Combinatorial QSAR modeling of chemical toxicants tested against Tetrahymena pyriformis.
Selecting most rigorous quantitative structure-activity relationship (QSAR) approaches is of great importance in the development of robust and predictive models of chemical toxicity. To address this issue in a systematic way, we have formed an international virtual collaboratory consisting of six independent groups with shared interests in computational chemical toxicology. We have compiled an aqueous toxicity data set containing 983 unique compounds tested in the same laboratory over a decade against Tetrahymena pyriformis. A modeling set including 644 compounds was selected randomly from the original set and distributed to all groups that used their own QSAR tools for model development. The remaining 339 compounds in the original set (external set I) as well as 110 additional compounds (external set II) published recently by the same laboratory (after this computational study was already in progress) were used as two independent validation sets to assess the external predictive power of individual models. In total, our virtual collaboratory has developed 15 different types of QSAR models of aquatic toxicity for the training set. The internal prediction accuracy for the modeling set ranged from 0.76 to 0.93 as measured by the leave-one-out cross-validation correlation coefficient ( Q abs2). The prediction accuracy for the external validation sets I and II ranged from 0.71 to 0.85 (linear regression coefficient R absI2) and from 0.38 to 0.83 (linear regression coefficient R absII2), respectively. The use of an applicability domain threshold implemented in most models generally improved the external prediction accuracy but at the same time led to a decrease in chemical space coverage. Finally, several consensus models were developed by averaging the predicted aquatic toxicity for every compound using all 15 models, with or without taking into account their respective applicability domains. We find that consensus models afford higher prediction accuracy for the external validation data sets with the highest space coverage as compared to individual constituent models. Our studies prove the power of a collaborative and consensual approach to QSAR model development. The best validated models of aquatic toxicity developed by our collaboratory (both individual and consensus) can be used as reliable computational predictors of aquatic toxicity and are available from any of the participating laboratories.


Induced hydraulic pumping via integrated submicrometer cylindrical glass capillaries.
Here, we report on a micropump that generates hydraulic pressure owing to a mismatch in EOF rates of microchannels and submicrometer cylindrical glass capillaries integrated on silicon. The electrical conductance of such capillaries in the dilute limit departs from bulk linear behavior as well as from the surface-charge-governed saturation in nanoslits that is well described by the assumption of a constant surface charge density. The capillaries show rather a gradual decrease in conduction at low salt concentrations, which can be explained more aptly by a variable surface charge density that accounts for chemical equilibrium of the surface. The micropump uses a traditional cross-junction structure with ten identical capillaries integrated in parallel on a side arm and each with a 750 nm diameter and 3 mm length. For an applied voltage of 700 V, a hydraulic pressure up to 5 kPa is generated with a corresponding flow velocity nearly 3 mm/s in a straight field-free branch 20 μm wide, 10 μm deep, and 10 mm long. The micropump utility has been demonstrated in an open tubular LC of three fluorescently labeled amino acids in just less than 20 s with minimal plate height values between 3 and 7 μm. The submicrometer capillaries are self-enclosed and produced through a unique process that does not require high-resolution advanced lithography or wafer-bonding techniques to define their highly controlled precise structures.


Focal adhesion signaling: vascular smooth muscle cell contractility beyond calcium mechanisms.
Smooth muscle cell (SMC) contractility is essential to vessel tone maintenance and blood pressure regulation. In response to vasoconstrictors, calcium-dependent mechanisms promote the activation of the regulatory myosin light chain, leading to increased cytoskeleton tension that favors cell shortening. In contrast, SMC maintain an intrinsic level of a contractile force independent of vasoconstrictor stimulation and sustained SMC contraction beyond the timescale of calcium-dependent mechanisms suggesting the involvement of additional players in the contractile response. Focal adhesions (FAs) are conceivable candidates that may influence SMC contraction. They are required for actin-based traction employed by cells to sense and respond to environmental cues in a process termed mechanotransduction. Depletion of FA proteins impairs SMC contractility, producing arteries that are prone to dissection because of a lack of mechanical stability. Here, we discuss the role of calcium-independent FA signaling mechanisms in SMC contractility. We speculate that FA signaling contributes to the genesis of a variety of SMC phenotypes and discuss the potential implications for mechanical homeostasis in normal and diseased states.


Landscape-scale conservation design across biotic realms: sequential integration of aquatic and terrestrial landscapes.
Systematic conservation planning has been used extensively throughout the world to identify important areas for maintaining biodiversity and functional ecosystems, and is well suited to address large-scale biodiversity conservation challenges of the twenty-first century. Systematic planning is necessary to bridge implementation, scale, and data gaps in a collaborative effort that recognizes competing land uses. Here, we developed a conservation planning process to identify and unify conservation priorities around the central and southern Appalachian Mountains as part of the Appalachian Landscape Conservation Cooperative (App LCC). Through a participatory framework and sequential, cross-realm integration in spatial optimization modeling we highlight lands and waters that together achieve joint conservation goals from LCC partners for the least cost. This process was driven by a synthesis of 26 multi-scaled conservation targets and optimized for simultaneous representation inside the program Marxan to account for roughly 25% of the LCC geography. We identify five conservation design elements covering critical ecological processes and patterns including interconnected regions as well as the broad landscapes between them. Elements were then subjected to a cumulative threats index for possible prioritization. The evaluation of these elements supports multi-scaled decision making within the LCC planning community through a participatory, dynamic, and iterative process.


Simultaneous analysis by LC-MS/MS of 22 ketosteroids with hydroxylamine derivatization and underivatized estradiol from human plasma, serum and prostate tissue.
This study describes a validated LC-MS/MS method for assaying 23 steroids within a single run from 150 μl of human plasma, serum or prostatic tissue homogenate. Isotope-labeled steroids were used as internal standards. Samples were extracted with toluene, and ketosteroids were derivatized with hydroxylamine prior to LC-MS/MS analysis. The steroids were separated on a C18 column and methanol was used as an organic solvent with the addition of 0.2 mM ammonium fluoride to improve underivatized estradiol (E2) ionization. Certified reference serums as well as plasma samples, and homogenates of prostate tissue were utilized in the method validation. The specificity of the method was inspected with a total of 27 steroids. The validation proved that the method was suitable for the quantitative analysis of a wide panel of androgens (testosterone, T (3.3 pM-13 nM); androstenedione, A4 (3.3 pM-13 nM); 5α-androstanedione, DHA4 (13 pM-13 nM); dehydroepiandrosterone, DHEA (67 pM-133 nM); dihydrotestosterone, DHT (33 pM-33 nM); 11-ketodihydrotestosterone, 11KDHT (13 pM-13nM); 11-ketotestosterone, 11KT (33 pM-6.7 nM); 11β-hydroxyandrostenedione, 11bOHA4 (33 pM-13 nM); 11β-hydroxytestosterone, 11OHT (13 pM-33 nM)), as well as estrogens (estrone, E1 (3.3 pM-13 nM)), progestagens (17α-hydroxypregnenolone, 17OHP5 (32 pM-127 nM); 17α-hydroxyprogesterone, 17OHP4 (67 pM-133 nM); progesterone, P4 (3.3 pM-13 nM); pregnenolone, P5 (6.6 pM-13 nM)), and glucocorticoids (cortisol, F (33 pM-134 nM); cortisone E (66 pM-131 nM); corticosterone, B (33 pM-67 nM); 11-deoxycortisol, S (33 pM-66 nM); 21-hydroxyprogesterone, 21OHP4 (32 pM-13 nM)). Furthermore, E2 (335 pM-134 nM) and 11α-hydroxyandrostenedione, 11aOHA4 (33 pM-33 nM) could be analyzed if the concentration in the sample was high enough. In addition, aldosterone, A (128 pM-64 nM) and 11-ketoandrostenedione, 11KA4 (33 pM-13 nM) could be analyzed semiquantitatively. The limits of quantification for all compounds ranged from 0.9 to 91 pg/ml, and from 0.009 to 0.9 pg/mg tissue. Compared to our previous method, this new method also permits the analysis of the more challenging steroids, like DHT, DHEA and P5, and a panel of 11-ketosteroids.


Complementary and alternative medicine practice and perceptions of attendees of primary care centers in Eastern Saudi Arabia.
Complementary and alternative medicine (CAM) refers to the use of nonconventional medical practices together with standard medical care. Specific forms of CAM included in this study are the use of honey, dry and wet Hijama, Quran, Zamzam water, Nigella sativa (black seed), and others. The objective was to determine the prevalence of the use of different CAM modalities by the attendees of primary health-care centers (PHCCs) and assess the reasons behind their use. A cross-sectional study was conducted at PHCCs in the cities of Dammam and Al Khobar, over a period of 12 months. The study population was patients and their accompanying relatives attending the PHCCs, aged 18 years and above. Data were collected using a self-administered questionnaire in Arabic designed by the investigators after a review of the literature on the use of CAM. Questionnaire was modified after pilot testing among 200 medical students, and was validated by two expert consultants of the family and community medicine department at the university. The questionnaire was administered and supervised by the 3rd and 4th year medical students. Data were entered and analyzed using the SPSS version 16 (SPSS Inc., Chicago, IL, USA) statistical software. Descriptive statistics were computed, and odds ratios were calculated for associations between CAM use and various independent factors. Logistic regression analysis was used to determine the factors which predict the use of CAM by attendees. All the analyses were performed at α =0.05. The total number of attendees interviewed was 2114; 52.5% of whom reported using CAM. The types of CAM varied in their popularity of use, the majority of users favoring honey and Quran, 24.6% and 20.5%, respectively. Recommendations by relatives and friends were reported as the most common reason in this sample for using CAM (56.1%). Age, awareness of CAM, perceptions that use of CAM is better, use of CAM helps, herbs had no side effects, CAM was easy to obtain, and the attendee currently suffers from a disease were significantly associated with higher CAM use. More than 50% of the participants reported using CAM; honey and Quran were the most widely used modalities. For people to make informed decisions on the use of complementary medicine, physicians should impress on their patients to inform their doctors of their use of complementary medicine modalities so that encouragement could be given where appropriate.


Reduced Corneal Endothelial Cell Density in Patients With Dry Eye Disease.
To evaluate corneal endothelial cell density (ECD) in patients with dry eye disease (DED) compared to an age-matched control group. Cross-sectional, controlled study. This study included 90 eyes of 45 patients with moderate to severe DED (aged 53.7 ± 9.8 years) and 30 eyes of 15 normal controls (aged 50.7 ± 9.8 years). All subjects had a complete ophthalmic evaluation including symptom assessment using the Ocular Surface Disease Index (OSDI) and corneal fluorescein staining. In addition, laser scanning in vivo confocal microscopy was performed to measure the density of the following parameters in the central cornea: endothelial cells, subbasal nerves, and subbasal immune dendritic cells. Corneal ECD was significantly lower in the DED group (2595.8 ± 356.1 cells/mm(2)) than in the control group (2812.7 ± 395.2 cells/mm(2), P = .046). The DED group showed significantly lower corneal subbasal nerve density (17.1 ± 6.9 mm/mm(2)) compared to the control group (24.7 ± 4.4 mm/mm(2), P < .001). Dendritic cell density was significantly higher in the DED group than in the controls (111.7 ± 137.3 vs 32.0 ± 24.4 cells/mm(2), respectively, P = .002). There were statistically significant correlations between corneal ECD and dry eye severity parameters including the OSDI score (rs = -0.26, P = .03), and corneal fluorescein staining (rs = -0.28, P = .008). There is a significant reduction in corneal ECD in DED that correlates with clinical severity of the disease.


Relation between Respiratory Sinus Arrythymia and Startle Response during Predictable and Unpredictable Threat.
Research suggests that lower respiratory sinus arrhythmia (RSA) is associated with greater aversive responding. One physiological indicator of aversive responding is startle potentiation. While a few studies have demonstrated an inverse association between RSA and startle potentiation, no study to date has distinguished whether this relation is similar for predictable versus unpredictable aversive stimuli. This is an important distinction, given that degree of predictability has been shown to be an important determinant of aversive responding. The present study examined whether resting RSA was associated with startle eye blink responding during predictable and unpredictable threat of electric shock. Resting RSA was collected during a 6-minute seated baseline phase at the beginning of the experimental session. Participants then completed a computerized startle task in which predictable and unpredictable shocks were administered. Results indicated that lower resting RSA was associated with greater startle potentiation during unpredictable threat, but not during predictable threat. These findings are consistent with a growing body of literature suggesting that individual differences in RSA are associated with aversive responding, and extend previous work by suggesting that RSA may be more robustly associated with a heightened sensitivity to unpredictable threat. This pattern of results may have implications for the understanding of pathological anxiety given that individuals with anxiety disorders typically exhibit low RSA and heightened responding during unpredictable threat.


Recovery of Lactobacillus rhamnosus GG from human colonic biopsies.
The colonization of Lactobacillus rhamnosus GG (ATCC 53103, henceforth L.GG) in five human colonoscopy patients was studied. The test subjects consumed whey drink fermented with the bacterium for 12 d before the colonoscopy. The presence of L.GG was subsequently checked both in the faecal samples and in the colonic biopsies obtained from various locations in the large intestine. In all patients L.GG was the dominant faecal lactic acid bacterium as a result of the administration. In four patients L.GG could also be recovered from the biopsies, while with one patient (suffering from ulcerative colitis diagnosed during the colonoscopy) no L.GG was detected in the biopsy samples. The results suggest that L.GG is able to adhere in vivo to the colon. Study of the faecal samples alone is apparently not sufficient for elucidation of the gastrointestinal ecology of probiotic bacteria.


Bacterial coinfection in critically ill COVID-19 patients with severe pneumonia.
Severe 2019 novel coronavirus infectious disease (COVID-19) with pneumonia is associated with high rates of admission to the intensive care unit (ICU). Bacterial coinfection has been reported to be rare. We aimed at describing the rate of bacterial coinfection in critically ill adult patients with severe COVID-19 pneumonia. All the patients with laboratory-confirmed severe COVID-19 pneumonia admitted to the ICU of Tenon University-teaching hospital, from February 22 to May 7th, 2020 were included. Respiratory tract specimens were obtained within the first 48 h of ICU admission. During the study period, 101 patients were referred to the ICU for COVID-19 with severe pneumonia. Most patients (n = 83; 82.2%) were intubated and mechanically ventilated on ICU admission. Overall, 20 (19.8%) respiratory tract specimens obtained within the first 48 h. Staphylococcus aureus was the main pathogen identified, accounting for almost half of the early-onset bacterial etiologies. We found a high prevalence of early-onset bacterial coinfection during severe COVID-19 pneumonia, with a high proportion of S. aureus. Our data support the current WHO guidelines for the management of severe COVID-19 patients, in whom antibiotic therapy directed to respiratory pathogens is recommended.


Development and initial validation of a project-based rubric to assess the systems-based practice competency of residents in the clinical chemistry rotation of a pathology residency.
Systems-based practice (SBP) is 1 of 6 core competencies required in all resident training programs accredited by the Accreditation Council for Graduate Medical Education. Reliable methods of assessing resident competency in SBP have not been described in the medical literature. To develop and validate an analytic grading rubric to assess pathology residents' analyses of SBP problems in clinical chemistry. Residents were assigned an SBP project based upon unmet clinical needs in the clinical chemistry laboratories. Using an iterative method, we created an analytic grading rubric based on critical thinking principles. Four faculty raters used the SBP project evaluation rubric to independently grade 11 residents' projects during their clinical chemistry rotations. Interrater reliability and Cronbach α were calculated to determine the reliability and validity of the rubric. Project mean scores and range were also assessed to determine whether the rubric differentiated resident critical thinking skills related to the SBP projects. Overall project scores ranged from 6.56 to 16.50 out of a possible 20 points. Cronbach α ranged from 0.91 to 0.96, indicating that the 4 rubric categories were internally consistent without significant overlap. Intraclass correlation coefficients ranged from 0.63 to 0.81, indicating moderate to strong interrater reliability. We report development and statistical analysis of a novel SBP project evaluation rubric. The results indicate the rubric can be used to reliably assess pathology residents' critical thinking skills in SBP.


Effect of age on calcium and phosphate absorption. Role of 1,25-dihydroxyvitamin D.
In humans, there is a decrease in intestinal absorption of Ca with age and a decrease in the capacity to adapt to a low Ca diet. The rat has been used as an animal model in which to study the mechanisms responsible for this decreased absorption and adaptation with age. Intestinal absorption of Ca and Pi declines with age in the rat. The decline is primarily in the energy- and vitamin D-dependent component of transport found in the duodenum and, in the case of Pi transport, in the proximal jejunum. Administration of 1,25-dihydroxyvitamin D (1,25(OH)2D) increases Ca and P transport in all age groups, but the maximal levels attained are less in older animals compared to the young. Feeding a low-Ca diet increases serum 1,25(OH)2D levels and intestinal Ca absorption only in young animals. However, a low-Ca diet increases intestinal Pi absorption in all age groups, even in the absence of elevated serum 1,25(OH)2D levels. Knowledge of the mechanisms responsible for the age-related decrease in Ca and Pi absorption may suggest new ways of improving mineral absorption in the elderly.


Sustainable antimicrobial effect of silver sulfadiazine-loaded nanosheets on infection in a mouse model of partial-thickness burn injury.
Partial-thickness burn injury has the potential for reepithelialization and heals within 3weeks. If the wound is infected by bacteria before reepithelization, however, the depth of disruption increases and the lesion easily progresses to the full-thickness dermal layers. In the treatment of partial-thickness burn injury, it is important to prevent the wound area from bacterial infection with an antimicrobial dressing. Here, we have tested the antimicrobial properties of polymeric ultra-thin films composed of poly(lactic acid) (termed "PLA nanosheets"), which have high flexibility, adhesive strength and transparency, and silver sulfadiazine (AgSD), which exhibits antimicrobial efficacy. The AgSD-loaded nanosheet released Ag(+) for more than 3days, and exerted antimicrobial efficacy against methicillin-resistant Staphylococcus aureus (MRSA) in an in vitro Kirby-Bauer test. By contrast, a cell viability assay indicated that the dose of AgSD used in the PLA nanosheets did not show significant cytotoxicity toward fibroblasts. In vivo evaluation using a mouse model of infection in a partial-thickness burn wound demonstrated that the nanosheet significantly reduced the number of MRSA bacteria on the lesion (more than 10(5)-fold) and suppressed the inflammatory reaction, thereby preventing a protracted wound healing process.


Sub-classification of Advanced-Stage Hepatocellular Carcinoma: A Cohort Study Including 612 Patients Treated with Sorafenib.
Advanced hepatocellular carcinoma (HCC) is associated with various clinical conditions including major vessel invasion, metastasis, and poor performance status. The aim of this study was to establish a prognostic scoring system and to propose a sub-classification of the Barcelona-Clinic Liver Cancer (BCLC) stage C. This retrospective study included consecutive patientswho received sorafenib for BCLC stage C HCC at a single tertiary hospital in Korea. A Cox proportional hazard model was used to develop a scoring system, and internal validationwas performed by a 5-fold cross-validation. The performance of the model in predicting risk was assessed by the area under the curve and the Hosmer-Lemeshow test. A total of 612 BCLC stage C HCC patients were sub- classified into strata depending on their performance status. Five independent prognostic factors (Child-Pugh score, α-fetoprotein, tumor type, extrahepatic metastasis, and portal vein invasion) were identified and used in the prognostic scoring system. This scoring system showed good discrimination (area under the receiver operating characteristic curve, 0.734 to 0.818) and calibration functions (both p < 0.05 by the Hosmer-Lemeshow test at 1 month and 12 months, respectively). The differences in survival among the different risk groups classified by the total score were significant (p < 0.001 by the log-rank test in both the Eastern Cooperative Oncology Group 0 and 1 strata). The heterogeneity of patientswith BCLC stage C HCC requires sub-classification of advanced HCC. A prognostic scoring system with five independent factors is useful in predicting the survival of patients with BCLC stage C HCC.


The topology of chromatin-binding domains in the NuRD deacetylase complex.
Class I histone deacetylase complexes play essential roles in many nuclear processes. Whilst they contain a common catalytic subunit, they have diverse modes of action determined by associated factors in the distinct complexes. The deacetylase module from the NuRD complex contains three protein domains that control the recruitment of chromatin to the deacetylase enzyme, HDAC1/2. Using biochemical approaches and cryo-electron microscopy, we have determined how three chromatin-binding domains (MTA1-BAH, MBD2/3 and RBBP4/7) are assembled in relation to the core complex so as to facilitate interaction of the complex with the genome. We observe a striking arrangement of the BAH domains suggesting a potential mechanism for binding to di-nucleosomes. We also find that the WD40 domains from RBBP4 are linked to the core with surprising flexibility that is likely important for chromatin engagement. A single MBD2 protein binds asymmetrically to the dimerisation interface of the complex. This symmetry mismatch explains the stoichiometry of the complex. Finally, our structures suggest how the holo-NuRD might assemble on a di-nucleosome substrate.


High temperature radiofrequency transurethral ablation of the prostate (TURAPY).
Conventional treatment for BPH in recent decades has been transurethral resection of the prostate (TURP) or open prostatectomy, depending on adenoma volume. As TURP is a surgical treatment associated with high morbidity, a certain failure rate and mortality, urologists have been looking for other alternative treatments that are safer and at least as effective. Prostatic tissue ablation is one of the preferred alternative BPH treatments today and can be achieved with laser or with TURAPY--Transurethral Ablation Prostatectomy. This new treatment based on thermeablation of the prostate has been introduced recently by Direx Medical Systems. The treatment is administered with a transurethral Folley-like electrode delivering temperatures in the range of 80 degrees C. TURAPY causes extensive ablation of the tissue around the prostatic urethra, without the complications of a surgical procedure. Twenty two men, age 76 to 92 years (mean 79) with symptomatic Benign Prostatic Hyperplasia (BPH) and suprapubuc urinary fistulas were treated with the computer controlled radio frequency device TURAPY as day cases under local anaesthesia. In 1, 5-2 months after the procedure urination was restored, urinary fistulas healed.


Sporting activity and drug use: Alcohol, cigarette and cannabis use among elite student athletes.
To study the relationship between sporting activity and alcohol, cigarette and cannabis use among adolescents and young adults, by focusing on elite student athletes (ESAs). Cross-sectional survey (Spring 2002), in a sample of 460 ESAs (ages 16-24 years) recruited at 40 public centres gathering the young sporting elite from 30 different sports in South-Eastern France, comparison with samples of the general population of adolescents in South-Eastern France. Respondents were asked confidentially by a self-administered questionnaire about their use of licit and illicit drugs, their sporting activity and other aspects of their life-style. Prevalences of cigarette, alcohol and cannabis use were markedly lower for ESAs than for other adolescents (generally twice or three times as low). Among ESAs, when compared with the practice of an individual sport, the practice of a team sport was correlated positively with alcohol use (OR = 2.7 for girls, OR = 1.8 for boys), and the practice of a sliding sport was correlated positively with cannabis use (for girls: OR = 2.3) and with alcohol use (for boys: 4.3). Girls who entered competition at international level were more prone to smoke cigarettes and cannabis (OR = 6.1 and 2.4, respectively). As a whole, practising sports as an elite student-athlete can be considered as correlated negatively with cigarette, alcohol and cannabis use. Nevertheless, this relationship depends on the kind of sport practised as well as the level of competition, and further research is needed to understand specific elite athletes' motives for use.


Good-quality diet in the early years may have a positive effect on academic achievement.
The aim of this study was to investigate the associations between early diet and academic performance during childhood. Participants were from the Western Australian Pregnancy Cohort (Raine) Study (n = 2287). Frequency of consumption of food and beverages was collected at the one-, two- and three-year follow-ups, using a 24-hour food recall. Diet scores were developed from the number of eating occasions. The Western Australian Literacy and Numeracy Assessment (WALNA) data from grades five (age 10) and seven (age 12) were linked to the Raine study using The Western Australian Data Linkage System. The association between diet scores and WALNA scores was assessed using multivariate linear regression models. A higher (i.e. better quality) diet score at one year of age was associated with significantly higher scores in mathematics, reading, writing and spelling at both grades five and seven. Associations were observed between a higher diet score at two years and academic scores for mathematics, writing and spelling at grade seven. Higher dairy consumption at ages one, two and three, and higher fruit consumption at age one were associated with higher academic scores at all ages. Quality of early diet may be a predictor for later academic achievement.


School-based multidisciplinary student-led clinics in health and Australian accreditation standards: A scoping review.
Student-led clinics can provide health services to marginalised groups where service offerings are sparse or difficult to access, such as rural areas. Offering these services to children and young people can promote health and well-being by addressing the individual challenges and the social determinants of health. There is uncertainty, however, as to whether student-led clinics can meet Australian accreditation standards for health professionals completing degree programs. This study aims to determine the capacity for health student placements in school-based student-led clinics to meet accreditation standards. A systematic scoping review was conducted based on Arksey and O'Malley's framework and the PRISMA-ScR statement. Several databases were examined, including Ebsco (Academic Source and CINAHL), ProQuest (PsycINFO, ERIC) and grey literature sources along with a desktop review of accreditation standards across seven health disciplines. Two independent reviewers screened eligible studies. The search retrieved 1037 records with 65 full-text papers assessed for eligibility. Eleven papers met the inclusion criteria. Based on the evidence, both nursing and exercise and sports science accreditation standards were best suited to student-led clinics. Although broad categories of work-integrated learning activities were applied, it appears feasible to expect accreditation standards for health disciplines at an Australian university to be a good fit for health student-led school-based clinics. Increasing health student placement opportunities within student-led clinics can improve the health and well-being of children and young people in regional, rural and remote (RRR) areas of Australia who may otherwise have limited access to allied health services.


Ergonomics and Related Physical Symptoms Among Neurosurgeons.
Work-related musculoskeletal disorders (WMSDs) among neurosurgeons can lead to consequences for themselves, the hospital, and society. In the current study, the working conditions of neurosurgeons from an ergonomic point of view is evaluated, together with WMSD. Members of the Congress of Neurological Surgeons were surveyed with an online survey covering 1) demographics, 2) working conditions, 3) ergonomic features during 3 neurosurgical procedures, and 4) whether they experienced WMSD. Multivariate logistic regression analysis was performed to identify factors associated with experiencing WMSD. In total, 417 replies were received. Respondents had a mean tenure of 21.9 years. In total, 33.1% stated that the operating room is furnished ergonomically and 90.7% stated that ergonomics is an underexposed field in neurosurgery. The majority of the respondents (73.6%) had experienced WMSD. Performing long procedures and spine surgery were mentioned most often as cause for WMSD. Due to WMSD, 11.3% of the respondents had to take time off work, and 14.2% considered changing their career. Analgesics were mostly used as treatment for WMSD (42.9%) and 7.4% underwent surgery. Having a tenure ≤15 years and having the operating room furnished ergonomically were associated with less WMSD in univariate analysis, whereas only a tenure <15 years was in multivariate analysis (odds ratio 0.27; 95% confidence interval 0.085-0.831). Despite only a minority of the surgeons taking time off due to WMSD, the majority suffers from WMSD. Education of residents in ergonomics to prevent WMSD in their later careers and ergonomic furnishing of surgical instrumentation and operating rooms seem to be areas for improvement.


Donor B-cell alloantibody deposition and germinal center formation are required for the development of murine chronic GVHD and bronchiolitis obliterans.
Chronic GVHD (cGVHD) poses a significant risk for HSCT patients. Preclinical development of new therapeutic modalities has been hindered by models with pathologic findings that may not simulate the development of human cGVHD. Previously, we have demonstrated that cGVHD induced by allogeneic HSCT after a conditioning regimen of cyclophosphamide and total-body radiation results in pulmonary dysfunction and airway obliteration, which leads to bronchiolitis obliterans (BO), which is pathognomonic for cGVHD of the lung. We now report cGVHD manifestations in a wide spectrum of target organs, including those with mucosal surfaces. Fibrosis was demonstrated in the lung and liver and was associated with CD4(+) T cells and B220(+) B-cell infiltration and alloantibody deposition. Donor bone marrow obtained from mice incapable of secreting IgG alloantibody resulted in less BO and cGVHD. Robust germinal center reactions were present at the time of cGVHD disease initiation. Blockade of germinal center formation with a lymphotoxin-receptor-immunoglobulin fusion protein suppressed cGVHD and BO. We conclude that cGVHD is caused in part by alloantibody secretion, which is associated with fibrosis and cGVHD manifestations including BO, and that treatment with a lymphotoxin-β receptor-immunoglobulin fusion protein could be beneficial for cGVHD prevention and therapy.


CircHDAC9 regulates myocardial ischemia-reperfusion injury via miR-671-5p/SOX4 signaling axis.
Myocardial ischemia-reperfusion (I/R), a harmful process in the treatment of cardiovascular diseases, can cause secondary damage to the cardiac tissues. Circular RNAs (circRNAs) are important regulators in a number of cardiac disorders. However, the role of circHDAC9 in myocardial I/R injury has not been clarified. Human cardiac myocytes (HCMs) were treated with hypoxia/reoxygenation (H/R) and mice were subjected to I/R. Quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to analyze the expression of circHDAC9, miR-671-5p, and SOX4, and western blot was used to detect SOX4 protein. The binding relationship among circHDAC9, miR-671-5p, and SOX4 was confirmed by RNA pull-down, luciferase, and RNA immunoprecipitation (RIP) assays. The effects of circHDAC9/miR-671-5p/SOX4 axis on the apoptosis, oxidative stress and inflammation were evaluated in both myocardial I/R injury models. The expression of circHDAC9 and SOX4 was noticeably elevated, whereas miR-671-5p expression was downregulated in both myocardial I/R injury models. circHDAC9 knockdown significantly reduced the apoptosis, activities of caspase-3 and caspase-9, ROS intensity, MDA activity, and concentrations of TNF-α, IL-1β, and IL-6, but increased the viability and SOD activity in H/R-treated HCMs. Suppression of circHDAC9 dramatically reduced the levels of circHDAC9 and SOX4, while enhanced miR-671-5p expression in H/R-treated HCMs. CircHDAC9 functioned via sponging miR-671-5p to regulate SOX4 expression in vitro. Additionally, silencing of circHDAC9 improved the pathological abnormalities and cardiac dysfunction, and reduced the apoptosis, oxidative stress and inflammation in mice with myocardial I/R injury. Inhibition of circHDAC9 significantly improved myocardial I/R injury by regulating miR-671-5p/SOX4 signaling pathway.


APC and EB1 function together in mitosis to regulate spindle dynamics and chromosome alignment.
Recently, we have shown that a cancer causing truncation in adenomatous polyposis coli (APC) (APC(1-1450)) dominantly interferes with mitotic spindle function, suggesting APC regulates microtubule dynamics during mitosis. Here, we examine the possibility that APC mutants interfere with the function of EB1, a plus-end microtubule-binding protein that interacts with APC and is required for normal microtubule dynamics. We show that siRNA-mediated inhibition of APC, EB1, or APC and EB1 together give rise to similar defects in mitotic spindles and chromosome alignment without arresting cells in mitosis; in contrast inhibition of CLIP170 or LIS1 cause distinct spindle defects and mitotic arrest. We show that APC(1-1450) acts as a dominant negative by forming a hetero-oligomer with the full-length APC and preventing it from interacting with EB1, which is consistent with a functional relationship between APC and EB1. Live-imaging of mitotic cells expressing EB1-GFP demonstrates that APC(1-1450) compromises the dynamics of EB1-comets, increasing the frequency of EB1-GFP pausing. Together these data provide novel insight into how APC may regulate mitotic spindle function and how errors in chromosome segregation are tolerated in tumor cells.


A Pilot Study of Horizontal Head and Eye Rotations in Baseball Batting.
The purpose of the study was to measure and compare horizontal head and eye tracking movements as baseball batters "took" pitches and swung at baseball pitches. Two former college baseball players were tested in two conditions. A pitching machine was used to project tennis balls toward the subjects. In the first condition, subjects acted as if they were taking (i.e., not swinging) the pitches. In the second condition, subjects attempted to bat the pitched balls. Head movements were measured with an inertial sensor; eye movements were measured with a video eye tracker. For each condition, the relationship between the horizontal head and eye rotations was similar for the two subjects, as were the overall head-, eye-, and gaze-tracking strategies. In the "take" condition, head movements in the direction of the ball were larger than eye movements for much of the pitch trajectory. Large eye movements occurred only late in the pitch trajectory. Gaze was directed near the ball until approximately 150 milliseconds before the ball arrived at the batter, at which time gaze was directed ahead of the ball to a location near that occupied when the ball crosses the plate. In the "swing" condition, head movements in the direction of the ball were larger than eye movements throughout the pitch trajectory. Gaze was directed near the ball until approximately 50 to 60 milliseconds prior to pitch arrival at the batter. Horizontal head rotations were larger than horizontal eye rotations in both the "take" and "swing" conditions. Gaze was directed ahead of the ball late in the pitch trajectory in the "take" condition, whereas gaze was directed near the ball throughout much of the pitch trajectory in the "swing" condition.


[A case of fatal disseminated infection caused by Mycobacterium bovis BCG strain and the identification of the isolate by spoligotyping].
The vaccine strain Mycobacterium bovis BCG may lead to disseminated infection in patients with immune deficiency. In this report a patient who developed fatal disseminated tuberculosis caused by M. bovis BCG strain was presented. One year old male patient with the previous history of recurrent lower respiratory tract infection, was admitted to the hospital with the complaints of fever, cough and diarrhea continuing for 20 days. There was no family history of tuberculosis or history of contact with a tuberculosis case. Physical examination of the case revealed growth retardation and reticular and reticulonodular infiltration was detected in his chest X-ray. The results of sweat test, cystic fibrosis gene mutation analysis and metabolic screening tests were normal. Since fever continued and infiltrations persisted in the chest X-ray despite antibiotic therapy, PPD test was applied and acid-fast bacilli (AFB) were investigated in his gastric aspirate and stool samples for three consecutive days. PPD test was negative and no AFB were detected in the microscopic examination of the clinical samples. However, growth in Lowenstein-Jensen medium was detected in the stool sample on the 38th day of incubation. The antimycobacterial susceptibility testing performed at BACTEC MGIT (Mycobacterial Growth Indicator Tube) 960 system (Becton-Dickinson, USA) revealed that the isolate was susceptible to rifampin, isoniazid, streptomicin and ethambutol. Since the isolates did not grow at PNB (para-nitro benzoic acid) medium and niacin and nitrate activities were negative, spoligotyping (spacer oligonucleotide typing) was performed and DR loci characteristic for M. bovis BCG strain were detected. However, the patient died 2 weeks before the culture results were obtained. The effective use of mycobacteriology laboratories and cooperation between laboratory and clinics provide advantages in the early diagnosis and treatment of tuberculosis cases, decreasing the morbidity and the mortality.


Obstructive sleep apnoea in Singapore: polysomnography data from a tertiary sleep disorders unit.
Comprehensive sleep architecture and respiratory event data in local patients with suspected obstructive sleep apnoea (OSA) from overnight polysomnography (PSG), the gold standard for the evaluation of sleep-related breathing disorders, are not widely available. We present 1 year retrospective PSG data with the objective of describing PSG characteristics of patients evaluated for OSA in Singapore. PSG data of patients evaluated for OSA in 1 year (January through December 2005) in the Sleep Laboratory of a public tertiary hospital were retrospectively reviewed. Five hundred and eighty-four diagnostic PSG studies were performed in patients with symptoms suggestive of sleep-disordered breathing, including snoring, excessive daytime sleepiness, unrefreshing sleep, or recurrent unexplained awakenings. There were 449 male patients (76.9%) and 135 female patients (23.1%), with a mean age of 47.5 years (SD 12.7). Men were on average younger than women, 46.1 years versus 52.0 years (P <0.0005). The mean body mass index (BMI) was 27.9 (SD 6.7), with no significant difference between genders. An association was shown between apnoea-hypopnoea index (AHI) and BMI (Pearson correlation index r = 0.362). Men had overall significantly higher AHI (16.5 vs 9), shorter mean sleep onset latency (11 vs 16.5 minutes), more light sleep (65.5% vs 58.9%), less deep sleep (17.7% vs 23%), and more respiratory event related arousals per hour of sleep (11.6 vs 5.1) (P <0.0005). Severity was classified: AHI <5 ("Normal Overall AHI") (28.3%), AHI 5-15 ("Mild") (22.3%), AHI >15-30 ("Moderate") (18.3%), AHI >30 ("Severe") (31.2%). There was no significant age difference among the 4 groups. More severe OSA patients were significantly heavier, and had more light sleep, less deep sleep, less REM sleep, more respiratory event related arousals and lower levels of oxygen desaturation. OSA is predominant in middle-aged, overweight Singapore males and much less common in females who tend to be older. A majority of patients have moderate to severe OSA, which significantly disturbs normal sleep architecture. The relatively lower BMI compared to Caucasian OSA populations may be related to local craniofacial characteristics and/or higher percentage of body fat for BMI which has been described in Singaporeans.


Evidence for early domestic yak, taurine cattle, and their hybrids on the Tibetan Plateau.
Domestic yak, cattle, and their hybrids are fundamental to herder survival at high altitudes on the Tibetan Plateau. However, little is known about their history. Bos remains are uncommon in this region, and ancient domestic yak have not been securely identified. To identify Bos taxa and investigate their initial management, we conducted zooarchaeological analyses of 193 Bos specimens and sequenced five nuclear genomes from recently excavated assemblages at Bangga. Morphological data indicated that more cattle than yak were present. Ancient mitochondrial DNA and nuclear genome sequences identified taurine cattle and provided evidence for domestic yak and yak-cattle hybridization ~2500 years ago. Reliance on diverse Bos species and their hybrid has increased cattle adaptation and herder resilience to plateau conditions. Ancient cattle and yak at Bangga were closely related to contemporary livestock, indicating early herder legacies and the continuity of cattle and yak husbandry on the Tibetan Plateau.


Tempo and Mode in Cultural Macroevolution.
Evolutionary scientists studying social and cultural evolution have proposed a multitude of mechanisms by which cultural change can be effected. In this article we discuss two influential ideas from the theory of biological evolution that can inform this debate: the contrast between the micro- and macro-evolution, and the distinction between the tempo and mode of evolution. We add the empirical depth to these ideas by summarizing recent results from the analyses of data on past societies in Seshat: Global History Databank. Our review of these results suggests that the tempo (rates of change, including their acceleration and deceleration) of cultural macroevolution is characterized by periods of apparent stasis interspersed by rapid change. Furthermore, when we focus on large-scale changes in cultural traits of whole groups, the most important macroevolutionary mode involves inter-polity interactions, including competition and warfare, but also cultural exchange and selective imitation; mechanisms that are key components of cultural multilevel selection (CMLS) theory.


Prevalence of Streptococcus pneumoniae in respiratory samples from patients with tracheostomy in a long-term-care facility.
We aimed to study the prevalence of Streptococcus pneumoniae in respiratory samples from institutionalized patients with chronic tracheostomy. A total of 264 pairs of nasopharyngeal and endotracheal cultures were collected. There was no difference in the proportion of positive cultures between children (21%) and adults (18%). However, the proportion of positive endotracheal cultures was higher than that of nasopharyngeal cultures in adults (18% versus 3%, respectively; P < 0.001) but not in children (17% in both sites).


Determination of incoming solar radiation in major tree species in Turkey.
Light requirements and spatial distribution of major forest tree species in Turkey hasn't been analyzed yet. Continuous surface solar radiation data, especially at mountainous-forested areas, are needed to put forward this relationship between forest tree species and solar radiation. To achieve this, GIS-based modeling of solar radiation is one of the methods used in rangelands to estimate continuous surface solar radiation. Therefore, mean monthly and annual total global solar radiation maps of whole Turkey were computed spatially using GRASS GIS software "r.sun" model under clear-sky (cloudless) conditions. 147498 pure forest stand point-based data were used in the study for calculating mean global solar radiation values of all the major forest tree species of Turkey. Beech had the lowest annual mean total global solar radiation value of 1654.87 kWh m(-2), whereas juniper had the highest value of 1928.89 kWh m(-2). The rank order of tree species according to the mean monthly and annual total global solar radiation values, using a confidence level of p < 0.05, was as follows: Beech < Spruce < Fir species < Oak species < Scotch pine < Red pine < Cedar < Juniper. The monthly and annual solar radiation values of sites and light requirements of forest trees ranked similarly.


Epsilon-near-zero mode for active optoelectronic devices.
The electromagnetic modes of a GaAs quantum well between two AlGaAs barriers are studied. At the longitudinal optical phonon frequency, the system supports a phonon polariton mode confined in the thickness of the quantum well that we call epsilon-near-zero mode. This epsilon-near-zero mode can be resonantly excited through a grating resulting in a very large absorption localized in the single quantum well. We show that the reflectivity can be modulated by applying a voltage. This paves the way to a new class of active optoelectronic devices working in the midinfrared and far infrared at ambient temperature.


Synthetic and analytical strategies for the quantification of phenyl-γ-valerolactone conjugated metabolites in human urine.
The contribution of the gut microbiota to the metabolism of catechins and proanthocyanidins remains unclear. Phenyl-γ-valerolactones have been identified as the most representative metabolites of these dietary flavan-3-ols, but their accurate quantification has posed problems because of a lack of appropriate bioanalytical standards. This work aimed at synthesizing a novel set of sulphate- and glucuronide-conjugated phenyl-γ-valerolactones and at developing an analytical platform using UHPLC-ESI-MS/MS for their quantification in urine. Eight glucuronide and sulphate conjugates of hydroxyphenyl-γ-valerolactones were synthesized and used as analytical standards, together with five phenyl-γ-valerolactone aglycones, for the development of a high-throughput, validated analytical method. Chromatographic and MS conditions were optimized. The method validation showed acceptable linearity, intra-day and inter-day repeatability, and accuracy, with the analytical range, limit of detection (LOD), and lower limit of quantification (LLOQ) varying notably among compounds. The method was used to calculate the excretion of phenyl-γ-valerolactones in healthy subject consuming green tea, providing novel information on the real concentrations of phenyl-γ-valerolactones in urine. This work opens the door to better studying the bioavailability of flavan-3-ols and the real exposition to flavan-3-ol sources, as well as to define the bioactivity of these colonic metabolites in cell assays.


Ultrastructural studies on the intracellular fate of Chlamydia psittaci (strain guinea pig inclusion conjunctivitis) and Chlamydia trachomatis (strain lymphogranuloma venereum 434): modulation of intracellular events and relationship with endocytic mechanism.
Previous observations on the highly infectious LGV strain 434 of Chlamydia trachomatis and the guinea pig inclusion conjunctivitis (GPIC) strain of C. psittaci (which requires centrifugation of inocula with host cell monolayers for maximum infectivity) indicated that infectivity differences were expressed, not at entry, but at an intracellular stage affecting multiplication. Centrifugation increased the potential of internalized chlamydiae to undergo productive infection. Here, analysis of the intracellular fate of chlamydiae by ultrastructural methods indicates that strain GPIC exhibits two patterns of behaviour depending on the mode of inoculation. Strain GPIC showed limited entry, with 47% of intracellular organisms becoming associated with thorotrast-labelled lysosomes, following static incubation with monolayers. In contrast, with centrifugation, entry was not limited and association with lysosomes was reduced to 12%; strain 434 behaved similarly but independently of the mode of inoculation. The different results for strain GPIC correlated with distinct entry mechanisms. Entry during static incubation was unimpaired either by treatment with cytochalasin D or by temperature reduction to 20 degrees C, suggesting that it was pinocytic. Entry during centrifugation was markedly impaired by both treatments, suggesting that it was phagocytic. The data lead to two novel conclusions: first, that chlamydiae can apparently enter cells by both pinocytic and phagocytic mechanisms; second, that the entry mechanism influences intracellular fate. It is suggested that entry mechanism is linked to selection of the vesicle membrane forming around the internalizing chlamydiae. This, in turn, may influence both intracellular translocation and subsequent inhibition or promotion of multiplication of the internalized parasite.


Disparities in rates of acute MI hospitalization and coronary procedures on the US-Mexico border.
Hospitalization rates for acute myocardial infarction can provide insight into the utilization of care by disadvantaged populations. However, these data have not been reported for the US-Mexico border region. Hospital discharge public use data files for 2000 were obtained from the health departments of Arizona, California, and Texas. The diagnosis of acute myocardial infarction was based on International Classification of Diseases-9th Revision, Clinical Modification code 410 as a primary discharge diagnosis. In addition, cardiac catheterization, coronary angioplasty, and bypass grafting procedures were enumerated. Discharges were classified as occurring in hospitals in border counties and nonborder counties. Overall, 12,464 hospital discharges in border counties had acute myocardial infarction listed as the first diagnosis. Among those aged 45-64 and >or=65 years in border counties, Texas had the lowest discharge rates (eg, at >or=65 years: Texas 95, California 134 per 10,000), lower than in nonborder counties. Among those aged >or=65 years, rates in Texas also were lower in border than nonborder counties, a pattern not seen in other states. On the contrary, rates of catheterization and bypass grafting were highest in Texas, whereas the rates of bypass grafting were actually higher in border than in nonborder counties. In border and nonborder counties of the combined states, hospitalization rates of acute myocardial infarction in Hispanics were lower than those of non-Hispanics. Rates varied little between border and nonborder counties within ethnic groups. A similar pattern was observed for the hospital use of angioplasty and coronary bypass surgery. Our findings suggest possible underutilization of hospital in-patient care for coronary artery disease by Hispanics who were residents of Texas border counties. Further studies are needed to test this hypothesis using more recent data.


Safety and immunogenicity of an oral, replicating adenovirus serotype 4 vector vaccine for H5N1 influenza: a randomised, double-blind, placebo-controlled, phase 1 study.
Replication-competent virus vector vaccines might have advantages compared with non-replicating vector vaccines. We tested the safety and immunogenicity of an oral adenovirus serotype 4 vector vaccine candidate (Ad4-H5-Vtn) expressing the haemagglutinin from an avian influenza A H5N1 virus. We did this phase 1 study at four sites in the USA. We used a computer-generated randomisation list (block size eight, stratified by site) to assign healthy volunteers aged 18-40 years to receive one of five doses of Ad4-H5-Vtn (10(7) viral particles [VP], 10(8) VP, 10(9) VP, 10(10) VP, 10(11) VP) or placebo (3:1). Vaccine or placebo was given on three occasions, about 56 days apart. Participants, investigators, and study-site personnel were masked to assignment throughout the study. Subsequently, volunteers received a boost dose with 90 μg of an inactivated parenteral H5N1 vaccine. Primary immunogenicity endpoints were seroconversion by haemagglutination-inhibition (HAI), defined as a four-times rise compared with baseline titre, and HAI geometric mean titre (GMT). We solicited symptoms of reactogenicity daily for 7 days after each vaccination and recorded symptoms that persisted beyond 7 days as adverse events. Primary analysis was per protocol. This trial is registered with ClinicalTrials.gov, number NCT01006798. We enrolled 166 participants (125 vaccine; 41 placebo) between Oct 19, 2009, and Sept 9, 2010. HAI responses were low: 13 of 123 vaccinees (11%, 95% CI 6-17) and three of 41 placebo recipients (7%, 2-20) seroconverted. HAI GMT was 6 (95% CI 5-7) for vaccinees, and 5 (5-6) for placebo recipients. However, when inactivated H5N1 vaccine became available, one H5N1 boost was offered to all participants. In this substudy, HAI seroconversion occurred in 19 of 19 participants in the 10(11) VP cohort (100%; 95% CI 82-100) and eight of 22 placebo recipients (36%; 17-59); 17 of 19 participants in the 10(11) VP cohort (89%; 67-99) achieved seroprotection compared with four of 22 placebo recipients (18%; 5-40); GMT was 135 (89-205) with 10(11) VP, compared with 13 (7-21) with placebo. The cumulative frequency of abdominal pain, diarrhoea, and nasal congestion after all three vaccinations was significantly higher in vaccinees than placebo recipients (21 [16·8%] of 125 vs one [2·4%] of 41, p=0·017; 24 [19·2%] of 125 vs two [4·9%] of 41, p=0·027; 41 [32·8%] of 125 vs six [14·6%] of 41, p=0·028; respectively). No serious treatment-related adverse events occurred. Oral Ad4 vector priming might enhance the efficacy of poorly immunogenic vaccines such as H5N1. Wellcome Trust Foundation, PaxVax.


[MOPr-DOPr heteromer: the meaning and possibility as novel therapeutic target for pain control].
Many studies suggest opioid receptor (OPr) dimerization modulates the pharmacological properties of opiates. Specifically, heteromerization between OPr types has been reported to lead to changes in intracellular signaling. Thus, ligands targeting heteromers are expected to be novel therapeutic targets with reduced side effects. The heteromers of mu (MOPr) and delta (DOPr) are detected in brain regions involved in pain processing. The bivalent ligand or small molecule were identified as a MOPr-DOPr targeting ligand. These ligands exhibit antinociceptive properties similar to that of morphine with lesser antinociceptive tolerance as compared to morphine. Studies exploring the in vivo regulation of MOPr-DOPr heteromers, showed chronic morphine administration leads to an upregulation of these heteromers in select brain regions. Exploration of mechanisms underlying this phenomenon led us to the G protein-coupled receptor chaperone, RTP4, that is induced by chronic morphine and facilitates the heteromerization of MOPr and DOPr. In this review, I will introduce the simulated structure or property of MOPr-DOPr heteromer, its targeting ligands, and its intracellular regulatory mechanism that include a key molecule like RTP4 that could serve as a scaffold for the development of novel therapeutic drugs with reduced adverse effects, and hence may take place of the conventional clinical opioids.


[Comprehensive medical and ecological studies in Pryel'brussi].
Research works in high-mountainous areas of Caucasus and, in particular, in Prielbrusie was initiated by N.N. Sirotinin as far back as in 1929. From the beginning the works were done in expeditionary conditions, from 1973--in further organized permanent Elbrus medical and biological station (EMBS). At the present time EMBS is a component of International Center for astronomical, medical and ecological research that was reorganized in 1999. EMBS is a unique scientific subdivision that has no analogues, it is located on different heights of Elbrus Mountain and complex fundamental scientific problems may be solved here using contemporary scientific methods. EMBS includes two-floor laboratory building, thermobaro-chamber, and the center organized for the recovery of patients from ecologically unfavorable regions and increasing of the sportsmen work capacity. Today the new stage of the EMBS activity starts and therefore there is a time to summarize some results of the research works here. Such results of EMBS activity are the fundamental studies in the field of oxygen insufficiency, which reveal the destructive and constructive mechanisms of the hypoxic state development in organisms. Such investigations became the base for the implementation for the first time in the world of the new highly effective methods of hypoxytherapy aimed on the patient treatment, prophylaxis, resistance increasing and improvement of athletic results for the sportsmen. There were suggested and proved the concept of step-by-step adaptation to hypoxia. Hypoxic states developed under the extreme condition influences as well as during different diseases here were studied comprehensively on the different organism levels. The methods of mathematic simulation of hypoxic states with prediction of organism states were elaborated at EMBS too. All these works transformed hypoxia studies from the descriptive-experimental science in exact one.


[Kaposi sarcoma-associated herpesvirus; human herpesvirus 8].
The discovery of a new human herpesvirus in Kaposi's sarcoma tissues of AIDS patients has opened up new facts in virology and oncology. This herpesvirus was first descriptively named Kaposi's sarcoma-associated herpesvirus, but was recently renamed human herpesvirus 8. Human herpesvirus 8 DNA has been consequently found in all forms of Kaposi's sarcoma, suggesting that it might be involved in the pathogenesis of the disease. Additionally, human herpesvirus 8 can be detected in both malignant and benign lymphoproliferative diseases, such as body-cavity-based B-cell lymphomas and multicentric Castleman disease. The virus was also recently found in rare vascular tumors in patients with angiosarcoma of the face and angiolymphoid hyperplasia with eosinophilia. Although only a limited portion of the viral DNA has been sequenced, it has become evident that the new herpesvirus is equipped with genes that could confer oncogenic potential. The virus can now be cultured, providing the possibility for studies of viral replication and the mode of transmission, and also for the development of serologic tests.


Improving recruitment of older people to clinical trials: use of the cohort multiple randomised controlled trial design.
There is widespread evidence of under-recruitment of older people to research studies, notably randomised controlled trials of interventions. Study exclusion criteria, ethical dilemmas, patient preference, risk of bias and challenges for treatment comparisons are particular problems faced by researchers. This article describes how more widespread use of the cohort multiple randomised controlled trial (cmRCT) design in ageing research may help address many of these problems. The original key features of the cmRCT design are a large observational cohort of people with the condition of interest (e.g. frailty) with regular measurement of outcomes for the whole cohort. For each RCT eligible patients are identified and a random selection offered the trial intervention; their outcomes are compared with those eligible patients not offered the intervention. Relevant assents are obtained at baseline to enable future involvement in a range of potential trials. Where possible, the follow-up schedule is aligned with the key time points for assessment in future trials and includes the key baseline descriptors, and primary and secondary outcomes. The cmRCT approach also enables detailed observational and qualitative research for the chosen condition of interest, and might include the establishment of research biobanks to better align basic science, epidemiological, qualitative and clinical trial research.


Vacuolar transport in tobacco leaf epidermis cells involves a single route for soluble cargo and multiple routes for membrane cargo.
We tested if different classes of vacuolar cargo reach the vacuole via distinct mechanisms by interference at multiple steps along the transport route. We show that nucleotide-free mutants of low molecular weight GTPases, including Rab11, the Rab5 members Rha1 and Ara6, and the tonoplast-resident Rab7, caused induced secretion of both lytic and storage vacuolar cargo. In situ analysis in leaf epidermis cells indicates a sequential action of Rab11, Rab5, and Rab7 GTPases. Compared with Rab5 members, mutant Rab11 mediates an early transport defect interfering with the arrival of cargo at prevacuoles, while mutant Rab7 inhibits the final delivery to the vacuole and increases cargo levels in prevacuoles. In contrast with soluble cargo, membrane cargo may follow different routes. Tonoplast targeting of an α-TIP chimera was impaired by nucleotide-free Rha1, Ara6, and Rab7 similar to soluble cargo. By contrast, the tail-anchored tonoplast SNARE Vam3 shares only the Rab7-mediated vacuolar deposition step. The most marked difference was observed for the calcineurin binding protein CBL6, which was insensitive to all Rab mutants tested. Unlike soluble cargo, α-TIP and Vam3, CBL6 transport to the vacuole was COPII independent. The results indicate that soluble vacuolar proteins follow a single route to vacuoles, while membrane spanning proteins may use at least three different transport mechanisms.


Breast-feeding and human immunodeficiency virus infection: assessment of knowledge among clinicians in Kenya.
In Kenya, human immunodeficiency virus (HIV) prevalence ranks among the highest in the world. Approximately 60 000 infections yearly are attributed to vertical transmission including the process of labour and breast-feeding. The vast of the population affected is in the developing world. Clinical officers and nurses play an important role in provision of primary health care to antenatal and postnatal mothers. There are a few studies that have explored the clinicians' knowledge on breast-feeding in the face of HIV and in relation to vertical transmission this being a vital component in prevention of maternal-to-child transmission. The aim of this study was to evaluate clinicians' knowledge on HIV in relation to breast-feeding in Kenya. A cross-sectional survey was conducted to assess knowledge of 161 clinical officers and nurses serving in the maternity and children' wards in various hospitals in Kenya. The participants were derived from all district and provincial referral facilities in Kenya. A preformatted questionnaire containing a series of questions on HIV and breast-feeding was administered to clinicians who were then scored and analyzed. All the 161 participants responded. Majority of clinicians (92%) were knowledgeable regarding prevention of mother-to-child transmission. Regarding HIV and breast-feeding, 49.7% thought expressed breast milk from HIV-positive mothers should be heated before being given. Majority (78.3%) thought breast milk should be given regardless of availability of alternatives. According to 74.5% of the participants, exclusive breast-feeding increased chances of HIV transmission. Two-thirds (66.5%) would recommend breast-feeding for mothers who do not know their HIV status (66.5%). This study observes that a majority of the clinicians have inadequate knowledge on breast-feeding in the face of HIV. There is need to promote training programmes on breast-feeding and transmission of HIV from mother to child. This can be done as in-service training, continuous medical education and as part of the formal training within medical institutions.


[Unemployment, job insecurity and their consequences for health in a sample of young adults].
Negative consequences of unemployment have been known for a long time. This study concerns the effect of unemployment and job insecurity in a large sample of young adults because little is known about this special age group. Methods : 420 persons (46.8% males, 52.2% females, mean age 29 years) were polled in the 16(th) wave of the Sächsische Längsschnittstudie in 2002. This longitudinal study accompanies an East German sample since 1987 i.e. some time before German reunification. We used standardised psychological questionnaires to assess the state of health (SCL-9, HADS, GBB, SWE). 120 (29%) persons were repeatedly unemployed, 143 (34%) once, and only 157 (37%) have never been unemployed. The period of unemployment lasted 1 to 76 months. According to the experience with unemployment we found differences in subgroups: persons having more experience with unemployment report on higher global distress, more anxiety and depression, feel less efficacious and are in a subjectively poorer state of health. Nearly one-third of the participants think they have an insecure job. Persons who perceive an insecure job feel significantly greater anxiety, depression, body complaints, mental distress and feel less efficacious. Unemployment is a big social problem for young and well-qualified persons. The experience of unemployment decreases the identification with the current social system and has a strong negative influence on the state of health. Specific offers of medical and psychosocial support are required. Even the feeling of job insecurity has explicitly negative effects on health. Further longitudinal research is necessary.


Hexosome and hexagonal phases mediated by hydration and polymeric stabilizer.
In this research, we studied the factors that control formation of GMO/tricaprylin/water hexosomes and affect their inner structure. As a stabilizer of the soft particles dispersed in the aqueous phase, we used the hydrophilic nonionic triblock polymer Pluronic 127. We demonstrate how properties of the hexosomes, such as size, structure, and stability, can be tuned by their internal composition, polymer concentration, and processing conditions. The morphology and inner structure of the hexosomes were characterized by small-angle X-ray scattering, cryo-transmission electron microscope, and dynamic light scattering. The physical stability (to creaming, aggregation, and coalescence) of the hexosomes was further examined by the LUMiFuge technique. Two competing processes are presumed to take place during the formation of hexosomes: penetration of water from the continuous phase during dispersion, resulting in enhanced hydration of the head groups, and incorporation of the polymer chains into the hexosome structure while providing a stabilizing surface coating for the dispersed particles. Hydration is an essential stage in lyotropic liquid crystal (LLC) formation. The polymer, on the other hand, dehydrates the lipid heads, thereby introducing disorder into the LLC and reducing the domain size. Yet, a critical minimum polymer concentration is necessary in order to form stable nanosized hexosomes. These competing effects require the attention of those preparing hexosomes. The competition between these two processes can be controlled. At relatively high polymer concentrations (1-1.6 wt % of the total formulation of the soft particles), the hydration process seems to occur more rapidly than polymer adsorption. As a result, smaller and more stable soft particles with high symmetry were formed. On the other hand, when the polymer concentration is fixed at lower levels (<1.0 wt %), the homogenization process encourages only partial polymer adsorption during the dispersion process. This adsorption is insufficient; hence, maximum hydration of the surfactant head group is reached prior to obtaining full adsorption, resulting in the formation of less ordered hexosomes of larger size and lower stability.


A model for the phosphorylation of the Ca2+ + Mg2+-activated ATPase by phosphate.
We have shown that changes in fluorescence intensity for the Ca2+ + Mg2+-activated ATPase of sarcoplasmic reticulum labelled with fluorescein isothiocyanate following the addition of Ca2+ can give the ratio of the two conformations (E1 and E2) of the ATPase. We show that the fluorescence response to Ca2+ is unaffected by Mg2+, phosphate or K+, implying that these ions bind equally well to the E1 and E2 conformations. A model is presented for phosphorylation of the ATPase by phosphate as a function of pH, Mg2+, K+ and Ca2+.


Hamartomatous polyposis syndromes.
The hamartomatous polyposis syndromes are a heterogeneous group of disorders that share an autosomal-dominant pattern of inheritance and are characterized by hamartomatous polyps of the gastrointestinal tract. These syndromes include juvenile polyposis syndrome, Peutz-Jeghers syndrome and the PTEN hamartoma tumor syndrome. The frequency and location of the polyps vary considerably among syndromes, as does the affected patient's predisposition to the development of gastrointestinal and other malignancies. Although the syndromes are uncommon, it is important for the clinician to recognize these disorders because they are associated with considerable morbidity and mortality, not only from malignancy but also from nonmalignant manifestations such as bleeding, intussusception, and bowel obstruction. Each hamartomatous polyposis syndrome has its own distinctive organ-specific manifestations and each requires a different surveillance strategy, which makes accurate diagnosis crucial for appropriate patient management. The availability of clinical genetic testing for these disorders means that appropriate recognition allows for timely referral for cancer genetic counseling, and often allows for predicative testing in at-risk family members. Promisingly, an understanding of the molecular pathogenesis of these disorders offers insights into the mechanisms underlying the development of sporadic malignancy, and enables rational selection of targeted therapies that warrant further investigation.


Robust T cell responses to aspergillosis in chronic granulomatous disease: implications for immunotherapy.
Chronic granulomatous disease (CGD) patients are highly susceptible to invasive aspergillosis and might benefit from aspergillus-specific T cell immunotherapy, which has shown promise in treating those with known T cell defects such as haematopoietic stem cell transplant (HSCT) recipients. But whether such T cell defects contribute to increased risks for aspergillus infection in CGD is unclear. Hence, we set out to characterize the aspergillus-specific T cell response in CGD. In murine CGD models and in patients with CGD we showed that the CD4(+) T cell responses to aspergillus were unimpaired: aspergillus-specific T cell frequencies were even elevated in CGD mice (P < 0·01) and humans (P = 0·02), compared to their healthy counterparts. CD4-depleted murine models suggested that the role of T cells might be redundant because resistance to aspergillus infection was conserved in CD4(+) T cell-depleted mice, similar to wild-type animals. In contrast, mice depleted of neutrophils alone or neutrophils and CD4(+) T cells developed clinical and pathological evidence of pulmonary aspergillosis and increased mortality (P < 0·05 compared to non-depleted animals). Our findings that T cells in CGD have a robust aspergillus CD4(+) T cell response suggest that CD4(+) T cell-based immunotherapy for this disease is unlikely to be beneficial.


Protective immunity and safety of a genetically modified influenza virus vaccine.
Recombinant influenza viruses are promising viral platforms to be used as antigen delivery vectors. To this aim, one of the most promising approaches consists of generating recombinant viruses harboring partially truncated neuraminidase (NA) segments. To date, all studies have pointed to safety and usefulness of this viral platform. However, some aspects of the inflammatory and immune responses triggered by those recombinant viruses and their safety to immunocompromised hosts remained to be elucidated. In the present study, we generated a recombinant influenza virus harboring a truncated NA segment (vNA-Δ) and evaluated the innate and inflammatory responses and the safety of this recombinant virus in wild type or knock-out (KO) mice with impaired innate (Myd88 -/-) or acquired (RAG -/-) immune responses. Infection using truncated neuraminidase influenza virus was harmless regarding lung and systemic inflammatory response in wild type mice and was highly attenuated in KO mice. We also demonstrated that vNA-Δ infection does not induce unbalanced cytokine production that strongly contributes to lung damage in infected mice. In addition, the recombinant influenza virus was able to trigger both local and systemic virus-specific humoral and CD8+ T cellular immune responses which protected immunized mice against the challenge with a lethal dose of homologous A/PR8/34 influenza virus. Taken together, our findings suggest and reinforce the safety of using NA deleted influenza viruses as antigen delivery vectors against human or veterinary pathogens.


[Ileal ureteral substitute and ileal urinary diversion in conjunction with renal transplantation (author's transl)].
A patient with transplanted kidney and immunosuppression treatment has been treated because of a total necrosis of ureter by a replacement of a ureter with a pyeloileocystoplasty. A second patient with transplanted kidney and immunosuppression therapy has been given care to with total cystectomy and supravesical derivation of the urine by a ileal conduit of a solid cancer or urinary bladder. The good clinical results, five, respectively two years after the operation confirm that neither kidney transplantation nor immunosuppression therapy are counter indications to the use of small intestine plastics of the bladder of the ureter.


Comparative activity of piperacillin/tazobactam against 5625 isolates from hospitalised patients. Multicentre Study Group.
The comparative activities of piperacillin/tazobactam and other antibiotics against 5625 Gram-negative and Gram-positive bacteria freshly isolated from hospitalized patients was examined. The percentage of Enterobacteriaceae susceptible to piperacillin/tazobactam, ceftazidime, imipenem and ciprofloxacin was very similar and varied from 81% to 84% in the Enterobacteriaceae with inducible type 1 beta-lactamases and from 93 to 97% in the non-inducible organisms. Piperacillin/tazobactam was similar in activity to ceftazidime against Pseudomonas aeruginosa and more active than imipenem or ciprofloxacin. Piperacillin/tazobactam is more active than ceftazidime against Gram-positive bacteria (staphylococci, enterococci) and seems to be a promising antibiotic for nosocomial infections.


Innovation focus: the patient with arrhythmia.
Great strides have been made over the last two decades in the management of patients with rhythm disorders. Despite this, however, the remaining critical problems of stroke related to atrial fibrillation or as a result of radiofrequency ablation require innovative solutions to fully realize the potential of these recent advances. Similarly, implanted cardiac devices have revolutionized the care of patients with bradyrhythmias and tachyarrhythmias. Dyssynchronus ventricular pacing associated with present devices; however, results in heart failure, tricuspid regurgitation, and inappropriate device therapy once again create a demand for creative solutions. While not technically an arrhythmia, epilepsy management today is riddled with many of the problems that plagued cardiac arrhythmia management previously, and thus an appreciation of the similarities in requirement for investigative solutions may yield groundbreaking solutions. In this paper, we describe some novel methods to reduce complications associated with rhythm disorders and their treatment and apply the lessons learned from cardiovascular arrhythmia management to the brain. These include: a method to reduce coagulum formation and thus subsequent thromboembolism with indwelling catheters specifically during radiofrequency ablation procedures; a technique to ligate the left atrial appendage through percutaneous subxiphoid pericardial access; development and testing of a novel intramyocardial pace-sense lead, particularly used in a unique anatomic location (the atrioventricular septum) to allow pacing the ventricles in a relatively synchronous manner without crossing the tricuspid valve or entering the coronary sinus; finally, novel modifications of the cardiovascular mapping and ablation techniques used for the management of the central nervous system disorders primarily via the venous drainage of the brain. Innovative and potential solutions to treat the patient with arrhythmia are presented.


Thermospray HPLC/MS: a new mass spectrometric technique for the profiling of steroids.
The analysis of various steroid classes by thermospray HPLC-MS using solvent systems containing 0.1 M ammonium acetate has been described. For simple unconjugated 3-oxo-4-ene steroids the positive ion spectra are dominated by a parent ion M + H+ and with increasing numbers of hydroxyl group intense ions formed by sequential losses of water (M + H- n18)+ become important. Steroids with dihydroxyacetone side-chains readily lose these side-chains and the resulting (M + H-60)+ fragment is the base peak in their spectra. The (M + H-60)+ ion is not important for most steroids with glycerol-type side-chains. Although competition between thermal degradation and vaporization was observed at lower concentrations, the effect was minimized after optimizing conditions and the protonated molecular ion was easily detected when as little as 1-10 pmol of material were injected on-column. Steroid glucuronides when analyzed in the negative ion mode give simple spectra with base peak and parent ion (M-H)-. Lack of fragmentation permits facile and sensitive measurement of individual glucoronides by selected-ion-monitoring. Extensive fragmentation is seen in the positive ion mode with sequential losses of H2O from the molecular ions (M + NH4)+ and from the aglycone fragment ion. For simple unconjugated steroids the sensitivity of HPLC-MS in selected-ion-monitoring mode can be excellent. When the protonated molecular ion of testosterone was monitored the signal/noise ratio for 30 pg testosterone was about 10.


The comparison of fluid dynamics parameters in an Andersen cascade impactor equipped with and without a preseparator.
The fluid dynamic data in Andersen cascade impactor (ACI) are still lacking. Airflows and those affected parameters can be predicted in a preseparator and Andersen cascade impactor (ACI) by computational modeling. This study developed a validated computational fluid dynamic (CFD) model of an ACI and investigated the effects of the preseparator on the CFD parameters. Validation of the computational nozzle velocity for each of the stage 0 to stage 5 of the ACI stages was found to be within a 3.56% error. The flow field indicated that the preseparator accelerated the airflow velocity at the induction tube from 1.13 to 3.71 ± 0.09 m/s and 2.40 to 8.68 ± 0.16 m/s (at 28.3 and 60 L/min of flow rate, respectively). The preseparator produced a nozzle's wall shear stress ranged from 0.08 to 0.34 Pa on a collection plate, while the ex-preseparator spread wall shear from the plate's center was in a range of 0.11 to 0.37 Pa (at 28.3 L/min of flow rate). Moreover, the nozzle velocities increased along the distance from the middle of the collection plate to the periphery. The CFD explained the airflow of the preseparator equipped model by accelerating the airflow along the inlet port to maximize the trapping of desirable particles and the generation of a smooth wall shear stress at the collection plate to reduce the particle re-entrainment. While, the ex-preseparator generated an airflow that resulted in a higher wall shear stress occurring on the lower stages.


Long-term mortality and quality of life after septic shock: a follow-up observational study.
In septic shock, short-term outcomes are frequently reported, while long-term outcomes are not. The aim of this study was to evaluate mortality and health-related quality of life (HRQOL) in survivors 6 months after an episode of septic shock. This single-centre observational study was conducted in an intensive care unit in a university hospital. All patients with septic shock were included. Mortality was assessed 6 months after the onset of septic shock, and a comparison between patients who survived and those who died was performed. HRQOL was assessed using the MOS SF-36 questionnaire prior to hospital admission (baseline) and at 6 months in survivors. HRQOL at baseline and at 6 months were compared to the general French population, and HRQOL at baseline was compared to 6-month HRQOL. Ninety-six patients were included. Six-month mortality was 45%. Survivors were significantly younger, had significantly lower lactate levels and SAPS II scores, required less renal support, received less frequent administration of corticosteroids, and had a longer length of hospital stay. At baseline (n = 39) and 6 months (n = 46), all of the components of the SF-36 questionnaire were significantly lower than those in the general population. Compared to baseline (n = 23), the Physical Component Score (CS) improved significantly at 6 months, the Mental CS did not differ. Mortality 6 months after septic shock was high. HRQOL at baseline was impaired when compared to that of the general population. Although improvements were noted at 6 months, HRQOL remained lower than that in the general population.


Phospholipase A2 augments contraction and intracellular calcium mobilization through thromboxane A2 in bovine tracheal smooth muscle.
Phospholipase A2 (PLA2) induces hyper-sensitivity to muscarinic agonists in airway smooth muscle in vitro. The precise mechanism of this is unknown, but might involve altered calcium homeostasis. In order to elucidate the effects of PLA2, on bovine tracheal smooth muscle contraction, isometric tension and intracellular calcium concentration ([Ca2+]i) were simultaneously measured in fura 2-loaded muscle strips. A high concentration of PLA2 (0.5 microg x mL(-1)) caused the muscle strips to contract, and this contractile response was significantly attenuated by pretreatment with indomethacin (IND; 10 microM), but not by nordihydroguaiaretic acid (NDGA; 10 microM). A low concentration of PLA2 (0.02 microg x mL(-1)) did not directly contract muscle strips. However a low concentration PLA2 significantly enhanced the threshold of the contractile response and that of the [Ca2+]i response to acetylcholine (ACh), but not that of the response to a high K+ concentration. These augmented responses to ACh returned to control levels after pretreatment with IND, a thromboxane (TX) synthetase inhibitor (OKY-046; 10 microM) or a TXA2 receptor antagonist (ONO-3708; 10 microM), but not after NDGA pretreatment. These results suggest that a low concentration of phospholipase A2 enhances smooth muscle responsiveness to acetylcholine by agonist-mediated Ca2+ mobilization facilitated by thromboxane A2. It is concluded that phospholipase A2 plays an important role in bronchial hypersensitivity involving thromboxane A2. It remains to be examined whether similar abnormalities in calcium homeostasis and muscarinic receptor function or coupling are involved in the pathogenesis of asthma.


Crystal structure of Glycine max glutathione transferase in complex with glutathione: investigation of the mechanism operating by the Tau class glutathione transferases.
Cytosolic GSTs (glutathione transferases) are a multifunctional group of enzymes widely distributed in Nature and involved in cellular detoxification processes. The three-dimensional structure of GmGSTU4-4 (Glycine max GST Tau 4-4) complexed with GSH was determined by the molecular replacement method at 2.7 A (1 A=0.1 nm) resolution. The bound GSH is located in a region formed by the beginning of alpha-helices H1, H2 and H3 in the N-terminal domain of the enzyme. Significant differences in the G-site (GSH-binding site) as compared with the structure determined in complex with Nb-GSH [S-(p-nitrobenzyl)-glutathione] were found. These differences were identified in the hydrogen-bonding and electrostatic interaction pattern and, consequently, GSH was found bound in two different conformations. In one subunit, the enzyme forms a complex with the ionized form of GSH, whereas in the other subunit it can form a complex with the non-ionized form. However, only the ionized form of GSH may form a productive and catalytically competent complex. Furthermore, a comparison of the GSH-bound structure with the Nb-GSH-bound structure shows a significant movement of the upper part of alpha-helix H4 and the C-terminal. This indicates an intrasubunit modulation between the G-site and the H-site (electrophile-binding site), suggesting that the enzyme recognizes the xenobiotic substrates by an induced-fit mechanism. The reorganization of Arg111 and Tyr107 upon xenobiotic substrate binding appears to govern the intrasubunit structural communication between the G- and H-site and the binding of GSH. The structural observations were further verified by steady-state kinetic analysis and site-directed mutagenesis studies.


Amino acid metabolism after lambda-carrageenan injury to rat skeletal muscle.
To evaluate whether the alterations in amino acid (AA) metabolism that follow the lambda-carrageenan injury are direct consequences of wounding and are therefore independent of food intake, plasma AA, in vivo muscle intracellular free AA, and AA release during isolated hindlimb perfusions were determined in pentobarbital-anesthetized rats with lambda-carrageenan-induced hindlimb muscle wounds (W) and in pair-fed (PFC) and ad libitum-fed (ALC) non-wounded controls. Both PFC and W animals showed different plasma and muscle intracellular AA composition that ALC. The alterations observed in these compartments in W animals may not have resulted exclusively as a consequence of the wound. Wounded hindlimbs released more AA during perfusion than either control group. The marked increase in net protein catabolism of W muscle appeared to be related to wounding and not a consequence of partial starvation. The normalized release (amino acid/phenylalanine ratio) of glutamine (W less than ALC), alanine (W less than PFC and ALC), and the branched-chain amino acids (W greater than PFC and ALC) differed among groups, suggesting an alteration in the metabolism of these AA in W tissue.


Gasoline Vapor Emissions During Vehicle Refueling Events in a Vehicle Fleet Saturated With Onboard Refueling Vapor Recovery Systems: Need for an Exposure Assessment.
Background: Gasoline contains large proportions of harmful chemicals, which can be released during vehicle refueling. Onboard Refueling Vapor Recovery (ORVR) can reduce these emissions, but there is limited research on the system's efficacy over time in an actual vehicle fleet. The aims of this study are: (1) determine the feasibility of using an infrared camera to view vapor emissions from refueling; (2) examine the magnitude of refueling-related emissions in an ORVR-saturated fleet, to determine need for an exposure-assessment. Methods: Using an infrared camera optimized for optical gas imaging of volatile organic chemicals, refueling was recorded for 16 vehicles at six gas stations. Pumps were inspected for damage, refueling shut-off valve functioning, and presence of Stage II Vapor Recovery. Vehicle make/model and age were recorded or estimated. Results: Vapor emissions were observed for 14 of 16 vehicles at each station, with severity varying substantially by vehicle make/model and age. Use of an infrared camera allowed for identification of vapor sources and timing of release, and for visualizing vapor trajectories. Discussion: Notably emissions occurred not only at the beginning and end of refueling but also throughout, in contrast to a prior study which did not detect increases in atmospheric hydrocarbon levels mid-refueling. Future studies are vitally needed to determine the risk to individuals during typical refueling in an ORVR saturated vehicle fleet. We recommend comprehensive exposure-assessment including real-time monitoring of emitted volatile organic compounds paired with infrared gas-imaging and measurement of internal dose and health effects of gas station customers.


Inhalation toxicity of propineb. Part I: Results of subacute inhalation exposure studies in rats.
This article addresses results from repeated 1- and 4-wk inhalation exposure studies in Wistar rats with solid aerosol (dust) atmospheres of propineb, a zinc bisdithiocarbamate homopolymer that is used as an agrochemical fungicide. Groups of 10 rats/sex were exposed nose-only to mean concentrations of 3.97, 11.24, and 21.95 mg propineb/m(3) using an exposure regimen of 6 h/day, 5 days/wk for 4 wk. Concentrations were selected based on results from a pilot study in which rats were exposed under identical conditions on 5 consecutive days for 6 h/day to mean concentrations of 10.1, 19.9, 38.1, and 78.7 mg/m(3). Both studies demonstrated that with respect to muscular effects female rats were remarkably more susceptible as compared to males. Female rats exposed to 11.24 mg/m(3) and above displayed characteristic signs of toxicity that included weakness and flaccid paralysis of hindlegs and ensuing immobilization that was considered to be the cause of emaciation and ensuing mortality in some rats. There was an apparent reciprocal relationship of concentration and the manifestation of clinical evidence of muscular dysfunction; that is, the onset in female rats exposed to 11.24, 21.95, 38.1, and 78.7 mg/m(3) was on days 15, 8, 4, and 3, respectively. In contrast, none of the male rats elaborated comparable effects up to 38.1 mg/m(3). Neuromuscular measures included leg grip strength and supplemented the clinical findings, whereas the landing foot splay was only minimally affected. Hematology and clinical pathology endpoints, including those addressing thyroidal function, were unobtrusive up to and including 78.7 mg/m(3). Lung weights were significantly increased in groups exposed to 21.95 mg/m(3) and above, especially in male rats. The microscopic examinations made in the 4-wk study demonstrated an increased incidence of intraalveolar material and enlarged, foamy alveolar macrophages at 3.97 mg/m(3) and above. Especially in female rats an atrophy of thigh muscle fibers, including increased nuclei and focal degeneration, occurred at 11.24 mg/m(3) and above. TTCA (2-thiazolidinethione-4-carboxylic acid) in urine, a metabolite and biomarker of exposure to CS(2), which is a putative breakdown product of propineb, was proportionally higher in the female rats exposed to 11.24 mg/m(3) and above. This biomarker appears to accumulate with time. This finding provides indirect evidence that the etiopathologic cause of neuromuscular changes is related to intermediary levels of CS(2). The data of this investigation suggest that the toxicity of inhaled propineb is characterized by two independent effects, namely, responses occurring at the alveolar level and muscular weakness, especially in female rats. With respect to the latter finding, the no-observed-adverse-effect level (NOAEL) of the 4-wk study is 3.97 mg/m(3). Further study is needed to clarify whether the pulmonary response observed at this exposure level is consistent with an adaptive or an early adverse effect.


Osteoarthritis year 2012 in review: biomarkers.
Biomarkers provide useful diagnostic information by detecting cartilage degradation in osteoarthritis (OA), reflecting disease-relevant biological activity and predicting the course of disease progression. They also serve as surrogate endpoints in the drug discovery process. The aim of this narrative review was to focus on OA biomarker-related papers published between the osteoarthritis research society international (OARSI) 2011 meeting in San Diego and the OARSI 2012 meeting in Barcelona. The PubMed/MEDLINE and SciVerse Scopus bibliographic databases were searched using the keywords: 'biomarker' and 'osteoarthritis' and/or 'biomarker' and 'proteomics'. Ninety-eight papers were found with the keywords 'biomarker' and 'osteoarthritis'. Fifteen papers were found with the keywords 'biomarker' and 'proteomics'. Review articles were also included. The most relevant published studies focused on extracellular matrix (ECM) molecules in body fluids. Enrichment of the deamidated epitope of cartilage oligomeric matrix protein (D-COMP) suggests that OA disease progression is associated with post-translational modifications that may show specificity for particular joint sites. Fibulin-3 peptides (Fib3-1 and Fib3-2) have been proposed as potential biomarkers of OA along with follistatin-like protein 1 (FSTL1), a new serum biomarker with the capacity to reflect the severity of joint damage. The 'membrane attack complex' (MAC) component of complement has also been implicated in OA. Novel OA biomarkers are needed for sub-clinical disease diagnosis. Proteomic techniques are beginning to yield useful data and deliver new OA biomarkers in serum and urine. Combining biochemical markers with tissue and cell imaging techniques and bioinformatics (i.e., machine learning, clustering, data visualization) may facilitate the development of biomarker combinations enabling earlier detection of OA.


Isolation and Characterization of Neisseria meningitidis Groups A, C, X, and Y Polysaccharide Antigens.
To prepare Neisseria meningitidis groups A, C, X, and Y polysaccharide antigens, culture supernatant fluids were subjected to serial processes of salt precipitation, alkaline hydrolysis, ethyl alcohol precipitation, and Sephadex G-200 chromatography. This method resulted in the isolation of large quantities of group antigens. All are acidic polysaccharides, the group C antigen being a polymer of n-acetyl neuraminic acid. Thiobarbituric acid assay failed to reveal sialic acids in the other group antigens. Protein was undetectable by absorption at 280 nm or by Folin analysis. These antigens are of similar molecular size, the majority of which are excluded by Sephadex G-200. They migrate in the upper one-third of sucrose density gradients and are retained by 5% acrylamide gel. All are highly group-specific and react only with homologous hyperimmune antisera in hemagglutination, complement fixation, and immunodiffusion systems. As little as 0.03 mumoles of n-acetyl neuraminic acid in group C antigen inhibits the hemagglutination of group C-sensitized red cells. All antigens are immunogenic in rabbits. These techniques afford a simplified method for the production of relatively large yields of highly specific group antigens which participate in multiple immunologic systems.


The relationship between nurses sociodemographic characteristics and stressor life events.
To determine how nurses who encounter stressful live events adapt themselves, and to examine the relation between their personal characteristics and stressful events. The cross-sectional study was conducted between February and April 2007, and comprised all nurses working at the two public-sector hospitals in Manisa, Turkey. Data was collected using a pre-tested questionnaire. Data was analysed using SPSS version 11.0. Of the total 87 nurses, 33 (37.9%) were in the 28-32 age group; 67 (77.0%) were married; 35 (40.2%) had a child; 61 (70.1%) had an income equivalent to their expenses; 44 (50.6%) had an associate degree; 21 (24.22%) had 10-12 years of experience in the profession; 76 (87.4%) were working as service nurses, and 31 (35.6%) had worked for 1-3 years in this service. There was a relationship between some of the socio-demographic characteristics of the nurses and stressor life events.