Clinical significance of ostial great saphenous vein reflux.
To evaluate anatomical and haemodynamic differences in patients with great saphenous vein (GSV) insufficiency by duplex scanning and air plethysmography. Duplex scanning and air plethysmography examination were undertaken. One hundred and twenty-one limbs in 91 patients were selected prospectively and divided into three groups: group A consisted of 27 controls; group B consisted of 25 limbs with GSV reflux and normal saphenous femoral junction (SFJ) and group C consisted of 69 limbs of patients with GSV and SFJ reflux. The presence of reflux and GSV diameter (SFJ, proximal and medial thirds of the thigh, the knee and medial and distal thirds of the calf) were assessed by duplex scanning. Air plethysmography was used to evaluate haemodynamic parameters: total venous volume (VV), venous filling index (VFI), residual volume fraction (RVF) and ejection fraction (EF). There was a significant difference in GSV diameter among the three groups in almost all segments evaluated (e.g. medial thigh group A = 2.4 SD 0.3 mm; B = 3.2 SD 0.7 mm; C = 5.9 SD 2.2mm p<0.001, Anova). A significant difference in VFI was found among the groups (group A = 1.2 SD 0.5; B = 2.0 SD 1.4; C = 4.0 SD 2.5 p<0.05, Anova). VV was statistical different between groups A and C (p = 0.004) and B and C(p = 0.03). EF and RVF were comparable in all groups. The VFI was normal in 68% in group B comparing with only 14.5% in group C patients, finding a reflux more than 5ml/s (determined by VFI) in 26.1% of the group C patients, comparing with only 4% of group B patients (p<0.05). We have shown that in patients with GSV reflux those with incompetence of the ostial valve of the GSV show greater venous reflux and dilatation of the saphenous trunk than those in whom the ostial valve is competent.


Immune gene co-expression signatures implicated in occurence and persistence of cognitive dysfunction in depression.
Cognitive dysfunction contributes significantly to the burden caused by Major Depressive Disorder (MDD). Yet, while compelling evidence suggests that different biological processes play a part in both MDD aetiology and the development of cognitive decline more generally, we only begin to understand the molecular underpinnings of depression-related cognitive impairment. Developments in psychometric assessments, molecular high-throughput methods and systems biology derived analysis strategies advance this endeavour. Here, we aim to identify gene expression signatures associated with cognitive dysfunction and cognitive improvement following therapy using RNA sequencing to analyze the whole blood-derived transcriptome of altogether 101 MDD patients who enrolled in the CERT-D study. The mRNA(Nova)Seq based transcriptome was analyzed from whole blood taken at baseline assessment, and patients' cognitive performance was measured twice at baseline and following eight weeks of therapy by means of the THINC integrated tool. Thirty-six patients showed comparatively low cognitive performance at baseline assessment, and 32 patients showed comparatively strong cognitive improvement following therapy. Differential gene expression analysis was performed using limma to a significance threshold of 0.05 and a logFC cutoff of |1.2|. Although we observed some indications for expression differences related to low cognitive performance and cognitive therapy response, signals did not withstand adjustment for multiple testing. Applying WGCNA, we retrieved altogether 25 modules of co-expressed genes and we used a combination of correlational and linear analyses to identify modules related to baseline cognitive performance and cognitive improvement following therapy. Three immune modules reflected distinct but interrelated immune processes (the yellow module: neutrophil-mediated immunity, the darkorange module: interferon signaling, the tan module: platelet activation), and higher expression of the yellow (r = -0.21, p < .05), the dark orange (r = 0.2, p < .05), and the tan (r = -0.23, p < .05) module correlated significantly negatively with patients' cognitive baseline performance. Patients' cognitive baseline performance was a significant predictor of the darkorange module (b = -0.039, p < .05) and the tan module's expression (b = 0.02, p < .05) and was close to becoming a significant predictor of the yellow module's expression (b = -0.02, p = .05). Furthermore, patients characterized by comparatively low cognitive performance at baseline showed significantly higher expression of the tan module when compared to all other patients F(1,97) = 4.32, p < .05, η= 0.04. Following eight weeks of treatment, we observed altogether significant improvement in patients' cognitive performance (b = 0.30, p < .001), and patients with comparatively high cognitive gain showed noticeably lower, but not significantly lower F(1,98) = 3.76, p = .058, expression of a dark turquoise module, which reflects complement and B-cell-associated immune processes. Noteworthy, the relation between cognitive performance and module expression remained observable after controlling for symptom severity and BMI, which partly accounted for variance in module expression. As such, our findings provide further evidence for the involvement of immune processes in MDD related cognitive dysfunction and they suggest that different immune processes contribute to the development and long-term persistence of cognitive dysfunction in the context of depression.


Enforcement of developmental lineage specificity by transcription factor Oct1.
Embryonic stem cells co-express Oct4 and Oct1, a related protein with similar DNA-binding specificity. To study the role of Oct1 in ESC pluripotency and transcriptional control, we constructed germline and inducible-conditional Oct1-deficient ESC lines. ESCs lacking Oct1 show normal appearance, self-renewal and growth but manifest defects upon differentiation. They fail to form beating cardiomyocytes, generate neurons poorly, form small, poorly differentiated teratomas, and cannot generate chimeric mice. Upon RA-mediated differentiation, Oct1-deficient cells induce lineage-appropriate developmentally poised genes poorly while lineage-inappropriate genes, including extra-embryonic genes, are aberrantly expressed. In ESCs, Oct1 co-occupies a specific set of targets with Oct4, but does not occupy differentially expressed developmental targets. Instead, Oct1 occupies these targets as cells differentiate and Oct4 declines. These results identify a dynamic interplay between Oct1 and Oct4, in particular during the critical window immediately after loss of pluripotency when cells make the earliest developmental fate decisions.


Norepinephrine increases the pathogenic potential of Campylobacter jejuni.
Campylobacter jejuni can cause a spectrum of diseases in humans, ranging from enteritis and diarrhoea to severe inflammation, profuse bloody diarrhoea and chronic relapsing infection. Norepinephrine (NE) levels in the intestine increase under conditions of stress and trauma, and are thought to result in spill over of NE into the intestinal lumen. NE is known to stimulate the growth of a range of bacterial species, and to increase the pathogenicity of Escherichia coli. To determine the effects of NE on the pathogenic potential of C jejuni in a model system. C jejuni was grown in iron-replete and iron-limited media in the presence and absence of 100 microM NE. Several virulence-associated characteristics, including motility and cell invasion, were measured. When C jejuni was grown in iron-limited media in the presence of NE, growth rate, motility and invasion of cultured epithelial cells were increased compared with cultures grown in the absence of NE. Bacteria exposed to NE during growth also caused greater subsequent disruption of cultured epithelial cell monolayers, inducing widespread breakdown of tight junctions. Exposure to NE causes an increase in the virulence-associated properties of Campylobacter. Stress and concomitant infection could therefore be contributory factors to the variable presentation of this disease.


Continuous clinical improvement of mild-to-moderate seborrheic dermatitis and rebalancing of the scalp microbiome using a selenium disulfide-based shampoo after an initial treatment with ketoconazole.
Scalp seborrheic dermatitis (SD) is a chronic, relapsing, and inflammatory scalp disease. Studies indicate a global bacterial and fungal microbiota shift of scalp SD, as compared to healthy scalp. Ketoconazole and selenium disulfide (SeS2 ) improve clinical signs and symptoms in both scalp dandruff and SD. The main objective of this study was to investigate the changes in the scalp microbiota diversity and counts in subjects with scalp SD during a two-phase treatment period. The scalp microbiota and clinical efficacy were investigated in 68 subjects with mild-to-moderate scalp SD after an initial one-month treatment with 2% ketoconazole, and after a 2-month maintenance phase, either with a 1% SeS2 -based shampoo or its vehicle. Thirty one subjects in the active and 37 subjects in the vehicle group participated. Ketoconazole provided an improvement of clinical symptoms (adherent (-1.75 p < 0.05), non-adherent (-1.5, p < 0.05)) flakes and erythema (scores 1.67-0.93, p < 0.001), in an increased fungal diversity and in a significant (p < 0.005) decrease of Malassezia spp. SeS2 provided an additional clinical improvement (-0.8; p = 0.0002 and -0.7; p = 0.0081 for adherent and non-adherent flakes, respectively, at Day 84) compared to the vehicle associated with a low Malassezia spp. count and an additional significant (p < 0.001) decrease of the Staphylococcus spp. level. Selenium disulfide provides an additional benefit on the scalp microbiota and in clinical symptoms of SD and dandruff after treatment with ketoconazole. The results confirm the role of Staphylococcus spp. in scalp SD and open possible perspectives for preventing relapses.


[Workplace social capital and intention to stay among Chinese nurses: a structural equation model].
Objective: To explore a model that workplace social capital is associated with intention to stay (ITS) in the nursing profession and that this association is partially mediated by organizational commitment, job satisfaction, and job stress among Chinese nurses. Methods: A cross-sectional, observationalstudy was conducted in Shanghai, China between September and December 2014. Two thousandforty-two nurses from 23 healthcare organizations were recruited for the current study using a two-stage sampling process.Intention to stay, workplace social capital, job satisfaction, organizational commitment, and job stress was measured by validated scale. Measured variable path analysis (MVPA) was used to test their hypothesized relationships. Results: There were significant positive direct effects from workplace social capital (β=0.11, P<0.01) , organizational commitment (β=0.81, P<0.01) and job satisfaction (β=0.03, P<0.01) to ITS, and a negative direct effects from job strain to ITS (β=-0.03, P<0.01) . The model explained 84% of the variability in ITS. Additionally, workplace social capital had significant positive direct effects on organizational commitment (β=0.65, P<0.01) , job satisfaction (β=0.44, P<0.01) and negative direct effects on job strain (β=-0.35, P<0.01) . The indirect effect of social capital to ITS was 0.55. Job satisfaction was positively associated with organizational commitment (r=0.47, P<0.01) , and negtively associated with job stress (r=-0.12, P<0.01) . Job stress was negtively associated with organizational commitment (r=-0.20, P<0.01) . Conclusion: This study suggests that greater workplace social capital may lead to higher ITS in nursing primarily by increasing commitment to the nursing occupation and their job satisfaction and by reducing their sense of job stress.


Impact of meningococcal vaccination on carriage and disease transmission: A review of the literature.
Colonization of the human nasopharyngeal tract by the bacterium Neisseria meningitidis is usually asymptomatic, but life-threatening meningococcal disease with a clinical presentation of meningitis, septicemia, or more rarely, gastrointestinal symptoms, can develop. Invasive meningococcal disease (IMD) can be fatal within 24 hours, but IMD is vaccine-preventable. Vaccines used to protect against IMD caused by 5 of the 6 most common serogroups (A, B, C, W, and Y) may also influence carriage prevalence in vaccinated individuals. Lower carriage among vaccinated people may reduce transmission to nonvaccinated individuals to provide herd protection against IMD. This article reviews observational and clinical studies examining effects of vaccination on N. meningitidis carriage prevalence in the context of mass vaccination campaigns and routine immunization programs. Challenges associated with carriage studies are presented alongside considerations for design of future studies to assess the impact of vaccination on carriage.


Indium-111 antimyosin scintigraphy to assess myocardial damage in patients with suspected myocarditis and cardiac rejection.
Indium-111 antimyosin scans were used to assess myocardial damage in patients with suspected myocarditis and cardiac transplant rejection. The calculation of a myocardium to lung ratio (AM index) to quantify antimyosin uptake was performed. AM index in normal subjects (n = 8) at 48 hr postinjection was 1.46 +/- 0.04. In patients with suspected myocarditis (16 studies in 13 patients), AM index was 2.0 +/- 0.5 (p less than 0.001); suggesting a considerable incidence of ongoing cell damage in this group, despite the small proportion of positive right ventricular endomyocardial biopsy (RVbx) (4/13). In patients studied after cardiac transplantation (37 studies in 17 patients), AM indexes correlated with RVbx. In patients with RVbx proven rejection (n = 14), AM index was 1.87 +/- 0.19 (p less than 0.001). In patients with RVbx showing infiltrates but not myocyte damage (n = 13), AM index was 1.80 +/- 0.27 (p = 0.02). In patients with normal RVbx (n = 10), AM index was 1.56 +/- 0.17 (p = NS versus controls; p = 0.001 versus those with positive RVbx). Calculated AM indexes correlated with graded visual analysis of the scans (r = 0.823; p = 0.001). Antimyosin scans are an appropriate method to assess myocardial damage in patients with suspected myocarditis and cardiac rejection.


Prion removal by nanofiltration under different experimental conditions.
Manufacturing processes used in the production of biopharmaceutical or biological products should be evaluated for their ability to remove potential contaminants, including TSE agents. In the present study, we have evaluated scrapie prion protein (PrP Sc) removal in the presence of different starting materials, using virus removal filters of different pore sizes. Following 75 nm filtration, PrP Sc was detected in the filtrate by Western blot (WB) analysis when a "super-sonicated" microsomal fraction derived from hamster adapted scrapie strain 263K (263K MF) was used as the spike material. In contrast, no PrP Sc was detected when an untreated 263K MF was used. By using spike materials prepared in a manner designed to optimize the particle size distribution within the preparation, only 15 nm filtration was shown to remove PrP Sc to below the limits of detection of the WB assays used under all the experimental conditions. However, infectious PrP Sc was recovered following 15 nm filtration under one experimental condition. The results obtained suggest that the nature of the spike preparation is an important factor in evaluating the ability of filters to remove prions, and that procedures designed to minimize the particle size distribution of the prion spike, such as the "super-sonication" or detergent treatments described herein, should be used for the preparation of the spike materials.


Coordinated and Iterative Enzyme Catalysis in Fungal Polyketide Biosynthesis.
Fungal polyketides are natural products with great chemical diversity that exhibit a wide range of biological activity. This chemical diversity stems from specialized enzymes encoded in the biosynthetic gene cluster responsible for the natural product biosynthesis. Fungal polyketide synthases (PKS) are the megasynthases that produce the carbon scaffolds for the molecules. Subsequent downstream tailoring enzymes such as oxygenases will then further modify the organic framework. In fungi, many of these enzymes have been found to work iteratively-catalyzing multiple reactions on different sites of the substrate. This perspective will analyze several examples of fungal polyketides that are assembled from a scaffold-building iterative PKS and an accompanying iterative tailoring oxygenase. In these examples, the PKS product is designed for downstream iterative oxygenations to generate additional complexity. Together, these iterative enzymes orchestrate the efficient biosynthesis of elaborate natural products such as lovastatin, chaetoglobosin A, cytochalasin E, and aurovertin E.


XLID-causing mutations and associated genes challenged in light of data from large-scale human exome sequencing.
Because of the unbalanced sex ratio (1.3-1.4 to 1) observed in intellectual disability (ID) and the identification of large ID-affected families showing X-linked segregation, much attention has been focused on the genetics of X-linked ID (XLID). Mutations causing monogenic XLID have now been reported in over 100 genes, most of which are commonly included in XLID diagnostic gene panels. Nonetheless, the boundary between true mutations and rare non-disease-causing variants often remains elusive. The sequencing of a large number of control X chromosomes, required for avoiding false-positive results, was not systematically possible in the past. Such information is now available thanks to large-scale sequencing projects such as the National Heart, Lung, and Blood (NHLBI) Exome Sequencing Project, which provides variation information on 10,563 X chromosomes from the general population. We used this NHLBI cohort to systematically reassess the implication of 106 genes proposed to be involved in monogenic forms of XLID. We particularly question the implication in XLID of ten of them (AGTR2, MAGT1, ZNF674, SRPX2, ATP6AP2, ARHGEF6, NXF5, ZCCHC12, ZNF41, and ZNF81), in which truncating variants or previously published mutations are observed at a relatively high frequency within this cohort. We also highlight 15 other genes (CCDC22, CLIC2, CNKSR2, FRMPD4, HCFC1, IGBP1, KIAA2022, KLF8, MAOA, NAA10, NLGN3, RPL10, SHROOM4, ZDHHC15, and ZNF261) for which replication studies are warranted. We propose that similar reassessment of reported mutations (and genes) with the use of data from large-scale human exome sequencing would be relevant for a wide range of other genetic diseases.


Impact of timing and duration of postoperative delirium: a retrospective observational study.
Recent reports suggest that not all critically ill patients with delirium share the same consequences. The outcomes of surgical intensive care unit patients with postoperative delirium were evaluated depending on the onset and duration of delirium. A total of 527 patients who were admitted from the operating theater and cared for in the surgical intensive care unit for >24 hours were evaluated for delirium using the Confusion Assessment Method for intensive care unit, 3 times a day. Patients were analyzed according to the onset time and duration of delirium. Patients were classified into 4 groups according to the onset and duration of delirium: no delirium, early brief delirium (delirium for <1 day on postoperative day 0), late brief delirium (delirium for <1 day after postoperative day 0), and persistent delirium (delirium for ≥1 days). Duration of stay (intensive care unit and hospital) and mortality (intensive care unit, hospital, and 1-year) were outcomes of interest. Of the 527 patients, delirium developed in 119 (22.6%) patients. More than two-thirds of the patients developed delirium on postoperative day 0 or 1, and 70% of patients developed delirium for >24 hours (persistent). Persistent delirium was associated with longer intensive care unit (4.6 [1.1-53.3] vs 1.6 [1.1-37.5] days) and hospital duration of stay (24 [3-112] vs 16 [2-225] days) and higher hospital mortality (14.5% vs 2.2%) compared to no delirium (P < .01). For postoperative intensive care unit patients, intensive care unit and hospital duration of stay did not seem to differ between patients with early brief delirium or no delirium, whereas patients with late brief or persistent delirium seemed to show longer intensive care unit and hospital duration of stay and higher mortality.


Raman spectroscopy as a process analytical technology tool for the understanding and the quantitative in-line monitoring of the homogenization process of a pharmaceutical suspension.
The aim of this study was to propose a Process Analytical Technology (PAT) strategy for the quantitative in-line monitoring of an aqueous pharmaceutical suspension using Raman spectroscopy. A screening design was used to study the significance of process variables (mixing speed and height of the stirrer in the reactor) and of formulation variables (concentration of the active pharmaceutical ingredient (API) ibuprofen and the viscosity enhancer (xanthan gum)) on the time required to homogenize an aqueous pharmaceutical model suspension as response variable. Ibuprofen concentration (10% and 15% (w/v)) and the height of stirrer (position 1 and 2) were discrete variables, whereas the viscosity enhancer (concentration range: 1-2 g L-1) and the mixing speed (700-1000 rpm) were continuous variables. Next, a multilevel full factorial design was applied to study the effect of the remaining significant variables upon the homogenization process and to establish the optimum conditions for the process. Interactions between these variables were investigated as well. During each design experiment, the conformity index (CI) method was used to monitor homogeneity of the suspension mixing system in real-time using Raman spectroscopy in combination with a fibre optical immersion probe. Finally, a principal component regression (PCR) model was developed and evaluated to perform quantitative real-time and in-line measurements of the API during the mixing process. The experimental design results showed that the suspension homogenization process is an irregular process, for which it is impossible to model the studied variables upon the measured response variable. However, applying the PCR model it is possible to predict in-line and real-time the concentration of the API in a suspension during a mixing process. In this study, it is shown that Raman spectroscopy is a suitable PAT tool for the control of the homogenization process of an aqueous suspension. Raman spectroscopy not only allowed real-time monitoring of the homogeneity of the suspension, but also helped (in combination with experimental design) to understand the process. Further, the technique allowed real-time and in-line quantification of the API during the mixing process.


Transformation between Inverse Bicontinuous Cubic Phases of a Lipid from Diamond to Gyroid.
The transformation between inverse bicontinuous cubic phases of a lipid from diamond (QII(D)) to gyroid (QII(G)) in the single crystal region of monoolein was studied. X-ray diffraction data indicate that the single orientation of the QII(D) phase was converted into an almost single orientation of the QII(G) phase. The [111] and [11̅0] directions of a single crystal of the QII(D) phase corresponded to the [202] and [04̅0] directions of the QII(G) phase, respectively. This orientation relationship indicated that one direction in the four-branched water channels of the QII(D) phase was preserved in the three-branched water channels of the QII(G) phase. Using this relationship, a transformation model was constructed in which one direction of the water channels was preserved while another direction appeared.


Long-Term Assessment of AAV-Mediated Zinc Finger Nuclease Expression in the Mouse Brain.
Gene editing tools like TALENs, ZFNs and Crispr/Cas now offer unprecedented opportunities for targeted genetic manipulations in virtually all species. Most of the recent research in this area has concentrated on manipulation of the genome in isolated cells, which then give rise to transgenic animals or modified stem cell lines. Much less is known about applicability of genetic scissors in terminally differentiated, non-dividing cells like neurons of the adult brain. We addressed this question by expression of a pair of ZFNs targeting the murine cathepsin D gene in CNS neurons by means of an optimized AAV viral vector. We show that ZFN expression resulted in substantial depletion of cathepsin D from neuronal lysosomes, demonstrating a robust gene deletion. Importantly, long-term ZFN expression in CNS neurons did not impair essential neuronal functionality and did not cause inflammation or neurodegeneration, suggesting that potent genetic scissors can be expressed safely in the mouse brain. This finding opens up new venues to create novel research models for neurodegenerative disorders.


Twice-daily insulin degludec/insulin aspart provides superior fasting plasma glucose control and a reduced rate of hypoglycaemia compared with biphasic insulin aspart 30 in insulin-naïve adults with Type 2 diabetes.
To evaluate the efficacy and safety of twice-daily insulin degludec/insulin aspart vs. twice-daily biphasic insulin aspart 30 in people with Type 2 diabetes mellitus who were naïve to insulin. In this 26-week, multinational, open-label, controlled, two-arm, parallel-group, treat-to-target trial, participants [mean (± sd) age 58.9 (±8.9) years, duration of diabetes 9.5 (±5.9) years, HbA1c 68 (±8.7) mmol/mol or 8.4 (±0.8)% and BMI 31.2 (±4.2) kg/m(2) ) were randomized (1:1) to insulin degludec/insulin aspart (n = 197) or biphasic insulin aspart 30 (n = 197), administered with breakfast and the main evening meal, titrated to a self-monitored plasma glucose target > 3.9 and ≤ 5.0 mmol/l. The mean HbA1c was reduced to 49 mmol/mol (6.6%) with insulin degludec/insulin aspart and 48 mmol/mol (6.5%) with biphasic insulin aspart 30. Insulin degludec/insulin aspart achieved the prespecified non-inferiority margin (estimated treatment difference 0.02%; 95% CI -0.12, 0.17). Insulin degludec/insulin aspart was superior in lowering fasting plasma glucose (estimated treatment difference -1.00 mmol/l; 95% CI -1.4, -0.6; P < 0.001) and reducing overall and nocturnal confirmed hypoglycaemia at a similar overall insulin dose compared with biphasic insulin aspart 30. Similar proportions of participants in each arm experienced severe hypoglycaemia. Adverse events were equally distributed. Consistent with previous findings, insulin degludec/insulin aspart twice daily effectively improved long-term glycaemic control, with superior reductions in FPG, and significantly less overall and nocturnal confirmed hypoglycaemia compared with biphasic insulin aspart 30 in people with Type 2 diabetes who were insulin-naïve.


Omicron variant as nature's solution to the COVID-19 pandemic.
Omicron is a variant of the COVID-19 virus that is causing considerable concern worldwide, with an increasing number of countries re-imposing national lockdowns. Our objective is to comment on its impact and to suggest that, threatening as it is, Omicron may well contribute to a resolution of the current pandemic. On 31 December 2019, the World Health Organization (WHO) reported on a cluster of cases of pneumonia in Wuhan, China. Soon after, Chinese investigators who made the discovery identified the causative virus as a new coronavirus, now known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). An effective vaccine was licenced for emergency use within a year of its first sequencing. SARS-CoV-2, in common with many respiratory viruses, mutates rapidly, and the challenge for vaccine developers is to obtain vaccines that are effective against the new variants. The licenced first-generation vaccines were fortunately all highly effective against the variant known as Delta. The variant of greatest current concern is the Omicron variant, a highly infectious agent, which seems to show a significant vaccine escape with existing vaccines. Infection protects against further infection. If Omicron turns out to cause less severe disease, it may well be a contributor to ending the pandemic. It is unlikely that the available vaccines will bring rapid control of the current pandemic, given their patchy availability worldwide and the residual pool of unvaccinated people. New vaccines take time to develop and to deploy even in the age of mRNA vaccines. If Omicron turns out to be relatively mild, it may well be that when we look back at the history of the current pandemic, the variant would be seen as a contributor to its solution. The hand of nature may well show more largesse than the developed nations in immunizing the world.


Age effects on body fluid distribution during exercise in the heat.
The purpose was to study the effects of age on body fluid distribution during exercise and exercise combined with heat stress. Ten young (Y; 23.2 +/-0.7 yr) and eight older (O; 65.3 +/- 1.0 yr) men performed two 80-min intermittent exercise bouts on a cycle ergometer at 50% VO2peak, one in a thermoneutral (N), 22 degrees C environment, the other in a hot (H), 40 degrees C environment. For each condition, changes in serum protein, albumin, and electrolytes (Cl-, Na+, K+); total body water (TBW); plasma volume (PV); and interstitial (ISF), extracellular (ECF), and intracellular (ICF) fluids were determined. During exercise, [Cl-] responses increased more in O than Y (p < or = 0.05) while albumin (g) increased in Y and decreased in O (p < or = 0.05). There were no age-related differences in total protein changes nor any of the other blood parameters. PV decreased similarly for all subjects in both conditions with larger decreases in the H environment (241.8 +/- 40.3 vs. 478.3 +/- 46.0 mL). Loss of TBW was exacerbated in the H condition and significantly different (p < 0.05) between the groups (N: Y = -1.03 +/- 0.076 L, O = -0.88 +/- 0.12 L; H: Y = -1.65 +/- 0.12 L, O = -1.85 +/- 0.14 L). The O group lost more ICF (p < 0.05) (N: Y = -510.4 +/- 86.8 mL, O = -631.7 +/- 115.1 mL; H: Y = -529.3 +/- 118.0 mL, O = -928.6 +/- 118.9 mL) and less ISF (p < 0.05) (N: Y = -295.5 +/- 101.1 mL, O = 15.8 +/- 77.6 mL; H: Y = -684.6 +/- 134.5 mL, O = -397.9 +/- 165.5 mL) in both conditions. There was an interactive effect between age and heat stress in TBW loss. In addition, older individuals lost more ICF and less ISF than younger individuals during prolonged exercise. These findings suggest that the utilization of ICF, rather than ISF, to preserve PV during cycling exercise at or near 50% VO2max may be an age-related compensation.


[Portal hypertensive biliopathy--an unusual cause of obstructive jaundice].
Biliary symptoms may be an unusual presentation of portal hypertension, especiaLly in patients with portal vein thrombosis. This is a case report of a patient who developed acute and chronic portal vein thrombosis that presented as obstructive jaundice. The discussion will focus on diagnostic modalities and therapeutic options.


Molecular characterization of Listeria monocytogenes of the serotype 4b complex (4b, 4d, 4e) from two turkey processing plants.
Most foodborne outbreaks of listeriosis have been found to involve a small number of closely related strains of Listeria monocytogenes serotype 4b. The ecology of these organisms and their reservoirs in nature or in the processing plant environment, however, remain poorly understood. Surveys of environmental samples from two turkey processing plants in the United States indicated presence of L. monocytogenes of the serotype 4b complex (serotype 4b and the closely related serotypes 4d and 4e). In addition, environmental and raw product samples from one plant repeatedly yielded isolates with genetic markers typical of two major serotype 4b epidemic clonal groups, ECI and ECII. The pulsed field gel electrophoresis (PFGE) profiles of these isolates, however, were clearly distinct from those of confirmed epidemic-associated strains. Furthermore, we observed minor but consistent differences in PFGE profiles of isolates that harbored ECI- or ECII-specific genetic markers, and that were obtained at different sampling times from the same plant. The findings suggest processing plant persistence (or repeated introductions) and genomic diversification of L. monocytogenes serotype 4b isolates that harbor ECI- or ECII-specific genetic markers. Such diversification would need to be taken into consideration in further efforts to elucidate the evolution and epidemiology of these organisms.


MiR-634 sensitizes nasopharyngeal carcinoma cells to paclitaxel and inhibits cell growth both in vitro and in vivo.
Resistance to chemotherapy is one of the key causal factors in cancer death and increasing evidence has revealed that microRNAs (miRNAs) are involved in chemoresistance in many kinds of human cancers. Paclitaxel has been used for treatment of advanced nasopharyngeal carcinoma (NPC); however, treatment failure often occurs due to development of acquired paclitaxel resistance. In this study, based on miRNA microarray screening and qRT-PCR validation, we found six differentially expressed miRNAs in our induced paclitaxel-resistant NPC CNE-1/Taxol cells. Furthermore, we clarified the role of miR-634, most significantly downregulated in the paclitaxel-resistant CNE-1/Taxol, in regulating the paclitaxel sensitivity in NPC cells. We restored miR-634 expression in the CNE-1/Taxol cells by lentivirus infection, and found restoration of miR-634 re-sensitized the CNE-1/Taxol cells to paclitaxel in vitro by MTT assay and colony formation assay. In xenograft mouse model, we found that miR-634 inhibited tumor growth and enhanced paclitaxel sensitivity. Thus, our findings provide important information for the development of targeted gene therapy for reversing paclitaxel resistance in NPC.


Pattern of malaria in hospitalized children in Khartoum state.
Malaria remains a major health problem in Sudan with significant morbidity and mortality particularly in children. We prospectively studied children with malaria admitted to an Emergency Department in Khartoum (August-November 2014). Malaria diagnosis was based on a positive blood smear and rapid diagnostic test. The aim was to study the clinical and laboratory features and short-term outcome of malaria among hospitalized children. Data collected from 112 children (males; 56.3%) who fulfilled the criteria for diagnosis of malaria of whom 72.3% had severe malaria and 27.7% uncomplicated malaria (UM). The mean age was 69.2 ± 54.5 months. Hyperparasitemia was detected in 53% of positive blood smears. Plasmodium falciparum was detected in 69.4%, P. vivax in 26.5%, and mixed species in 4.1%. The risk of severe malaria was significantly higher in patients with hyperparasitemia and P. vivax infection (P = 0.001 and P = 0.014 respectively). Severe malaria cases had significantly higher prevalence of thrombocytopenia and lower mean platelet count than those with UM, P = 0.001 each. Serious complications of severe malaria were cerebral malaria, severe malaria anaemia and acute kidney injury (AKI). The overall case fatality rate was 5.3% and that from severe disease was 4.9%. All deaths were among <60 months-olds and were due to P. falciparum infection with AKI being the only significant risk factor for death (P = 0.045). In Khartoum state, UM is still an important cause of morbidity in children. P. vivax has emerged as a causative species of severe malaria. The lower mortality rate of malaria probably reflects improvement in health care.


G protein- and β-arrestin Signaling Profiles of Endothelin Derivatives at the Type A Endothelin Receptor.
Endothelin-1 (ET-1) is a potent vasoconstrictor in the cardiovascular system, an effect mediated through the type A endothelin receptor (ETAR), a G protein-coupled receptor (GPCR). Antagonists of the ETAR have shown promising results in randomized clinical trials. However, side effects limit widespread use. Biased agonists have been developed to mitigate the untoward effects of a number of GPCR antagonists. These agents block deleterious G-coupled pathways while stimulating protective β-arrestin pathways. The goal of this study was to test whether there was any significant ligand bias between endothelin derivatives, and whether this could have any physiologic effects in the cardiovascular system. A panel of endothelin derivatives were tested in assays of G protein signaling and β-arrestin 2 recruitment at the ETAR. We then tested the effects of ET-1 on the vasopressor response in wild-type and β-arrestin 1 and 2 KO mice. We found the endothelins activated a wide range of G proteins at the ETAR, but none of the endothelin derivatives demonstrated significant biased agonism. Endothelin derivatives did display structure-activity relationships with regards to their degrees of agonism. β-arrestin 1 and 2 knockout mice did not display any differences to wild-type mice in the acute pressor response to ET-1, and β-arrestin 2 knockout mice did not display any blood pressure differences to wild-type mice in the chronic responses to ET-1. Our findings are consistent with vasoconstriction being mediated by G proteins with a lack of significant desensitization by β-arrestins at the ETAR. These findings suggest that G protein- and β-arrestin-biased ETAR agonists could have distinct physiologic effects from balanced agonists, although the endothelin peptide scaffold does not appear suitable for designing such ligands.


Aryl Dance Reaction of Arylbenzoheteroles.
We report a 1,2-migration (aryl dance reaction) of the aryl group on heteroles. AlCl3 can efficiently convert C3-arylheteroles to C2-arylheteroles. Depending on the electron density of the substrate, conversion from C2- to C3-arylheteroles was also possible with catalytic Zn(OTf)2. A one-pot aryl dance/acylation or bromination and arylation/aryl dance cascade was also demonstrated.


Structure-based mutational analysis of ICAT residues mediating negative regulation of β-catenin co-transcriptional activity.
ICAT (Inhibitor of β-CAtenin and TCF) is a small acidic protein that negatively regulates β-catenin co-transcriptional activity by competing with TCF/LEF factors in their binding to β-catenin superhelical core. In melanoma cells, ICAT competes with LEF1 to negatively regulate the M-MITF and NEDD9 target genes. The structure of ICAT consists of two domains: the 3-helix bundle N-terminal domain binds to β-catenin Armadillo (Arm) repeats 10-12 and the C-terminal tail binds to Arm repeats 5-9. To elucidate the structural mechanisms governing ICAT/β-catenin interactions in melanoma cells, three ICAT residues Y15, K19 and V22 in the N-terminal domain, contacting hydrophobic β-catenin residue F660, were mutated and interaction was assessed by immunoprecipitation. Despite the moderate hydrophobicity of the contact, its removal completely abolished the interaction. In the ICAT C-terminal tail consensus sequence, neutralization of the electrostatic interactions between residues D66, E75 and β-catenin residues K435, K312, coupled to deletion of the hydrophobic contact between F71 and β-catenin R386, markedly reduced, but failed to abolish the ICAT-mediated negative regulation of M-MITF and NEDD9 promoters. We conclude that in melanoma cells, anchoring of ICAT N-terminal domain to β-catenin through the hook made by residue F660, trapped in the pincers formed by ICAT residues Y15 and V22, is crucial for stabilizing the ICAT/β-catenin complex. This is a prerequisite for binding of the consensus peptide to Arm repeats 5-9 and competition with LEF1. Differences between ICAT and LEF1 in their affinity for β-catenin may rely on the absence in ICAT of hydrophilic residues between D66 and F71.


Gut Microbiota Co-microevolution with Selection for Host Humoral Immunity.
To explore coevolution between the gut microbiota and the humoral immune system of the host, we used chickens as the model organism. The host populations were two lines (HAS and LAS) developed from a common founder that had undergone 40 generations of divergent selection for antibody titers to sheep red blood cells (SRBC) and two relaxed sublines (HAR and LAR). Analysis revealed that microevolution of host humoral immunity contributed to the composition of gut microbiota at the taxa level. Relaxing selection enriched some microorganisms whose functions were opposite to host immunity. Particularly, Ruminococcaceae and Oscillospira enriched in high antibody relaxed (HAR) and contributed to reduction in antibody response, while Lactobacillus increased in low antibody relaxed (LAR) and elevated the antibody response. Microbial functional analysis showed that alterations were involved in pathways relating to the immune system and infectious diseases. Our findings demonstrated co-microevolution relationships of host-microbiota and that gut microorganisms influenced host immunity.


ENDOGENOUS AND PHOTOPERIODIC DIURNAL RHYTHMS OF IN VIVO LIGHT ABSORPTION AND SCATTERING IN THE GREEN ALGA ULVA LACTUCA L.
Techniques of in vivo spectroscopy were employed to demonstrate the presence of rhythms of light absorption and scattering in the green thalloid alga Ulva lactuca L. maintained in artificial nutrient medium under constant photoperiod. The absorbance during photophase at 682 nm, the chlorophyll a in vivo absorption maximum, was typically two to three times that during scotophase. Prephased endogenous rhythms (in continued darkness) were observed for a time period equal to three photoperiods. The absorbance rhythm did not correlate with changes in extractable chlorophyll a or b concentration. No changes in linear dichroism were observed, indicating the rhythm of light absorption could not be explained on the basis of orientation of individual absorber molecules. In vivo light microscopy did not reveal differences in chloroplast orientation. Alternative mechanisms are discussed.


Protective Effects of Descurainia sophia against Gentamicin Induced Nephrotoxicity in Rats.
Several studies have tried to find an efficient agent to prevent or reverse gentamicin (Gm) induced acute kidney injury (AKI). In this study, we assessed the potential renal protective effects of Descurainia sophia (L.) Webb ex Prantl against Gm-induced nephrotoxicity in rats. Thirty-five male Wistar rats were categorized in five groups (n = 7 per group). Control group was treated with normal saline. In four experimental groups, the rats were initially treated with normal saline (A), 800 (B), 1600 (C) and 2400 (D) mg/kg Descurainia sophia respectively for 28 days. After that, the rats of experimental groups were treated with Gm (80 mg/Kg) for 7 consecutive days. Blood and urine markers, as well as apoptosis and histological features were determined. Serum BUN, creatinine, cholesterol, and triglycerides level, as well as urinary excretion of Na+ significantly increased in group A. Furthermore, Gm induced inflammatory cells infiltration, apoptosis, and renal cells injuries in rats were pretreated with normal saline (group A). However, in the rats pretreated with Descurainia sophia extract (groups B, C, and D, there were significant and dose-dependent reductions in serum BUN, creatinine, cholesterol and triglyceride, urinary Na+ excretion, apoptosis rate, and inflammatory cells infiltration in renal tissues. Overall, Descurainia sophia showed significant protective effects against Gm-induced AKI by alleviating biochemical and histological markers of renal toxicity.


Changes in serum hexachlorocyclohexane (HCH) residues in malaria spraymen after short-term occupational exposure.
The use of organochlorine insecticides for public health programs is indisputably necessary in developing countries. Thus, there is a need to evaluate human response to these chemicals under local field conditions. The effects of a short-term, 16-wk exposure to hexachlorocyclohexane has been evaluated and compared in malaria spraymen with and without any previous occupational exposure to this insecticide by measuring the pre- and post-exposure serum hexachlorocyclohexane concentration. A significant increase in original serum hexachlorocyclohexane concentration. A significant increase in original serum hexachlorocyclohexane concentration was observed in subjects on their first occupational exposure (5X), compared to those with previous exposure (3X). All workers reached approximately the same serum levels within 16 wk, regardless of their history of previous exposure.


Closing a Calix[6]arene-Based Funnel Zn2+ Complex at Its Large Rim Entrance: Consequences on Metal Ion Affinity and Host-Guest Properties.
A molecular capsule based on a calix[6]arene core closed at the small rim by a three-point coordinated metal ion and at the large rim by a three-point covalent capping is described. It is derived from a trisimidazole funnel complex capped by a trenamide unit that prevents in/out exchange of guest molecules through the large rim. A detailed comparative study with three different calixarenes provides a unique opportunity for (i) comparing the binding ability of two different coordination sites in well three-dimensional (3D)-structured macrocyclic receptors and (ii) evaluating the impact of a covalent closing of one rim of a funnel receptor while the other rim is closed by weaker coordination bonds. Indeed, this study allowed for highlighting various interesting new features. It is first shown that the trenamide site can bind a metal ion such as Zn2+ by itself. This involves a 1:1 coordination of the metal ion to the three carbonyl groups of the amide functions, which undergo trans-to-cis isomerization and are partially embedded in the calix core. When the trisimidazole core is present, the Zn2+ ion preferentially binds at the small rim, thus closing the cavity. Guest ligand exchange must then occur through a decoordination/recoordination process of the metal ion. The modification and rigidification of the calixarene conformation induced by the large rim capping strengthen the metal ion coordination at the small rim. This also leads to a selective metallo-receptor that readily binds EtNH2 under conditions where PrNH2 is not recognized at all. The increased rigidity of the receptor, however, weakens the host-guest interactions, precluding important induced-fit behaviors that are at work in the parent, large rim opened, funnel complex.


Progesterone promotes embryo adhesion by upregulating c-Fos/c-Jun transcription factor-mediated poFUT1 expression†.
The proliferation and adhesion abilities of placental trophoblasts are critical for embryo implantation and successful pregnancy. Protein O-fucosyltransferase 1 (poFUT1) and the transcription factor c-Fos/c-Jun have been found to promote trophoblastic cell invade into the endometrium. Progesterone is critical to the regulation of embryonic implantation. However, the exact role of poFUT1 in embryo proliferation and adhesion to the endometrium, and the relationship between progesterone, c-Fos/c-Jun, and poFUT1 has not been studied in detail. In the current study, we found that the serum levels of poFUT1 and progesterone significantly was decreased in miscarriage patients compared with those in normal pregnancy women, and there is a positive correlation between the changes in progesterone and poFUT1. Employing a human embryo trophoblastic cell line (JAR), we showed that progesterone facilitated the activation of c-Fos/c-Jun. Using an electrophoretic mobility shift assay and chromatin immunoprecipitation, we confirmed that the specific transcription factor c-Fos/c-Jun regulated the poFUT1 promoter, which could enhance poFUT1 transcriptional activity, thus further increasing trophoblast cell proliferation and adhesion potential. Taking these findings together, progesterone upregulates poFUT1 expression via the specific transcription factor c-Fos/c-Jun, and then increase trophoblast cell proliferation and adhesion potential. poFUT1 and progesterone may be used together as potential markers of miscarriage, and they may be diagnostic and therapeutic targets for miscarriage.


A comparison of the copper T-200 and the Lippes Loop in four countries.
A comparative field trial of the Copper TCu-200 and the Lippes loop was conducted in Colombia, Iran, Korea, and Thailand from 1971 to 1973. The study, involving 7,400 women, provided two-year first segment continuation and termination rates. In three countries, TCu-200 expulsion rates were significantly below Lippes loop expulsion rates. For women with three or fewer living children, TCu-200 expulsion rates were markedly below Lippes loop rates in all four countries. The comparative advantage accruing to the TCu-200 in this characteristic did not extend to the two-year continuation rates.


[Undifferentiated carcinoma with osteoclast-like giant cells of the pancreas. A case report].
Undifferentiated carcinoma with osteoclast-like giant cells is a rare pancreatic neoplasm, with a possible cystic pattern, includes an adenocarcinoma component and giant cell mimicking osteoclastic bone tumor. We report the case of a 72-year woman admitted for jaundice and epigastric pain. Abdominal ultrasound and computed tomography scan revealed a 6 x 5 cm cystic and solid tumor of the pancreatic head with both bile duct and pancreatic duct dilatation. A pancreaticoduodenectomy was performed and pathologic examination revealed an undifferentiated carcinoma with osteoclast-like giant cells of the pancreatic head. The patient was alive and disease-free with a 18-month follow-up.


Synchronization states and multistability in a ring of periodic oscillators: experimentally variable coupling delays.
We experimentally study the complex dynamics of a unidirectionally coupled ring of four identical optoelectronic oscillators. The coupling between these systems is time-delayed in the experiment and can be varied over a wide range of delays. We observe that as the coupling delay is varied, the system may show different synchronization states, including complete isochronal synchrony, cluster synchrony, and two splay-phase states. We analyze the stability of these solutions through a master stability function approach, which we show can be effectively applied to all the different states observed in the experiment. Our analysis supports the experimentally observed multistability in the system.


A Naïve Phage Display Library-Derived Nanobody Neutralizes SARS-CoV-2 and Three Variants of Concern.
The emergence of the coronavirus disease 2019 (COVID-19) pandemic and the new severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) variants of concern (VOCs) requires the continuous development of safe, effective, and affordable prevention and therapeutics. Nanobodies have demonstrated antiviral activity against a variety of viruses, providing a new candidate for the prevention and treatment of SARS-CoV-2 and its variants. SARS-CoV-2 glycoprotein spike 1 subunit (S1) was selected as the target antigen for nanobody screening of a naïve phage display library. We obtained a nanobody, named Nb-H6, and then determined its affinity, inhibition, and stability by ELISA, Competitive ELISA, and Biolayer Interferometry (BLI). Infection assays of authentic and pseudotyped SARS-CoV-2 were performed to evaluate the neutralization of Nb-H6. The structure and mechanism of action were investigated by AlphaFold, docking, and residue mutation assays. We isolated and characterized a nanobody, Nb-H6, which exhibits a broad affinity for S1 and the receptor binding domain (RBD) of SARS-CoV-2, or Alpha (B.1.1.7), Delta (B.1.617.2), Lambda (C.37), and Omicron (BA.2 and BA.5), and blocks receptor angiotensin-converting enzyme 2 (ACE2) binding. Moreover, Nb-H6 can retain its binding capability after pH or thermal treatment and effectively neutralize both pseudotyped and authentic SARS-CoV-2, as well as VOC Alpha (B.1.1.7), Delta (B.1.617.2), and Omicron (BA.2 and BA.5) pseudoviruses. We also confirmed that Nb-H6 binds two distinct amino acid residues of the RBD, preventing SARS-CoV-2 from interacting with the host receptor. Our study highlights a novel nanobody, Nb-H6, that may be useful therapeutically in SARS-CoV-2 and VOC outbreaks and pandemics. These findings also provide a molecular foundation for further studies into how nanobodies neutralize SARS-CoV-2 and variants and imply potential therapeutic targets for the treatment of COVID-19.


Addressing tobacco in managed care: a survey of dentists' knowledge, attitudes, and behaviors.
This study assessed the tobacco cessation knowledge, attitudes, and behaviors of dentists participating in a large managed care dental plan. Participating dentists in 4 states were surveyed via mail. Dentists' perceived success in helping patients quit using tobacco was highly correlated with discussion of specific strategies for quitting, advice about the use of nicotine gum, and time spent counseling patients. Dentists who were confident about their smoking cessation knowledge frequently advised patients to quit and spent more time counseling patients about tobacco cessation. Tobacco cessation is not a routine part of dental practice. Knowledge, time spent counseling patients, and specific strategies for quitting were associated with dentists' perceptions of success.


Low energy X-ray (grenz ray) treatment of purified islets prior to allotransplant markedly decreases passenger leukocyte populations.
Grenz rays, or minimally penetrating X-rays, are known to be an effective treatment of certain recalcitrant immune-mediated skin diseases, but their use in modulating allograft rejection has not been tested. We examined the capacity of grenz ray treatment to minimize islet immunogenicity and extend allograft survival in a mouse model. In a preliminary experiment, 1 of 3 immunologically intact animals demonstrated long-term acceptance of their grenz ray treated islet allograft. Further experiments revealed that 28.6% (2 of 7) grenz ray treated islet allografts survived >60 d. A low dose of 20Gy, was important; a 4-fold increase in radiation resulted in rapid graft failure, and transplanting a higher islet mass did not alter this outcome. To determine whether increased islet allograft survival after grenz treatment would be masked by immunosuppression, we treated the recipients with CTLA-4 Ig, and found an additive effect, whereby 17.5% more animals accepted the graft long-term versus those with CTLA-4 Ig alone. Cell viability assays verified that islet integrity was maintained after treatment with 20Gy. As well, through splenocyte infiltration analysis, donor CD4+ T cell populations 24-hours after transplant were decreased by more than16-fold in recipients receiving irradiated islets compared with control. Donor CD8+ T cell populations, although less prevalent, decreased in all treatment groups compared with control. Our results suggest that brief treatment of isolated islets with low energy grenz rays before allotransplantation can significantly reduce passenger leukocytes and promote graft survival, possibly by inducing donor dendritic cells to differentiate toward a tolerogenic phenotype.


Immunoreactivity of PTH-binding in intact bovine kidney tissue and cultured cortical kidney cells indicative for specific receptors.
Localization of PTH-binding sites has been examined in intact kidney sections and cultured cells derived from bovine kidney cortex. Tissue sections were incubated with 10(-7) M bovine PTH (1-84) for 2 h, cells for 15 min, at 37 degrees C. Visualization of PTH-binding was achieved by immunocytochemistry using a carboxy-terminal specific anti-PTH antiserum (S 478). For control, cell culture incubations were performed applying competitively 10(-7) bovine PTH (1-84) and a 10-fold excess of synthetic 1-34 PTH fragment, not antigenic for S 478. This resulted in a lack of staining. PTH-binding was found in all cells of the proximal and the distal tubule, and with less intensity in the thick ascending limb of Henle's loop. In collecting ducts a PTH specific staining was also present, which was confined to single cells localized between others without PTH binding sites. No staining was seen in glomerula, the thin limb of Henle's loop, in blood vessels, and in connective tissue. The data suggest that large parts of the nephron contain PTH-binding sites, although in different amounts. This is in agreement with the numerous actions of PTH in the kidney. In the collecting segment a distinct cell-to-cell difference was disclosed indicative for different functional states or cellular heterogeneity.


A method of estimating intracranial decompensation in man.
A new method of estimating intracranial decompensation in man is described. An on-line computer system is connected to an intracranial pressure (ICP) monitoring system to compute regression plots of mean ICP vs standard deviation; standard deviation is used as a measure of ICP instability. Two zones with distinctly different slopes are a characteristic feature of these plots. It is thought that the changes of slope signify intracranial decompensation.


Semi-automated Tip Snip cloning of restriction fragments into and out of plasmid polylinkers.
Synthetic biologists rely on semi-synthetic recombinant plasmids, but DNA synthesis is constrained by practical limits on length, accuracy, and sequence composition. Cloned DNA parts can be assembled into longer constructs via subcloning, but conventional methods are labor-intensive. One-pot recombination reactions are more convenient but harder to troubleshoot, and those that depend on PCR to create fragments with compatible ends necessitate re-sequencing. The Tip Snip protocol described here enables the subcloning of an insert from one plasmid polylinker into another without PCR or gel purification steps. Cohesive ends of unwanted restriction fragments are snipped off by additional restriction endonucleases. The resulting short fragments (snippets) are eliminated by hybridization to complementary oligonucleotides (anti-snippets) and subsequent size-selection spin-column chromatography. Unwanted linear donor vectors are ligated to double-stranded oligonucleotides (unlinkers) so that only the desired insert and recipient plasmid form circular DNA capable of transforming bacteria. This new method is compatible with high-throughput processing and automated liquid handling, and because no specialized vectors, reagents, selection schemes, or analytical techniques are required, the barriers to adoption are low.


The role of allochrony in speciation.
The importance of sympatric speciation - the evolution of reproductive isolation between codistributed conspecific individuals - in generating biodiversity is highly controversial. Allochrony, or differences in breeding time (phenology) between conspecific individuals, has the potential to lead to reproductive isolation and therefore speciation. We critically review the literature to test the importance of allochronic speciation over the three timescales over which allochrony can occur - over the day, between seasons or between years - and explore what is known about genomic mechanisms underlying allochrony in the diverse taxa in which it is found. We found that allochrony can be a key contributor to reproductive isolation, especially if populations have little overlap in breeding time and therefore little potential for gene flow, and may sometimes be the initial or key driver of speciation. Shifts in phenology can be caused by several factors, including a new ecological opportunity, environmental change, or reinforcement. The underlying genomic basis of allochrony has been studied mostly in insects, highlighting the need for genomic studies in other taxa; nonetheless, results to date indicate that several cases of allochrony involve changes in circadian genes. This review provides the first comprehensive discussion of the role of allochrony in speciation and demonstrates that allochrony as a contributor to divergence may be more widespread than previously thought. Understanding genomic changes and adaptations allowing organisms to breed at new times may be key in the light of phenological changes required under climate change.


Fail-Safe System against Potential Tumorigenicity after Transplantation of iPSC Derivatives.
Human induced pluripotent stem cells (iPSCs) are promising in regenerative medicine. However, the risks of teratoma formation and the overgrowth of the transplanted cells continue to be major hurdles that must be overcome. Here, we examined the efficacy of the inducible caspase-9 (iCaspase9) gene as a fail-safe against undesired tumorigenic transformation of iPSC-derived somatic cells. We used a lentiviral vector to transduce iCaspase9 into two iPSC lines and assessed its efficacy in vitro and in vivo. In vitro, the iCaspase9 system induced apoptosis in approximately 95% of both iPSCs and iPSC-derived neural stem/progenitor cells (iPSC-NS/PCs). To determine in vivo function, we transplanted iPSC-NS/PCs into the injured spinal cord of NOD/SCID mice. All transplanted cells whose mass effect was hindering motor function recovery were ablated upon transduction of iCaspase9. Our results suggest that the iCaspase9 system may serve as an important countermeasure against post-transplantation adverse events in stem cell transplant therapies.


Heralded Spectroscopy Reveals Exciton-Exciton Correlations in Single Colloidal Quantum Dots.
Multiply excited states in semiconductor quantum dots feature intriguing physics and play a crucial role in nanocrystal-based technologies. While photoluminescence provides a natural probe to investigate these states, room-temperature single-particle spectroscopy of their emission has proved elusive due to the temporal and spectral overlap with emission from the singly excited and charged states. Here, we introduce biexciton heralded spectroscopy enabled by a single-photon avalanche diode array based spectrometer. This allows us to directly observe biexciton-exciton emission cascades and measure the biexciton binding energy of single quantum dots at room temperature, even though it is well below the scale of thermal broadening and spectral diffusion. Furthermore, we uncover correlations hitherto masked in ensembles of the biexciton binding energy with both charge-carrier confinement and fluctuations of the local electrostatic potential. Heralded spectroscopy has the potential of greatly extending our understanding of charge-carrier dynamics in multielectron systems and of parallelization of quantum optics protocols.


Detecting dissimulation: profiles of simulated malingerers, traumatic brain-injury patients, and normal controls on a revised version of Hiscock and Hiscock's Forced-Choice Memory Test.
A computer-administered memory test was given to normal subjects instructed to feign brain damage, normal controls, and traumatic brain-injured (TBI) patients with complaints of memory dysfunction. The test, a revised version of an instrument developed by Hiscock and Hiscock (1989) employed forced, two-choice recognition of previously presented five-digit numbers. Two levels of item difficulty and three retention intervals were used. Control and TBI subjects performed at near ceiling level on easy items and items with short retention intervals. Feigning subjects performed worse than TBI and control subjects at all levels of item difficulty and all retention intervals. Subjects groups were maximally distinguishable from one another by performance on difficult items and items with the longest retention interval. All TBI patients performed at or above chance level. Only 15% of the feigning subjects performed below chance level on any section of the test. Jack-knifed discriminant function analysis correctly classified 83% of all subjects into their respective groups. Although evaluation of patient performance relative to chance probability is useful for indicating the presence of extremely exaggerated memory deficits, criteria derived from sample distributions of group scores were superior for evaluation of motivation in less obvious cases. The Victoria Revision may be useful for detecting true memory deficit as well as dissimulation.


Reporting, appraising, and integrating data on genotype prevalence and gene-disease associations.
The recent completion of the first draft of the human genome sequence and advances in technologies for genomic analysis are generating tremendous opportunities for epidemiologic studies to evaluate the role of genetic variants in human disease. Many methodological issues apply to the investigation of variation in the frequency of allelic variants of human genes, of the possibility that these influence disease risk, and of assessment of the magnitude of the associated risk. Based on a Human Genome Epidemiology workshop, a checklist for reporting and appraising studies of genotype prevalence and studies of gene-disease associations was developed. This focuses on selection of study subjects, analytic validity of genotyping, population stratification, and statistical issues. Use of the checklist should facilitate the integration of evidence from these studies. The relation between the checklist and grading schemes that have been proposed for the evaluation of observational studies is discussed. Although the limitations of grading schemes are recognized, a robust approach is proposed. Other issues in the synthesis of evidence that are particularly relevant to studies of genotype prevalence and gene-disease association are discussed, notably identification of studies, publication bias, criteria for causal inference, and the appropriateness of quantitative synthesis.


What is the good of health care?
This paper sets out to discuss what precisely is meant by "benefit" when we talk of the requirement that the health care system concern itself with health gain or with maximising beneficial health care. In particular I argue that in discharging the duty to do what is most beneficial we need to choose between rival conceptions of what is meant by beneficial. One is the patient's conception of benefit and the second is the provider's or funder's conception of benefit. I argue that it is the patient's conception of benefit which is paramount and that if this is followed it commits us to a conception of patient care which must be blind to prognosis in so far as prognosis is thought to bear upon issues of prioritisation or resource allocation.


Fluid-filled blood pressure measurement systems.
The performance of catheter-manometer systems for the measurement of pulsatile pressure has been evaluated by both experimental techniques and theoretical considerations. The former approach has shown, on occasion, multiple maxima in the amplitude response. The latter has been approached in a variety of ways, ranging from extreme lumping to application of transmission line theory while employing different configurations in the system's representation. Multiple maxima have also been seen, The present paper identifies the sources of the differences found and compares the relative merits of various theoretical approaches. It introduces the compliance of the system as a figure of merit and provides a simple first-order approximation formula for evaluation of the quality of a system. Damping and impedance matching to improve the system's frequency response were studied. It was found that they were not needed in a very stiff or a very compliant system, nor should one worry about the representation of such a system.


Quazepam and flurazepam: differential pharmacokinetic and pharmacodynamic characteristics.
Quazepam and flurazepam share pharmacokinetic properties that result in prevention of early-morning insomnia, daytime rebound anxiety, and withdrawal rebound insomnia. Yet sleep laboratory and performance studies demonstrated that during a 1- to 4-week administration period quazepam had a low potential for causing daytime drowsiness or impairment. This profile may be related to several factors, such as differences in quazepam's metabolic pathways; plasma pharmacokinetics; rate of brain uptake, redistribution, and clearance; as well as differences in receptor binding and kinetics.


A High-Valent Non-Heme μ-Oxo Manganese(IV) Dimer Generated from a Thiolate-Bound Manganese(II) Complex and Dioxygen.
This study deals with the unprecedented reactivity of dinuclear non-heme MnII -thiolate complexes with O2 , which dependent on the protonation state of the initial MnII dimer selectively generates either a di-μ-oxo or μ-oxo-μ-hydroxo MnIV complex. Both dimers have been characterized by different techniques including single-crystal X-ray diffraction and mass spectrometry. Oxygenation reactions carried out with labeled 18 O2 unambiguously show that the oxygen atoms present in the MnIV dimers originate from O2 . Based on experimental observations and DFT calculations, evidence is provided that these MnIV species comproportionate with a MnII precursor to yield μ-oxo and/or μ-hydroxo MnIII dimers. Our work highlights the delicate balance of reaction conditions to control the synthesis of non-heme high-valent μ-oxo and μ-hydroxo Mn species from MnII precursors and O2 .


Open versus laparoscopic gastrojejunostomy for palliation in advanced pancreatic cancer.
To assess short-term outcome of open (OGJ) versus laparoscopic (LGJ) gastrojejunostomy in palliation of gastric outlet obstruction (GOO) caused by advanced pancreatic cancer, 22 OGJ patients were compared with 9 diagnosis-matched LGJ controls operated on at the same hospital between 1991 and 1996. Patients undergoing OGJ and LGJ were comparable for age, gender, weight, American Society of Anesthesiologists grading, and previous extensive abdominal surgery, but not for gastroenterostomy performed as a prophylactic procedure (9 vs. 0, respectively). Mortality (5 vs. 1, p = 1.5), overall morbidity (9 vs. 3, p = 0.42), operating time (113.6 +/- 24.5 minutes vs. 125 +/- 15.2 minutes, p < 0.5), time to oral solid food intake (7.2 +/- 0.9 days vs. 5.3 +/- 1.3 days, p < 0.5), nonsteroidal anti-inflammatory drug consumption (7,563.6 +/- 3,381.3 mg vs. 2,044 +/- 673 mg, p < 0.5), opioid consumption (688.5 +/- 258.6 mg vs. 2,910.5 +/- 2,659.9 mg, p < 0.5), delayed-return gastric emptying (5 vs. 1, p = 0.12), postoperative hospital stay (14.6 +/- 1.9 days vs. 10.1 +/- 1.8 days, p < 0.5), survival (5.7 +/- 0.8 months vs. 4.6 +/- 0.6 months, p < 0.5), and further hospital stay before death (9.8 +/- 3.3 days vs. 11.6 +/- 3.4 days, p > 0.5) were not significantly different in 22 OGJ and 9 LGJ patients, respectively. Estimated blood loss was significantly lower in LGJ patients (270.2 +/- 45.8 ml vs. 66 +/- 15.7 ml, p < 0.01). When 13 of 22 patients undergoing OGJ for treatment were compared with 9 LGJ patients, only estimated blood loss (p < 0.01) and hospital stay (p < 0.05) were significantly reduced in LGJ patients. Recurrent GOO before death occurred in one patient (1 of 22, 4.5%) 9 months after OGJ. LGJ for palliative treatment of GOO in advanced pancreatic cancer offered (in spite of the learning curve) reduced estimated blood loss and hospital stay when compared with OGJ.


Estimated cost-effectiveness of point-of-care testing in community pharmacies vs. self-testing and standard laboratory testing for HIV.
Point-of-care-testing (POCT) for HIV at community pharmacies can enhance care linkage compared with self-tests and increase testing uptake relative to standard lab testing. While the higher test uptake may increase testing costs, timely diagnosis and treatment can reduce downstream HIV treatment costs and improve health outcomes. This study provides the first evidence on the cost-effectiveness of pharmacist-led POCT vs. HIV self-testing and standard lab testing. Dynamic transmission model. We compared three HIV testing strategies: POCT at community pharmacies; self-testing using HIV self-test kits; and standard lab testing. Analyses were conducted from the Canadian health system perspective over a 30-year time horizon for all individuals aged 15-64 years in Canada. Costs were measured in 2021 Canadian dollars and effectiveness was captured using quality-adjusted life-years (QALYs). Compared with standard lab testing, POCT at community pharmacies would save $885 million in testing costs over 30 years. Though antiretroviral treatment costs would increase by $190 million with POCT as more persons living with HIV are identified and treated, these additional costs would be partly offset by their lower downstream healthcare utilization (savings of $150 million). POCT at community pharmacies would also yield over 5000 additional QALYs. Compared with HIV self-testing, POCT at community pharmacies would generate both higher costs and higher QALYs and would be cost-effective with an incremental cost-effectiveness ratio of $47 475 per QALY gained. Offering POCT at community pharmacies can generate substantial cost savings and improve health outcomes compared with standard lab testing. It would also be cost-effective vs. HIV self-testing.


Satisfaction and seizure outcomes of epilepsy surgery in tuberous sclerosis: A Swedish population-based long-term follow-up study.
We conducted a cross-sectional study to evaluate long-term outcomes of epilepsy surgery in tuberous sclerosis complex (TSC) in a Swedish population. Demographic and seizure data was retrieved from the Swedish National Epilepsy Surgery Registry and medical records. Patient reported outcome measurements (PROM) were determined by telephonic interviews at long term follow-up. Median follow-up was 6 y 8 m (range, 3-15 y 1 m) for tuberectomies (n = 15) and 3 y 6 m (range 2-10 y) for callosotomies (n = 7). Eight of the 15 tuberectomy participants were seizure-free. Four out of seven callosotomies were free from drop attacks. PROMs were provided by caregivers of 18/20 participants (data missing for two callosotomies). In the tuberectomy group, 6/8 patients were seizure-free and 3/7 had continued seizures; surgery was considered satisfactory and beneficial. Overall, satisfaction was high, even among patients who did not achieve remission; 13/15 tuberectomy responders recommended surgery to others with TSC and refractory epilepsy. None of the patients considered the surgery harmful. In the callosotomy group, satisfaction was low and congruent with the seizure outcome. All patients with continued drop attacks were unsatisfied; one considered surgery to be harmful. One participant, who would not recommend surgery to others, still perceived the surgery to be beneficial. This study confirmed that both tuberectomy and callosotomy are effective treatment options for TSC. Factors other than seizure outcomes seemed to have a major influence on satisfaction and perception of the benefit of surgery.


Paraneoplastic syndromes in urologic malignancy: the many faces of renal cell carcinoma.
Renal cell carcinoma is unique among the genitourinary malignancies in that close to one third of affected patients show signs and symptoms of a paraneoplastic syndrome. The paraneoplastic syndromes associated with renal cell carcinoma range from those manifesting in constitutional symptoms (ie, fever, cachexia, and weight loss) to those that result in specific metabolic and biochemical abnormalities (ie, hypercalcemia, nonmetastatic hepatic dysfunction, amyloidosis, etc). The presence of a paraneoplastic syndrome in a patient with renal cell carcinoma is neither a marker of metastatic disease nor necessarily indicative of a poor prognosis. The importance of understanding the pathophysiology and biology behind the many paraneoplastic syndromes associated with renal cell carcinoma lies in the fact that the presence of these protean symptoms may be the initial presentation of either primary or recurrent disease. In this review, we will describe the proposed mechanisms of action of the many paraneoplastic syndromes associated with renal cell carcinoma as well as outline the clinical evaluation and treatment options currently available for these noteworthy disorders.


Methylene Blue (Tetramethylthionine Chloride) Influences the Mobility of Adult Neural Stem Cells: A Potentially Novel Therapeutic Mechanism of a Therapeutic Approach in the Treatment of Alzheimer's Disease.
An interest in neurogenesis in the adult human brain as a relevant and targetable process has emerged as a potential treatment option for Alzheimer's disease and other neurodegenerative conditions. The aim of this study was to investigate the effects of tetramethylthionine chloride (methylene blue, MB) on properties of adult murine neural stem cells. Based on recent clinical studies, MB has increasingly been discussed as a potential treatment for Alzheimer's disease. While no differences in the proliferative capacity were identified, a general potential of MB in modulating the migratory capacity of adult neural stem cells was indicated in a cell mobility assay. To our knowledge, this is the first time that MB could be associated with neural mobility. The results of this study add insight to the spectrum of features of MB within the central nervous system and may be helpful for understanding the molecular mechanisms underlying a potential therapeutic effect of MB.


Sexual behaviour and contraceptive use among secondary school students in Ilesha south west Nigeria.
Data from several parts of Nigeria point to increasing sexual activity among single adolescents of both sexes with progressive decreasing age at initiation and poor contraceptive use. This paper investigates the sexual behaviour and contraceptive use among teenage secondary school students in Ilesha, southwest Nigeria. This is a cross-sectional population survey of 300 male and female secondary school students within the age group of 13-19 years. The setting is secondary school coaching classes in Ilesha, southwest Nigeria. The outcome measures include prevalence of sexual activity, age at first sexual debut, circumstances leading to first sexual debut, number of sexual partners and family planning use. The result shows that out of the 300 studied, 50% were sexually active, the predominant age at first coitus was 15-19 years, and circumstances leading to sexual debut included mutual agreement, coercion and curiosity. Predominant proportion of sexually active teenagers (86.7%) did not use contraception at the time of first coitus and most of them had more than one sexual partner. The conclusion from this study is that 50% of teenage secondary school girls in this part of Nigeria are sexually active; 68.7% whom, have multiple sexual partners, and 86.7% of them did not use contraception at sexual debut. This unsafe sexual behaviour therefore put them at a great risk of acquiring STDs including HIV infection, and unwanted pregnancy. This study therefore recommends sex education/family life education for young people to encourage them to delay sexual activity as much as possible and practice safe sex when it eventually commences. There is also the need to sensitise the young people, parents, teachers, the community and all stake holders on the magnitude of the problem and to open up dialogue that will break the social, cultural and other mysteries hindering adolescents and youth reproductive health education and services in Nigeria.


High standards. A decade after the law went into effect, there is still debate about the pros and cons of the HIPAA privacy and electronic transaction regulations.
When congress passed the Health Insurance Portability and Accountability act in 1996, the goal was to create a simpler, more standardized system that would eventually lower health care costs; reduce errors through safe, universally accepted electronic communication of health care transactions; and eliminate paper claims. Ten years later, the jury is still out on whether HIPAA has been worth the time, energy, and financial investment for insurers. That's not to say, however, that HIPAA hasn't generated benefits while also creating new challenges. "Standards made sense," says Tom Fitzpatrick, Horizon Blue Cross Blue Shield of New Jersey's director of enterprise strategic planning, "but no one ever said it would be fast, cheap, or easy. It was challenging to integrate proprietary claims systems and legacy software with the new standards that took effect in October 2003. But that wasn't the end of the story. HIPAA's privacy and security rules and the standard identifiers have meant even more upgrades and improvements and have required payers to spend millions of additional dollars over the past three years on HIPAA compliance." According to a set of quarterly surveys conducted by HIMSS/Phoenix Health Systems, progress has actually been fairly rapid. On the other hand, some things have remained much the same. In 2003, payers cited "understanding/interpreting the legal requirements" as the most difficult aspect of the HIPAA remediation process, followed by "achieving successful integration of new policies and procedures" and "resolving issues with third parties". In 2006, the barriers are similar, with users citing the same top two struggles.


Is the first urinary infection in childhood a definite indication for radiological examination?
The need for X-ray investigations of the urinary tract at the time of the first proven urinary infection, even in older girls, is stressed. This policy is the best way to detect chronic pyelonephritis and congenital anomalies of the urinary tract and provide timely and appropriate treatment. These conclusions are supported by personal experience (see case description), and the incidence of congenital anomalies encountered during an 11 year period in our children's hospital. There were 957 concecutive cases with X-ray studies of the urinary tract (IVP and MCU). A considerable proportion (30%) were older children (4-12 years) having supravesical obstruction defined when the X-ray studies were performed at the first urinary infection. The proportion of abnormalities rose to 66% when the investigations were done after recurrent infections, which may contribute to the development and progression of disease.


Screening for congenital neural tube defects in a high-risk area: an epidemiological perspective.
Data from the Glasgow Register of Congenital Malformations were used to investigate the extent of the recent decline in the prevalence of anencephaly and spina bifida, and the contribution of antenatal screening to it. Over the period 1974-85 inclusive, 303 pregnancies with an anencephalic foetus were diagnosed, representing an "adjusted" prevalence of 1.9 per 1000 total births, of which 179 (59%) were terminated following antenatal screening. There were 364 pregnancies with a spina bifida foetus representing an "adjusted" prevalence of 2.3 per 1000 total births, of which 84 (23%) were terminated. Over the study period, the "adjusted" prevalence of anencephaly fell by 50% while the birth prevalence fell by 89%; the "adjusted" prevalence of spina bifida fell by 38% while the birth prevalence fell by 76%. It was concluded that although the birth prevalence of both defects (particularly anencephaly) would have declined substantially in the absence of screening, the West of Scotland programme should continue.


Needs assessment survey for simulation-based training for gastrointestinal endoscopy nurses.
The optimal performance of gastrointestinal (GI) endoscopy nurses is required for patient safety and quality improvement. The aim of the present study was to assess the educational needs for simulation-based training for Korean GI endoscopy nurses using importance-performance analysis. A cross-sectional survey was conducted with 238 Korean nurses from 25 endoscopy units. The educational needs of these nurses were identified using the 35 item clinical competence importance-performance scale. Exploratory factor analysis of the scale identified the following eight factors: emergency care, patient monitoring, evidence-based practice, documentation and referral, patient safety, nursing process, patient assessment, and infection control. A significant overall mean difference was identified between importance and performance for all eight factors, with emergency care showing the largest difference. It was also ranked the highest priority for continuing education in the importance-performance analysis matrix. Therefore, simulation-based training should focus on enhancing emergency care competence for GI endoscopy nurses to improve patient safety and quality of care.


OrthoPilot total hip arthroplasty workflow and surgery.
To analyze the workflow and navigation principles of the OrthoPilot total hip arthroplasty (THA) Version 2.0 (B. Braun-Aesculap, Tuttlingen, Germany), a consecutive series of 60 patients was used to compare the navigated intraoperative data and postoperative measurements of stem and cup position. Within the safe zone, 96.3% of 54 acetabular cups were positioned. The rotational accuracy of femoral instruments was 65 degrees in 41 THAs. The femoral offset was medialized by 6.05 mm in 76% and lateralized by 2.1 mm in 14%. The data for leg length and range of motion from 60 THAs and the navigated data were similar. Thus, the first clinical validation of the workflow of the OrthoPilot THA Version 2.0 is encouraging.


Potent acyl-CoA synthetase 10 inhibitors kill Plasmodium falciparum by disrupting triglyceride formation.
Identifying how small molecules act to kill malaria parasites can lead to new "chemically validated" targets. By pressuring Plasmodium falciparum asexual blood stage parasites with three novel structurally-unrelated antimalarial compounds (MMV665924, MMV019719 and MMV897615), and performing whole-genome sequence analysis on resistant parasite lines, we identify multiple mutations in the P. falciparum acyl-CoA synthetase (ACS) genes PfACS10 (PF3D7_0525100, M300I, A268D/V, F427L) and PfACS11 (PF3D7_1238800, F387V, D648Y, and E668K). Allelic replacement and thermal proteome profiling validates PfACS10 as a target of these compounds. We demonstrate that this protein is essential for parasite growth by conditional knockdown and observe increased compound susceptibility upon reduced expression. Inhibition of PfACS10 leads to a reduction in triacylglycerols and a buildup of its lipid precursors, providing key insights into its function. Analysis of the PfACS11 gene and its mutations point to a role in mediating resistance via decreased protein stability.


Total mercury and methylmercury in high altitude surface snow from the French Alps.
Surface snow samples were collected weekly from the 31st of December 2008 to the 21st of June 2009 from Lake Bramant in the French Alps. Total mercury (THg), total dissolved mercury (THgD), methylmercury (MeHg) and particle distributions in surface snow were analyzed. Results showed that THg concentrations, MeHg concentrations and particle load increased with snow surface temperature, which is an indicator of rising temperatures as the season progresses. Significant correlations between MeHg and snow surface temperature and MeHg and total particles greater than 10 μm were observed. This suggests that the MeHg found in the snow originates from atmospheric deposition processes rather than in situ snowpack sources. This study suggests that an important post-winter atmospheric deposition of MeHg and THg occurs on summital zones of the French Alps and it is likely that this contamination originates from the surrounding valleys.


[Developing and validating of patient-reported questionnaires - part 1].
Patient-reported outcome measurements (PROMs) are often questionnaires which provide and rate the patient's point of view in the measurement of subjective clinical phenomena such as pain or quality of life. In the first of two articles we describe five of the six developmental phases of a patient-reported outcome questionnaire. Adherence to good methodology throughout the process is of utmost importance to ensure reliable and valid results from studies using PROM. Patients from the target population should be involved in the process to obtain content and face validation, i.e. through focus groups.


[Thyroid hormone dynamics during the thyrostatic treatment of Basedow's disease].
The efficacy of the generally accepted therapeutic scheme was studied in 52 newly discovered and untreated patients with Basedow's disease with the purpose to clarify the frequent failures of the thyrostatic treatment. The thyroid hormones levels were determined in dynamics initially and in the course of the disease. The results show that the efficacy of the thyrostatic drugs is individually different and depends on the supersensitivity of the thyroid gland. The remission occurred on the average on the 50th day--considerably earlier than the usually expected time limit. In order to study the expedience of the therapeutic scheme applied, the active dose required by the scheme was continued. As a result in the 4 month after the beginning of the treatment the thyrotropic hormone increased to a mean level of 35.7 mU/l--hypothyroid value. A monthly control of the thyroid hormones levels during the course of the active treatment is necessary and a reduction of the doses of the thyrostatic drugs when a remission is achieved must be performed.


Increased sodium conductance in the synaptic region of rat skeletal muscle fibres.
Differences in sodium conductance between end-plate and extrajunctional regions of rat lumbrical muscle fibres were measured by comparing action potential maximum rate of rise (Vmax) in the two regions and by using a vibrating micro-electrode to record steady inward current produced by application of veratridine. In normal Krebs solution, action potential Vmax was significantly greater (by 43%) in the end-plate region than in extrajunctional regions of the fibres. When chloride conductance was greatly reduced by bathing muscles in solutions with low chloride concentration, Vmax was still significantly higher (by 28%) in the end-plate region than in extrajunctional regions. The increased Vmax could be recorded only within a distance of about 150-200 microns of the end-plate. Steady inward current was recorded with a vibrating micro-electrode at the end-plate in response to veratridine; the current persisted when veratridine was introduced in low-chloride Krebs solution, and it was rapidly reversed by tetrodotoxin. The current reflected a 5 mV difference in membrane potential between the end-plate region and extrajunctional regions. The results suggest that sodium conductance is increased in the synaptic region relative to extrajunctional regions of the fibres.


Hidden in plain sight: a morphological study revealing three new species of the skipper genus Drephalys Watson, 1893 (Lepidoptera: Hesperiidae) from Brazil.
Three new species of Drephalys Watson, 1893 from Brazil are described as follows: D. dracarys Madruga, Siewert, Mielke Dolibaina, sp. n. from Acre, Mato Grosso and Rondônia states, D. electrinus Siewert, Madruga, Mielke Dolibaina, sp. n. from Acre and Amazonas states, and D. citrinus Madruga, Siewert, Mielke Casagrande, sp. n. from Amazonas state. Drephalys heraclides Bell, 1942 was reported for the first time from Brazil. To characterize the new species, illustrations of the male genitalia of D. heraclides, D. phoenice (Hewitson, 1867) and D. phoenicoides (Mabille Boullet, 1919) are provided. Diagnosis, adult photos, illustration of the male genitalia and a geographical distribution map are provided for the new species.


The medical care programs of the Farm Security Administration, 1932 through 1947: a rehearsal for national health insurance?
At a time of renewed interest in universal health insurance, an examination of earlier periods when society grappled with the link between socioeconomic status and health is fruitful. Between 1935 and 1947, the federal government sponsored a comprehensive medical care program for low-income farmers, sharecroppers, and migrant workers under the auspices of the Farm Security Administration (FSA). Despite the strong opposition of the American Medical Association, humanitarian and economic concerns at the local level often promoted physicians' participation in the program's group prepayment plans. Many FSA leaders clearly saw the program as a model upon which national health insurance might advance. However, in the wake of World War II, the FSA program declined as physicians' income improved, the rural population declined, and traditional ideological objections to federal intervention in medical care resurfaced. The FSA experience illuminates the complex ideological, economic, and humanitarian motivations of American physicians in the face of health care reform.


Cooperative problem solving with personal mobile information tools in hospitals.
Health-care professionals have a broad range of needs for information and cooperation while working at different points of care (e.g., outpatient departments, wards, and functional units such as operating theaters). Patient-related data and medical knowledge have to be widely available to support high-quality patient care. Furthermore, due to the increased specialization of health-care professionals, efficient collaboration is required. Personal mobile information tools have a considerable potential to realize almost ubiquitous information and collaborative support. They enable to unite the functionality of conventional tools such as paper forms, dictating machines, and pagers into one tool. Moreover, they can extend the support already provided by clinical workstations. An approach is described for the integration of mobile information tools with heterogeneous hospital information systems. This approach includes identification of functions which should be provided on mobile tools. Major functions are the presentation of medical records and reports, electronic mailing to support interpersonal communication, and the provision of editors for structured clinical documentation. To realize those functions on mobile tools, we propose a document-based client-server architecture that enables mobile information tools to interoperate with existing computer-based application systems. Open application systems and powerful, partially wireless, hospital-wide networks are the prerequisites for the introduction of mobile information tools.


Triclosan antimicrobial polymers.
Triclosan antimicrobial molecular fluctuating energies of nonbonding electron pairs for the oxygen atom by ether bond rotations are reviewed with conformational computational chemistry analyses. Subsequent understanding of triclosan alternating ether bond rotations is able to help explain several material properties in Polymer Science. Unique bond rotation entanglements between triclosan and the polymer chains increase both the mechanical properties of polymer toughness and strength that are enhanced even better through secondary bonding relationships. Further, polymer blend compatibilization is considered due to similar molecular relationships and polarities. With compatibilization of triclosan in polymers a more uniform stability for nonpolar triclosan in the polymer solid state is retained by the antimicrobial for extremely low release with minimum solubility into aqueous solution. As a result, triclosan is projected for long extended lifetimes as an antimicrobial polymer additive. Further, triclosan rapid alternating ether bond rotations disrupt secondary bonding between chain monomers in the resin state to reduce viscosity and enhance polymer blending. Thus, triclosan is considered for a polymer additive with multiple properties to be an antimicrobial with additional benefits as a nonpolar toughening agent and a hydrophobic wetting agent. The triclosan material relationships with alternating ether bond rotations are described through a complete different form of medium by comparisons with known antimicrobial properties that upset bacterial cell membranes through rapid fluctuating mechanomolecular energies. Also, triclosan bond entanglements with secondary bonding can produce structural defects in weak bacterial lipid membranes requiring pliability that can then interfere with cell division. Regarding applications with polymers, triclosan can be incorporated by mixing into a resin system before cure, melt mixed with thermoplastic polymers that set on cooling into a solid or alternatively applied as a coating through several different methods with dissolving into an organic solvent and dried on by evaporation as a common means.


Dissociation of the effects of gonadal steroids on brain serotonin metabolism and sexual behavior in the male rat.
Steady-state concentrations of 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in anterior hypothalamus (AH), preoptic area (POA), suprachiasmatic nucleus (SCN) and ventromedial nucleus (VMH) were equivalent 30 days after castration or sham operation in male rats killed at either the middle of the light or dark periods of a 12 : 12 h day/night cycle. Chronic administration of testosterone, 5alpha-dyhydrotestosterone (DHT), estradiol-17beta (E2) or E2+DHT for 30 days to castrated males also failed to alter differentially the concentrations of 5-HT and 5-HIAA in any of the brain regions assayed. Although castrated and sham-operated rats had similar levels of 5-HT and 5-HIAA in whole hypothalamus + preoptic area (H + POA), brain stem (BS), and cortex (CTX) when measured 7 and 32 days postoperatively, the mounting rates and the percentage of animals ejaculating were similar for both groups 6 days postoperatively whereas only the sham-operated animals continued to mount at a high rate and to ejaculate 31 days postoperatively. Also, 30 days after surgery, there were no differences among groups of males which were either castrated, castrated and given a subcutaneous capsule of testosterone, or sham operated, in the accumulation of 5-hydroxytryptophan in the H + POA, BS, or CTX measured 30 min after an intraperitoneal injection of an aromatic amino acid decarboxylase inhibitor. These results suggest the the hormonal activation of masculine sexual behavior in the rat is not mediated by a change in the metabolic activity of 5-HY neurons in either the hypothalamus or POA.


Whole genome sequencing of an unusual serotype of Shiga toxin-producing Escherichia coli.
Shiga toxin-producing Escherichia coli serotype O117:K1:H7 is a cause of persistent diarrhea in travelers to tropical locations. Whole genome sequencing identified genetic mechanisms involved in the pathoadaptive phenotype. Sequencing also identified toxin and putative adherence genes flanked by sequences indicating horizontal gene transfer from Shigella dysenteriae and Salmonella spp., respectively.