapsulation. The inflammatory responses associated with ruptured xenomas ranged from acute lesions, denoted by a marked neutrophil infiltration and vascular thrombosis, to chronic lesions with a macrophage-rich infiltrate variously accompanied by neovascularization and vascular remodelling. Dendritic-like cells and plasma cells were characteristic throughout. Basement membrane damage of the primary filament epithelium and subsequent transepithelial expulsion of spores were associated with severe inflammation. An unusual previously undescribed multifocal change, in which epithelial cells invaded deeply beyond the normal boundaries of the basement membrane, affected areas of gill filament epithelium with basement membrane damage. Some neutrophils that contained L. salmonae spores, or spore polar tube, displayed morphological changes that included irregular cell shape, cytoplasmic darkening associated with an abundance of free ribosomes, lysis of neighbouring cells, and type II nuclear clefts. Fusion of apparently intact neutrophils occurred in other areas of the lesion, where close contacts between neighbouring cells were established and in some areas plasma membrane fusion occurred. Closely associated neutrophils with intact plasma membranes were observed to contain type II nuclear clefts, abundant granules and rough endoplasmic reticulum. Other neutrophils in the lesion displayed type I nuclear pockets, which is suspected to be an early stage of apoptosis.


Local Submodularization for Binary Pairwise Energies.
Many computer vision problems require optimization of binary non-submodular energies. We propose a general optimization framework based on local submodular approximations (LSA). Unlike standard LP relaxation methods that linearize the whole energy globally, our approach iteratively approximates the energy locally. On the other hand, unlike standard local optimization methods (e.g., gradient descent or projection techniques) we use non-linear submodular approximations and optimize them without leaving the domain of integer solutions. We discuss two specific LSA algorithms based on trust region and auxiliary function principles, LSA-TR and LSA-AUX. The proposed methods obtain state-of-the-art results on a wide range of applications such as binary deconvolution, curvature regularization, inpainting, segmentation with repulsion and two types of shape priors. Finally, we discuss a move-making extension to the LSA-TR approach. While our paper is focused on pairwise energies, our ideas extend to higher-order problems. The code is available online.


Management of cholestatic disease in 2017.
Primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) are the most frequent chronic cholestatic liver diseases and serve as model diseases to discuss the management of cholestasis in 2017 in the lecture that is summarized in this report. PBC and PSC are characterized by inflammation and fibrosis of small intrahepatic (PBC) or larger intra- and/or extrahepatic (PSC) bile ducts. Bile duct damage leads to cholestasis and can progress to liver fibrosis and even cirrhosis. Various genetic, environmental and endogenous factors may contribute to the development of chronic cholestatic liver diseases, but the exact pathogenesis of PBC and PSC has not been clarified. Ursodeoxycholic acid (UDCA) is the standard treatment of PBC and is used also for other cholestatic conditions including PSC, and it exerts anticholestatic effects at adequate doses. Novel anticholestatic therapeutic options for patients not adequately responding to UDCA are under development or have, like obeticholic acid, already been proven to have efficacy when combined with UDCA in the treatment of PBC. The future role of immunomodulating/immunosuppressive drug regimens must be critically reviewed.


Advances in tenascin-C biology.
Tenascin-C is an extracellular matrix glycoprotein that is specifically and transiently expressed upon tissue injury. Upon tissue damage, tenascin-C plays a multitude of different roles that mediate both inflammatory and fibrotic processes to enable effective tissue repair. In the last decade, emerging evidence has demonstrated a vital role for tenascin-C in cardiac and arterial injury, tumor angiogenesis and metastasis, as well as in modulating stem cell behavior. Here we highlight the molecular mechanisms by which tenascin-C mediates these effects and discuss the implications of mis-regulated tenascin-C expression in driving disease pathology.


ErbB2 induces Notch1 activity and function in breast cancer cells.
The ErbB2 (Her2/neu epidermal growth receptor family) oncogene is overexpressed in 30% to 40% of human breast cancers. Cyclin D1 is the regulatory subunit of the holoenzyme that phosphorylates and inactivates the retinoblastoma (pRb) tumor suppressor and is an essential downstream target of ErbB2-induced tumor growth. Herein, we demonstrate that ErbB2 induces the activity of the Notch signaling pathway. ErbB2 induction of DNA synthesis, contact-independent growth, and mammosphere induction required Notch1. ErbB2-induced cyclin D1 and cyclin D1 expression was suficient to induce Notch1 activity, and conversely, genetic deletion of Notch1 in mammary epithelial cells using foxed Notch (Notch(fl/fl)) mice demonstrated that cyclin D1 is induced by Notch1. Genetic deletion of cyclin D1 or small interfering RNA (siRNA) to cyclin D1-reduced Notch1 activity and reintroduction of cyclin D1 into cyclin D1-deficient cells restored Notch1 activity through the inhibition of Numb, an endogenous inhibitor of Notch1 activity. Thus, cyclin D1 functions downstream as a genetic target of Notch1, amplifies Notch1 activity by repressing Numb, and identifies a novel pathway by which ErbB2 induces Notch1 activity via the induction of cyclin D1.


A Smartphone App for Improving Mental Health through Connecting with Urban Nature.
In an increasingly urbanised world where mental health is currently in crisis, interventions to increase human engagement and connection with the natural environment are one of the fastest growing, most widely accessible, and cost-effective ways of improving human wellbeing. This study aimed to provide an evaluation of a smartphone app-based wellbeing intervention. In a randomised controlled trial study design, the app prompted 582 adults, including a subgroup of adults classified by baseline scores on the Recovering Quality of Life scale as having a common mental health problem (n = 148), to notice the good things about urban nature (intervention condition) or built spaces (active control). There were statistically significant and sustained improvements in wellbeing at one-month follow-up. Importantly, in the noticing urban nature condition, compared to a built space control, improvements in quality of life reached statistical significance for all adults and clinical significance for those classified as having a mental health difficulty. This improvement in wellbeing was partly explained by significant increases in nature connectedness and positive affect. This study provides the first controlled experimental evidence that noticing the good things about urban nature has strong clinical potential as a wellbeing intervention and social prescription.


Identification of genes downstream of Pax6 in the mouse lens using cDNA microarrays.
Pax6 is a transcription factor that regulates the development of the visual, olfactory, and central nervous systems, pituitary, and pancreas. Pax6 is required for induction, growth, and maintenance of the lens; however, few direct Pax6 target genes are known. This study was designed to identify batteries of differentially expressed genes in three related systems: 8-week old Pax6 heterozygous lenses, 8-week old Pax6 heterozygous eyes, and transgenic lenses overexpressing PAX6(5a), using high throughput cDNA microarrays containing about 9700 genes. Initially, we obtained almost 400 differentially expressed genes in lenses from mice heterozygous for a Pax6 deletion, suggesting that Pax6 haploinsufficiency causes global changes in the lens transcriptome. Comparisons between the three sets of analyses revealed that paralemmin, molybdopterin synthase sulfurylase, Tel6 oncogene (ETV6), a cleavage-specific factor (Cpsf1) and tangerin A were abnormally expressed in all three experimental models. Semiquantitative reverse transcription (RT)-PCR analysis confirmed that all five of these genes were differentially expressed in Pax-6 heterozygous and Pax6(5a) transgenic lenses. Western blotting and immunohistochemistry demonstrated that paralemmin is found at high levels in the adult lens and confirmed its down-regulation in the Pax6(5a)-transgenic lenses. Collectively, our data provide insights into the genetic programs regulated by Pax6 in the lens.


European evidence-based recommendations for the diagnosis and treatment of childhood-onset lupus nephritis: the SHARE initiative.
Lupus nephritis (LN) occurs in 50%-60% of patients with childhood-onset systemic lupus erythematosus (cSLE), leading to significant morbidity. Timely recognition of renal involvement and appropriate treatment are essential to prevent renal damage. The Single Hub and Access point for paediatric Rheumatology in Europe (SHARE) initiative aimed to generate diagnostic and management regimens for children and adolescents with rheumatic diseases including cSLE. Here, we provide evidence-based recommendations for diagnosis and treatment of childhood LN. Recommendations were developed using the European League Against Rheumatism standard operating procedures. A European-wide expert committee including paediatric nephrology representation formulated recommendations using a nominal group technique. Six recommendations regarding diagnosis and 20 recommendations covering treatment choices and goals were accepted, including each class of LN, described in the International Society of Nephrology/Renal Pathology Society 2003 classification system. Treatment goal should be complete renal response. Treatment of class I LN should mainly be guided by other symptoms. Class II LN should be treated initially with low-dose prednisone, only adding a disease-modifying antirheumatic drug after 3 months of persistent proteinuria or prednisone dependency. Induction treatment of class III/IV LN should be mycophenolate mofetil (MMF) or intravenous cyclophosphamide combined with corticosteroids; maintenance treatment should be MMF or azathioprine for at least 3 years. In pure class V LN, MMF with low-dose prednisone can be used as induction and MMF as maintenance treatment. The SHARE recommendations for diagnosis and treatment of LN have been generated to support uniform and high-quality care for all children with SLE.


Factors associated with the onset and persistence of post-lumbar puncture headache.
This study assesses factors associated with the most common adverse event following lumbar puncture. To identify factors associated with the risk, onset, and persistence of post-dural puncture headache (PDPH). We performed univariate and multivariable analyses of 338 lumbar punctures in the Dominantly Inherited Alzheimer Network observational study using linear mixed models, adjusting for participant-level and family-level random effects. We directly evaluated associations of 3 post-lumbar puncture outcomes (immediate postprocedural headache, PDPH at 24-hour follow-up, and PDPH receiving a therapeutic blood patch) with participant age and sex, positioning, collection method, needle size, needle insertion site, and cerebrospinal fluid (CSF) volume collected. The incidence of adverse events included 73 immediate postprocedural headaches (21.6%), 59 PDPHs at 24-hour follow-up (17.5%), and 15 PDPHs receiving a therapeutic blood patch (4.4%). Greater volume of CSF collected was associated with increased risk of immediate postprocedural headache, largely owing to a nonlinear increase in risk on collection of volumes above 30 mL (odds ratio, 3.73 for >30 mL and 0.98 for <17 mL). In contrast, collection of higher volumes showed a protective effect in decreasing rates of PDPH at 24-hour follow-up and rates of PDPH receiving a therapeutic blood patch (odds ratio, 0.35 per 10 mL). Although differences in needle size did not reach statistical significance, no participant in the 24G needle group received a therapeutic blood patch compared to 8 of 253 for the larger 22G needles. Factors that acutely lower CSF pressure (eg, seated positioning or extracting very high volumes of CSF) may be associated with transient post-lumbar puncture headache, without increasing rates of persistent PDPH or therapeutic blood patch. Collection of up to 30 mL of CSF appears to be well tolerated and safe.


RhesusBase PopGateway: Genome-Wide Population Genetics Atlas in Rhesus Macaque.
Although population genetics studies have significantly accelerated the evolutionary and functional interrogations of genes and regulations, limited polymorphism data are available for rhesus macaque, the model animal closely related to human. Here, we report the first genome-wide effort to identify and visualize the population genetics profile in rhesus macaque. On the basis of the whole-genome sequencing of 31 independent macaque animals, we profiled a comprehensive polymorphism map with 46,146,548 sites. The allele frequency for each polymorphism site, the haplotype structure, as well as multiple population genetics parameters were then calculated on a genome-wide scale. We further developed a specific interface, the RhesusBase PopGateway, to facilitate the visualization of these annotations, and highlighted the applications of this highly integrative platform in clarifying the selection signatures of genes and regulations in the context of the primate evolution. Overall, the updated RhesusBase provides a comprehensive monkey population genetics framework for in-depth evolutionary studies of human biology.


Geographic variations in the whistles of bottlenose dolphins (Tursiops aduncus) along the east and west coasts of Australia.
Macrogeographic variations in the structure and repertoire of whistles from four geographically separated bottlenose dolphin (Tursiops aduncus) populations from the east and west coasts of Australia were investigated. Microgeographic variations were examined from two adjacent social groups from the same east coast population. Whistles were divided into five different tonal classes; sine, rise, down-sweep, flat and concave. The acoustic parameters of whistles; start, end, low, high and carrier frequencies and the number of inflection points and harmonics were measured and compared. These features were used to identify and define distinct whistle types. Differences in acoustic parameters of whistles between populations varied with the tonal class and the acoustic parameter being tested and were likely to be driven by localized environmental conditions. There were few differences in the acoustic parameters of whistles between social groups which was likely a result of vocal learning and mimicry from intermixing of individuals. There were small percentages of distinct whistle types that were shared between the both intermixing social groups and geographically separated populations, however, the majority of the whistle repertoires were unique. The development of the unique whistle repertoires may be primarily driven by the complex social organization of the bottlenose dolphin.


Patient and hospital characteristics associated with length of stay and hospital charges for pediatric sports-related injury hospitalizations in the United States, 2000-2003.
The objectives of this study were to describe the patient and hospital characteristics of pediatric sports injury hospitalization and to determine the hospital characteristics that are associated with length of stay and total hospital charges (per discharge). Children who were aged 5 to 18 years and had a diagnosis of a sports injury in the Nationwide Inpatient Sample between 2000 and 2003 were included. National estimates of pediatric sports injury hospitalization, including the average and median of length of stay and total hospital charges, were computed. The relationship of hospital characteristics to length of stay and total hospital charges were assessed using linear regression, adjusting for patient characteristics and sample weight. A total of 7979 pediatric sports injury hospitalizations among children who were aged 5 to 18 were identified during 4 years, approximately 10,000 per year, nationwide. More than half of the hospitalizations were attributed to fractures. The mean and median of length of stay for pediatric sports injuries was 2.4 and 1.1 days, respectively. When weighted, the estimated total hospital charges for sports injury hospitalizations among 5- to 18-year-olds were $485 million during 4 years, with a steady increase each year. Urban hospitals had 46.1% higher total hospital charges than rural hospitals. Hospitals in the western United States had significantly greater total hospital charges than those in other regions. The findings provide an empirical basis for future research on the magnitude of sports-related injuries that result in hospitalization among children. More research is needed to identify contributing factors that are associated with length of stay and total hospital charges for sports injury hospitalization. Intervention efforts also should be directed toward preventing severe sports injuries and to reducing the hospitalization and cost.