Sensitive detection of BRAF V600E mutation by Amplification Refractory Mutation System (ARMS)-PCR.
BRAF mutations occur in approximately 8% of all human cancers and approach 50% in melanoma and papillary carcinoma of thyroid. These mutations provide potentially valuable diagnostic, prognostic and treatment response prediction markers. A sensitive, specific, low-cost assay to detect these mutations is needed. To detect BRAF V600E mutation in formalin-fixed, paraffin-embedded (FFPE) tissue, we developed a method using Amplification Refractory Mutation System (ARMS)-PCR. This method was designed to amplify three products in a single reaction tube: a 200 bp common product serving as an amplification control, a 144 bp BRAF V600E specific product, and a 97 bp wild-type (wt) specific product. The sensitivity of this method was determined to be as low as 0.5% for the BRAF V600E allele in a wild-type background. This method was successfully validated in 72 thyroid tumors. It detected V600E mutation in 22 out of 33 (67%) of the conventional papillary thyroid carcinoma (PTC), 8 out of 12 (75%) of the tall-cell variant of PTC, whereas none of the 10 follicular variant of PTC showed BRAF V600E mutation. In addition, none of the 14 follicular adenomas and 3 follicular carcinomas had BRAF V600E mutation. As a comparison method, direct dideoxy sequencing found only 27 out of 30 (90%) mutations detected by ARMS-PCR method, suggesting that this ARMS-PCR method has higher sensitivity. Our ARMS-PCR method provides a new tool for rapid detection of BRAF V600E mutation. Our results indicate that ARMS-PCR is more sensitive than automated dideoxy sequencing in detecting low BRAF V600E allele burdens in FFPE tumor specimen. The strategy of this ARMS-PCR design may be adapted for early detection of point mutations of a variety of biomarker genes.


Pushing functional MRI spatial and temporal resolution further: High-density receive arrays combined with shot-selective 2D CAIPIRINHA for 3D echo-planar imaging at 7 T.
To be able to examine dynamic and detailed brain functions, the spatial and temporal resolution of 7 T MRI needs to improve. In this study, it was investigated whether submillimeter multishot 3D EPI fMRI scans, acquired with high-density receive arrays, can benefit from a 2D CAIPIRINHA sampling pattern, in terms of noise amplification (g-factor), temporal SNR and fMRI sensitivity. High-density receive arrays were combined with a shot-selective 2D CAIPIRINHA implementation for multishot 3D EPI sequences at 7 T. In this implementation, in contrast to conventional inclusion of extra kz gradient blips, specific EPI shots are left out to create a CAIPIRINHA shift and reduction of scan time. First, the implementation of the CAIPIRINHA sequence was evaluated with a standard receive setup by acquiring submillimeter whole brain T2 *-weighted anatomy images. Second, the CAIPIRINHA sequence was combined with high-density receive arrays to push the temporal resolution of submillimeter 3D EPI fMRI scans of the visual cortex. Results show that the shot-selective 2D CAIPIRINHA sequence enables a reduction in scan time for 0.5 mm isotropic 3D EPI T2 *-weighted anatomy scans by a factor of 4 compared with earlier reports. The use of the 2D CAIPIRINHA implementation in combination with high-density receive arrays, enhances the image quality of submillimeter 3D EPI scans of the visual cortex at high acceleration as compared to conventional SENSE. Both the g-factor and temporal SNR improved, resulting in a method that is more sensitive to the fMRI signal. Using this method, it is possible to acquire submillimeter single volume 3D EPI scans of the visual cortex in a subsecond timeframe. Overall, high-density receive arrays in combination with shot-selective 2D CAIPIRINHA for 3D EPI scans prove to be valuable for reducing the scan time of submillimeter MRI acquisitions.


"Ion-Flux": The natural course of ECG changes in severe hypokalemia.
Whereas the electrocardiogram (ECG) changes in hypokalemia are well known, they often receive less attention than the more striking features of hyperkalemia. Furthermore, there is a need for further discussion as to the subtleties of ECG changes that can aid in the differential diagnoses. This case study presents the ECG changes of a patient with severe hypokalemia due to diarrhea. It highlights how bifid T-waves in hypokalemia can be distinguished from other conditions such as coronary artery disease or pericarditis. Furthermore, it also shows the gradual reversal of ECG changes in the same patient when potassium is normalized.


Nonaggregating insulin solutions for long-term glucose control in experimental and human diabetes.
A physiologic additive for dissolving insulin crystals for parenteral application has been found. Insulin crystals are relatively insoluble in simple aqueous solutions. They will dissolve, however, in highly acidic solutions, but these are not suitable for parenteral use. Both neutral and acid pH insulin solutions have a tendency for the dissolved hormone to reaggregate. Notwithstanding possible changes in biologic activity, such formed aggregates must be prevented because they interfere with the flow in portable insulin delivery devices and result in the loss of glycemic control. The addition of 1.5% autologous serum to the aqueous diluent for insulin has eliminated these difficulties and increased by 37% the apparent biologic activity of insulin solutions prepared in this way. With this additive, continuous uninterrupted intravenous insulin infusion has provided near ideal blood glucose control in four pancreatectomized dogs for 5 mo and four patients with juvenile-onset diabetes for 18--23 days. Serum apparently contains factor(s) that promote the dissolution of insulin and prevent the formation of peptide aggregates in dilute solutions.


Telemedicine for gastrointestinal endoscopy: The Endoscopic Club E-conference in the Asia Pacific Region.
Background and study aims An Endoscopic Club E-conference (ECE) was set up in May 2014 to cater to increased demand for gastrointestinal endoscopy-related teleconferences in the Asia-Pacific region which were traditionally organized by the medical working group (MWG) of Asia-Pacific Advanced Network. This study describes how the ECE meeting was run, examines the group dynamics, outlines feedback and analyzes factors affecting the enthusiasm of participants. It is hoped that the findings here can serve as guidance for future development of other teleconference groups. Methods The preparation, running of and feedback on the ECE teleconference were evaluated and described. The country's economic situation, time zone differences, connectivity with a research and education network (REN) and engineering cooperation of each member were recorded and analyzed with regard to their association with participant enthusiasm, which was taken as participation in at least 50 % of the meetings since joining. Associations were calculated using 2-way table with chi-square test to generate odds ratio and P value. Results To date, ECE members have increased from 7 to 29 (increment of 314 %). Feedback received indicated a high level of satisfaction with program content, audiovisual transmission and ease of technical preparation. Upper gastrointestinal luminal endoscopy-related topics were the most favored program content. Those topics were presented mainly via case studies with a focus on management challenges. Time zone differences of more than 6 hours and poor engineering cooperation were independently associated with inactive participation (P values of 0.04 and 0.001 respectively). Conclusions Good program content and high-quality audiovisual transmission are keys to the success of an endoscopic medical teleconference. In our analysis, poor engineering cooperation and discordant time zones contributed to inactive participation while connectivity with REN and a country's economic situation were not significantly associated with participant enthusiasm.


Performance tests in people with chronic low back pain: responsiveness and minimal clinically important change.
Cohort study. To assess the responsiveness and minimal clinically important change (MCIC) of 6 commonly-used performance tests (5-minute walking, 50-ft walking, sit-to-stand, 1 minute stair climbing, loaded forward reach, Progressive Isoinertial Lifting Evaluation). Performance tests are used to evaluate physical function in people with low back pain. Little is known about their clinimetric properties. Performance tests were administered in people with chronic nonspecific low back pain (n = 198) before and after 10 weeks of treatment. At 10 weeks, the global perceived effect scale was used to determine if participants judged themselves as worsened, unchanged, or improved. The mean change scores for each performance test were calculated. A performance test was considered responsive if the area under the receiver operating characteristic curve (AUC) was equal to or greater than 0.70. We used 2 methods to evaluate MCIC: the optimal cut-off point based on the receiver operating characteristic curve, which takes into account both sensitivity and specificity, and the minimal detectable change for improvement, which considers test specificity only. In general, the mean change scores were the smallest in participants who judged themselves worsened and largest in those reporting to be improved. Sit-to-stand (AUC = 0.75) and stair climbing (AUC = 0.72) were the only performance tests that showed adequate responsiveness. For sit-to-stand, the MCIC ranged from 4.1 to 9.8 seconds (19%-45% of the mean baseline score). For stair climbing, the MCIC ranged from 14.5 to 23.9 steps (19%-31% of the mean baseline score). Only 2 of the 6 performance tests were responsive. Both had acceptable MCIC values. Developing individualized performance tests might partly overcome the general lack of responsiveness of performance tests. Future research should focus on the clinimetric testing of performance tests in subgroups.


Mechanisms behind the failure of residents' longitudinal primary care to promote disclosure and discussion of psychosocial issues.
Longitudinality (care by a single physician over time) and continuity (receipt of most care from a single physician) are believed to enhance patient-physician relationships and facilitate disclosure of emotional distress, but some studies suggest this potential goes unrealized. To determine whether care in a pediatric residents' continuity clinic promotes, over time, increased discussion, disclosure, and detection of parents' social and emotional distress and to understand physicians' communication behaviors underlying changes with time. Longitudinal, observational study of parent-physician interaction over the course of 1 year. One hundred ninety parents (90% African American) and their infants' primary care physicians (31 [4 Asians and 27 whites] first- and second-year pediatric residents). Frequency with which parents and physicians raised topics related to parental mood and family or social functioning; proportion of distressed parents discussing mood or functioning; and physicians' detection of parent distress. Physician initiation of psychosocial topics fell in the course of longitudinal relationships (odds of initiation in visits > or =6 vs odds of initiation in visits 1-5 = 0.46 [95% confidence limits, 0.31%, 0.67%]); parent initiation did not change over time nor was it increased by greater levels of continuity. Length of relationship was not associated with increased physician detection of parental distress or with increased rates of disclosure by distressed parents. Physicians' positively framed leading questions, and their avoidant responses to prior parental disclosures were significantly associated with decreased odds of problem disclosure. In contrast, visits in which parents or physicians raised psychosocial topics were characterized, on average, by 40% higher levels of physicians' "patient-centeredness" (increases of about 100 utterances per visit [95% confidence limits, 65.7%, 133.9%]). Longitudinal relationships between residents and patients may not be sufficient to promote the discussion, disclosure, and detection of psychosocial issues. Training in communication skills may help residents achieve the potential and goals of longitudinal care.


Serum carotenoid interactions in premenopausal women reveal α-carotene is negatively impacted by body fat.
Increasing body mass indices (BMIs) across the globe reflect pandemic shifts towards habitual positive energy imbalances. Excess body fat in individuals is often associated with high-energy and high-fat diets scanty in fresh produce. Carotenoids are fat-soluble pigments plentiful in many fruits and vegetables. They are well-known for provitamin A and antioxidant functions, but little research has been done related to carotenoid-body mass interactions. Serum carotenoids were analyzed relative to body fat to determine correlations between major serum carotenoids, retinol, BMI, fat mass, and lean mass. Healthy women ( n = 76), 19-50 years old, were categorized into two comparison groups determined by percent body fat measured by air displacement plethysomography (BOD POD®), i.e. <31% and ≥31% fat mass. Anthropometric and three-day diet records were completed for BMI and nutrient intake calculations, respectively. Serum α-carotene concentrations were strongly inversely associated with all measures of body composition ( P < 0.001 α-carotene) controlling for dietary intake and age, while β-carotene, lutein, and lycopene were not ( P > 0.05). Dietary intake between groups did not differ, including carrot consumption (a high dietary source of α-carotene). These results confirm previous carotenoid-health research and propose the need for further investigation of potential protective roles that α-carotene may perform for optimal health. Serum α-carotene may provide a deeper and clinically relevant purpose, beyond previous suggestions for its use as a biomarker for fruit and vegetable consumption, in that α-carotene may be a biomarker for chronic disease risk frequently linked with obesity. Impact statement Carotenoids are important pigments in fruit and vegetables and found in human serum. This study isolated a negative relationship between serum α-carotene and body fatness. As humans begin to live over a century, determining biomarkers of ultimate health is important. α-Carotene does not have the same distribution in the food supply as β-carotene and therefore is often overlooked in surveys. In part, this is due to the fact that β-carotene provides two molecules of vitamin A, while α-carotene provides one upon central cleavage. This study shows a very clear association between α-carotene and body fatness, which appears to go beyond its fat-soluble nature. Dietary intake data were not able to explain the association. Further work is needed to determine what dietary components infer health benefits.


Pharmacophore based virtual screening, molecular docking, molecular dynamics and MM-GBSA approach for identification of prospective SARS-CoV-2 inhibitor from natural product databases.
COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) primarily appeared in Wuhan, China, in December 2019. At present, no proper therapy and vaccinations are available for the disease, and it is increasing day by day with a high mortality rate. Pharmacophore based virtual screening of the selected natural product databases followed by Glide molecular docking and dynamics studies against SARS-CoV-2 main protease was investigated to identify potential ligands that may act as inhibitors. The molecules SN00293542 and SN00382835 revealed the highest docking score of -14.57 and -12.42 kcal/mol, respectively, when compared with the co-crystal ligands of PDB-6Y2F (O6K) and 6W63 (X77) of the SARS-CoV-2 Mpro. To further validate the interactions of top scored molecules SN00293542 and SN00382835, molecular dynamics study of 100 ns was carried out. This indicated that the protein-ligand complex was stable throughout the simulation period, and minimal backbone fluctuations have ensued in the system. Post-MM-GBSA analysis of molecular dynamics data showed free binding energy-71.7004 +/- 7.98, -56.81+/- 7.54 kcal/mol, respectively. The computational study identified several ligands that may act as potential inhibitors of SARS-CoV-2 Mpro. The top-ranked molecules SN00293542, and SN00382835 occupied the active site of the target, the main protease like that of the co-crystal ligand. These molecules may emerge as a promising ligands against SARS-CoV-2 and thus needs further detailed investigations. Communicated by Ramaswamy H. Sarma.


Respiratory function in patients post-infection by COVID-19: a systematic review and meta-analysis.
Evidence suggests lungs as the organ most affected by coronavirus disease 2019 (COVID-19). The literature on previous coronavirus infections reports that patients may experience persistent impairment in respiratory function after being discharged. Our objective was to determine the prevalence of restrictive pattern, obstructive pattern and altered diffusion in patients post-COVID-19 infection and to describe the different evaluations of respiratory function used with these patients. A systematic review was conducted in five databases. Studies that used lung function testing to assess post-infection COVID-19 patients were included for review. Two independent reviewers analysed the studies, extracted the data and assessed the quality of evidence. Of the 1973 reports returned by the initial search, seven articles reporting on 380 patients were included in the data synthesis. In the sensitivity analysis, we found a prevalence of 0.39 (CI 0.24-0.56, p < 0.01, I2 = 86%), 0.15 (CI 0.09-0.22, p = 0.03, I2 = 59%), and 0.07 (CI 0.04-0.11, p = 0.31, I2 = 16%) for altered diffusion capacity of the lungs for carbon monoxide (DLCO), restrictive pattern and obstructive pattern, respectively. Post-infection COVID-19 patients showed impaired lung function; the most important of the pulmonary function tests affected was the diffusion capacity.


Anaesthesia, not number of sessions, influences the magnitude and duration of an aHF-rTMS in dogs.
Currently, the rat has been a useful animal model in brain stimulation research. Nevertheless, extrapolating results from rodent repetitive Transcranial Magnetic Stimulation (rTMS) research to humans contains several hurdles. This suggests the desperate need for a large animal model in translational rTMS research. The dog would be a valid choice, not only due to the fact that humans and dogs share a neurophysiological background, but a similar neuropathological background as well. In order to evaluate the feasibility of the canine rTMS animal model, this study aimed to evaluate the neurophysiological response in dogs on a, clinically used, accelerated high frequency (aHF) rTMS protocol. This aHF-rTMS (20 Hz) protocol was performed under anaesthesia or sedation and either 20 sessions or 5 sessions were given to each dog. 21 healthy dogs were randomly subjected to one of the four aHF-rTMS protocols (1 sham and 3 active protocols). For each dog, the perfusion indices (PI), of a [99mTc]HMPAO scan at 4 time points, for the left frontal cortex (stimulation target) were calculated for each protocol. Concerning sham stimulation, the average PI remained at the baseline level. The main result was the presence of a direct transitory increase in rCBF at the stimulation site, both under anaesthesia and sedation. Nevertheless the measured increase in rCBF was higher but shorter duration under sedation. The magnitude of this increase was not influenced by number of sessions. No changes in rCBF were found in remote brain regions. This study shows that, despite the influence of anaesthesia and sedation, comparable and clinically relevant effects on the rCBF can be obtained in dogs. Since less methodological hurdles have to be overcome and comparable results can be obtained, it would be acceptable to put the dog forward as an alternative translational rTMS animal model.


Multifunctional properties of phosphate-solubilizing microorganisms grown on agro-industrial wastes in fermentation and soil conditions.
One of the most studied approaches in solubilization of insoluble phosphates is the biological treatment of rock phosphates. In recent years, various techniques for rock phosphate solubilization have been proposed, with increasing emphasis on application of P-solubilizing microorganisms. The P-solubilizing activity is determined by the microbial biochemical ability to produce and release metabolites with metal-chelating functions. In a number of studies, we have shown that agro-industrial wastes can be efficiently used as substrates in solubilization of phosphate rocks. These processes were carried out employing various technologies including solid-state and submerged fermentations including immobilized cells. The review paper deals critically with several novel trends in exploring various properties of the above microbial/agro-wastes/rock phosphate systems. The major idea is to describe how a single P-solubilizing microorganism manifests wide range of metabolic abilities in different environments. In fermentation conditions, P-solubilizing microorganisms were found to produce various enzymes, siderophores, and plant hormones. Further introduction of the resulting biotechnological products into soil-plant systems resulted in significantly higher plant growth, enhanced soil properties, and biological (including biocontrol) activity. Application of these bio-products in bioremediation of disturbed (heavy metal contaminated and desertified) soils is based on another important part of their multifunctional properties.


Cardiac cine imaging at 3 Tesla: initial experience with a 32-element body-array coil.
We sought to assess the feasibility of cardiac cine imaging and evaluate image quality at 3 T using a body-array coil with 32 coil elements. Eight healthy volunteers (3 men; median age 29 years) were examined on a 3-T magnetic resonance scanner (Magnetom Trio, Siemens Medical Solutions) using a 32-element phased-array coil (prototype from In vivo Corp.). Gradient-recalled-echo (GRE) cine (GRAPPAx3), GRE cine with tagging lines, steady-state-free-precession (SSFP) cine (GRAPPAx3 and x4), and SSFP cine(TSENSEx4 andx6) images were acquired in short-axis and 4-chamber view. Reference images with identical scan parameters were acquired using the total-imaging-matrix (Tim) coil system with a total of 12 coil elements. Images were assessed by 2 observers in a consensus reading with regard to image quality, noise and presence of artifacts. Furthermore, signal-to-noise values were determined in phantom measurements. In phantom measurements signal-to-noise values were increased by 115-155% for the various cine sequences using the 32-element coil. Scoring of image quality yielded statistically significant increased image quality with the SSFP-GRAPPAx4, SSFP-TSENSEx4, and SSFP-TSENSEx6 sequence using the 32-element coil (P < 0.05). Similarly, scoring of image noise yielded a statistically significant lower noise rating with the SSFP-GRAPPAx4, GRE-GRAPPAx3, SSFP-TSENSEx4, and SSFP-TSENSEx6 sequence using the 32-element coil (P < 0.05). This study shows that cardiac cine imaging at 3 T using a 32-element body-array coil is feasible in healthy volunteers. Using a large number of coil elements with a favorable sensitivity profile supports faster image acquisition, with high diagnostic image quality even for high parallel imaging factors.


Images, words, and questions: variables that influence beliefs about vision in children and adults.
In three studies we used animated computer graphics to examine beliefs among children and adults that vision involved input to the eyes (the intromission theory) or emissions from the eye (the extramission theory). Results supported previous findings which showed a decrease in extramission and an increase in intromission responses across age. The findings also indicated that there were more extramission interpretations when subjects were tested with graphic images, and more intromission interpretations when the questioning was purely verbal. However, the magnitude of the effect was highly dependent upon question format. The differences between graphic and verbal question presentations (A) are consistent with our theory on the origins of extramission beliefs, (B) suggest that beliefs can vary as a function of form of symbolization, and (C) are contrary to long-standing beliefs of educators and psychologists that emphasize the importance of concrete, pictorial representation.


Quantification of specific T and B cells immunological markers in children with chronic and transient ITP.
Immune thrombocytopenic purpura (ITP) is characterized by a transient (nonchronic) or permanent (chronic) decline in the number of platelets. Predicting the course of ITP, at the time of diagnosis, is of importance. Here we studied at diagnosis, clinical and immunological parameters in order to distinguish between different courses. The latter included the measure of new B and T cells using quantification of kappa-deleting recombination excision circles (KRECs) and T-cell receptor excision circles (TRECs), respectively. Blood samples were collected from 44 children with a clinical diagnosis of ITP. Real-time PCR was performed in order to quantify the number of copies of TREC and KREC followed by collection of clinical data from medical files. The children were retrospectively divided into two groups: chronic and nonchronic. Twenty-four patients (54%) were classified as nonchronic ITP and 20 patients (46%) were classified as chronic ITP. We confirmed some clinical parameters (e.g., gender, age) but not others (e.g., preceding infection, level of thrombocytopenia) that distinguish patients with chronic and nonchronic course. While KREC quantification was similar in patients regardless the outcome of their disease, it was significantly higher than the level of controls (P < 0.05). TREC quantification was not different between patients and controls. KREC but not TREC levels are different in patients comparing to controls, pointing to an overreaction of B-cell development as a role in the pathogenesis of ITP. These results may shed more lights on the immune mechanism of ITP.


Export of a heterologous cytochrome P450 (CYP105D1) in Escherichia coli is associated with periplasmic accumulation of uroporphyrin.
This report suggests an important physiological role of a CYP in the accumulation of uroporphyrin I arising from catalytic oxidative conversion of uroporphyrinogen I to uroporphyrin I in the periplasm of Escherichia coli cultured in the presence of 5-aminolevulinic acid. A structurally competent Streptomyces griseus CYP105D1 was expressed as an engineered, exportable form in aerobically grown E. coli. Its progressive induction in the presence of 5-aminolevulinic acid-supplemented medium was accompanied by an accumulation of a greater than 100-fold higher amount of uroporphyrin I in the periplasm relative to cells lacking CYP105D1. Expression of a cytoplasm-resident engineered CYP105D1 at a comparative level to the secreted form was far less effective in promoting porphyrin accumulation in the periplasm. Expression at a 10-fold molar excess over the exported CYP105D1 of another periplasmically exported hemoprotein, the globular core of cytochrome b5, did not substitute the role of the periplasmically localized CYP105D1 in promoting porphyrin production. This, therefore, eliminated the possibility that uroporphyrin accumulation is merely a result of increased hemoprotein synthesis. Moreover, in the strain that secreted CYP105D1, uroporphyrin production was considerably reduced by azole-based P450 inhibitors. Production of both holo-CYP105D1 and uroporphyrin was dependent upon 5-aminolevulinic acid, except that at higher concentrations this resulted in a decrease in uroporphyrin. This study suggests that the exported CYP105D1 oxidatively catalyzes periplasmic conversion of uroporphyrinogen I to uroporphyrin I in E. coli. The findings have significant implications in the ontogenesis of human uroporphyria-related diseases.


Bacterial disease and antimicrobial susceptibility patterns in HIV-infected, hospitalized children: a retrospective cohort study.
Serious bacterial infections are a major source of morbidity and mortality in HIV-infected children. The spectrum of disease is wide, and responsible organisms vary according to setting. The use of antibiotic prophylaxis and the emergence of multi-drug resistant bacteria necessitate examination of responsible organisms and their antibiotic susceptibility. A retrospective cohort study of all HIV-positive pediatric admissions at an urban public sector hospital in Cape Town between January 2002 and June 2006 was conducted. Children between the ages of one month and nine years with laboratory confirmed HIV status, serious bacterial infection, and a hospital length of stay of 5 days or more, were eligible for inclusion. Organisms isolated from blood, urine, and cerebral spinal fluid cultures and their antimicrobial susceptibility were examined, and compared according to timing of isolation to distinguish nosocomial versus community-acquired. One hundred and forty-one children were identified (median age 1.2 years), 39% of whom were on antiretrovirals started before or during this hospitalization. Bacterial infections involved all organ systems, however pneumonia was most common (67%). S. pneumoniae and S. aureus were the most common gram positive and K. pneumoniae was the most common gram negative organism. K pneumoniae isolates were resistant to many first and second line antibiotics, and were all considered nosocomial. All S. aureus isolates were methicillin resistant, some of which were community-acquired. Bacterial infections are an important source of co-morbidity in HIV-infected children in resource-limited settings. Clinicians should have a low threshold to initiate antibiotics in children requiring hospitalization. Broad-spectrum antibiotics should be used judiciously. Clinicians caring for HIV-infected children should be cognizant of the most common organisms affecting such children, and of their local antimicrobial susceptibilities, when treating empirically for serious bacterial infections.


Recombinant human erythropoietin promotes the acquisition of a malignant phenotype in head and neck squamous cell carcinoma cell lines in vitro.
Recent studies indicate an increase in tumor progression and recurrence in head and neck squamous cell carcinomas (HNSCC) of cancer patients taking recombinant human erythropoietin (rhEpo) for anemia. This study was undertaken to investigate the potential role of rhEpo in invasion, proliferation, and cisplatin-induced cell death in HNSCC cell lines. The following experiments were performed with two HNSCC cell lines, UMSCC-10B and UMSCC-22B. Presence of EpoR in both cell lines was determined by western blot and quantitative PCR. Colorimetric MTS assays and clonogenic assays were used to study the effect of rhEpo at pharmacologically relevant doses on cell proliferation. Matrigel invasion assays were performed in order to determine effects of exogenous rhEpo on invasive abilities. Clonogenic assays were also used to study potential cytoprotective effects of rhEpo against cisplatin. Immunoblotting was done to analyze the effect of rhEpo on Akt phosphorylation. Finally, MTS and TUNEL assays were performed to test our hypothesis that Akt activation by PI3K was involved in rhEpo-mediated cisplatin resistance. HNSCC cell lines were shown to express Epo receptor (EpoR). RhEpo increased invasion 1.8-fold in UMSCC-10B and 2.6-fold in UMSCC-22B compared to control. RhEpo at 10 U/ml increased cell proliferation by 41% and 53% in UMSCC-10B and UMSCC-22B, respectively, and colony formation by 1.5-fold and 1.8-fold. UMSCC-10B treated with cisplatin and exposed to rhEpo at 1 and 10 U/ml resulted in a 1.7-fold and 3.0-fold increase in colony number compared to control, respectively. UMSCC-22B treated with cisplatin and rhEpo at 1 or 10 U/ml resulted in ~2.5-fold increase in colony number. A TUNEL assay demonstrated a 30.5% and 76.5% increase in survival in UMSCC-10B and UMSCC-22B cells, respectively, in cisplatin and rhEpo-treated cells compared to cisplatin alone. MTS assay showed similar cytoprotective effects. Western blot revealed increased phosphorylation of Akt upon exposure of HNSCC cell lines to rhEpo. MTS assay and TUNEL analyses implicate Akt as a likely contributor to regulation of rhEpo-mediated cytoprotection. The results demonstrate that, in HNSCC cells expressing functional EpoR, rhEpo promotes invasion, cell proliferation, and induces resistance to cisplatin, which may contribute to tumor progression.


Increasing the distance between the snRNA promoter and the 3' box decreases the efficiency of snRNA 3'-end formation.
Chimeric genes which contained the mouse U1b snRNA promoter, portions of the histone H2a or globin coding regions and the U1b 3'-end followed by a histone 3'-end were constructed. The distance between the U1 promoter and the U1 3' box was varied between 146 and 670 nt. The chimeric genes were introduced into CHO cells by stable transfection or into Xenopus oocytes by microinjection. The efficiency of utilization of the U1 3' box, as measured by the relative amounts of transcripts that ended at the U1 3' box and the histone 3'-end, was dependent on the distance between the promoter and 3'-end box. U1 3'-ends were formed with >90% efficiency on transcripts shorter than 200 nt, with 50-70% efficiency on transcripts of 280-400 nt and with only 10-20% efficiency on transcripts >500 nt. Essentially identical results were obtained after stable transfection of CHO cells or after injecting the genes into Xenopus oocytes. The distance between the U1 promoter and the U1 3' box must be <280 nt for efficient transcription termination at the U1 3' box, regardless of the sequence transcribed.


Myosin subunits and contractile properties of single fibers from hypokinetic rat muscles.
The effects of prolonged hypokinesia on the contractile properties and myosin isozymes of single fibers from the synergistic fast-twitch plantaris (PL) and slow-twitch soleus (SOL) skeletal muscles of adult rats were studied after 28 days of hindlimb suspension. There was a 31% increase in the mean maximal velocity of unloaded shortening (Vmax) among fibers from SOL with no change in the mean Vmax of fibers from PL after suspension. The myosin heavy and light chain (MHC and MLC) composition of bundles and the MHC composition of single fibers from control and suspended muscles were examined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. There was a marked increase in the relative amount of fast-type MHC's in hypokinetic SOL and a smaller increase in the amount of fast-type MHC's in the PL. Relatively minor changes occurred in the MLC's during hypokinesia. As Vmax increased among individual fibers from control and suspended muscles, the relative amount of fast-type MHC's increased. The results demonstrate that the myosin isozyme composition of skeletal muscle, especially the heavy chains, is altered during hypokinesia, and this finding provides an explanation for changes in Vmax of rat single muscle fibers under the same conditions.


Radical formation in lithium formate EPR dosimeters after irradiation with protons and nitrogen ions.
Radical formation in polycrystalline lithium formate monohydrate after irradiation with gamma rays, protons and nitrogen ions at room temperature was studied by continuous-wave electron paramagnetic resonance (EPR) spectroscopy. The linear energy transfer (LET) of the various radiation beams was 0.2, 0.7-3.9 and 110-164 keV/microm for gamma rays, protons and nitrogen ions, respectively. Doses between 5 and 20 Gy were given. The EPR reading (the area under the EPR absorption resonance) increased linearly with dose for all types of radiation. As the LET increased, the relative effectiveness (the EPR reading per dose relative to that for gamma rays) decreased, while the EPR line width increased. Track structure theory and modeling of detector effectiveness predicted the dosimeter response observed after proton and nitrogen-ion irradiation. A semi-empirical line broadening model including dipolar spin-spin interactions was developed that explained the dependence of the line width on LET. The findings indicate that the local radical density in lithium formate is increased after high-LET irradiation.


Pulmonary langerhans cell histiocytosis.
Pulmonary Langerhans Cell Histiocytosis (PLCH) is a relatively uncommon lung disease that generally, but not invariably, occurs in cigarette smokers. The pathologic hallmark of PLCH is the accumulation of Langerhans and other inflammatory cells in small airways, resulting in the formation of nodular inflammatory lesions. While the overwhelming majority of patients are smokers, mechanisms by which smoking induces this disease are not known, but likely involve a combination of events resulting in enhanced recruitment and activation of Langerhans cells in small airways. Bronchiolar inflammation may be accompanied by variable lung interstitial and vascular involvement. While cellular inflammation is prominent in early disease, more advanced stages are characterized by cystic lung destruction, cicatricial scarring of airways, and pulmonary vascular remodeling. Pulmonary function is frequently abnormal at presentation. Imaging of the chest with high resolution chest CT scanning may show characteristic nodular and cystic abnormalities. Lung biopsy is necessary for a definitive diagnosis, although may not be required in instances were imaging findings are highly characteristic. There is no general consensus regarding the role of immunosuppressive therapy in smokers with PLCH. All smokers must be counseled on the importance of smoking cessation, which may result in regression of disease and obviate the need for systemic immunosuppressive therapy. The prognosis for most patients is relatively good, particularly if longitudinal lung function testing shows stability. Complications like pneumothoraces and secondary pulmonary hypertension may shorten life expectancy. Patients with progressive disease may require lung transplantation.


Acute Kynurenine Challenge Disrupts Sleep-Wake Architecture and Impairs Contextual Memory in Adult Rats.
Tryptophan metabolism via the kynurenine pathway may represent a key molecular link between sleep loss and cognitive dysfunction. Modest increases in the kynurenine pathway metabolite kynurenic acid (KYNA), which acts as an antagonist at N-methyl-d-aspartate and α7 nicotinic acetylcholine receptors in the brain, result in cognitive impairments. As glutamatergic and cholinergic neurotransmissions are critically involved in modulation of sleep, our current experiments tested the hypothesis that elevated KYNA adversely impacts sleep quality. Adult male Wistar rats were treated with vehicle (saline) and kynurenine (25, 50, 100, and 250 mg/kg), the direct bioprecursor of KYNA, intraperitoneally at zeitgeber time (ZT) 0 to rapidly increase brain KYNA. Levels of KYNA in the brainstem, cortex, and hippocampus were determined at ZT 0, ZT 2, and ZT 4, respectively. Analyses of vigilance state-related parameters categorized as wake, rapid eye movement (REM), and non-REM (NREM) as well as spectra power analysis during NREM and REM were assessed during the light phase. Separate animals were tested in the passive avoidance paradigm, testing contextual memory. When KYNA levels were elevated in the brain, total REM duration was reduced and total wake duration was increased. REM and wake architecture, assessed as number of vigilance state bouts and average duration of each bout, and theta power during REM were significantly impacted. Kynurenine challenge impaired performance in the hippocampal-dependent contextual memory task. Our results introduce kynurenine pathway metabolism and formation of KYNA as a novel molecular target contributing to sleep disruptions and cognitive impairments.


Elastic intramedullary nailing of midclavicular fractures.
The majority of midclavicular fractures are treated conservatively. Fractures that require internal fixation are treated by plate osteosynthesis as the standard procedure. Elastic stable intramedullary nailing is an increasingly popular alternative for the internal fixation of displaced midclavicular fractures. In a three-year period, 15 fractures of the clavicle were treated by elastic intramedullary nailing. Fracture healing, clinical outcome and complications were assessed. Mean follow-up time was one year All fractures healed clinically and radiologically. Non-union or infections were not observed. Functional results according to the Constant score were excellent. Complications consisted of skin irritation in four cases. One acromioclavicular joint separation was observed postoperatively. Elastic stable intramedullary nailing of displaced midclavicular fractures is a minimally invasive technique with excellent functional results and early pain relief.


[Vitrectomy without scleral buckle versus ab-externo approach for pseudophakic retinal detachment: comparative retrospective study].
Retinal detachment (RD) is a major complication of cataract surgery, which can be treated by either primary vitrectomy without indentation or the scleral buckling procedure. The aim of this study is to compare the results of these two techniques for the treatment of pseudophakic RD. The charts of 40 patients (40 eyes) treated with scleral buckling for a primary pseudophakic RD were retrospectively studied and compared to the charts of 32 patients (32 eyes) treated with primary vitrectomy without scleral buckle during the same period by the same surgeons. To obtain comparable samples, patients with giant retinal tears, vitreous hemorrhage, and severe preoperative proliferative vitreoretinopathy (PVR) were not included. Minimal follow-up was 6 months. The primary success rate was 84% in the vitrectomy group and 82.5% in the ab-externo group. Final anatomical success was observed in 100% of cases in the vitrectomy group and in 95% of cases in the ab-externo group. Final visual acuity was 0.5 or better in 44% of cases in the vitrectomy group and 37.5% in the ab-externo group. The duration of the surgery was significantly lower in the ab-externo group, whereas the hospital stay tended to be lower in the vitrectomy group. In the vitrectomy group, postoperative PVR developed in 3 eyes and new or undetected breaks were responsible for failure of the initial procedure in 2 eyes. Primary vitrectomy appears to be as effective as scleral buckling procedures for the treatment of pseudophakic RD.


Lyme disease: a selective medium for isolation of the suspected etiological agent, a spirochete.
A simple procedure with a new selective culture medium for the isolation of the suspected etiological agent of Lyme disease from ticks is described. Live ticks (Ixodes dammini) were ground with a mortar and pestle, and the suspensions were inoculated into a selective and nonselective medium. The selective medium, which contained kanamycin and 5-fluorouracil, yielded positive spirochete cultures from 100% of the pooled ticks and from 79% of the single tick specimens. The isolation rate for the nonselective medium was 0% from the tick pools and 58% from the single tick specimens.


Tumor cell-intrinsic EPHA2 suppresses anti-tumor immunity by regulating PTGS2 (COX-2).
Resistance to immunotherapy is one of the biggest problems of current oncotherapeutics. WhileT cell abundance is essential for tumor responsiveness to immunotherapy, factors that define the T cell inflamed tumor microenvironment are not fully understood. We conducted an unbiased approach to identify tumor-intrinsic mechanisms shaping the immune tumor microenvironment(TME), focusing on pancreatic adenocarcinoma because it is refractory to immunotherapy and excludes T cells from the TME. From human tumors, we identified EPHA2 as a candidate tumor intrinsic driver of immunosuppression. Epha2 deletion reversed T cell exclusion and sensitized tumors to immunotherapy. We found that PTGS2, the gene encoding cyclooxygenase-2, lies downstream of EPHA2 signaling through TGFβ and is associated with poor patient survival. Ptgs2 deletion reversed T cell exclusion and sensitized tumors to immunotherapy; pharmacological inhibition of PTGS2 was similarly effective. Thus, EPHA2-PTGS2 signaling in tumor cells regulates tumor immune phenotypes; blockade may represent a novel therapeutic avenue for immunotherapy-refractory cancers. Our findings warrant clinical trials testing the effectiveness of therapies combining EPHA2-TGFβ-PTGS2 pathway inhibitors with anti-tumor immunotherapy, and may change the treatment of notoriously therapy-resistant pancreatic adenocarcinoma.


Profiles of Language Brokering Experiences and Contextual Stressors: Implications for Adolescent Outcomes in Mexican Immigrant Families.
Adolescents from Mexican immigrant families are often embedded in a challenging social environment and experience multiple contextual stressors, including economic stress, discrimination, and foreigner stress. We consider how the effects of these contextual stressors may be amplified or diminished for adolescents who function as language brokers, interpreting and mediating for their English-limited parents. Using two waves of survey data collected from a sample (N = 604 at Wave 1; N = 483 at Wave 2) of Mexican American adolescents with ages ranging from 11 to 15 (Mage = 12.41, 54% female), four distinct brokering-stress profiles were identified. Latent profile analyses revealed that with moderate levels of contextual stress, adolescents with more positive language brokering experiences (protective group) demonstrated more favorable outcomes than those with neutral language brokering experiences (moderate group) and those who did not involve themselves as frequently in language brokering activities (less-involved group). In contrast, high levels of contextual stress, coupled with more negative language brokering experiences (risk group), produced the least favorable outcomes among adolescents.


Nutritional strategy for adolescents undergoing bariatric surgery: report of a working group of the Nutrition Committee of NASPGHAN/NACHRI.
Surgical options for the treatment of adolescent obesity have been gaining popularity. Adolescent patients present a particular challenge to clinicians, secondary to age-related issues, revolving around both mental and physical growth. These age-related issues require a unique approach to nutritional intervention for adolescents undergoing bariatric surgery as opposed to standardized approaches for adults. Despite the increasing numbers of adolescents undergoing obesity surgery, evidence-based nutritional guidelines have yet to be published. The goal of this document is to provide the clinician with recommendations on how to assess, educate, nourish, and monitor the adolescent who has undergone obesity surgery. A multidisciplinary panel composed of 3 pediatric gastroenterologists, 1 psychologist, and 3 registered dietitians from the Nutrition Committee for the North American Society of Pediatric Gastroenterology, Hepatology, and Nutrition and National Association of Children's Hospitals and Related Institutions, with experience in nutrition and adolescent weight loss surgery, reviewed the medical literature for evidence-based practice for nutritional strategies for patients undergoing bariatric surgery. In addition to this group, an adolescent medicine physician was consulted for matters related to reproductive health. The present article presents a consensus of recommendations based on a review of the literature. In areas for which there was a lack of evidence to support the recommendations, best-practice guidelines were used. The present article provides the clinician with an overview of the nutritional concerns for adolescent patients undergoing obesity surgery. These guidelines address the preoperative educational pathway, the postoperative diet progression, recognition of disordered eating, guidelines for female reproductive issues, and assistance for the adolescent in a school/college environment.


A phase I study of recombinant human interferon-alpha 2a or human lymphoblastoid interferon-alpha n1 and concomitant zidovudine in patients with AIDS-related Kaposi's sarcoma.
To determine the safety, maximum tolerated dose, and preliminary efficacy of concomitant interferon-alpha and zidovudine therapy in AIDS-related Kaposi's sarcoma (KS), 56 patients with biopsy-proven KS and documented human immunodeficiency virus type 1 (HIV) infection were enrolled into a phase I study. Interferon-alpha was given intramuscularly at a dose of 9, 18, or 27 mu once a day and zidovudine was administered as 100 or 200 mg every 4 h for 8 weeks followed by a 48-week maintenance period. The major toxicities were anemia, neutropenia, and hepatotoxicity. Neutropenia was dose limiting with 1,200 mg of zidovudine/day and the lowest dose of interferon-alpha (9 mu/day). Hepatotoxicity was dose limiting with 27 mu of interferon and 600 mg of zidovudine/day. Cumulative dose-related anemia or neutropenia was not seen during long-term follow-up. The maximum tolerated doses for the combination were defined as 18 mu daily for interferon-alpha and 600 mg daily for zidovudine. Variable changes in CD4 lymphocytes occurred during the first 8 weeks of therapy. At higher doses of the combination, sustained increases in median CD4 lymphocyte numbers were noted (p less than 0.001). In HIV antigenemic patients, progressive antigen suppression was seen with increasing doses of the combination (p less than 0.005). The overall antitumor response rate was 47%. Tumor regression was associated with better survival benefits (p less than 0.001) and a pretreatment CD4 cell count greater than or equal to 200 cells/mm3 (p = 0.01). In conclusion, intermediate doses of interferon-alpha and lower doses of zidovudine appear to be relatively well tolerated and associated with disease improvement, including survival benefits.


p53 nuclear protein overexpression in colorectal cancer: a dominant predictor of survival in patients with advanced hepatic metastases.
To determine whether p53 protein expression is similar within primary colorectal cancer (CRC) and synchronous regional and distant metastases and to assess whether p53 nuclear protein expression could predict outcome in patients with synchronous unresectable liver metastases treated by hepatic artery infusional (HAI) chemotherapy. Paraffin sections from tumor and corresponding normal mucosa representative of 50 consecutive advanced CRC cases were examined for p53 nuclear protein expression by immunohistochemistry using the monoclonal antibody PAb 1801. Patterns of p53 nuclear expression were correlated with standard clinicopathologic variables and outcome, including response to HAI and survival. In a subset analysis, the pattern of nuclear p53 immunoreactivity was compared between primary CRC and lymph node and liver metastases. Positive nuclear immunoreactivity for p53 protein was found in 72% of cases. The pattern of p53 protein expression in lymph node and liver metastases was identical to that of the primary tumor. The median survival time was 21.0 months in patients with p53-positive tumors and 53.2 months in patients with p53-negative tumors (Wilcoxon test P = .038). Two-year survival rates were 41.7% and 78.6%, respectively (P < .01). No significant difference was found in the response rates to HAI chemotherapy between the two groups. By multivariate analysis, p53 protein status was the single best predictor of survival, with a relative risk of 6.312. Our results indicate that nuclear p53 protein status in primary CRC is similar to that in metastatic sites and may be the dominant predictor of survival in patients with advanced hepatic metastases.


Benznidazole and nifurtimox nitroreductase activity in liver microsomes from male rats preinduced with phenobarbital or 3-methylcholanthrene.
Liver microsomes from Sprague-Dawley male rats are able to biotransform Benznidazole (Bz) or Nifurtimox (NFX) by nitroreduction. Pretreatment of the rats during three days with phenobarbital (80 mg/kg/day, ip) but not with 3-methylcholanthrene (35 mg/kg/day ip) increased both Bz and NFX nitroreductase activity. Results suggest that cytochrome P-450 but not cytochrome P-448 is involved in the nitroreduction of these two chemotherapeutic agents against Chagas' disease. Possible pharmacological and toxicological implications of these observations are discussed.


Avoiding fights on social media: Strategies youth leverage to navigate conflict in a digital era.
Emerging qualitative work documents that social media conflict sometimes results in violence in impoverished urban neighborhoods. Not all experiences of social media conflict lead to violence, however, and youth ostensibly use a variety of techniques to avoid violent outcomes. Little research has explored the daily violence prevention strategies youth use on social media, an important gap given the omnipresence of social media in youth culture. This paper examines youth strategies and factors that avoid violence resulting from social media conflict. Four focus groups with 41 teenagers of color solicited strategies to prevent violence resulting from social media conflict. Three coders analyzed data in Dedoose, guided by systematic textual coding using a multi-step thematic analysis. Four approaches emerged to avoiding violence from social media conflict: avoid, de-escalate, reach out for help, and bystander intervention. Our findings position youth as key players in efforts to prevent violence from resulting from social media conflict.


Factors associated with grief and depression following the loss of a child: a multivariate analysis.
The present study aims to explore the factors which are associated with grief and depression outcomes in a group of bereaved parents in the first few years following the loss of a child. Sixty-four participants were recruited from bereavement support organisations, between two and 59 months post loss, mean 30 months (SD = 15). They completed a questionnaire packet which comprised standard instruments measuring grief, depression, coping styles, continuing bonds and optimism/pessimism, as well as a number of specific bereavement-related questions. Univariate analyses were conducted to establish which factors were associated with grief and depression. Those which were statistically significant were then entered into multivariate analyses to establish their relative importance. High levels of avoidance and depression and lower levels of cognitive restructuring (benefit finding) were associated with higher grief symptoms, whereas higher levels of avoidance and alcohol/substance use were associated with higher depression symptoms. The present study highlights the relative importance of different coping strategies adopted by this group of bereaved parents, compared to the relative unimportance of circumstances around the loss, e.g. sudden or violent death. The use of alcohol and other substances by bereaved parents requires urgent attention as a potentially life-threatening maladaptive coping strategy. The call for further research into risk factors for bereaved parents is emphasised.


Metaphor in psychosis: on the possible convergence of Lacanian theory and neuro-scientific research.
Starting from the theories of leading psychiatrists, like Kraepelin and de Clérambault, the French psychoanalyst Jacques Lacan (1901-1981) formulated an original theory of psychosis, focusing on the subject and on the structuring role of language. In particular, he postulated that language makes up the experience of subjectivity and that psychosis is marked by the absence of a crucial metaphorization process. Interestingly, in contemporary psychiatry there is growing empirical evidence that schizophrenia is characterized by abnormal interpretation of verbal and non-verbal information, with a great difficulty to put such information in the appropriate context. Neuro-scientific contributions have investigated this difficulty suggesting the possibility of interpreting schizophrenia as a semiotic disorder which makes the patients incapable of understanding the figurative meaning of the metaphoric speech, probably due to a dysfunction of certain right hemisphere areas, such as the right temporoparietal junction and the right superior/middle temporal gyrus. In this paper we first review the Lacanian theory of psychosis and neuro-scientific research in the field of symbolization and metaphoric speech. Next, we discuss possible convergences between both approaches, exploring how they might join and inspire one another. Clinical and neurophysiological research implications are discussed.


Extracorporeal perfusion - reduced to a one-way infusion.
Extracorporeal perfusion (EP) is moving into focus of research in reconstructive and transplantation medicine for the preservation of amputates and free tissue transplants. The idea behind EP is the reduction of ischemia-related cell damage between separation from blood circulation and reanastomosis of the transplant. Most experimental approaches are based on a complex system that moves the perfusate in a circular course. In this study, we aimed to evaluate if a simple perfusion by an infusion bag filled with an electrolyte solution can provide acceptable results in terms of flow stability, oxygen supply and viability conservation for EP of a muscle transplant. The results are compared to muscles perfused with a pump system as well as muscles stored under ischemic conditions after a one-time intravasal flushing with Jonosteril. With this simple method a sufficient oxygen supply could be achieved and functionality could be maintained between 3.35 times and 4.60 times longer compared to the control group. Annexin V positive nuclei, indicating apoptosis, increased by 9.7% in the perfused group compared to 24.4% in the control group. Overall, by decreasing the complexity of the system, EP by one-way infusion can become more feasible in clinical situations.


Anti-(insulin receptor) monoclonal antibody-stimulated tyrosine phosphorylation in cells transfected with human insulin receptor cDNA.
The effects of insulin and anti-(insulin receptor) monoclonal antibodies on tyrosine phosphorylation were investigated in fibroblasts transfected with human insulin receptor cDNA (NIH 3T3HIR3.5 cells) using anti-phosphotyrosine immunoblotting. Insulin increased levels of tyrosine phosphorylation in two major proteins of molecular mass 97 kDa (pp97, assumed to be the insulin receptor beta-subunit) and 185 kDa (pp185). Insulin-mimetic anti-receptor antibodies also stimulated tyrosine phosphorylation of both pp97 and pp185. The observation of antibody-stimulated pp97 phosphorylation, as detected by immunoblotting, is in contrast with previous data which failed to show receptor autophosphorylation in NIH 3T3HIR3.5 cells labelled with [32P]P1. The effect of insulin on pp97 was maximal within 1 min, but the response to antibody was apparent only after a lag of 1-2 min and rose steadily over 20 min. The absolute level of antibody-stimulated phosphorylation of both pp97 and pp185 after 20 min was only about 20% of the maximum level induced by equivalent concentrations of insulin, even at concentrations of antibody sufficient for full occupancy of receptors. Another insulin-mimetic agent, wheat-germ agglutinin, stimulated receptor autophosphorylation with kinetics similar to those produced by the antibody. It is suggested that the relatively slow responses to both agents may be a function of the dependence on receptor cross-linking. These data are consistent with a role for the insulin receptor tyrosine kinase activity in the mechanism of action of insulin-mimetic anti-receptor antibodies.


Pharmacology and signaling of MAS-related G protein-coupled receptors.
Signaling by heptahelical G protein-coupled receptors (GPCR) regulates many vital body functions. Consequently, dysfunction of GPCR signaling leads to pathologic states, and approximately 30% of all modern clinical drugs target GPCR. One decade ago, an entire new GPCR family was discovered, which was recently named MAS-related G protein-coupled receptors (MRGPR) by the HUGO Gene Nomenclature Committee. The MRGPR family consists of ∼40 members that are grouped into nine distinct subfamilies (MRGPRA to -H and -X) and are predominantly expressed in primary sensory neurons and mast cells. All members are formally still considered "orphan" by the Committee on Receptor Nomenclature and Drug Classification of the International Union of Basic and Clinical Pharmacology. However, several distinct peptides and amino acids are discussed as potential ligands, including β-alanine, angiotensin-(1-7), alamandine, GABA, cortistatin-14, and cleavage products of proenkephalin, pro-opiomelanocortin, prodynorphin, or proneuropeptide-FF-A. The full spectrum of biologic roles of all MRGPR is still ill-defined, but there is evidence pointing to a role of distinct MRGPR subtypes in nociception, pruritus, sleep, cell proliferation, circulation, and mast cell degranulation. This review article summarizes findings published in the last 10 years on the phylogenetic relationships, pharmacology, signaling, physiology, and agonist-promoted regulation of all MRGPR subfamilies. Furthermore, we highlight interactions between MRGPR and other hormonal systems, paying particular attention to receptor multimerization and morphine tolerance. Finally, we discuss the challenges the field faces presently and emphasize future directions of research.


Investigation into the functionality of controlled drug denaturing/destruction kits.
Throughout the UK a large amount of unwanted, expired or patient returned controlled drugs are disposed of every day, in community and hospital pharmacies, veterinary surgeries, hospices, private hospitals and industrial settings. This is mostly achieved through the use of commercially available controlled drug destruction/denaturing kits, but what do these kits actually do to the drug within them? The primary aim of this study was to investigate the effect of six commercially available kits on morphine, a chosen model controlled drug. The secondary aim was to establish if the kits could be adapted to chemically destroy any drug disposed within it. Morphine was dispensed in to six commercially available controlled drug destruction kits at a known concentration. The instructions on the kits were followed and after 48 h the amount of drug remaining was determined by HPLC. In addition a new kit containing sodium perborate was tested in the same way. Between 78 and 111% of the parent drug was found to still be present in the commercial kits tested after 48 h. In the sodium perborate 5% kit this level fell to 22%. In conclusion all the commercially available CD denaturing kits tested do not destroy the controlled drug (morphine) tested but simply encapsulated it in gel. This means the parent form of the drug is still present and could potentially be recovered and abused. The new kit containing sodium perborate was much more effective in chemically destroying the parent drug but care must be taken in its use.


Performance of K-ras mutation analysis plus endoscopic ultrasound-guided fine-needle aspiration for differentiating diagnosis of pancreatic solid mass: a meta-analysis.
Difficulties persist in differentiating pancreatic ductal adenocarcinomas (PDAC) from pancreatic inflammatory masses (PIM). Auxiliary diagnostic techniques which enhance the endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) diagnostic yield have been attempted, for example, K-ras mutation analysis. We aimed to evaluate the accuracy of K-ras mutation analysis combined with EUS-FNA for the differential diagnosis of PDAC and PIM by pooling data of existing trials. We systematically searched the Medline, PubMed, Web of Science, Embase, and Cochrane Central Trials databases for relevant published studies. Meta-analysis was performed. Pooling was conducted in fixed-effect model or random-effect model. In total eight studies, with 696 cases of PDAC and 138 cases of PIM, met our inclusion criteria. The pooled sensitivity, specificity, positive likely ratio and negative likely ratio of K-ras mutation analysis combined with cytopathology for diagnosis of PDAC versus PIM were 90%, 95%, 13.45, and 0.13, respectively. Especially, among total 123 patients whose EUS-FNA results were inconclusive or negative, fifty-nine had K-ras mutations and were finally diagnosed with PDAC (48%, 59/123). Publication bias was not present. Combining K-ras mutation analysis with routine cytology moderately improves the ability of EUS-FNA to differentially diagnose between PDAC and PIM, especially for patients with suspected PDAC yet inconclusive EUS-FNA findings, and may prove to be a valuable supplemental method to EUS-FNA.


The Interface between the State and NGOs in Delivering Health Services in Zimbabwe-A Case of the MSF ART Programme.
An over-reliance on donor funding for HIV/AIDS healthcare services remains a concern in Africa. This study, therefore, explores the partnership between the Zimbabwean government and an international non-governmental organisation in delivering HIV/AIDS healthcare services. An interpretivist paradigm and descriptive phenomenological design were used to elicit the opinions, perceptions, and experiences of forty purposively sampled key informants. Thematic analysis was employed using ATLAS.ti version 7.1.4 to analyse the data. The differences in terms of policies, structures, and administrative issues between the partners identified challenges in the implementation of the programme. This was demonstrated through the reversal of the gains attained in prevention, care, and treatment. This raises concerns for increased risk of defaulters, drug resistance, and deaths. Therefore, the partners in this endeavour should negotiate an aligned approach for the efficient delivery of HIV/AIDS healthcare services.


Phosphorylation-dependent proteome of Marcks in ependyma during aging and behavioral homeostasis in the mouse forebrain.
Ependymal cells (ECs) line the ventricular surfaces of the mammalian central nervous system (CNS) and their development is indispensable to structural integrity and functions of the CNS. We previously reported that EC-specific genetic deletion of the myristoylated alanine-rich protein kinase C substrate (Marcks) disrupts barrier functions and elevates oxidative stress and lipid droplet accumulation in ECs causing precocious cellular aging. However, little is known regarding the mechanisms that mediate these changes in ECs. To gain insight into Marcks-mediated mechanisms, we performed mass spectrometric analyses on Marcks-associated proteins in young and aged ECs in the mouse forebrain using an integrated approach. Network analysis on annotated proteins revealed that the identified Marcks-associated complexes are in part involved in protein transport mechanisms in young ECs. In fact, we found perturbed intracellular vesicular trafficking in cultured ECs with selective deletion of Marcks (Marcks-cKO mice), or upon pharmacological alteration to phosphorylation status of Marcks. In comparison, Marcks-associated protein complexes in aged ECs appear to be involved in regulation of lipid metabolism and responses to oxidative stress. Confirming this, we found elevated signatures of inflammation in the cerebral cortices and the hippocampi of young Marcks-cKO mice. Interestingly, behavioral testing using a water maze task indicated that spatial learning and memory is diminished in young Marcks-cKO mice similar to aged wildtype mice. Taken together, our study provides first line of evidence for potential mechanisms that may mediate differential Marcks functions in young and old ECs, and their effect on forebrain homeostasis during aging.


Antibiotic therapy versus no antibiotic therapy for children aged two to 59 months with WHO-defined non-severe pneumonia and wheeze.
Worldwide, pneumonia is the leading cause of death among children under five years of age and accounts for approximately two million deaths annually. The World Health Organization (WHO) has developed case management guidelines based on simple clinical signs to help clinicians decide on the appropriate pneumonia treatment. Children and infants who exhibit fast breathing (50 breaths per minute or more in infants two months to 12 months of age and 40 or more in children 12 months to five years of age) and cough are presumed to have non-severe pneumonia and the WHO recommends antibiotics. Implementation of these guidelines to identify and manage pneumonia at the community level has been shown to reduce acute respiratory infection (ARI)-related mortality by 36%, although apprehension exists regarding these results due to the questionable quality of evidence. As WHO guidelines do not make a distinction between viral and bacterial pneumonia, these children continue to receive antibiotics because of the concern that it may not be safe to do otherwise. Therefore, it is essential to explore the role of antibiotics in children with WHO-defined non-severe pneumonia and wheeze and to develop effective guidelines for initial antibiotic treatment. To evaluate the efficacy of antibiotic therapy versus no antibiotic therapy for children aged two to 59 months with WHO-defined non-severe pneumonia and wheeze. We searched CENTRAL (2014, Issue 1), MEDLINE (1946 to March week 3, 2014), EMBASE (January 2010 to March 2014), CINAHL (1981 to March 2014), LILACS (1982 to March 2014), Networked Digital Library of Theses and Dissertations (23 July 2013) and Web of Science (1985 to March 2014). Randomised controlled trials (RCTs) evaluating the efficacy of antibiotic therapy versus no antibiotic therapy for children aged two to 59 months with non-severe pneumonia and wheeze. We considered studies that defined non-severe pneumonia as cough or difficulty in breathing with a respiratory rate above the WHO-defined age-specific values (respiratory rate of 50 breaths per minute or more for children aged two to 12 months, or a respiratory rate of 40 breaths per minute or more for children aged 12 to 59 months) and wheeze for inclusion. We have excluded non-RCTs (quasi-RCTs). Two review authors independently assessed the search results and extracted data. We did not identify any study that completely fulfilled our inclusion criteria. There is a clear need for RCTs to address this question in representative populations. We do not currently have evidence to support or challenge the continued use of antibiotics for the treatment of non-severe pneumonia, as suggested by WHO guidelines.


Hepatic Manifestations in Wilson's Disease: Report of 110 Cases.
Wilson's disease is a rare disease and difficult to establish diagnosis. We aim to improve understanding and early diagnosis. Medical records were reviewed for 110 patients with Wilson's disease. The clinical manifestations and laboratory findings were retrospectively analyzed, especially in terms of age, type of liver injury. Age range at diagnosis was wide (4 to 52 years).The most frequent hepatic manifestations observed were jaundice (40.9%), fatigue (37.3%), nausea or vomiting (32.7%) and bloating (30.0%). Hepatic involvement in affected patients may take one of several different presentations. Thirty-eight patients were found cirrhosis with asymptomatic or slowly progressive hepatic dysfunction. Twelve were acute liver failure superimposed on chronic cirrhosis. Fifteen were acute hepatic failure without cirrhosis. Nineteen presented as acute hepatitis. Four showed chronic liver dysfunction. Five were asymptomatic aminotransferasemia. Another 17 patients showed neurological disorders with cirrhosis. Kayser-Fleischer rings were found in 91.3% patients. The serum ceruloplasmin decreased in 85.1%, 24-hour urinary copper increased in 83.9%, and serum copper decreased in 61.9% patients. The clinical manifestation of Wilson's disease is very diverse and no one feature is completely reliable. Patients at any age with liver injury of unknown etiology should be screened for Wilson's disease.


Classification of inattentional blindness using brain dynamics of ERPs.
Situational awareness (SA) is vital for understanding our surroundings. Multiple variables, including inattentive blindness (IB), contribute to the deterioration of SA, which may have detrimental effects on individuals' cognitive performance. IB occurs due to attentional limitations, ignoring critical information and resulting in a loss of SA and a decline in general performance, particularly in complicated situations requiring substantial cognitive resources. To the best of our knowledge, however, past research has not fully uncovered the neurological characteristics of IB nor classified these characteristics in life-alike virtual situations. Therefore, the purpose of this study is to determine whether ERP dynamics in the brain may be utilised as a neural feature to predict the occurrence of IB using machine learning (ML) algorithms. In a virtual reality simulation of an IB experiment, 30 participants' behaviour and Electroencephalography (EEG) measurements were obtained. Participants were given a target detection task in the IB experiment without knowing the unattended shapes displayed on the background building. The targets were presented in three different sensory modalities (auditory, visual, and visual-auditory). On the post-experiment questionnaire, participants who claimed not to have noticed the unattended shapes were assigned to the IB group. Subsequently, the Aware group was formed from individuals who reported seeing the unattended shapes. Using EEGNet to classify IB and Aware groups demonstrated a high classification performance. According to the research, ERP brain dynamics are associated with the awareness of unattended shapes and have the potential to serve as a reliable indication for predicting the visual consciousness of unexpected objects.(p/)(p)Clinical relevance- This research offers a potential brain marker for the mixed-reality and BCI systems that will be used in the future to identify cognitive deterioration, maintain attentional capacity, and prevent disasters.


Differential anti-inflammatory effects of immunosuppressive drugs: cyclosporin, rapamycin and FK-506 on inducible nitric oxide synthase, nitric oxide, cyclooxygenase-2 and PGE2 production.
Cyclosporin, FK-506 and rapamycin have similar but distinct modes of interaction with cyclophilins, calcineurins and transcription factors. These immunosuppressive drugs have also been shown to inhibit cytotoxic and inflammatory responses in macrophage. Therefore, we evaluated the mechanism of action of these drugs on iNOS and COX-2 expression by macrophages, the products of which (NO and PGE2) have cytotoxic and proinflammatory activities. The murine macrophage cell line RAW 264.7 was grown as monolayer cultures. The effects of pharmacologically relevant concentrations of cyclosporin, rapamycin and FK-506 were evaluated in the presence and absence of lipopolysaccharide (LPS) which is a known inducer of iNOS and COX-2. Subsequently the expression of iNOS and COX-2 were analyzed by Western and Northern analysis. The production of NO and PGE2 were assayed by Greiss and RIA respectively. Cyclosporin (1-5 microg/ml) and rapamycin (1.0-10 nM) but not FK-506 (5-10 nM) inhibited both iNOS and COX-2 expression at mRNA level which led to significant inhibition of NO and PGE2 production. These studies characterize differential mechanistic capacity of the immunophilin-binding immunosuppressive drugs (comparable to hydrocortisone) to inhibit both iNOS and COX-2 expression. Inhibition of iNOS and COX-2 mRNA accumulation by cyclosporin and rapamycin seem to be distinct. These studies also highlight potential anti-inflammatory properties of these drugs in addition to their known immunosuppressive activity.


Successful treatment of refractory juvenile generalized pustular psoriasis with secukinumab monotherapy: A case report and review of published work.
Juvenile generalized pustular psoriasis (GPP) is rare and often resistant to conventional systemic therapy such as methotrexate, retinoic acid and cyclosporin A. GPP can be induced by deficiency of interleukin (IL)-36 receptor antagonist (DITRA). No standardized guidelines are available for juvenile GPP or DITRA, and a uniformly safe and effective biologic agent has not been identified. However, multiple biologics approved for use in plaque-type psoriasis have also been used in GPP. Herein, we report a case of a 6-year-old Taiwanese boy with GPP and homozygous mutation at c.115+6T>C within the IL-36 receptor antagonist (IL36RN) gene, who was treated successfully with secukinumab after failure of prior methotrexate, acitretin, cyclosporin A, etanercept and adalimumab. Similar to two previously reported non-adult cases of GPP successfully treated with secukinumab, our case also demonstrated a history of repeated treatment failures with several conventional oral systemic agents and biologics. Different from these two cases, however, ours is the first of juvenile GPP successfully treated with secukinumab monotherapy, without using other systemic agent concurrently during the use of secukinumab.


Empathy and Language in the Clinician Patient Relationship: Improving the Translation of Evidence to Practice.
For clinicians, effective communication goes beyond the delivery of scientific information to include an understanding of who the patient is and what they value; treating them with respect; and acknowledging their emotional and social realities. Recognizing our own implicit biases and cultivating a more mindful approach to the impact of language, especially around weight and food, and using verbal and nonverbal approaches to convey empathic concern can improve our relationships with our patients. Patients who feel seen and respected are more likely to follow through with recommendations including dietary change, leading to improved health and quality of life.


Osteointegrative and microgeometric comparison between micro-blasted and alumina blasting/acid etching on grade II and V titanium alloys (Ti-6Al-4V).
This study evaluated the effect of alumina-blasted/acid-etched (AB/AE) or microabrasive blasting (C3-Microblasted) surface treatment on the osseointegration of commercially-pure Ti (grade II) and Ti-6Al-4V alloy (grade V) implants compared to as-machined surfaces. Surface characterization was performed by scanning electron microscopy and optical interferometry (IFM) to determine roughness parameters (Sa and Sq, n = 3 per group). One-hundred forty-four implants were placed in the radii of 12 beagle dogs, for histological (n = 72, bone-to-implant contact - BIC and bone-area-fraction occupancy -BAFO) and torque to interface failure test at 3 and 6 weeks (n = 72). SEM and IFM revealed a significant increase in surface texture for AB/AE and C3-Microblasted surfaces compared to machined surface, regardless of titanium substrate. Torque-to-interface failure test showed significant increase in values from as-machined to AB/AE and to C3-Microblasted. Considering time in vivo, alloy grade, and surface treatment, the C3-microblasted presented higher mean BIC values relative to AB/AE and machined surfaces for both alloy types. BAFO levels were significantly higher for both textured surfaces groups relative to the machined group at 3 weeks, but differences were not significant between the three surfaces for each alloy type at 6 weeks. Surface treatment resulted in roughness that improved osseointegration in Grade II and V titanium substrates.


TCA Cycle and Mitochondrial Membrane Potential Are Necessary for Diverse Biological Functions.
Mitochondrial metabolism is necessary for the maintenance of oxidative TCA cycle function and mitochondrial membrane potential. Previous attempts to decipher whether mitochondria are necessary for biological outcomes have been hampered by genetic and pharmacologic methods that simultaneously disrupt multiple functions linked to mitochondrial metabolism. Here, we report that inducible depletion of mitochondrial DNA (ρ(ο) cells) diminished respiration, oxidative TCA cycle function, and the mitochondrial membrane potential, resulting in diminished cell proliferation, hypoxic activation of HIF-1, and specific histone acetylation marks. Genetic reconstitution only of the oxidative TCA cycle function specifically in these inducible ρ(ο) cells restored metabolites, resulting in re-establishment of histone acetylation. In contrast, genetic reconstitution of the mitochondrial membrane potential restored ROS, which were necessary for hypoxic activation of HIF-1 and cell proliferation. These results indicate that distinct mitochondrial functions associated with respiration are necessary for cell proliferation, epigenetics, and HIF-1 activation.


Recent advances in the solid phase synthesis of drug-like heterocyclic small molecules.
Phage display is a biological system which facilitates the cloning and rapid selection of peptides from large combinatorial libraries. In compa-rison to the chemical combinatorial approach, the advantages of phage display lie in its simplicity and replicability. While phage display has many diverse applications, this review will focus on the use of phage peptide libraries to discover epitopes recognised by monoclonal antibodies. As monoclonal antibodies are useful tools for the detection of proteins and for the investigation of molecular interactions, the identification of their epitopes will serve to elucidate the structure and function of proteins, as well as aid in the discovery of new drugs and the development of vaccines.


A comparison of the effect of intranasal desmopressin and intramuscular hyoscine N-butyl bromide combination with intramuscular hyoscine N-butyl bromide alone in acute renal colic.
Patients with acute renal colic usually require immediate diagnosis and treatment. In this clinical trial analgesic effect of hyoscine N-butyl bromide and desmopressin combination in comparison with hyoscine N-butyl bromide alone in patients with acute renal colic induced by urinary stones was assessed. The study included 114 patients randomly allocated in two groups (A and B). Patients in group A received 20 mg intramuscular hyoscine N-butyl bromide at admission time and patients in group B received 20 μg of intranasal desmopressin in combination with 20 mg intramuscular hyoscine N-butyl bromide. A visual analogue scale (VAS; a 10-cm horizontal scale ranging from "zero or no pain" to "10 or unbearable pain") was hired to assess the patients' pain severity at baseline, 30 and 60 minutes after the treatments. ON ADMISSION, THE PAIN LEVEL WAS SIMILAR IN BOTH GROUPS (GROUP A: 8.95 ± 0.11 and group B: 8.95 ± 0.12). In group A, the mean of pain level showed a decrease after 30 minutes (group A: 7.26 ± 0.25 and group B: 5.95 ± 0.28) but further decreasing did not occur; however in group B, the pain consistently decreased and the mean after 60 minutes was significantly decreased (group A: 6.80 ± 0.31 and group B: 3.71 ± 0.31). No side effects were detected in this study. The combination of hyoscine N-butyl bromide and desmopressin is more effective than hyoscine N-butyl bromide alone in patients with renal colic. Further studies are recommended to validate these findings and compare the different doses of desmopressin.


Staphylococcus carnosus: from starter culture to protein engineering platform.
Since the 1950s, Staphylococcus carnosus is used as a starter culture for sausage fermentation where it contributes to food safety, flavor, and a controlled fermentation process. The long experience with S. carnosus has shown that it is a harmless and "food grade" species. This was confirmed by the genome sequence of S. carnosus TM300 that lacks genes involved in pathogenicity. Since the development of a cloning system in TM300, numerous genes have been cloned, expressed, and characterized and in particular, virulence genes that could be functionally validated in this non-pathogenic strain. A secretion system was developed for production and secretion of industrially important proteins and later modified to also enable display of heterologous proteins on the surface. The display system has been employed for various purposes, such as development of live bacterial delivery vehicles as well as microbial biocatalysts or bioadsorbents for potential environmental or biosensor applications. Recently, this surface display system has been utilized for display of peptide and protein libraries for profiling of protease substrates and for generation of various affinity proteins, e.g., Affibody molecules and scFv antibodies. In addition, by display of fragmented antigen-encoding genes, the surface expression system has been successfully used for epitope mapping of antibodies. Reviews on specific applications of S. carnosus have been published earlier, but here we provide a more extensive overview, covering a broad range of areas from food fermentation to sophisticated methods for protein-based drug discovery, which are all based on S. carnosus.


A multidisciplinary approach to enhance implant esthetics: case report.
Esthetic implant-supported restorations in the anterior maxilla present a number of clinical challenges to the dentist, including the need to preserve or restore the periimplant hard and soft tissue architecture around the implant site. In this clinical case report, two implant systems were used to restore two missing maxillary central incisors to make use of the physical properties in each system. Two different surgical protocols were used to place each implant, and the deficient alveolar bone was restored with an autogenous bone graft. After a detailed clinical evaluation, a comprehensive treatment plan was developed to address the unique demands of the patient. Both teeth were fully restored with esthetic, implant-supported, single-tooth restorations, and clinical results were recorded postoperatively.


Nanosolids, slushes, and nanoliquids: characterization of nanophases in metal clusters and nanoparticles.
One of the keys to understanding the emergent behavior of complex materials and nanoparticles is understanding their phases. Understanding the phases of nanomaterials involves new concepts not present in bulk materials; for example, the phases of nanoparticles are quantum mechanical even when no hydrogen or helium is present. To understand these phases better, molecular dynamics (MD) simulations on size-selected particles employing a realistic analytic many-body potential based on quantum mechanical nanoparticle calculations have been performed to study the temperature-dependent properties and melting transitions of free Al n clusters and nanoparticles with n = 10-300 from 200 to 1700 K. By analyzing properties of the particles such as specific heat capacity (c), radius of gyration, volume, coefficient of thermal expansion (beta), and isothermal compressibility (kappa), we developed operational definitions of the solid, slush, and liquid states of metal clusters and nanoparticles. Applying the definitions, which are based on the temperature dependences of c, beta, and ln kappa, we determined the temperature domains of the solid, slush, and liquid states of the Al n particles. The results show that Al n clusters ( n <or= 18, diameter of less than 1 nm) are more like molecules, and it is more appropriate to say that they have no melting transition, but Al n nanoparticles ( n >or= 19, diameter of more than 1 nm) do have a melting transition and are in the liquid state above 900-1000 K. However, all aluminum nanoparticles have a wide temperature interval corresponding to the slush state in which the solid and liquid states coexist in equilibrium, unlike a bulk material where coexistence occurs only at a single temperature (for a given pressure). The commonly accepted operational marker of the melting temperature, namely, the peak position of c, is not unambiguous and not appropriate for characterizing the melting transition for aluminum particles with the exception of a few particle sizes that have a single sharp peak (as a function of temperature) in each of the three properties, c, beta, and ln kappa.


Incentivising research data sharing: a scoping review.
Background: Numerous mechanisms exist to incentivise researchers to share their data. This scoping review aims to identify and summarise evidence of the efficacy of different interventions to promote open data practices and provide an overview of current research. Methods: This scoping review is based on data identified from Web of Science and LISTA, limited from 2016 to 2021. A total of 1128 papers were screened, with 38 items being included. Items were selected if they focused on designing or evaluating an intervention or presenting an initiative to incentivise sharing. Items comprised a mixture of research papers, opinion pieces and descriptive articles. Results: Seven major themes in the literature were identified: publisher/journal data sharing policies, metrics, software solutions, research data sharing agreements in general, open science 'badges', funder mandates, and initiatives. Conclusions: A number of key messages for data sharing include: the need to build on existing cultures and practices, meeting people where they are and tailoring interventions to support them; the importance of publicising and explaining the policy/service widely; the need to have disciplinary data champions to model good practice and drive cultural change; the requirement to resource interventions properly; and the imperative to provide robust technical infrastructure and protocols, such as labelling of data sets, use of DOIs, data standards and use of data repositories.


Pancreatic islet discrimination of hexose anomers. II. Transient computer simulation.
We have previously modeled pancreatic islet glycolysis under idealized steady-state conditions where the input is a pure hexose anomer and there is no mutarotation and reproduced the known preference for the alpha-anomers of glucose and mannose as substrates. This model is here extended to simulate real experiments, where the hexoses mutarotate and measurements may be taken over time. The behavior of our model system agrees with available experimental data. The hexose diphosphate activators of phosphofructokinase, whose effect was seen as not important in the preceding steady-state analysis, are found here to have a modest (approximately 10-15%) effect on its flux. The previous conclusion that the anomeric preference of the glycolytic pathway follows from that of glucokinase continues to hold in the real experimental situation.


Paraneoplastic remitting seronegative symmetrical synovitis with pitting edema (RS3PE syndrome): a report of two cases and review of the literature.
We report two cases of paraneoplastic remitting seronegative symmetrical synovitis with pitting edema (RS3PE) associated with prostatic adenocarcinoma. One of the patients was positive for Helicobacter pylori and the other had secondary bone metastases. In the latter, the clinical picture of RS3PE developed after surgical intervention for the primary lesion. On physical examination, while the hands and feet were swollen in the first patient, pitting edema was present only at the feet of the other. All joints of the affected hands and feet were painful. Serological tests including rheumatoid factor, antinuclear antibody, and human leukocyte antigen B27 were all negative. Response to low-dose corticosteroid treatment was delayed in the first patient, but the symptoms were relieved better in the second one.


The use of simulation in emergency medicine: a research agenda.
Medical simulation is a rapidly expanding area within medical education. In 2005, the Society for Academic Emergency Medicine Simulation Task Force was created to ensure that the Society and its members had adequate access to information and resources regarding this new and important topic. One of the objectives of the task force was to create a research agenda for the use of simulation in emergency medical education. The authors present here the consensus document from the task force regarding suggested areas for research. These include opportunities to study reflective experiential learning, behavioral and team training, procedural simulation, computer screen-based simulation, the use of simulation for evaluation and testing, and special topics in emergency medicine. The challenges of research in the field of simulation are discussed, including the impact of simulation on patient safety. Outcomes-based research and multicenter efforts will serve to advance simulation techniques and encourage their adoption.


[Advanced lung cancer with recurrence of liver and tracheal metastases responsive to multimodality therapy - a case report].
We report a case of advanced lung cancer with recurrence of liver and tracheal metastases that were responsive to multimodality therapy. The patient was a 77-year-old man who suffered from advanced lung cancer with chronic obstructive pulmonary disease (COPD) and alcohol-induced liver cirrhosis. The primary lung cancer was surgically resected. Eight months after resection of the primary lung cancer, a solitary liver tumor appeared and hepatic resection was performed. Histological findings showed that both the primary lung tumor and the solitary liver tumor were squamous cell carcinoma (SCC). Subsequently, he developed a recurrence in his trachea 8 months after hepatic resection. Radiotherapy, endobronchial argon plasma coagulation (APC), and systemic chemotherapy were administered. The tracheal tumor remained stable without any liver metastasis for 25 months.


Norepinephrine increases Na+-K+-ATPase and solute transport in rabbit proximal tubules.
We studied the effects of norepinephrine on solute transport and oxidative metabolism in proximal tubules. Norepinephrine (10(-6) M) in the bath stimulated fluid absorption (Jv) by proximal convoluted tubules from 0.76 +/- 0.10 to 1.01 +/- 0.11 nl X mm-1 X min-1 (P less than 0.001). Bicarbonate, chloride, and phosphate transport also increased in proportion to the increases in Jv. Norepinephrine increased ouabain-sensitive oxygen consumption (QO2) in suspensions of cortical tubules by 1.3 nmol X mg protein-1 X min-1 and had no effect on ouabain-insensitive QO2 or mitochondrial respiration. Na+-K+-ATPase activity in basolateral membranes prepared from cortical homogenates incubated with norepinephrine increased from 277.1 +/- 34.9 to 411.1 +/- 38.6 nmol Pi X mg protein-1 X min-1 (P less than 0.005). Norepinephrine also increased Na+-K+-ATPase activity of cortical homogenates by 72% but had no effect on Na+-K+-ATPase if added directly to purified basolateral membranes. These studies show that norepinephrine stimulates solute transport in the proximal tubule by increasing Na+-K+-ATPase activity indirectly through some component or components of the adrenergic receptor system.


Long-term dietary quality and risk of developing rheumatoid arthritis in women.
To evaluate the association between long-term dietary quality, measured by the 2010 Alternative Healthy Eating Index, and risk of rheumatoid arthritis (RA) in women. We prospectively followed 76 597 women in the Nurses' Health Study aged 30-55 years and 93 392 women in the Nurses' Health Study II aged 25-42 years at baseline and free from RA or other connective tissue diseases. The lifestyle, environmental exposure and anthropometric information were collected at baseline and updated biennially. Cumulative follow-up rates were more than 90% for both cohorts. The primary outcome was RA alone with two subtypes of the disease: seropositive and seronegative RA. During 3 678 104 person-years, 1007 RA cases were confirmed. In the multivariable-adjusted model, long-term adherence to healthy eating patterns was marginally associated with reduced RA risk. To assess potential effect modification by age at diagnosis, we stratified by age. Among women aged ≤55 years, better quality diet was associated with lower RA risk (HRQ4 vs Q1: 0.67; 95% CI 0.51 to 0.88; p trend: 0.002), but no significant association was found for women aged >55 years (p interaction: 0.005). When stratifying by serostatus, the inverse association among those aged ≤55 years was strongest for seropositive RA (HRQ4 vs Q1: 0.60; 95% CI 0.42 to 0.86; p trend: 0.003). A healthier diet was associated with a reduced risk of RA occurring at 55 years of age or younger, particularly seropositive RA.


Concise and Stereodivergent Synthesis of Carbasugars Reveals Unexpected Structure-Activity Relationship (SAR) of SGLT2 Inhibition.
Carbasugar sodium-glucose cotransporter 2 (SGLT2) inhibitors are highly promising drug candidates for the treatment of Type 2 diabetes mellitus (T2DM). However, the clinical usage of carbasugar SGLT2 inhibitors has been underexplored, due to the lengthy synthetic routes and the lack of structure-activity relationship (SAR) studies of these compounds. Herein, we report a concise and stereodivergent synthetic route towards some novel carbasugar SGLT2 inhibitors, featuring an underexploited, regioselective, and stereospecific palladium-catalyzed allyl-aryl coupling reaction. This synthetic strategy, together with computational modeling, revealed the unexpected SAR of these carbasugar SGLT2 inhibitors, and enabled the discovery of a highly selective and potent SGLT2 inhibitor.


Functionality of intrinsic disorder in tumor necrosis factor-α and its receptors.
Tumor necrosis factor-α (TNF-α) is a pleiotropic inflammatory cytokine that exerts potent cytotoxic effects on solid tumor cells, while not affecting their normal counterparts. It is also known that TNF-α exerts many of its biological functions via interaction with specific receptors. To understand the potential roles of intrinsic disorder in the functioning of this important cytokine, we explored the peculiarities of intrinsic disorder distribution in human TNF-α and its homologs from various species, ranging from zebrafish to chimpanzee. We also studied the peculiarities of intrinsic disorder distribution in human TNF-α receptors, TNFR1 and TNFR2. Analysis revealed that cytoplasmic domains of TNF-α and its receptors are expected to be highly disordered. Furthermore, although the sequence identities of analyzed TNF-α homologs range from 99.57% (between human and chimpanzee proteins) to 22.33% (between frog and fish proteins), their intrinsic disorder profiles are characterized by a remarkable similarity. These observations indicate that the peculiarities of distribution of the intrinsic disorder propensity within the amino acid sequences are evolutionary conserved, and therefore could be of functional importance for this family of proteins. We also show that disordered and flexible regions of human TNF-α and its TNFR1 and TNFR2 receptors are crucial for some of their biological activities.