Identification of natural and artificial DNA substrates for light-activated LOV-HTH transcription factor EL222.
Light-oxygen-voltage (LOV) domains serve as the photosensory modules for a wide range of plant and bacterial proteins, conferring blue light-dependent regulation to effector activities as diverse as enzymes and DNA binding. LOV domains can also be engineered into a variety of exogenous targets, allowing similar regulation for new protein-based reagents. Common to these proteins is the ability for LOV domains to reversibly form a photochemical adduct between an internal flavin chromophore and the surrounding protein, using this to trigger conformational changes that affect output activity. Using the Erythrobacter litoralis protein EL222 model system that links LOV regulation to a helix-turn-helix (HTH) DNA binding domain, we demonstrated that the LOV domain binds and inhibits the HTH domain in the dark, releasing these interactions upon illumination [Nash, A. I., et al. (2011) Proc. Natl. Acad. Sci. U.S.A. 108, 9449-9454]. Here we combine genomic and in vitro selection approaches to identify optimal DNA binding sites for EL222. Within the bacterial host, we observe binding at several genomic sites using a 12 bp sequence consensus that is also found by in vitro selection methods. Sequence-specific alterations in the DNA consensus reduce EL222 binding affinity in a manner consistent with the expected binding mode, a protein dimer binding to two repeats. Finally, we demonstrate the light-dependent activation of transcription of two genes adjacent to an EL222 binding site. Taken together, these results shed light on the native function of EL222 and provide useful reagents for further basic and applications research of this versatile protein.


Applying the cognitive theory of multimedia learning: an analysis of medical animations.
Instructional animations play a prominent role in medical education, but the degree to which these teaching tools follow empirically established learning principles, such as those outlined in the cognitive theory of multimedia learning (CTML), is unknown. These principles provide guidelines for designing animations in a way that promotes optimal cognitive processing and facilitates learning, but the application of these learning principles in current animations has not yet been investigated. A large-scale review of existing educational tools in the context of this theoretical framework is necessary to examine if and how instructional medical animations adhere to these principles and where improvements can be made. We conducted a comprehensive review of instructional animations in the health sciences domain and examined whether these animations met the three main goals of CTML: managing essential processing; minimising extraneous processing, and facilitating generative processing. We also identified areas for pedagogical improvement. Through Google keyword searches, we identified 4455 medical animations for review. After the application of exclusion criteria, 860 animations from 20 developers were retained. We randomly sampled and reviewed 50% of the identified animations. Many animations did not follow the recommended multimedia learning principles, particularly those that support the management of essential processing. We also noted an excess of extraneous visual and auditory elements and few opportunities for learner interactivity. Many unrealised opportunities exist for improving the efficacy of animations as learning tools in medical education; instructors can look to effective examples to select or design animations that incorporate the established principles of CTML.


CYP7A1, BAAT and UGT1A1 polymorphisms and susceptibility to anti-tuberculosis drug-induced hepatotoxicity.
Evidence indicates that the polymorphisms in genes involved in bile acid metabolism may play an important role in the development of anti-tuberculosis drug-induced hepatotoxicity (ATDH) in tuberculosis (TB) patients treated with anti-tuberculosis drugs. To investigate the association between genetic variants of CYP7A1, BAAT and UGT1A1 and the risk of ATDH in a Chinese cohort. In this nested case-control study, 89 TB patients with ATDH and 356 matched ATDH-free TB patients constituted cases and controls, respectively. Genetic polymorphisms of CYP7A1, BAAT and UGT1A1 were determined using the TaqMan single-nucleotide polymorphism genotyping assay. Odds ratios (ORs) with 95% confidence intervals (CIs) were estimated using a conditional logistic regression model. Significant differences were found in genotype distributions of rs1457043 in CYP7A1 between patients with and those without ATDH (P = 0.014). Genotype and haplotype analysis showed that patients carrying an AG genotype of rs1457043 and G-C or G-A haplotypes of rs1457043-rs8192870 in CYP7A1 were at a higher risk of ATDH than those with GG genotype and A-C haplotype, with ORs of respectively 2.05 (95%CI 1.18-3.15) and 2.40 (95%CI 1.62-3.57). Genetic polymorphisms of CYP7A1 may be associated with susceptibility to ATDH in the Chinese population.


Effects of approach and injection volume on diffusion of mepivacaine hydrochloride during local analgesia of the deep branch of the lateral plantar nerve in horses.
To compare the effects of 2 approaches and 2 injection volumes on diffusion of mepivacaine hydrochloride for local analgesia of the deep branch of the lateral plantar nerve (DBLPN) in horses. Experimental study. 16 adult horses. Either 2 mL (low volume) or 8 mL (high volume) of mepivacaine hydrochloride-iohexol (50:50 mixture) was injected by means of 1 of 2 techniques to produce analgesia of the DBLPN. For technique 1, the needle was inserted 15 mm distal to the head of the fourth metatarsal bone and directed perpendicular to the limb. For technique 2, the needle was inserted 20 mm distal to the head of the fourth metatarsal bone and was directed in a proximodorsal direction. Lateromedial radiographs were obtained before and 5, 15, 30, and 60 minutes after injection. Radiographs were evaluated to determine the proximal and distal extent of diffusion of the contrast solution and presumably anesthetic agent and whether contrast agent appeared to be present in the tarsal sheath or tarsometatarsal joint. A high degree of variability in contrast solution diffusion was noted among injections. High-volume injections diffused significantly further proximally and distally than did low-volume injections. Contrast agent was documented within the tarsal sheath in 5 of 32 (16%) injections and within the tarsometatarsal joint in 2 of 32 (6%) injections. No significant difference was found for risk of inadvertent tarsal sheath or tarsometatarsal joint injection between the 2 techniques or the 2 volumes of anesthetic used. Mepivacaine diffused significantly further distally with technique 1 than with technique 2 but diffused significantly further proximally with technique 2 than with technique 1. For both techniques, diffusion in the distal but not the proximal direction significantly increased over time. Results indicated that the proximal and distal diffusion of the mepivacaine-iohexol solution was quite variable following either DBLPN nerve block technique.


Oral Chemotherapy Adherence: A Novel Nursing Intervention Using an Electronic Health Record Workflow .
In the ambulatory care setting, chemotherapy regimens have become increasingly complex with the combination of induction treatments and oral medications. Nurses at one cancer center implemented an oral adherence tracking documentation system in the electronic health record (EHR). Oncology nurses assessed and monitored adherence to oral chemotherapy at each clinical encounter and during telephone calls and then documented findings in the EHR. After implementing this new standardized approach, adherence rates were captured as a metric for the organization.


Tests of the double-strand-break repair model for red-mediated recombination of phage lambda and plasmid lambda dv.
The double-strand-break repair (DSBR) model was formulated to account for various aspects of yeast mitotic and meiotic recombination. In this study three features of the DSBR model are tested for Red-mediated recombination between phage lambda and lambda dv, a plasmid that is perfectly homologous to about 10% of lambda. The results support the applicability of the DSBR model to lambda's Red system: (1) Creating a double-strand-break (DSB) within the region of homology shared by phage and plasmid increases their genetic interaction by about 20-fold. A DSB outside the region of shared homology has no such effect. (2) Both patches, i.e., simple marker rescue, and splices, i.e., co-integration of the phage and plasmid, are stimulated by a DSB in the region of shared homology. (3) Co-integrants harbor a duplication of the region of shared homology. Among co-integrants that were formed by the creation of a DSB, there is a preferential loss of whichever allele was in cis to a utilized cut site. The DSBR model as originally formulated involves the isomerization and cleavage of Holliday junctions to resolve the canonical intermediate. We propose as an alternative mechanism that a topoisomerase can resolve the canonical DSBR intermediate.


Gender-related power differences, beliefs and reactions towards people living with HIV/AIDS: an urban study in Nigeria.
Although there are an increasing number of studies on HIV-related stigma in Nigeria, very little research has focused on how power differences based on gender perpetuate the stigmatization of people living with HIV/AIDS (PLWHA) and how these gender differences affect the care that PLWHA receive in health care institutions. We explore gender-related beliefs and reactions of society, including health care professionals (HCPs), with regard to PLWHA, using Connell's theoretical framework of gender and power (1987). With Connell's structural theory of gender and power (financial inequality, authority and structure of social norms), we can describe gender differences in stigmatization of PLWHA. We conducted in-depth semi-structured interviews, lasting 60 to 90 minutes, with 100 persons (40 members of the general public, 40 HCPs and 20 PLWHA) in Port Harcourt, Nigeria. The interviews were tape-recorded and transcribed verbatim. The Nvivo 7 computer package was used to analyze the data. There are similarities and differences between the general public and HCPs towards PLWHA in gender-related beliefs and reactions. For instance, although association with promiscuity and power differences were commonly acknowledged in the different groups, there are differences in how these reactions are shown; such as HCPs asking the female PLWHA to inform their partners to ensure payment of hospital bills. Women with HIV/AIDS in particular are therefore in a disadvantaged position with regard to the care they receive. Despite the fact that men and women with HIV/AIDS suffer the same illness, clear disparities are apparent in the negative reaction women and men living with HIV/AIDS experience in society. We show that women's generally low status in society contributes to the extreme negative reactions to which female PLWHA are subject. The government should create policies aimed at reducing the power differences in family, society and health care systems, which would be important to decrease the gender-related differences in stigma experienced by PLWHA. Interventions should be directed at the prevailing societal norms through appropriate legislation and advocacy at grassroots level with the support of men to counter laws that put women in a disadvantaged position. Furthermore, development of a policy that encourages equality in access to health care for all patients with HIV/AIDS by applying the same conditions to both men and women in health care institutions is recommended. There is a need to protect women's rights through implementing support policies, including paying attention to gender in the training of HCPs.


Translucent soft robots driven by frameless fluid electrode dielectric elastomer actuators.
Dielectric elastomer actuators (DEAs) are a promising enabling technology for a wide range of emerging applications, including robotics, artificial muscles, and microfluidics. This is due to their large actuation strains, rapid response rate, low cost and low noise, high energy density, and high efficiency when compared with alternative actuators. These properties make DEAs ideal for the actuation of soft submersible devices, although their use has been limited because of three main challenges: (i) developing suitable, compliant electrode materials; (ii) the need to effectively insulate the actuator electrodes from the surrounding fluid; and (iii) the rigid frames typically required to prestrain the dielectric layers. We explored the use of a frameless, submersible DEA design that uses an internal chamber filled with liquid as one of the electrodes and the surrounding environmental liquid as the second electrode, thus simplifying the implementation of soft, actuated submersible devices. We demonstrated the feasibility of this approach with a prototype swimming robot composed of transparent bimorph actuator segments and inspired by transparent eel larvae, leptocephali. This design achieved undulatory swimming with a maximum forward swimming speed of 1.9 millimeters per second and a Froude efficiency of 52%. We also demonstrated the capability for camouflage and display through the body of the robot, which has an average transmittance of 94% across the visible spectrum, similar to a leptocephalus. These results suggest a potential for DEAs with fluid electrodes to serve as artificial muscles for quiet, translucent, swimming soft robots for applications including surveillance and the unobtrusive study of marine life.


Responses of benthic macroinvertebrates to environmental changes associated with urbanization in nine metropolitan areas.
Responses of benthic macroinvertebrates along gradients of urban intensity were investigated in nine metropolitan areas across the United States. Invertebrate assemblages in metropolitan areas where forests or shrublands were being converted to urban land were strongly related to urban intensity. In metropolitan areas where agriculture and grazing lands were being converted to urban land, invertebrate assemblages showed much weaker or nonsignificant relations with urban intensity because sites with low urban intensity were already degraded by agriculture. Ordination scores, the number of EPT taxa, and the mean pollution-tolerance value of organisms at a site were the best indicators of changes in assemblage condition. Diversity indices, functional groups, behavior, and dominance metrics were not good indicators of urbanization. Richness metrics were better indicators of urban effects than were abundance metrics, and qualitative samples collected from multiple habitats gave similar results to those of single habitat quantitative samples (riffles or woody snags) in all metropolitan areas. Changes in urban intensity were strongly correlated with a set of landscape variables that was consistent across all metropolitan areas. In contrast, the instream environmental variables that were strongly correlated with urbanization and invertebrate responses varied among metropolitan areas. The natural environmental setting determined the biological, chemical, and physical instream conditions upon which urbanization acts and dictated the differences in responses to urbanization among metropolitan areas. Threshold analysis showed little evidence for an initial period of resistance to urbanization. Instead, assemblages were degraded at very low levels of urbanization, and response rates were either similar across the gradient or higher at low levels of urbanization. Levels of impervious cover that have been suggested as protective of streams (5-10%) were associated with significant assemblage degradation and were not protective.


Treatment and seroconversion in a cohort of children suffering from recent chronic Chagas infection in Yoro, Honduras.
Between 1999-2002, Médécins Sans Frontières-Spain implemented a project seeking to determine the efficacy and safety of benznidazole in the treatment of recent chronic Chagas disease in a cohort of seropositive children in the Yoro Department, Honduras. A total of 24,471 children were screened for Trypanosoma cruzi IgG antibodies through conventional enzyme-linked immunosorbent assays (ELISA) on filter paper. Recombinant ELISA (0.93% seroprevalence) showed 256 initially reactive cases, including 232 confirmed positive cases. Of these, 231 individuals were treated with benznidazole (7.5 mg/kg/day) for 60 days and were followed with a strict weekly medical control and follow-up protocol. At the end of the project, 229 patients were examined by the Honduras Secretariat of Health for post-treatment serological assessments; 88.2% seroconverted after 18 months and 93.9% seroconverted after three years. No differences were found in the seroconversion rates according to age or sex. Most of the side effects of the treatment were minor. These results support the argument that in areas where T. cruzi I is predominant and in areas affected by T. cruzi II, when vector transmission has been interrupted, Chagas disease diagnosis and treatment are feasible, necessary and ethically indisputable.


A potato model intercomparison across varying climates and productivity levels.
A potato crop multimodel assessment was conducted to quantify variation among models and evaluate responses to climate change. Nine modeling groups simulated agronomic and climatic responses at low-input (Chinoli, Bolivia and Gisozi, Burundi)- and high-input (Jyndevad, Denmark and Washington, United States) management sites. Two calibration stages were explored, partial (P1), where experimental dry matter data were not provided, and full (P2). The median model ensemble response outperformed any single model in terms of replicating observed yield across all locations. Uncertainty in simulated yield decreased from 38% to 20% between P1 and P2. Model uncertainty increased with interannual variability, and predictions for all agronomic variables were significantly different from one model to another (P < 0.001). Uncertainty averaged 15% higher for low- vs. high-input sites, with larger differences observed for evapotranspiration (ET), nitrogen uptake, and water use efficiency as compared to dry matter. A minimum of five partial, or three full, calibrated models was required for an ensemble approach to keep variability below that of common field variation. Model variation was not influenced by change in carbon dioxide (C), but increased as much as 41% and 23% for yield and ET, respectively, as temperature (T) or rainfall (W) moved away from historical levels. Increases in T accounted for the highest amount of uncertainty, suggesting that methods and parameters for T sensitivity represent a considerable unknown among models. Using median model ensemble values, yield increased on average 6% per 100-ppm C, declined 4.6% per °C, and declined 2% for every 10% decrease in rainfall (for nonirrigated sites). Differences in predictions due to model representation of light utilization were significant (P < 0.01). These are the first reported results quantifying uncertainty for tuber/root crops and suggest modeling assessments of climate change impact on potato may be improved using an ensemble approach.


The effect of vitamin E and beta carotene on the incidence of lung cancer and other cancers in male smokers.
Epidemiologic evidence indicates that diets high in carotenoid-rich fruits and vegetables, as well as high serum levels of vitamin E (alpha-tocopherol) and beta carotene, are associated with a reduced risk of lung cancer. We performed a randomized, double-blind, placebo-controlled primary-prevention trial to determine whether daily supplementation with alpha-tocopherol, beta carotene, or both would reduce the incidence of lung cancer and other cancers. A total of 29,133 male smokers 50 to 69 years of age from southwestern Finland were randomly assigned to one of four regimens: alpha-tocopherol (50 mg per day) alone, beta carotene (20 mg per day) alone, both alpha-tocopherol and beta carotene, or placebo. Follow-up continued for five to eight years. Among the 876 new cases of lung cancer diagnosed during the trial, no reduction in incidence was observed among the men who received alpha-tocopherol (change in incidence as compared with those who did not, -2 percent; 95 percent confidence interval, -14 to 12 percent). Unexpectedly, we observed a higher incidence of lung cancer among the men who received beta carotene than among those who did not (change in incidence, 18 percent; 95 percent confidence interval, 3 to 36 percent). We found no evidence of an interaction between alpha-tocopherol and beta carotene with respect to the incidence of lung cancer. Fewer cases of prostate cancer were diagnosed among those who received alpha-tocopherol than among those who did not. Beta carotene had little or no effect on the incidence of cancer other than lung cancer. Alpha-tocopherol had no apparent effect on total mortality, although more deaths from hemorrhagic stroke were observed among the men who received this supplement than among those who did not. Total mortality was 8 percent higher (95 percent confidence interval, 1 to 16 percent) among the participants who received beta carotene than among those who did not, primarily because there were more deaths from lung cancer and ischemic heart disease. We found no reduction in the incidence of lung cancer among male smokers after five to eight years of dietary supplementation with alpha-tocopherol or beta carotene. In fact, this trial raises the possibility that these supplements may actually have harmful as well as beneficial effects.


Off-target effects of psychoactive drugs revealed by genome-wide assays in yeast.
To better understand off-target effects of widely prescribed psychoactive drugs, we performed a comprehensive series of chemogenomic screens using the budding yeast Saccharomyces cerevisiae as a model system. Because the known human targets of these drugs do not exist in yeast, we could employ the yeast gene deletion collections and parallel fitness profiling to explore potential off-target effects in a genome-wide manner. Among 214 tested, documented psychoactive drugs, we identified 81 compounds that inhibited wild-type yeast growth and were thus selected for genome-wide fitness profiling. Many of these drugs had a propensity to affect multiple cellular functions. The sensitivity profiles of half of the analyzed drugs were enriched for core cellular processes such as secretion, protein folding, RNA processing, and chromatin structure. Interestingly, fluoxetine (Prozac) interfered with establishment of cell polarity, cyproheptadine (Periactin) targeted essential genes with chromatin-remodeling roles, while paroxetine (Paxil) interfered with essential RNA metabolism genes, suggesting potential secondary drug targets. We also found that the more recently developed atypical antipsychotic clozapine (Clozaril) had no fewer off-target effects in yeast than the typical antipsychotics haloperidol (Haldol) and pimozide (Orap). Our results suggest that model organism pharmacogenetic studies provide a rational foundation for understanding the off-target effects of clinically important psychoactive agents and suggest a rational means both for devising compound derivatives with fewer side effects and for tailoring drug treatment to individual patient genotypes.


Molecular and Epigenetic Mechanisms of MLL in Human Leukemogenesis.
Epigenetics is often defined as the study of heritable changes in gene expression or chromosome stability that don't alter the underlying DNA sequence. Epigenetic changes are established through multiple mechanisms that include DNA methylation, non-coding RNAs and the covalent modification of specific residues on histone proteins. It is becoming clear not only that aberrant epigenetic changes are common in many human diseases such as leukemia, but that these changes by their very nature are malleable, and thus are amenable to treatment. Epigenetic based therapies have so far focused on the use of histone deacetylase (HDAC) inhibitors and DNA methyltransferase inhibitors, which tend to have more general and widespread effects on gene regulation in the cell. However, if a unique molecular pathway can be identified, diseases caused by epigenetic mechanisms are excellent candidates for the development of more targeted therapies that focus on specific gene targets, individual binding domains, or specific enzymatic activities. Designing effective targeted therapies depends on a clear understanding of the role of epigenetic mutations during disease progression. The Mixed Lineage Leukemia (MLL) protein is an example of a developmentally important protein that controls the epigenetic activation of gene targets in part by methylating histone 3 on lysine 4. MLL is required for normal development, but is also mutated in a subset of aggressive human leukemias and thus provides a useful model for studying the link between epigenetic cell memory and human disease. The most common MLL mutations are chromosome translocations that fuse the MLL gene in frame with partner genes creating novel fusion proteins. In this review, we summarize recent work that argues MLL fusion proteins could function through a single molecular pathway, but we also highlight important data that suggests instead that multiple independent mechanisms underlie MLL mediated leukemogenesis.


Determinants of underdiagnosis of COPD in national and international surveys.
COPD ranks within the top three causes of mortality in the global burden of disease, yet it remains largely underdiagnosed. We assessed the underdiagnosis of COPD and its determinants in national and international surveys of general populations. We analyzed representative samples of adults aged ≥ 40 years randomly selected from well-defined administrative areas worldwide (44 sites from 27 countries). Postbronchodilator FEV1/FVC < lower limit of normal (LLN) was used to define chronic airflow limitation consistent with COPD. Undiagnosed COPD was considered when participants had postbronchodilator FEV1/FVC < LLN but were not given a diagnosis of COPD. Among 30,874 participants with a mean age of 56 years, 55.8% were women, and 22.9% were current smokers. Population prevalence of (spirometrically defined) COPD ranged from 3.6% in Barranquilla, Colombia, to 19.0% in Cape Town, South Africa. Only 26.4% reported a previous lung function test, and only 5.0% reported a previous diagnosis of COPD, whereas 9.7% had a postbronchodilator FEV1/FVC < LLN. Overall, 81.4% of (spirometrically defined) COPD cases were undiagnosed, with the highest rate in Ile-Ife, Nigeria (98.3%) and the lowest rate in Lexington, Kentucky (50.0%). In multivariate analysis, a greater probability of underdiagnosis of COPD was associated with male sex, younger age, never and current smoking, lower education, no previous spirometry, and less severe airflow limitation. Even with substantial heterogeneity in COPD prevalence, COPD underdiagnosis is universally high. Because effective management strategies are available for COPD, spirometry can help in the diagnosis of COPD at a stage when treatment will lead to better outcomes and improved quality of life.


Microbial players and processes involved in phytoplankton bloom utilization in the water column of a fast-flowing, river-dominated estuary.
Fueled by seasonal phytoplankton blooms, the Columbia River estuary is a natural bioreactor for organic matter transformations. Prior metagenome analyses indicated high abundances of diverse Bacteroidetes taxa in estuarine samples containing phytoplankton. To examine the hypothesis that Bacteroidetes taxa have important roles in phytoplankton turnover, we further analyzed metagenomes from water collected along a salinity gradient at 0, 5, 15, 25, and 33 PSU during bloom events. Size fractions were obtained by using a 3-μm prefilter and 0.2-μm collection filter. Although this approach targeted bacteria by removing comparatively large eukaryotic cells, the metagenome from the ES-5 sample (5 PSU) nevertheless contained an abundance of diatom DNA. Biogeochemical measurements and prior studies indicated that this finding resulted from the leakage of cellular material due to freshwater diatom lysis at low salinity. Relative to the other metagenomes, the bacterial fraction of ES-5 was dramatically depleted of genes annotated as Bacteroidetes and lysogenic bacteriophages, but was overrepresented in DNA of protists and Myxococcales bacterivores. We suggest the following equally plausible scenarios for the microbial response to phytoplankton lysis: (1) Bacteroidetes depletion in the free-living fraction may at least in part be caused by their attachment to fluvial diatoms as the latter are lysed upon contact with low-salinity estuarine waters; (2) diatom particle colonization is likely followed by rapid bacterial growth and lytic phage infection, resulting in depletion of lysogenic bacteriophages and host bacteria; and (3) the subsequent availability of labile organic matter attracted both grazers and predators to feed in this estuarine biogeochemical "hotspot," which may have additionally depleted Bacteroidetes populations. These results represent the first detailed molecular analysis of the microbial response to phytoplankton lysis at the freshwater-brackish water interface in the fast-flowing Columbia River estuary.


Hepatitis C virus (HCV) genotypes and chronic liver disease in Pakistan.
Hepatitis C virus (HCV) is classified into different types depending on nucleotide sequence variability. Detailed information on the distribution of various HCV genotypes in some geographical areas is available but little is known about Pakistan. In this study, a 5' non-coding region (NCR)-based restriction fragment length polymorphism (RFLP) genotyping assay was used to investigate the genotype distribution in a large series of HCV-infected patients in Karachi, Pakistan. Serum samples from 74 hepatitis B surface antigen (HBsAg)-negative patients with a clinical diagnosis of chronic liver disease (60 patients) and hepatocellular carcinoma (HCC) (14 patients) were assayed for anti-HCV antibody by second generation enzyme immunoassay and 48 were confirmed anti-HCV-positive (33 males, 15 females). Other causes of chronic liver disease (e.g. haemochromatosis, Wilson's disease and immune-mediated injury) were ruled out. Liver biopsy was done in 27/48 anti-HCV-positive patients and in all HCC patients. Genotypes were determined for 45/48 anti-HCV-positive study patients; 39/45 (87%) were type 3; four (9%) were type 1; one was type 2; and one was type 5. Past blood transfusion was the main identifiable risk factor found in 10 patients, all type 3. Seven of the 14 HCC patients were anti-HCV positive, (six were type 3). Most patients with hepatitis C presented with established cirrhosis and complications of portal hypertension and liver failure. (i) genotype 3 is the most common isolate in HCV-associated chronic liver disease in Pakistan; (ii) a significant proportion of HBsAg-negative cirrhotics are non-B, non-C in aetiology; and (iii) half of the patients with HCC have serological evidence of HCV infection.


[Premenstrual dysphoric disorder is often overlooked].
Premenstrual dysphoric disorder (PMDD) is a cluster of severe affective symptoms recurring during the luteal phase of the menstrual cycle, and it affects up to 8% of menstruating women. In Denmark, this disorder is often overlooked. Unlike in the treatment of anxiety and depression, selective serotonin reuptake inhibitors need not be given continuously but can be effective when used only in the luteal phase. This review provides an overview of the current knowledge on the diagnosis and treatment of PMDD in Denmark.


Socioeconomic status and alcohol use in low- and lower-middle income countries: A systematic review.
Harmful use of alcohol is a major cause of global morbidity and mortality. The role of alcohol as a driver of the unfolding non-communicable disease crisis has led to high-profile calls for better epidemiological data. Despite causing a disproportionate amount of harm in low-income groups, there is a critical dearth of evidence on the intra-national socioeconomic patterning of alcohol use in low- and lower-middle income countries (LLMICs). This review aims to fill the gap, providing evidence on the association between socioeconomic status (SES) and alcohol use in these low-income settings. We conducted a comprehensive literature search for primary research published between January 1, 1990 and June 30, 2015 using 13 electronic databases, including Embase and Medline. We also hand-searched references and reviewed 'gray literature' - studies that have not been published in peer-reviewed journals. We included studies from LLMICs presenting data on multiple measures of socioeconomic status and alcohol use. No age or language restrictions were applied. Due to high heterogeneity, we used a narrative approach for data synthesis. After reviewing 4242 records and 247 full-text articles, 23 studies met our inclusion criteria, reporting data on 861,295 individuals aged >10 years from 10 countries. Alcohol use was found to be more prevalent in lower socioeconomic groups in the majority of Southeast Asian studies. The association was mixed for African studies, although these tended to have smaller sample sizes and weaker methods. Studies that measured multiple domains of SES found good agreement between different indicators. Definitions of alcohol use and abuse varied widely between studies, as did socioeconomic groupings. The lack of consistency between studies and the abject lack of data from the majority of LLMICs present a major barrier to policymakers tasked with reducing alcohol-related harm in these settings. Adherence to standardized definitions, the publication of WHO survey data on alcohol and SES, and enhanced surveillance is needed to build an accurate picture of the socioeconomic patterning of alcohol use in developing countries.


Prevalence of the K65R resistance reverse transcriptase mutation in different HIV-1 subtypes in Israel.
The K65R mutation in HIV-1 reverse transcriptase (RT) can be selected by the RT inhibitors tenofovir (TDF), abacavir (ABC), and didanosine (DDI). Recently, in vitro studies have shown that K65R is selected in tissue culture more rapidly with subtype C than subtype B viruses. The prevalence of K65R in viruses sequenced at the Tel-Aviv AIDS Center was evaluated. This study analyzed retrospectively sequences from 1999 to 2007 in patients treated with TDF, ABC, and/or DDI and compared rates of mutational prevalence between subtypes. Fisher's exact test was used to determine statistical significance. Forty-four sequences from patients treated with the three above-cited drugs were analyzed. Subtypes A (n = 1), CRF01_AE (n = 4), CRF02_AG (n = 2), B (n = 21), C (n = 11), D (n = 1), F (n = 3), and G (n = 1) were represented. Seven non-B viruses had the K65R mutation, which was only found in one subtype B virus. Of these seven samples four were subtype C, one was subtype CRF01_AE, and two were subtype CRF02_AG. None of the eight viruses with K65R harbored thymidine analogue mutations. In this study, non-subtype B viruses possessed the K65R mutation at higher incidence than subtype B viruses. Subtype C viruses may be especially prone to develop this mutation. Larger studies are needed to confirm these data. Efforts should be intensified to understand better differences in drug resistance between various HIV subtypes.


Adjuvant immunotherapy with autologous cytokine-induced killer cells for hepatocellular carcinoma.
No adjuvant therapy has been shown to extend the survival of patients with hepatocellular carcinoma (HCC) receiving curative treatment. We investigated whether injections of activated cytokine-induced killer (CIK) cells (CD3+/CD56+ and CD3+/CD56- T cells and CD3-/CD56+ natural killer cells) prolongs recurrence-free survival of patients after curative therapy for HCC. We performed a multicenter, randomized, open-label, phase 3 trial of the efficacy and safety of adjuvant immunotherapy with activated CIK cells (created by incubation of patients' peripheral blood mononuclear cells with interleukin 2 and an antibody against CD3). The study included 230 patients with HCC treated by surgical resection, radiofrequency ablation, or percutaneous ethanol injection at university-affiliated hospitals in Korea. Patients were assigned randomly to receive immunotherapy (injection of 6.4 × 10(9) autologous CIK cells, 16 times during 60 weeks) or no adjuvant therapy (controls). The primary end point was recurrence-free survival; secondary end points included overall survival, cancer-specific survival, and safety. The median time of recurrence-free survival was 44.0 months in the immunotherapy group and 30.0 months in the control group (hazard ratio with immunotherapy, 0.63; 95% confidence interval [CI], 0.43-0.94; P = .010 by 1-sided log-rank test). Hazard ratios also were lower in the immunotherapy than in the control group for all-cause death (0.21; 95% CI, 0.06-0.75; P = .008) and cancer-related death (0.19; 95% CI, 0.04-0.87; P = .02). A significantly higher proportion of patients in the immunotherapy group than in the control group had an adverse event (62% vs 41%; P = .002), but the proportion of patients with serious adverse events did not differ significantly between groups (7.8% vs 3.5%; P = .15). In patients who underwent curative treatment for HCC, adjuvant immunotherapy with activated CIK cells increased recurrence-free and overall survival. ClinicalTrials.gov number: NCT00699816.


MRgFUS Pallidothalamic Tractotomy for Chronic Therapy-Resistant Parkinson's Disease in 51 Consecutive Patients: Single Center Experience.
Background: There is a long history, beginning in the 1940s, of ablative neurosurgery on the pallidal efferent fibers to treat patients suffering from Parkinson's disease (PD). Since the early 1990s, we undertook a re-actualization of the approach to the subthalamic region, and proposed, on a histological basis, to target specifically the pallidothalamic tract at the level of Forel's field H1. This intervention, the pallidothalamic tractotomy (PTT), has been performed since 2011 using the MR-guided focused ultrasound (MRgFUS) technique. A reappraisal of the histology of the pallidothalamic tract was combined recently with an optimization of our lesioning strategy using thermal dose control. Objective: This study was aimed at demonstrating the efficacy and risk profile of MRgFUS PTT against chronic therapy-resistant PD. Methods: This consecutive case series reflects our current treatment routine and was collected between 2017 and 2018. Fifty-two interventions in 47 patients were included. Fifteen patients received bilateral PTT. The median follow-up was 12 months. Results: The Unified Parkinson's Disease Rating Scale (UPDRS) off-medication postoperative score was compared to the baseline on-medication score and revealed percentage reductions of the mean of 84% for tremor, 70% for rigidity, and 73% for distal hypobradykinesia, all values given for the treated side. Axial items (for voice, trunk and gait) were not significantly improved. PTT achieved 100% suppression of on-medication dyskinesias as well as reduction in pain (p < 0.001), dystonia (p < 0.001) and REM sleep disorders (p < 0.01). Reduction of the mean L-Dopa intake was 55%. Patients reported an 88% mean tremor relief and 82% mean global symptom relief on the operated side and 69% mean global symptom improvement for the whole body. There was no significant change of cognitive functions. The small group of bilateral PTTs at 1 year follow-up shows similar results as compared to unilateral PTTs but does not allow to draw firm conclusions at this point. Conclusion: MRgFUS PTT was shown to be a safe and effective intervention for PD patients, addressing all symptoms, with varying effectiveness. We discuss the need to integrate the preoperative state of the thalamocortical network as well as the psycho-emotional dimension.


Mineralogy at Meridiani Planum from the Mini-TES Experiment on the Opportunity Rover.
The Miniature Thermal Emission Spectrometer (Mini-TES) on Opportunity investigated the mineral abundances and compositions of outcrops, rocks, and soils at Meridiani Planum. Coarse crystalline hematite and olivine-rich basaltic sands were observed as predicted from orbital TES spectroscopy. Outcrops of aqueous origin are composed of 15 to 35% by volume magnesium and calcium sulfates [a high-silica component modeled as a combination of glass, feldspar, and sheet silicates (approximately 20 to 30%)], and hematite; only minor jarosite is identified in Mini-TES spectra. Mini-TES spectra show only a hematite signature in the millimeter-sized spherules. Basaltic materials have more plagioclase than pyroxene, contain olivine, and are similar in inferred mineral composition to basalt mapped from orbit. Bounce rock is dominated by clinopyroxene and is close in inferred mineral composition to the basaltic martian meteorites. Bright wind streak material matches global dust. Waterlain rocks covered by unaltered basaltic sands suggest a change from an aqueous environment to one dominated by physical weathering.


A computational model of induced pluripotent stem-cell derived cardiomyocytes incorporating experimental variability from multiple data sources.
Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) capture patient-specific genotype-phenotype relationships, as well as cell-to-cell variability of cardiac electrical activity Computational modelling and simulation provide a high throughput approach to reconcile multiple datasets describing physiological variability, and also identify vulnerable parameter regimes We have developed a whole-cell model of iPSC-CMs, composed of single exponential voltage-dependent gating variable rate constants, parameterized to fit experimental iPSC-CM outputs We have utilized experimental data across multiple laboratories to model experimental variability and investigate subcellular phenotypic mechanisms in iPSC-CMs This framework links molecular mechanisms to cellular-level outputs by revealing unique subsets of model parameters linked to known iPSC-CM phenotypes ABSTRACT: There is a profound need to develop a strategy for predicting patient-to-patient vulnerability in the emergence of cardiac arrhythmia. A promising in vitro method to address patient-specific proclivity to cardiac disease utilizes induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). A major strength of this approach is that iPSC-CMs contain donor genetic information and therefore capture patient-specific genotype-phenotype relationships. A cited detriment of iPSC-CMs is the cell-to-cell variability observed in electrical activity. We postulated, however, that cell-to-cell variability may constitute a strength when appropriately utilized in a computational framework to build cell populations that can be employed to identify phenotypic mechanisms and pinpoint key sensitive parameters. Thus, we have exploited variation in experimental data across multiple laboratories to develop a computational framework for investigating subcellular phenotypic mechanisms. We have developed a whole-cell model of iPSC-CMs composed of simple model components comprising ion channel models with single exponential voltage-dependent gating variable rate constants, parameterized to fit experimental iPSC-CM data for all major ionic currents. By optimizing ionic current model parameters to multiple experimental datasets, we incorporate experimentally-observed variability in the ionic currents. The resulting population of cellular models predicts robust inter-subject variability in iPSC-CMs. This approach links molecular mechanisms to known cellular-level iPSC-CM phenotypes, as shown by comparing immature and mature subpopulations of models to analyse the contributing factors underlying each phenotype. In the future, the presented models can be readily expanded to include genetic mutations and pharmacological interventions for studying the mechanisms of rare events, such as arrhythmia triggers.


Evaluation of Pea and Soybean as Trap Crops for Managing Heterodera glycines.
Trap crops that stimulate nematode egg hatching but not reproduction have been reported as an effective means for managing certain nematodes. Studies were carried out at two field sites each year in 1998 and 1999 to evaluate the potential of trapping the soybean cyst nematode (Heterodera glycines) with soybean and pea in the corn year to manage the nematode in Minnesota. The trap crops were planted on the same day as corn at each site and later killed with the herbicide glyphosate. Nematode egg densities were determined at planting, 1 and 2 months after planting, and at harvest. Treatments included four seeding rates (0, 124,000, 247,000, and 494,000 seeds/ha) of resistant soybean as a trap crop and four kill dates (3, 4, 5, and 6 weeks after planting). No effects of the trap-crop and kill-date treatments on H. glycines population density, corn yield, and the followingyear soybean yield were observed at the two locations. In a second study, the experiment included four trap-crop comparisons (resistant soybean at 494,000 seeds/ha, susceptible soybean at 494,000 seeds/ha, pea at 1,482,000 seeds/ha, and no trap crop) and five kill dates (3, 4, 5, 6 weeks after planting, and no-kill). At the Waseca site, egg density at harvest was lower where resistant soybean was grown for 6 weeks and where pea was grown for 5 and 6 weeks compared with where no trap crop was grown. Maintaining pea plants for more than 5 weeks, however, reduced corn yield by 20% at the Waseca site. At the Lamberton site, egg density at harvest was lower where the susceptible soybean was grown for 5 weeks compared with where no trap crop was grown. Even with significant reduction of eggs in some treatments, use of soybean and pea as trap crops in the corn year was not an effective means for managing H. glycines.


Adenylate cyclase stimulation and intraocular pressure reduction by forskolin analogs.
Forskolin and 18 chemical analogs of forskolin were assayed for stimulation of adenylate cyclase activity in vitro and for their effects on intraocular pressure (IOP) in vivo. Adenylate cyclase activity was determined with a preparation of corpora striata from male Wistar rats and test drug concentrations of 0.3 to 300 microM. IOP effect was monitored after a single topical ocular application of a 1% suspension of the test drug to male New Zealand albino rabbits. Significant reductions in IOP occurred with compounds which had potent cyclase stimulatory properties. Most compounds with little cyclase stimulatory effect produced little or no decrease in IOP. A 1-substituted morpholino-acetoxy derivative, which may be a prodrug metabolized to active form by corneal esterases, showed little cyclase stimulation in vitro but produced significant reductions in IOP.


Breath analysis as a potential diagnostic tool for tuberculosis.
Cape Town, South Africa. We investigated the potential of breath analysis by gas chromatography-mass spectrometry (GC-MS) to discriminate between samples collected prospectively from patients with suspected tuberculosis (TB). Samples were obtained in a TB-endemic setting in South Africa, where 28% of culture-proven TB patients had Ziehl-Neelsen (ZN) negative sputum smear. A training set of breath samples from 50 sputum culture-proven TB patients and 50 culture-negative non-TB patients was analysed using GC-MS. We used support vector machine analysis for classification of the patient samples into TB and non-TB. A classification model with seven compounds had a sensitivity of 72%, a specificity of 86% and an accuracy of 79% compared with culture. The classification model was validated with breath samples from a different set of 21 TB and 50 non-TB patients from the same area, giving a sensitivity of 62%, a specificity of 84% and an accuracy of 77%. This study shows that GC-MS breath analysis is able to differentiate between TB and non-TB breath samples even among patients with a negative ZN sputum smear but a positive culture for Mycobacterium tuberculosis. We conclude that breath analysis by GC-MS merits further research.


Removal of natural hormones by nanofiltration membranes: measurement, modeling, and mechanisms.
The removal mechanisms of four natural steroid hormones-estradiol, estrone, testosterone, and progesterone-by nanofiltration (NF) membranes were investigated. Two nanofiltration membranes with quite different permeabilities and salt retention characteristics were utilized. To better understand hormone removal mechanisms, the membrane average pore size was determined from retention data of inert organic solutes of various molecular weights and a pore transport model that incorporates steric (size) exclusion and hindered convection and diffusion. Results indicate that, at the early stages of filtration, adsorption (or partitioning) of hormones to the membrane polymer is the dominant removal mechanism. Because the adsorptive capacity of the membrane is limited, the final retention stabilizes when the adsorption of hormones into the membrane polymer has reached equilibrium. At this later filtration stage, the overall hormone retention is lower than that expected based solely on the size exclusion mechanism. This behavior is attributed to partitioning and subsequent diffusion of hormone molecules in the membrane polymeric phase, which ultimately results in a lower retention. Hormone diffusion in the membrane polymeric matrix most likely depends on the size of the hormone molecule, hydrogen bonding of hormones to membrane functional groups, and hydrophobic interactions of the hormone with the membrane polymeric matrix.


Homodimerization of Ror2 tyrosine kinase receptor induces 14-3-3(beta) phosphorylation and promotes osteoblast differentiation and bone formation.
Ror2 receptor plays a key role in bone formation, but its signaling pathway is not completely understood. We demonstrate that Ror2 homodimerizes at the cell surface, and that dimerization can be induced by a bivalent antibody. Antibody-mediated dimerization causes receptor autophosphorylation and induces functional consequences of its signaling, including osteogenesis in mesenchymal stem cells and bone formation in organ culture. We further show that Ror2 associates with and phosphorylates 14-3-3beta scaffold protein. Endogenous Ror2 binds 14-3-3beta in U2OS osteosarcoma cells, and purified intracellular domain of Ror2 interacts with 14-3-3beta in vitro. 14-3-3beta Is tyrosine phosphorylated in U2OS cells, and this phosphorylation is inhibited by down-regulating Ror2 and enhanced by overexpressing the kinase. Purified Ror2 phosphorylates 14-3-3beta in vitro, confirming 14-3-3beta as the first identified Ror2 substrate. Down-regulating 14-3-3beta potentiates osteoblastogenesis in mesenchymal stem cells and increases bone formation in calvarial cultures, indicating that 14-3-3beta exerts a negative effect on osteogenesis. This raises a possibility that Ror2 induces osteogenic differentiation, at least in part, through a release of the 14-3-3beta-mediated inhibition. Our research forms a foundation for several new areas of investigation, including the molecular regulation of 14-3-3 by tyrosine phosphorylation and the role of this scaffold in osteogenesis.


Bacterial terpene cyclases.
Covering: up to 2015. This review summarises the accumulated knowledge about characterised bacterial terpene cyclases. The structures of identified products and of crystallised enzymes are included, and the obtained insights into enzyme mechanisms are discussed. After a summary of mono-, sesqui- and diterpene cyclases the special cases of the geosmin and 2-methylisoborneol synthases that are both particularly widespread in bacteria will be presented. A total number of 63 enzymes that have been characterised so far is presented, with 132 cited references.


Exposure to tetrachloroethylene in dry cleaning shops in the Nordic countries.
Tetrachloroethylene is the dominant solvent used in dry cleaning worldwide and many workers are potentially exposed. We report here on results of 1296 measurements of tetrachloroethylene undertaken in Nordic dry cleaning shops 1947-2001. We searched documents and files in the Nordic institutes of occupational health for air measurements of tetrachloroethylene. Repeated measurements from the same facility during a short time interval were registered only once using the time-weighted average. We registered also changes over time in occupational exposure limits (OELs) to tetrachloroethylene. Only scattered measurements were available from the early years, and the exposure level seemed fairly stable up until the mid 1970s. The median exposure level was 20 p.p.m. in 1976 and decreased to 3 p.p.m. in 2000. Exposure levels in the four Nordic countries followed similar trends. In the late 1960s, the OELs varied between the Nordic countries from 30 to 100 p.p.m. Sweden was first to lower the limit, but limits gradually converged over time. At present, Denmark, Finland, and Sweden use 10 p.p.m., while Norway uses 6 p.p.m. Over time, the average observed exposure level was lower than the OEL in all countries, but in Denmark and Sweden, up to one-third of measured exposures exceeded the OEL. Overall, the stationary measurements for maintenance work showed 36 p.p.m., while the personal measurements showed 7.5 p.p.m. for dry cleaners and 6.25 p.p.m. for shop assistants. The Nordic data illustrate that it is possible over time to control chemical exposures even in an industry consisting of many small and scattered work places.


Amoxicillin-clavulanate therapy increases childhood nasal colonization by methicillin-susceptible Staphylococcus aureus strains producing high levels of penicillinase.
We examined factors associated with penicillinase production by nasal carriage Staphylococcus aureus strains in 648 children aged 3 to 6 years attending 20 randomly sampled playschools. The children were prospectively monitored for drug use and medical events for 6 months and were then screened for S. aureus carriage. Isolates were tested for their susceptibility to penicillin G and methicillin, and penicillinase production by methicillin-susceptible, penicillin-resistant strains was quantified. S. aureus was isolated from 166 children (25.6%). Exposure to amoxicillin-clavulanate during the previous 3 months was associated with higher penicillinase production by penicillin-resistant, methicillin-susceptible strains (odds ratio, 3.6; P = 0.03). These results suggest that use of the amoxicillin-clavulanate combination could induce a herd selection process of S. aureus strains producing higher levels of penicillinase.


The current status of vital tooth whitening techniques.
Tooth whitening of vital teeth continues to have a major impact on the practice of dentistry. The growing public interest in having whiter, brighter teeth is clearly evident in the advertisements from toothpaste manufacturers on "whitening" formulations of their products and by the number of individuals seeking whitening procedures from their dentists. In addition, new over-the-counter whitening products continue to emerge in a marketplace that cannot seem to get teeth white enough, bright enough, fast enough. What new products and procedures have evolved over the past decade to whiten teeth? Are they better, safer, faster, and more effective now? Are dentists meeting public demand for whiter teeth and is this quest having a positive or negative impact on the practice of dentistry or the patient's dental health? I posed these questions to a group of experts on whitening procedures to get their opinions and recommendations.


Functional limitations and somatic diseases are independent predictors for incident depressive disorders in seniors: Findings from a nationwide longitudinal study.
Few nationwide comprehensive studies analyzed the factors leading to the onset of depression in correlation with medical disease and other related factors concerning geriatric depression. This study examined medical diseases with other factors which lead to depression among the elderly. This Taiwan-based longitudinal study examined a collection of 1467 seniors aged over 65. Subjects who fit this criteria were initially interviewed in 2003, and then four years later. Independent variables included baseline demographics, chronic medical illnesses, and the change of subjects' self-perceived health status, functional limitations including ADL, IADL and mobility limitation factors. The dependent variable was the symptoms of incident depression, as ascertained by the ten-item questionnaire during the later session. The logistic regression analyses were used to examine some of the predictors related to depressive disorders. The findings showed that heart conditions (adjusted OR=1.55, 95% CI: 1.12-2.15, p=0.008) and joint disorders (adjusted OR=1.51, 95% CI: 1.09-2.09, p=0.013), as well as functional limitations, particularly IADL (adjusted OR=1.81, 95% CI: 1.24-2.65, p=0.002) and ADL (adjusted OR=1.77, 95% CI: 1.27-2.47, p=0.001) were independently associated with the onset of depression among the elderly population. These findings indicated that when classifying symptoms of depression in geriatric patients with several underlying medical diseases, keen attention should be directed to the type of medical disorders and the functional deterioration in terms of daily activities and autonomic capabilities.


Sex reassignment at birth. Long-term review and clinical implications.
This article is a long-term follow-up to a classic case reported in pediatric, psychiatric, and sexological literature. The penis of an XY individual was accidentally ablated and he was subsequently raised as a female. Initially this individual was described as developing into a normally functioning female. The individual, however, was later found to reject this sex of rearing, switched at puberty to living as a male, and has successfully lived as such from that time to the present. The standard in instances of extensive penile damage to infants is to recommend rearing the male as a female. Subsequent cases should, however, be managed in light of this new evidence.


8q22-->qter duplication in a child with multiple congenital malformations: case report.
Congenital malformation syndromes are often caused by unbalanced chromosome translocations, which appear spontaneously or may be inherited from a healthy parent being the carrier of a balanced reciprocal translocation (rcp). Breakpoints, underlying chromosome fragment exchanges, may be located at any point of any chromosome and therefore, an infinite number of different translocations is possible. Special emphasis is placed both on the clinical characterization of every rare chromosomal aberration syndrome and on the determination of its breakpoints. Diagnosis of a 8q22-->qter duplication in a child with multiple congenital malformations. We determined the karyotypes of the five members of proband's family were established by using classical cytogenetic methods on whole blood obtained by venipuncture. We described a rare familial reciprocal translocation t(8; 14), observed in balanced form in mother and one healthy son, while being unbalanced in the son with congenital malformations. Balanced chromosome 8 aberration carriers should be aware of the procreation risks and need genetic counseling.


Water, sanitation, hygiene, and nutrition: successes, challenges, and implications for integration.
This study explores the integration of water, sanitation, and hygiene (WASH) and nutrition programming for improved child health outcomes and aims to identify barriers to and necessary steps for successful integration. Sixteen semi-structured in-depth interviews were conducted with key stakeholders from both the WASH and nutrition sectors, exploring barriers to integration and potential steps to more effectively integrate programs. Key barriers included insufficient and siloed funding, staff capacity and interest, knowledge of the two sectors, coordination, and limited evidence on the impact of integrated programs. To achieve more effective integration, respondents highlighted the need for more holistic strategies that consider both sectors, improved coordination, donor support and funding, a stronger evidence base for integration, and leadership at all levels. Organizations desiring to integrate programs can use these results to prepare for challenges and to know what conditions are necessary for successfully integrated programs. Donors should encourage integration and fund operational research to improve the efficiency of integration efforts. Knowledge among sectors should be shared and incentives should be designed to facilitate better coordination, especially where both sectors are working toward common goals.


Patient characteristics and prescription fill patterns for allergic rhinitis medications, with a focus on montelukast, in a commercially insured population.
Allergic rhinitis (AR), also known as hay fever, is caused by an overreaction of the immune system to airborne allergens. AR is a substantial cause of widespread morbidity, medical treatment costs, and reduced productivity at work and school. The goal of this research was to describe patient characteristics and prescription fill patterns for patients with AR and to determine those factors associated with the use of montelukast in a large population of commercially insured patients who sought medical treatment for AR. This was a retrospective cohort study using administrative claims data from a commercially available database. Patients, aged 4 to 64 years, with >or=3 years' continuous enrollment and >or=1 medical claim for AR between January 1, 2004, and December 31, 2006, were included. Patients with a concomitant asthma diagnosis were excluded. Patients' demographic and clinical characteristics, comorbidities, health care resource utilization, AR-related medication use, and AR-related physician office visits were assessed for 12 months before the first AR medication fill (index) in 2006. Stepwise logistic regression was used to identify factors predicting the initiation of montelukast therapy for the treatment of AR in 2006. The study population consisted of 75,140 children (mean [SD] age, 10.6 [4.0] years) and 226,236 adults (mean age, 43.8 [11.8] years). Slightly more than half (52.4%) of the pediatric population was male compared with 44.7% of the adult population. Fifty percent of patients had no pharmacy fills for an AR medication in 2006. Among patients with AR pharmacy fills (n = 150,751), 78.1% had a single index medication fill (montelukast represented 4.5%) and 21.9% were prescribed multiple index medications. Children with AR were more likely to fill a prescription for montelukast (n = 7513) if they were 4 to 11 years of age; male; diagnosed with cough/wheeze; and had 1 or 2 oral corticosteroid fills, >or=3 antibiotic fills, and AR-related physician office visits in the prior 12 months (all, P < 0.001). Prescription fills for montelukast among adult patients with AR were significantly (P < 0.001) associated with other respiratory/atopic conditions; prior fills for antihistamines, oral corticosteroids, or intranasal corticosteroids; and AR-related physician office visits in the prior 12 months. Children and adults with health plans based in the midwestern or southern region of the United States had greater odds of initiating montelukast than those with plans based in the western region (P < 0.001). Half of the patients identified with AR did not fill a prescription for an AR medication. Among those patients with AR-related prescription drug fills, most were prescribed a single index pharmacotherapy and did not receive additional AR medications within 30 days of the index date. The use of montelukast was limited and was more commonly prescribed to children and adults with AR whose condition was not controlled with other AR medications.


Community readiness and momentum: identifying and including community-driven variables in a mixed-method rural palliative care service siting model.
Health service administrators make decisions regarding how to best use limited resources to have the most significant impact. Service siting models are tools that can help in this capacity. Here we build on our own mixed-method service siting model focused on identifying rural Canadian communities most in need of and ready for palliative care service enhancement through incorporating new community-driven insights. We conducted 40 semi-structured interviews with formal and informal palliative care providers from four purposefully selected rural communities across Canada. Communities were selected by running our siting model, which incorporated GIS methods, and then identifying locations suitable as qualitative case studies. Participants were identified using multiple recruitment methods. Interviews were transcribed verbatim and the transcripts were reviewed to identify emerging themes and were coded accordingly. Thematic analysis then ensued. We previously introduced the inclusion of a 'community readiness' arm in the siting model. This arm is based on five community-driven indicators of palliative care service enhancement readiness and need. The findings from the current analysis underscore the importance of this arm of the model. However, the data also revealed the need to subjectively assess the presence or absence of community awareness and momentum indicators. The interviews point to factors such as educational tools, volunteers, and local acknowledgement of palliative care priorities as reflecting the presence of community awareness and factors such as new employment and volunteer positions, new care spaces, and new projects and programs as reflecting momentum. The diversity of factors found to illustrate these indicators between our pilot study and current national study demonstrate the need for those using our service siting model to look for contextually-relevant signs of their presence. Although the science behind siting model development is established, few researchers have developed such models in an open way (e.g., documenting every stage of model development, engaging with community members). This mixed-method study has addressed this notable knowledge gap. While we have focused on rural palliative care in Canada, the process by which we have developed and refined our siting model is transferrable and can be applied to address other siting problems.


Interventions to improve neurodevelopmental outcomes of children born moderate to late preterm: a systematic review protocol.
Introduction: Prematurity (birth before 37 +0 weeks' gestation) is associated with wide-ranging neurodevelopmental impairment. Prognosis among moderate to late (32 +0 to <37 +0 weeks' gestation) preterm infants (MLPT) is better compared to their counterparts born very preterm (<32 +0 weeks' gestation). However the risk of developmental impairment among MLPT, who make up about 84% of all preterm infants, is 2-3 times higher when compared to infants born at term. Early interventions have aimed to improve outcomes in preterm infants generally, but there are limited data on their need and effect in MLPT specifically. Prioritising research, long-term follow-up and early interventions targeted at ameliorating the impact of preterm birth among MLPT is required. Objectives: To conduct a systematic review of the type of early childhood interventions (from birth until 4 years of age) offered to MLPT children and to evaluate their impact on neurodevelopmental outcomes (cognitive, neurobehavioural and motor) as assessed in these children during childhood (until 18 years of age). Methods and analysis: A systematic literature search in Web of Science, Medline Ovid, PsycINFO, CINAHL and EMBASE will be conducted. Data on MLPT children receiving developmental interventions until the age of 4 years will be evaluated. Interventions may involve parents or primary caregivers. Primary outcomes are cognitive, neurobehavioural and motor development as measured from birth until the age of 18 years. The Cochrane Risk of Bias Assessment Tool will be used to evaluate the methodological quality of randomised controlled trials (RCTs) included in the review and will be graded as low, high or unclear risk of bias. The quality of non-RCTs will be evaluated with the Newcastle-Ottawa Scale. The quality of evidence for each outcome will be evaluated using the Grading of Recommendations Assessment, Development and Evaluation Approach. Publication and reporting bias will be assessed using Egger's test and funnel plots respectively.


Studies on the mechanism underlying the influence of alanine infusion on glucose dynamics in the dog.
These experiments have been designed to study the influence of alanine infusion of glucose dynamics in the dog and to further elucidate the role of pancreatic hormones in the interaction of alanine with glucose homeostasis. The primed constant infusion of glucose-2-t was used in order to quantitate the rates of glucose production by the liver (Ra) and glucose utilization (Rd). In a first group of experiments, the intravenous infusion of alanine at the rate of 2 mg./kg./min. produced a moderate enhancement of plasma insulin (IRI), while pancreatic glucagon (IRG) increased more consistently. This different pattern of IRI and IRG response caused the insulin/glucagon molar ratio to decline progressibely throughout the experiment. Both rates of glucose turnover increased significantly during alanine infusion. Since Ra rose more rapidly thanRd did initially, hyperglycemia developed. Later, glucose production slowly decreased and, in spite of the sustained hyperglucagonemia, reached levels very close to the baseline in the second part of the experiment. A significant direct correlation between Ra and IRG was found, while the changes in Ra correlated inversely with those in I/G molar ratio. In a second group of experiments, alanine was infused at the same dose together with 0.4 microng./kg./min. of cyclic somatostatin. In the first part of the infusion, IRG fell more than IRI did, so that I/G ratio increased. Later, IRI levels maintained at low values while IRG returned slowly to the baseline and consequently I/G ratio significantly decreased. Glucose production fell rapidly soon after the beginning of the infusion, and therefore hypoglycemia developed. Later, Ra increased progressively to levels above baseline and plasma glucose returned to the preinfusion levels. As in the the first group of experiments, a significant direct correlation between Ra and IRG and an inverse correlation between the changes in Ra and I/G ratio were observed. These experiments demonstrate that alanine infusion produces an acceleration of glucose turnover and that a clear interrelationship between the release of glucose by the liver and the mobilization of pancreatic hormones exists. Finally, the experiments with somatostatin indicate that hyperglucagonemia is one of the mechanisms underlying the stimulatory effect of alanine on glucose production.


Effect of intraoral aging on the setting status of resin composite and glass ionomer orthodontic adhesives.
The aim of this study was to assess the effect of intraoral aging on the setting status of a resin composite and a glass ionomer adhesive, relative to control specimens stored in water. Metallic brackets were bonded with resin composite orthodontic adhesive (Transbond XT; 3M Unitek, Monrovia, Calif) or a glass ionomer cement (Fuji I; GC, Tokyo, Japan) to recently extracted premolars and kept in water for 6 months. The same materials were also bonded to the premolars of orthodontic patients. After 6 months, the teeth were carefully extracted, with the brackets intact on their buccal surfaces. All teeth were embedded in epoxy resin and sectioned buccolingually. Fourier transform infrared microscopy and Raman microscopy were used for the estimation of the degree of cure in the composite and the salt yield in the glass ionomer adhesives. The control samples of the composite showed significantly lower degrees of cure than did the retrieved specimens (52.40% ± 3.21% vs 57.62% ± 1.32% by Fourier transform infrared microscopy, and 61.40% ± 2.61% vs 67.40% ± 3.44% by Raman microscopy). Raman microscopy significantly overestimated the degree of cure and failed to provide reliable information for the salt yield in the glass ionomer cement. Fourier transform infrared microscopy showed increased, but no statistically significant difference in, aluminum-carboxylate salts in the retrieved specimens. Enhanced oxidation of residual carbon-carbon bonds in the composite and slightly increased dissolution of the weaker calcium-salt phase in the glass ionomer cement were the main differences in the intraorally aged specimens in comparison with the specimens stored in water.


[Detection, isolation and characterization of Shiga toxin-producing Escherichia coli (STEC) in fresh ground beef from butcher shops in Concepción, Tucumán Province].
Shiga toxin-producing Escherichia coli is an emerging foodborne pathogen. There are many STEC serotypes associated with human diseases, being the O157:H7 serotype the most prevalent. Ground beef is the main transmission vehicle. In Concepción city, Tucumán Province, between September and December 2004, two hemolytic uremic syndrome (HUS) cases were diagnosed. The main objective of this work was to detect, isolate and characterize STEC O157 and non-O157 strains in fresh ground beef. Between September and December 2004, 53 fresh ground beef samples were collected from butcher shops in Concepción city. The USDA-FSIS (2002) methodology was used for detection, isolation and characterization of STEC O157:H7. Two PCR techniques for E. coli non-O157 detection and a previous intra-laboratory validated methodology for the isolation and characterization of these strains were used. The stx2 gen was identified in seven samples and the rfbO157 gene also in four of them. However, only one E. coli O157:H7 strain, biotype C, carrying the eae, stx2 and ehxA genes, was isolated. The present study shows the importance of implementing techniques for the detection of this emerging pathogen in meat samples.


A Population-Based Descriptive Atlas of Invasive Pneumococcal Strains Recovered Within the U.S. During 2015-2016.
Invasive pneumococcal disease (IPD) has greatly decreased since implementation in the U.S. of the 7 valent conjugate vaccine (PCV7) in 2000 and 13 valent conjugate vaccine (PCV13) in 2010. We used whole genome sequencing (WGS) to predict phenotypic traits (serotypes, antimicrobial phenotypes, and pilus determinants) and determine multilocus genotypes from 5334 isolates (~90% of cases) recovered during 2015-2016 through Active Bacterial Core surveillance. We identified 44 serotypes; 26 accounted for 98% of the isolates. PCV13 serotypes (inclusive of serotype 6C) accounted for 1503 (28.2%) isolates, with serotype 3 most common (657/5334, 12.3%), while serotypes 1 and 5 were undetected. Of 305 isolates from children <5 yrs, 60 (19.7%) were of PCV13 serotypes 19A, 19F, 3, 6B, and 23F (58/60 were 19A, 19F, or 3). We quantitated MLST-based lineages first detected during the post-PCV era (since 2002) that potentially arose through serotype-switching. The 7 predominant emergent post-PCV strain complexes included 23B/CC338, 15BC/CC3280, 19A/CC244, 4/CC439, 15A/CC156, 35B/CC156, and 15BC/CC156. These strains accounted for 332 isolates (6.2% of total) and were more frequently observed in children <5 yrs (17.7%; 54/305). Fifty-seven categories of recently emerged (in the post PCV7 period) putative serotype-switch variants were identified, accounting for 402 isolates. Many of these putative switch variants represented newly emerged resistant strains. Penicillin-nonsusceptibility (MICs > 0.12 μg/ml) was found among 22.4% (1193/5334) isolates, with higher penicillin MICs (2-8 μg/ml) found in 8.0% (425/5334) of isolates that were primarily (372/425, 87.5%) serotypes 35B and 19A. Most (792/1193, 66.4%) penicillin-nonsusceptible isolates were macrolide-resistant, 410 (34.4%) of which were erm gene positive and clindamycin-resistant. The proportion of macrolide-resistant isolates increased with increasing penicillin MICs; even isolates with reduced penicillin susceptibility (MIC = 0.06 μg/ml) were much more likely to be macrolide-resistant than basally penicillin-susceptible isolates (MIC < 0.03 μg/ml). The contribution of recombination to strain diversification was assessed through quantitating 35B/CC558-specific bioinformatic pipeline features among non-CC558 CCs and determining the sizes of gene replacements. Although IPD has decreased greatly and stabilized in the post-PCV13 era, the species continually generates recombinants that adapt to selective pressures exerted by vaccines and antimicrobials. These data serve as a baseline for monitoring future changes within each invasive serotype.


Endoleaks after endovascular abdominal aortic aneurysm repair: detection with dual-energy dual-source CT.
To assess the diagnostic performance of dual-energy dual-source computed tomography (CT) in the detection of endoleaks after endovascular abdominal aortic aneurysm (AAA) repair. This study was local ethics board approved, and written informed consent was obtained from all patients. One hundred eighteen patients (21 women, 97 men; mean age, 74 years +/- 8 [standard deviation]) underwent follow-up dual-energy dual-source CT during the nonenhanced, arterial, and delayed phases after AAA repair. Delayed phase CT images were acquired in the dual-energy mode for reconstruction of virtual nonenhanced images. Two blinded and independent readers evaluated the data for the presence or absence of endoleaks during three reading sessions: Standard nonenhanced, arterial phase, and delayed phase images were read during session A; virtual nonenhanced, arterial phase, and delayed phase images, during session B; and virtual nonenhanced and delayed phase images, during session C. Sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) were calculated, with the session A image data set as the reference standard. Radiation dose parameters were estimated. Reading session A revealed that 52 (44%) of 118 patients had endoleaks. Overall sensitivity, specificity, NPV, and PPV for CT endoleak detection during sessions B and C were identical: 100%, 97%, 100%, and 96%, respectively. The accuracy of the session B and session C readings was not significantly different from that of the session A reading (P = .50). The effective radiation dose in the image acquisition protocol involving one dual-energy scan was significantly (P < .001) lower than the effective doses in the protocols involving standard triple-phase scanning (mean difference, 61%) and standard nonenhanced and delayed phase scanning (mean difference, 41%). Compared with standard protocols, one dual-energy dual-source CT scan performed during the delayed phase with reconstruction of virtual nonenhanced images enables detection of endoleaks after endovascular AAA repair with high accuracy and a considerably lower radiation dose.


Novel genes for potential ligand-binding proteins in subregions of the olfactory mucosa.
Odorant detection is specifically mediated via receptor neurons in the olfactory mucosa but is a complex process involving a number of different cell types producing proteins of differing function. We have used the technique of subtractive hybridization cDNA cloning to identify novel genes expressed exclusively in the olfactory mucosa which may play a role in olfaction. Ten distinct groups of cDNA clones were identified which corresponded to mRNA transcripts highly expressed in rat olfactory mucosa but undetectable in thymus, kidney, lung, brain, spleen and liver. Some of these clones identify substructures in the mucosal tissue for which no other probes are currently available. Others identify novel mRNA species in the Bowman's glands. The predicted proteins for three of these clones are homologous to proteins which bind to either lipopolysaccharides (RYA3 and RY2G5) or to polychlorinated biphenyls (RYD5). In addition, while RYA3 and RY2G5 are highly homologous, they appear to be expressed in different parts of the mucosal tissue. The sequence homologies and subanatomical location of expression suggest that these proteins might interact with odorants before or after specific recognition by odorant receptors. Therefore, the olfactory mucosa may possess diverse, functionally-distinct odorant-binding proteins which recognize and bind separate classes of odorants.


A note on the musculature of the proximal part of the pelvic limb of the ostrich (Struthio camelus).
Ostriches (Struthio camelus) (n=90) were dissected and the musculature of part of the proximal pelvic limb examined. Graphical representation of these muscles are presented. It is evident that the anatomy of the hip and thigh of the ostrich is typically avian, but with only the M. iliofemoralis present. The M. iliotrochantericus medius is not found in ostriches, but both the M. iliofemoralis externus and M. iliofemoralis internus, as well as both parts of the M. flexor cruris lateralis and the M. ambiens are presnet in ostriches. The ambiens muscle, responsible for the automatic grasping action in birds, originates from the preacetabular lateral surface of the ilium. The insertion of this muscle is, as in other birds, via a long tendon on the head of the M. flexor digitorum magnus. A pectineus muscle is present in ostriches. The Mm. femorotibiales consists of 4 muscles, inclusive of the M. femorotibialis accessorius.


Evaluation and Behavior of Spanish Bodybuilders: Doping and Sports Supplements.
The use of doping agents has these days become a public health problem, as it also affects young and non-competitive amateurs in different sports. To prepare for competition, bodybuilders perform aggressive dietary protocols, so, bodybuilders frequently consume nutritional supplements (NS) and banned substances in large dosages. Thus, the aim of this study is to analyze the prevalence of banned substances consumption and NS intake in competitive level bodybuilders. A total of 48 bodybuilders (44 males and 4 females) completed a validated online questionnaire on NS consumption. The quantitative data was presented as a mean (M) ± standard deviation (SD), as well as having minimum and maximum values. The categorical variables were expressed using frequencies and percentages. 83.3% of the participants declared that they had consumed or would consume banned substances, the most consumed being anabolic steroids (72.9%). One hundred percent of those sampled use NS. Whey protein (96%), branched-chain amino acids (BCAA) (94%), creatine (85%) and vitamin complexes (83%) were the most consumed, however, there is a low consumption of certain NS which could also increase athletic performance.


The quality of trabecular bone evaluated with micro-computed tomography, FEA and mechanical testing.
Most standard methods to predict bone quality are merely based on apparent density measurements. However, apparent density alone does neither explain all variation of the mechanical properties nor does it account for the structural anisotropy of trabecular bone. Thus, apparent density alone might not be sufficient to accurately predict the quality of bone. This study investigates if a new approach based on microstructural computer models can provide additional and relevant information on bone quality. 58 human trabecular bone samples from the femoral head were measured with a 3-D micro-Computed Tomography (micro-CT) system providing a voxel representation of the bone microarchitecture with a resolution of 28 microns. Based on such representations, the orthotropic stiffness matrices and the principal directions were computed for 5 mm cubes with microstructural Finite Element Analysis (FEA). For a subset of six samples the moduli were then validated with tri-axial mechanical compression tests. The results show that on average 15% of the variation of the elastic properties are not explained by bone volume fraction. Differences of elastic properties between samples with the same bone volume fraction range up to 53%. The variation of the degree of anisotropy is unrelated to that of the bone volume fraction. Finally, the direction-dependent stiffness of the trabecular bone differs by a factor of four, indicating that one single (isotropic) modulus as predicted from apparent density measurements might not be adequat. It is concluded that micro-CT-based FEA provides new and additional information about anisotropy and mechanical properties in a direct and non-destructive way, and thus will be important in the future for advanced failure risk prediction. An extension to patient examinations using high-resolution CT or MRI techniques is envisaged.


Assessment of pesticide safety knowledge and practices in Vietnam: A cross-sectional study of smallholder farmers in the Mekong Delta.
Over the past three decades, the Vietnamese Mekong Delta has experienced a significant increase in agricultural productivity, partly achieved through increased agrochemical use. To abate negative effects on human and environmental health, several national programs were launched to enhance safer pesticide use. This study aimed to assess the patterns and relationships of official sustainable agriculture educational programs, pesticide safety knowledge, and practices of smallholder farmers in the Mekong Delta. A cross-sectional survey was conducted with 400 smallholder farmers from three communes in Thoi Lai district (Can Tho province) from March to May 2020. Twenty-four questions on pesticide safety knowledge and practices were used to identify traits using latent class analysis. Adjusted generalized linear regression was used to assess determinants of pesticide safety knowledge and estimate associations of pesticide safety knowledge with pesticide practices. 96.2% of participants have used at least one WHO class II pesticide during the past year while the use of specific personal protective equipment was limited mainly due to unavailability (37.0%) or discomfort (83.0%). High education (Odds Ratio (OR), 95% Confidence Interval; 3.84, 1.70-9.45), exposure to official educational programs (1.87, 1.13-3.12), peer-to-peer knowledge exchange (3.58, 2.18-6.00), and learning from governmental extension services (2.31, 1.14-4.98) were positively associated with increased pesticide safety knowledge. Compared to poor practices, pesticide safety knowledge was increasingly positively associated with intermediate (1.65, 1.02-2.66) and good pesticide practices (8.96, 2.58-31.12). These findings highlight the importance of school education and educational programs, access to PPE, and addressing discomforts of PPE to improve the protection of farmers from pesticide exposures. Simultaneously, pesticide market authorization processes should be reconsidered to promote the authorization of less toxic products. Further in-depth studies on the nature of pesticides used, nonuse of personal protective equipment, and effectiveness of educational programs will further define leverage points for safer pesticide use.


A nonlinear analysis of the cerebrospinal fluid system and intracranial pressure dynamics.
A mathematical model of the cerebrospinal fluid (CSF) system was developed to help clarify the kinetics of the intracranial pressure (ICP). A general equation predicting the time course of pressure was derived in terms of four parameters: the intracranial compliance, dural sinus pressure, resistance to absorption, and CSF formation. These parameters were measured in the adult cat, and the equation was tested by comparing experimental and calculated values of the time course of pressure in response to volume changes. The theoretical and experimental results were in close agreement, and the role of each parameter in governing the dynamic equilibrium of the ICP was determined. From this analysis, dynamic tests were developed for rapid measurement of CSF formation, absorption resistance, and the bulk intracranial compliance. These techniques are applicable to clinical settings, providing data that are useful in characterizing the physiological mechanisms responsible for raised ICP and assessing changes induced by therapy.


Combination effects of AHR agonists and Wnt/β-catenin modulators in zebrafish embryos: Implications for physiological and toxicological AHR functions.
Wnt/β-catenin signaling regulates essential biological functions and acts in developmental toxicity of some chemicals. The aryl hydrocarbon receptor (AHR) is well-known to mediate developmental toxicity of persistent dioxin-like compounds (DLCs). Recent studies indicate a crosstalk between β-catenin and the AHR in some tissues. However the nature of this crosstalk in embryos is poorly known. We observed that zebrafish embryos exposed to the β-catenin inhibitor XAV939 display effects phenocopying those of the dioxin-like 3,3',4,4',5-pentachlorobiphenyl (PCB126). This led us to investigate the AHR interaction with β-catenin during development and ask whether developmental toxicity of DLCs involves antagonism of β-catenin signaling. We examined phenotypes and transcriptional responses in zebrafish embryos exposed to XAV939 or to a β-catenin activator, 1-azakenpaullone, alone or with AHR agonists, either PCB126 or 6-formylindolo[3,2-b]carbazole (FICZ). Alone 1-azakenpaullone and XAV939 both were embryo-toxic, and we found that in the presence of FICZ, the toxicity of 1-azakenpaullone decreased while the toxicity of XAV939 increased. This rescue of 1-azakenpaullone effects occurred in the time window of Ahr2-mediated toxicity and was reversed by morpholino-oligonucleotide knockdown of Ahr2. Regarding PCB126, addition of either 1-azakenpaullone or XAV939 led to lower mortality than with PCB126 alone but surviving embryos showed severe edemas. 1-Azakenpaullone induced transcription of β-catenin-associated genes, while PCB126 and FICZ blocked this induction. The data indicate a stage-dependent antagonism of β-catenin by Ahr2 in zebrafish embryos. We propose that the AHR has a physiological role in regulating β-catenin during development, and that this is one point of intersection linking toxicological and physiological AHR-governed processes.


Cell-type-specific trans-activation of herpes simplex virus thymidine kinase promoter by the human T-cell leukemia virus type I Tax protein.
The human T-cell leukemia virus type I Tax protein (HTLV-I Tax) is known as a trans-activating factor for a variety of genes, including those of cytokines. Here, we show that Tax is capable of activating the herpes simplex virus thymidine kinase (HSV-TK) promoter in certain mammalian cell lines. In murine NIH 3T3 fibroblasts and human HeLa cells, trans-activation by Tax was remarkably strong, whereas in human chondrocytic HCS-2/8 and monkey kidney Cos-7 cells, the responsiveness of the TK promoter to Tax was poor. Deletion analysis revealed that one of the two previously described Sp1 sites is required for the Tax responsiveness, whereas the CTF binding site is not. The results suggest possible interactions between the oncogenic Tax protein and the viral TK in coinfected cells in vivo. Care should be taken in the context of HTLV-I research, as the HSV-TK promoter has been widely used in molecular biology and gene therapeutics.


Impact of high drinking water nitrate levels on the endogenous formation of apparent N-nitroso compounds in combination with meat intake in healthy volunteers.
Nitrate is converted to nitrite in the human body and subsequently can react with amines and amides in the gastrointestinal tract to form N-nitroso compounds (NOCs), which are known to be carcinogenic in animals. Humans can be exposed to nitrate via consumption of drinking water and diet, especially green leafy vegetables and cured meat. The contribution of nitrate from drinking water in combination with meat intake has not been investigated thoroughly. Therefore, in the present pilot study, we examined the effect of nitrate from drinking water, and its interaction with the consumption of white and processed red meat, on the endogenous formation of NOCs, taking into account the intake of vitamin C, a nitrosation inhibitor. Twenty healthy subjects were randomly assigned to two groups consuming either 3.75 g/kg body weight (maximum 300 g per day) processed red meat or unprocessed white meat per day for two weeks. Drinking water nitrate levels were kept low during the first week (< 1.5 mg/L), whereas in week 2, nitrate levels in drinking water were adjusted to the acceptable daily intake level of 3.7 mg/kg bodyweight. At baseline, after 1 and 2 weeks, faeces and 24 h urine samples were collected for analyses of nitrate, apparent total N-nitroso compounds (ATNC), compliance markers, and genotoxic potential in human colonic Caco-2 cells. Urinary nitrate excretion was significantly increased during the high drinking water nitrate period for both meat types. Furthermore, levels of compliance markers for meat intake were significantly increased in urine from subjects consuming processed red meat (i.e. 1-Methylhistidine levels), or unprocessed white meat (i.e. 3-Methylhistidine). ATNC levels significantly increased during the high drinking water nitrate period, which was more pronounced in the processed red meat group. Genotoxicity in Caco-2 cells exposed to faecal water resulted in increased genotoxicity after the interventions, but results were only significant in the low drinking water nitrate period in subjects consuming processed red meat. Furthermore, a positive correlation was found between the ratio of nitrate/vitamin C intake (including drinking water) and the level of ATNC in faecal water of subjects in the processed red meat group, but this was not statistically significant. Drinking water nitrate significantly contributed to the endogenous formation of NOC, independent of the meat type consumed. This implies that drinking water nitrate levels should be taken into account when evaluating the effect of meat consumption on endogenous formation of NOC. Dutch Trialregister: 29707 . Registered 19th of October 2018. Retrospectively registered.


High sputum total adiponectin is associated with low odds for asthma.
Adipose tissue produces adiponectin, an anti-inflammatory protein. High systemic total adiponectin is associated with a low risk for incident asthma but the association with lung adiponectin is not known. Our objective was to evaluate the association between sputum total adiponectin and asthma. This case-control study included 44 cases with objectively-confirmed asthma and an equal number of body mass index (BMI) and sex-matched controls. Serum and sputum adiponectin were estimated by ELISA and Western Blot technique, respectively. While Fisher's exact test, t-test and Spearman's correlations were used for univariate analyses, Spearman and regression analyses were performed for multivariable analyses. While high-molecular-weight adiponectin was the dominant isoform in serum, medium-molecular-weight isoform was dominant in sputum. Sputum total adiponectin was not correlated with serum adiponectin or BMI. Sputum total adiponectin was lower among asthmatics than controls (p = 0.03), although individual sputum isoforms were not similarly associated. High sputum total adiponectin was associated with lower odds for asthma (OR 0.33, 95% C.I. 0.12, 0.91), even after adjustment for systemic adiposity measures including serum adiponectin. High sputum total adiponectin predicted lower odds for asthma, even after adjustment for serum adiponectin. Although not studied, it is possible that pharmacological modulation of sputum adiponectin may suggest new ways to prevent and/or treat asthma.


Improved results for postcardiotomy cardiogenic shock with the use of implantable left ventricular assist devices.
Over the past decade, the use of mechanical circulatory support in patients with postcardiotomy cardiogenic shock has resulted in hospital discharge rates of 25% to 40%. In an attempt to improve patient survival, we initiated a program of early insertion of an implantable Thermocardiosystems Incorporated Heartmate left ventricular assist device in patients who have circulatory failure after having undergone high-risk cardiac operations. Between April 1993 and February 1997, 12 patients underwent insertion of an implantable left ventricular assist device for postcardiotomy cardiogenic shock after coronary artery bypass grafting. Indications for insertion included postoperative cardiogenic shock (7 patients), postoperative cardiac arrest (3 patients), and failure to wean from cardiopulmonary bypass (2 patients). The median time to device insertion was 3.5 days. The mean duration of left ventricular assist device support was 103 +/- 19 days (range, 2 to 225 days). Nine of 11 patients (82%) survived to undergo either transplantation (8 patients) or explantation (1 patient), with successful hospital discharge of all 9 patients. The major complication was device-related infection (42%). A single thromboembolism occurred in a patient with an infection. Long-term outcome after postcardiotomy cardiogenic shock is improved substantially with the use of an implantable left ventricular assist device early in the postoperative course. Access to such a device is an important consideration before undertaking a high-risk cardiac operation, and early implantation of the device is a critical factor in ensuring long-term survival.