The Development of Practice Recommendations for Drug-Disease Interactions by Literature Review and Expert Opinion.
Drug-disease interactions negatively affect the benefit/risk ratio of drugs for specific populations. In these conditions drugs should be avoided, adjusted, or accompanied by extra monitoring. The motivation for many drug-disease interactions in the Summary of Product Characteristics (SmPC) is sometimes insufficiently supported by (accessible) evidence. As a consequence the translation of SmPC to clinical practice may lead to non-specific recommendations. For the translation of this information to the real world, it is necessary to evaluate the available knowledge about drug-disease interactions, and to formulate specific recommendations for prescribers and pharmacists. The aim of this paper is to describe a standardized method how to develop practice recommendations for drug-disease interactions by literature review and expert opinion. The development of recommendations for drug-disease interactions will follow a six-step plan involving a multidisciplinary expert panel (1). The scope of the drug-disease interaction will be specified by defining the disease and by describing relevant effects of this drug-disease interaction. Drugs possibly involved in this drug-disease interaction are selected by checking the official product information, literature, and expert opinion (2). Evidence will be collected from the official product information, guidelines, handbooks, and primary literature (3). Study characteristics and outcomes will be evaluated and presented in standardized reports, including preliminary conclusions on the clinical relevance and practice recommendations (4). The multidisciplinary expert panel will discuss the reports and will either adopt or adjust the conclusions (5). Practice recommendations will be integrated in clinical decision support systems and published (6). The results of the evaluated drug-disease interactions will remain up-to-date by screening new risk information, periodic literature review, and (re)assessments initiated by health care providers. The practice recommendations will result in advices for specific DDSI. The content and considerations of these DDSIs will be published and implemented in all Clinical Decision Support Systems in the Netherlands. The recommendations result in professional guidance in the context of individual patient care. The professional will be supported in the decision making in concerning pharmacotherapy for the treatment of a medical problem, and the clinical risks of the proposed medication in combination with specific diseases.


Severe energy deficit upregulates leptin receptors, leptin signaling, and PTP1B in human skeletal muscle.
In obesity, leptin receptors (OBR) and leptin signaling in skeletal muscle are downregulated. To determine whether OBR and leptin signaling are upregulated with a severe energy deficit, 15 overweight men were assessed before the intervention (PRE), after 4 days of caloric restriction (3.2 kcal·kg body wt-1·day-1) in combination with prolonged exercise (CRE; 8 h walking + 45 min single-arm cranking/day) to induce an energy deficit of ~5,500 kcal/day, and following 3 days of control diet (isoenergetic) and reduced exercise (CD). During CRE, the diet consisted solely of whey protein (n = 8) or sucrose (n = 7; 0.8 g·kg body wt-1·day-1). Muscle biopsies were obtained from the exercised and the nonexercised deltoid muscles and from the vastus lateralis. From PRE to CRE, serum glucose, insulin, and leptin were reduced. OBR expression was augmented in all examined muscles associated with increased maximal fat oxidation. Compared with PRE, after CD, phospho-Tyr1141OBR, phospho-Tyr985OBR, JAK2, and phospho-Tyr1007/1008JAK2 protein expression were increased in all muscles, whereas STAT3 and phospho-Tyr705STAT3 were increased only in the arms. The expression of protein tyrosine phosphatase 1B (PTP1B) in skeletal muscle was increased by 18 and 45% after CRE and CD, respectively (P < 0.05). Suppressor of cytokine signaling 3 (SOCS3) tended to increase in the legs and decrease in the arm muscles (ANOVA interaction: P < 0.05). Myosin heavy chain I isoform was associated with OBR protein expression (r = -0.75), phospho-Tyr985OBR (r = 0.88), and phospho-Tyr705STAT3/STAT3 (r = 0.74). In summary, despite increased PTP1B expression, skeletal muscle OBR and signaling are upregulated by a severe energy deficit with greater response in the arm than in the legs likely due to SOCS3 upregulation in the leg muscles.NEW & NOTEWORTHY This study shows that the skeletal muscle leptin receptors and their corresponding signaling cascade are upregulated in response to a severe energy deficit, contributing to increase maximal fat oxidation. The responses are more prominent in the arm muscles than in the legs but partly blunted by whey protein ingestion and high volume of exercise. This occurs despite an increase of protein tyrosine phosphatase 1B protein expression, a known inhibitor of insulin and leptin signaling.


Increased risk of endocrine autoimmunity in first-degree relatives of patients with autoimmune Addison's disease.
Autoimmune conditions tend to cluster in subjects with Addison's disease (AD) and probably also among their relatives. The aim of the study was to estimate the frequency of the endocrine gland-specific autoantibodies in first-degree relatives of patients with AD. Autoantibodies were investigated in 113 family members using RIA and ELISA assays. The control group comprised 143 age-matched volunteers. Autoimmune diseases were diagnosed in 38.1% relatives. Hashimoto's thyroiditis was found in 20.3%, Graves' disease in 8.0%, vitiligo and type 1 diabetes in 3.5%, whereas AD, rheumatoid arthritis and atrophic gastritis with pernicious anaemia in 2.7% each. All studied antibodies except for islet antigen-2 (P = 0.085) were significantly more frequent in AD relatives than in controls (P < 0.05). Antibodies to 21-hydroxylase were detected in 6.2% relatives, thyroid peroxidase in 28.3%, thyroglobulin in 19.5%, glutamic acid decarboxylase in 8.0%, and zinc transporter-8 in 7.1%. Two and more autoantibodies were detected in 18.6% subjects. Significant gender difference was revealed only for aTPO, more common in female relatives (P = 0.014; OR: 3.16; 95% CI: 1.23-8.12). Circulating autoantibodies were found more frequently in the relatives of affected males (P = 0.008; OR: 3.31; 95% CI: 1.33-8.23), and in family members of patients with polyendocrine autoimmunity (P = 0.009; OR: 3.55; 95% CI: 1.31-9.57). This study provides evidence of increased susceptibility for the endocrine autoimmunity, especially thyroid disease, in close relatives of patients with AD. Relatives of the male AD patients and of those with autoimmune polyendocrine syndrome are at particular risk and should undergo periodic screening for autoimmune endocrine disorders.


Nosocomial infection indicators in Australian hospitals: assessment according to hospital characteristics.
The relationship of bed size and hospital type (private or public) was studied using Hospital-Wide Medical Indicator data on nosocomial infections submitted to the Australian Council on Healthcare Standards Care Evaluation Program by hospitals presenting voluntarily for accreditation in 1993. The aim was to determine if this process could simplify the establishment of hospital peer groups for comparison of risk in the absence of knowledge of patient illness severity indices. After adjusting for potential confounders in a logistic model, hospital type was found to be a significant predictor for the occurrence of infection in clean and contaminated wounds. Bed size was a significant predictor for the occurrence of hospital-acquired bacteraemia in private and public hospitals. The increase in the risk of developing hospital acquired bacteraemia with increasing number of beds was significant as a trend (P < 0.0001) in private as well as public hospitals. The results suggest that hospital type and bed size are initial indices for 'flagging' peer group variation and prompting a more detailed internal review.


Maximum activity of recombinant ribulose 1,5-bisphosphate carboxylase/oxygenase of Anabaena sp. strain CA requires the product of the rbcX gene.
Filamentous cyanobacteria of the genus Anabaena contain a unique open reading frame, rbcX, which is juxtaposed and cotranscribed with the genes (rbcL and rbcS) encoding form I ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO). Plasmid constructions containing the genes from Anabaena sp. strain CA were prepared, and expression studies in Escherichia coli indicated that the product of the rbcX gene mimicked the ability of chaperonin proteins to facilitate the proper folding of recombinant RubisCO proteins. The purified recombinant Anabaena sp. strain CA RubisCO, much like the RubisCO enzymes from other cyanobacteria, was shown not to undergo inhibition of activity during a time course experiment, and the properties of this chaperoned recombinant protein appear to be consistent with those of the enzyme isolated from the native organism.


Understanding the Twisted Structure of Amyloid Fibrils via Molecular Simulations.
Accumulation and aggregation of amyloid are associated with the pathogenesis of many human diseases, such as Alzheimer's disease and Type 2 diabetes mellitus. Therefore, a quantitative understanding of the molecular mechanisms causing different aggregated structures and biomechanical properties of amyloid fibrils could shed some light into the progression of these diseases. In this work, we develop coarse-grained molecular dynamics (CGMD) models to simulate the dynamic self-assembly of two types of amyloids (amylin and amyloid β (Aβ)). We investigate the structural and mechanical properties of different types of aggregated amyloid fibrils. Our simulations demonstrate that amyloid fibrils could result from longitudinal growth of protofilament bundles, confirming one of the hypotheses on the fibril formation. In addition, we find that the persistence length of amylin fibrils increases concurrently with their pitch length, suggesting that the bending stiffness of amylin fibrils becomes larger when the amylin fibrils are less twisted. Similar results are observed for Aβ fibrils. These findings quantify the connection between the structural and the biomechanical properties of the fibrils. The CGMD models developed in this work can be potentially used to examine efficacy of anti-aggregation drugs, which could help in developing new treatments.


Cohort Profile: The Mexican Health and Aging Study (MHAS).
The Mexican Health and Aging Study (MHAS) was designed to prospectively evaluate the impact of disease on the health, function and mortality of adults over the age of 50 in both urban and rural areas of Mexico. The overall goal of the study is to examine the ageing process and its disease and disability burden in a large representative panel of older Mexicans, using a wide socioeconomic perspective. The study protocols and survey instruments are highly comparable to the U.S. Health and Retirement Study (HRS).The MHAS 2001 baseline is a nationally and urban-rural representative survey of individuals born in 1951 or earlier. Three waves of data have been collected so far: baseline in 2001 and follow-ups in 2003 and 2012. In 2012, the study added a representative sample of the population from the 1952-62 birth cohorts. A fourth wave will be collected in 2015.The data files and documentation are available free of charge at the study website [www.MHASweb.org] in English and [www.ENASEM.org] in Spanish.


A physiotherapist-delivered integrated exercise and pain coping skills training intervention for individuals with knee osteoarthritis: a randomised controlled trial protocol.
Knee osteoarthritis (OA) is a prevalent chronic musculoskeletal condition with no cure. Pain is the primary symptom and results from a complex interaction between structural changes, physical impairments and psychological factors. Much evidence supports the use of strengthening exercises to improve pain and physical function in this patient population. There is also a growing body of research examining the effects of psychologist-delivered pain coping skills training (PCST) particularly in other chronic pain conditions. Though typically provided separately, there are symptom, resource and personnel advantages of exercise and PCST being delivered together by a single healthcare professional. Physiotherapists are a logical choice to be trained to deliver a PCST intervention as they already have expertise in administering exercise for knee OA and are cognisant of the need for a biopsychosocial approach to management. No studies to date have examined the effects of an integrated exercise and PCST program delivered solely by physiotherapists in this population. The primary aim of this multisite randomised controlled trial is to investigate whether an integrated 12-week PCST and exercise treatment program delivered by physiotherapists is more efficacious than either program alone in treating pain and physical function in individuals with knee OA. This will be an assessor-blinded, 3-arm randomised controlled trial of a 12-week intervention involving 10 physiotherapy visits together with home practice. Participants with symptomatic and radiographic knee OA will be recruited from the community in two cities in Australia and randomized into one of three groups: exercise alone, PCST alone, or integrated PCST and exercise. Randomisation will be stratified by city (Melbourne or Brisbane) and gender. Primary outcomes are overall average pain in the past week measured by a Visual Analogue Scale and physical function measured by the Western Ontario and McMaster Universities Osteoarthritis Index subscale. Secondary outcomes include global rating of change, muscle strength, functional performance, physical activity levels, health related quality of life and psychological factors. Measurements will be taken at baseline and immediately following the intervention (12 weeks) as well as at 32 weeks and 52 weeks to examine maintenance of any intervention effects. Specific assessment of adherence to the treatment program will also be made at weeks 22 and 42. Relative cost-effectiveness will be determined from health service usage and outcome data. The findings from this randomised controlled trial will provide evidence for the efficacy of an integrated PCST and exercise program delivered by physiotherapists in the management of painful and functionally limiting knee OA compared to either program alone. Australian New Zealand Clinical Trials Registry reference number: ACTRN12610000533099.


[Effect of prolonged chronic stimulation of positive emotionogenic zones of the hypothalamus on blood lipid levels and arterial pressure].
Chronic experiments on rabbits with electrodes implanted into positive affective zones of the hypothalamus, which provided the reaction of self-stimulation during experimentation have shown that prolonged, discrete (over 2 hours daily), chronic (over 2 months stimulation of these zones in unrestrained animals does not cause stable changes in the content of blood lipids but lead to a significant lowering of arterial blood pressure. Continued stimulation (for one month more) together with daily feeding of rabbits with small doses of cholesterol (60 mg/kg bw) produced the same effect as the stimulation of the negative emotiogenic zones, namely, a greater increase of alimentary hypercholesterolemia than in animals of two control groups (non-operated and with implanted electrodes), as well as arterial blood pressure elevation.


Use of Multiple Linear Regression Models for Setting Water Quality Criteria for Copper: A Complementary Approach to the Biotic Ligand Model.
Biotic Ligand Models (BLMs) for metals are widely applied in ecological risk assessments and in the development of regulatory water quality guidelines in Europe, and in 2007 the United States Environmental Protection Agency (USEPA) recommended BLM-based water quality criteria (WQC) for Cu in freshwater. However, to-date, few states have adopted BLM-based Cu criteria into their water quality standards on a state-wide basis, which appears to be due to the perception that the BLM is too complicated or requires too many input variables. Using the mechanistic BLM framework to first identify key water chemistry parameters that influence Cu bioavailability, namely dissolved organic carbon (DOC), pH, and hardness, we developed Cu criteria using the same basic methodology used by the USEPA to derive hardness-based criteria but with the addition of DOC and pH. As an initial proof of concept, we developed stepwise multiple linear regression (MLR) models for species that have been tested over wide ranges of DOC, pH, and hardness conditions. These models predicted acute Cu toxicity values that were within a factor of ±2 in 77% to 97% of tests (5 species had adequate data) and chronic Cu toxicity values that were within a factor of ±2 in 92% of tests (1 species had adequate data). This level of accuracy is comparable to the BLM. Following USEPA guidelines for WQC development, the species data were then combined to develop a linear model with pooled slopes for each independent parameter (i.e., DOC, pH, and hardness) and species-specific intercepts using Analysis of Covariance. The pooled MLR and BLM models predicted species-specific toxicity with similar precision; adjusted R2 and R2 values ranged from 0.56 to 0.86 and 0.66-0.85, respectively. Graphical exploration of relationships between predicted and observed toxicity, residuals and observed toxicity, and residuals and concentrations of key input parameters revealed many similarities and a few key distinctions between the performances of the two models. The pooled MLR model was then applied to the species sensitivity distribution to derive acute and chronic criteria equations similar in form to the USEPA's current hardness-based criteria equations but with DOC, pH, and hardness as the independent variables. Overall, the MLR is less responsive to DOC than the BLM across a range of hardness and pH conditions but more responsive to hardness than the BLM. Additionally, at low and intermediate hardness, the MLR model is less responsive than the BLM to pH, but the two models respond comparably at high hardness. The net effect of these different response profiles is that under many typical water quality conditions, MLR- and BLM-based criteria are quite comparable. Indeed, conditions where the two models differ most (high pH/low hardness and low pH/high hardness) are relatively rare in natural aquatic systems. We suggest that this MLR-based approach, which includes the mechanistic foundation of the BLM but is also consistent with widely accepted hardness-dependent WQC in terms of development and form, may facilitate adoption of updated state-wide Cu criteria that more accurately account for the parameters influencing Cu bioavailability than current hardness-based criteria.


A New Drug Discovery Platform: Application to DNA Polymerase Eta and Apurinic/Apyrimidinic Endonuclease 1.
The ability to quickly discover reliable hits from screening and rapidly convert them into lead compounds, which can be verified in functional assays, is central to drug discovery. The expedited validation of novel targets and the identification of modulators to advance to preclinical studies can significantly increase drug development success. Our SaXPyTM ("SAR by X-ray Poses Quickly") platform, which is applicable to any X-ray crystallography-enabled drug target, couples the established methods of protein X-ray crystallography and fragment-based drug discovery (FBDD) with advanced computational and medicinal chemistry to deliver small molecule modulators or targeted protein degradation ligands in a short timeframe. Our approach, especially for elusive or "undruggable" targets, allows for (i) hit generation; (ii) the mapping of protein-ligand interactions; (iii) the assessment of target ligandability; (iv) the discovery of novel and potential allosteric binding sites; and (v) hit-to-lead execution. These advances inform chemical tractability and downstream biology and generate novel intellectual property. We describe here the application of SaXPy in the discovery and development of DNA damage response inhibitors against DNA polymerase eta (Pol η or POLH) and apurinic/apyrimidinic endonuclease 1 (APE1 or APEX1). Notably, our SaXPy platform allowed us to solve the first crystal structures of these proteins bound to small molecules and to discover novel binding sites for each target.


Complement factor 5 (C5) p.A252T mutation is prevalent in, but not restricted to, sub-Saharan Africa: implications for the susceptibility to meningococcal disease.
Complement C5 deficiency (C5D) is a rare primary immunodeficiency associated with recurrent infections, particularly meningitis, by Neisseria species. To date, studies to elucidate the molecular basis of hereditary C5D have included fewer than 40 families, and most C5 mutations (13 of 17) have been found in single families. However, the recently described C5 p.A252T mutation is reported to be associated with approximately 7% of meningococcal disease cases in South Africa. This finding raises the question of whether the mutation may be prevalent in other parts of Africa or other continental regions. The aim of this study was to investigate the prevalence of C5 p.A252T in Africa and other regions and discuss the implications for prophylaxis against meningococcal disease. In total, 2710 samples from healthy donors within various populations worldwide were analysed by quantitative polymerase chain reaction (qPCR) assay to detect the C5 p.A252T mutation. Eleven samples were found to be heterozygous for p.A252T, and nine of these samples were from sub-Saharan African populations (allele frequency 0·94%). Interestingly, two other heterozygous samples were from individuals in populations outside Africa (Israel and Pakistan). These findings, together with data from genomic variation databases, indicate a 0·5-2% prevalence of the C5 p.A252T mutation in heterozygosity in sub-Saharan Africa. Therefore, this mutation may have a relevant role in meningococcal disease susceptibility in this geographical area.


Application of the Robotic-Assisted Digital Exoscope for Resection of Posterior Fossa Tumors in Adults: A Series of 45 Cases.
Complete safe resection is the goal when pursuing surgical treatment for posterior fossa (PF) tumors. Efforts have led to the development of the exoscope that delineates tumors from non-neoplastic brain. This investigation aims to assess patient outcomes where PF tumor resection is performed with the exoscope by a retromastoid or suboccipital approach. A retrospective analysis was conducted for patients with PF tumors who underwent exoscope resection from 2017 to 2022. Patient demographics, clinical, operative, and outcome findings were collected. Extent of resection studies were also performed. Associations between perioperative data, discharge disposition, progression-free survival (PFS), and overall survival (OS) were evaluated. A total of 45 patients (22 male patients) with a median age of 57 years were assessed. Eighteen (40%) and 27 patients (60%) were diagnosed with malignant and benign tumors, respectively. Tumor neurovascular involvement was found in 28 patients (62%). Twenty-four (53%) and 20 (44%) tumors formed in the cerebellum and cerebellopontine angle cistern, respectively. One tumor (2%) was found in the cervicomedullary junction. The mean extent of resection was 96.7% for benign and malignant tumors. The PFS and OS rate at 6 months (PFS6, OS6) was 89.7% and 95.5%, respectively. Neurological complications included sensory loss and motor deficit, with 11 patients reporting no postoperative symptoms. Of the neurological complications, 14 were temporary and 9 were permanent. The exoscope is an effective intraoperative visualization tool for delineating PF tumors. In our series, we achieved low postoperative tumor volumes and a high gross total resection rate.


The Mission of Safety Net Hospitals: Charity or Equity?
The traditional mission of safety net hospitals has been charity, providing the best healthcare for all individuals no matter their ability to pay. The focus has been on vulnerable populations that are low-income, uninsured, and other upstream circumstances that manifest downstream as poor health, poor health outcomes, and repeated high-cost interventions that fail to break cycles of perpetual health instability. Safety net hospitals are committed to serving their populations, even if only temporarily, through provision of subsidies and filling gaps that exist in patients' lives. These interventions do not lead to the elimination of gaps, hence cyclical health instability persists. It is a new day in healthcare and what it means for people to be well. The focus is on improving health outcomes by addressing root causes of health instability such as unstable housing, income, education, and access to affordable healthy foods. This gives us pause to reflect on the traditional mission of safety net hospitals and the impact of charity in isolation. Are safety net hospitals missing an opportunity to mitigate and eliminate perpetual health instability? Can they shift the paradigm of healthcare for vulnerable populations to alter their quality-of-life course? To move forward, safety net hospitals have to change their mind set and existing narratives about what is possible for vulnerable populations to achieve. These historic giants in healthcare have an opportunity to use their assets and employ a methodology of disruption and innovation to shift the mission of safety net healthcare from charity to equity.


Pubic inguinal pain syndrome: the so-called sports hernia.
The "sportsman's hernia" commonly presents as a painful groin in those sports that involve kicking and twisting movements while running, particularly in rugby, football, soccer, and ice hockey players. Moreover, sportsman's hernia can be encountered even in normally physically active people. The pain experienced is recognized at the common point of origin of the rectus abdominis muscle and the adductor longus tendon on the pubic bone and the insertion of the inguinal ligament on the pubic bone. It is accepted that this chronic pain caused by abdominal wall weakness or injury occurs without a palpable hernia. We proposed the new name "pubic inguinal pain syndrome." In the period between January 2006 and November 2013 all patients afferent in our ambulatory clinic for chronic groin pain without a clinically evident hernia were assessed with medical history, physical examination, dynamic ultrasound, and pelvic and lumbar MRI. All patients were proposed for a conservative treatment and then, if it was not effective, for a surgical treatment. Our etiopathogenetic theory is based on three factors: (1) the compression of the three nerves of the inguinal region, (2) the imbalance in strength of adductor and abdominal wall muscles caused by the hypertrophy and stiffness of the insertion of rectus muscle and adductor longus muscle, and (3) the partial weakness of the posterior wall. Our surgical procedure includes the release of all three nerves of the region, the correction of the imbalance in strength with the partial tenotomy of the rectus and adductor longus muscles, and the repair of the partial weakness of the posterior wall with a lightweight mesh. This treatment reported excellent results with complete relief of symptoms after resumption of physical activity in all cases.


Cytoplasmic MTOCs control spindle orientation for asymmetric cell division in plants.
Proper orientation of the cell division axis is critical for asymmetric cell divisions that underpin cell differentiation. In animals, centrosomes are the dominant microtubule organizing centers (MTOC) and play a pivotal role in axis determination by orienting the mitotic spindle. In land plants that lack centrosomes, a critical role of a microtubular ring structure, the preprophase band (PPB), has been observed in this process; the PPB is required for orienting (before prophase) and guiding (in telophase) the mitotic apparatus. However, plants must possess additional mechanisms to control the division axis, as certain cell types or mutants do not form PPBs. Here, using live imaging of the gametophore of the moss Physcomitrella patens, we identified acentrosomal MTOCs, which we termed "gametosomes," appearing de novo and transiently in the prophase cytoplasm independent of PPB formation. We show that gametosomes are dispensable for spindle formation but required for metaphase spindle orientation. In some cells, gametosomes appeared reminiscent of the bipolar MT "polar cap" structure that forms transiently around the prophase nucleus in angiosperms. Specific disruption of the polar caps in tobacco cells misoriented the metaphase spindles and frequently altered the final division plane, indicating that they are functionally analogous to the gametosomes. These results suggest a broad use of transient MTOC structures as the spindle orientation machinery in plants, compensating for the evolutionary loss of centrosomes, to secure the initial orientation of the spindle in a spatial window that allows subsequent fine-tuning of the division plane axis by the guidance machinery.


High prevalence of "non-dipping" blood pressure and vascular stiffness in HIV-infected South Africans on antiretrovirals.
HIV-infected individuals are at increased risk of tissue inflammation and accelerated vascular aging ('inflamm-aging'). Abnormal diurnal blood pressure (BP) rhythms such as non-dipping may contribute to an increased risk of cardiovascular and cerebrovascular events in HIV infected individuals. However, little data exists on ambulatory blood pressure (ABP) and measures of vascular stiffness in the black African HIV infected population. This is a cross-sectional analysis of otherwise well, HIV infected outpatients on ART for >5 years. Study assessments included: 24hr ABP monitoring, pulse wave velocity (PWV) and central aortic systolic pressure (CASP) using a AtCor Medical Sphygmocor device, fasting lipogram, oral glucose tolerance test, high-sensitivity C-reactive protein (hsCRP) and anthropometric data. Patients completed a questionnaire of autonomic symptoms. CD4+ counts and viral loads were obtained from the National Laboratory results system. Sixty seven black participants were included in the analysis of whom 91% (n = 61) were female with a mean age of 42.2 ± 8.6 years. The median duration on ART was 7.5 years (IQR = 6-10), 84% were virally supressed and the median CD4 count was 529.5cells/mm3 (IQR = 372.0-686.5). The majority (67%) were classified as overweight and 76% had an increased waist circumference, yet only 88% of participants were normotensive. A hsCRP level in the high cardiovascular risk category was found in 68% of participants. The prevalence of non-dipping BP was 65%. Interestingly, there was no association on multivariable analysis between dipping status and traditional risk factors for non-dipping BP, such as: obesity, autonomic dysfunction and older age. This relatively young cross-sectional sample of predominantly normotensive, but overweight black women on effective ART >5 years showed: a high prevalence of non-dipping BP, inflammation and vascular stiffness. Causality cannot be inferred but cardiovascular risk reduction should be emphasized in these patients.


The Chorus of User Voices: Meeting the Challenge of Patient-Reported Outcomes From Medications.
Prescription medication management by both healthcare providers and patients describe struggles not to inflict harm and encourage safe use in the context of culture and social relationships. Risk vs benefit assessments conducted in these contexts are essential for understanding how medication-taking decisions are made. Qualitatively-based Patient Reported Outcomes (PROs) are critical to understanding how medication pathways are navigated by both healthcare providers and patients seeking relief from maladies. Qualitative PROs have the potential to improve understanding of social and cultural factors that impact patient decision making whether to take a drug at all and would also serve to support developing models of patient-centered care.


[Segmented lymph nodes in man].
Segmentary lumbar, posterior pancreato-duodenal and inferior tracheobronchial lymph nodes have been investigated macro- and microscopically. The segmentary lymph nodes reach 12 X 45 X 45 mm, 10 X 25 X 100 mm in size. Most often these nodes are found among the posterior pancreato-duodenal lymph nodes (92%). They represent a conglomerate of smaller lymph nodes growing together and having their own capsule, parenchyma, sinuses, afferent and deferent lymph vessels and are united into one large node.


Conjunctival Lymphoma--An International Multicenter Retrospective Study.
To date, the clinical features of the various subtypes of conjunctival lymphoma (CL) have not been previously evaluated in a large cohort. To characterize subtype-specific clinical features of CL and their effect on patient outcome. A retrospective multicenter study was performed. Patient data were collected from January 1, 1980, through December 31, 2010. The dates of the analysis were May 15, 2015, to August 20, 2015. The median follow-up period was 43 months. Seven eye cancer centers were involved in the study. In total, 268 patients with CL were identified, 5 of whom were excluded because of missing clinical data. Overall survival, disease-specific survival, and progression-free survival were the primary end points. Two hundred sixty-three patients with CL were included in the study. Their mean age was 61.3 years, and 55.1% (145 of 263) were female. All lymphomas were of B-cell type. The most frequent subtype was extranodal marginal zone lymphoma (EMZL) (68.4% [180 of 263]), followed by follicular lymphoma (FL) (16.3% [43 of 263]), mantle cell lymphoma (MCL) (6.8% [18 of 263]), and diffuse large B-cell lymphoma (DLBCL) (4.6% [12 of 263). Conjunctival lymphoma commonly manifested in elderly individuals (age range, 60-70 years old), with EMZL having a female predilection (57.8% [104 of 180]) and MCL having a marked male predominance (77.8% [14 of 18]). Unlike EMZL and FL, DLBCL and MCL were frequently secondary diseases (41.7% [5 of 12] and 88.9% [16 of 18], respectively), with MCL showing a frequent occurrence of stage IVE lymphoma (61.1% [11 of 18]) and bilateral manifestation (77.8% [14 of 18]). Localized disease (stage IE or IIE) was commonly treated with external beam radiation therapy (EBRT) with or without chemotherapy, while widespread lymphoma (stage IIIE or IVE) and MCL of any stage were managed with chemotherapy with or without EBRT. Diffuse large B-cell lymphoma and MCL had a poor prognosis, with 5-year disease-specific survival of 55.0% and 9.0%, respectively, in contrast to EMZL (97.0%) and FL (82.0%). Further survival predictors included age (EMZL), sex (FL), and Ann Arbor staging classification (EMZL and FL). The American Joint Committee on Cancer TNM staging showed limited prognostic usefulness, only being able to predict survival for patients with DLBCL. Conjunctival lymphoma consists of mainly 4 subtypes of B-cell non-Hodgkin lymphoma: EMZL, FL, MCL, and DLBCL. Mantle cell lymphoma is characterized by a particularly high frequency of secondary disease of stage IVE and bilateral manifestation. The histological subtype is the main outcome predictor, with MCL and DLBCL having a markedly poorer prognosis than EMZL and FL.


Respiratory deadspace measurements in neonates during extracorporeal membrane oxygenation.
To evaluate the bias and precision of a simple, bedside method of quantification of minute CO2 production (study 1) and then apply the technique to measure physiologic deadspace in a group of neonates undergoing extracorporeal membrane oxygenation (ECMO) (study 2). A prospective, clinical study comparing a simple method of quantifying minute CO2 production with a criterion standard (study 1); a cohort study evaluating the utility of deadspace measurements in neonates undergoing extracorporeal membrane oxygenation (study 2). Tertiary care pediatric intensive care unit (ICU) in a university hospital. Study 1: Thirteen neonates (weighing from 2.5 to 6.2 kg) were studied during mechanical ventilation being administered for a variety of respiratory diseases. Study 2: Fifteen neonates with respiratory failure were examined during the course of ECMO. INTERVENTIONS (METHODS): Study 1: The CO2 concentration of expired gas sampled at the exhaust port of the test ventilator was continuously measured and transformed to minute CO2 production by the following formula: (calculated minute CO2 production) = (tension of CO2 in exhaust gas) x 0.0013 x (ventilator pneumotachometer minute volume), where 0.0013 is the conversion factor to express gas tension as fractional volume at standard conditions. Minute CO2 production was measured independently with a previously validated bedside calorimeter and the calculated and measured values were compared. Study 2: The CO2 concentration of expired gas sampled at the exhaust port of the test ventilator was continuously measured and transformed to mixed expired CO2 by the following formula, which corrects for compressible volume measured by the ventilator pneumotachometer: mixed expired CO2 = (tension of CO2 in exhaust gas) x (ventilator pneumotachometer minute volume)/(minute volume at proximal airway). We then utilized the Bohr-Enghoff method to calculate the deadspace/tidal volume ratio: deadspace/tidal volume ratio = (PaCO2--mixed expired CO2 tension)/PaCO2. Study 1: Calculated minute CO2 production ranged between 11.8 and 38.9 mL/min. When compared with measured minute CO2 production, the bias and precision were -0.8 and 1.7 mL/min, respectively. The mean percent difference for calculated minute CO2 production was 3.7%. Study 2: Deadspace/tidal volume ratio was calculated during temporary separation from ECMO support as the patients demonstrated improvements in gas exchange and lung compliance. Fifty-two measurements were made in 15 patients, and 13 of 15 patients demonstrated a decrease in deadspace/tidal volume ratio during the course of ECMO. The mean decrease was 21% and the decrease was statistically significant. Minute CO2 production can be measured simply and accurately, using equipment readily available in most ICU settings. The same method can be utilized to calculate the deadspace/tidal volume ratio, which provides valuable information regarding the gas exchanging efficiency of the neonatal lung during ECMO.


Mutations within the kinase domain and truncations of the epidermal growth factor receptor are rare events in bladder cancer: implications for therapy.
It has previously been reported that the patient response to gefitinib depends on the presence of mutations within the kinase domain of epidermal growth factor receptor (EGFR) or the expression of its truncated form, EGFR variant III (EGFRvIII). The focus of this study was to determine if these alterations are present within the tyrosine kinase and ligand-binding domain of EGFR in urothelial carcinoma. The kinase domain found within exons 18 to 21 of the EGFR from 11 bladder cancer cell lines and 75 patient tumors were subjected to automated sequencing. EGFRvIII expression was determined by immunohistochemistry using a urothelial carcinoma tissue microarray, and its expression was subsequently verified by reverse transcription PCR, real-time PCR, and Western blot analysis, using an EGFRvIII-transfected glioblastoma cell line and glioblastoma tumors as positive controls. Our analysis failed to detect mutations within the tyrosine kinase domain of EGFR in the 11 cell lines and 75 patients tested. The initial analysis of EGFRvIII expression by immunohistochemistry revealed that at least 50% of the patient tumors expressed EGFRvIII in a urothelial carcinoma tissue microarray. Conflicting reports exist, however, regarding the extent of EGFRvIII expression in tissues owing to the specificity of the antibodies and the methodologies used. Therefore, we sought to validate this observation by reverse transcription PCR, real-time PCR, and Western blot analysis. In these assays, none of the samples were positive for EGFRvIII except for control transfectants and glioblastomas. When our results are taken together, we conclude that alterations within the tyrosine kinase domain and expression of EGFRvIII are rare events in bladder cancer. The present study has clinical implications in selecting tyrosine kinase inhibitors for the therapy of urothelial carcinoma.


Lack of Durable Improvements in β-Cell Function Following Withdrawal of Pharmacological Interventions in Adults With Impaired Glucose Tolerance or Recently Diagnosed Type 2 Diabetes.
The Restoring Insulin Secretion (RISE) Adult Medication Study compared pharmacological approaches targeted to improve β-cell function in individuals with impaired glucose tolerance (IGT) or treatment-naive type 2 diabetes of <12 months duration. A total of 267 adults with IGT (n = 197, 74%) or recently diagnosed type 2 diabetes (n = 70, 26%) were studied. Participants were randomized to receive 12 months of metformin alone, 3 months of insulin glargine with a target fasting glucose <5 mmol/L followed by 9 months of metformin, 12 months of liraglutide combined with metformin, or 12 months of placebo. β-Cell function was assessed using hyperglycemic clamps at baseline, 12 months (on treatment), and 15 months (3 months off treatment). The primary outcome was β-cell function at 15 months compared with baseline. All three active treatments produced on-treatment reductions in weight and improvements in HbA1c compared with placebo; the greatest reductions were seen in the liraglutide plus metformin group. At 12 months, glucose-stimulated C-peptide responses improved in the three active treatment groups and were greatest in the liraglutide plus metformin group, but the arginine-stimulated incremental C-peptide response was reduced in the liraglutide plus metformin group. Despite on-treatment benefits, 3 months after treatment withdrawal there were no sustained improvements in β-cell function in any treatment group. In adults with IGT or recently diagnosed type 2 diabetes, interventions that improved β-cell function during active treatment failed to produce persistent benefits after treatment withdrawal. These observations suggest that continued intervention may be required to alter the progressive β-cell dysfunction in IGT or early type 2 diabetes.


Meeting the demand for innate and adaptive immunities during evolution.
An ideal immune system should provide each individual with rapid and efficient responses, a diverse repertoire of recognition and effector molecules and a certain flexibility to match the changing internal and external environment. It should be economic in cells and genes. Specific memory would be useful. It should not be autoreactive. These requirements, a mixture of innate and adaptive immunity features, are modulated in function of the dominant mode of selection for each species of metazoa during evolution (K or r). From sponges to man, a great diversity of receptors and effector mechanisms, some of them shared with plants, are articulated around conserved signalling cascades. Multiple attempts at combining innate and adaptive immunity somatic features can be observed as new somatic mechanisms provide individualized repertoires of receptors throughout metazoa, in agnathans, prochordates, echinoderms and mollusks. The adaptive immunity of vertebrates with lymphocytes and their specific receptors of the immunoglobulin superfamily, the major histocompatibility complex, developed from innate immunity evolutionary lines that can be traced back in earlier deuterostomes.


Bacterial vaginosis - More questions than answers.
Bacterial vaginosis is the commonest cause of abnormal vaginal discharge in women of reproductive age and is associated with serious pregnancy related sequelae and increased transmission of sexually transmissible infections, including HIV. The aetiology, pathology, microbiology and transmission of bacterial vaginosis remain poorly understood. This article discusses the prevalence, clinical features and possible complications of bacterial vaginosis. It summarises what is known about the aetiology, pathophysiology and treatment of the condition and highlights directions for further research. Bacterial vaginosis is characterised by a complex disturbance of the normal vaginal flora with an overgrowth of anaerobic and other micro-organisms and a corresponding decrease in important lactobacillus species. The cause is not known, but observational evidence suggests the possibility of sexual transmission. Bacterial vaginosis is diagnosed by the Amsel or the Nugent method. Recommended treatment is with 7 days of oral metronidazole or vaginal clindamycin. More than 50% of women will experience recurrence of bacterial vaginosis within 6 months. It is not known whether this represents relapse or re-infection. Further research is needed into the aetiology, pathogenesis and optimal treatment of this condition.


Human D-Amino Acid Oxidase: Structure, Function, and Regulation.
D-Amino acid oxidase (DAAO) is an FAD-containing flavoenzyme that catalyzes with absolute stereoselectivity the oxidative deamination of all natural D-amino acids, the only exception being the acidic ones. This flavoenzyme plays different roles during evolution and in different tissues in humans. Its three-dimensional structure is well conserved during evolution: minute changes are responsible for the functional differences between enzymes from microorganism sources and those from humans. In recent years several investigations focused on human DAAO, mainly because of its role in degrading the neuromodulator D-serine in the central nervous system. D-Serine is the main coagonist of N-methyl D-aspartate receptors, i.e., excitatory amino acid receptors critically involved in main brain functions and pathologic conditions. Human DAAO possesses a weak interaction with the FAD cofactor; thus, in vivo it should be largely present in the inactive, apoprotein form. Binding of active-site ligands and the substrate stabilizes flavin binding, thus pushing the acquisition of catalytic competence. Interestingly, the kinetic efficiency of the enzyme on D-serine is very low. Human DAAO interacts with various proteins, in this way modulating its activity, targeting, and cell stability. The known properties of human DAAO suggest that its activity must be finely tuned to fulfill a main physiological function such as the control of D-serine levels in the brain. At present, studies are focusing on the epigenetic modulation of human DAAO expression and the role of post-translational modifications on its main biochemical properties at the cellular level.


Cerebral aspergillosis within new tumour site presents as incidental new brain lesion in patient receiving temozolomide for glioblastoma multiforme.
Glioblastoma multiforme (GBM) is an aggressive tumour that can lead to lymphopaenia. Its standard treatment involves temozolomide (TMZ) chemotherapy with radiation, often with addition of corticosteroids for symptomatic management. Although TMZ is also immunosuppressive, patients receiving TMZ rarely develop disseminated opportunistic infections. Here, we report the case of a patient with GBM receiving TMZ, radiotherapy and corticosteroids, who develops an incidental new brain lesion that is found to be disseminated Aspergillus within a new GBM tumour site. The patient received successful early treatment of her central nervous system aspergillosis. This case illustrates the profound immunosuppressive potential of GBM in conjunction with TMZ and corticosteroids, which can lead to high-morbidity opportunistic infections concurrently with tumour progression. Future research is needed to elucidate GBM, TMZ and corticosteroids' compound immune effects and guide management that strikes a balance between treating high-morbidity infections and continuing with immunosuppressive chemotherapy.


Antibiotics in the Biliary Tract: A Review of the Pharmacokinetics and Clinical Outcomes of Antibiotics Penetrating the Bile and Gallbladder Wall.
Biliary tract infections (BTIs), including cholangitis and cholecystitis, are common causes of bacteremia. Bacteremic BTIs are associated with a mortality rate of 9-12%. The extent to which antibiotics are excreted in the bile and the ratio of their exposure to the minimum inhibitory concentration of the infecting organism are among the important factors for the treatment of BTIs. This review updates health care professionals on the distribution of antibiotics in the common bile duct, gallbladder, and gallbladder wall. Antibiotic efficacy in treating BTIs based on the latest available clinical studies is also discussed. The efficacy and pharmacokinetics of 50 antibiotics are discussed. Overall, most antibiotic classes exhibit biliary penetration that translates into clinical efficacy. Only seven antibiotics (amoxicillin, cefadroxil, cefoxitin, ertapenem, gentamicin, amikacin, and trimethoprim/sulfamethoxazole) had poor biliary penetration profiles. Three antibiotics (ceftibuten, ceftolozane/tazobactam, and doripenem) had positive clinical outcomes despite the lack of pharmacokinetic studies on their penetration into the biliary tract. Conflicting efficacy data were reported for ampicillin despite adequate biliary penetration, whereas conflicting pharmacokinetic data were reported with cefaclor and moxifloxacin. Even in the absence of supportive clinical studies, antibiotics with good biliary penetration profiles may have a place in the treatment of BTIs.


Translation to Primary Care of an Effective Teen Safe Driving Program for Parents.
Addressing teen driver crashes, this study adapted an effective Checkpoints(TM) program for parents of teen drivers for dissemination by primary care practitioners (PCPs) and the web; distributed the PCP/web program through pediatric practices; and examined dissemination to/implementation by parents. The website, youngDRIVERparenting.org, and brief intervention protocol were developed. PCPs delivered interventions and materials to parents, referred them to the website, and completed follow-up surveys. Google Analytics assessed parents' website use. Most PCPs reported delivering interventions with fidelity, and thought the program important and feasible. Brief interventions/website referrals, averaging 4.4 minutes, were delivered to 3465 (87%) of 3990 eligible parents by 133 PCPs over an 18-week average. Website visits (1453) were made by 42% of parents, who spent on average 3:53 minutes viewing 4.2 topics. This program costs little (its website, training and promotional materials are available) and could be one component of a comprehensive approach to reducing teen driver crashes.


Evaluation of left ventricular function in patients with coronary slow flow: A systematic review and meta-analysis.
Coronary slow flow (CSF) is an angiographic finding defined as delayed distal vessel perfusion without severe stenosis of the epicardial coronary arteries. However, definite alterations in left ventricular (LV) function in patients with CSF remains inconsistent. This study aimed to clarify the changes in LV function in patients with CSF and explore the factors that may influence LV function. PubMed, Embase, and Cochrane Library databases were systematically searched. Standardized mean differences and 95% confidence intervals (CI) for the LV function parameters were calculated. Subgroup analysis, meta-regression analysis, and correlation analysis were performed to explore the factors influencing LV function. Twenty-two studies (1101 patients with CSF) were included after searching three databases. In patients with CSF, LV ejection function in patients with CSF was marginally lower (61.8%; 95% CI: 61.0%, 62.7%), global longitudinal strain was decreased (-18.2%; 95% CI: -16.7%, -19.7%). Furthermore, left atrial diameter, left atrial volume index, and E/e' were significantly increased, while E/A and e' were significantly decreased. The mean thrombolysis in myocardial infarction frame count (TFC) was linearly associated with LV function; the larger the mean TFC, the greater the impairment of LV function. Left ventricular systolic and diastolic functions were impaired in patients with CSF, and this impairment was aggravated with increasing mean TFC.


Gastric dilatation and volvulus in a brachycephalic dog with hiatal hernia.
A brachycephalic dog was presented with an acute onset of retching and abdominal discomfort. The dog had a chronic history of stertor and exercise intolerance suggestive of brachycephalic airway obstructive syndrome. Radiographs were consistent with a Type II hiatal hernia. The dog was referred and within hours of admission became acutely painful and developed tympanic abdominal distension. A right lateral abdominal radiograph confirmed gastric dilatation and volvulus with herniation of the pylorus through the hiatus. An emergency exploratory coeliotomy was performed, during which the stomach was derotated, and an incisional gastropexy, herniorrhaphy and splenectomy were performed. A staphylectomy was performed immediately following the exploratory coeliotomy. The dog recovered uneventfully. Gastric dilatation and volvulus is a potentially life-threatening complication that can occur in dogs with Type II hiatal hernia and should be considered a surgical emergency.


Isolation and characterization of a tobacco mosaic virus-inducible myb oncogene homolog from tobacco.
Salicylic acid (SA) plays an important role in signaling the activation of plant defense responses against pathogen attack including induction of pathogenesis-related (PR) proteins. To gain further insight into the SA-mediated signal transduction pathway, we have isolated and characterized a tobacco mosaic virus (TMV)-inducible myb oncogene homolog (myb1) from tobacco. The myb1 gene was induced upon TMV infection during both the hypersensitive response and development of systemic acquired resistance in the resistant tobacco cultivar following the rise of endogenous SA, but was not activated in the susceptible cultivar that fails to accumulate SA. The myb1 gene was also induced by incompatible bacterial pathogen Pseudomonas syringae pv. syringae during the hypersensitive response. Exogenous SA treatment rapidly (within 15 min) activated the expression of myb1 in both resistant and susceptible tobacco cultivars with the subsequent induction of PR genes occurring several hours later. Biologically active analogs of SA and 2,6-dichloroisonicotinic acid (a synthetic functional analog of SA), which induce PR genes and enhanced resistance, also activated the myb1 gene. In contrast, biologically inactive analogs were poor inducers of myb1 gene expression. Furthermore, the recombinant Myb1 protein was shown to specifically bind to a Myb-binding consensus sequence found in the promoter of the PR-1a gene. Taken together, these results suggest that the tobacco myb1 gene encodes a signaling component down-stream of SA that may participate in transcriptional activation of PR genes and plant disease resistance.


Bovine milk-derived exosomes from colostrum are enriched with proteins implicated in immune response and growth.
Exosomes are extracellular vesicles secreted by multiple cell types into the extracellular space. They contain cell-state specific cargos which often reflects the (patho)physiological condition of the cells/organism. Milk contains high amounts of exosomes and it is unclear whether their cargo is altered based on the lactation stage of the organism. Here, we isolated exosomes from bovine milk that were obtained at various stages of lactation and examined the content by quantitative proteomics. Exosomes were isolated by OptiPrep density gradient centrifugation from milk obtained from cow after 24, 48 and 72 h post calving. As control, exosomes were also isolated from cows during mid-lactation period which has been referred to as mature milk (MM). Biochemical and biophysical characterization of exosomes revealed the high abundance of exosomes in colostrum and MM samples. Quantitative proteomics analysis highlighted the change in the proteomic cargo of exosomes based on the lactation state of the cow. Functional enrichment analysis revealed that exosomes from colostrum are significantly enriched with proteins that can potentially regulate the immune response and growth. This study highlights the importance of exosomes in colostrum and hence opens up new avenues to exploit these vesicles in the regulation of the immune response and growth.


Incidence of Japanese encephalitis among acute encephalitis syndrome cases in West Bengal, India.
Japanese encephalitis (JE) is the most important cause of acute and epidemic viral encephalitis. Every year sporadic JE cases are reported from the various districts of West Bengal, indicating its endemicity in this state. JE vaccination programme has been undertaken by the State Health Department of West Bengal. This study was aimed at seeing the present scenario of JE among acute encephalitis syndrome (AES) cases in West Bengal. Blood and/or CSF samples were referred from suspected AES cases to the referral virology laboratory of the Calcutta School of Tropical Medicine from different hospitals of Kolkata. IgM antibody capture ELISA was performed on the CSF and serum samples by JE virus MAC ELISA kit supplied by the National Institute of Virology, Pune. The present study reveals that 22.76% and 5% of the AES cases were positive for JE IgM in 2011 and 2012, respectively. JE is mainly prevalent in children and adolescents below 20 years of age with no gender predilection. Although the percentages of JE positive cases were high in 2011, it sharply decreased thereafter possibly due to better awareness programs, due to mass vaccination, or simply due to natural epidemiological niche periodicity due to herd immunity.


Reflection of passive exposure to smoking in the home on the prevalence of chronic bronchitis in non-smokers.
Analysis of data assembled in an epidemiological investigation of 123 235 adults from three districts of the West Bohemian region incl. 57 217 male non-smokers from a total of 66 806 and 23 473 male non-smokers from a total number of 56 429 males revealed a high significant (p less than 0.005) rise in the prevalence of symptoms of chronic bronchitis in subjects exposed passively to smoking in the home. Exposure to other adverse factors increased the prevalence approximately three times in both sexes. The effect of passive exposure to smoking was, however, manifested far more in men. No reason was found in the investigated data for the interpretation of these differences. It appears that the cause could be the different behaviour of smokers to non-smokers in the home, where the smoker-husband usually is considerate to his non-smoker wife during her pregnancy and child care, while in women smokers this motivation is lacking.


[Molecular structure of the locus for sucrose utilization by Lactobacillus plantarum: comparison with Pediococcus pentosaceus].
Structure of the sucrose utilization locus in a Lactobacillus plantarum type strain was studied using PCR and Southern hybridization. Restriction map analysis revealed its high similarity to the sequenced sucrose utilization locus of Pediococcus pentosaceus pSRQ1. The L. plantarum locus proved containing oppositely oriented scrA and the scrBRagl operon, but not agaS. The L. plantarum sucrase gene (scrB) was partly sequenced. A higher (98.6%) homology was revealed between scrB than between the 16S rRNA genes of L. plantarum and P. pentosaceus, suggesting horizontal transfer of the sucrose utilization locus between the genera of lactic acid bacteria. Amino acid sequence analysis showed that the ScrB proteins of the two species belong to a subfamily of glycosyl hydrolase family GH32 which includes various beta-fructosidases.


1,2-benzisoxazole phosphorodiamidates as novel anticancer prodrugs requiring bioreductive activation.
Several 1,2-benzisoxazole phosphorodiamidates have been designed as prodrugs of phosphoramide mustard requiring bioreductive activation. Enzymatic reduction of 1,2-benziosoxazole moiety is expected to result in the formation of imine intermediate due to the cleavage of the N-O bond. The imine should then be spontaneously hydrolyzed to a ketone metabolite, thereby facilitating base-catalyzed beta-elimination of cytotoxic phosphoramide mustard. As expected, the proposed prodrugs 4, 9, and 12 were at least 3-5-fold more potent cytotoxins than control compounds 5 and 15, which lack in the phosphoramide mustard group. Upon incubation with phenobarb-induced rat liver S-9 fraction, compounds 4, 9, and 12 underwent extensive NADPH-dependent metabolism with concomitant generation of alkylating activity under both hypoxic and oxic conditions. Corresponding ketone metabolites were detected for 9 and 15. NADPH-dependent bioreduction of 15 to its ketone metabolite 16 was located in the microsomal fraction and inhibited by SKF-525A and pCMBA. Compared with phenobarb-induced rat liver microsomal fraction, incubation of 15 with rat or human p450 reductase microsomes showed moderate generation of 16. Microsomal cytochrome p450 and/or p450 reductase appear to be involved in the reductive metabolism of 1,2-benzisoxazole moiety under hypoxic as well as oxic conditions.


Computational modeling of tumor response to vascular-targeting therapies--part I: validation.
Mathematical modeling techniques have been widely employed to understand how cancer grows, and, more recently, such approaches have been used to understand how cancer can be controlled. In this manuscript, a previously validated hybrid cellular automaton model of tumor growth in a vascularized environment is used to study the antitumor activity of several vascular-targeting compounds of known efficacy. In particular, this model is used to test the antitumor activity of a clinically used angiogenesis inhibitor (both in isolation, and with a cytotoxic chemotherapeutic) and a vascular disrupting agent currently undergoing clinical trial testing. I demonstrate that the mathematical model can make predictions in agreement with preclinical/clinical data and can also be used to gain more insight into these treatment protocols. The results presented herein suggest that vascular-targeting agents, as currently administered, cannot lead to cancer eradication, although a highly efficacious agent may lead to long-term cancer control.


Conjugation of benzylvanillin and benzimidazole structure improves DNA binding with enhanced antileukemic properties.
Benzyl-o-vanillin and benzimidazole nucleus serve as important pharmacophore in drug discovery. The benzyl vanillin (2-(benzyloxy)-3-methoxybenzaldehyde) compound shows anti-proliferative activity in HL60 leukemia cancer cells and can effect cell cycle progression at G2/M phase. Its apoptosis activity was due to disruption of mitochondrial functioning. In this study, we have studied a series of compounds consisting of benzyl vanillin and benzimidazole structures. We hypothesize that by fusing these two structures we can produce compounds that have better anticancer activity with improved specificity particularly towards the leukemia cell line. Here we explored the anticancer activity of three compounds namely 2-(2-benzyloxy-3-methoxyphenyl)-1H-benzimidazole, 2MP, N-1-(2-benzyloxy-3-methoxybenzyl)-2-(2-benzyloxy-3-methoxyphenyl)-1H-benzimidazole, 2XP, and (R) and (S)-1-(2-benzyloxy-3-methoxyphenyl)-2, 2, 2-trichloroethyl benzenesulfonate, 3BS and compared their activity to 2-benzyloxy-3-methoxybenzaldehyde, (Bn1), the parent compound. 2XP and 3BS induces cell death of U937 leukemic cell line through DNA fragmentation that lead to the intrinsic caspase 9 activation. DNA binding study primarily by the equilibrium binding titration assay followed by the Viscosity study reveal the DNA binding through groove region with intrinsic binding constant 7.39 µM/bp and 6.86 µM/bp for 3BS and 2XP respectively. 2XP and 3BS showed strong DNA binding activity by the UV titration method with the computational drug modeling showed that both 2XP and 3BS failed to form any electrostatic linkages except via hydrophobic interaction through the minor groove region of the nucleic acid. The benzylvanillin alone (Bn1) has weak anticancer activity even after it was combined with the benzimidazole (2MP), but after addition of another benzylvanillin structure (2XP), stronger activity was observed. Also, the combination of benzylvanillin with benzenesulfonate (3BS) significantly improved the anticancer activity of Bn1. The present study provides a new insight of benzyl vanillin derivatives as potential anti-leukemic agent.


Altered cleavage site preference of a proteolytic antibody light chain induced by denaturation.
A recombinant antibody light chain (L chain) maintained under non-denaturing conditions displayed preferential cleavage of synthetic peptides conjugated to methylcoumarinamide (MCA) on the C-terminal side of Arg and Lys residues. The same L chain renatured from a denaturing solvent (guanidine hydrochloride) acquired the capability of cleaving Tyr-MCA and Leu-MCA bonds, and its ability to cleave MCA linked to basic residues was decreased. The altered cleavage preference was accompanied by a conformational transition in the protein, evident from the fluorescence emission spectra. These observations suggest the feasibility of redirecting the cleavage specificity via alterations in the conformation of proteolytic antibody combining sites.


Interobserver variability in the diagnosis of crypt dysplasia in Barrett esophagus.
Recent morphologic and molecular evidence suggests that dysplasia in Barrett esophagus (BE) begins in the bases of the crypts [crypt dysplasia (CD)] and progresses with time to involve the upper portions of the crypts and surface epithelium. The aim of this study was to evaluate the criteria and reproducibility of diagnosing CD among 6 gastrointestinal pathologists, all with research interest in BE. Six gastrointestinal pathologists evaluated 2 clinical study sets, the first consisting of 40 BE cases [BE: 10, CD: 9, low-grade dysplasia (LGD): 10, high-grade dysplasia (HGD); 9, and intramucosal adenocarcinoma; 2] and the second consisting of 63 cases (BE: 16, CD: 15, LGD: 15, HGD: 15, and intramucosal adenocarcinoma: 2), at least 4 months apart. In between evaluations, all of the pathologists met at 1 hospital (consensus conference) to review the areas of disagreement and establish more objective criteria. Overall, the level of agreement for all cases was moderate (κ=0.44), and the level of agreement did not change significantly after evaluation of the second study set. The highest levels of agreement were obtained for lesions at the low and high end of the spectrum (BE without dysplasia and HGD). Overall, the degree of agreement for CD was moderate after both the first and second study set review (κ=0.44 and 0.46, respectively). However, the degree of agreement for CD was higher than that obtained for LGD in both study sets. In the first study set, 4 or more pathologists agreed with the original CD diagnosis in 78% of cases, and this value did not change significantly after review of the second study set. The observers agreed that characteristic features of CD include the presence of unequivocal dysplastic cells, similar in appearance to traditional LGD, involving any part, or all of the length, of the crypt in the absence of intercrypt surface epithelial involvement. Rare cases of CD may show high-grade cytologic features composed of markedly enlarged nuclei with increased nuclear/cytoplasmic ratio, eosinophilic cytoplasm, irregular nuclear membranes, and loss of polarity. The findings in this study suggest that CD can be diagnosed reliably with a moderate level of interobserver agreement. Long-term and multi-institutional studies should be carried out to further determine the biological and clinical significance and natural history of CD in patients with BE.


Regulation of hepatic acute phase protein synthesis by products of interleukin 2 (IL 2)-stimulated human peripheral blood mononuclear cells.
Cancer patients injected with recombinant human IL 2 develop marked changes in serum concentrations of hepatic acute-phase proteins. To determine if this acute-phase response involves a change in the rate of hepatic protein synthesis and if it is due to a direct effect of IL 2 on hepatocytes, human hepatoma-derived hepatocytes (Hep-3B cells) were incubated in medium containing IL 2 or in culture supernatants from IL 2-activated human peripheral blood mononuclear cells (PBMNC). The rate of synthesis of two acute-phase proteins, complement protein factor B and albumin, was determined by the incorporation of a radiolabeled amino acid precursor into newly synthesized protein as measured by analytical gel electrophoresis of immunoprecipitates. IL 2 in concentrations from 1 to 1000 U/ml had no effect on the synthesis of factor B or albumin; conversely, there was a dose-dependent increase in the rate of synthesis of factor B and decrease in albumin synthesis mediated by culture supernatants of IL 2-activated PBMNC. The magnitude of the effect of acute-phase protein synthesis was dependent on the IL 2 concentration used for the activation of PBMNC. The rate of factor B synthesis increased approximately 4.0-fold in the presence of culture supernatants of PBMNC activated with either opsonized heat-killed Staphylococcus albus or with 1000 U/ml IL 2. Preincubation of the IL 2-activated PBMNC culture supernatants with an antiserum specific for recombinant IL 1-beta completely neutralized the capacity of the supernatants to stimulate factor B synthesis, whereas antisera specific for human IL 1-alpha or for tumor necrosis factor had no effect. These results indicate that the indirect effect of IL 2 on hepatic acute phase protein synthesis is mediated by IL 1-beta.


The association between ocular surface measurements with visual field reliability indices and gaze tracking results in preperimetric glaucoma.
To investigate the relationship between gaze tracking (GT) results and ocular surface condition in glaucoma. The Humphrey 24-2 visual field (VF) was measured in 34 eyes of 30 patients with open-angle glaucoma without VF damage. Tear break-up time, Schirmer's test, tear meniscus volume (TMV) and presence of superficial punctate keratopathy (SPK) were also measured in order to describe the condition of the ocular surface. Various GT parameters were calculated: the average frequency of eye movements per stimulus between 1° and 2° (move1-2), the average frequency of eye movements per stimulus between 3° and 5° (move3-5), the average frequency of eye movements per stimulus more than 6° (move≥6), the average tracking failure frequency per stimulus (TFF) and the average blinking frequency. The relationship between GT parameters, reliability indices and ocular surface measurements was investigated using linear mixed modelling. SPK was positively associated with high rates of move3-5 (coefficient=0.12 for SPK+, p=0.003) and move≥6 (coefficient=0.052 for SPK+, p=0.023). High TMV was significantly related to TFF (coefficient=0.37, p=0.023). Fixation losses, false-positives and false-negatives were not significantly associated with any GT parameters or ocular surface measurements. SPK is associated with increased frequency of eye movements (move3-5 and move≥6). In addition, large TMV is associated with increased rate of TFF. Careful attention should be paid when interpreting GT parameters in patients with SPK or a large TMV.


Total therapy with tandem transplants for newly diagnosed multiple myeloma.
Between August 1990 and August 1995, 231 patients (median age 51, 53% Durie-Salmon stage III, median serum beta-2-microglobulin 3.1 g/L, median C-reactive protein 4 g/L) with symptomatic multiple myeloma were enrolled in a program that used a series of induction regimens and two cycles of high-dose therapy ("Total Therapy"). Remission induction utilized non-cross-resistant regimens (vincristine-doxorubicin-dexamethasone [VAD], high-dose cyclophosphamide and granulocyte-macrophage colony-stimulating factor with peripheral blood stem cell collection, and etoposide-dexamethasone-cytarabine-cisplatin). The first high-dose treatment comprised melphalan 200 mg/m2 and was repeated if complete (CR) or partial (PR) remission was maintained after the first transplant; in case of less than PR, total body irradiation or cyclophosphamide was added. Interferon--2b maintenance was used after the second autotransplant. Fourteen patients with HLA-compatible donors underwent an allograft as their second high-dose therapy cycle. Eighty-eight percent completed induction therapy whereas first and second transplants were performed in 84% and 71% (the majority within 8 and 15 months, respectively). Eight patients (3%) died of toxicity during induction, and 2 (1%) and 6 (4%) during the two transplants. True CR and at least a PR (PR plus CR) were obtained in 5% (34%) after VAD, 15% (65%) at the end of induction, and 26% (75%) after the first and 41% (83%) after the second transplants (intent-to-treat). Median overall (OS) and event-free (EFS) survival durations were 68 and 43 months, respectively. Actuarial 5-year OS and EFS rates were 58% and 42%, respectively. The median time to disease progression or relapse was 52 months. Among the 94 patients achieving CR, the median CR duration was 50 months. On multivariate analysis, superior EFS and OS were observed in the absence of unfavorable karyotypes (11q breakpoint abnormalities, -13 or 13-q) and with low beta-2-microglobulin at diagnosis. CR duration was significantly longer with early onset of CR and favorable karyotypes. Time-dependent covariate analysis suggested that timely application of a second transplant extended both EFS and OS significantly, independent of cytogenetics and beta-2-microglobulin. Total Therapy represents a comprehensive treatment approach for newly diagnosed myeloma patients, using multi-regimen induction and tandem transplantation followed by interferon maintenance. As a result, the proportion of patients attaining CR increased progressively with continuing therapy. This observation is particularly important because CR is a sine qua non for long-term disease control and, eventually, cure.


How Do Individuals Develop Alcohol Use Disorder After Bariatric Surgery? A Grounded Theory Exploration.
Bariatric surgery is the most effective treatment for severe obesity. However, following Roux-en-Y gastric bypass (RYGB) surgery, a small minority of patients develop new-onset alcohol use disorder (AUD), the aetiology of which is poorly understood. The aim is to construct a theory to explain the development of AUD among a sample of individuals who reported problematic drinking following RYGB. Semi-structured interviews were conducted with eight RYGB patients diagnosed with AUD attending a multi-disciplinary outpatient weight management service at a public hospital in the Republic of Ireland. A constructivist grounded theory methodology was used to analyse interview transcripts. Participants' main concern was identified as 'unresolved psychological issues' which were managed by 'external coping mechanisms', namely, 'eating to cope'. After RYGB, comfort eating was no longer possible to the same extent. Following a 'honeymoon period', participants' need for an external coping mechanism resurfaced. 'Filling the void' provides a framework to explain how participants managed the symptoms of their unresolved psychological issues through 'behavioural substitution', that is, drinking alcohol instead of eating. The theoretical framework of 'filling the void' adds to contemporary research that conceptualises AUD behavioural substitution as 'addiction transfer' by describing the process by which the phenomenon occurs as well as the characteristics of participants. The clinical implication of this research is to advocate for a reshaping of treatment of RYGB patients, with increased psychological input following surgery.


Changes in erythropoiesis and renal ultrastructure during exposure of mice to hypoxia.
During hypoxia, elevated ESF levels occur which are accompanied by increased erythropoietic activity resulting in progressive elevation of the haemoglobin and haematocrit concentration. An attempt has been made to correlate ultrastructural changes in the kidney with changes in serum ESF concentrations and the erythropoietic state of mice exposed to continuous hypoxia for 3 weeks. A series of changes in the epitheloid and proximal tubules cells occurred which may be related to the biogenesis of erythropoietin. However, no detectable ESF or renal erythropoietic factor could be extracted from the kidneys at the time of maximal ultrastructural changes. Although some ESF-inhibitory material was demonstrated, it could not be extracted into lipid solvents. It is suggested that the failure to consistently extract an erythropoietic factor from the kidneys of hypoxic mice may be related to the absence of renal storage at the onset of increased peripheral demand. Other levels and durations of hypoxia may help to elucidate the role of the kidneys in regulating serum ESF levels.


The Sports-Related Injuries and Illnesses in Paralympic Sport Study (SRIIPSS): a study protocol for a prospective longitudinal study.
Paralympic sport provides sporting opportunities for athletes with a disability, with the Paralympic Games as the main event. Participation in sport is, however, associated with a significant risk for sustaining injuries and illnesses. Our knowledge of sports-related injuries and illnesses in Paralympic sport is very limited and there are no large-scale epidemiological cohort studies. The purpose here is to present a protocol for a prospective longitudinal study: The Sports-Related Injuries and Illnesses in Paralympic Sport Study (SRIIPSS). An argument-based method for investigation of design problems was used to structure the study protocol. The primary requirement of the protocol is to allow prospective studies over time and include exposure to both training and competition. To reflect the complexity of Paralympic sport with athletes' pre-existing impairments, use of assistive equipment, pain and other and medical issues, it is required that the data collection system is specifically adapted to Paralympic sport. To allow the collection of data, at the same time as there is limited access to coaches and medical personnel, it is advantageous that data can be collected online directly from the athletes. Based on this a self-report athlete monitoring system will be developed, where the athletes can enter data weekly via their mobile phones or lap-tops. Data will be collected from around 100 Swedish Paralympic athletes for approximately 1 year, which will allow us to i) prospectively estimate the annual incidence of sports-related injuries and illnesses and ii) explore risk factors and mechanisms for sustaining sports-related injuries and illnesses based on athlete exposure and training loads. For effective implementation of injury and illness prevention measures, comprehensive epidemiological knowledge is required. This study will be the first prospective longitudinal self-report study of sports-related injuries and illnesses in Paralympic sport over a longer period of time. The results will eventually contribute to the development of evidence-based preventive measures specifically adapted to Paralympic sport in order to provide safe and healthy sport participation. Thereby, the project will be of relevance for Paralympic athletes at all levels and to the Paralympic Movement. The study is registered at ClinicalTrials.gov (Identifier: NCT02788500; Registration date: 22 May 2016).


Caffeine ingestion acutely enhances muscular strength and power but not muscular endurance in resistance-trained men.
The goal of this randomized, double-blind, cross-over study was to assess the acute effects of caffeine ingestion on muscular strength and power, muscular endurance, rate of perceived exertion (RPE), and pain perception (PP) in resistance-trained men. Seventeen volunteers (mean ± SD: age = 26 ± 6 years, stature = 182 ± 9 cm, body mass = 84 ± 9 kg, resistance training experience = 7 ± 3 years) consumed placebo or 6 mg kg-1 of anhydrous caffeine 1 h before testing. Muscular power was assessed with seated medicine ball throw and vertical jump exercises, muscular strength with one-repetition maximum (1RM) barbell back squat and bench press exercises, and muscular endurance with repetitions of back squat and bench press exercises (load corresponding to 60% of 1RM) to momentary muscular failure. RPE and PP were assessed immediately after the completion of the back squat and bench press exercises. Compared to placebo, caffeine intake enhanced 1RM back squat performance (+2.8%; effect size [ES] = 0.19; p = .016), which was accompanied by a reduced RPE (+7%; ES = 0.53; p = .037), and seated medicine ball throw performance (+4.3%, ES = 0.32; p = .009). Improvements in 1RM bench press were not noted although there were significant (p = .029) decreases in PP related to this exercise when participants ingested caffeine. The results point to an acute benefit of caffeine intake in enhancing lower-body strength, likely due to a decrease in RPE; upper-, but not lower-body power; and no effects on muscular endurance, in resistance-trained men. Individuals competing in events in which strength and power are important performance-related factors may consider taking 6 mg kg-1 of caffeine pre-training/competition for performance enhancement.


Long-term follow-up of patients receiving injection therapy for erectile dysfunction.
During the last decade, vasoactive intracavernosal pharmacotherapy (VIP) has been used extensively for the treatment of erectile dysfunction. However, there is concern about high discontinuation rates and the possibility of long-term complications. Because of few long-term studies on VIP, we investigated efficacy, side effects, satisfaction index, and drop-out rate for injection therapy in patients who started treatment more than 5 years ago. Questionnaires were mailed to 108 patients who were started on VIP more than 5 years ago, between November 1984 and July 1989. The hospital records and data from the 100 responders (93%) were reviewed. Only 32% of the patients continue to use VIP. Most (56%) of those who discontinued did so during the first year. The patients cited one or more of the following reasons for discontinuation: desire for a permanent modality of therapy (29%), lack of a suitable partner (26%), fear of needles (23%), poor response (23%), fear of complications (22%), and lack of sexual spontaneity (21%). This study, which has one of the longest follow-up periods in the literature, has significant new findings in three areas: discontinuation rates fall after 2 years, long-term complications are relatively minor, and patients who discontinue therapy are significantly older or have a poor initial impression of VIP. Paradoxically, discontinuing VIP was apparently unrelated to side effects or etiology of erectile dysfunction, and 82% of patients would still recommend VIP to a friend. This study conclusively shows that because of high initial satisfaction and relatively minor side effects, VIP should remain as one of the initial options for long-term treatment of erectile dysfunction. However, despite seemingly doing well, patients often discontinue therapy, and therefore should be followed closely so that alternative therapy can be offered.


A spectroscopic approach to imaging and quantification of cartilage lesions in human knee joints.
We have previously described a technology based on diffuse reflectance of broadband light for measuring joint articular cartilage thickness, utilizing that optical absorption is different in cartilage and subchondral bone. This study is the first evaluation of the technology in human material. We also investigated the prospects of cartilage lesion imaging, with the specific aim of arthroscopic integration. Cartilage thickness was studied ex vivo in a number of sites (n = 87) on human knee joint condyles, removed from nine patients during total knee replacement surgery. A reflectance spectrum was taken at each site and the cartilage thickness was estimated using the blue, green, red and near-infrared regions of the spectrum, respectively. Estimated values were compared with reference cartilage thickness values (taken after sample slicing) using an exponential model. Two-dimensional Monte Carlo simulations were performed in a theoretical analysis of the experimental results. The reference cartilage thickness of the investigated sites was 1.60 ± 1.30 mm (mean ± SD) in the range 0-4.2 mm. Highest correlation coefficients were seen for the calculations based on the near-infrared region after normalization to the red region (r = 0.86) and for the green region (r = 0.80).


Cerebellar susceptibility to experimental autoimmune encephalomyelitis in SJL/J mice: potential interaction of immunology with vascular anatomy.
Experimental autoimmune encephalomyelitis (EAE) is utilized as an animal model for multiple sclerosis (MS). In both EAE and MS, activated T-lymphocytes specific for self-antigens present in myelin are linked to CNS inflammation and the breakdown of the blood-brain barrier (BBB) to peripheral blood leukocytes and plasma proteins, predominately in myelin rich white matter. One aspect of MS that has received relatively little attention is the finding that certain CNS regions are more likely than others to develop disease in different patient populations. Understanding the factors predisposing specific brain regions to autoimmune attack, or protecting other regions, would provide a better understanding of the disease as a process, and may also offer additional targets for therapeutic development. EAE offers a model to search for these factors and the first step in such a process is to identify the brain regions that are susceptible to EAE. Until recently the spinal cord in rodents has been considered the region most susceptible to EAE, with disease in more rostral regions occurring later and with reduced severity. However a more recent study has shown that the cerebellum of SJL/J mice, like the spinal cord, is especially susceptible to BBB breakdown in EAE. Although many factors known to be involved in BBB formation and breakdown remain to be assessed for their possible role in increasing the susceptibility of the cerebellum, one potentially important factor is the location of venules, which are the most affected vascular elements in inflamed tissue. There is a prevalence of large EAE susceptible venules traveling in the myelin rich white matter tracts in SJL/J mouse cerebellar cortex, indicating that the vascularization of this tissue may contribute to the increased susceptibility to inflammation in response to autoimmune attacks directed against CNS myelin.


The PD-1/PD-L1 complex resembles the antigen-binding Fv domains of antibodies and T cell receptors.
Signaling through the programmed death 1 (PD-1) inhibitory receptor upon binding its ligand, PD-L1, suppresses immune responses against autoantigens and tumors and plays an important role in the maintenance of peripheral immune tolerance. Release from PD-1 inhibitory signaling revives "exhausted" virus-specific T cells in chronic viral infections. Here we present the crystal structure of murine PD-1 in complex with human PD-L1. PD-1 and PD-L1 interact through the conserved front and side of their Ig variable (IgV) domains, as do the IgV domains of antibodies and T cell receptors. This places the loops at the ends of the IgV domains on the same side of the PD-1/PD-L1 complex, forming a surface that is similar to the antigen-binding surface of antibodies and T cell receptors. Mapping conserved residues allowed the identification of residues that are important in forming the PD-1/PD-L1 interface. Based on the structure, we show that some reported loss-of-binding mutations involve the PD-1/PD-L1 interaction but that others compromise protein folding. The PD-1/PD-L1 interaction described here may be blocked by antibodies or by designed small-molecule drugs to lower inhibitory signaling that results in a stronger immune response. The immune receptor-like loops offer a new surface for further study and potentially the design of molecules that would affect PD-1/PD-L1 complex formation and thereby modulate the immune response.


Five-Kilometer Time Trial Reliability of a Nonmotorized Treadmill and Comparison of Physiological and Perceptual Responses vs. a Motorized Treadmill.
Waldman, HS, Heatherly, AJ, Waddell, AF, Krings, BM, and O'Neal, EK. Five-kilometer time trial reliability of a nonmotorized treadmill and comparison of physiological and perceptual responses vs. a motorized treadmill. J Strength Cond Res 32(5): 1455-1461, 2018-This study examined the reliability of running performance across 3 nonmotorized treadmill (NMT) 5-km time trials (TTs) and physiological, gait, and perceptual differences at a 5-km pace for both NMT and motorized treadmills (MTs). Ten male runners experienced in road racing who had never run on an NMT completed 3 TT to establish personal best 5-km pace. In a later session, participants ran at this pace for 5 minutes on the NMT while metabolic, gait, and perceptual measures were recorded and then ran at outdoor 5-km personal best pace on an MT at 1% grade (counter-balanced crossover design). Intraclass correlation (ICC = 0.95) between the TT1 and TT2 was strong but improved between TT2 and TT3 (ICC = 0.99) with considerable reduction in variability. Nonmotorized treadmill resulted in a 24% slower pace (10.6 ± 1.5 vs. 13.9 ± 2.6 km·h; p < 0.001), shorter stride length (1.02 ± 0.10 vs. 1.27 ± 0.18 m; p < 0.001), and decreased cadence (175 ± 12 vs. 181 ± 13 steps per·minute; p = 0.01). However, V[Combining Dot Above]O2, respiratory exchange ratio (RER), lactate concentration, and heart rate did not differ between modalities (NMT = 3.4 ± 0.4 L·min, 0.96 ± 0.04, 6.9 ± 3.7 mmol, 172 ± 10 b·min; MT = 3.4 ± 0.5 L·min, 0.96 ± 0.04, 5.7 ± 3.4 mmol, 170 ± 10 b·min). rate of perceived exertion (RPE) for legs, breathing, and overall did not differ between treadmill types. A familiarization session should be included for TT using NMT. Other than gait and pace characteristics similar responses were elicited by both treadmills when running at 5-km pace. However, with these considerations, NMT TT of 4-km might be more appropriate in matching MT 5-km TT duration without altering physiological responses significantly.


Transgenic Bacillus thuringiensis corn hybrid performance against univoltine ecotype European corn borer (Lepidoptera: Crambidae) in South Dakota.
The performances of Bt-corn hybrids against univoltine ecotype European corn borer larvae were evaluated in South Dakota from 1997 to 1999. The corn hybrids were exposed to natural seasonal fluctuations of a univoltine ecotype European corn borer population. Larval injury parameters, grain yields, and gross incomes were quantified during the 3-yr study. In general, the Bt-corn hybrids had significantly higher yields than the untreated non-Bt isolines in 1997 and 1998 when corn borer pressures were high. However, most of the Bt-corn hybrids did not produce significant yield advantages in 1999 when the European corn borer pressure was low. Some of the Bt-corn hybrids even produced significantly lower yields than their untreated non-Bt counterparts in 1999. The performances of non-Bt isolines that were treated with permethrin granules at whorl stage were similar to Bt-corn hybrids in 1998 when the corn borer pressure was high and similar to the untreated non-Bt isolines in 1999 when the corn borer pressure was low. Injury-free corn produced by Bt-corn hybrids did not necessarily translate into higher yields in some hybrid groups. Grain moisture at harvest, which can result in moisture penalty or dockage, was significantly higher in most Bt-corn hybrids. Gross incomes of the Bt-corn hybrids were generally higher than the untreated non-Bt isolines when the corn borer infestation was high, but were either similar to or lower than the untreated non-Bt isolines when the corn borer infestation was low.


[3H]protein secretion in rat parotid gland: substance P-beta-adrenergic synergism.
In parotid fragment [3H]protein, secretion induced by substance P was moderate, but strongly Ca dependent. However, secretion induced by isoproterenol was large and Ca independent. Potentiation of protein secretion was observed when substance P (SP) and isoproterenol (ISO) acted together. Addition of 10(-8) M SP caused a shift to the left in the secretion dose-response curve caused by ISO, but did not enhance ISO-induced maximal response. The potentiating effect seems to be a postreceptor event, since it can be mimicked by forskolin (FK), known to induce directly cAMP accumulation, thus bypassing the beta-adrenergic receptor. The synergism described above was, therefore, investigated at the second messenger production level. Stimulation of parotid gland fragments by simultaneous addition of SP plus ISO or FK did not modify cAMP nor inositol trisphosphate (IP3) accumulation induced independently by each secretagogue alone. The ionophore A23187 was also able to potentiate secretion induced by a beta-adrenergic agonist, this effect being totally abolished by external calcium omission, thus suggesting a role for external calcium in this potentiation phenomenon. These results suggest that the potentiation phenomenon observed is a postreceptor event that occurs at a step distal from the second messenger production.


FK-506 delays corneal graft rejection in a model of corneal xenotransplantation.
FK-506 is a relatively new immunosuppressant similar in action to cyclosporine A, but is much more potent. Its primary action is against T lymphocytes, the major cellular component in corneal allograft rejection. The purpose of this study was the evaluation of the ability of topical and systemic FK-506 in preventing corneal xenograft rejection in an experimental animal model. Cross-species xenotransplants were used as the most vigorous stimulus to induce corneal rejection. Corneas derived from Hartley guinea pigs were transplanted into the left eyes of 32 male Lewis rats. Topical treatment was administered by using FK-506 0.3 mg/ml in a cyclodextrin suspension or vehicle (cyclodextrin suspension) four times per day. For systemic treatment, 0.5 mg/kg/day of FK-506 or vehicle (saline) was administered intraperitoneally. Treatments were started 60 minutes after surgery and continued for 21 days. The grafts underwent a double-masked examination, and a score was given for clarity, edema, and vascularization. The animals were sacrificed 21 days after transplantation. The control groups had allograft rejection after 6.75 +/- 0.31 (topical vehicle) and after 7.37 +/- 0.32 (systemic vehicle) days. The FK-506-treated groups showed allograft rejection after 14 +/- 0.88 (topical FK-506) or after 16.25 +/- 1.23 (systemic FK-506) days. In addition, FK-506-treated rats manifested less corneal neovascularization than control animals. We conclude that systemic or topical FK-506 is effective in prolonging xenograft survival in the rat keratoplasty model.


Acute high-fat feeding does not prevent the improvement in glucose tolerance after resistance exercise in lean individuals.
Our first aim was to investigate whether the ingestion of a single high-fat meal impairs glucose tolerance. Our second aim was to investigate whether improvements in glucose tolerance that are seen after resistance exercise remain when exercise is performed after ingestion of a high-fat meal. Eight young males consumed either a high fat (HF) or an isocaloric control (CON) meal in the morning and underwent an oral glucose tolerance test (OGTT) 6 h later. On two other occasions, a single 1 h bout of resistance exercise was completed 2 h after consumption of each meal (HFE and CONE). There were no significant differences in plasma glucose and plasma insulin areas under the curve (AUC) or estimates of insulin sensitivity between the HF and CON trials (P > 0.05). The HFE and CONE trials elicited a ~20% lower plasma glucose AUC (P < 0.05) compared to their respective control trials. The HFE also elicited a ~25% lower plasma insulin AUC (P < 0.05) in comparison to the HF trial. The HFE trial also significantly improved estimates of insulin sensitivity in comparison to the HF condition (P < 0.05). In conclusion, this study demonstrates that consumption of a single HF meal does not impair glucose tolerance in the resting state in lean individuals and that an acute bout of resistance exercise remains effective in enhancing glucose tolerance following the ingestion of a single high-fat meal.


[Improvement of public health in London in the nineteenth century and the probably limited role of the new sewage system].
--London was one of the most rapidly expanding cities in the world in the nineteenth century, but the water supply and sanitary conditions were extremely poor. --In the nineteenth century, there were many theories regarding the causes and spread of epidemic diseases. The miasma theory, which postulated that disease was the result of bad air and odours, was the most popular. --In 1858, as a result of the foul smells from the Thames, the decision was made to improve the sewage system. The new system was first used in 1865, but its actual effect on public health is not clear. --The life expectancy of Londoners did not change in the period 1826-1871, but increased substantially thereafter. In view of this time window, this may have been a result of improvements in the sewage system. --However, it is likely that several other factors, such as improvements in the food and water supply, vaccination programmes, and natural circumstances, were responsible for a large part of this increase.