Cardiovascular dynamics and adrenergic responses of the rainbow trout in vivo.
Cardiac output and dorsal aorta, ventral aorta, caudal artery, caudal vein, and subintestinal vein pressures have been directly measured in intact unanaesthetized trout. Cardiac output (Q) averaged 36.7 ml/kg.min. The pressure drop across the systemic vascular resistance (Rs) was approximately twice that across the gill resistance (Rg), and a significant positive pressure persisted in the venous system. alpha-Adrenergic blockade revealed a considerable endogenous vasomotor tone resulting from latent adrenergic constriction of Rs. Intravenous adrenaline caused a pressor response throughout the circulatory system which has been analysed in detail with the aid of previous studies on isolated parts of the trout circulation. The complex and variable form of the pressor response reflected differential contributions from changes in Q, Rg, and Rs. Increases in Rs (alpha-receptor activation) were the principal cause of all pressor responses. Rg usually declined slightly due to passive dilation and/or beta-receptor stimulation, but occasionally increased due to alpha-receptor activation. The cardiac response reflected a varying balance between a direct beta-stimulatory effect of adrenaline on Q and an indirect passive inhibition of Q by the increase in peripheral resistance. Both effects were mediated through changes in stroke volume. Occasional tachycardia or more frequent reflex bradycardia were minor components of the cardiac response. The in vivo actions of other adrenergic agents have been similarly analysed.


A spatiotemporal analysis of enzymatic activities associated with carbon metabolism in wild-type and mutant embryos of Arabidopsis using in situ histochemistry.
Arabidopsis as a molecular genetic model offers many advantages for the study of seed development, but these do not extend to biochemical and enzymatic studies, which are often compromised by the limited amount of material available from the small developing embryos. A set of assays based on the coupling of an enzymatic reaction to the reduction of NAD, NADP or FAD, and subsequent reduction and precipitation of a tetrazolium salt, have been adapted to investigate 18 enzyme activities associated with carbon metabolism in developing Arabidopsis embryos. The use of organelle-specific marker enzymes demonstrates the utility of the method for detection of activities in mitochondria, plastids and peroxisomes as well as the cytosol. The temporal staining patterns obtained allow classification of the activities into three main categories based on whether they peak in the early, intermediate or late stages of maturation. An interesting switch from ATP to pyrophosphate consuming pathways occurs at the onset of the maturation phase, which involves key steps in primary carbon metabolism such as phosphofructokinase. This spatiotemporal characterization of carbon metabolism has also been applied to various mutants disrupted in embryo development including gnom (gn), acetyl-CoA carboxylase1 (acc1), schlepperless (slp), and wrinkled1 (wri1). The data obtained demonstrate that the extent to which carbon metabolism is affected in mutants is not necessarily correlated to the severity of the mutation considered. Through the advanced characterization of trehalose-6-P synthase1 (tps1) embryos, this approach finally provides new insight into the regulatory role played by trehalose metabolism in embryo development.


Non-classic congenital adrenal hyperplasia due to 21-hydroxylase deficiency revisited: an update with a special focus on adolescent and adult women.
Non-classic congenital hyperplasia (NCAH) due to 21-hydroxylase deficiency is a common autosomal recessive disorder characterized by androgen excess. We conducted a systematic review and critical assessment of the available evidence pertaining to the epidemiology, pathophysiology, diagnosis and management of NCAH. A meta-analysis of epidemiological data was also performed. Peer-reviewed studies evaluating NCAH published up to October 2016 were reviewed. Multiple databases were searched including MEDLINE, EMBASE, Cochrane, ERIC, EBSCO, dissertation abstracts, and current contents. The worldwide prevalence of NCAH amongst women presenting with signs and symptoms of androgen excess is 4.2% (95% confidence interval: 3.2-5.4%). The clinical consequences of NCAH expand from infancy, i.e. accelerated growth, to adolescence and adulthood, i.e. premature pubarche, cutaneous symptoms and oligo-ovulation in a polycystic ovary syndrome (PCOS)-like clinical picture. The diagnosis of NCAH relies on serum 17-hydroxyprogesterone (17-OHP) concentrations. A basal 17-OHP concentration ≥2 ng/ml (6 nmol/l) should be used for screening if more appropriate in-house cut-off values are not available. Definitive diagnosis requires a 17-OHP concentration ≥10 ng/ml (30 nmol/l), either basally or after cosyntropin-stimulation. Molecular genetic analysis of the CYP21A2 gene, which is responsible for 21-hydroxylase activity, may be used for confirmation purposes and should be offered to all patients with NCAH along with genetic counseling because these patients frequently carry alleles that may result in classic CAH, the more severe form of the disease, in their progeny. Treatment must be individualized. Glucocorticoid replacement therapy may benefit pediatric patients with accelerated growth or advanced bone age or adult women seeking fertility, whereas adequate control of menstrual irregularity, hirsutism and other cutaneous symptoms is best served by the use of oral contraceptive pills and/or anti-androgens. Some women may need ovulation induction or assisted reproductive technology to achieve pregnancy. Patients with NCAH have a higher risk of miscarriage and may benefit from glucocorticoid treatment during pregnancy. Evidence-based diagnostic and treatment strategies are essential for the proper management of women with NCAH, especially considering that these patients may need different therapeutic strategies at different stages during their follow-up and that appropriate genetic counseling may prevent the occurrence of CAH in their children.


Potential-induced enantioselective uptake of amino acid into molecularly imprinted overoxidized polypyrrole.
Overoxidized polypyrrole films templated with L-glutamate have been utilized for enantioselective detection of L- and D-glutamic acid. Various important fabrication factors controlling the performance of the overoxidized polypyrrole films have been investigated using fluorescence spectrometry in conjugation with the electrochemical quartz crystal microbalance technique. The measured fluorescence intensity was related to the concentration of glutamate taken up into the films. It was found that L-glutamate was inserted approximately 30 times higher into the film than D-glutamate. Several key parameters such as applied potential and pH of amino acid solution were varied to achieve the optimum sensor response. The sensor templated with L-glutamic acid also exhibited excellent selectivity over several other L- and D-amino acids. Higher enantioselectivity of overoxidized polypyrrole film than that for previously reported imprinted polymers can be attributed to the potential-induced uptake/release of targeted molecules.


Modeling the Justinianic Plague: Comparing hypothesized transmission routes.
The Justinianic Plague, the first part of the earliest of the three plague pandemics, has minimal historical documentation. Based on the limited primary sources, historians have argued both for and against the "maximalist narrative" of plague, i.e. that the Justinianic Plague had universally devastating effects throughout the Mediterranean region during the sixth century CE. Using primary sources of one of the pandemic's best documented outbreaks that took place in Constantinople during 542 CE, as well as modern findings on plague etiology and epidemiology, we developed a series of dynamic, compartmental models of disease to explore which, if any, transmission routes of plague are feasible. Using expected parameter values, we find that the bubonic and bubonic-pneumonic transmission routes exceed maximalist mortality estimates and are of shorter detectable duration than described by the primary sources. When accounting for parameter uncertainty, several of the bubonic plague model configurations yielded interquartile estimates consistent with the upper end of maximalist estimates of mortality; however, these models had shorter detectable outbreaks than suggested by the primary sources. The pneumonic transmission routes suggest that by itself, pneumonic plague would not cause significant mortality in the city. However, our global sensitivity analysis shows that predicted disease dynamics vary widely for all hypothesized transmission routes, suggesting that regardless of its effects in Constantinople, the Justinianic Plague would have likely had differential effects across urban areas around the Mediterranean. Our work highlights the uncertainty surrounding the details in the primary sources on the Justinianic Plague and calls into question the likelihood that the Justinianic Plague affected all localities in the same way.


Breast cancer subtype dictates DNA methylation and ALDH1A3-mediated expression of tumor suppressor RARRES1.
Breast cancer subtyping, based on the expression of hormone receptors and other genes, can determine patient prognosis and potential options for targeted therapy. Among breast cancer subtypes, tumors of basal-like and claudin-low subtypes are typically associated with worse patient outcomes, are primarily classified as triple-negative breast cancers (TNBC), and cannot be treated with existing hormone-receptor-targeted therapies. Understanding the molecular basis of these subtypes will lead to the development of more effective treatment options for TNBC. In this study, we focus on retinoic acid receptor responder 1 (RARRES1) as a paradigm to determine if breast cancer subtype dictates protein function and gene expression regulation. Patient tumor dataset analysis and gene expression studies of a 26 cell-line panel, representing the five breast cancer subtypes, demonstrate that RARRES1 expression is greatest in basal-like TNBCs. Cell proliferation and tumor growth assays reveal that RARRES1 is a tumor suppressor in TNBC. Furthermore, gene expression studies, Illumina HumanMethylation450 arrays, and chromatin immunoprecipitation demonstrate that expression of RARRES1 is retained in basal-like breast cancers due to hypomethylation of the promoter. Additionally, expression of the cancer stem cell marker, aldehyde dehydrogenase 1A3, which provides the required ligand (retinoic acid) for RARRES1 transcription, is also specific to the basal-like subtype. We functionally demonstrate that the combination of promoter methylation and retinoic acid signaling dictates expression of tumor suppressor RARRES1 in a subtype-specific manner. These findings provide a precedent for a therapeutically-inducible tumor suppressor and suggest novel avenues of therapeutic intervention for patients with basal-like breast cancer.


Obesity and other correlates of physical activity and sedentary behaviors among US high school students.
Understanding correlates of physical activity (PA) can help inform and improve programs that promote PA among youth. We analyzed data from the 2010 National Youth Physical Activity and Nutrition Study, a representative sample of US students in grades 9-12. Logistic regression was used to examine associations between PA correlates (obesity, physical education classes, sports team participation, attitude toward PA, adult support for PA, and environmental support for PA) and participation in daily PA (DPA), vigorous PA (VPA), muscle-strengthening activity (MSA), viewing television (TV), and using computers or video games (C/VG). A positive attitude toward PA and adult support for PA were both associated with increased PA and decreased sedentary behavior. However, among students who lived in neighborhoods that were not safe for PA, a positive attitude toward PA was not associated with increased DPA or decreased sedentary behavior and was less strongly associated with VPA and MSA. Efforts to increase PA among youth should promote a positive attitude toward PA among youth and encourage adult family members to support their efforts to be active. Policies that promote safe neighborhoods may work synergistically with a positive attitude toward PA to increase participation in PA and decrease sedentary behaviors.


The lack of maturation of Ebola virus-infected dendritic cells results from the cooperative effect of at least two viral domains.
Ebola virus (EBOV) infections are characterized by deficient T lymphocyte responses, T lymphocyte apoptosis, and lymphopenia in the absence of direct infection of T lymphocytes. In contrast, dendritic cells (DC) are infected but fail to mature appropriately, thereby impairing the T cell response. We investigated the contributions of EBOV proteins in modulating DC maturation by generating recombinant viruses expressing enhanced green fluorescent protein and carrying mutations affecting several potentially immunomodulating domains. They included envelope glycoprotein (GP) domains, as well as innate response antagonist domains (IRADs) previously identified in the VP24 and VP35 proteins. GP expressed by an unrelated vector, but not the wild-type EBOV, was found to strongly induce DC maturation, and infections with recombinant EBOV carrying mutations disabling GP functional domains did not restore DC maturation. In contrast, each of the viruses carrying mutations disabling any IRAD in VP35 induced a dramatic upregulation of DC maturation markers. This was dependent on infection, but not interaction with GP. Disabling of IRADs also resulted in up to a several hundredfold increase in secretion of cytokines and chemokines. Furthermore, these mutations induced formation of homotypic DC clusters, which represent close correlates of their maturation and presumably facilitate transfer of antigen from migratory DC to lymph node DC. Thus, an individual IRAD is insufficient to suppress DC maturation; rather, the suppression of DC maturation and the "immune paralysis" observed during EBOV infections results from a cooperative effect of two or more individual IRADs.


Photorealistic rendering of unidirectional free-space invisibility cloaks.
Carpet or ground-plane invisibility cloaks hide an object in reflection and inhibit transmission experiments by construction. This concept has significantly reduced the otherwise demanding material requirements and has hence enabled various experimental demonstrations. In contrast, free-space invisibility cloaks should work in both reflection and transmission. The fabrication of omnidirectional three-dimensional free-space cloaks still poses significant challenges. Recently, the idea of the carpet cloak has been carried over to experiments on unidirectional free-space invisibility cloaks that only work perfectly for one particular viewing direction and, depending on the design, also for one linear polarization of light only. Here, by using photorealistic ray tracing, we visualize the performance of four types of such unidirectional cloaks in three dimensions for different viewing directions and different polarizations of light, revealing virtues and limitations of these approaches in an intuitive manner.


Critical examination of evidence for the nutritional status of children in Papua New Guinea - a systematic review.
Undernutrition remains a significant cause of childhood illness, poor growth, development, and death in Papua New Guinea (PNG). Studies on child nutritional outcomes in PNG vary by design, measurement protocols and quality. We conducted a systematic review to assess the evidence for the prevalence of child undernutrition across different study populations, geographical locations and time periods. Six electronic databases and additional grey literature were searched for articles describing the nutritional status by wasting, stunting and underweight, of PNG children under five years of age, published between 1990 and April 2015. Prevalence data using different scales of measurement and reference populations were standardized according to WHO protocols. The search yielded 566 articles, of which, 31 studies met the inclusion criteria. The prevalence of child undernutrition varied from 1% to 76% for wasting (median 11%), 5% to 92% for stunting (median 51%), and 14% to 59% for underweight (median 32%). Wide variations exist according to the index used for measurement, the population characteristics and the geographical region in which they live. Prevalence estimates increase significantly when data using different scales of measurement and population references are standardized to the WHO protocols. Child undernutrition in PNG is regionally variable due to a complex interplay of poverty, disease, food-security, cultural, environmental and sociopolitical issues requiring a complex mix of solutions by governments, health systems and local communities. Area- specific surveys using multiple measures are necessary to inform local solutions for this important problem.


Association between low maternal low-density lipoprotein cholesterol levels in the second trimester and delivery of small for gestational age infants at term: a case-control study of the national center for child health and development birth cohort.
Maternal malnutrition during pregnancy can lead to a small for gestational age (SGA) status among infants. In the present study, we compared the lipid levels during the second trimester in women who delivered SGA infants at term with those in women who delivered appropriate for gestational age infants at term, to examine whether abnormalities in maternal lipid levels could be a risk factor for the delivery of an SGA infant. We enrolled pregnant women who gave birth at the National Center for Child Health and Development (NCCHD), their infants, and who volunteered in a Birth cohort study at the NCCHD. We analyzed a total of 843 blood samples obtained from the women during the second trimester. Moreover, we used multi-regression analysis to assess the relationship between low-density lipoprotein cholesterol (LDL-C) levels during the second trimester and the risk of delivering an SGA infant. The adjusted odds ratio (OR) for LDL-C levels was 0.99 (95% CI, 0.98-0.99). These results indicate that a low LDL-C level during the second trimester was associated with an increased risk of delivering an SGA infant at term. The finding would not only help predict future SGA infant birth, but can also help prevent SGA infant birth.


Varicella-zoster virus thymidine kinase gene and antiherpetic pyrimidine nucleoside analogues in a combined gene/chemotherapy treatment for cancer.
Ten pyrimidine nucleoside analogues, including (B)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) and closely related analogues, were evaluated for their cytostatic activity against human osteosarcoma cells transfected with the varicella-zoster virus (VZV) thymidine kinase (tk) (ATP:thymidine 5' phosphotransferase, EC 184.108.40.206) gene. (E)-5-(2-bromovinyl)-1-beta-D-arabinofuranosyluracil (BVaraU), (E)-5-(2-iodovinyl)-2'-deoxy-2'-fluoro-1-beta-D-arabinofuranosyluracil (IVFAU) and (E)-5-(2-bromovinyl)-2'-deoxy-4'-thiouridine (S-BVDU) were among the most potent inhibitors of VZVtk gene-transfected cell proliferation. They displayed an inhibitory activity at drug concentrations that were up to four orders of magnitude lower than those required to inhibit the corresponding nontransfected tumor cells. Inhibition of cellular DNA polymerase and/or incorporation of the drugs into cellular DNA may be a likely target for the cytostatic activity of the BVDU derivatives against the VZVtk gene-transfected tumor cells. These compounds were approximately 40- to 80-fold more potent cytostatic agents in VZVtk gene-transfected cells than the anti-VZV compound 6-methoxy-9-beta-D-arabinofuranosylpurine (araM), and at least five- to 50-fold more cytostatic than ganciclovir in HSV-1tk gene-transfected murine mammary carcinoma FM3A cells. In addition, the intrinsic resistance of BVaraU, IVFAU and S-BVDU to glycosidic bond cleavage by mammalian dThd phosphorylases makes them promising candidate compounds for the treatment of VZVtk gene-transfected tumors in vivo.


Cortical neuronal and glial pathology in TgTauP301L transgenic mice: neuronal degeneration, memory disturbance, and phenotypic variation.
Recapitulation of tau pathologies in an animal model has been a long-standing goal in neurodegenerative disease research. We generated transgenic (TgTauP301L) mice expressing a frontotemporal dementia with parkinsonism linked to chromosome 17 (FTPD-17) mutation within the longest form of tau (2N, 4R). TgTauP301L mice developed florid pathology including neuronal pretangles, numerous Gallyas-Braak-positive neurofibrillary tangles, and glial fibrillary tangles in the frontotemporal areas of the cerebrum, in the brainstem, and to a lesser extent in the spinal cord. These features were accompanied by gliosis, neuronal loss, and cerebral atrophy. Accumulated tau was hyperphosphorylated, conformationally changed, ubiquitinated, and sarkosyl-insoluble, with electron microscopy demonstrating wavy filaments. Aged TgTauP301L mice exhibited impairment in hippocampally dependent and independent behavioral paradigms, with impairments closely related to the presence of tau pathologies and levels of insoluble tau protein. We conclude that TgTauP301L mice recreate the substantial phenotypic variation and spectrum of pathologies seen in FTDP-17 patients. Identification of genetic and/or environmental factors modifying the tau phenotype in these mice may shed light on factors modulating human tauopathies. These transgenic mice may aid therapeutic development for FTDP-17 and other diseases featuring accumulations of four-repeat tau, such as Alzheimer's disease, corticobasal degeneration, and progressive supranuclear palsy.


Management of extensive external apical root resorption leading to root perforation.
Resorption is both a physiological and pathological process which results in loss of hard tissues of the tooth, i.e , cementum and dentine and the surrounding bone. External resorption is much more common than internal resorption and can occur when tooth is luxated or avulsed. If remained unchecked, resorption can eventually lead to loss of the tooth. Timely management of the affected tooth can slow down the resorptive lesion and increase the prognosis of the survival of the tooth. This case report describes the surgical management of extensive external root resorption leading to perforation of apical one-third of the root area which was managed through root canal along with periapical surgery and bone grafting. A 6-month follow-up showed arrest of the resorptive defect and progressive healing as evident on the cone-beam computed tomography.


Geochemical Evidence for the Control of Fire by Middle Palaeolithic Hominins.
The use of fire played an important role in the social and technological development of the genus Homo. Most archaeologists agree that this was a multi-stage process, beginning with the exploitation of natural fires and ending with the ability to create fire from scratch. Some have argued that in the Middle Palaeolithic (MP) hominin fire use was limited by the availability of fire in the landscape. Here, we present a record of the abundance of polycyclic aromatic hydrocarbons (PAHs), organic compounds that are produced during the combustion of organic material, from Lusakert Cave, a MP site in Armenia. We find no correlation between the abundance of light PAHs (3-4 rings), which are a major component of wildfire PAH emissions and are shown to disperse widely during fire events, and heavy PAHs (5-6 rings), which are a major component of particulate emissions of burned wood. Instead, we find heavy PAHs correlate with MP artifact density at the site. Given that hPAH abundance correlates with occupation intensity rather than lPAH abundance, we argue that MP hominins were able to control fire and utilize it regardless of the variability of fires in the environment. Together with other studies on MP fire use, these results suggest that the ability of hominins to manipulate fire independent of exploitation of wildfires was spatially variable in the MP and may have developed multiple times in the genus Homo.


How well do physicians use electronic information retrieval systems? A framework for investigation and systematic review.
Despite the proliferation of electronic information retrieval (IR) systems for physicians, their effectiveness has not been well assessed. The purpose of this review is to provide a conceptual framework and to apply the results of previous studies to this framework. All sources of medical informatics and information science literature, including MEDLINE, along with bibliographies of textbooks in these areas, were searched from 1966 to January 1998. All articles presenting either classifications of evaluation studies or their results, with an emphasis on those studying use by physicians. A framework for evaluation was developed, consisting of frequency of use, purpose of use, user satisfaction, searching utility, search failure, and outcomes. All studies were then assessed based on the framework. Due to the heterogeneity and simplistic study designs, no meta-analysis of studies could be done. General conclusions were drawn from data where appropriate. A total of 47 articles were found to include an evaluation component and were used to develop the framework. Of these, 21 articles met the inclusion criteria for 1 or more of the categories in the framework. Most use of IR systems by physicians still occurs with bibliographic rather than full-text databases. Overall use of IR systems occurs just 0.3 to 9 times per physician per month, whereas physicians have 2 unanswered questions for every 3 patients. Studies comparing IR systems with different searching features have not shown that advanced searching methods are significantly more effective than simple text word methods. Most searches retrieve only one fourth to one half of the relevant articles on a given topic and, once retrieved, little is known about how these articles are interpreted or applied. These studies imply that further research and development are needed to improve system utility and performance.


Measurement of plasma colloid osmotic pressure in normal thoroughbred neonatal foals.
A normal plasma colloid osmotic pressure (COP) interval was established for foals and compared to values for adult horses. Plasma samples were obtained from 38 Thoroughbred foals that had normal findings on postfoaling examination and 10 healthy Thoroughbred adult horses. Samples were analyzed using a commercially available colloid osmometer. Fifty samples were obtained from 38 foals. Twelve foals had 2 samples taken, 1 during the 1st 24 hours of life and the 2nd between 24 and 72 hours of life. For foals with 2 samples, only 1 randomly selected value was used in group analysis. Total plasma protein, albumin, and globulin concentrations were measured on all samples from foals. The mean measured plasma COP for foals was 18.8 +/- 1.9 mm Hg for the 38 samples analyzed. Measured plasma COP did not differ significantly over the time period examined for either the 12 paired samples (P = .13) or with regression analysis of the 38 samples (P = .13). Calculation of mean COP, based on previously published quadratic equations using total protein, albumin, and globulin concentrations, underestimated mean measured foal COP values except for when total protein measured by refractometer was used in the Landis-Pappenheimer equation. In conclusion, the plasma COP interval (95% CI: 15.0 mm Hg, 22.6 mm Hg) obtained for healthy foals in this study was found to be lower than that of healthy adult Thoroughbreds (20.6 +/- 0.7 mm Hg, P = .006).


New approaches for carbon nanotubes-based biosensors and their application to cell culture monitoring.
Amperometric biosensors are complex systems and they require a combination of technologies for their development. The aim of the present work is to propose a new approach in order to develop nanostructured biosensors for the real-time detection of multiple metabolites in cell culture flasks. The fabrication of five Au working electrodes onto silicon substrate is achieved with CMOS compatible microtechnology. Each working electrode presents an area of 0.25 mm², so structuration with carbon nanotubes and specific functionalization are carried out by using spotting technology, originally developed for microarrays and DNA printing. The electrodes are characterized by cyclic voltammetry and compared with commercially available screen-printed electrodes. Measurements are carried out under flow conditions, so a simple fluidic system is developed to guarantee a continuous flow next to the electrodes. The working electrodes are functionalized with different enzymes and calibrated for the real-time detection of glucose, lactate, and glutamate. Finally, some tests are performed on surnatant conditioned medium sampled from neuroblastoma cells (NG-108 cell line) to detect glucose and lactate concentration after 72 hours of cultivation. The developed biosensor for real-time and online detection of multiple metabolites shows very promising results towards circuits and systems for cell culture monitoring.


Nanostructuration strategies to enhance microelectrode array (MEA) performance for neuronal recording and stimulation.
Microelectrode arrays (MEAs) are widely used tools for recording and stimulating extracellular neuronal activity. Major limitations when decreasing electrode size in dense arrays are increased noise level and low charge injection capability. Nanostructuration of the electrode sites on MEAs presents an efficient way to overcome these problems by decreasing the impedance of the electrode/solution interface. Here, we review different techniques used to achieve this goal including template assisted electrodeposition for generating macro- and mesoporous films, immobilization of carbon nanotubes (CNTs) and deposition of conducting polymers onto microelectrodes. When tested during in vitro and in vivo measurements, nanostructured MEAs display improved sensitivity during recording of neuronal activity together with a higher efficiency in the stimulation process compared to conventional microelectrodes.


Linking the oceans to public health: current efforts and future directions.
We review the major linkages between the oceans and public health, focusing on exposures and potential health effects due to anthropogenic and natural factors including: harmful algal blooms, microbes, and chemical pollutants in the oceans; consumption of seafood; and flooding events. We summarize briefly the current state of knowledge about public health effects and their economic consequences; and we discuss priorities for future research.We find that:* There are numerous connections between the oceans, human activities, and human health that result in both positive and negative exposures and health effects (risks and benefits); and the study of these connections comprises a new interdisciplinary area, "oceans and human health."* The state of present knowledge about the linkages between oceans and public health varies. Some risks, such as the acute health effects caused by toxins associated with shellfish poisoning and red tide, are relatively well understood. Other risks, such as those posed by chronic exposure to many anthropogenic chemicals, pathogens, and naturally occurring toxins in coastal waters, are less well quantified. Even where there is a good understanding of the mechanism for health effects, good epidemiological data are often lacking. Solid data on economic and social consequences of these linkages are also lacking in most cases.* The design of management measures to address these risks must take into account the complexities of human response to warnings and other guidance, and the economic tradeoffs among different risks and benefits. Future research in oceans and human health to address public health risks associated with marine pathogens and toxins, and with marine dimensions of global change, should include epidemiological, behavioral, and economic components to ensure that resulting management measures incorporate effective economic and risk/benefit tradeoffs.


The story of the lost twins: decoding the genetic identities of the Kumhar and Kurcha populations from the Indian subcontinent.
The population structure of the Indian subcontinent is a tapestry of extraordinary diversity characterized by the amalgamation of autochthonous and immigrant ancestries and rigid enforcement of sociocultural stratification. Here we investigated the genetic origin and population history of the Kumhars, a group of people who inhabit large parts of northern India. We compared 27 previously published Kumhar SNP genotype data sampled from Uttar Pradesh in north India to various modern day and ancient populations. Various approaches such as Principal Component Analysis (PCA), Admixture, TreeMix concurred that Kumhars have high ASI ancestry, minimal Steppe component and high genomic proximity to the Kurchas, a small and relatively little-known population found ~ 2500 km away in Kerala, south India. Given the same, biogeographical mapping using Geographic Population Structure (GPS) assigned most Kumhar samples in areas neighboring to those where Kurchas are found in south India. We hypothesize that the significant genomic similarity between two apparently distinct modern-day Indian populations that inhabit well separated geographical areas with no known overlapping history or links, likely alludes to their common origin during or post the decline of the Indus Valley Civilization (estimated by ALDER). Thereafter, while they dispersed towards opposite ends of the Indian subcontinent, their genomic integrity and likeness remained preserved due to endogamous social practices. Our findings illuminate the genomic history of two Indian populations, allowing a glimpse into one or few of numerous of human migrations that likely occurred across the Indian subcontinent and contributed to shape its varied and vibrant evolutionary past.


Excitotoxicity triggered by Neurobasal culture medium.
Neurobasal defined culture medium has been optimized for survival of rat embryonic hippocampal neurons and is now widely used for many types of primary neuronal cell culture. Therefore, we were surprised that routine medium exchange with serum- and supplement-free Neurobasal killed as many as 50% of postnatal hippocampal neurons after a 4 h exposure at day in vitro 12-15. Minimal Essential Medium (MEM), in contrast, produced no significant toxicity. Detectable Neurobasal-induced neuronal death occurred with as little as 5 min exposure, measured 24 h later. D-2-Amino-5-phosphonovalerate (D-APV) completely prevented Neurobasal toxicity, implicating direct or indirect N-methyl-D-aspartate (NMDA) receptor-mediated neuronal excitotoxicity. Whole-cell recordings revealed that Neurobasal but not MEM directly activated D-APV-sensitive currents similar in amplitude to those gated by 1 µM glutamate. We hypothesized that L-cysteine likely mediates the excitotoxic effects of Neurobasal incubation. Although the original published formulation of Neurobasal contained only 10 µM L-cysteine, commercial recipes contain 260 µM, a concentration in the range reported to activate NMDA receptors. Consistent with our hypothesis, 260 µM L-cysteine in bicarbonate-buffered saline gated NMDA receptor currents and produced toxicity equivalent to Neurobasal. Although NMDA receptor-mediated depolarization and Ca²⁺ influx may support survival of young neurons, NMDA receptor agonist effects on development and survival should be considered when employing Neurobasal culture medium.


Transcutaneous electrical nerve stimulation and acupuncture-like transcutaneous electrical nerve stimulation for chronic low back pain.
Transcutaneous electrical nerve stimulation (TENS), originally based on the gate-control theory of pain, is widely used for the treatment of chronic low back pain. Despite its wide use and theoretical rationale, there appears at first glance little scientific evidence to support its use. This Cochrane review examines the available evidence on TENS for the treatment of chronic back pain through an exhaustive search of the literature. Transcutaneous electrical nerve stimulation (TENS) and acupuncture-like transcutaneous electrical nerve stimulation (ALTENS) for chronic low back pain management have experienced a tremendous growth over the past 25 years. The objective of this review was to assess the effects of TENS and ALTENS for reducing pain and improving function in patients with chronic back pain. We searched MEDLINE up to November 1997, EMBASE from 1985 to September 1995, Amed and Ciscom to January 1995, reference lists of the retrieved articles, proceedings of conferences and contacted investigators in the field. Randomised trials comparing TENS or ALTENS therapy to placebo in patients with chronic low back pain. Two reviewers independently assessed trial quality and extracted data on pain reduction, range of movement, functional and work status. Six trials were included. The trials included 288 participants with an average age range of 45 to 50 years and approximately equal numbers of women and men. The overall odds ratio for improvement in pain for each comparison was: TENS/ALTENS versus placebo 2.11 (95% confidence interval 1.32 to 3. 38), ALTENS versus placebo 7.22 (95% confidence interval 2.60 to 20.01) and TENS versus placebo 1.52 (95% confidence interval 0.90 to 2.58). The odds ration for improvement in range of motion on ALTENS versus placebo was 6.61 (95% confidence interval 2.36 to 18.55). There is evidence from the limited data available that TENS/ALTENS reduces pain and improves range of motion in chronic back pain patients, at least in the short term. A large trial of ALTENS and TENS is needed to confirm these findings.


High-performance tellurium-free thermoelectrics: all-scale hierarchical structuring of p-type PbSe-MSe systems (M = Ca, Sr, Ba).
We present a systematic study of the characterization and thermoelectric properties of nanostructured Na-doped PbSe embedded with 1-4% MSe (M = Ca, Sr, Ba) phases as endotaxial inclusions. The samples were powder-processed by the spark plasma sintering technique, which introduces mesoscale-structured grains. The hierarchical architectures on the atomic scale (Na and M solid solution), nanoscale (MSe nanoprecipitates), and mesoscale (grains) were confirmed by transmission electron microscopy. These structures produce a great reduction in the lattice thermal conductivity relative to pristine PbSe without appreciably affecting the power factor. The lattice thermal conductivity can be reduced by up to ∼29% when the second phase is added. The highest ZT value achieved was ∼1.3 at 923 K for both 2% SrSe-and 3% BaSe-containing samples, while the sample containing 4% CaSe showed a ZT value of ∼1.2 at 923 K. The optimal samples have hole carrier concentration of 1-2 × 10(20) cm(-3). We attribute the high ZT values to the combination of broad-based phonon scattering on multiple length scales and favorable charge transport through coherent interfaces between the PbSe matrix and MSe.


Pulsed laser excited photothermal lens spectrometry of CdSxSe(1-x) doped silica glasses.
Experimental results for photothermal lens measurements are compared to finite elemental analysis models for commercial colored glass filters. Finite elemental analysis software is used to model the photothermal effect by simulating the coupling of heat both within the sample and out to the surroundings. Modeling shows that heat transfer between the glass surface and the air coupling fluid has a significant effect on the predicted time-dependent photothermal lens signals. For comparison with experimental signals, a simple equation based on the finite element analysis result is proposed for accounting for the variance of experimental data where this type of heat coupling situation occurs. The colored glass filters are found to have positive thermo-optical coefficients. The net positive dn/dT of CdSxSe(1-x) doped glass filters is considered to be the consequence of counteracting factors: optical nonlinearity, stress-induced birefringence, and the structural network of glass. Finite element analysis modeling results are also used to correlate experimental measurements of different sample geometries. In particular, the glass samples are compared to ethanol solutions of iron (II) dicylopentadiene in a sample cuvette even though heat transfer is different for these two samples.


Challenges in malaria control in sub-Saharan Africa: the vaccine perspective.
Malaria is a life-threatening disease of public health importance, especially in sub-Saharan Africa. It is estimated that about 500 million cases of malaria occur annually and among these 1 million die annually. Children below five years and pregnant women are the most vulnerable groups. Several malaria control measures have been applied such as environmental improvements, use of insecticide impregnated nets, residual indoor spraying, early case detection and treatment with effective antimalarial drugs. However, the adaptation of vector and parasite has so far limited the effect of these interventions. The emergence of resistance against drugs and insecticides requires in response a steady stream of new interventions. Up to the beginning of this millennium, most sub-Saharan African countries have been using chloroquine (CQ) as the first-line antimalarial drug, which had to be replaced with sulphadoxine-pyrimethamine (SP) after resistant parasites had rendered CQ ineffective. Currently the first line treatment of malaria consists of combination therapy which includes an artemisinin derivative. The current approach appears robust but history has taught us to be alert and to expect resistance to emerge. There is a pressing need to develop and deploy complimentary strategies. Adding a protective vaccine to the existing control tools for malaria holds great promise yet no malaria vaccine has ever been licensed despite a large number of attempts. The complexity of malaria parasites and the ability of the parasite to suppress and evade immune responses are formidable challenges. Fortunately, there are several promising antimalarial vaccine candidates in the development pipeline. The most promising vaccine candidate is RTSS which is currently tested in various countries in sub-Saharan Africa, including two sites in Tanzania. There is a hope that malaria vaccines could be developed and deployed in malaria endemic communities. This article highlights the challenges of developing and deploying malaria vaccines.


Participation of GATA-3 in regulation of bone healing through transcriptional upregulation of bcl-xL expression.
We have previously demonstrated the expression of GATA-DNA-binding protein (GATA)-3, a transcription factor, in osteoblasts and have verified its function in transducing cell survival signaling. This translational study was further designed to evaluate the roles of GATA-3 in regulating bone healing and to explore its possible mechanisms. A metaphyseal bone defect was created in the left femurs of male ICR mice. Analysis by micro-computed topography showed that the bone volume, trabecular bone number and trabecular thickness were augmented and that the trabecular pattern factor decreased. Interestingly, immunohistological analyses showed specific expression of GATA-3 in the defect area. In addition, colocalized expression of GATA-3 and alkaline phosphatase was observed at the wound site. As the fracture healed, the amounts of phosphorylated and non-phosphorylated GATA-3 concurrently increased. Separately, GATA-3 mRNA was induced during bone healing, and, levels of Runx2 mRNA and protein were also increased. The results of confocal microscopy and co-immunoprecipitation showed an association between nuclear GATA-3 and Runx2 in the area of insult. In parallel with fracture healing, Bcl-XL mRNA was significantly triggered. A bioinformatic search revealed the existence of a GATA-3-specific DNA-binding element in the promoter region of the bcl-xL gene. Analysis by chromatin immunoprecipitation assays further demonstrated transactivation activity by which GATA-3 regulated bcl-xL gene expression. Therefore, this study shows that GATA-3 participates in the healing of bone fractures via regulating bcl-xL gene expression, owing to its association with Runx2. In the clinic, GATA-3 may be used as a biomarker for diagnoses/prognoses or as a therapeutic target for bone diseases, such as bone fractures.


A randomized feasibility trial comparing four antimalarial drug regimens to induce Plasmodium falciparum gametocytemia in the controlled human malaria infection model.
Malaria elimination strategies require a thorough understanding of parasite transmission from human to mosquito. A clinical model to induce gametocytes to understand their dynamics and evaluate transmission-blocking interventions (TBI) is currently unavailable. Here, we explore the use of the well-established Controlled Human Malaria Infection model (CHMI) to induce gametocyte carriage with different antimalarial drug regimens. In a single centre, open-label randomised trial, healthy malaria-naive participants (aged 18–35 years) were infected with Plasmodium falciparum by bites of infected Anopheles mosquitoes. Participants were randomly allocated to four different treatment arms (n = 4 per arm) comprising low-dose (LD) piperaquine (PIP) or sulfadoxine-pyrimethamine (SP), followed by a curative regimen upon recrudescence. Male and female gametocyte densities were determined by molecular assays. Mature gametocytes were observed in all participants (16/16, 100%). Gametocytes appeared 8.5–12 days after the first detection of asexual parasites. Peak gametocyte densities and gametocyte burden was highest in the LD-PIP/SP arm, and associated with the preceding asexual parasite biomass (p=0.026). Male gametocytes had a mean estimated circulation time of 2.7 days (95% CI 1.5–3.9) compared to 5.1 days (95% CI 4.1–6.1) for female gametocytes. Exploratory mosquito feeding assays showed successful sporadic mosquito infections. There were no serious adverse events or significant differences in the occurrence and severity of adverse events between study arms (p=0.49 and p=0.28). The early appearance of gametocytes indicates gametocyte commitment during the first wave of asexual parasites emerging from the liver. Treatment by LD-PIP followed by a curative SP regimen, results in the highest gametocyte densities and the largest number of gametocyte-positive days. This model can be used to evaluate the effect of drugs and vaccines on gametocyte dynamics, and lays the foundation for fulfilling the critical unmet need to evaluate transmission-blocking interventions against falciparum malaria for downstream selection and clinical development. Funded by PATH Malaria Vaccine Initiative (MVI). NCT02836002.


Blood pressure regulation in pheochromocytoma.
Two sets of studies were performed in 13 patients with proved adrenal pheochromocytoma to test the hypothesis that the sympathetic nervous system (SNS) is active and might contribute to the hypertensive state. Similar studies were performed in 15 additional patients considered to have essential hypertension. In the first set, 13 patients with pheochromocytoma were subjected to head-up tilt to assess the activity of the SNS. This maneuver decreased diastolic blood pressure in only two; heart rate increased appropriately in all except one. Changes in plasma norepinephrine (NE) were variable and did not correlate with changes in blood pressure (BP) and heart rate (HR). In the second set, 10 patients with pheochromocytoma were given a single oral dose of clonidine (0.3 mg) to evaluate what role, if any, the SNS might contribute to the hypertensive state. Fifteen patients with essential hypertension were studied similarly for comparison. Clonidine produced significant decreases in BP and HR but left plasma renin activity unchanged in both groups. In essential hypertension, the cardiovascular responses were accompanied by significant reductions in plasma NE. By contrast, plasma NE was unchanged in patients with pheochromocytoma, despite similar reductions in BP and HR. These results suggest that the sympathetic reflexes are intact in pheochromocytoma, and that much of the hypertension associated with these tumors may be related to increased sympathetic activity.


A Preliminary Study on the Mechanical Reliability and Regeneration Capability of Artificial Bone Grafts in Oncologic Cases, With and Without Osteosynthesis.
Several bone grafts are available for clinical use, each with their own peculiar biological and mechanical properties. A new bone graft was obtained by combining mineral structures from natural bovine bones with bioresorbable polymers and cellular nutrients. The study aims to evaluate the clinical, biological and structural properties of this bone graft and its reliability in orthopedic oncology. 23 adult patients (age range 18-85 years) were treated between October 2016 and December 2018; the oncologicdiagnoses were heterogeneous. After surgical curettage and bone grafting, a clinical-radiological follow up was conducted. Radiographs were used to evaluate graft integration according to the usual bone healing and oncologic follow up. Local complications (infection, local recurrence, wound dehiscence, fracture or early reabsorption) were evaluated. The mean followup was of 18.34 ± 4.83 months. No fracture or infection occurred. One case of patellar Giant Cell Tumor (GCT) and one of proximal tibia low-grade chondrosarcoma recurred after about one year. Two wound dehiscences occurred (one required a local flap). Follow-up X-rays showed good to excellent graft integration in most patients (20 out of 21). The investigated graft has a mechanical and structural function that can allow early weight-bearing and avoid a preventive bone fixation (only needed in four patients in this series). The graft blocks are different for shapes and dimensions, but they can be customized by the producer or sawcut by the surgeon in the operating theatre to fit the residual bone cavity. The complication rate was low, and a rapid integration was observed with no inflammatory reaction in the surrounding tissues. Further studies are mandatory to confirm these promising results.


Modeling Asymmetric Cell Division in Caulobacter crescentus Using a Boolean Logic Approach.
Caulobacter crescentus is a model organism for the study of asymmetric division and cell type differentiation, as its cell division cycle generates a pair of daughter cells that differ from one another in their morphology and behavior. One of these cells (called stalked) develops a structure that allows it to attach to solid surfaces and is the only one capable of dividing, while the other (called swarmer) develops a flagellum that allows it to move in liquid media and divides only after differentiating into a stalked cell type. Although many genes, proteins, and other molecules involved in the asymmetric division exhibited by C. crescentus have been discovered and characterized for several decades, it remains as a challenging task to understand how cell properties arise from the high number of interactions between these molecular components. This chapter describes a modeling approach based on the Boolean logic framework that provides a means for the integration of knowledge and study of the emergence of asymmetric division. The text illustrates how the simulation of simple logic models gives valuable insight into the dynamic behavior of the regulatory and signaling networks driving the emergence of the phenotypes exhibited by C. crescentus. These models provide useful tools for the characterization and analysis of other complex biological networks.


Structural and Functional Characterization of the PaaI Thioesterase from Streptococcus pneumoniae Reveals a Dual Specificity for Phenylacetyl-CoA and Medium-chain Fatty Acyl-CoAs and a Novel CoA-induced Fit Mechanism.
PaaI thioesterases are members of the TE13 thioesterase family that catalyze the hydrolysis of thioester bonds between coenzyme A and phenylacetyl-CoA. In this study we characterize the PaaI thioesterase from Streptococcus pneumoniae (SpPaaI), including structural analysis based on crystal diffraction data to 1.8-Å resolution, to reveal two double hotdog domains arranged in a back to back configuration. Consistent with the crystallography data, both size exclusion chromatography and small angle x-ray scattering data support a tetrameric arrangement of thioesterase domains in solution. Assessment of SpPaaI activity against a range of acyl-CoA substrates showed activity for both phenylacetyl-CoA and medium-chain fatty-acyl CoA substrates. Mutagenesis of putative active site residues reveals Asn(37), Asp(52), and Thr(68) are important for catalysis, and size exclusion chromatography analysis and x-ray crystallography confirm that these mutants retain the same tertiary and quaternary structures, establishing that the reduced activity is not a result of structural perturbations. Interestingly, the structure of SpPaaI in the presence of CoA provides a structural basis for the observed substrate specificity, accommodating a 10-carbon fatty acid chain, and a large conformational change of up to 38 Å in the N terminus, and a loop region involving Tyr(38)-Tyr(39). This is the first time PaaI thioesterases have displayed a dual specificity for medium-chain acyl-CoAs substrates and phenylacetyl-CoA substrates, and we provide a structural basis for this specificity, highlighting a novel induced fit mechanism that is likely to be conserved within members of this enzyme family.


A new technique for diagnosis of acetabular cup loosening using computed tomography: preliminary experience in 10 patients.
Considerable migration of the acetabular cup is required for diagnosis of loosening by conventional radiography. We have developed a new clinically applicable method for assessment of cup loosening using computed tomography (CT). 10 patients scheduled for revision hip replacement due to suspected wear or loosening were scanned twice with CT under torsion loading of the prosthesis. Two independent examiners assessed each patient with respect to motion of the acetabular cup relative to the pelvis using CT volume registration. The CT measurements were compared to findings at revision surgery. The method was applicable in 8 of the 10 patients. 1 patient had a severe tremor. In 1 patient, surgery revealed that the hip was ankylotic due to massive ectopic bone formation. This left 8 patients that could be evaluated. 4 cups were loose at surgery, and 3 of these cups could be seen to be mobile by CT. 4 cups that were stable on revision were accurately diagnosed as not being mobile by CT. Movements of less than 1 millimeter between bone and prosthesis could not be distinguished from errors in CT acquisition and volume registration. There was good agreement between the two observers. Movement of loose acetabular cups during torsion loading could be detected using CT volume registration. It was sensitive to cup movement in 3 out of 4 cases of loose cups. The method was specific and yielded no false positive results.


Angiopoietin-like protein 8 in early pregnancy improves the prediction of gestational diabetes.
Screening high-risk individuals for gestational diabetes mellitus (GDM) in early pregnancy conventionally relies on established maternal risk factors; however, the sensitivity and specificity of these factors are not satisfactory. The present study aimed to determine whether the concentration of angiopoietin-like protein 8 (ANGPTL8), either alone or combined with other risk factors in early pregnancy, could be used to predict subsequent GDM. From August 2015 to January 2016, 474 women receiving prenatal care at around 12-16 weeks of gestation were recruited into the study. ANGPTL8 levels were measured at the first prenatal visit. All the participants received a 75 g OGTT during weeks 24-28 of gestation. ANGPTL8 levels in early pregnancy were considerably higher in women who developed GDM than those who maintained normal glucose tolerance (2822 ± 938 vs 2120 ± 1118 pg/ml, respectively; p < 0.0001). Multivariable logistic regression revealed that ANGPTL8 levels were significantly associated with risk of GDM independent of conventional risk factors. In addition, women in the highest quartile of ANGPTL8 concentration had an 8.75-fold higher risk of developing GDM compared with women in the lowest quartile (OR8.75, 95%CI 2.43, 31.58). More importantly, incorporating ANGPTL8 into the conventional prediction model significantly increased the AUC for prediction of GDM (0.772vs 0.725; p = 0.019). Our study suggests that ANGPTL8 levels in early pregnancy are significantly and independently associated with risk of GDM at 24-28 weeks of gestation. Combining ANGPTL8 levels with conventional risk factors could thus improve the prediction of GDM.


A novel non-protein-coding infection-specific gene family is clustered throughout the genome of Phytophthora infestans.
Phytophthora infestans is the cause of late blight, a devastating and re-emerging disease of potato. Significant advances have been made in understanding the biology of P. infestans, and in the development of molecular tools to study this oomycete. Nevertheless, little is known about the molecular bases of the establishment or development of disease in this hemibiotrophic pathogen. Suppression subtractive hybridization (SSH) was used to generate cDNA enriched for sequences upregulated during potato infection. To identify pathogen-derived cDNAs, and eliminate host sequences from further study, SSH cDNA was hybridized to a P. infestans bacterial artificial chromosome library. A new gene family was identified called Pinci1, comprising more than 400 members arranged in clusters of up to nine copies throughout the P. infestans draft genome sequence. Real-time RT-PCR was used to quantify the expression of five classes of transcript within the family, relative to the constitutively expressed PiactA gene, and it revealed them to be significantly upregulated from 12 to 33 h post-inoculation, a period defining the biotrophic phase of infection. Computational analysis of sequences suggested that transcripts were non-protein coding, and this was confirmed by transient expression of FLAG-tagged ORFs in P. infestans.


Cationic Microbubbles for Non-Selective Binding of Cavitation Nuclei to Bacterial Biofilms.
The presence of multi-drug resistant biofilms in chronic, persistent infections is a major barrier to successful clinical outcomes of therapy. The production of an extracellular matrix is a characteristic of the biofilm phenotype, intrinsically linked to antimicrobial tolerance. The heterogeneity of the extracellular matrix makes it highly dynamic, with substantial differences in composition between biofilms, even in the same species. This variability poses a major challenge in targeting drug delivery systems to biofilms, as there are few elements both suitably conserved and widely expressed across multiple species. However, the presence of extracellular DNA within the extracellular matrix is ubiquitous across species, which alongside bacterial cell components, gives the biofilm its net negative charge. This research aims to develop a means of targeting biofilms to enhance drug delivery by developing a cationic gas-filled microbubble that non-selectively targets the negatively charged biofilm. Cationic and uncharged microbubbles loaded with different gases were formulated and tested to determine their stability, ability to bind to negatively charged artificial substrates, binding strength, and, subsequently, their ability to adhere to biofilms. It was shown that compared to their uncharged counterparts, cationic microbubbles facilitated a significant increase in the number of microbubbles that could both bind and sustain their interaction with biofilms. This work is the first to demonstrate the utility of charged microbubbles for the non-selective targeting of bacterial biofilms, which could be used to significantly enhance stimuli-mediated drug delivery to the bacterial biofilm.


First Report of Daylily Rust in the United States.
In August 2000, rust symptoms were observed on the leaves of daylily plants (Hemerocallis sp. cv. Pardon Me) at a nursery in Dearing, GA. Based on urediniospore characters, the pathogen was tentatively identified as Puccinia hemerocallidis Thuem. Urediniospores were globose to ellipsoid and measured 19 to 30 × 17 to 22 μm (average size of 22 × 19 μm), corresponding to the previously reported description from Japan (1). Teliospores were absent from the sample but were found on daylily plants (cv. Star Struck) with symptoms similar to cv. Pardon Me from the same nursery in Dearing beginning in October 2000. However, the teliospores differed from those in the published description in that many one-celled teliospores (i.e., mesospores), measuring 32 to 43 × 14 to 19 μm (average size of 38 × 16 μm), were produced in addition to two-celled teliospores, which measured 41 to 53 × 16 to 21 μm (average size of 46 × 18 μm). Similar mesospores were present in a slide from an isotype specimen of P. hemerocallidis (US 72719) housed in the U.S. National Fungus Collection (Beltsville, MD). Daylily plants (cv. Pardon Me) were reinoculated with urediniospores by shaking infected plants over uninfected plants and exposing plants to 100% relative humidity for 24 h. Initial symptoms of small, discrete, yellow spots and streaks on the upper surfaces of leaves developed within 3 to 7 days, and uredia with urediniospores were evident at 7 to 14 days after inoculation. Daylily rust is native to Asia and may have been introduced into Georgia on plant materials sent from Central America. The original source of daylily rust is unclear because Central American producers also purchase and import plants from the United States for propagation and then sell divisions back to U.S. growers. Daylily rust is a disease of major concern for both daylily producers and gardeners. References: (1) N. Hiratsuka, et al. The Rust Flora of Japan. Tsukuba Shuppankai, Ibaraki, Japan, 1992.


Maternal mortality in the rural Gambia, a qualitative study on access to emergency obstetric care.
BACKGROUND: Maternal mortality is the vital indicator with the greatest disparity between developed and developing countries. The challenging nature of measuring maternal mortality has made it necessary to perform an action-oriented means of gathering information on where, how and why deaths are occurring; what kinds of action are needed and have been taken. A maternal death review is an in-depth investigation of the causes and circumstances surrounding maternal deaths. The objectives of the present study were to describe the socio-cultural and health service factors associated with maternal deaths in rural Gambia. METHODS: We reviewed the cases of 42 maternal deaths of women who actually tried to reach or have reached health care services. A verbal autopsy technique was applied for 32 of the cases. Key people who had witnessed any stage during the process leading to death were interviewed. Health care staff who participated in the provision of care to the deceased was also interviewed. All interviews were tape recorded and analyzed by using a grounded theory approach. The standard WHO definition of maternal deaths was used. RESULTS: The length of time in delay within each phase of the model was estimated from the moment the woman, her family or health care providers realized that there was a complication until the decision to seeking or implementing care was made. The following items evolved as important: underestimation of the severity of the complication, bad experience with the health care system, delay in reaching an appropriate medical facility, lack of transportation, prolonged transportation, seeking care at more than one medical facility and delay in receiving prompt and appropriate care after reaching the hospital. CONCLUSION: Women do seek access to care for obstetric emergencies, but because of a variety of problems encountered, appropriate care is often delayed. Disorganized health care with lack of prompt response to emergencies is a major factor contributing to a continued high mortality rate.


Intravenous administration of bone morphogenetic protein-7 after ischemia improves motor function in stroke rats.
We and others have previously reported that bone morphogenetic protein-7 (BMP-7), given before middle cerebral artery occlusion (MCAO), reduces ischemic injury in brain. Recent studies have indicated that receptors for BMP are upregulated after brain ischemia. It is possible that this upregulation may facilitate endogenous neurorepair in the ischemic brain. The purpose of this study was to determine the neuroregenerative effects of BMP-7 given parenterally after ischemia/reperfusion injury. Adult Sprague-Dawley rats were anesthetized with chloral hydrate. The middle cerebral artery was transiently occluded by a filament inserted through the right internal carotid artery. The filament was removed after 60-minute ischemia to allow reperfusion. Some animals were killed 24 hours after MCAO to examine BMP-7 mRNA expression. Other animals received a single dose of intravenous BMP-7 or vehicle at 24 hours after MCAO and were used for subsequent behavioral studies and BMP-7 immunostaining. BMP-7 mRNA was upregulated 24 hours after MCAO in untreated animals. BMP-7 immunoreactivity was dose-dependently increased on the ischemic side of the hippocampus/dentate on day 6 after MCAO in animals receiving intravenous injection of BMP-7. Animals receiving BMP-7 also showed a decrease in body asymmetry from day 7 to day 14 and an increase in locomotor activity on day 14 after MCAO. Our data indicate that BMP-7, given parenterally after stroke, can pass through the blood-brain barrier on the ischemic side and induce behavioral recovery in stroke animals at longer testing times.


Cutaneous lymphoproliferation and lymphomas in interleukin 7 transgenic mice.
To investigate the role of interleukin 7 (IL-7) in the development of the lymphoid system, we have generated two lines of transgenic mice carrying an IL-7 cDNA fused to an immunoglobulin heavy chain promoter and enhancer. This transgene is expressed in the bone marrow, lymph nodes, spleen, thymus, and skin provoking a perturbation of T cell development characterized by a marked reduction of CD4+ CD8+ (double-positive) thymocytes. Quite unexpectedly, however, both lines also develop a progressive cutaneous disorder involving a dermal lymphoid infiltrate that results in progressive alopecia, hyperkeratosis, and exfoliation. Although the infiltrate is primarily composed of T lineage cells, its development is not impeded in the athymic nu/nu background. Furthermore, the phenotype can be transmitted horizontally by transplanting lymphoid tissues or skin to syngeneic wild-type mice. Thus, the phenotype is conveyed by skin-homing, mobile cells (presumably the infiltrating lymphocytes) in a cell-autonomous fashion. In addition to the skin phenotype, this transgene also provokes the development of a lymphoproliferative disorder that induces B and T cell lymphomas within the first 4 mo of life. These findings suggest potential physiologic actions of IL-7 in T cell development and in cutaneous immunity. They also demonstrate that IL-7 can act as an oncogene in the living organism.


[In Alzheimer's disease, the clinical expression of behavioral and psychological signs and symptoms is early and specific of neuropathological stages].
The early diagnosis of Alzheimer's disease is a new challenge. This study concerns 50 patients, 34 females (68 %) and 16 males (32 %) with Alzheimer (AD), according to NINCDS-ADRDA diagnostic criteria. To systematically evaluate in all patients behavioral and psychological signs and symptoms of dementia (BPSSD), according to the stage of AD, with the patients of our population separated into two MMS groups. The first group was composed of patients with an MMS score from 10 to 20 (eight males and 19 females). Patients of the second group had an MMS score between 21 and 28 (eight males and 19 females). The Neuro-Psychiatric Inventory (NPI) was used to collect information on the presence of BPSSD in AD patients. NPI scores were correlated to the cognitive part of the Alzheimer's Disease Assessment Scale (ADAS-Cog) that permits evaluation of the severity of cognitive impairment in AD patients. Before starting the study, all patients gave their informed consent to participate in the study of BPSSD in AD. Statistical treatment of data was performed using STATVIEW. Our study demonstrates that BPSSD are present not only in early but also in moderate stages of AD. As cognitive impairment, BPSSD are an integrate part of the clinical picture. With a frequency of 74 % for the whole population, "anxiety" represented the more predominant BPSSD for all our patients at all stages of AD. At the very early stages of AD, BPSSD appeared to precede cognitive disorders. The symptomatic association of "depression", "agitation", and "irritability of mood" may remain in a steady state for a few months before the appearance of verbal episodic memory impairment, which is characteristic of hippocampus involvement. "Irritability" seems to specifically characterise the initial phase of AD. On the other hand, two BPSSD are characteristic of the late stages of AD: "sleep disorder" and "hallucinations".


Emerging Technologies to Extract Fucoxanthin from Undaria pinnatifida: Microwave vs. Ultrasound Assisted Extractions.
Macroalgae are an extensive resource for the obtention of bioactive compounds, mainly phenolic compounds, phlorotannins, and pigments. Fucoxanthin (Fx) is the most abundant pigment present in brown algae and has shown several useful bioactivities that can be used to fortify products in the food and cosmetic industries. Nevertheless, to date, there is still insufficient literature reporting on the extraction yield of Fx from U. pinnatifida species from green technologies. In this regard, the present study aims to optimize the extraction conditions to obtain the highest Fx yield from U. pinnatifida through emerging techniques, namely microwave-assisted extraction (MAE) and ultrasound-assisted extraction (UAE). These methods will be compared with the conventional methodologies of heat-assisted extraction (HAE) and Soxhlet-assisted extraction (SAE). According to our results, even though the extraction yield could be slightly higher when using MAE than UAE, the Fx concentration obtained from the alga was double when using UAE. Thus, the Fx ratio in the final extract reached values of 124.39 mg Fx/g E. However, the optimal conditions should also be considered since UAE needed 30 min to perform the extraction, whereas MAE was able to obtain 58.83 mg Fx/g E in only 3 min and 2 bar, meaning less energy expenditure and minimum cost function. To our knowledge, this study obtains the highest concentrations of Fx ever reported (58.83 mg Fx/g E for MAE and 124.39 mg Fx/g E for UAE), with low energy consumption and short times (3.00 min for MAE and 35.16 min for UAE). Any of these results could be selected for further experiments and proposed for industrial scaling-up.


Value-Based Health Care in Undergraduate Medical Education.
Value-based health care (VBHC) is an innovative framework for redesigning care delivery to achieve better outcomes for patients and reduce cost; however, providing students with the skills to understand and engage with these topics is a challenge to medical educators. Here, the authors present a novel, VBHC curriculum integrated into a required course for post-core clerkship students-launched in 2018 at Harvard Medical School and taught in conjunction with Harvard Business School faculty-that highlights key principles of VBHC most relevant to undergraduate medical education. The course integrates VBHC with related health disciplines, including health policy, ethics, epidemiology, and social medicine, using a case-based method. Students practice active decision making while learning key concepts to address value in clinical practice. Since the course's inception in March 2018, 95 students (87%) completed the standardized course evaluation; the majority said VBHC content and pedagogical style (i.e., case-based learning) enhanced their learning. Students' critiques focused on too little integration with other disciplines (e.g., social medicine, ethics), the physical space, and inadequate time for debates about potential tensions between VBHC and other course disciplines. The authors believe that by exposing medical students to the principles of VBHC, students will fulfill the expectations of graduating physicians by excelling as critical thinkers, collaborative team members, and judicious care providers throughout their residency, clinical practice, and beyond. Future VBHC curricula expansions may include elective coursework, intensive seminar series, and formal dual degrees.


Interprofessional education for students of the health professions: the "Seamless Care" model.
"Seamless Care" was one of 21 grants awarded by Health Canada to inform policymakers of the effectiveness of interprofessional education in promoting collaborative patient-centred practice among health professionals. The "Seamless Care" model of interprofessional education was designed with input from three Faculties at Dalhousie University (Medicine, Dentistry and Health Professions). The design was grounded in relevant learning theories--Social Cognitive Theory, Self-efficacy, Situated Learning theory and Constructivism. The intervention was informed by principles of active learning, problem-based learning, reflection and role modeling. The primary goal of Seamless Care was to develop students' interprofessional patient-centred collaborative skills through experiential learning. Fourteen student teams, each including one student from medicine, nursing, pharmacy, dentistry and dental hygiene, learned with, from and about each other while they were mentored in the collaborative care of patients transitioning from acute care to the community. Student teams providing collaborative care assisted patients experiencing a chronic illness to become more active in managing their health through development of self-management and decision-making skills. This paper describes the Seamless Care model of interprofessional education and discusses the theoretical underpinnings of this experiential model of interprofessional education designed to extend classroom-based interprofessional education to the clinical setting.


COMT and ALDH2 polymorphisms moderate associations of implicit drinking motives with alcohol use.
Dual process models of addiction emphasize the importance of implicit (automatic) cognitive processes in the development and maintenance of substance use behavior. Although genetic influences are presumed to be relevant for dual process models, few studies have evaluated this possibility. The current study examined two polymorphisms with functional significance for alcohol use behavior (COMT Val158Met and ALDH2*2) in relation to automatic alcohol cognitions and tested additive and interactive effects of genotype and implicit cognitions on drinking behavior. Participants were college students (n = 69) who completed Implicit Association Tasks (IATs) designed to assess two classes of automatic drinking motives (enhancement motives and coping motives). Genetic factors did not show direct associations with IAT measures; however, COMT and ALDH2 moderated associations of implicit coping motives with drinking outcomes. Interaction effects indicated that associations of implicit motives with drinking outcomes were strongest in the context of genetic variants associated with relatively higher risk for alcohol use (COMT Met and ALDH2*1). Associations of genotype with drinking behavior were observed for ALDH2 but not COMT. These findings are consistent with the possibility that genetic risk or protective factors could potentiate or mitigate the influence of reflexive cognitive processes on drinking behavior, providing support for the evaluation of genetic influences in the context of dual process models of addiction.


Nutrigenomic point of view on effects and mechanisms of action of ruminant trans fatty acids on insulin resistance and type 2 diabetes.
Evidence from observational studies suggests beneficial effects of ruminant trans fatty acids (rTFA) on insulin resistance (IR) and type 2 diabetes (T2D). However, beneficial effects of rTFA are not always observed in cell, animal, and human studies. This narrative review presents potential mechanisms of action of rTFA using nutrigenomics and microRNA results in an integrative model. In addition, the review presents factors, including measures of IR and T2D, dose and duration of studies, as well as health status, ethnicity, and genotypes of subjects, that may help explain the heterogeneity in response to rTFA supplementation. Future studies should consider these factors, as well as research in nutritional genomics, to better understand the effects of rTFA on IR and T2D.


Superior immunogenicity of inactivated whole virus H5N1 influenza vaccine is primarily controlled by Toll-like receptor signalling.
In the case of an influenza pandemic, the current global influenza vaccine production capacity will be unable to meet the demand for billions of vaccine doses. The ongoing threat of an H5N1 pandemic therefore urges the development of highly immunogenic, dose-sparing vaccine formulations. In unprimed individuals, inactivated whole virus (WIV) vaccines are more immunogenic and induce protective antibody responses at a lower antigen dose than other formulations like split virus (SV) or subunit (SU) vaccines. The reason for this discrepancy in immunogenicity is a long-standing enigma. Here, we show that stimulation of Toll-like receptors (TLRs) of the innate immune system, in particular stimulation of TLR7, by H5N1 WIV vaccine is the prime determinant of the greater magnitude and Th1 polarization of the WIV-induced immune response, as compared to SV- or SU-induced responses. This TLR dependency largely explains the relative loss of immunogenicity in SV and SU vaccines. The natural pathogen-associated molecular pattern (PAMP) recognized by TLR7 is viral genomic ssRNA. Processing of whole virus particles into SV or SU vaccines destroys the integrity of the viral particle and leaves the viral RNA prone to degradation or involves its active removal. Our results show for a classic vaccine that the acquired immune response evoked by vaccination can be enhanced and steered by the innate immune system, which is triggered by interaction of an intrinsic vaccine component with a pattern recognition receptor (PRR). The insights presented here may be used to further improve the immune-stimulatory and dose-sparing properties of classic influenza vaccine formulations such as WIV, and will facilitate the development of new, even more powerful vaccines to face the next influenza pandemic.


A better self-efficacy is predictive of better health-related quality of life (HRQoL) in patients with systemic lupus erythematosus: data from the Almenara Lupus Cohort.
To determine the possible predictive value of self-efficacy on health-related quality of life (HRQoL) in patients with SLE. Patients with SLE from the Almenara Lupus Cohort were included. Self-efficacy was ascertained with the six domains from the Patient-Reported Outcomes Measurement Information System (PROMIS) self-efficacy for managing chronic conditions. For PROMIS domains, a score of 50 is the average for a clinical population (people with a chronic condition), a higher score indicates that the respondent has greater self-efficacy. HRQoL was ascertained with the physical and mental component summary (PCS and MCS) measures of the Short-Form 36 (SF-36). Generalised estimating equations were performed, using as outcome the PCS or MCS in the subsequent visit, and the self-efficacy domain in the previous visit; multivariable models were adjusted for possible confounders. The confounders were measured in the same visit as the self-efficacy domain. Two-hundred and nine patients for a total of 564 visits were included; 194 (92.8%) patients were women and mean age at diagnosis was 36.4 (14.0) years. In the multivariable models, a better PCS was predicted by a better self-efficacy for managing symptoms, managing medications and treatments and managing social interactions and general self-efficacy; a better MCS was predicted by a better self-efficacy for managing daily activities, managing symptoms, managing medications and treatments and managing social interactions. A better self-efficacy is predictive of subsequent better HRQoL, even after adjustment for possible confounders. These results should encourage clinicians to develop strategies to improve self-efficacy in patients with SLE.


Promoting Healthy Childhood Behaviors With Financial Incentives: A Narrative Review of Key Considerations and Design Features for Future Research.
In the last decade, there has been a robust increase in research using financial incentives to promote healthy behaviors as behavioral economics and new monitoring technologies have been applied to health behaviors. Most studies of financial incentives on health behaviors have focused on adults, yet many unhealthy adult behaviors have roots in childhood and adolescence. The use of financial incentives is an attractive but controversial strategy in childhood. In this review, we first propose 5 general considerations in designing and applying incentive interventions to children. These include: 1) the potential impact of incentives on intrinsic motivation, 2) ethical concerns about incentives promoting undue influence, 3) the importance of child neurodevelopmental stage, 4) how incentive interventions may influence health disparities, and 5) how to finance effective programs. We then highlight empirical findings from randomized trials investigating key design features of financial incentive interventions, including framing (loss vs gain), timing (immediate vs delayed), and magnitude (incentive size) effects on a range of childhood behaviors from healthy eating to adherence to glycemic control in type 1 diabetes. Though the current research base on these subjects in children is limited, we found no evidence suggesting that loss-framed incentives perform better than gain-framed incentives in children and isolated studies from healthy food choice experiments support the use of immediate, small incentives versus delayed, larger incentives. Future research on childhood incentives should compare the effectiveness of gain versus loss-framing and focus on which intervention characteristics lead to sustained behavior change and habit formation.


Screening for sleep-related breathing disorders by transthoracic impedance recording integrated into a Holter ECG system.
In patients with arrhythmias, coincidence with sleep-related breathing disorders (SRBD) is high and of clinical relevance. Electrocardiogram-derived (ECG) parameters have been developed for SRBD screening, but it has proved necessary to exclude patients with frequent arrhythmias. Holter-based screening tools, easy to use, are therefore warranted. The goal of our study was to evaluate the diagnostic accuracy, with respect to SRBD detection, of transthoracic impedance recording (TTIR) integrated into a Holter System. Our investigation consisted of 2 phases. In phase 1 we compared the performance of TTIR to that of in-hospital polysomnography (PSG) in 56 patients (46 male, mean age 57). In phase 2 we compared TTIR to results from an ambulatory polygraphy (PG) system in 180 patients (143 male, mean age 56). We scored apnea and hypopnea from P(S)G, and derived a respiratory-disturbance index (P(S)G-RDI). TTIR was analyzed semi-automatically. Reduction of the impedance amplitude by more than 50% over 10 s was scored as apnea/hypopnea, with consequent calculation of TTIR-RDI. In phase 1, 20 out of 56 patients revealed a PSG-RDI > 10 h(-1). TTIR-RDI in 19 patients from this group was >10 h(-1) (sensitivity 95%, specificity 97.2%, positive predictive value 95%, negative predictive value 97.2%, interclass correlation coefficient 0.98). In phase 2, 46 of 180 patients revealed a PSG-RDI > 10 h(-1). TTIR-RDI in 37 out of this group was >10 h(-1) (sensitivity 80.4%, specificity 92.5%, positive predictive value 78.7%, negative predictive value 93.2%, interclass correlation coefficient 0.92). TTIR integrated into a Holter ECG system and tested in a large patient cohort demonstrates acceptable high accuracy in detection of SRBD. Arrhythmia analysis and screening for SRBD can be performed in a single-step approch.


Efficient acute and chronic infection of stem cell-derived hepatocytes by hepatitis C virus.
Human stem cell-derived hepatocyte-like cells (HLCs) have shown high potential as authentic model for dissection of the HCV life cycle and virus-induced pathogenesis. However, modest HCV replication, possibly due to robust innate immune responses, limits their broader use. To overcome these limitations and to dissect the mechanisms responsible for control of HCV, we analysed expression of key components of the interferon (IFN) system in HLCs, assessed permissiveness for different HCV strains and blocked innate immune signalling by pharmacological intervention. Transcriptional profiling revealed that HLCs constitutively express messenger RNA of RLRs, and members of the IFN pathway. Moreover, HLCs upregulated IFNs and canonical interferon-regulated genes (IRGs) upon transfection with the double-stranded RNA mimic poly(I:C). Infection of HLCs with Jc1-HCVcc produced only limited viral progeny. In contrast, infection with p100, a Jc1-derived virus population with enhanced replication fitness and partial resistance to IFN, resulted in robust yet transient viraemia. Viral titres declined concomitant with a peak of IRG induction. Addition of ruxolitinib, a JAK/STAT inhibitor, permitted chronic infection and raised p100 infectious virus titres to 1×105 FFU/mL. IRGs expression profiling in infected HLCs revealed a landscape of HCV-dependent transcriptional changes similar to HCV-infected primary human hepatocytes, but distinct from Huh-7.5 cells. Withdrawal of ruxolitinib restored innate immune responses and resulted in HCV clearance. This authentic human cell model is well suited to examine acute and chronic host-HCV interactions, particularly IFN-triggered antiviral effector functions and mechanisms of innate immune control of HCV infection.


Antimicrobial activity of red wine and oenological extracts against periodontal pathogens in a validated oral biofilm model.
Previous research findings support an antimicrobial effect of polyphenols against a variety of pathogens, but there is no evidence of this effect against periodontal pathogens in complex biofilms. The purpose of this study was to evaluate the antimicrobial activity of red wine and oenological extracts, rich in polyphenols, against the periodontal pathogens Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum and total bacteria growing in an in vitro oral biofilm static model. A previously validated biofilm model, including Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, F. nucleatum, P. gingivalis and A. actinomycetemcomitans was developed on sterile hydroxyapatite discs. Red wine (and dealcoholized wine), and two polyphenols-rich extracts (from wine and grape seeds) were applied to 72 h biofilms by dipping the discs during 1 and 5 min in the wine solutions and during 30 s and 1 min in the oenological extracts. Resulting biofilms were analyzed by confocal laser scanning microscopy and viable bacteria (colony forming units/mL) were measured by quantitative polymerase chain reaction combined with propidium monoazide. A generalized linear model was constructed to determine the effect of the tested products on the viable bacterial counts of A. actinomycetemcomitans, P. gingivalis and F. nucleatum, as well on the total number of viable bacteria. The results showed that red wine and dealcoholized red wine caused reduction in viability of total bacteria within the biofilm, with statistically significant reductions in the number of viable P. gingivalis after 1 min (p = 0.008) and in A. actinomycetemcomitans after 5 min of exposure (p = 0.011) with red wine. No evidence of relevant antibacterial effect was observed with the oenological extracts, with statistically significant reductions of F. nucleatum after 30 s of exposure to both oenological extracts (p = 0.001). Although moderate, the antimicrobial impact observed in the total bacterial counts and counts of A. actinomycetemcomitans, P. gingivalis and F. nucleatum, encourage further investigations on the potential use of these natural products in the prevention and treatment of periodontal diseases.


Molecular interactions between the specialist herbivore Manduca sexta (lepidoptera, sphingidae) and its natural host Nicotiana attenuata: V. microarray analysis and further characterization of large-scale changes in herbivore-induced mRNAs.
We extend our analysis of the transcriptional reorganization that occurs when the native tobacco, Nicotiana attenuata, is attacked by Manduca sexta larvae by cloning 115 transcripts by mRNA differential display reverse transcription-polymerase chain reaction and subtractive hybridization using magnetic beads (SHMB) from the M. sexta-responsive transcriptome. These transcripts were spotted as cDNA with eight others, previously confirmed to be differentially regulated by northern analysis on glass slide microarrays, and hybridized with Cy3- and Cy5-labeled probes derived from plants after 2, 6, 12, and 24 h of continuous attack. Microarray analysis proved to be a powerful means of verifying differential expression; 73 of the cloned genes (63%) were differentially regulated (in equal proportions from differential display reverse transcription-polymerase chain reaction and SHMB procedures), and of these, 24 (32%) had similarity to known genes or putative proteins (more from SHMB). The analysis provided insights into the signaling and transcriptional basis of direct and indirect defenses used against herbivores, suggesting simultaneous activation of salicylic acid-, ethylene-, cytokinin-, WRKY-, MYB-, and oxylipin-signaling pathways and implicating terpenoid-, pathogen-, and cell wall-related transcripts in defense responses. These defense responses require resources that could be made available by decreases in four photosynthetic-related transcripts, increases in transcripts associated with protein and nucleotide turnover, and increases in transcripts associated with carbohydrate metabolism. This putative up-regulation of defense-associated and down-regulation of growth-associated transcripts occur against a backdrop of altered transcripts for RNA-binding proteins, putative ATP/ADP translocators, chaperonins, histones, and water channel proteins, responses consistent with a major metabolic reconfiguration that underscores the complexity of response to herbivore attack.


Action of snake venom components on the haemostatic system.
Among the components in snake venom are a number which have profound effects (either stimulatory or inhibitory) on haemostatic mechanisms, including coagulation, fibrinolysis, platelet function and vascular integrity. As a consequence, human victims of snakebite may suffer severe and sometimes fatal haemorrhagic and/or thrombotic sequelae. Many of these venom components have been isolated and their precise mechanisms of action established. Apart from direct fibrinolysins, procoagulants predominate, most of these exerting their effect late in the clotting cascade, activating factor X or prothrombin or directly converting fibrinogen to fibrin. Some of the procoagulants are, or have the potential to be, used as therapeutic agents. Some venom components have been put to use as laboratory reagents for diagnostic purposes or for characterising molecular defects of haemostasis, although because they often have unphysiological actions, results must be interpreted with caution. These and other useful constituents e.g. protein C activator and platelet aggregating agents are discussed.


Clinicians' Views and Experiences with Offering and Returning Results from Exome Sequencing to Parents of Infants with Hearing Loss.
Exome sequencing (ES) is an effective method for identifying the genetic cause of hearing loss in infants diagnosed through newborn hearing screening programs. ES has the potential to be integrated into routine clinical care, yet little is known about the experiences of clinicians offering this test to families. To address this gap, clinicians involved in a clinical study using ES to identify the cause of infants' hearing loss were interviewed to explore their experiences with offering and returning results to parents. Interview transcripts were analysed using inductive content analysis. Twelve clinicians participated: seven genetic counsellors, four clinical geneticists, and one paediatrician. Most clinicians were supportive of offering ES to infants with hearing loss, primarily because results may inform the child's clinical management. However, some expressed concerns, questioning the utility of this information, particularly for isolated hearing loss. Clinicians had differing views regarding the optimal time to offer ES to families; while some felt that families can manage everything at once, others recommended delaying testing until parents have come to terms with their child's diagnosis. These findings show the complexity involved in determining how ES should be offered to families following the diagnosis of a child with hearing loss, particularly with regards to when testing is suggested.


Women's health groups to improve perinatal care in rural Nepal.
BACKGROUND: Neonatal mortality rates are high in rural Nepal where more than 90% of deliveries are in the home. Evidence suggests that death rates can be reduced by interventions at community level. We describe an intervention which aimed to harness the power of community planning and decision making to improve maternal and newborn care in rural Nepal. METHODS: The development of 111 women's groups in a population of 86 704 in Makwanpur district, Nepal is described. The groups, facilitated by local women, were the intervention component of a randomized controlled trial to reduce perinatal and neonatal mortality rates. Through participant observation and analysis of reports, we describe the implementation of this intervention: the community entry process, the facilitation of monthly meetings through a participatory action cycle of problem identification, community planning, and implementation and evaluation of strategies to tackle the identified problems. RESULTS: In response to the needs of the group, participatory health education was added to the intervention and the women's groups developed varied strategies to tackle problems of maternal and newborn care: establishing mother and child health funds, producing clean home delivery kits and operating stretcher schemes. Close linkages with community leaders and community health workers improved strategy implementation. There were also indications of positive effects on group members and health services, and most groups remained active after 30 months. CONCLUSION: A large scale and potentially sustainable participatory intervention with women's groups, which focused on pregnancy, childbirth and the newborn period, resulted in innovative strategies identified by local communities to tackle perinatal care problems.


Autophagy inhibition enhances RAD001-induced cytotoxicity in human bladder cancer cells.
Mammalian target of rapamycin (mTOR), involved in PI3K/AKT/mTOR pathway, is known to play a central role in regulating the growth of cancer cells. The PI3K/AKT/mTOR pathway enhances tumor survival and proliferation through suppressing autophagy, which sustains energy homeostasis by collecting and recycling cellular components under stress conditions. Conversely, inhibitors of the mTOR pathway such as RAD001 induce autophagy, leading to promotion of tumor survival and limited antitumor efficacy. We thus hypothesized that the use of autophagy inhibitor in combination with mTOR inhibition improves the cytotoxicity of mTOR inhibitors in bladder cancer. The cytotoxicity of RT4, 5637, HT1376, and T24 human bladder cancer cells treated with RAD001 alone or combined with autophagy inhibitors (3-methyladenine (3-MA), bafilomycin A1 (Baf A1), chloroquine, or hydroxychloroquine) was assessed using the WST-8 cell viability kit. The autophagy status in cells was analyzed by the detection of microtubule-associated light chain 3 form II (LC3-II), using immunofluorescent staining and Western blot. Acidic vesicular organelle (AVO) formation in treated cells was determined by acridine orange vital staining. Inhibition of mTOR pathway by RAD001 was monitored by using a homemade quantitative polymerase chain reaction gene array, while phospho-mTOR was detected using Western blot. Induced apoptosis was determined by measurement of caspase 3/7 activity and DNA fragmentation in cells after treatment. Advanced bladder cancer cells (5637, HT1376, and T24) were more resistant to RAD001 than RT4. Autophagy flux detected by the expression of LC3-II showed RAD001-induced autophagy. AVO formation was detected in cells treated with RAD001 and was inhibited by the addition of 3-MA or Baf A1. Cotreatment of RAD001 with autophagy inhibitors further reduced cell viability and induced apoptosis in bladder cancer cells. Our results indicate that simultaneous inhibition of the mTOR and autophagy pathway significantly enhances apoptosis, and it is suggested to be a new therapeutic paradigm for the treatment of bladder cancer.