Conformational risk factors of brachycephalic obstructive airway syndrome (BOAS) in pugs, French bulldogs, and bulldogs.
Extremely brachycephalic, or short-muzzled, dog breeds such as pugs, French bulldogs, and bulldogs are prone to the conformation-related respiratory disorder-brachycephalic obstructive airway syndrome (BOAS). Affected dogs present with a wide range of clinical signs from snoring and exercise intolerance, to life-threatening events such as syncope. In this study, conformational risk factors for BOAS that could potentially aid in breeding away from BOAS were sought. Six hundred and four pugs, French bulldogs, and bulldogs were included in the study. Soft tape measurements of the head and body were used and the inter-observer reproducibility was evaluated. Breed-specific models were developed to assess the associations between the conformational factors and BOAS status based on functional grading. The models were further validated by means of a BOAS index, which is an objective measurement of respiratory function using whole-body barometric plethysmography. The final models have good predictive power for discriminating BOAS (-) and BOAS (+) phenotypes indicated by the area under the curve values of >80% on the receiver operating curves. When other factors were controlled, stenotic nostrils were associated with BOAS in all three breeds; pugs and bulldogs with higher body condition scores (BCS) had a higher risk of developing BOAS. Among the standardized conformational measurements (i.e. craniofacial ratio (CFR), eye width ratio (EWR), skull index (SI), neck girth ratio (NGR), and neck length ratio (NLR)), for pugs EWR and SI, for French bulldogs NGR and NLR, and for bulldogs SI and NGR showed significant associations with BOAS status. However, the NGR in bulldogs was the only significant predictor that also had satisfactory inter-observer reproducibility. A NGR higher than 0.71 in male bulldogs was predictive of BOAS with approximately 70% sensitivity and specificity. In conclusion, stenotic nostrils, BCS, and NGR were found to be valid, easily applicable predictors for BOAS (+).


Differential effects of steroid hormones on parameters of cell growth.
Steroid hormones affect the growth of many tumor cells both in vivo and in vitro. Growth of cells in vitro can be studied as either anchorage dependent (monolayer culture) or anchorage independent (suspension culture), and in each case, steroid hormones can alter log-phase proliferation rate, saturation density, or cell morphology. Results presented here demonstrate that different steroid hormones can have different effects on each of these parameters in breast cancer cells. In androgen-responsive cells, glucocorticoid and androgen both stimulated fibroblastic morphology and saturation density in monolayer and growth in suspension, but glucocorticoid inhibited log-phase proliferation rate while androgen stimulated it. By transfection experiments, it has been possible to separate the androgen-regulated stimulation of cell proliferation from the androgen-regulated cell morphology changes indicating that the two parameters are not totally interdependent. The same separation, however, was not achieved for glucocorticoid regulation. Further separation of hormone effects was achieved also during the natural course of progression to steroid autonomy. In particular, the androgen and glucocorticoid stimulations of growth in suspension were separated. The experiments described here carry important messages for the design and interpretation of any experiment aimed at elucidating molecular events involved in steroid-mediated cell growth in culture.


Metabolic flexibility of a prospective bioremediator: Desulfitobacterium hafniense Y51 challenged in chemostats.
Desulfitobacterium hafniense Y51 has been widely used in investigations of perchloroethylene (PCE) biodegradation, but limited information exists on its other physiological capabilities. We investigated how D. hafniense Y51 confronts the debilitating limitations of not having enough electron donor (lactate), or electron acceptor (fumarate) during cultivation in chemostats. The residual concentrations of the substrates supplied in excess were much lower than expected. Transcriptomics, proteomics and fluxomics were integrated to investigate how this phenomenon was regulated. Through diverse regulation at both transcriptional and translational levels, strain Y51 turned to fermenting the excess lactate and disproportionating the excess fumarate under fumarate- and lactate-limiting conditions respectively. Genes and proteins related to the utilization of a variety of alternative electron donors and acceptors absent from the medium were induced, apparently involving the Wood-Ljungdahl pathway. Through this metabolic flexibility, D. hafniense Y51 may be able to switch between different metabolic capabilities under limiting conditions.


Accelerated cardiomyopathy in maternally inherited diabetes and deafness.
The clinical features and course of cardiac involvement in a patient with maternally inherited diabetes and deafness associated with the mitochondrial DNA 3243 mutation are reported. A 45-year-old woman with maternally transmitted diabetes mellitus and deafness presented with congestive heart failure. The patient showed a short P-R interval on electrocardiogram (ECG) and had developed progression from left ventricular hypertrophy to a hypokinetic cardiomyopathy pattern over the course of 10 months. Rapid cardiac change was accompanied by left ventricular remodeling, as shown by wall thinning on echocardiogram and decrease in QRS voltages on ECG. Coronary arteriography revealed no significant stenosis. In the endomyocardial biopsy specimens, light microscopy showed nonspecific cardiomyopathic changes. Genetic testing for mitochondrial DNA mutations in peripheral blood lymphocytes revealed an adenine (A)-to-guanine (G) substitution at nucleotide 3243 in the mitochondrial DNA encoding the transfer RNA for leucine (tRNA Leu (UUR)). The proportion of mutant mitochondrial DNA was 25%. Two of the patient's daughters, aged 13 and 21 years, who were symptom free, were found to carry the same point mutation. A short P-R interval on ECG in the younger of them was the sole manifestation of the mutation. Unfortunately, 6 months after diagnosis, the patient died suddenly at home. Accelerated cardiomyopathy can occur as a mitochondria-related complication in patients with maternally inherited diabetes and deafness associated with the 3243 mutation.


Like fibrin, (DD)E, the major degradation product of crosslinked fibrin, protects plasmin from inhibition by alpha2-antiplasmin.
Plasmin generation is localized to the fibrin surface because tissue-type plasminogen activator (t-PA) and plasminogen bind to fibrin, an interaction that stimulates plasminogen activation over a hundred-fold. To ensure efficient fibrinolysis, plasmin bound to fibrin is protected from inhibition by alpha2-antiplasmin. (DD)E, a major soluble degradation product of cross-linked fibrin that is a potent stimulator of t-PA, compromises the fibrin-specificity of t-PA by promoting systemic activation of plasminogen. In this study we investigated whether (DD)E also protects plasmin from inhibition by alpha2-antiplasmin, facilitating degradation of this soluble t-PA effector. (DD)E and fibrin reduce the rate of plasmin inhibition by alpha2-antiplasmin by 5- and 10-fold, respectively. Kringle-dependent binding of plasmin to (DD)E and fibrin, with Kd values of 52 and 410 nM, respectively, contributes to the protective effect. When (DD)E is extensively degraded by plasmin, yielding uncomplexed fragment E and (DD), protection of plasmin from inhibition by alpha2-antiplasmin is attenuated. These studies indicate that (DD)E-bound plasmin, whose generation reflects the ability of (DD)E to stimulate plasminogen activation by t-PA, has the capacity to degrade (DD)E by virtue of its resistance to inhibition. This provides a mechanism to limit the concentration of (DD)E and maintain the fibrin-specificity of t-PA.


Management of obstructive sleep apnea in pediatric craniofacial anomalies.
Obstructive sleep apnea (OSA) is often associated with congenital craniofacial malformations such as Pierre-Robin Syndrome, Hemifacial Microsomia, Treacher Collins Syndrome resulting in decreased pharyngeal airway, which, in severe cases, leads to tracheostomy dependence. Some pediatric patients had tracheostomies done and others with severe respiratory distress were considered tracheostomy candidates. Twelve patients with severe respiratory distress without tracheostomy and ten patients with tracheostomy were treated by mandibular distraction osteogenesis using either external or internal devices. The expansion of mandibular framework was analyzed using bony cephalometric landmarks and computed tomography (CT). The results demonstrated average mandibular elongation of 29 mm on each side using the external devices and 22 mm using the internal devices, and an increase in mandibular volume and pharyngeal airway. The group of patients with tracheostomies were decannulated and in the patients with respiratory distress there was improved airway with improvement of signs and symptoms of OSA with elimination of oxygen requirement. Mandibular distraction is a useful method in younger children with OSA expanding the mandible and concomitantly advancing the base of tongue and hyoid bone increasing the pharyngeal airway. The external devices permit greater distraction length, the removal is simple but the devices are uncomfortable for the patients. On the other hand, the internal devices are more comfortable for patients but permit shorter distraction length and require a second operation for removal.


Bridging the gap in life expectancy of the aborigines in Taiwan.
Similar to the general population in Taiwan, the health of aborigines has steadily improved over the last 30 years, but the gap remains wide, especially in males, despite an infusion of substantial medical resources. The objectives of this study are to quantify the contribution of major causes of death to the gap in life expectancy and to propose initiatives to bridge the health gap between aborigines and the general population. This study included residents (slightly over 200000) from 30 'aboriginal townships' in Taiwan. The gap in life expectancy between aborigines and the general population was analysed by decomposing these gaps according to major causes of deaths. This analysis quantifies the contribution of different causes of deaths to the gap in life expectancy between the two populations. The overall mortality of aborigines in these townships was approximately 70% higher than the respective male and female general populations over the past 30 years. Mortality from infectious disease, cirrhosis of the liver, accidents, and suicide are substantially higher than the general population. The gap in life expectancy at birth in males was 8.5 years during 1971-1973, increasing to 13.5 years by 1998-2000, however, the gap in females remained relatively stable (8.0 years and 8.4 years, respectively). Of the 13.5-year difference in life expectancy in males, the differential mortality from diseases of the digestive system (mainly due to cirrhosis of the liver), accidents (from both motor vehicle and non-motor vehicle accidents), and infectious and parasitic disease contributed half (50%) of the gap in life expectancy. In females, the above primarily preventable causes of deaths accounted for 41% of the life expectancy gap. Based on the findings of this study, we suggest that future focus should be in the area of primary prevention in order to reduce the incidence of infectious and parasitic diseases, liver cirrhosis, and accidents.


Racial disparities in diabetes care processes, outcomes, and treatment intensity.
Black Americans with diabetes have a higher burden of illness and mortality than do white Americans. However, the extent to which differences in medical care processes and treatment intensity contribute to poorer diabetes outcomes is unknown. To assess racial disparities in the quality of diabetes care processes, intermediate outcomes, and treatment intensity. We conducted an observational study of 801 white and 115 black patients who completed the Diabetes Quality Improvement Project survey (response rate=72%) in 21 Veterans Affairs (VA) facilities using survey data; medical record information on receipt of diabetes services (A1c, low-density lipoprotein [LDL], nephropathy screen, and foot and dilated eye examinations), and intermediate outcomes (glucose control measured by A1c; cholesterol control measured by LDL; and achieved level of blood pressure); and pharmacy data on filled prescriptions. There were no racial differences in receipt of an A1c test or foot examination. Blacks were less likely than whites to have LDL checked in the past 2 years (72% vs. 80%, P<0.05) and to have a dilated eye examination (50% vs. 63%, P<0.01). Even after adjusting for patients' age, education, income, insulin use, diabetes self-management, duration, severity, comorbidities, and health services utilization, racial disparities in receipt of an LDL test and eye examination persisted. After taking into account the nested structure of the data using a random intercepts model, blacks remained significantly less likely to have LDL testing than whites who received care within the same facility (68% vs. 83%, P<0.01). In contrast, there were no longer differences in receipt of eye examinations, suggesting that black patients were more likely to be receiving care at facilities with overall lower rates of eye examinations. After adjusting for patient characteristics and facility effects, black patients were substantially more likely than whites to have poor cholesterol control (LDL > or =130) and blood pressure control (BP > or =140/90 mm Hg) (P<0.01). Among those with poor blood pressure and lipid control, blacks received as intensive treatment as whites for these conditions. We found racial disparities in some diabetes care process and intermediate outcome quality measures, but not in intensity of treatment for those patients with poor control. Disparities in receipt of eye examinations were the result of black patients being more likely to receive care at lower-performing facilities, whereas for other quality measures, racial disparities within facilities were substantial.


VivaxGEN: An open access platform for comparative analysis of short tandem repeat genotyping data in Plasmodium vivax populations.
The control and elimination of Plasmodium vivax will require a better understanding of its transmission dynamics, through the application of genotyping and population genetics analyses. This paper describes VivaxGEN (http://vivaxgen.menzies.edu.au), a web-based platform that has been developed to support P. vivax short tandem repeat data sharing and comparative analyses. The VivaxGEN platform provides a repository for raw data generated by capillary electrophoresis (FSA files), with fragment analysis and standardized allele calling tools. The query system of the platform enables users to filter, select and differentiate samples and alleles based on their specified criteria. Key population genetic analyses are supported including measures of population differentiation (FST), expected heterozygosity (HE), linkage disequilibrium (IAS), neighbor-joining analysis and Principal Coordinate Analysis. Datasets can also be formatted and exported for application in commonly used population genetic software including GENEPOP, Arlequin and STRUCTURE. To date, data from 10 countries, including 5 publicly available data sets have been shared with VivaxGEN. VivaxGEN is well placed to facilitate regional overviews of P. vivax transmission dynamics in different endemic settings and capable to be adapted for similar genetic studies of P. falciparum and other organisms.


Effects of motor fatigue on cortical activation level and functional connectivity during upper limb resistance training.
This study aimed to clarify the effects of motor fatigue on cortical activation levels and functional connectivity during upper limb resistance training using functional near-infrared spectroscopy (fNIRS). Ten healthy college students participated in a high intensity upper limb resistance training and fNIRS was used to measure the changes of oxyhemoglobin concentration changes (HbO) in bilateral sensorimotor cortex (SMC), premotor cortex (PMC), supplementary motor area (SMA), and dorsolateral prefrontal cortex (DLPFC). The integral value (IV) of blood oxygen signal was calculated as an indicator of cortical activation level and the whole brain correlation analysis was used to calculate cortical functional connectivity. The results showed that as motor fatigue deepened, the activation levels of bilateral DLPFC and PMC in early stage were significantly higher than those in later stage (P<0.05), and the functional connectivity strength of the motor related cortex areas between the hemispheres was significantly reduced, which was manifested by the functional connectivity strength of LSMC-RSMC and LPMC-RSMC showed a significant decrease in middle stage compared with that in early stage (P<0.05) and that the functional connectivity strength of LPMC-RSMC and RSMC-SMA showed a significant decrease in later stage compared with that in early stage (P<0.05). In each stage, the motor related cortex areas maintained high activation levels and the cerebral cortex showed extensive functional connectivity.Clinical Relevance- The clinical relevance of this study is to deepen the understanding of the neural processes related to upper limb resistance training based on motor fatigue, and provide a clinical basis for optimizing resistance training strategies related to motor dysfunction patients with altered brain function under fatigue.


Crystal structure of the dengue virus RNA-dependent RNA polymerase catalytic domain at 1.85-angstrom resolution.
Dengue fever, a neglected emerging disease for which no vaccine or antiviral agents exist at present, is caused by dengue virus, a member of the Flavivirus genus, which includes several important human pathogens, such as yellow fever and West Nile viruses. The NS5 protein from dengue virus is bifunctional and contains 900 amino acids. The S-adenosyl methionine transferase activity resides within its N-terminal domain, and residues 270 to 900 form the RNA-dependent RNA polymerase (RdRp) catalytic domain. Viral replication begins with the synthesis of minus-strand RNA from the dengue virus positive-strand RNA genome, which is subsequently used as a template for synthesizing additional plus-strand RNA genomes. This essential function for the production of new viral particles is catalyzed by the NS5 RdRp. Here we present a high-throughput in vitro assay partly recapitulating this activity and the crystallographic structure of an enzymatically active fragment of the dengue virus RdRp refined at 1.85-A resolution. The NS5 nuclear localization sequences, previously thought to fold into a separate domain, form an integral part of the polymerase subdomains. The structure also reveals the presence of two zinc ion binding motifs. In the absence of a template strand, a chain-terminating nucleoside analogue binds to the priming loop site. These results should inform and accelerate the structure-based design of antiviral compounds against dengue virus.


South American National Contributions to Knowledge of the Effects of Endocrine Disrupting Chemicals in Wild Animals: Current and Future Directions.
Human pressure due to industrial and agricultural development has resulted in a biodiversity crisis. Environmental pollution is one of its drivers, including contamination of wildlife by chemicals emitted into the air, soil, and water. Chemicals released into the environment, even at low concentrations, may pose a negative effect on organisms. These chemicals might modify the synthesis, metabolism, and mode of action of hormones. This can lead to failures in reproduction, growth, and development of organisms potentially impacting their fitness. In this review, we focused on assessing the current knowledge on concentrations and possible effects of endocrine disruptor chemicals (metals, persistent organic pollutants, and others) in studies performed in South America, with findings at reproductive and thyroid levels. Our literature search revealed that most studies have focused on measuring the concentrations of compounds that act as endocrine disruptors in animals at the systemic level. However, few studies have evaluated the effects at a reproductive level, while information at thyroid disorders is scarce. Most studies have been conducted in fish by researchers from Brazil, Argentina, Chile, and Colombia. Comparison of results across studies is difficult due to the lack of standardization of units in the reported data. Future studies should prioritize research on emergent contaminants, evaluate effects on native species and the use of current available methods such as the OMICs. Additionally, there is a primary focus on organisms related to aquatic environments, and those inhabiting terrestrial environments are scarce or nonexistent. Finally, we highlight a lack of funding at a national level in the reviewed topic that may influence the observed low scientific productivity in several countries, which is often negatively associated with their percentage of protected areas.


Pandemic influenza A (H1N1) 2009 in the English- and Dutch-speaking Caribbean: an epidemiological overview.
To describe epidemiological trends of pandemic influenza A (H1N1) in the English and Dutch-speaking Caribbean during the pandemic period. Data on laboratory-confirmed cases and deaths associated with pandemic influenza A (H1N1) contained in two regional databases at the Caribbean Epidemiology Centre (CAREC) were analysed. The data sources were epidemiological and laboratory reports from English and Dutch-speaking countries and the CAREC laboratory information system (LABIS). In the English- and Dutch-speaking Caribbean, pandemic influenza A (H1N1) was the predominant circulating influenza virus type during the pandemic period. There were three distinct phases: a first pandemic wave during mid-April to end of August 2009 (734 cases), a second pandemic wave during September-December 2009 (570 cases) and a phase of low transmission during January to mid-August 2010 (55 cases). The majority of cases (76%) were aged less than 30 years, with children of school age being most affected. Most cases (89%) presented with symptoms of the respiratory tract and smaller proportions (20-40%) presented with gastrointestinal and other symptoms. No cases tested were resistant to oseltamivir. A quarter of cases required hospitalization and the case fatality rate was 1.8%. The epidemiological characteristics of the pandemic in the English- and Dutch-speaking Caribbean were consistent with that in other parts of the world. It is important that post pandemic surveillance (epidemiological and virological) for respiratory illnesses continues to be enhanced in order to give a better understanding of seasonality and changing trends in respiratory illnesses and their aetiologic agents.


Sudden sensorineural hearing loss: subclinical viral and toxoplasmosis infections as aetiology and how they alter the clinical course.
To explore in a prospective study the evidence of certain viral and toxoplasmosis infections in sudden sensorineural hearing loss (SSHL). 84 consecutive patients with SSHL meeting certain criteria. All patients were assessed for specific IgM antibodies against cytomegalovirus, herpes simplex virus, toxoplasma and Epstein-Barr virus. All were treated with intravenous steroids and assigned to two groups: 76 IgM negative (NV group) and 8 IgM positive (no history of acute infection - V group). The mean hearing level at presentation was 86.5 dB HL (median, 100) in the V group and 60.7 dB HL (median, 61) in the NV group. The difference was statistically significant (p = 0.003). The mean hearing level following treatment was 81.8 dB HL (median, 88) in the V group and 48.7 dB HL (median, 39) in the NV group. The difference was statistically significant (p = 0.004). There was a considerable improvement in hearing after treatment only in the NV group (p < 0.000001). Recent subclinical viral or toxoplasmosis infections may be involved in the pathogenesis of SSHL (in approx. 10% of cases), suggesting that SSHL is not a single disease. When certain viruses or toxoplasmoses are involved, the hearing is much worse in comparison to patients with no such indication of infection. An alteration in treatment dosage or method of steroid administration may be needed in such cases.


NF kappa B-inducing kinase deficiency results in the development of a subset of regulatory T cells, which shows a hyperproliferative activity upon glucocorticoid-induced TNF receptor family-related gene stimulation.
CD4+CD25+ regulatory T cells (T(reg)) play an important role in maintaining immunologic tolerance. Glucocorticoid-induced TNFR family-related gene (GITR) expressed preferentially at high levels on T(reg) has been shown to be a key player of regulating T(reg)-mediated suppression. A recent study reports that NF-kappaB-inducing kinase (NIK) expression in thymic stroma is important for the normal production of T(reg) but not for its suppression capacity. In this report, we have shown that T(reg) from NIK-deficient mice display hyperproliferative activities upon GITR stimulation through an IL-2-independent mechanism. Furthermore, high dose IL-2, anti-CD28 stimulation, or GITR ligand-transduced bone marrow-derived dendritic cells used as APC (culture conditions which drive T(reg) proliferation in vitro) could not ablate this difference in proliferative activity between NIK-deficient and wild-type T(reg). Additional experiments have shown NIK-deficient mice have a higher ratio of CD4+CD25+CD62L(low) T(reg) both in thymus and periphery than their wild-type littermates. This CD62(low) subset is responsible for the hyperproliferative activity upon GITR stimulation. These data suggest a novel role of NIK in controlling the development and expansion of CD4+CD25+ regulatory T cells.


Atomic frequency reference at 1033 nm for ytterbium (Yb)-doped fiber lasers and applications exploiting a rubidium (Rb) 5S1/2 to 4D5/2 one-colour two-photon transition.
We demonstrate a two-photon transition of rubidium (Rb) atoms from the ground state (5S1/2) to the excited state (4D5/2), using a home-built ytterbium (Yb)-doped fiber amplifier at 1033 nm. This is the first demonstration of an atomic frequency reference at 1033 nm as well as of a one-colour two-photon transition for the above energy levels. A simple optical setup is presented for the two-photon transition fluorescence spectroscopy, which is useful for frequency stabilization for a broad class of lasers. This spectroscopy has potential applications in the fiber laser industry as a frequency reference, particularly for the Yb-doped fiber lasers. This two-photon transition also has applications in atomic physics as a background- free high- resolution atom detection and for quantum communication, which is outlined in this article.


Autosomal trisomy 18 and 13 syndromes in Ibadan, Nigeria.
The clinical, cytogenetic, dermatoglyphic and autopsy findings in 14 cases of trisomy 18 as well as in seven cases of trisomy 13 seen in the Paediatric Department of University College Hospital (UCH), Ibadan, Nigeria, from 1972 to 1981 are presented. The incidence was 0.12 and 0.08 per 1000 livebirths for trisomies 18 and 13 respectively. All the patients were of regular trisomies except two; one case was mosaic for trisomy 18 and another case of trisomy 13 seemed to have derived an extra chromosome of the D group from a reciprocal D/D translocation that was maternally carried. Both syndromes have certain physical features in common which sometimes made clinical diagnosis difficult, but each had diagnostic features of its own. While the combination of microcephaly, cleft lip and/or palate, microphthalmos, capillary haemangioma and polydactyly form the hallmark of trisomy 13, the presence of prominent occipit, micrognathia, clenched fist together with protruding heel make the appearance of trisomy 18 unmistakable. The prognosis in both syndromes is poor and those who survive beyond infancy exhibit severe mental retardation. It is therefore important to establish the diagnosis early so as to conserve limited resources in an attempt to prolong the lives of neonates with these syndromes.


Salbutamol: beware of the paradox!
A 7-year-old known asthmatic presented with an acute severe asthma attack to the Accident and Emergency department. Following a poor response to salbutamol and ipratropium nebulisers, he was given intravenous salbutamol and aminophylline. Over the course of the following 3 h, there was improvement in his bronchospasm with decreasing oxygen requirement, however, his respiratory rate showed an upward trend. Serial blood gas estimations showed a worsening metabolic acidosis unresponsive to two fluid boluses of 20 ml/kg of normal saline. Lactate levels were subsequently measured and found to be high, accounting for the metabolic acidosis. High lactate levels were attributed to intravenous salbutamol. His blood gases and lactate level returned to normal within 3 h after stopping intravenous salbutamol. He was recommended on salbutamol nebulisers while still continuing on intravenous aminophylline. He continued to improve and was discharged home after 4 days.


The existence and characteristics of rats and shrews in endemic leptospirosis areas and types of ectoparasites: a case study of West Jakarta, Indonesia.
Background: This study aimed to determine the presence and species of the rats and shrews that can potentially cause leptospirosis in West Jakarta, Indonesia, and the species of ectoparasites found in them. Methods: The research was a descriptive study employing a cross-sectional approach. The study population was all species of rats and shrews in the region and the sample collection technique used was purposive sampling. The traps were installed in the homes of respondents who had suffered from leptospirosis and their closest neighbors, with a total of 521 traps. Leptospirosis data based on secondary data was obtained from West Jakarta Health Office (2016-August 2019). The technique for catching rats involved using humane live traps, while the identification of the rats and ectoparasites was done in the laboratory. Results: It was found that more rats were caught in Cengkareng Timur sub-district, Cengkareng District, with a percentage of 14.8%, while the least in Duri Kepa, Kapuk, Kedaung Kali Angke and Kedoya Utara with a percentage of 3.7%. The rats were mostly found in East Cengkareng Sub-District, with the most common type being Rattus rattus (74.1 %) and the least Suncus murinus (11.1%); more male rats were caught (66.7%) than female (33.3%). The type of ectoparasite found in the rats was fleas. Xenopsylla cheopis was the most common type, at 83.3% and more fleas were male, at 66.7%. The most common rat species was Rattus rattus. The ectoparasite most commonly found in them was the female flea Xenopsylla cheopis.Conclusions: Rattus rattus and Xenopsylla cheopis were found in an East Cengkareng sub-district. Surveys, monitoring, and control of rats and ectoparasites are essential for the preparedness and development of an early warning system of possible diseases that they can cause.


[Altering of phragmoplast orientation as a result of excessive centrifugal movement in the absence of cell plate].
The inability of phragmoplast to stop its centrifugal movement after reaching the mother cell membrane is described in abnormal meiosis with the arrest of cell plate formation. The excess of phragmoplast expansion leads to rotation of the whole telophase figure (phragmoplast with daughter nuclei) within the cell through 90 degrees. It has been suggested that this phenomenon may occur because of a the lack of signal stopping cytokinesis. Such a signal arises due to formation of daughter cell membranes.


Glycosylated hemoglobin concentration for assessment of glycemic control in diabetic cats.
Blood glycosylated hemoglobin (GHb) concentration was quantified in 84 healthy cats, 9 cats with stress-induced hyperglycemia, 37 cats with newly diagnosed diabetes mellitus, and 122 diabetic cats treated with insulin or glipizide. Diabetic control was classified as good or poor in insulin-treated or glipizide-treated cats based on review of history, physical examination findings, changes in body weight, and measurement of blood glucose concentrations. Blood GHb concentration was determined using an affinity chromatography assay. Mean blood GHb concentration was similar for healthy normoglycemic cats and cats with transient, stress-induced hyperglycemia, but was significantly (P < .001) higher in untreated diabetic cats when compared with healthy normoglycemic cats. Mean blood GHb concentration was significantly (P < .001) higher in 84 cats with poorly controlled diabetes mellitus when compared with 38 cats in which the disease was well controlled. Mean blood GHb concentration decreased significantly (P < .01) in 6 cats with untreated diabetes mellitus after insulin and dietary treatment. A similar significant (P < .01) decrease in mean blood GHb concentration occurred in 7 cats with poorly controlled diabetes mellitus after diabetic control was improved by an increase in insulin dosage from 1.1 +/- 0.9 to 1.4 +/- 0.6 U/kg/ 24 h and by feeding a diet containing increased fiber content and in 6 cats with transient diabetes mellitus 8.2 +/- 0.6 weeks after discontinuing insulin treatment. There was a significant (P < .01) stress-induced increase in mean fasting blood glucose concentration and mean blood glucose concentration for 12 hours after administration of insulin or glipizide but no change in mean blood GHb concentration in 5 docile diabetic cats 12.2 +/- 0.4 weeks after the cats became fractious as a result of frequent hospitalizations and blood samplings. Results of this study suggest that evaluation of blood GHb concentration may be a clinically useful tool for monitoring glycemic control of diabetes in cats.


Human periodontal ligament fibroblasts synthesize C-reactive protein and Th-related cytokines in response to interleukin (IL)-6 trans-signalling.
To characterize the potential of human periodontal ligament fibroblasts (HPLF) to synthesize CRP and Th-related cytokines in response to IL-6 in periodontal health and apical inflammation. Primary HPLF stimulated with IL-6, soluble(s) IL-6 receptor (R) and controls were assayed for CRP, Th1, Th2, Th17 and Treg-related cytokines by quantitative real-time PCR and ELISA, respectively. IL-6R mRNA expression and its soluble protein levels were screened in HPLF cultures, and ex vivo samples of healthy periodontal ligaments (n = 5) and apical lesions (n = 13). Data were analysed with ANOVA or unpaired t-test. 0.5 ng mL-1 IL-6 plus 1 ng mL-1 of its soluble receptor (sIL-6R) for 24 h was effective in inducing CRP production. IL-6 alone had a mild dose-dependent effect; co-stimulation with sIL-6R significantly enhanced this effect, whereas it was completely abolished by the addition of IL-6R blocking antibody (P < 0.05). Similarly, higher mRNA expression and protein levels of Th1, Th17 and partially Treg-related cytokines were found for IL-6 combined with its soluble receptor versus the nonstimulated group and IL-6R antibody (P < 0.05). IL-6R mRNA expression was slightly induced by IL-6 compared to THP-1 cells, but sILR-6 protein could not be detected in HPLF. High sIL-6R levels were detected in apical lesions and were immunolocalized to mononuclear inflammatory cells and proliferating epithelium. IL-6 trans-signalling induced Th1 and Th17-related cytokines and represents an extra-hepatic mechanism for PCR synthesis in human periodontal ligament fibroblasts, contributing to explain the bone-destructive phenotype of apical lesions and eventually its systemic complications.


Interference competition as a key determinant for spatial distribution of mangrove crabs.
The spatial distribution of mangrove crabs has been commonly associated with tree zonation and abiotic factors such as ground temperature and soil granulometry. Conversely, no studies were designed to investigate the role of competition for resources and predation in shaping crab distribution in mangroves, despite these biotic factors are recognised as key determinants for spatial patterns observed in the communities colonising rocky and sandy intertidal habitats.We studied floral and faunal assemblages in two zones of a Sri Lankan mangrove, a man-made upper intertidal level and a natural eulittoral, mid-shore one. Leaf choice experiments were designed to study both feeding rate and intra and inter-specific interactions for food of sesarmid crabs in the two habitats in order to better understand crab spatial distribution. The two intertidal belts differed in terms of floral composition and crab species abundance. The eulittoral zone was strongly dominated by Neosarmatium smithi, while within the elevated littoral fringe four sesarmids (N. smithi, N. asiaticum, N. malabaricum and Muradium tetragonum) were more evenly distributed. At both levels, all sesarmids showed to collect significantly more Bruguiera spp. and Rhizophora apiculata leaves than Excoecaria agallocha ones. There was no temporal segregation in feeding activity among the four species, resulting in a high interference competition for leaves. Regardless of the habitat, N. smithi was always successful in winning inter-specific fights. Our results showed that the elevated littoral fringe was more crowded with crabs, but was less favourable in terms of food availability and environmental conditions. The dominance of N. smithi in gathering mangrove leaves suggests that this species may segregate the other sesarmids into less favourable habitats. The present data strongly suggest for the first time that interference competition for food can contribute to shape mangrove crab spatial distribution.


Vasovasostomy and predictors of vasal patency: a systematic review.
About 3-6% of vasectomized men requested vasectomy reversal, for various reasons. Vasal patency (VP) is an important surrogate outcome of vasectomy reversal. This article reviews the impact of surgical skills, surgical approaches, intraoperative vasal fluid characteristics and the length of obstructive interval on VP. Based on the best available evidence, the rate of patency is related to the operative frequency of the surgeons, with better results obtained by surgeons who perform the operations at least 10 times annually. Microsurgical vasovasostomy is the preferred technique for durable good results. One-layer vasovasostomy and two-layer vasovasostomy seem to be equal with regard to VP. The rate of patency following vasovasostomy in the convoluted vas and vasovasostomy in the straight vas is comparable. The patency rate is high in men with clear intraoperative vasal fluid in at least one vas. VP is still high among patients with a long obstructive interval. In conclusion, surgical skills and intraoperative vasal fluid characteristics are the most important predictors of VP. Postoperative semen quality and the age of the female partner determine the chance of spontaneous conception in these couples.


[Robotic-assisted organ transplantation].
Advanced surgical procedures have traditionally been a domain of open surgery. However, minimally invasive approaches are evolving with the development of robotic technology which appears capable to overcome technical limitations of conventional laparoscopy. While traditionally perceived as impossible indications for minimally invasive surgery, reports on robotic organ transplantations have surfaced with promising results.


Effects of dosage titration of methylprednisolone acetate and triamcinolone acetonide on interleukin-1-conditioned equine articular cartilage explants in vitro.
Osteoarthritis is a frequent sequela of joint disease, especially with severe injuries or if attempts at therapy are unsuccessful. Negative and positive effects of corticosteroid treatment of articular cartilage have been demonstrated by in vitro and in vivo studies. To assess the metabolic effects of varying dosages of methylprednisolone acetate (MPA) and triamcinolone acetonide (TA) on interleukin-1alpha (IL-1) conditioned equine cartilage explants. Our hypothesis was that lower dosages of corticosteroids would be less detrimental to cartilage metabolism than higher dosages. TA would be less detrimental to cartilage metabolism than MPA. Treatment groups included articular cartilage explants with no IL-1 (control), IL-1 alone, and IL-1 plus 10, 5, 1 and 0.5 mg/ml MPA or 1.2, 0.6, 0.12 and 0.06 mg/ml TA. Explants were labelled with 35SO4 prior to the beginning and end of the experiment to assess glycosaminoglycan (GAG) degradation and synthesis, respectively. Total GAG content in media and explants and total cartilage DNA were also analysed. MPA and TA reduced GAG synthesis compared to control and IL-1 alone. The highest dosage of MPA (10 mg/ml) reduced GAG synthesis less than lower dosages of MPA and all dosages of TA. Compared to IL-1 alone, all dosages of TA and lower dosages of MPA increased GAG degradation. MPA at 10 mg/ml reduced GAG degradation. Both MPA and TA increased media GAG content compared to control and IL-1 explants. Total cartilage GAGs were unchanged with MPA, but reduced with TA, compared with IL-1 alone. Total cartilage DNA was decreased with MPA and increased with TA compared to IL-1 and control explants. MPA and TA did not counteract the negative effects of IL-1 and did not maintain cartilage metabolism at control levels. Lower dosages of MPA and TA were not less detrimental to cartilage metabolism than higher dosages. TA did not appear to be less harmful than MPA on cartilage metabolism. The results of this study differ from the findings of comparable in vivo studies. The low numbers of horses used in this study limits extrapolation of these findings to the equine population; however, this study also questions the clinical relevance of this in vitro model.


Hereditary leiomyomatosis and renal cell cancer in families referred for fumarate hydratase germline mutation analysis.
Heterozygous fumarate hydratase (FH) germline mutations cause hereditary leiomyomatosis and renal cell cancer (HLRCC), an autosomal dominant syndrome characterized by multiple cutaneous piloleiomyomas, uterine leiomyomas and papillary type 2 renal cancer. The main objective of our study was to evaluate clinical and genetic data from families suspected of HLRCC on a nationwide level. All families referred for FH mutation analysis in the Netherlands were assessed. We performed FH sequence analysis and multiplex ligation-dependent probe amplification. Families with similar FH mutations were examined for haplotype sharing. In 14 out of 33 families, we identified 11 different pathogenic FH germline mutations, including 4 novel mutations and 1 whole-gene deletion. Clinical data were available for 35 FH mutation carriers. Cutaneous leiomyomas were present in all FH mutation carriers older than 40 years of age. Eleven out of 21 female FH mutation carriers underwent surgical treatment for symptomatic uterine leiomyomas at an average of 35 years. Two FH mutation carriers had papillary type 2 renal cancer and Wilms' tumour, respectively. We evaluated the relevance of our findings for clinical practice and have proposed clinical diagnostic criteria, indications for FH mutation analysis and recommendations for management.


Nutritional characteristics of middle-aged Japanese vegetarians.
Despite the possible overall health benefits of a vegetarian diet, research about the nutritional characteristics of Japanese vegetarians is small. Our objective was to investigate the nutritional characteristics of Japanese vegetarians compared with Japanese non-vegetarians. The dietary intake, anthropometric and biochemical status of 75 middle-aged Japanese vegetarians (JV, 20 men and 55 women) were compared with those of 50 age- and sex- matched middle-aged Japanese non-vegetarians (JNV, 32 men and 18 women) in a cross-sectional study. JV men had significantly higher calcium, iron (p<0.001) and dietary fiber (p<0.01), and significantly lower vitamin B(12), cholesterol, animal fat intake and percentage of energy as animal protein (p<0.01) than JNV men. In addition, JV men had significantly lower body mass index (p<0.05), diastolic blood pressure (p<0.001), systolic blood pressure (p<0.01), aspartate transaminase, alanin transaminase (p<0.05) and serum triacylglyceride (p<0.001) than JNV men. JV women had significantly lower systolic pressure and serum triacylglyceride (p<0.05) than JNV women. JV men and women had better nutritional characteristics than JNV men and women from the standpoint of lifestyle-related diseases.


A Scalable, Solution-Based Approach to Tuning the Solubility and Improving the Photoluminescence of Chemically Exfoliated MoS2.
MoS2 are two-dimensional (2D) materials that exhibit emerging photoluminescence (PL) at the monolayer level and have potential optoelectronic applications. Monolayers of MoS2 typically achieved by mechanical exfoliation (Me), chemical vapor deposition (CVD), and chemical exfoliation (Ce) via lithium intercalation contain numerous defects that significantly reduce their PL efficiency. Several studies have reported overcoming poor PL in mechanically exfoliated and CVD-grown MoS2, but such studies for chemically exfoliated MoS2 (Ce-MoS2) have not been reported. Here, we report a solution-based method of enhancing the PL of Ce-MoS2 by reacting with molecules with suitable functional groups at high temperatures. Reaction with dodecanethiol (DDT) generates PL that is more intense than mechanically exfoliated MoS2 (Me-MoS2) with high crystallinity and has a significantly broader range of wavelengths. Based on ultraviolet-visible, Fourier transform infrared, X-ray photoemission, and PL spectroscopy as well as transmission electron and PL imaging, we propose that the present method modifies PL properties of Ce-MoS2 by simultaneously annealing, replacing molybdenum-oxygen with molybdenum-sulfur bonds, inducing strain, and generating a nanopolycrystalline structure. This work points to such defect engineering using molecules as an effective means to modify the properties of Ce-MoS2 and layered transition-metal dichalcogenides more generally.


Extracellular vesicles from bone marrow-derived mesenchymal stem cells protect against murine hepatic ischemia/reperfusion injury.
Hepatic ischemia/reperfusion injury (IRI) and associated inflammation contributes to liver dysfunction and complications after liver surgery and transplantation. Mesenchymal stem cells (MSCs) have been reported to reduce hepatic IRI because of their reparative immunomodulatory effects in injured tissues. Recent studies have highlighted beneficial effects of extracellular vesicles from mesenchymal stem cells (MSC-EV) on tissue injury. The effects of systemically administered mouse bone marrow-derived MSC-EV were evaluated in an experimental murine model of hepatic IRI induced by cross-clamping the hepatic artery and portal vein for 90 minutes followed by reperfusion for periods of up to 6 hours. Compared with controls, intravenous administration of MSC-EV 30 minutes prior to IRI dramatically reduced the extent of tissue necrosis, decreased caspase 3-positive and apoptotic cells, and reduced serum aminotransferase levels. MSC-EV increased hepatic messenger RNA (mRNA) expression of NACHT, LRR, and PYD domains-containing protein 12, and the chemokine (C-X-C motif) ligand 1, and reduced mRNA expression of several inflammatory cytokines such as interleukin 6 during IRI. MSC-EV increased cell viability and suppressed both oxidative injury and nuclear factor kappa B activity in murine hepatocytes in vitro. In conclusion, the administration of extracellular vesicles derived from bone marrow-derived MSCs may ameliorate hepatic IRI by reducing hepatic injury through modulation of the inflammatory response.Liver Transplantation 23 791-803 2017 AASLD.


Dietary risk factors for rheumatic diseases.
Recent scientific data illuminate the dietary link to rheumatic disorders. This review summarizes recently published articles on the dietary link to rheumatoid arthritis, gout, and osteoarthritis. A prospective study suggests that higher intakes of meat and total protein as well as lower intakes of fruit, vegetables, and vitamin C are associated with an increased risk of inflammatory polyarthritis or rheumatoid arthritis. Several studies suggest that the Mediterranean-type diet or its main components may have protective effects on the development or severity of rheumatoid arthritis. A recent prospective study investigated several purported dietary factors for gout and confirmed some of the long-standing suspicions (red meats, seafood, beer, and liquor), exonerated others (total protein, wine, and purine-rich vegetables), and also identified potentially new protective factors (dairy products). Recent double-blind, randomized, placebo-controlled studies suggest that antioxidant vitamins (vitamin E, vitamin C, beta-carotene, and retinol) do not halt the progression of symptomatic knee osteoarthritis, as was previously suggested. Because diet is an unavoidable universal exposure for people, even a small effect that can be achieved by dietary manipulation may produce a large impact on the population's health. As the evidence on the role of dietary factors in rheumatic disorders grows it becomes increasingly important for clinicians and investigators in the field of rheumatology to familiarize themselves with the relevant data and appropriately apply them to clinical and public health practice.


How many or how much? Testing the relative influence of the number of social network risks versus the amount of time exposed to social network risks on post-treatment substance use.
Having high-risk, substance-using friends is associated with young adult substance use disorder (SUD) relapse. It is unclear, however, whether it is the total number of high-risk friends, or the amount of time spent with high-risk friends that leads to relapse. Unclear also, is to what extent low-risk friends buffer risk. This study examined the influence of number of high-risk and low-risk friends, and the amount time spent with these friends on post-treatment percent days abstinent (PDA). Young adult inpatients (N=302) were assessed at intake, and 3, 6, and 12 months on social network measures and PDA. Mixed models tested for effects of number of high- and low-risk friends, and time spent with these friends on PDA, and for net-risk friend effects to test whether low-risk friends offset risk. Within and across assessments, number of, and time spent with high-risk friends was negatively associated with PDA, while the inverse was true for low-risk friends. Early post-treatment, time spent with friends more strongly predicted PDA than number of friends. Participants were more deleteriously affected by time with high-risk friends the longer they were out of treatment, while contemporaneously protection conferred by low-risk friends increased. This interaction effect, however, was not observed with number of high- or low-risk friends, or number of friends net-risk. Young adult SUD patients struggling to break ties with high-risk friends should be encouraged to minimize time with them. Clinicians should also encourage patients to grow their social network of low-risk friends.


Polysynaptic neuronal pathways from tail cutaneous afferents to hindlimb motoneurons in the spinalized cat.
Postsynaptic potentials (PSPs) were recorded in motoneurons innervating the m. posterior biceps and semitendinosus (PBSt-MN) and m. triceps surae (GS-MN) in 19 spinalized adult cats, after electrical stimulation of the dorsal and ventral tail cutaneous nerves (TDC and TVC). With stimulation at 1,5-5 time threshold, inhibitory PSPs (IPSPs), excitatory PSPs (EPSPs), and mixed PSPs (IPSP/EPSPs, EPSP/IPSPs) were recorded in approximately 90% of PBSt-MN and 70% of GS-MN. IPSPs (IPSPs, IPSP/EPSPs) after stimulation of the contralateral TVC and TDC were observed in 54% and 52% of PBSt motoneurons, respectively. EPSPs (EPSP, EPSP/IPSPs) were recorded after stimulation of ipsilateral TVC in 56% of PBSt motoneurons. IPSPs produced after stimulation of ipsilateral TDC and TVC were observed in 65% and 63%, respectively, of GS-MN showing PSPs after stimulation of tail cutaneous nerves. Measurement of segmental latencies of the earliest PSPs (PBSt-MN, IPSPs: 2.1-7.4 ms, EPSPs: 3.1-18.9 ms, GS-MN, IPSPs: 2.5-17.1 ms, EPSPs: 4.2-19.4 ms) suggested that most of the neural pathways from tail cutaneous afferents to hindlimb motoneurons are at least trisynaptic in the L7 spinal segments. Hemisection of spinal cord at S1-S2 indicated that neural pathways from both the ipsilateral and contralateral low threshold cutaneous afferents pass through the ipsilateral spinal cord at S1-S2. The neural pathways from tail cutaneous afferents to hindlimb motoneurons to maintain the balance of the pelvic girdle were discussed.


Pathologic effects in brain after intracranial pressure monitoring in clinically normal dogs, using a fiberoptic monitoring system.
During 2 separate studies, intracranial pressure (ICP) was measured in 13 healthy dogs (group A, n = 7; group B, n = 6), using a fiberoptic monitoring system implanted surgically in the right superficial cerebral cortex. Average ICP was measured for 15 minutes after a 15-minute postimplantation period of equilibration. Intracranial pressure was measured in group-A dogs at 2.0 and 1.3% end-tidal isoflurane concentrations. Mean +/- 1 SD ICP in group-A dogs at 2.0 and 1.3% end-tidal isoflurane concentrations was 11 +/- 2 and 11 +/- 3 mm of Hg, respectively. Dogs of group A were euthanatized immediately after measurements were obtained. Mean ICP +/- 1 SD in group-B dogs was 11 +/- 3 mm of Hg. After monitoring, but prior to euthanasia, group-B dogs underwent callosotomy, and were maintained for 30 days after surgery. The brain was removed from all dogs, formalin fixed, and examined grossly and microscopically for lesions associated with fiberoptic cable implantation. Variable degrees of hemorrhage and mechanical brain damage were seen focally around the catheter site in all brains from group-A dogs, especially when the cable entered through a sulcus. In 1 dog, local vacuolation was seen in the brain immediately adjacent to the tract associated with implantation of the fiberoptic catheter. In all other dogs, the additional cortex was histologically normal. Histologic lesions associated with cable implantation were not observed in group-B dogs.


Incidence of fatigue symptoms and diagnoses presenting in UK primary care from 1990 to 2001.
Little is known about whether the incidence of symptoms of fatigue presented in primary care, and the consequent diagnoses made, change over time. The UK General Practice Research Database was used to investigate the annual incidence of both fatigue symptoms and diagnoses recorded in UK primary care from 1990 to 2001. The overall incidence of all fatigue diagnoses decreased from 87 per 100 000 patients in 1990 to 49 in 2001, a reduction of 44%, while postviral fatigue syndromes decreased from 81% of all fatigue diagnoses in 1990 to 60% in 2001. Chronic fatigue syndrome (CFS) and myalgic encephalomyelitis (ME) together increased from 9% to 26% of all fatigue diagnoses. The incidence of fibromyalgia increased from less than 1 per 100 000 to 35 per 100 000. In contrast, there was no consistent change in the incidence of all recorded symptoms of fatigue, with an average of 1503 per 100 000, equivalent to 1.5% per year. CFS/ME and fibromyalgia were rarely diagnosed in children and were uncommon in the elderly. All symptoms and diagnoses were more common in females than in males. The overall incidence of fatigue diagnoses in general has fallen, but the incidence rates of the specific diagnoses of CFS/ME and fibromyalgia have risen, against a background of little change in symptom reporting. This is likely to reflect fashions in diagnostic labelling rather than true changes in incidence.


Nasal infection of Alcaligenes bronchisepticus (Bordetella bronchiseptica) and lesions in newborn rabbits.
The mode of experimental infection with Alcaligenes bronchisepticus (Bordetella bronchiseptica) of pig and rabbit origin, and lesions in the respiratory organs were examined in three groups, A, B and C, of newborn rabbits. Groups A and C were free from the organism and agglutinating antibody and inoculated with the organisms of pig and rabbit origin, respectively. Group B had maternal antibody and was inoculated with the organism of pig origin. Establishment and persistence of infection with the organism were certified in the nasal cavities, trachea, or lungs of groups A and C 3 days after inoculation or later. Late establishment of infection occurred in trachea and lungs of group B. Agglutinating antibody was detected in groups A and B mostly 14 days after inoculation or later. Ventral turbinate atrophy occurred in groups A and B mostly 10 days after inoculation or later. Histologically, it was hypo-osteogenesis caused by degeneration of osteoblasts and proliferation of fibroblast-like cells in the osseous tissue. Catarrhal inflammation in the nasal and tracheal mucous membranes, and bronchopneumonia with peribronchiolitis developed commonly in all the groups. The fluorescent antibody technique revealed antigen of the organism of pig origin on the nasal mucosa, mostly of the dorsal and ventral meatus, and on the tracheal and bronchiolar mucosa in groups A and B.


Coronary heart disease mortality in relation to dietary, lifestyle and biochemical risk factors in the countries of the Seven Countries Study: a secondary dataset analysis.
The present study explored coronary heart disease (CHD) mortality rates in 2011 in countries that participated in the Seven Countries Study (SCS) in relation to several dietary and anthropometric/biochemical risk factors. Special focus was given to Crete and the Ionian Islands. This was a cross-sectional study of secondary analysis of databases using data from the World Health Organization, the Food and Agriculture Organization database and the Greek National Cadaster and Cartography Organization. Geographically weighted regression was applied to identify the high-risk regions in relation to the significant factors. Crete, the Ionian Islands and Japan had the lowest mortality rates (28.9, 30.1 and 31.2 deaths/100,000 people, respectively) in contrast to Serbia/Montenegro that presented the highest rates (105.1 deaths/100,000 people). Diet, physical inactivity and hypertension were found to be the most significant factors in the model (P < 0.05). Regions of no risk were Crete, Ionian Islands and Japan (exponent B = 0.65; 95% confidence interval = 0.293-0.942; P < 0.001), whereas Serbia/Montenegro and Finland were identified as high-risk areas with a 2.97-fold higher probability for CHD mortality (95% confidence interval = 1.736-4.028; P < 0.001). Observed patterns of CHD mortality and related factors may be helpful for appropriate management by health planners when aiming to reduce its prevalence, particularly in high-risk areas.


Drosophila acetylcholinesterase: effect of post-translational [correction of post-traductional] modifications on the production in the baculovirus system and substrate metabolization.
Acetylcholinesterase cDNAs from Drosophila melanogaster modified on its primary sequence were cloned into baculovirus and were expressed in Sf9 cells with the aim to identify a mutant form that produces the enzyme at a high level. Directed mutagenesis was used in order to independently knockout different sites of post-translational modifications: exchange of the C-terminal hydrophobic peptide for a glycolipid molecule, dimerization by disulfide bridge, N-linked glycosylation at the five accessible sites, and subunit formation by proteolytic cleavage of a hydrophilic peptide found in the precursor. Another mutation involved the elimination of a free cysteine in the mature protein. All mutations involving post-translational modifications resulted in lower recoveries, suggesting that they are useful for maintaining high amounts of protein in the synapse. By contrast, elimination of a free cysteine in the mature protein permitted an increase in the level of production of the enzyme. These mutations did not affect specific activity of the enzyme at substrate concentrations ranging from 3 microM to 200 mM, suggesting that activation and inhibition of the enzyme activity does not originate from a polymorphism in post-translational modifications.


Hypertrophic Pachymeningitis in a Southern Chinese Population: A Retrospective Study.
Aims: To investigate the causes, clinical characteristics, imaging features, and therapeutic implications of hypertrophic pachymeningitis (HP) in a southern Chinese population. Methods: We retrospectively analyzed 48 patients with HP with different causes from 1 January 2006 to 31 December 2018. Clinical manifestation, laboratory findings, and neuroimaging results were evaluated in all HP patients. Results: The mean age at onset was 50 ± 12 years. The most common diagnosis was idiopathic HP (67%), followed by antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (15%), tuberculous meningitis (8%), viral meningitis (6%), and bacterial meningitis (4%). Headache was the most common symptom. The most frequently changed laboratory finding was elevated erythrocyte sedimentation rate (ESR). Imaging was characterized by cerebral or spinal dura mater enhancement in MRI scan with contrast. Enhancements were mainly located in the posterior fossa for idiopathic HP; frontal, parietal, and occipital lobes for ANCA-related HP; and posterior fossa for tuberculous-associated HP. Diffuse enhancement was found in most cases, except for tuberculous-associated HP. Glucocorticoid or immunosuppressive treatment was applied in most cases. Conclusions: The etiology of HP varied among patients, with idiopathic HP being the most common. MRI showed enhancement of the dura mater, which differed according to different etiologies. Glucocorticoid or immunosuppressive agents were the primary drugs for treatment.


A bright future for engineering piezoelectric 2D crystals.
The piezoelectric effect, mechanical-to-electrical and electrical-to-mechanical energy conversion, is highly beneficial for functional and responsive electronic devices. To fully exploit this property, miniaturization of piezoelectric materials is the subject of intense research. Indeed, select atomically thin 2D materials strongly exhibit the piezoelectric effect. The family of 2D crystals consists of over 7000 chemically distinct members that can be further manipulated in terms of strain, functionalization, elemental substitution (i.e. Janus 2D crystals), and defect engineering to induce a piezoelectric response. Additionally, most 2D crystals can stack with other similar or dissimilar 2D crystals to form a much greater number of complex 2D heterostructures whose properties are quite different to those of the individual constituents. The unprecedented flexibility in tailoring 2D crystal properties, coupled with their minimal thickness, make these emerging highly attractive for advanced piezoelectric applications that include pressure sensing, piezocatalysis, piezotronics, and energy harvesting. This review summarizes literature on piezoelectricity, particularly out-of-plane piezoelectricity, in the vast family of 2D materials as well as their heterostructures. It also describes methods to induce, enhance, and control the piezoelectric properties. The volume of data and role of machine learning in predicting piezoelectricity is discussed in detail, and a prospective outlook on the 2D piezoelectric field is provided.


miR-135a Inhibits Cancer Stem Cell-Driven Medulloblastoma Development by Directly Repressing Arhgef6 Expression.
microRNAs (miRNAs) are short noncoding RNAs, which regulate gene expression post-transcriptionally and play crucial roles in relevant biological and pathological processes. Here, we investigated the putative role of miRNAs in modulating the tumor-initiating potential of mouse medulloblastoma (MB)-derived cancer stem cells (CSCs). We first subjected bona fide highly tumorigenic (HT) CSCs as well as lowly tumorigenic MB CSCs and normal neural stem cells to miRNA profiling, which identified a HT CSC-specific miRNA signature. Next, by cross-checking CSC mRNA/miRNA profiles, we pinpointed miR-135a as a potential tumor suppressor gene, which was strongly downregulated in HT CSCs as well as in the highly malignant experimental tumors derived from them. Remarkably, enforced expression of miR-135a in HT CSCs strongly inhibited tumorigenesis by repressing the miR-135a direct target gene Arhgef6. Considering the upregulation of Arhgef6 in human MBs and its involvement in mediating experimental medulloblastomagenesis, its efficient suppression by miR-135a might make available an effective therapeutic strategy to selectively impair the tumorigenic potential of MB CSCs. Stem Cells 2015;33:1377-1389.


Fracture Risk and Association With TDF Use Among People With HIV in Large Integrated Health Systems.
Greater decline in bone health among people with HIV (PWH) has been documented but fracture risk and the impact of specific antiretroviral therapy (ART) regimens remain unclear. Retrospective analyses of electronic health record data from 3 US integrated health care systems. Fracture incidence was compared between PWH aged 40 years or older without prior fracture and demographically matched people without HIV (PWoH), stratified by age, sex, and race/ethnicity. Multivariable Cox proportional hazards models were used to estimate fracture risk associated with HIV infection. The association of tenofovir disoproxil fumarate (TDF) use and fracture risk was evaluated in a subset of PWH initiating ART. Incidence of fracture was higher in PWH [13.6/1000 person-years, 95% confidence interval (CI): 13.0 to 14.3, n = 24,308] compared with PWoH (9.5, 95% CI: 9.4 to 9.7, n = 247,313). Compared with PWoH, the adjusted hazard ratio (aHR) for fracture among PWH was 1.24 (95% CI: 1.18 to 1.31). The association between HIV infection and fracture risk increased with age, with the lowest aHR (1.17, 95% CI: 1.10 to 1.25) among those aged 40-49 years and the highest aHR (1.89, 95% CI: 1.30 to 2.76) among those aged 70 years or older. Among PWH initiating ART (n = 6504), TDF was not associated with significant increase in fracture risk compared with non-TDF regimens (aHR: 1.18, 95% CI: 0.89 to 1.58). Among people aged 40 years or older, HIV infection is associated with increased risk of fractures. Bone health screening from the age of 40 years may be beneficial for PWH. Large cohort studies with longer follow-up are needed to evaluate TDF effect and the potential benefit of early screening.


Discovery of New Zika Protease and Polymerase Inhibitors through the Open Science Collaboration Project OpenZika.
The Zika virus (ZIKV) is a neurotropic arbovirus considered a global threat to public health. Although there have been several efforts in drug discovery projects for ZIKV in recent years, there are still no antiviral drugs approved to date. Here, we describe the results of a global collaborative crowdsourced open science project, the OpenZika project, from IBM's World Community Grid (WCG), which integrates different computational and experimental strategies for advancing a drug candidate for ZIKV. Initially, molecular docking protocols were developed to identify potential inhibitors of ZIKV NS5 RNA-dependent RNA polymerase (NS5 RdRp), NS3 protease (NS2B-NS3pro), and NS3 helicase (NS3hel). Then, a machine learning (ML) model was built to distinguish active vs inactive compounds for the cytoprotective effect against ZIKV infection. We performed three independent target-based virtual screening campaigns (NS5 RdRp, NS2B-NS3pro, and NS3hel), followed by predictions by the ML model and other filters, and prioritized a total of 61 compounds for further testing in enzymatic and phenotypic assays. This yielded five non-nucleoside compounds which showed inhibitory activity against ZIKV NS5 RdRp in enzymatic assays (IC50 range from 0.61 to 17 μM). Two compounds thermally destabilized NS3hel and showed binding affinity in the micromolar range (Kd range from 9 to 35 μM). Moreover, the compounds LabMol-301 inhibited both NS5 RdRp and NS2B-NS3pro (IC50 of 0.8 and 7.4 μM, respectively) and LabMol-212 thermally destabilized the ZIKV NS3hel (Kd of 35 μM). Both also protected cells from death induced by ZIKV infection in in vitro cell-based assays. However, while eight compounds (including LabMol-301 and LabMol-212) showed a cytoprotective effect and prevented ZIKV-induced cell death, agreeing with our ML model for prediction of this cytoprotective effect, no compound showed a direct antiviral effect against ZIKV. Thus, the new scaffolds discovered here are promising hits for future structural optimization and for advancing the discovery of further drug candidates for ZIKV. Furthermore, this work has demonstrated the importance of the integration of computational and experimental approaches, as well as the potential of large-scale collaborative networks to advance drug discovery projects for neglected diseases and emerging viruses, despite the lack of available direct antiviral activity and cytoprotective effect data, that reflects on the assertiveness of the computational predictions. The importance of these efforts rests with the need to be prepared for future viral epidemic and pandemic outbreaks.


Desamino arginine vasopressin (DDAVP) as a diagnostic agent.
The use of 1-desamino-8-arginine vasopressin (DDAVP) was examined as a diagnostic agent, within the context of the comprehensive one day renal function test. Thirty-three normal young subjects were examined in an 8 1/2 hour day. It was found that a 20 microgram intranasal dose of DDAVP consistently resulted in the production of a urinary osmolality to 800mosmol/kg or more, with no untoward effect. DDAVP can advantageously replace pitressin tannate in oil as a diagnostic agent.


The removal of the deep lateral wall in orbital decompression: its contribution to exophthalmos reduction and influence on consecutive diplopia.
To evaluate the contribution of maximal removal of the deep lateral wall of the orbit to exophthalmos reduction in Graves' orbitopathy and its influence on the onset of consecutive diplopia. Case-control study. The medical records of two cohorts of patients affected by Graves' orbitopathy with exophthalmos > or = 23 mm, without preoperative diplopia, were retrieved at random from the pool of patients decompressed for rehabilitative reasons at our institution (01/1990 to 12/2003), and retrospectively reviewed. They had been treated with an extended (cases, group 1, n = 15) or conservative (controls, group 2, n = 15) 3-wall orbital decompression performed through a coronal approach. The deep portion of the lateral wall had been removed in the extended decompression group while preserved in the conservative decompression group. Demographics, preoperative characteristics, and surgical outcome were compared. The difference in mean exophthalmos reduction between groups 1 and 2 was considered to be the contribution of the deep lateral wall to reduction of exophthalmos. Groups 1 and 2 were drawn from a pool of 37 and 335 patients, respectively. Demographics and preoperative characteristics of the two groups were not significantly different. The mean contribution of the deep lateral wall to exophthalmos reduction was 2.3 mm. The onset of consecutive diplopia was not significantly different between the two groups (case n = 2/15, controls n = 5/15; P = .203). Diplopia resolved spontaneously in all the patients of group 1, while all the patients of group 2 required surgery. Removal of the deep lateral orbital wall as part of a coronal-approach, 3-wall decompression, enhances the degree of exophthalmos reduction without increasing the risk of consecutive diplopia.


Proton inhibition of chloride exchange: asynchrony of band 3 proton and anion transport sites?
The inhibition of chloride exchange at 0 degrees C by protons at the cytoplasmic and the extracellular surface of the band 3 protein of human erythrocytes was measured between pH 4.6 and 7.6. At constant external pH and chloride concentration, internal protons were a mixed inhibitor of chloride flux, with the apparent pK2 = 6.1 for protonation of the inward-facing empty transporter conformation and the apparent pK3 = 5.7 for protonation of the chloride-transporter complex. The activation of chloride exchange by external chloride was inhibited by internal protons, and internal protonation of the externally facing empty conformation had a pK1 = 6.1. External protons were also a mixed inhibitor of chloride exchange with the apparent pK1 = 5.0 for the empty outward-facing transporter conformation. Because of the pHo dependence of self-inhibition, the value of pK3 on the outside for chloride could not be accurately determined, but the apparent pK3 for protonation of the iodide-transporter complex on the extracellular surface was 4.9. The data support a mechanism with a single proton binding site that can alternatively have access to the cytoplasmic and extracellular solutions. It appears that this proton binding and transport site can be coupled to the single anion transport site for cotransport, but the two sites can be on opposite sides of the membrane at the same time and thus can be asynchronously transported by conformational changes of band 3.


Water dynamics in hectorite clays: influence of temperature studied by coupling neutron spin echo and molecular dynamics.
Within the wider context of water behavior in soils, and with a particular emphasis on clays surrounding underground radioactive waste packages, we present here the translational dynamics of water in clays in low hydrated states as studied by coupling molecular dynamics (MD) simulations and quasielastic neutron scattering experiments by neutron spin echo (NSE). A natural montmorillonite clay of interest is modeled by a synthetic clay which allows us to understand the determining parameters from MD simulations by comparison with the experimental values. We focus on temperatures between 300 and 350 K, i.e., the range relevant to the highlighted application. The activation energy Ea experimentally determined is 6.6 kJ/mol higher than that for bulk water. Simulations are in good agreement with experiments for the relevant set of conditions, and they give more insight into the origin of the observed dynamics.


Control of pepsinogen synthesis and secretion in primary monolayer cultures of canine gastric chief cells.
We have studied pepsinogen synthesis and secretion in primary monolayer cultures of canine gastric chief cells. Monolayers were formed after approximately 48 hr. Pepsinogen synthesis was studied by adding 14C-labeled amino acids to the culture medium. Basal secretion of de novo synthesized pepsinogen after 4 hr was 4 +/- 1.2% of total newly synthesized pepsinogen. Basal secretion of stored pepsinogen after 90 min was 8 +/- 1.4% of total pepsinogen content. Carbachol, dbcAMP, forskolin, VIP, CCK-8, and the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate all stimulated secretion of de novo synthesized pepsinogen and preformed pepsinogen. Only additive interactions were found. dbcAMP caused a peak outburst of stored pepsinogen in the first 10 min. Carbachol stimulation was time dependent. Stimulation of de novo synthesized pepsinogen secretion was time dependent for both carbachol and dbcAMP. dbcAMP caused an immediate 10-fold increase in pepsinogen synthesis, but carbachol did so only after a lag time of 30 min. This was identical to the time necessary for the appearance of labeled pepsinogen in the medium. Addition of atropine after 2 hr resulted in a return to basal synthesis. Stimulated pepsinogen synthesis was always observed concomitant with stimulated pepsinogen secretion. These results show that most external stimuli for pepsinogen synthesis are dependent upon prior depletion of pepsinogen stores, which then triggers synthesis, while stimulation of cAMP production stimulates pepsinogen synthesis more directly.


G-matrix Fourier transform NOESY-based protocol for high-quality protein structure determination.
A protocol for high-quality structure determination based on G-matrix Fourier transform (GFT) NMR is presented. Five through-bond chemical shift correlation experiments providing 4D and 5D spectral information at high digital resolution are performed for resonance assignment. These are combined with a newly implemented (4,3)D GFT NOESY experiment which encodes information of 4D 15N/15N-, 13C(alipahtic)/15N-, and 13C(aliphatic)/13C(aliphatic)-resolved [1H,1H]-NOESY in two subspectra, each containing one component of chemical shift doublets arising from 4D --> 3D projection at omega1:Omega(1H) +/- Omega(X) [X = 15N,13C(aliphatic)]. The peaks located at the centers of the doublets are obtained from simultaneous 3D 15N/13C(aliphatic)/13C(aromatic)-resolved [1H,1H]-NOESY, wherein NOEs detected on aromatic protons are also obtained. The protocol was applied for determining a high-quality structure of the 14 kDa Northeast Structural Genomics consortium target protein, YqfB (PDB ID ). Through-bond correlation and NOESY spectra were acquired, respectively, in 16.9 and 39 h (30 h for shift doublets, 9 h for central peaks) on a 600 MHz spectrometer equipped with a cryogenic probe. The rapidly collected highly resolved 4D NOESY information allows one to assign the majority of NOEs directly from chemical shifts, which yields accurate initial structures "within" approximately 2 angstroms of the final structure. Information theoretical "QUEEN" analysis of initial distance limit constraint networks revealed that, in contrast to structure-based protocols, such NOE assignment is not biased toward identifying additional constraints that tend to be redundant with respect to the available constraint network. The protocol enables rapid NMR data collection for robust high-quality structure determination of proteins up to approximately 20-25 kDa in high-throughput.


Analysis of the Elodea nuttallii transcriptome in response to mercury and cadmium pollution: development of sensitive tools for rapid ecotoxicological testing.
Toxic metals polluting aquatic ecosystems are taken up by inhabitants and accumulate in the food web, affecting species at all trophic levels. It is therefore important to have good tools to assess the level of risk represented by toxic metals in the environment. Macrophytes are potential organisms for the identification of metal-responsive biomarkers but are still underrepresented in ecotoxicology. In the present study, we used next-generation sequencing to investigate the transcriptomic response of Elodea nuttallii exposed to enhanced concentrations of Hg and Cd. We de novo assembled more than 60 000 contigs, of which we found 170 to be regulated dose-dependently by Hg and 212 by Cd. Functional analysis showed that these genes were notably related to energy and metal homeostasis. Expression analysis using nCounter of a subset of genes showed that the gene expression pattern was able to assess toxic metal exposure in complex environmental samples and was more sensitive than other end points (e.g., bioaccumulation, photosynthesis, etc.). In conclusion, we demonstrate the feasibility of using gene expression signatures for the assessment of environmental contamination, using an organism without previous genetic information. This is of interest to ecotoxicology in a wider sense given the possibility to develop specific and sensitive bioassays.


Ultrasound examination before amniocentesis. Its effect on cell culture for cytogenetic studies.
In order to obtain metaphases plates from amniotic fluid cells for chromosome analysis, amniocentesis is performed on patients who are at risk of carrying a fetus with genetic disorders. Ultrasound examination is routinely done before amniocentesis as an aid to the latter procedure and to obtain clinical data concerning the fetus. Speculation that ultrasound examination would reduce the number of blood-stained taps obtained at amniocentesis and maybe also inhibit the growth of amniotic fluid cells in culture is discussed, based on findings in this laboratory over a 3-year period.


Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells.
Brucella species are facultative intracellular pathogens that infect animals as their natural hosts. Transmission to humans is most commonly caused by direct contact with infected animals or by ingestion of contaminated food and can lead to severe chronic infections. Brucella can invade professional and non-professional phagocytic cells and replicates within endoplasmic reticulum (ER)-derived vacuoles. The host factors required for Brucella entry into host cells, avoidance of lysosomal degradation, and replication in the ER-like compartment remain largely unknown. Here we describe two assays to identify host factors involved in Brucella entry and replication in HeLa cells. The protocols describe the use of RNA interference, while alternative screening methods could be applied. The assays are based on the detection of fluorescently labeled bacteria in fluorescently labeled host cells using automated wide-field microscopy. The fluorescent images are analyzed using a standardized image analysis pipeline in CellProfiler which allows single cell-based infection scoring. In the endpoint assay, intracellular replication is measured two days after infection. This allows bacteria to traffic to their replicative niche where proliferation is initiated around 12 hr after bacterial entry. Brucella which have successfully established an intracellular niche will thus have strongly proliferated inside host cells. Since intracellular bacteria will greatly outnumber individual extracellular or intracellular non-replicative bacteria, a strain constitutively expressing GFP can be used. The strong GFP signal is then used to identify infected cells. In contrast, for the entry assay it is essential to differentiate between intracellular and extracellular bacteria. Here, a strain encoding for a tetracycline-inducible GFP is used. Induction of GFP with simultaneous inactivation of extracellular bacteria by gentamicin enables the differentiation between intracellular and extracellular bacteria based on the GFP signal, with only intracellular bacteria being able to express GFP. This allows the robust detection of single intracellular bacteria before intracellular proliferation is initiated.


Amino acids in pediatric and neonatal nutrition.
Modern infant formulas contain sufficient indispensable as well as dispensable amino acids to support normal growth of both term and preterm infants. However, current parenteral and specialized enteral amino acid preparations do not appear to be optimal. Many of the problems with parenteral preparations reflect the fact that ingested amino acids (protein) undergo more extensive enteric and hepatic metabolism than parenterally administered amino acids, including conversion to other amino acids (e.g. arginine) that reach the plasma for support of ongoing protein synthesis. Because this important source of amino acids is bypassed when nutrients are delivered parenterally, parenteral requirements of these amino acids are increased. In addition, while ingested phenylalanine and methionine appear to be converted to tyrosine and cysteine, respectively, it seems that parenterally administered phenylalanine and methionine are not. While glutamine, the branched-chain amino acids and arginine appear to be important in stressed infants and infants with compromised gastrointestinal function, specific roles have not been defined. Finally, because some amino acids are insoluble (e.g. tyrosine) and others are unstable in aqueous solution (e.g. glutamine and cysteine), suitable ways to provide these amino acids are needed. Soluble dipeptides of all types are available and have been shown to be both efficacious and safe in adults. This is also likely to be true for pediatric patients but data concerning their efficacy and safety in this population are lacking. Although recent research has been helpful, further research is necessary to define the optimal amino acid requirements of infants who are dependent on specialized nutrition regimens and to investigate whether these needs may differ depending upon the reasons why parenteral nutrition is required (e.g. growth versus response to stress).


Perforator flaps in late-stage pressure sore treatment: outcome analysis of 11-year-long experience with 143 patients.
In the last decade, perforator flaps have been introduced for the treatment of pressure ulcers as alternative to the more popular myocutaneous local flaps. We reviewed our single-team 11-year experience in order to define whether real advantages could be achieved. We analyzed 143 patients undergoing perforator flap surgery for a single late-stage pressure sore. All patients underwent the same protocol treatment. Data regarding associated pathologies, demographics, complications, healing, and hospitalization times were collected. Ninety-three percent of 143 patients were white Caucasian, and 61% were men, with median age of 51 years. Of 143 stage 4 ulcers, 46.2% were ischial, 42.7% sacral, and 11.2% trochanteric. The most common diagnosis was traumatic paraplegia/tetraplegia (74.9%); no significant difference was found in diagnosis distribution and in ulcer location between recurrent and nonrecurrent patients. We performed 44 S-GAP, 78 I-GAP, 3 PFAP-am, and 18 PFAP-1 flaps. At 2 years' follow-up, the overall recurrence was 22.4% and new ulcer occurrence was 4.2%. Mean hospital stay was 16 days. The overall complication percentage was 22.4%, mostly due to suture-line dehiscence (14%) and distal flap necrosis (6.3%). PFAP flaps had a significant higher risk of developing recurrence than I-GAP flaps. The recurrence risk was significantly higher for subjects suffering from coronary artery disease. Late-stage pressure sore treatment with local perforator flaps can achieve reliable long-term outcomes in terms of recurrences and complications. When compared to previously published data, perforator flaps surgery decreased postoperative hospital stay (by an average of nearly 1 week), reoperations (5.6%), and occurrences.


Nucleotide sequence of the structural gene for tryptophanase of Escherichia coli K-12.
The tryptophanase structural gene, tnaA, of Escherichia coli K-12 was cloned and sequenced. The size, amino acid composition, and sequence of the protein predicted from the nucleotide sequence agree with protein structure data previously acquired by others for the tryptophanase of E. coli B. Physiological data indicated that the region controlling expression of tnaA was present in the cloned segment. Sequence data suggested that a second structural gene of unknown function was located distal to tnaA and may be in the same operon. The pattern of codon usage in tnaA was intermediate between codon usage in four of the ribosomal protein structural genes and the structural genes for three of the tryptophan biosynthetic proteins.


Adaptation of Cost Analysis Studies in Practice Guidelines.
Clinical guidelines play a central role in day-to-day practice. We assessed the degree of incorporation of cost analyses to guidelines and identified modifiable characteristics that could affect the level of incorporation.We selected the 100 most cited guidelines listed on the National Guideline Clearinghouse (http://www.guideline.gov) and determined the number of guidelines that used cost analyses in their reasoning and the overall percentage of incorporation of relevant cost analyses available in PubMed. Differences between medical specialties were also studied. Then, we performed a case-control study using incorporated and not incorporated cost analyses after 1:1 matching by study subject and compared them by the Consolidated Health Economic Evaluation Reporting Standards (CHEERS) statement requirements and other criteria.We found that 57% of guidelines do not use any cost justification. Guidelines incorporate a weighted average of 6.0% (95% confidence interval [CI] 4.3-7.9) among 3396 available cost analyses, with cardiology and infectious diseases guidelines incorporating 10.8% (95% CI 5.3-18.1) and 9.9% (95% CI 3.9- 18.2), respectively, and hematology/oncology and urology guidelines incorporating 4.5% (95% CI 1.6-8.6) and 1.6% (95% CI 0.4-3.5), respectively. Based on the CHEERS requirements, the mean number of items reported by the 148 incorporated cost analyses was 18.6 (SD = 3.7), a small but significant difference over controls (17.8 items; P = 0.02). Included analyses were also more likely to directly relate cost reductions to healthcare outcomes (92.6% vs 81.1%, P = 0.004) and declare the funding source (72.3% vs 53.4%, P < 0.001), while similar number of cases and controls reported a noncommercial funding source (71% vs 72.7%; P = 0.8).Guidelines remain an underused mechanism for the cost-effective allocation of available resources and a minority of practice guidelines incorporates cost analyses utilizing only 6% of the available cost analyses. Fulfilling the CHEERS requirements, directly relating costs with healthcare outcomes and transparently declaring the funding source seem to be valued by guideline-writing committees.


Electron-withdrawing effects on the molecular structure of 2- and 3-nitrobenzonitrile revealed by broadband rotational spectroscopy and their comparison with 4-nitrobenzonitrile.
The rotational spectra of 2- and 3-nitrobenzonitrile were recorded via chirped-pulse Fourier transform microwave spectroscopy in the frequency range of 2-8 GHz. These molecules each display large dipole moments, making them viable candidates for deceleration and trapping experiments with AC-electric fields. For both molecules, the main isotopologues and all isotopologues of the respective 13C-, 15N-, 18O-monosubstituted species in their natural abundance were assigned. These assignments allowed for the structural determination of 2- and 3-nitrobenzonitrile via Kraitchman's equations as well as a mass-dependent least-squares fitting approach. The experimentally determined structural parameters are then compared to those obtained from quantum-chemical calculations for these two molecules and 4-nitrobenzonitrile. Structural changes caused by steric interaction and competition for the electron density of the phenyl ring highlight how these strong electron-withdrawing substituents affect one another according to their respective positions on the phenyl ring.


Laser light visual cueing for freezing of gait in Parkinson disease: A pilot study with male participants.
Freezing of gait (FOG) is a debilitating feature of Parkinson disease (PD). In this pilot study, we sought to assess the efficacy of a rolling walker with a laser beam visual cue to treat FOG in PD patients. We recruited 22 subjects with idiopathic PD who experienced on- and off-medication FOG. Subjects performed three walking tasks both with and without the laser beam while on medications. Outcome measures included time to complete tasks, number of steps, and number of FOG episodes. A crossover design allowed within-group comparisons between the two conditions. No significant differences were observed between the two walking conditions across the three tasks. The laser beam, when applied as a visual cue on a rolling walker, did not diminish FOG in this study.


Lycopene β-cyclase expression influences plant physiology, development, and metabolism in tobacco plants.
Carotenoids are important isoprenoids produced in the plastids of photosynthetic organisms that play key roles in photoprotection and antioxidative processes. β-Carotene is generated from lycopene by lycopene β-cyclase (LCYB). Previously, we demonstrated that the introduction of the Daucus carota (carrot) DcLCYB1 gene into tobacco (cv. Xanthi) resulted in increased levels of abscisic acid (ABA) and especially gibberellins (GAs), resulting in increased plant yield. In order to understand this phenomenon prior to exporting this genetic strategy to crops, we generated tobacco (Nicotiana tabacum cv. Petit Havana) mutants that exhibited a wide range of LCYB expression. Transplastomic plants expressing DcLCYB1 at high levels showed a wild-type-like growth, even though their pigment content was increased and their leaf GA1 content was reduced. RNA interference (RNAi) NtLCYB lines showed different reductions in NtLCYB transcript abundance, correlating with reduced pigment content and plant variegation. Photosynthesis (leaf absorptance, Fv/Fm, and light-saturated capacity of linear electron transport) and plant growth were impaired. Remarkably, drastic changes in phytohormone content also occurred in the RNAi lines. However, external application of phytohormones was not sufficient to rescue these phenotypes, suggesting that altered photosynthetic efficiency might be another important factor explaining their reduced biomass. These results show that LCYB expression influences plant biomass by different mechanisms and suggests thresholds for LCYB expression levels that might be beneficial or detrimental for plant growth.


Metabolic profiling of oxidized lipid-derived volatiles in blood by gas chromatography/mass spectrometry with in-tube extraction.
Once lipids are oxidized, various volatiles are produced by cleavage of the fatty acid side chain. Considering the variety of lipids present in the body, a large number of possible volatiles might originate from oxidized lipids. However, only specific volatiles such as aldehydes are exclusively examined in current studies, and there is no reported method for the exhaustive analysis of all volatiles. We developed a sensitive analytical method for the detection of all possible volatiles for multimarker profiling, applying a new extraction method called in-tube extraction. Oxidized phosphatidyl choline standards were prepared in vitro and analyzed in order to determine the potential variety of volatiles. Over 40 compounds, including alcohols, ketones, and furanones, were identified in addition to the aldehydes reported previously. Based on this result, we applied our analytical method to mouse plasma and identified 12 volatiles, including 1-octen-3-ol, which is correlated to disease states. To determine the volatile profile after oxidation, we oxidized plasma in vitro under various conditions and identified 27 volatiles, including 1-octen-3-ol and benzaldehyde. The generation capacity of each volatile was different. This method allows sensitive and exhaustive analysis of various volatiles in addition to aldehydes.


Novel Multiplex Immunoassays for Quantification of IgG against Group B Streptococcus Capsular Polysaccharides in Human Sera.
Group B Streptococcus (GBS) infections constitute a major cause of invasive disease during the first three months of life and an unmet medical need that could be addressed by maternal vaccination. The GBS capsular polysaccharides (CPSs) have shown promise as vaccine targets in clinical studies. A highly specific serological assay to quantify maternal and neonatal anti-CPS antibody levels will be instrumental for GBS vaccine licensure. Here, we describe the development and comparison of two novel multiplex immunoassays (MIAs) based on the Luminex technology for the quantification of IgG antibodies recognizing the five most frequent GBS capsular variants (Ia, Ib, II, III, and V) out of the ten types identified. The first assay is based on the use of biotinylated CPSs coupled to streptavidin-derivatized magnetic microspheres (Biotin-CPS MIA), while the second is a sandwich assay with plain CPSs coupled to magnetic microspheres coated with polysaccharide-specific mouse monoclonal antibodies (Sandwich MIA). Both assays showed good specificity, linearity, and precision, although the Biotin-CPS MIA presented higher sensitivity and lower complexity than the Sandwich MIA. A panel of human sera representing a wide range of anti-CPS IgG concentrations was tested in parallel by the two assays, which resulted in comparable titers. Our data support the preservation of antigenic epitopes in the biotinylated polysaccharides and the suitability of the Biotin-CPS MIA for the precise determination of GBS anti-CPS IgG concentrations in human sera.IMPORTANCE Group B streptococcal infections can cause death in neonates up to 3 months of age. Intrapartum antibiotic prophylaxis in GBS-colonized mothers has limited early infections but has no impact after the first week of life. The development of a maternal vaccine to address this unmet medical need has been identified as a priority by the World Health Organization, and the GBS CPSs are considered the best antigen targets. However, to date there are no accepted standardized assays to measure immune responses to the investigational vaccines and for establishment of serocorrelates of protection. Here, we describe the performance of two microsphere-based pentaplex immunoassays for the determination of antibodies recognizing the five most frequent GBS serotypes. Our data confirm that an assay based on biotinylated polysaccharides coupled to streptavidin microspheres would be suitable for the intended purpose.