Application of molecularly imprinted polymers in the anti-doping field: sample purification and compound analysis.
The problem posed by anti-doping requirements is one of the great analytical challenges; multiple compound detection at low ng ml-1 levels from complex samples, with requirements for exceptional confidence in results. This review surveys the design, synthesis and application of molecularly imprinted polymers (MIPs) in this field, focusing on the templating of androgenous anabolic steroids (AASs), as the most commonly abused substances, but also other WADA prohibited substances. Commentary on the application of these materials in detection, clean-up and sensing is offered, alongside views on the future of imprinting in this field.


[Gelastic epilepsy and precocious puberty in hamartoma of the hypothalamus].
Four cases of hypothalamic hamartoma leading to gelastic epilepsy, precocious puberty and behavioural disorders are reported. Cerebral neuroradiologic examinations revealed a tumor-like mass attached to the hypothalamus in the region of the mamillary bodies in all cases. Precocious puberty developed in the two girls at 4 and 13 months but in neither of the two boys, who both suffered behaviour disturbances in the form of aggressive outbursts. A total resection of the tumors of both boys led to histologic confirmation of hamartoma. One boy was free of seizures upon follow-up, whereas seizure frequency in the other boy was reduced, while his aggressivity increased. The cases are discussed in context of current therapeutic conceptions of gelastic epilepsy and central precocious puberty.


A Comparison Study on Traditional Mixtures of Herbal Teas Used in Eastern Mediterranean Area.
Multipurpose herbal teas with numerous ingredients, in which flowers are the main component, are common in the traditional medicine and pharmacy of Greece and the Eastern Mediterranean countries. In this study, we combine ethnobotany and ethnopharmacology field work techniques and botany and pharmacognosy laboratory methods for the study of traditional herbal mixtures with flowers, we identify their botanical ingredients and record the local medicinal uses of these mixtures, in Greece, Lebanon, Syria, Iran and Turkey. These, and their industrial versions, are analyzed, using morphological and multivariate analysis techniques in order to determine marker species, relevant patterns of combination and local styles. The medicinal properties attributed to the different flowers are discussed in relation with their role in the mixtures. These blends are consumed for their relaxing, digestive, and anti-infective properties. These mixtures are not consumed as a treatment when one is sick but rather to avoid getting sick, as a preventive measure. The formulations can reach forty ingredients (sarantha in Greek, arbain in Arabic language of Palestine), usually entire or coarsely chopped in the more traditional formulations, leading to extreme variability of individual doses. We ask what biological signification this randomness can have. To give an answer requires new and more comprehensive pharmacological approaches. The flowers of Rosaceae, Asteraceae, Lamiaceae, Malvaceae and Fabaceae species characterize these mixtures in which other materials (roots, leaves, and fruits) and other species are present as well. Flowers of some species, particularly of Fabaceae, are exclusively used in mixtures, and their use in monospecific herbal teas is not yet recorded. We draw attention on the urgent need in exhaustively recording in Greece and the Near East, the formulation and use of traditional herbal mixtures and their numerous local variants. To consider these mixtures and the contribution of flowers (most mixtures receive the general name of tea of flowers) merits further extensive study.


An electrically evoked slow potential of the frog's retina. I. Properties of response.
1. Brief pulses of electrical current passed through the frog's eye cup from sclera to vitreous (inward current) evoked large, vitreous positive slow potentials of retinal origin. Brief pulses of outward current evoke no response. 2. This electrically evoked retinal response (EERG) increased sharply in amplitude as the strength of the stimulus was increased. 3. Strength-duration curves showed that the stimulating pulse was integrated with a time constant ranging from 14 to 36 ms. The time constant was a function of the amplitude chosen as a criterion. 4. The EERG was abolished by chemical agents which abolish the PII component of the ERG (KCl, NH3, aspartate). 5. The waveform of the EERG returned to the base line with exactly the same time course as the PII component of the ERG, both when the PII was evoked by flashes of light and when it was evoked by flashes of darkness. 6. These results can be explained in terms of the hypothesis that electrical stimuli act on the synaptic terminals of the photoreceptor cells.


Adhesion networks of cnidarians: a postgenomic view.
Cell-extracellular matrix (ECM) and cell-cell adhesion systems are fundamental to the multicellularity of metazoans. Members of phylum Cnidaria were classified historically by their radial symmetry as an outgroup to bilaterian animals. Experimental study of Hydra and jellyfish has fascinated zoologists for many years. Laboratory studies, based on dissection, biochemical isolations, or perturbations of the living organism, have identified the ECM layer of cnidarians (mesoglea) and its components as important determinants of stem cell properties, cell migration and differentiation, tissue morphogenesis, repair, and regeneration. Studies of the ultrastructure and functions of intercellular gap and septate junctions identified parallel roles for these structures in intercellular communication and morphogenesis. More recently, the sequenced genomes of sea anemone Nematostella vectensis, Hydra magnipapillata, and coral Acropora digitifera have opened up a new frame of reference for analyzing the cell-ECM and cell-cell adhesion molecules of cnidarians and examining their conservation with bilaterians. This chapter integrates a review of literature on the structure and functions of cell-ECM and cell-cell adhesion systems in cnidarians with current analyses of genome-encoded repertoires of adhesion molecules. The postgenomic perspective provides a fresh view on fundamental similarities between cnidarian and bilaterian animals and is impelling wider adoption of species from phylum Cnidaria as model organisms.


Fatty acid metabolism in patients with PPARgamma mutations.
PPARG mutations may cause insulin resistance and dyslipidemia, but little is known about the mechanisms of the abnormalities of lipid metabolism. We hypothesized that in PPARG mutations, abnormal adipose tissue triglyceride storage causes insulin resistance. Whole-body and adipose tissue-specific metabolic phenotyping through arteriovenous blood sampling was made before and after a mixed meal including 13C-palmitic acid. Studies were performed in a 32-yr-old male with partial lipodystrophy and type 2 diabetes, heterozygous for the PPARG P467L mutation and in an apparently phenotypically normal 32-yr-old male heterozygous for the PPARG n.AAA553T mutation. Comparator groups were age- and sex-matched healthy participants (n=10) and type 2 diabetes sex-matched participants (n=6). The P467L patient had elevated unmodulated fasting and postprandial plasma nonesterified fatty acid (NEFA) concentrations, despite a low adipose tissue NEFA output. Instead, NEFA appeared to originate directly from triglyceride-rich lipoproteins: 13C-palmitic acid accumulated rapidly in the NEFA fraction, as a sign of impaired fatty acid trapping in tissues. In contrast to the Pparg haploinsufficient mouse, the patient with n.AAA553T mutation did not exhibit paradoxically insulin sensitive and showed a mostly normal metabolic pattern. The lipodystrophic PPARG P467L phenotype include excessive and uncontrolled generation of NEFA directly from triglyceride-rich lipoproteins, explaining high systemic NEFA concentrations, whereas the human PPARG haploinsufficiency is metabolically almost normal.


Political Economy of Reform under US Federalism: Adopting Single-Payer Health Coverage in New York State.
The US remains the only high-income country that lacks a universal health financing system and instead relies on a fragmented system with the largest segment of the population receiving health insurance through private, voluntary employer-sponsored health insurance plans. While not "universal" in the sense of being mandatory and tax-financed, through a series of reforms, the US has managed to provide some form of health insurance coverage to 90% of the population. Yet, the high cost of this system, the insufficient coverage afforded to many, and continued concerns about equity have led to calls for a national health insurance program that can reduce costs across the board while providing high-quality coverage for all. Given the policy gridlock at the national level, the states have often sought to achieve universal health financing on their own, but these bills have met with little success so far. Why has the ideal of states as "laboratories of democracy" failed to produce policy change towards national health insurance? This article examines the prospects for the New York Health Act, a single-payer bill that would create a universal health financing plan for all New York State residents. Applying the Political Economy of Health Financing Framework, we analyze the politics of health reform in New York State and identify strategies to overcome opposition to this policy proposal. We find that while a clear political opportunity is in place, the prospects for adoption remain low given the power of symbolic politics and institutional inertia on the reform process.


Anti-HTLV-1 immunity combined with proviral load as predictive biomarkers for adult T-cell leukemia-lymphoma.
Human T-cell leukemia virus type 1 (HTLV-1) establishes chronic infection in humans and induces a T-cell malignancy called adult T-cell leukemia-lymphoma (ATL) and several inflammatory diseases such as HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Persistent HTLV-1 infection is established under the pressure of host immunity, and therefore the immune response against HTLV-1 is thought to reflect the status of the disease it causes. Indeed, it is known that cellular immunity against viral antigens is suppressed in ATL patients compared to HAM/TSP patients. In this study, we show that profiling the humoral immunity to several HTLV-1 antigens, such as Gag, Env, and Tax, and measuring proviral load are useful tools for classifying disease status and predicting disease development. Using targeted sequencing, we found that several carriers whom this profiling method predicted to be at high risk for developing ATL indeed harbored driver mutations of ATL. The clonality of HTLV-1-infected cells in those carriers was still polyclonal; it is consistent with an early stage of leukemogenesis. Furthermore, this study revealed significance of anti-Gag proteins to predict high risk group in HTLV-1 carriers. Consistent with this finding, anti-Gag cytotoxic T lymphocytes (CTLs) were increased in patients who received hematopoietic stem cell transplantation and achieved remission state, indicating the significance of anti-Gag CTLs for disease control. Our findings suggest that our strategy that combines anti-HTLV-1 antibodies and proviral load may be useful for prediction of the development of HTLV-1-associated diseases.


Surgical Preparation for Articular Cartilage Regeneration in the Osteoarthritic Knee Joint.
Autologous chondrocyte implantation (ACI) is a treatment option even in early osteoarthritis (OA). Surgical preparation for ACI should avoid penetration of the subchondral bone plate to prevent hemorrhage, fibrin clot formation, and subsequent activation of the inflammatory response. Current surgical procedures with ring curettes preserve the integrity of the subchondral bone plate, even in patients with OA. Subchondral femoral bone plates ( n = 40) of OA knees undergoing total knee arthroplasty were prepared in vivo using standard, non-brute-force debridement for ACI. To approach regular wear/early OA, only cartilage with maximally grade 3A International Cartilage Repair Society score was prepared. Effects were analyzed by light microscopy. In 87.5% of the specimens (35/40), standard debridement did not violate the tide mark, except for occasional minor openings with a smooth edge (diameter approximately 20 µm). In contrast, 5/40 samples (12.5%) showed one large area with a missing bone plate and an open bone marrow space. Twenty-eight specimens (70%) showed at least remnants of uncalcified cartilage. On the basis of size/fine structure, the occasional minor openings are likely due to increased vascular penetration through the tide mark in the pathologically altered bone-cartilage interface in OA. The consequences of limited hemorrhage through minor openings or selected large defects following in vivo debridement are still unknown. Thus, standard debridement appears suitable for cartilage regeneration even in OA defects.


Influence of incubation temperature on the behavior of triethylamine-extractable glyphosate (N-phosphonomethylglycine) in four soils.
The behavior of glyphosate, extracted from four soils using aqueous triethylamine, was investigated at two temperatures. For each soil, and at both temperatures, there was a marked loss in the amount of extractable glyphosate immediately after addition of the herbicide to soil. This rapid loss of glyphosate was ascribed to adsorption of the herbicide into a nonextractable form. For three of the four soils used when incubated at 25 degrees C, the rates of loss of extractable glyphosate were similar to previously measured rates of degradation of this herbicide in these soils. However, loss of extractable glyphosate from the Culgoa clay loam was due not only to substrate degradation but also to slow sorption of glyphosate into the nonextractable form in this soil over the experimental period. For the Rutherglen and Walpeup soils, when incubated at 10 degrees C, the rates of loss of extractable glyphosate were half of the previously measured rate of degradation of this herbicide in these soils. However, there was no measured loss of extractable glyphosate from the Wimmera clay. As previous work has shown glyphosate to decompose readily in these soils at this temperature, these findings suggest that desorption of glyphosate may occur at a rate greater than degradation at this temperature and, hence, that temperature may play a pivotal role in sorption processes. Investigations with these soils when sterilized by gamma-irradiation showed that for the Walpeup, Wimmera, and Rutherglen soils, sorption was complete soon after the addition of the herbicide; however, for the Culgoa soil, further adsorption occurred over the entire experimental period.


Human activin betaA gene. Identification of novel 5' exon, functional promoter, and enhancers.
On the basis of cDNA cloning, primer extension, and transfection experiments, we identified a novel 5' exon of the human activin betaA subunit gene, and found its enhancer and promoter regions as well as multiple transcription start sites. A series of deletion and mutation analyses of the enhancer sequences defined the 45-base pair core region (DR-1 core) containing two short elements with similarity to AP-1 (12-O-tetradecanoylphorbol-13-acetate response element; TRE) and CREB/ATF (cyclic AMP response element; CRE) binding sites, both of which were necessary for full enhancer activity. Gel shift and antibody supershift assays using DR-1 core region revealed the formation of two specific DNA-protein complexes, one of which could be partially dissociated by a competing oligonucleotide containing a single copy of the consensus TRE, but the other of which contained neither CREB/ATF nor AP-1 as major components. Although 12-O-tetradecanoylphorbol-13-acetate and cAMP induced the activin enhancer/promoter-driven CAT activity, such drug induction was obscured when either the TRE- or CRE-like elements were mutated in the native promoter context. Our results demonstrate that the promoter and enhancer regions identified here are essential for maintaining the efficient promoter activity of the human activin betaA subunit gene.


Chronic fatigue syndrome: a review for clinicians.
Syndromes characterized by persistent fatigue, musculoskeletal pain, sleep disturbance, and subjective cognitive impairment have been common problems in clinical practice for decades. The chronic fatigue syndrome case definition was created to standardize the patient population in research studies and to foster a systematic and comprehensive approach to the attempt to define the etiology and pathophysiology of these syndromes. The pathogenesis of chronic fatigue syndrome remains unknown, though it does appear to be associated with subtle neuroendocrine and immunologic abnormalities. Treatment of chronic fatigue syndrome is empirical. Significant palliation is often possible, though treatment success requires skillful practice of the art of medicine.


Epigenetic and inflammatory markers in older adults with cancer: A Young International Society of Geriatric Oncology narrative review.
The number of adults aged ≥ 65 years with cancer is rapidly increasing. Older adults with cancer are susceptible to treatment-related acute and chronic adverse events, resulting in loss of independence, reduction in physical function, and decreased quality of life. Nevertheless, evidence-based interventions to prevent or treat acute and chronic adverse events in older adults with cancer are limited. Several promising blood-based biomarkers related to inflammation and epigenetic modifications are available to identify older adults with cancer who are at increased risk of accelerated aging and physical, functional, and cognitive impairments caused by the cancer and its treatment. Inflammatory changes and epigenetic modifications can be reversible and targeted by lifestyle changes and interventions. Here we discuss ways in which changes in inflammatory and epigenetic pathways influence the aging process and how these pathways can be targeted by interventions aimed at reducing inflammation and aging-associated biological markers. As the number of older adults with cancer entering survivorship continues to increase, it is becoming progressively more important to understand ways in which the benefit from treatment can be enhanced while reducing the effects of accelerated aging.


Next generation pharmaceutical impactor (a new impactor for pharmaceutical inhaler testing). Part II: Archival calibration.
A new seven-stage cascade impactor, the Next Generation Pharmaceutical Impactor (NGI), has been developed for the pharmaceutical industry. A calibration following "good laboratory practice (GLP)" procedures has been performed on a specific archival NGI, deemed to be representative of all NGIs. Thus, this impactor had nozzle dimensions for each stage manufactured close to the middle of the tolerance band for the design specification, and therefore the average nozzle diameter was equal to the nominal value for that stage. An essential feature of the NGI is that it is designed to operate at any flow rate between 30 and 100 L/min. Thus, the calibration was made at inlet flow rates of 30, 60 and 100 L/min representing the lower bound, mid-region and upper bound of the specified range of operation for the impactor. The calibration data were then used to develop equations that predict the particle cut size for all components of the impactor at any flow rate from 30 to 100 L/min.


Measurement of personal competence of mentally retarded individuals.
The development and validation of the Personal Competency Scale, a 20-item rating instrument designed to assess competence of mentally retarded adults, was described. This broad bandwidth measure provides scores on three factorially derived domain scales, which are consistent with the rationale underlying the instrument's development, and a total score. The three major dimensions of personal competence delineated were adaptive, cognitive, and affective competence. Reliability estimates in the form of interrater, test-retest, and internal consistency were high, ranging from .80 to .94 for the subscales and .90 to .97 for the total scale. Correlations between the Personal Competency Scale and criterion variables, including IQ and Adaptive Behavior Scale scores, ranged from .61 to .75. Multiple regression and canonical correlations of .85 and .87, respectively, also strongly support the validity of this new measure.


Expanding walls and shrinking beaches: loss of natural coastline in Okinawa Island, Japan.
Okinawa is the largest and most populated island of the Ryukyu Archipelago in southern Japan and is renowned for its natural resources and beauty. Similar as to what has been happening in the rest of the country, Okinawa Island has been affected by an increasing amount of development and construction work. The trend has been particularly acute after reversion to Japanese sovereignty in 1972, following 27 years of post-war American administration. A coastline once characterized by extended sandy beaches surrounded by coral reefs now includes tracts delimited by seawalls, revetments, and other human-made hardening structures. Additionally, part of coastal Okinawa Island was obtained by land-filling shallow ocean areas (land reclamation). Nevertheless, the current extension of the artificial coastline, as well as the level of fragmentation of the natural coastline are unclear, due to the lack of both published studies and easily accessible and updated datasets. The aims of this research were to quantify the extension of coastline alterations in Okinawa Island, including the amount of land-filling performed over the last 41 years, and to describe the coastlines that have been altered the most as well as those that are still relatively pristine. The analyses were performed using a reference map of Okinawa Island based on GIS vector data extracted from the OpenStreetMap (OSM) coastline dataset (average node distance for Okinawa Island = 24 m), in addition to satellite and aerial photography from multiple providers. We measured 431.8 km of altered coastline, equal to about 63% of the total length of coastline in Okinawa Island. Habitat fragmentation is also an issue as the remaining natural coastline was broken into 239 distinct tracts (mean length = 1.05 km). Finally, 21.03 km2 of the island's surface were of land reclaimed over the last 41 years. The west coast has been altered the most, while the east coast is in relatively more natural conditions, particularly the northern part, which has the largest amount of uninterrupted natural coastline. Given the importance of the ecosystem services that coastal and marine ecosystems provide to local populations of subtropical islands, including significant economic income from tourism, conservation of remaining natural coastlines should be given high priority.


Circulatory miRNA-155, miRNA-21 target PTEN expression and activity as a factor in breast cancer development.
Breast cancer is a complex disease with multiple factors involved in its pathophysiological development. genetic mutations of BRCA1, BRCA2 and p53 are among the most well-studied factors. The role of other genetic factors like altered expression profiles, SNPs in the regulatory regions of different genes or epigenetic factors like promoter methylation and histone modifications are also well studied but no solid understanding is available on distinct key players triggering malignancy in breast cancer, (Phosphatase and tensin homolog) PTEN is known to be a crucial tumor suppressor as it has been reported to be missing or abnormally expressed in many cancer cells. Here in this were studied how PTEN is expressed in malignant and benign cancer cells by investigating its expression profile and cellular location using Immuno-fluorescence microscopy. At the same time, quantitative studies of the circulatory mi-RNAs related to the downregulation of PTEN, namely mir-21 and mir-155 have studied also. Sixty biopsy samples, forty were diagnosed to be malignant and twenty were benign. It has been found that PTEN is normally expressed in benign samples and its normally localized in the cell membrane, while in malignant samples the expression level of PTEN is lower or absent and it is translocated to the cytoplasm. Interestingly the quantitative expression of circulatory mir-21 and mir-155 in the blood plasma of the corresponding patients showed a related pattern with higher expression in malignant samples, therefore can it's clear that PTEN is in the cross-talk of genetics and epigenetic regulation in regard of the development of malignant breast cancer. At the same time, this study confirms the importance of circulatory miRNAs as a biomarker for early breast cancer detection.


Facility-Level Excess Mortality of VHA Patients With Mental Health or Substance Use Disorder Diagnoses.
Individuals with mental or substance use disorders have higher mortality rates than people in the general population. How excess mortality varies across health care facilities is unknown. The authors sought to investigate facility-level mortality rates among Veterans Health Administration (VHA) patients who had received diagnoses of mental or substance use disorders. An electronic medical records-based retrospective cohort study was conducted, encompassing 8,812,373 unique users of 139 VHA facilities from 2011 to 2016. Covariates included age, sex, and past-year diagnoses of serious mental illness, posttraumatic stress disorder, major depressive disorder, other mental health conditions, or substance use disorders. The outcome was all-cause mortality per comprehensive Veterans Affairs/Department of Defense searches of the National Death Index. Proportional hazards regression was used to calculate overall and facility-specific hazard ratios (HRs) for each diagnosis group, adjusted for age, sex, and comorbid medical conditions. Overall, all-cause mortality was statistically significantly elevated among VHA users with mental health diagnoses (HR=1.21, 95% confidence interval=1.20-1.22). HRs varied across facilities consistently over time. At the VHA facility level, diagnostic groups were significantly correlated with the degree of excess mortality. Results were similar in sensitivity analyses that excluded deaths from suicide or drug or alcohol overdose. VHA users with mental or substance use disorder diagnoses had elevated mortality rates. Correlation in excess mortality across two periods indicated that facility differences in excess mortality were persistent and therefore potentially associated with facility- and community-level factors, which may help inform quality improvement efforts to reduce mortality rates.


The Bone Dysplasia Ontology: integrating genotype and phenotype information in the skeletal dysplasia domain.
Skeletal dysplasias are a rare and heterogeneous group of genetic disorders affecting skeletal development. Patients with skeletal dysplasias suffer from many complex medical issues including degenerative joint disease and neurological complications. Because the data and expertise associated with this field is both sparse and disparate, significant benefits will potentially accrue from the availability of an ontology that provides a shared conceptualisation of the domain knowledge and enables data integration, cross-referencing and advanced reasoning across the relevant but distributed data sources. We introduce the design considerations and implementation details of the Bone Dysplasia Ontology. We also describe the different components of the ontology, including a comprehensive and formal representation of the skeletal dysplasia domain as well as the related genotypes and phenotypes. We then briefly describe SKELETOME, a community-driven knowledge curation platform that is underpinned by the Bone Dysplasia Ontology. SKELETOME enables domain experts to use, refine and extend and apply the ontology without any prior ontology engineering experience--to advance the body of knowledge in the skeletal dysplasia field. The Bone Dysplasia Ontology represents the most comprehensive structured knowledge source for the skeletal dysplasias domain. It provides the means for integrating and annotating clinical and research data, not only at the generic domain knowledge level, but also at the level of individual patient case studies. It enables links between individual cases and publicly available genotype and phenotype resources based on a community-driven curation process that ensures a shared conceptualisation of the domain knowledge and its continuous incremental evolution.


Site of osteodensitometry in perimenopausal women: correlation and limits of agreement between anatomic regions.
Because the bone mineral density (BMD) in different anatomic regions is heterogenous the number of women who fulfill the World Health Organization definition of osteopenia or osteoporosis increases with the number of regions examined. The purpose of this study was to investigate the agreement between measurements of the spine, femur, forearm, and whole body following menopause. Two thousand and five healthy, perimenopausal women, mean age 50.6 years, were studied using Hologic QDR-1000/W and QDR-2000 densitometers. Though the BMD of different anatomic regions were correlated (r = 0.40-0.77, p < 0.01), the variability in each patient regarding T and Z scores between regions was considerable. For example, despite a high correlation (r = 0.67, p < 0.01) and no systematic difference between the T scores for total femoral and lumbar BMD, the limits of agreement (mean difference +/- 2 SD) for the comparison were -1.89 to 1.87. Femoral neck T scores were 0.5 SD lower than those of the other regions, confirming reports that the young adult reference for this measurement is disproportionally high. The prevalence of osteoporosis was 1.2% when femur total BMD was considered alone and 5.9% when lumbar and ultradistal forearm results were included. However, as many as 7.9% showed osteoporosis of the femoral neck when the Hologic T score was used, compared with 0.7% using National Health and Nutrition Examination Survey (NHANES) values. The choice of anatomic region and availability of appropriate young adult reference data has considerable impact on the apparent prevalence of osteoporosis. Given the heterogeneity between regions, a combination of spinal and femoral densitometry should be used in diagnosing osteoporosis, though this increases the prevalence of osteoporosis by 50% or more in perimenopausal women.


Discovery of sexual dimorphisms in metabolic and genetic biomarkers.
Metabolomic profiling and the integration of whole-genome genetic association data has proven to be a powerful tool to comprehensively explore gene regulatory networks and to investigate the effects of genetic variation at the molecular level. Serum metabolite concentrations allow a direct readout of biological processes, and association of specific metabolomic signatures with complex diseases such as Alzheimer's disease and cardiovascular and metabolic disorders has been shown. There are well-known correlations between sex and the incidence, prevalence, age of onset, symptoms, and severity of a disease, as well as the reaction to drugs. However, most of the studies published so far did not consider the role of sexual dimorphism and did not analyse their data stratified by gender. This study investigated sex-specific differences of serum metabolite concentrations and their underlying genetic determination. For discovery and replication we used more than 3,300 independent individuals from KORA F3 and F4 with metabolite measurements of 131 metabolites, including amino acids, phosphatidylcholines, sphingomyelins, acylcarnitines, and C6-sugars. A linear regression approach revealed significant concentration differences between males and females for 102 out of 131 metabolites (p-values<3.8×10(-4); Bonferroni-corrected threshold). Sex-specific genome-wide association studies (GWAS) showed genome-wide significant differences in beta-estimates for SNPs in the CPS1 locus (carbamoyl-phosphate synthase 1, significance level: p<3.8×10(-10); Bonferroni-corrected threshold) for glycine. We showed that the metabolite profiles of males and females are significantly different and, furthermore, that specific genetic variants in metabolism-related genes depict sexual dimorphism. Our study provides new important insights into sex-specific differences of cell regulatory processes and underscores that studies should consider sex-specific effects in design and interpretation.


Image-aided estimate of tumor burden in Hodgkin's disease: evidence of its primary prognostic importance.
To explore a more direct method for evaluating tumor burden (TB) in Hodgkin's disease (HD) and to verify its prognostic importance. The volume of TB at diagnosis was directly and retrospectively measured in 121 HD patients through images of the lesions recorded by computed tomographic (CT) scan of the thorax, abdomen, and pelvis for all deep sites of involvement and many superficial ones, and by ultrasonography (US) for the remaining superficial lesions. The TB, which was obtained from the sum of the volumes of all the lesions measured on CT scans and US and normalized to body-surface area (relative TB [rTB]), showed a median value of 102.6 cm(3)/m(2) (range, 2.2 to 582.8). At multivariate analysis for prognostic value, rTB was the parameter that statistically correlated best with time to treatment failure (P = 2.2 x 10(-6)), followed by erythrocyte sedimentation rate (ESR) (P =.0003), and serum fibrinogen (P =.0112). The prognostic discrimination allowed by rTB alone proved to be clearly superior to that obtained with the score of the International Prognostic Factor Project. The rTB was found to be correlated with many clinical staging parameters (bulky disease, number of involved lymph node regions, serum lactate dehydrogenase, ESR, hemoglobin, Karnofsky index), but its predictability from these variables was low (R(2) =.668). Relative TB is emerging as a strong prognostic factor in HD, more powerful than and largely independent of those hitherto known and used. Further studies are needed to confirm these results and exploit their clinical value, particularly the relationship among rTB, drug doses, and response.


Biogenic amines and their metabolites are differentially affected in the Mecp2-deficient mouse brain.
Rett syndrome (RTT, MIM #312750) is a severe neurological disorder caused by mutations in the X-linked methyl-CpG binding protein 2 (MECP2) gene. Female patients are affected with an incidence of 1/15000 live births and develop normally from birth to 6-18 months of age before the onset of deficits in autonomic, cognitive, motor functions (stereotypic hand movements, impaired locomotion) and autistic features. Studies on Mecp2 mouse models, and specifically null mice, revealed morphological and functional alterations of neurons. Several functions that are regulated by bioaminergic nuclei or peripheral ganglia are impaired in the absence of Mecp2. Using high performance liquid chromatography, combined with electrochemical detection (HPLC/EC) we found that Mecp2(-/y) mice exhibit an alteration of DA metabolism in the ponto-bulbar region at 5 weeks followed by a more global alteration of monoamines when the disease progresses (8 weeks). Hypothalamic measurements suggest biphasic disturbances of norepinephrine and serotonin at pathology onset (5 weeks) that were found stabilized later on (8 weeks). Interestingly, the postnatal nigrostriatal dopaminergic deficit identified previously does not parallel the reduction of the other neurotransmitters investigated. Finally, dosage in cortical samples do not suggest modification in the monoaminergic content respectively at 5 and 8 weeks of age. We have identified that the level of catecholamines and serotonin is differentially affected in Mecp2(-/y) brain areas in a time-dependent fashion.


Ibuprofen-loaded ethylcellulose/polystyrene microspheres: an approach to get prolonged drug release with reduced burst effect and low ethylcellulose content.
The aim of this study was to develop ethylcellulose microspheres for prolonged drug delivery with reduced burst effect. Ethylcellulose microspheres loaded with ibuprofen were prepared with and without polystyrene, which was used to retard drug release from ethylcellulose microspheres. Ibuprofen-loaded ethylcellulose microspheres with a polystyrene content of 0-25% were prepared by the solvent evaporation technique and characterized by drug loading, infrared spectroscopy, differential scanning calorimetry and scanning electron microscopy. The in vitro release studies were performed to study the influence of polystyrene on ibuprofen release from ethylcellulose microspheres. The microspheres showed 28-46% of drug loading and 80-92% of entrapment, depending on polymer/drug ratio. The infrared spectrum and thermogram showed stable character of ibuprofen in the microspheres and revealed an absence of drug polymer interaction. The prepared microspheres were spherical in shape and had a size range of 0.1-4 microm. Ethylcellulose/polystyrene microspheres showed prolonged drug release and less burst effect when compared to microspheres prepared with ethylcellulose alone. Microspheres prepared with an ethylcellulose/polystyrene ratio of 80:20 gave a required release pattern for oral drug delivery. The presence of polystyrene above this ratio gave release over 24 h. To find out the mechanism of drug release from ethylcellulose/polystyrene microspheres, the data obtained from in vitro release were fitted in various kinetic models. High correlation was obtained in Higuchi and Korsmeyer-Peppas models. The drug release from ethylcellulose/polystyrene microspheres was found to be diffusion controlled.


Lateralized repetition priming for familiar faces: Evidence for asymmetric interhemispheric cooperation.
Repetition priming refers to facilitated recognition of stimuli that have been seen previously. Although a great deal of work has examined the properties of repetition priming for familiar faces, little has examined the neuroanatomical basis of the effect. Two experiments are presented in this paper that combine the repetition priming paradigm with a divided visual field methodology to examine lateralized recognition of familiar faces. In the first experiment participants were presented with prime faces unilaterally to each visual field and target faces foveally. A significant priming effect was found for prime faces presented to the right hemisphere, but not for prime faces presented to the left hemisphere. In Experiment 2, prime and target faces were presented unilaterally, either to the same visual field or to the opposite visual field (i.e., either within hemisphere or across hemispheres). A significant priming effect was found for the within right hemisphere condition, but not for the within left hemisphere condition, replicating the findings of the first experiment. Priming was also found in both of the across hemispheres conditions, suggesting that interhemispheric cooperation occurs to aid recognition. Taken in combination these experiments provide two main findings. First, an asymmetric repetition priming effect was found, possibly as a result of asymmetric levels of activation following recognition of a prime face, with greater priming occurring within the right hemisphere. Second, there is evidence for asymmetric interhemispheric cooperation with transfer of information from the right hemisphere to the left hemisphere to facilitate recognition.


Electrical fields enhance growth of cancer spheroids by reactive oxygen species and intracellular Ca2+.
A single electrical field pulse (500 V/m) with a duration of 60 s increased tumor outgrowth over a postpulse period of 24 h. RNA staining with acridine orange showed a rise in RNA content in pulsed spheroids, indicating stimulation of cell cycle activity. The electropulse induced an intracellular Ca2+ concentration ([Ca2+]i) transient that started approximately 40 s after the onset of the electrical field. Neither the presence of extracellular Ni2+ (0.5 mM) nor the absence of extracellular Ca2+ impeded the [Ca2+]i rise. It was, however, totally blocked by thapsigargin (1 microM), indicating that the initial Ca2+ response is due to Ca2+ release from intracellular stores. The [Ca2+]i transient was paralleled by an increase in reactive oxygen species (ROS), as revealed using 2',7'-dichlorofluorescein diacetate as an indicator. The radical scavengers N-acetyl-L-cysteine (NAC)(20 mM) and dehydroascorbate (5 mM) inhibited both ROS production and the [Ca2+]i transient during electrical field treatment. The mitogenic activation was dependent on the rise in [Ca2+]i because inhibition of Ca2+ release during electrical field treatment by addition of either thapsigargin or NAC to the incubation medium abolished the observed effect. We conclude that a single, direct current electrical field pulse induces production of ROS, which in turn mediate Ca2+ release from intracellular stores and activate cell cycle activity in multicellular spheroids.


Structure-activity relationships among alpha-(N)-heterocyclic acyl thiosemicarbazones and related compounds as inhibitors of herpes simplex virus type 1-specified ribonucleoside diphosphate reductase.
2-Acetylpyridine thiosemicarbazone, a potent antiviral drug, and 13 analogues were examined as inhibitors of partially purified herpes simplex virus type 1-specified ribonucleoside diphosphate reductase. N4,N4-Azacycloheptane derivatives were more active than their N4-unsubstituted analogues. Selenosemicarbazones were similar in potency to their thiosemicarbazone congeners, whereas a related semicarbazone was much less active. Maximum inhibition was observed when an ethylidene side-chain was present in the compounds. No discernible trend in potency was observed when the pyridine moiety was replaced by quinoline or isoquinoline. Thiosemicarbazide derivatives were less potent than their unsaturated thiosemicarbazone analogues. Inhibitory potencies increased at longer incubation times consistent with the hypothesis that thiosemicarbazones inactivate the enzyme in a time-dependent manner.


Metabolomic profiling of anionic metabolites by capillary electrophoresis mass spectrometry.
We describe a sheath flow capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) method in the negative mode using a platinum electrospray ionization (ESI) spray needle, which allows the comprehensive analysis of anionic metabolites. The material of the spray needle had significant effect on the measurement of anions. A stainless steel spray needle was oxidized and corroded at the anodic electrode due to electrolysis. The precipitation of iron oxides (rust) plugged the capillary outlet, resulting in shortened capillary lifetime. Many anionic metabolites also formed complexes with the iron oxides or migrating nickel ion, which was also generated by electrolysis and moved toward the cathode (the capillary inlet). The metal-anion complex formation significantly reduced detection sensitivity of the anionic compounds. The use of a platinum ESI needle prevented both oxidation of the metals and needle corrosion. Sensitivity using the platinum needle increased from several- to 63-fold, with the largest improvements for anions exhibiting high metal chelating properties such as carboxylic acids, nucleotides, and coenzyme A compounds. The detection limits for most anions were between 0.03 and 0.87 micromol/L (0.8 and 24 fmol) at a signal-to-noise ratio of 3. This method is quantitative, sensitive, and robust, and its utility was demonstrated by the analysis of the metabolites in the central metabolic pathways extracted from mouse liver.


[Extracorporeal shock waves induce ventral-periosteal new bone formation out of the focus zone--results of an in-vivo animal trial].
It is known that high-energy extracorporeal shock waves, focussed to the intact distal rabbit femur, induce periosteal new bone formation within the focus zone. This study was done to investigate whether there is additionally new bone formation outside of the focus zone, i. e., in the proximal femur, 10 days after shock wave application to the distal rabbit femur. 18 adult Chinchilla-Bastard rabbits were randomly divided into three groups (A, B, C; each n = 6). One distal femur of each animal was focussed for shock wave application with different positive energy flux densities (A 1.2 mJ/mm (2), B 0.9 mJ/mm (2), C sham treatment). Only the proximal femur outside the shock wave focus was investigated. Here periosteal new bone formation was evaluated by fluorescent labelling. The maximum thickness was measured by means of computer-assisted image analysis. Maximum ventral-periosteal new bone formation outside the shock wave focus was significantly increased in group A compared to group B (p = 0.007) or group C (p = 0.001). In contrast, the maximum ventral-periosteal new bone formation outside the shock wave focus demonstrated no statistically significant difference in group B compared to group C (p = 1.0). Depending from the amount of positive energy flux density applied to distal rabbit femur a significantly increased ventral-periosteal new bone formation results in the proximal rabbit femur outside the shock wave focus.


Ionic and synaptic mechanisms of seizure generation and epileptogenesis.
The biophysical mechanisms underlying epileptogenesis and the generation of seizures remain to be better understood. Among many factors triggering epileptogenesis are traumatic brain injury breaking normal synaptic homeostasis and genetic mutations disrupting ionic concentration homeostasis. Impairments in these mechanisms, as seen in various brain diseases, may push the brain network to a pathological state characterized by increased susceptibility to unprovoked seizures. Here, we review recent computational studies exploring the roles of ionic concentration dynamics in the generation, maintenance, and termination of seizures. We further discuss how ionic and synaptic homeostatic mechanisms may give rise to conditions which prime brain networks to exhibit recurrent spontaneous seizures and epilepsy.


Expression of the human interferon-gamma cDNA in yeast.
Several plasmids which direct the expression of human interferon-gamma (IFN-gamma) cDNA in the yeast Saccharomyces cerevisiae under the transcriptional control of the 3-phosphoglycerate kinase (PGK) promoter have been constructed. Up to 25 x 10(6) units per liter culture of biologically active IFN-gamma are synthesized in yeast transformed with these expression plasmids. The amount of IFN-gamma mRNA in yeast transformed with the IFN-gamma expression plasmids is much lower than the amount of PGK mRNA present when the PGK gene and promoter are inserted on a similar plasmid. The major and three minor sites of transcription initiation have been localized. The major starting point of the IFN-gamma mRNA is at 4 basepairs upstream of the PGK mRNA start. All transcription starts map at an ApG as does PGK mRNA. The yeast IFN-gamma mRNA terminates in a specific segment of the 3' untranslated region of the IFN-gamma cDNA. The polyadenylation site at about 182-184 nucleotides 3' of the IFN-gamma stop codon is preceded by a sequence which is very similar to a sequence proposed to be involved in transcription termination and polyadenylation (49).


Deployment morbidity among search-and-rescue dogs used after the September 11, 2001, terrorist attacks.
To determine characteristics, variables associated with deployment morbidity, and injuries and illnesses of search-and-rescue dogs associated with the Sept 11, 2001, terrorist attacks. Historical cohort study. 96 dogs. Data collected included previous medical or surgical history, physical attributes of dogs, type and number of years of training, site of deployment, shift and hours worked, and number of days deployed. Combined morbidity was defined as 1 or more abnormalities of body systems, including traumatic injuries. Handlers of 96 of the 212 dogs responded to the surveys. Fifty-nine dogs were deployed by the Federal Emergency Management Agency, 10 by police forces, and 27 as members of other search-and-rescue teams. Sixty-five dogs (incidence rate, 17 events/1,000 dog search hours) had combined morbidity during deployment. System-specific morbidity rates included gastrointestinal tract signs (5 events/1,000 dog search hours), cuts and abrasions mostly on the feet (5 events/1,000 dog search hours), fatigue (6 events/1,000 dog search hours), change in appetite (6 events/1,000 dogs search hours), dehydration (5 events/1,000 dog search hours), respiratory tract problems (2 events/1,000 dog search hours), heat exhaustion (2 events/1,000 dog search hours), and orthopedic or back problems (2 events/1,000 dog search hours). Dogs deployed to the World Trade Center were 6.6 times more likely to have combined morbidity, compared with dogs at the Pentagon. Injury and illnesses occurred in most dogs and affected several organ systems, but all were minor.


Lenalidomide plus dexamethasone is more effective than dexamethasone alone in patients with relapsed or refractory multiple myeloma regardless of prior thalidomide exposure.
This analysis assessed the efficacy and safety of lenalidomide + dexamethasone in patients with relapsed or refractory multiple myeloma (MM) previously treated with thalidomide. Of 704 patients, 39% were thalidomide exposed. Thalidomide-exposed patients had more prior lines of therapy and longer duration of myeloma than thalidomide-naive patients. Lenalidomide + dexamethasone led to higher overall response rate (ORR), longer time to progression (TTP), and progression-free survival (PFS) versus placebo + dexamethasone despite prior thalidomide exposure. Among lenalidomide + dexamethasone-treated patients, ORR was higher in thalidomide-naive versus thalidomide-exposed patients (P = .04), with longer median TTP (P = .04) and PFS (P = .02). Likewise for dexamethasone alone-treated patients (P = .03 for ORR, P = .03 for TTP, P = .06 for PFS). Prior thalidomide did not affect survival in lenalidomide + dexamethasone-treated patients (36.1 vs 33.3 months, P > .05). Thalidomide-naive and thalidomide-exposed patients had similar toxicities. Lenalidomide + dexamethasone resulted in higher rates of venous thromboembolism, myelosuppression, and infections versus placebo + dexamethasone, independent of prior thalidomide exposure. Lenalido-mide + dexamethasone was superior to placebo + dexamethasone, independent of prior thalidomide exposure. Although prior thalidomide may have contributed to inferior TTP and PFS compared with thalidomide-naive patients, these parameters remained superior compared with placebo + dexamethasone; similar benefits compared with placebo + dexamethasone were not evident for thalidomide-exposed patients in terms of overall survival. Studies were registered at http://www.clinicaltrials.gov under NCT00056160 and NCT00424047.


Radiofrequency ablation in primary non-small cell lung cancer: What a radiologist needs to know.
Lung cancer continues to be one of the leading causes of death worldwide. In advanced cases of lung cancer, a multimodality approach is often applied, however with poor local control rates. In early non-small cell lung cancer (NSCLC), surgery is the standard of care. Only 15-30% of patients are eligible for surgical resection. Improvements in imaging and treatment delivery systems have provided new tools to better target these tumors. Stereotactic body radiation therapy (SBRT) has evolved as the next best option. The role of radiofrequency ablation (RFA) is also growing. Currently, it is a third-line option in stage 1 NSCLC, when SBRT cannot be performed. More recent studies have demonstrated usefulness in recurrent tumors and some authors have also suggested combination of RFA with other modalities in larger tumors. Following the National Lung Screening Trial (NLST), screening by low-dose computed tomography (CT) has demonstrated high rates of early-stage lung cancer detection in high-risk populations. Hence, even considering the current role of RFA as a third-line option, in view of increasing numbers of occurrences detected, the number of potential RFA candidates may see a steep uptrend. In view of all this, it is imperative that interventional radiologists be familiar with the techniques of lung ablation. The aim of this article is to discuss the procedural technique of RFA in the lung and review the current evidence regarding RFA for NSCLC.


Nucleolar proteins and ultrastructure in bovine in vivo developed, in vitro produced, and parthenogenetic cleavage-stage embryos.
In the present study, ribosomal RNA (rRNA) gene activation, monitored through nucleolus development, was studied by autoradiography following (3)H-uridine incubation, transmission electron microscopy, and immunofluorescence confocal laser scanning microscopy of key nucleolar proteins involved in rRNA transcription (topoisomerase I, upstream binding factor, and RNA polymerase I) and processing (fibrillarin, nucleolin, and nucleophosmin) in in vivo developed, in vitro produced, and parthenogenetic bovine embryos. In general, in vivo developed embryos displayed formation of fibrillo-granular nucleoli during the 4th post-fertilization cell cycle. During the previous stages of development, nucleolus precursor bodies (NPBs) were observed. However, on some occasions the initial steps of nucleolus formation were observed already at the 2- and 4-cell stage in cases where such embryos were collected from superovulated animals together with later embryonic stages presenting nucleolar development and autoradiographic labeling. The in vitro produced embryos displayed very synchronous formation of fibrillo-granular nucleoli and autoradiographic labeling during the 4th cell cycle. In vivo developed and in vitro produced embryos displayed allocation of nucleolar proteins to fibrillar and granular compartments of the developing nucleoli during the 4th cell cycle. The parthenogenetic embryos typically displayed formation of fibrillo- granular nucleoli during the 5th cell cycle and autoradiographic labeling was not observed until the morula stage. Moreover, the 1-, 2-, and 4-cell parthenogenetic embryos practically lacked NPBs. On the other hand, parthenogenetic embryos displayed allocation of nucleoar proteins to nuclear entities during the 4th cell cycle. In conclusion, both in vivo developed and in vitro produced bovine embryos displayed activation of transcription and nucleolar development during the 4th cell cycle. However, in vivo developed embryos flushed together with later developmental stages displayed premature activation of these processes. Parthenogenetic bovine embryos, on the other hand, displayed a delayed activation.


C5-methylation of cytosine in B-DNA thermodynamically and kinetically stabilizes BI.
We have examined the backbone dynamics of two alternating purine-pyrimidine dodecamers. One sequence consists of "pure" GC bases; the other one contains 5-methylcytosines. The effect of the methyl groups on the backbone substates BI/BII was investigated by means of molecular dynamics. The methylation influences, on one hand, the transition barrier between BI and BII and, on the other hand, the state of equilibrium. The kinetic consequences are an increase of the DeltaG of Gp5mC steps by 1.5 kcal/mol and a decrease of the DeltaG of 5mCpG steps by 0.8 kcal/mol (compared with the nonmethylated DNA). Thus, the additive group differentiates between the two occurring dinucleotide steps and renders the phosphate of the 5-methylcytosine more rigid, as proposed by experimental studies. The thermodynamic consequences are an increase of the DeltaG of Gp5mC steps by 1.1 kcal/mol and a decrease of the DeltaG of 5mCpG steps by 0.8 kcal/mol. The reason for this shift in equilibrium is still not completely clear on a molecular basis. But we can conclude that the indirect readout of DNA is influenced by methylation.


Measurement of quality of life IV. Use of the SEQOL, QOL5, QOL1, and other global and generic questionnaires.
Quality-of-life (QOL) rating scales can be used to measure and describe the quality of life of a specific population or patient group. Many decisions can be taken and policies implemented when we know more about a group or population. The global quality-of-life concept may help in expressing the objective of the initiatives taken to benefit specific groups. The objective may be that we hope the efforts will increase their quality of life by a certain percentage. This explicit expectation will force the decision makers to stand by their noble intentions. They are obliged to evaluate their efforts and will have to learn something from it. A questionnaire thus constitutes a useful scientific instrument, as databases based on comprehensive and thorough questionnaire surveys that seek to encompass all aspects of life can provide valuable and precise information. The value of such a database depends on the correct use of the questionnaires and this paper examines some examples of how quality-of-life rating scales can be used. We identified at least ten ways to use the quality-of-life questionnaire: describing the quality of life of a population or patient group; formulating an objective for support, treatment, or care; screening or identifying individuals who need treatment; evaluating treatment and care; facilitating communication between physician and patients; involving the patient in the decision-making process; allocating resources; investigating the causal relation between the quality of life and ill health in prospective studies; creating an awareness of the quality of life and health promotion; and helping the practitioner to accumulate knowledge. Enhancing the quality of life is therefore a determining factor in the process of increasing awareness and responsible conduct in relation to the environment, natural resources, the working environment, and the structure of society. Putting the quality of life on the agenda inherently has a constructive and positive effect on the life and functioning of the individual and society.


Dual task during encoding, maintenance, and retrieval in Alzheimer's disease.
Previous dual task studies have demonstrated minimal costs when healthy individuals simultaneously perform two tasks at their own individual ability levels. Conversely, Alzheimer's disease (AD) patients show dual task decrements, but it is unclear whether the problem arises at the encoding, maintenance, and/or retrieval phases of memory. Two experiments combined digit recall and visuo-motor tracking to investigate dual task effects during encoding, maintenance, and/or retrieval for AD patients compared with healthy adults. The demands of each single task were titrated for the ability of each participant. In Experiment 1, the dual task requirement was present throughout both encoding and retrieval of digit recall and the differential dual task effects on a secondary tracking task were examined post-hoc. In Experiment 2, the impact of dual task during encoding only, during maintenance only, and during retrieval only was examined systematically. The findings suggest that the specific AD deficit reflects impairment of a cognitive function that supports the simultaneous performance of two tasks in the healthy brain, particularly during the encoding and retrieval phases of the memory task.


Helicobacter-induced intestinal metaplasia in the stomach correlates with Elk-1 and serum response factor induction of villin.
Chronic Helicobacter pylori infection results in serious sequelae, including atrophy, intestinal metaplasia, and gastric cancer. Intestinal metaplasia in the stomach is defined by the presence of intestine-like cells expressing enterocyte-specific markers, such as villin. In this study, we demonstrate that villin is expressed in intestine-like cells that develop after chronic infection with H. pylori in both human stomach and in a mouse model. Transfection studies were used to identify specific regions of the villin promoter that are inducible by exposure of the cells to H. pylori. We demonstrated that induction of the villin promoter by H. pylori in a human gastric adenocarcinoma cell line (AGS) required activation of the Erk pathway. Elk-1 and the serum response factor (SRF) are downstream transcriptional targets of the Erk pathway. We observed inducible binding of Elk-1 and the SRF after 3 and 24 h of treatment with H. pylori, suggesting that the bacteria alone are sufficient to initiate a cascade of signaling events responsible for villin expression. Thus, H. pylori induction of villin in the stomach correlates with activation and cooperative binding of Elk-1 and the SRF to the proximal promoter of villin.


[Typical changes in the erythron in terminal and post-resuscitation states].
Morphological and biochemical indexes of peripheral erythron compartment have been analysed 1, 3, 7 and 14 days after severe mechanical traumas of various locations in human patients, after 5- and 10-min clinical death in dogs and after rotatory polytrauma in rats. It has been shown that rehabilitation period along with usual anemization is characterized by enhanced penetration of newly formed erythrocytes (hyperchromic, fetal, lipoperoxidation-resistant forms) into the circulation. In pathologies with predominant damage of the central nervous system regenerative response of the bone marrow is less marked and normalization of hematologic parameters is less prompt. As erythron changes always take place in terminal and postresuscitation conditions, irrespective of the etiology and species, they are regarded as a typical pathological process.


Functional analysis of glycoside hydrolase family 8 xylanases shows narrow but distinct substrate specificities and biotechnological potential.
The potential of glycoside hydrolase family (GH) 8 xylanases in biotechnological applications is virtually unexplored. Therefore, the substrate preference and hydrolysis product profiles of two GH8 xylanases were evaluated to investigate their activities and substrate specificities. A GH8 xylanase from an uncultured bacterium (rXyn8) shows endo action but very selectively releases xylotriose from its substrates. It has a higher activity than the Pseudoalteromonas haloplanktis GH8 endo-xylanase (PhXyl) on xylononaose and smaller xylo-oligosaccharides. PhXyl preferably degrades xylan substrates with a high degree of polymerization. It is sterically more hindered by arabinose substituents than rXyn8, producing larger end hydrolysis products. The specificities of rXyn8 and PhXyl differ completely from these of the previously described GH8 xylanases from Bifidobacterium adolescentis (BaRexA) and Bacillus halodurans (BhRex). As reducing-end xylose-releasing exo-oligoxylanases, they selectively release xylose from the reducing end of small xylo-oligosaccharides. The findings of this study show that GH8 xylanases have a narrow substrate specificity, but also one that strongly varies between family members and is distinct from that of GH10 and GH11 xylanases. Structural comparison of rXyn8, PhXyl, BaRexA, and BhRex showed that subtle amino acid changes in the glycon as well as the aglycon subsites probably form the basis of the observed differences between GH8 xylanases. GH8 xylanases, therefore, are an interesting group of enzymes, with potential towards engineering and applications.


Health justice partnerships: initial insights into the delivery of an integrated health and legal service for youth in regional Victoria.
This article presents interim findings from research examining the implementation of a health justice partnership (HJP) focusing on the legal and health needs of regional young people. HJPs provide an innovative service model offering an integrated health and legal service for the community. HJPs are a relatively new service model for Australia, yet the program is well suited to meet the needs of particular population cohorts, including young people and those in regional locations experiencing complex legal issues. Funded by the Victorian Legal Services Board and Commissioner, an HJP in partnership with three organisations was established in a large regional area in Victoria, Australia. Research is being conducted alongside the program to examine its impact on young people, and the implications on practice for staff in the partner organisations. Findings provide preliminary support for the HJP model with a number of young people - from predominantly disadvantaged backgrounds and with varying legal issues - having been referred to the program in the first 6 months. Referrals were received from both partner agencies and external agencies. Initial client and staff survey responses indicate that the legal problem of the young people was affecting how they feel. While these findings provide preliminary support for the HJP further research will offer longer term insights about HJPs within the Australian context, particularly rural and regional settings.


Pharmacogenomics of high-density lipoprotein-cholesterol-raising therapies.
High levels of HDL cholesterol (HDL-C) have traditionally been linked to lower incidence of cardiovascular disease, prompting the search for effective and safe HDL-C raising pharmaceutical agents. Although drugs such as niacin and fibrates represent established therapeutic approaches, HDL-C response to such therapies is variable and heritable, suggesting a role for pharmacogenomic determinants. Multiple genetic polymorphisms, located primarily in genes encoding lipoproteins, cholesteryl ester transfer protein, transporters and CYP450 proteins have been shown to associate with HDL-C drug response in vitro and in epidemiologic studies. However, few of the pharmacogenomic findings have been independently validated, precluding the development of clinical tools that can be used to predict HDL-C response and leaving the goal of personalized medicine to future efforts.


Speech-Language Pathologists' Ratings of Speech Accuracy in Children With Speech Sound Disorders.
Forming accurate and consistent speech judgments can be challenging when working with children with speech sound disorders who produce a large number and varied types of error patterns. Rating scales offer a systematic approach to assessing the whole word rather than individual sounds. Thus, these scales can be an efficient way for speech-language pathologists (SLPs) to monitor treatment progress. This study evaluated the interrater reliability of an existing 3-point rating scale using a large group of SLPs as raters. Utilizing an online platform, 30 SLPs completed a brief training and then rated single words produced by children with typical speech patterns and children with speech sound disorders. Words were closely balanced across the three rating categories of the scale. The interrater reliability of the SLPs ratings to a consensus judgment was examined. The majority of SLPs (87%) reached substantial interrater reliability to a consensus judgment using the 3-point rating scale. Correct productions had the highest interrater reliability. Productions with extensive errors had higher agreement than those with minor errors. Certain error types, such as vowel distortions, were especially challenging for SLPs to judge. This study demonstrated substantial interrater reliability to a consensus judgment among a large majority of 30 SLPs using a 3-point rating. The clinical implications of the findings are discussed along with proposed modifications to the training procedure to guide future research.


ACKR1 favors transcellular over paracellular T-cell diapedesis across the blood-brain barrier in neuroinflammation in vitro.
The migration of CD4+ effector/memory T cells across the blood-brain barrier (BBB) is a critical step in MS or its animal model, EAE. T-cell diapedesis across the BBB can occur paracellular, via the complex BBB tight junctions or transcellular via a pore through the brain endothelial cell body. Making use of primary mouse brain microvascular endothelial cells (pMBMECs) as in vitro model of the BBB, we here directly compared the transcriptome profile of pMBMECs favoring transcellular or paracellular T-cell diapedesis by RNA sequencing (RNA-seq). We identified the atypical chemokine receptor 1 (Ackr1) as one of the main candidate genes upregulated in pMBMECs favoring transcellular T-cell diapedesis. We confirmed upregulation of ACKR1 protein in pMBMECs promoting transcellular T-cell diapedesis and in venular endothelial cells in the CNS during EAE. Lack of endothelial ACKR1 reduced transcellular T-cell diapedesis across pMBMECs under physiological flow in vitro. Combining our previous observation that endothelial ACKR1 contributes to EAE pathogenesis by shuttling chemokines across the BBB, the present data support that ACKR1 mediated chemokine shuttling enhances transcellular T-cell diapedesis across the BBB during autoimmune neuroinflammation.


Increased levels of serum IL-18 are associated with the long-term outcome of severe traumatic brain injury.
A long-lasting neuroinflammatory cascade may lead to the progression of brain damage, favoring neurodegeneration and cognitive impairment in patients with traumatic brain injury (TBI), but the potential mechanisms underlying this sequence of events remain elusive. Here we aimed to evaluate the impact of interleukin (IL)-18, a proinflammatory cytokine elevated in post-acute head injury and associated with neurodegeneration, on the long-term outcome of patients with chronic TBI. The serum content of IL-18 was evaluated in 16 patients with severe TBI, during their rehabilitation phase, and in a matched group of 16 healthy controls. The disability of the enrolled patients was evaluated by means of the Glasgow Outcome Scale, Levels of Cognitive Functioning, and the Disability Rating Scale. The circulating levels of IL-18 were significantly increased in chronic TBI patients, as compared to healthy subjects, and correlated with the patients' cognitive impairment and disability severity. IL-18 may contribute to the long-term outcome and neurodegeneration in TBI patients. Even though further studies are needed to understand the molecular mechanisms underlying the effects of IL-18 on TBI progression and its associated drop in cognitive function, a possible role of this cytokine as a therapeutic target in TBI can be envisaged.


Venous Malformations.
Venous malformations, the most common type of vascular malformation, are slow-flow lesions resulting from disorganized angiogenesis. The International Society for the Study of Vascular Anomalies (ISSVA) classification offers a categorization scheme for venous malformations based on their genetic landscapes and association with congenital overgrowth syndromes. Venous malformations present as congenital lesions and can have broad physiologic and psychosocial sequelae depending on their size, location, growth trajectory, and tissue involvement. Diagnostic evaluation is centered around clinical examination, imaging evaluation with ultrasound and time-resolved magnetic resonance imaging, and genetic testing for more complex malformations. Interventional radiology has emerged as first-line management of venous malformations through endovascular treatment with embolization, while surgery and targeted molecular therapies offer additional therapeutic options. In this review, an updated overview of the genetics and clinical presentation of venous malformations in conjunction with key aspects of diagnostic imaging and treatment are discussed.


Zinc sulfate foot bath for control of ovine foot rot.
A 10% zinc sulfate (ZnSO4) solution was used as a foot bath to control foot rot in pastured lambs. When the foot bath was placed where infected lambs would walk through it each day, clinical evidence of foot rot was greatly reduced within 15 days, and the total number of cases detected after thorough foot trimming was greatly reduced within 30 days. When the foot bath was placed under the salt feeding box, it was not effective. Feeding ZnSO4 at the rate of 0.5 to 0.75 g/head/day was not effective in controlling foot rot. Temperature and moisture conditions were favorable for foot rot transmission throughout the trials.


Minimally invasive aortic valve replacement versus aortic valve replacement through full sternotomy: the Brigham and Women's Hospital experience.
Minimally invasive aortic valve surgery (mini AVR) is a safe and effective treatment option at many hospital centers, but there has not been widespread adoption of the procedure. Critics of mini AVR have called for additional evidence with direct comparison to aortic valve replacement (AVR) via full sternotomy (FS). Our mini AVR approach is through a hemi-sternotomy (HS). We performed a propensity-score matched analysis of all patients undergoing isolated AVR via FS or HS at our institution since 2002, resulting in 552 matched pairs. Baseline characteristics were similar. Operative characteristics, transfusion rates, in-hospital outcomes as well as short and long term survival were compared between groups. Median cardiopulmonary bypass and cross clamp times were shorter in the HS group: 106 minutes [inter-quartile ranges (IQR) 87-135] vs. 124 minutes (IQR 90-169), P≤0.001, and 76 minutes (IQR 63-97) vs. 80 minutes (IQR 62-114), P≤0.005, respectively. HS patients had shorter ventilation times (median 5.7 hours, IQR 3.5-10.3 vs. 6.3 hours, IQR 3.9-11.2, P≤0.022), shorter intensive care unit stay (median 42 hours, IQR 24-71 vs. 45 hours, IQR 24-87, P≤0.039), and shorter hospital length of stay (median 6 days, IQR 5-8 vs. 7 days, IQR 5-10, P≤0.001) compared with the FS group. Intraoperative transfusions were more common in FS group: 27.9% vs. 20.0%, P≤0.003. No differences were seen in short or long term survival, or time to aortic valve re-intervention. Our study confirms the clinical benefits of minimally invasive AVR via HS, which includes decreased transfusion requirements, ventilation times, intensive care unit and hospital length of stay without compromising short and long term survival compared to conventional AVR via FS.


Tolerability of hypertonic and isotonic contrast media injected intravenously. A comparative study in the dog.
We examined the use of isotonic and hypertonic contrast media injected intravenously in the dog from the standpoint of cardiovascular tolerance after right atrial injections performed at 2.56 and 5.12 g I/second. The parameters measured were lead II of the electrocardiograph, heart rate, pulmonary and abdominal arterial pressure, and aortic flow. Three contrast media, ioxitalamate, ioxaglate, and iopamidol (two ionic and one nonionic), were compared, either concentrated (32% iodine) or dilute and isotonic with plasma (ioxaglate 160 mg I/mL and iopamidol 128 mg I/mL). At an injection rate of 5.12 g I/second, iopamidol-128 showed lower electrophysiologic tolerability and caused a higher increase in aortic flow than ioxitalamate 160 or ioxaglate 160. These effects may explain the lower radiographic efficacy observed with iopamidol-128 in previous digital subtraction angiography studies.


CNVannotator: a comprehensive annotation server for copy number variation in the human genome.
Copy number variation (CNV) is one of the most prevalent genetic variations in the genome, leading to an abnormal number of copies of moderate to large genomic regions. High-throughput technologies such as next-generation sequencing often identify thousands of CNVs involved in biological or pathological processes. Despite the growing demand to filter and classify CNVs by factors such as frequency in population, biological features, and function, surprisingly, no online web server for CNV annotations has been made available to the research community. Here, we present CNVannotator, a web server that accepts an input set of human genomic positions in a user-friendly tabular format. CNVannotator can perform genomic overlaps of the input coordinates using various functional features, including a list of the reported 356,817 common CNVs, 181,261 disease CNVs, as well as, 140,342 SNPs from genome-wide association studies. In addition, CNVannotator incorporates 2,211,468 genomic features, including ENCODE regulatory elements, cytoband, segmental duplication, genome fragile site, pseudogene, promoter, enhancer, CpG island, and methylation site. For cancer research community users, CNVannotator can apply various filters to retrieve a subgroup of CNVs pinpointed in hundreds of tumor suppressor genes and oncogenes. In total, 5,277,234 unique genomic coordinates with functional features are available to generate an output in a plain text format that is free to download. In summary, we provide a comprehensive web resource for human CNVs. The annotated results along with the server can be accessed at http://bioinfo.mc.vanderbilt.edu/CNVannotator/.


A bio-inspired approach for the design of a multifunctional robotic end-effector customized for automated maintenance of a reconfigurable vibrating screen.
The development of a robotic-driven maintenance solution capable of automatically maintaining reconfigurable vibrating screen (RVS) machine when utilized in dangerous and hazardous underground mining environment has called for the design of a multifunctional robotic end-effector capable of carrying out all the maintenance tasks on the RVS machine. In view of this, the paper presents a bio-inspired approach which unfolds the design of a novel multifunctional robotic end-effector embedded with mechanical and control mechanisms capable of automatically maintaining the RVS machine. To achieve this, therblig and morphological methodologies (which classifies the motions as well as the actions required by the robotic end-effector in carrying out RVS machine maintenance tasks), obtained from a detailed analogy of how human being (i.e. a machine maintenance manager) will carry out different maintenance tasks on the RVS machine, were used to obtain the maintenance objective functions or goals of the multifunctional robotic end-effector as well as the maintenance activity constraints of the RVS machine that must be adhered to by the multifunctional robotic end-effector during the machine maintenance. The results of the therblig and morphological analyses of five (5) different maintenance tasks capture and classify one hundred and thirty-four (134) repetitive motions and fifty-four (54) functions required in automating the maintenance tasks of the RVS machine. Based on these findings, a worm-gear mechanism embedded with fingers extruded with a hexagonal shaped heads capable of carrying out the "gripping and ungrasping" and "loosening and bolting" functions of the robotic end-effector and an electric cylinder actuator module capable of carrying out "unpinning and hammering" functions of the robotic end-effector were integrated together to produce the customized multifunctional robotic end-effector capable of automatically maintaining the RVS machine. The axial forces ([Formula: see text] and [Formula: see text]), normal forces ([Formula: see text]) and total load [Formula: see text] acting on the teeth of the worm-gear module of the multifunctional robotic end-effector during the gripping of worn-out or new RVS machine subsystems, which are 978.547, 1245.06 and 1016.406 N, respectively, were satisfactory. The nominal bending and torsional stresses acting on the shoulder of the socket module of the multifunctional robotic end-effector during the loosing and tightening of bolts, which are 1450.72 and 179.523 MPa, respectively, were satisfactory. The hammering and unpinning forces utilized by the electric cylinder actuator module of the multifunctional robotic end-effector during the unpinning and hammering of screen panel pins out of and into the screen panels were satisfactory.


A grating-bicoupled plasma-wave photomixer with resonant-cavity enhanced structure.
A novel terahertz plasma-wave photomixer that can improve the conversion gain and terahertz radiation power is proposed and evaluated. The photomixer is based on a high-electron mobility transistor and incorporates doubly interdigitated grating strips for the gate electrodes that periodically localize the 2D plasmons in 100-nm regions with a micron-order interval. A vertical cavity structure is formed in between the top metal grating and a terahertz mirror placed at the backside. The device features electronic tuning of plasmon characteristic frequencies, providing continuously-tunable operation below 1 THz to beyond 10 THz. Frequency-dependent finite-differential time-domain analysis demonstrates that the grating-bicoupled plasmonic structure acts as a broadband terahertz photomixer and antenna and that the vertical cavity structure effectively enhances the conversion gain and radiation power.


Evaluation of a simplified N-acetylcysteine dosing regimen for the treatment of acetaminophen toxicity.
Acetaminophen overdose is the most common pharmaceutical poisoning in the US. The labeled dosing regimen for Acetadote, the only intravenous N-acetylcysteine (IV-NAC) product approved by the Food and Drug Administration (FDA) for treatment of acetaminophen toxicity, is a complex 3-step process that produces frequent medication errors. We have been using an off-label, uncomplicated dosing regimen consisting of a standard preparation of IV-NAC 30 g in 1 L of 5% dextrose in water, with a 150-mg/kg loading dose administered over 1 hour followed by an infusion of 14 mg/kg/h for 20 hours. To evaluate the frequency of medication errors, resolution of hepatotoxicity, and tolerability of the protocol used in our institution for treatment of acetaminophen toxicity. This single-center, retrospective chart review evaluated patients receiving IV-NAC for acetaminophen toxicity from August 2006 to August 2008. Charts were reviewed for prescribing practices, dosing errors, and clinical outcomes. Among 70 patients who met inclusion criteria, 35 medication errors occurred, including 22 administration errors and 13 protocol initiation errors. The frequency of administration errors was 13.5 errors per 100 administration interventions. Loading dose errors were most common with 11 rate-related and 8 dose-related errors. Interruptions longer than 60 minutes occurred in only 3 patients. No adverse outcomes were associated with medication errors. The mean (SD) duration of therapy was 25.6 hours (n = 60 pts. [17.8], range 1-76.5), and mean length of stay was 2.99 days ([3.82], range 0.1-25.7). All patients with hepatotoxicity (aspartate aminotransferase >1000 units/L) due to acute acetaminophen toxicity had resolution of the toxicity and were successfully discharged. This single intravenous bag protocol is effective and well tolerated, and there is infrequent interruption of therapy. The overall rate of administration errors is similar to that in reports on the FDA regimen; thus, our protocol may be an acceptable alternative.


Prothrombin complex proteins as cofactors in platelet aggregation. I. Inhibition of aggregation by antiserum.
Data are presented to support the concept that the vitamin K-dependent coagulation factors are adsorbed onto the platelet membrane and that these make up part of the "plasma atmosphere" necessary for the aggregation of platelets by various agents. Together with evidence that other coagulation factors also are part of the plasma atmosphere, it is suggested that the aggregation reaction is part of the coagulation sequence. The immunologic approach to demonstrating constituents of the platelet membrane promises to be a highly specific technique for studying further the constituents of the platelet membrane and their reactions in hemostasis.


Visualization of antitumor treatment by means of fluorescence molecular tomography with an annexin V-Cy5.5 conjugate.
In vivo imaging of treatment responses at the molecular level could have a significant impact on the speed of drug discovery and ultimately lead to personalized medicine. Strong interest has been shown in developing quantitative fluorescence-based technologies with good molecular specificity and sensitivity for noninvasive 3D imaging through tissues and whole animals. We show herein that tumor response to chemotherapy can be accurately resolved by fluorescence molecular tomography (FMT) with a phosphatidylserine-sensing fluorescent probe based on modified annexins. We observed at least a 10-fold increase of fluorochrome concentration in cyclophosphamide-sensitive tumors and a 7-fold increase of resistant tumors compared with control studies. FMT is an optical imaging technique developed to overcome limitations of commonly used planar illumination methods and demonstrates higher quantification accuracy validated by histology. It is further shown that a 3-fold variation in background absorption heterogeneity may yield 100% errors in planar imaging but only 20% error in FMT, thus confirming tomographic imaging as a preferred tool for quantitative investigations of fluorescent probes in tissues. Tomographic approaches are found essential for small-animal optical imaging and are potentially well suited for clinical drug development and monitoring.


Influence of social deprivation on provision of bariatric surgery: 10-year comparative ecological study between two UK specialist centres.
To investigate the effect of residential location and socioeconomic deprivation on the provision of bariatric surgery. Retrospective cross-sectional ecological study. Patients resident local to one of two specialist bariatric units, in different regions of the UK, who received obesity surgery between 2003 and 2013. Demographic data were collected from prospectively collected databases. Index of Multiple Deprivation (IMD 2010) was used as a measure of socioeconomic status. Obesity prevalences were obtained from Public Health England (2006). Patients were split into three IMD tertiles (high, median, low) and also tertiles of time. A generalised linear model was generated for each time period to investigate the effect of socioeconomic deprivation on the relationship between bariatric case count and prevalence of obesity. We used these to estimate surgical intervention provided in each population in each period at differing levels of deprivation. Data were included from 1163 bariatric cases (centre 1-414, centre 2-749). Incidence rate ratios (IRRs) were calculated to measure the associations between predictor and response variables. Associations were highly non-linear and changed over the 10-year study period. In general, the relationship between surgical case volume and obesity prevalence has weakened over time, with high volumes becoming less associated with prevalence of obesity. As bariatric services have matured, the associations between demand and supply factors have changed. Socioeconomic deprivation is not apparently a barrier to service provision more recently, but the positive relationships between obesity and surgical volume we would expect to find are absent. This suggests that interventions are not being taken up in the areas of need. We recommend a more detailed national analysis of the relationship between supply side and demand side factors in the provision of bariatric surgery.


Management and outcome of peptic ulcers or erosions in patients receiving a combination of aspirin plus clopidogrel.
This multicenter retrospective study investigated the management and outcome of patients with peptic ulcer/erosion-related aspirin and clopidogrel (A + C) cotherapy. From January 2002 to September 2006, patients with endoscopically proven peptic ulcers/erosions after receiving A + C cotherapy were analyzed. This group consisted of 106 patients (age, 69.3 +/- 11.7 years). Ulcers/erosions developed in 27 patients during hospitalization for cardiac events and in 79 patients after hospital discharge. Of 27 patients hospitalized for acute cardiac events, gastrointestinal (GI) bleeding and dyspepsia occurred in 24 and three, respectively. The most common lesion was gastric ulcer. Of 79 discharged patients, GI bleeding and dyspepsia occurred in 64 and 15, respectively. The most common bleeding and dyspeptic lesions were gastric ulcer and gastritis, respectively. Overall, 17 patients underwent endoscopic hemostasis all successfully. A + C cotherapy was continued in 57 patients for a median (interquartile range) of 3.0 (6.2) months. Most were coprescribed a proton pump inhibitor (PPI) (53, 93%). No recurrent GI bleeding was observed. After A + C cotherapy, gastric ulcer or gastritis were the most common endoscopic lesions. The combination of a PPI and endoscopic treatment for ulcer bleeding was highly successful. After patient stabilization, continuation of A + C cotherapy with a PPI appears to be safe.


Temporary threshold shift induced by music.
Three groups of young people, in all 10 males and 11 females, with different music listening habits listened to their own choice of music from a portable cassette player for one hour. The sound pressure level from the music was measured with a probe-provided miniature microphone in the external auditory canal close to the tympanic membrane. The temporary threshold shift induced by the music, as well as by 1/3-octave band noise, was registered with Békéky audiometry. The females had significantly more temporary threshold shift than the males after noise-exposure. Most subjects had only discrete temporary threshold shifts after one hour of listening to music, in spite of 91-97 dB listening levels. There were no significant differences in listening levels or music-induced threshold shifts between genders, although such differences were found between groups with different listening habits.


"Covalent affinity" purification of ubiquitin-activating enzyme.
We have previously described an enzyme that activates ubiquitin, the heat-stable polypeptide of the ATP-dependent proteolytic system from reticulocytes (Ciechanover, A., Heller, H., Katz-Etzion, R., and Hershko, A. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 761-765). It carries out ubiquitin-dependent PPi-ATP and AMP-ATP exchange reactions and binds to the activated polypeptide by a thiolester linkage. We describe here a procedure for the purification of this enzyme by its binding to ubiquitin-Sepharose. Binding of the enzyme to the affinity column requires ATP and Mg2+, and bound enzyme cannot be displaced by high salt but can be eluted by raising the pH, by increased concentrations of a thiol compound, or by the joint supplementation of AMP and pyrophosphate. Another form of the enzyme that cannot carry out AMP-ATP exchange (but catalyzes ubiquitin-dependent PPi-ATP exchange) does not bind to the affinity column. These data indicate that a covalent, possibly thiolester intermediate, is formed between the activating enzyme and Sepharose-bound ubiquitin. It is suggested designating this procedure of enzyme isolation "covalent affinity" chromatography. The purified enzyme has an apparent Mr = 210,000 and appears to be composed of two subunits of Mr = 105, 000. ATP-dependent binding of ubiquitin to the purified enzyme and to its subunit is demonstrated.


Interleukin-10 improves stroke outcome by controlling the detrimental Interleukin-17A response.
Lymphocytes have dichotomous functions in ischemic stroke. Regulatory T cells are protective, while IL-17A from innate lymphocytes promotes the infarct growth. With recent advances of T cell-subtype specific transgenic mouse models it now has become possible to study the complex interplay of T cell subpopulations in ischemic stroke. In a murine model of experimental stroke we analyzed the effects of IL-10 on the functional outcome for up to 14 days post-ischemia and defined the source of IL-10 in ischemic brains based on immunohistochemistry, flow cytometry, and bone-marrow chimeric mice. We used neutralizing IL-17A antibodies, intrathecal IL-10 injections, and transgenic mouse models which harbor a deletion of the IL-10R on distinct T cell subpopulations to further explore the interplay between IL-10 and IL-17A pathways in the ischemic brain. We demonstrate that IL-10 deficient mice exhibit significantly increased infarct sizes on days 3 and 7 and enlarged brain atrophy and impaired neurological outcome on day 14 following tMCAO. In ischemic brains IL-10 producing immune cells included regulatory T cells, macrophages, and microglia. Neutralization of IL-17A following stroke reversed the worse outcome in IL-10 deficient mice and intracerebral treatment with recombinant IL-10 revealed that IL-10 controlled IL-17A positive lymphocytes in ischemic brains. Importantly, IL-10 acted differentially on αβ and γδ T cells. IL-17A producing CD4+ αβ T cells were directly controlled via their IL-10-receptor (IL-10R), whereas IL-10 by itself had no direct effect on the IL-17A production in γδ T cells. The control of the IL-17A production in γδ T cells depended on an intact IL10R signaling in regulatory T cells (Tregs). Taken together, our data indicate a key function of IL-10 in restricting the detrimental IL-17A-signaling in stroke and further supports that IL-17A is a therapeutic opportunity for stroke treatment.


ICTV Virus Taxonomy Profile: Pneumoviridae.
The family Pneumoviridae comprises large enveloped negative-sense RNA viruses. This taxon was formerly a subfamily within the Paramyxoviridae, but was reclassified in 2016 as a family with two genera, Orthopneumovirus and Metapneumovirus. Pneumoviruses infect a range of mammalian species, while some members of the Metapneumovirus genus may also infect birds. Some viruses are specific and pathogenic for humans, such as human respiratory syncytial virus and human metapneumovirus. There are no known vectors for pneumoviruses and transmission is thought to be primarily by aerosol droplets and contact. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the taxonomy of the Pneumoviridae, which is available at www.ictv.global/report/pneumoviridae.


Localisation and interaction of the protein components of the yeast 2 mu circle plasmid partitioning system suggest a mechanism for plasmid inheritance.
Replicating plasmids are highly unstable in yeast, because they are retained in mother cells. The 2 mu circle plasmid overcomes this maternal inheritance bias by using a partitioning system that involves the plasmid encoded proteins Rep1p and Rep2p, and the cis-acting locus STB. It is thus widely exploited as a cloning vehicle in yeast. However, little is known about the cellular or molecular mechanisms by which effective partitioning is achieved, and models of both free diffusion and plasmid localisation have been proposed. Here we show that Rep1p and Rep2p proteins interact to form homo- and hetero-complexes in vitro. In vivo, Rep1p and Rep2p are shown to be nuclear proteins, exhibiting sub-nuclear concentration in distinct foci. The number of foci appears constant regardless of plasmid copy number and cell ploidy level. Before cell division, the number of foci increases, and we observe approximately equal allocation of foci to mother and daughter cell nuclei. We show that whereas Rep2p expressed alone is found exclusively in the nucleus, Rep1p requires the presence of Rep2p for effective nuclear localisation. High levels of 2 mu plasmid induce a multiple-budded elongated cell phenotype, which we show can be phenocopied by overexpression of both REP1 and REP2 together but not alone. Taken together, these results suggest that Rep1p and Rep2p interact in vivo, and occupy defined nuclear sites that are allocated to both mother and daughter nuclei during division. We propose a model for 2 mum plasmid partitioning based on these results, involving the association of plasmid DNA with specific, segregated subnuclear sites.


Discontinuation of anticytomegalovirus therapy in patients with HIV infection and cytomegalovirus retinitis.
Persons with cytomegalovirus (CMV) retinitis and acquired immunodeficiency syndrome (AIDS) have required lifelong anti-CMV therapy to prevent the progression of retinal disease and subsequent loss of vision. To determine whether patients who were taking highly active antiretroviral therapy (HAART) and who had stable CMV retinitis could safely discontinue anti-CMV therapy without reactivation of their retinitis or increase in human immunodeficiency virus (HIV) viral load. Prospective nonrandomized interventional trial performed from July 1997 to August 1999. Clinical Center of the National Institutes of Health, Bethesda, Md. Fourteen patients with stable CMV retinitis and HIV infection and CD4+ cell counts higher than 0.1 5 x 10(9)/L and being treated with systemic anti-CMV medications and HAART. Discontinuation of specific anti-CMV therapy. Reactivation of CMV retinitis, development of extraocular CMV infection, detection of CMV in blood and urine, HIV burden, immunologic function, quality of life, morbidity, and mortality. Twelve (89.7%) of 14 patients had evidence of immune recovery uveitis before anti-CMV drugs were discontinued. No patient had reactivation of CMV retinitis or development of extraocular CMV disease during mean follow-up of 16.4 months (range, 8.3-22.0 months) without anti-CMV therapy. Human immunodeficiency viral load remained stable following cessation of anti-CMV medications. Blood and urine assays for CMV were briefly positive in 9 patients but did not predict reactivation of CMV disease. Worsening immune recovery uveitis was associated with a substantial (>3 lines) vision loss in 3 patients. Maintenance anti-CMV medications were safely stopped in those patients who had stable CMV retinitis and elevated CD4+ cell counts and who were taking HAART. The study demonstrates that immune recovery following potent antiretroviral therapy is effective in controlling a major opportunistic infection, even in patients with a history of severe immunosuppression.


H2S induces a suspended animation-like state in mice.
Mammals normally maintain their core body temperature (CBT) despite changes in environmental temperature. Exceptions to this norm include suspended animation-like states such as hibernation, torpor, and estivation. These states are all characterized by marked decreases in metabolic rate, followed by a loss of homeothermic control in which the animal's CBT approaches that of the environment. We report that hydrogen sulfide can induce a suspended animation-like state in a nonhibernating species, the house mouse (Mus musculus). This state is readily reversible and does not appear to harm the animal. This suggests the possibility of inducing suspended animation-like states for medical applications.