Clinical features of 2041 human brucellosis cases in China.
Human brucellosis has become a major public health problem in China. However, the available clinical data on brucellosis cases are limited. We retrospectively reviewed medical charts of 2041 patients with confirmed human brucellosis and prospectively recorded their outcomes by telephone interview. These patients were admitted to the Sixth People's Hospital of the Xinjiang Uygur Autonomous region between 1st January and 31st December 2014. Data on these patients were collected from hospital medical records. Many patients presented with fatigue (67%), fever (64%), arthralgia (63%) and sweating (54%). High erythrocyte sedimentation rate (ESR) (69%), high C-reactive protein (CRP) (39%), high alanine aminotransferase (ALT) (33%) and high aspartate aminotransferase (AST) (20%) were the most common laboratory findings, especially in acute patients. There was evidence of focal involvement in 90% of patients. A total of 61.5% of brucellosis patients recovered. Multivariate logistic regression analyses suggested that the risk factors key to unfavorable prognosis were: age≥45 years (OR = 1.75, 95% CI 1.36-2.24), back pain (OR = 1.50, 95% CI 1.16-1.94) and joint tenderness (OR = 1.73, 95% CI 1.13-2.65). The increasing duration of the illness increased the risk of poor prognosis. Patients with brucellosis showed different characteristics in different clinical stages. In China, the chronicity rate of human brucellosis is high. The risk of poor prognosis is increased in patients aged 45 years or older, patients who have had brucellosis for a substantial period of time, and patients with back pain or joint tenderness. The clinical management of brucellosis should be improved to include sensitive diagnostic methods for subacute and chronic brucellosis.


Prestroke physical activity could influence acute stroke severity (part of PAPSIGOT).
To investigate the influence of prestroke physical activity (PA) on acute stroke severity. Data from patients with first stroke were retrieved from registries with a cross-sectional design. The variables were PA, age, sex, smoking, diabetes, hypertension and statin treatment, stroke severity, myocardial infarction, new stroke during hospital stay, and duration of inpatient care at stroke unit. PA was assessed with Saltin-Grimby's 4-level Physical Activity Level Scale, and stroke severity was assessed with the National Institutes of Health Stroke Scale (NIHSS). Logistic regression was used to predict stroke severity, and negative binomial regression was used to compare the level of PA and stroke severity. The study included 925 patients with a mean age of 73.1 years, and 45.2% were women. Patients who reported light or moderate PA levels were more likely to present a mild stroke (NIHSS score 0 to 5) compared with physically inactive patients in a model that also included younger age as a predictor (odds ratio = 2.02 for PA and odds ratio = 0.97 for age). The explanatory value was limited at 6.8%. Prestroke PA was associated with less severe stroke, and both light PA such as walking at least 4 h/wk and moderate PA 2-3 h/wk appear to be beneficial. Physical inactivity was associated with increased stroke severity. This study suggests that PA and younger age could result in a less severe stroke. Both light PA such as walking at least 4 h/wk and moderate PA 2-3 h/wk appear to be beneficial.


Mutagenic and DNA repair activity in traffic policemen: a case-crossover study.
Emissions from vehicles are composed of heterogeneous mixtures of hazardous substances; several pollutants such as Polycyclic Aromatic Hydrocarbons (PAHs) are amongst the most dangerous substances detected in urban monitoring. A cohort of traffic policemen usually occupationally exposed to PAHs present in the urban environment were examined in order to assess the mutagenicity and DNA capacity repair. Seventy-two urban traffic policemen working in Catania's metropolitan area were enrolled in the study. Two spot urine samples were collected from each subject during the whole working cycle as follows: sample 1 (S1), pre-shift on day 1; sample 2 (S2) post-shift on day 6. 1-hydroxypyrene (1-OHP) was measured to serve as an indirect exposure indicator. Urinary mutagenic activity was assessed through the plate incorporation pre-incubation technique with S9, using YG1024 Salmonella typhimurium strain over-sensitive to PAH metabolite. Concentrations of urinary 8-oxodG were measured using liquid chromatography tandem mass spectrometry. As regards the exposure to PAHs, results highlighted a statistically significant difference (p < 0.001) between pre-shift on day 1 and post-shift on day 6 levels. Mutagenic activity was detected in 38 (66%) workers on S1 and in 47 (81%) on S2. Also 8-oxodG analysis showed a statistically significant difference between S1 and S2 sampling. This study demonstrated that occupational exposure to pollutants from traffic emission, assessed via 1-OHP measurements in urine, may lead to DNA repair and mutagenic activity, in line with other studies.


Over-the-counter medications for acute cough in children and adults in ambulatory settings.
Acute cough due to upper respiratory tract infection (URTI) is a common symptom. Non-prescription over-the-counter (OTC) medicines are frequently recommended as a first-line treatment, but there is little evidence as to whether these drugs are effective. To assess the effects of oral OTC cough preparations for acute cough. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2006, Issue 4); MEDLINE (January 1966 to January Week 1, 2007); EMBASE (January 1974 to January 2007); and the UK Department of Health National Research Register (June 2007). Randomised controlled trials (RCTs) comparing oral OTC cough preparations with placebo in children and adults suffering from acute cough in ambulatory settings. We considered all cough outcomes and second outcomes of interest were adverse effects. Two review authors independently screened potentially relevant citations and independently extracted data and assessed study quality. Quantitative analysis was performed where appropriate. Twenty five trials (17 in adults, 8 in children) involving 3492 people (2876 adults and 616 children) were included. RESULTS OF STUDIES IN ADULTS: Six trials compared antitussives with placebo and had variable results. Two trials compared the expectorant, guaifenesin with placebo, one indicated significant benefit whereas the other did not. One trial found that a mucolytic reduced cough frequency and symptom scores. Two studies examined antihistamine-decongestant combinations and found conflicting results. Three studies compared other combinations of drugs with placebo and indicated some benefit in reducing cough symptoms. Three trials found antihistamines were no more effective than placebo in relieving cough symptoms. RESULTS OF STUDIES IN CHILDREN: Antitussives (two studies), antihistamines (two studies), antihistamine decongestants (two studies) and antitussive/bronchodilator combinations (one study) were no more effective than placebo. No studies using expectorants met our inclusion criteria. The results of one trial favoured active treatment with mucolytics over placebo. One trial tested two paediatric cough syrups and both preparations showed a 'satisfactory response' in 46% and 56% of children compared to 21% of children in the placebo group. There is no good evidence for or against the effectiveness of OTC medicines in acute cough. The results of this review have to be interpreted with caution due differences in study characteristics and quality. Studies often showed conflicting results with uncertainty regarding clinical relevance. Higher quality evidence is needed to determine the effectiveness of self-care treatments for acute cough.


[The problems of the work rehabilitation of breast cancer patients after radical treatment].
The paper discusses issues in occupational rehabilitation of breast cancer patients following radical treatment. A high level of labor orientation is emphasized which is to be considered in assessing working ability in an individual patient. As many as 71.2% of patients resumed working. Improvement in functional and psychoemotional status observed in the majority of patients as a result of rehabilitation treatment contributed to orientation to resume social activity.


Recombinant tissue plasminogen activator in the treatment of intraventricular hemorrhage secondary to periventricular arteriovenous malformation before surgery: case report.
Intraventricular hemorrhage (IVH) is known to cause acute obstructive hydrocephalus, refractory elevated intracranial pressures (ICPs), and lowered cerebral perfusion pressures, leading to cortical ischemia. Frequent obstruction of external ventricular drains as a result of thrombus is a recurring theme. We present a case of IVH secondary to periventricular arteriovenous malformation (AVM) that was not visible at admission angiography and was treated by intraventricular infusion of recombinant tissue plasminogen activator before surgical intervention. An 11-year-old boy presented with acute onset of headache followed by two seizures, loss of consciousness, decerebration, right temporal hematoma, IVH, and acute obstructive hydrocephalus. A right external ventricular drain was placed but functioned poorly. ICP could not be controlled by conventional methods. Five milligrams of recombinant tissue plasminogen activator was injected into the ventricular system via the external ventricular drain. This was repeated daily for 4 days. This treatment resulted in progressive improvement in ICP and clinical status. Once the clot partially cleared, magnetic resonance imaging and magnetic resonance angiography suggested the presence of a right periventricular arteriovenous malformation, which was confirmed by angiography and subsequently resected. Recombinant tissue plasminogen activator is effective in resolving IVH causing obstructive hydrocephalus and uncontrollable ICP posing a life-threatening situation, secondary to ruptured arteriovenous malformation, before surgical intervention.


Impact of the National Cancer Act on grant support.
The National Cancer Act of 1971 resulted in a threefold increase in appropriations for the National Cancer Institute (NCI) within a 4-year period. A major effect was the increase for the Grants Program from +93 million in fiscal year 1970 to more than +280 million in 1974. Grant programs, administered by the Division of Cancer Research Resources and Centers, account for more than 50% of the total NCI extramural research budget and fall into four broad categories: research, training (including fellowships), cancer control, and construction. With the exception of the training area, funding for all grant programs has increased dramatically as a result of the Act. The ocst of research has also risen, as reflected in the average twofold increase in cost per NCI traditional grant over the past 10 years. This rise in cost is due to a number of factors, including inflation, more sophisticated equipment and supplies and, in some cases, more ambitious projects. The principal type of research grants include traditional awarded for investigator-initiated research projects, and center, awarded for comprehensive and specialized cancer centers. While support for traditional grants has remained in the forefront of NCI funding, money for cancer center grants has increased at a greater rate in recent years, reflecting emphasis on the development of cancer centers throughout the country. Compared to other institutes at the NIH, NCI is in a very favorable funding position; in fiscal year 1974 NCI awarded more money for its research grant programs than all of the other institutes (with the exception of the National Heart and Lung Institue) obligated for their entire budgets. The Act has stimulated a large increase in new cancer applications received, and the increased funding has made it possible for NCI to award a greater number of grants. Young investigators have competed well for the additional monies made available by the Act and funding for cancer research outside the United States, still only a small part of NCI's budget, has increased.


Understanding Memories of the Past in the Context of Different Complex Neural Network Architectures.
Deep learning (primarily using backpropagation) and neuroevolution are the preeminent methods of optimizing artificial neural networks. However, they often create black boxes that are as hard to understand as the natural brains they seek to mimic. Previous work has identified an information-theoretic tool, referred to as R, which allows us to quantify and identify mental representations in artificial cognitive systems. The use of such measures has allowed us to make previous black boxes more transparent. Here we extend R to not only identify where complex computational systems store memory about their environment but also to differentiate between different time points in the past. We show how this extended measure can identify the location of memory related to past experiences in neural networks optimized by deep learning as well as a genetic algorithm.


CAGW Peptide- and PEG-Modified Gene Carrier for Selective Gene Delivery and Promotion of Angiogenesis in HUVECs in Vivo.
Gene therapy is a promising strategy for angiogenesis, but developing gene carriers with low cytotoxicity and high gene delivery efficiency in vivo is a key issue. In the present study, we synthesized the CAGW peptide- and poly(ethylene glycol) (PEG)-modified amphiphilic copolymers. CAGW peptide serves as a targeting ligand for endothelial cells (ECs). Different amounts of CAGW peptide were effectively conjugated to the amphiphilic copolymer via heterofunctional poly(ethylene glycol). These CAG- and PEG-modified copolymers could form nanoparticles (NPs) by self-assembly method and were used as gene carriers for the pEGFP-ZNF580 (pZNF580) plasmid. CAGW and PEG modification coordinately improved the hemocompatibility and cytocompatibility of NPs. The results of cellular uptake showed significantly enhanced internalization efficiency of pZNF580 after CAGW modification. Gene expression at mRNA and protein levels demonstrated that EC-targeted NPs possessed high gene delivery efficiency, especially the NPs with higher content of CAGW peptide (1.16 wt %). Furthermore, in vitro and in vivo vascularization assays also showed outstanding vascularization ability of human umbilical vein endothelial cells treated by the NP/pZNF580 complexes. This study demonstrates that the CAGW peptide-modified NP is a promising candidate for gene therapy in angiogenesis.


Simultaneous quantitative analysis of uremic toxins by LC-MS/MS with a reversed-phase/cation-exchange/anion-exchange tri-modal mixed-mode column.
Column choice is crucial to the development of liquid chromatography/tandem mass spectrometry (LC-MS/MS) methods because analyte selectivity is dependent on the nature of the stationary phase. Recently, mixed-mode chromatography, which employs a combination of two or more stationary phases and solvent systems, has emerged as an alternative to multiple, complementary, single-column systems. This report describes the development and validation of a novel analytical method based on LC-MS/MS employing a reversed-phase/cation-exchange/anion-exchange tri-modal column (Scherzo SS-C18; Imtakt) for the simultaneous quantification of various uremic toxins (UTx), including creatinine, 1-methyladenosine, trimethylamine-N-oxide, indoxyl sulfate, p-cresyl sulfate, phenyl sulfate and 4-ethylphenyl sulfate. Stable isotope-labeled compounds were prepared as internal standards (ISs) for each analyte. Mobile phase optimization and appropriate gradient conditions resulted in satisfactory retention and peak resolution that could not have been attained with a single stationary phase LC system. The essential validation parameters, including intra- and inter-assay precision and accuracy, were adequate. The validated method was applied to measure serum levels of the aforementioned compounds in 19 patients with chronic kidney disease. This is the first report detailing the simultaneous quantification of these analytes using stable isotopes as ISs. Our results suggest that Scherzo SS-C18 columns will be considered breakthrough tools in the development of analytical methods for compounds that are difficult to quantify simultaneously in traditional LC systems.


Absence of MSY2 in mouse oocytes perturbs oocyte growth and maturation, RNA stability, and the transcriptome.
Messenger RNA is remarkably stable during oocyte growth, thus enabling mRNAs to accumulate during the growth phase and thereby provide mRNAs that support early embryonic development. MSY2, a germ cell-specific RNA-binding protein, is implicated in regulating mRNA stability. MSY2 is essential for development because female Msy2(-/-) mice are infertile. We describe here the characterization of Msy2(-/-) oocytes. Mutant oocytes grow more slowly during the first wave of folliculogenesis, and maturation to and arrest at metaphase II is severely compromised because of aberrant spindle formation and chromosome congression. Consistent with MSY2 conferring mRNA stability is that the amount of poly(A)-containing RNA is reduced by ~25% in mutant oocytes. Stability of an exogenous mRNA injected into mutant oocytes is lower than when compared to their wild-type counterparts, and moreover, expression of wild-type MSY2 in mutant oocytes increases mRNA stability, whereas injection of a mutant form of MSY2 not capable of binding RNA does not. Transcription quiescence that normally occurs during the course of oocyte growth is not observed in mutant oocytes, and the transcriptome of mutant oocytes is markedly perturbed. These results, and those of previous studies, strongly implicate a central role of MSY2 in regulating mRNA stability.


Association of Stress Hyperglycemia Ratio With Acute Ischemic Stroke Outcomes Post-thrombolysis.
Background and Purpose: The association between stress hyperglycemia and clinical outcomes in patients with acute ischemic stroke treated with intravenous thrombolysis (IVT) is uncertain. We sought to analyze the association between the stress hyperglycemia ratio (SHR) using different definitions and clinical outcomes in acute patients with ischemic stroke undergoing IVT. Methods: A total of 341 patients with ischemic stroke receiving IVT were prospectively enrolled in this study. The SHR was evaluated using different equations: SHR1, fasting glucose (mmol/L)/glycated hemoglobin (HbA1c) (%); SHR2, fasting glucose (mmol/L)/[(1.59 × HbA1c)-2.59]; SHR3, admission blood glucose (mmol/L)/[(1.59 × HbA1c)-2.59]. A poor functional outcome was defined as a modified Rankin scale score of 3-6 at 3 months. Multivariate logistic regression analysis was used to identify the relationship between different SHRs and clinical outcomes after IVT. Results: A total of 127 (37.2%) patients presented with poor functional outcomes at 3 months. The predictive value of SHR1 for poor functional outcomes was better than that of SHR2 and SHR3 in receiver operating characteristic analyses. On multivariate analysis, SHR1 [odds ratio (OR) 14.639, 95% CI, 4.075-52.589; P = 0.000] and SHR2 (OR, 19.700; 95% CI; 4.475-86.722; P = 0.000) were independently associated with an increased risk of poor functional outcome but not SHR3. Conclusions: Our study confirmed that the SHR, as measured by SHR1 and SHR2, is independently associated with worse clinical outcomes in patients with ischemic stroke after intravenous thrombolysis. Furthermore, SHR1 has a better predictive performance for outcomes than other SHR definitions.


Genetic mapping in Methylophilus methylotrophus AS1.
Prime plasmids have been isolated from Methylophilus methylotrophus AS1 using the IncP-1 plasmid pMO172. Such plasmids can complement mutant function when transferred to appropriate strains of Pseudomonas aeruginosa PAO. From the range of particular functions complemented by each prime plasmid, a preliminary map of the M. methylotrophus AS1 genome with four groups of linked markers was obtained. Physical examination of two of these prime plasmids showed that each had acquired an additional DNA segment. Southern hybridization experiments showed there was sequence homology between the additional DNA segments and M. methylotrophus AS1 DNA, confirming that these prime plasmids carried a segment of the M. methylotrophus AS1 genome. Mapping by complementation may be applicable to other bacteria in which mutants suitable for selecting recombinants are not readily available.


Pharmacokinetics of radiolabeled polyclonal antiferritin in patients with Hodgkin's disease.
The objective was to identify pharmacokinetic parameters predictive for tumor response and normal tissue side effects after i.v. administered radiolabeled rabbit antihuman ferritin IgG. Twenty-eight patients with recurrent Hodgkin's disease received 2 mg of rabbit antihuman ferritin i.v., labeled with 4-7 mCi of In-111 followed by two doses of 0.25, one dose of 0.3, or one dose of 0.4 mCi of Y-90-labeled antiferritin per kg of body weight 1 week later. Radioactivity and HPLC measurements of blood and urine samples and liver and tumor volumes identified on sequential whole-body scans provided the data for a pharmacokinetic analysis covering the first 6 days after the administration of the radioimmunoconjugate. Side effects and tumor response were recorded. Temporary hematological toxicity was noted in all patients. Sixteen patients showed a tumor response. The Y-90 blood level at 1 h after administration correlated with the severity of subsequent hematological toxicity. The rapid blood elimination half-life of radioactivity was 4.4 h. Less than 5% of the administered radioactivity was eliminated in the first 24 h urine. The slow blood elimination half-life was 44 and 37 h for In-111 and Y-90, respectively. One of 12 retreated patients produced anti-rabbit IgG antibodies. The volume of distribution was larger for Y-90 than for In-111-labeled antiferritin (160 versus 110% of estimated blood volume). Accidentally extravasated rabbit IgG was rapidly catabolized in perivascular tissues with an effective half-life of less than 35 h. Slower catabolism was noted for rabbit IgG in blood (t(1/2) = 40 h), liver (t(1/2) = 62 h) or tumor (t(1/2) = 40-80 h). Twelve of 13 patients with an effective tumor half-life > 57 h showed a tumor response. I.v. administered polyclonal rabbit antihuman ferritin, labeled with In-111 or Y-90 is stable in vivo and targets Hodgkin's disease. Intravascular Y-90 causes a vascular leak and a larger volume of distribution for antiferritin. Elevated Y-90 blood levels at 1 h and a tumor half-life of >57 h predict for hematological toxicity and tumor response, respectively.


Expression and distribution of vanilloid receptor 1 (TRPV1) in the adult rat brain.
The vanilloid receptor (TRPV1 or VR1) is a molecular integrator of various painful stimuli, including capsaicin, acid, and high temperature. It can also be activated by endogenous ligands, like the cannabinoid 1 receptor (CB1) agonist anandamide. TRPV1 is well characterized at the terminals of sensory nerves involved in the pain pathway. There is also evidence that TRPV1 is expressed in the brain but little is known about its function. Here, using commercially available specific antibodies to investigate the localization of TRPV1 in the brain of the rat, we report that TRPV1 was expressed in hippocampus, cortex, cerebellum, olfactory bulb, mesencephalon and hindbrain. Immunohistochemical analyses showed high expression in the cell bodies and dendrites of neurons in the hippocampus and in the cortex. To address the question of subcellular localization, immunoelectronmicroscopy was used. TRPV1-like staining was detected in the synapses (mostly, but not exclusively in post-synaptic dendritic spines), on the end feet of astrocytes and in pericytes. In summary, TRPV1 expression shows wide distribution in the brain of the rat, being found in astrocytes and pericytes as well as in neurons. Its localization is consistent with multiple functions within the central nervous system, including the regulation of brain vasculature.


Hemorrhagic Transformation in Patients With Acute Ischemic Stroke and Atrial Fibrillation: Time to Initiation of Oral Anticoagulant Therapy and Outcomes.
Background In patients with acute ischemic stroke and atrial fibrillation, early anticoagulation prevents ischemic recurrence but with the risk of hemorrhagic transformation ( HT ). The aims of this study were to evaluate in consecutive patients with acute stroke and atrial fibrillation (1) the incidence of early HT, (2) the time to initiation of anticoagulation in patients with HT , (3) the association of HT with ischemic recurrences, and (4) the association of HT with clinical outcome at 90 days. Methods and Results HT was diagnosed by a second brain computed tomographic scan performed 24 to 72 hours after stroke onset. The incidence of ischemic recurrences as well as mortality or disability (modified Rankin Scale scores >2) were evaluated at 90 days. Ischemic recurrences were the composite of ischemic stroke, transient ischemic attack, or systemic embolism. Among the 2183 patients included in the study, 241 (11.0%) had HT . Patients with and without HT initiated anticoagulant therapy after a mean 23.3 and 11.6 days, respectively, from index stroke. At 90 days, 4.6% (95% confidence interval, 2.3-8.0) of the patients with HT had ischemic recurrences compared with 4.9% (95% confidence interval, 4.0-6.0) of those without HT ; 53.1% of patients with HT were deceased or disabled compared with 35.8% of those without HT . On multivariable analysis, HT was associated with mortality or disability (odds ratio, 1.71; 95% confidence interval, 1.24-2.35). Conclusions In patients with HT , anticoagulation was initiated about 12 days later than patients without HT . This delay was not associated with increased detection of ischemic recurrence. HT was associated with increased mortality or disability.


Odontogenic abscesses in rhesus macaques (Macaca mulatta) of Cayo Santiago.
Odontogenic abscesses are one of the most common dental diseases causing maxillofacial skeletal lesions. They affect the individual's ability to maintain the dental structures necessary to obtain adequate nutrition for survival and reproduction. In this study, the prevalence and pattern of odontogenic abscesses in relation to age, sex, matriline, and living periods were investigated in adult rhesus macaque skeletons of the free-ranging colony on Cayo Santiago, Puerto Rico. The skulls used for this study were from the skeletons of 752 adult rhesus macaques, aged 8-31 years, and born between 1951 and 2000. They came from 66 matrilines ranging from 1 to 88 individuals. Fistulae or skeletal lesions caused by odontogenic abscesses drainage, carious lesions, tooth fractures, tooth loss, and alveolar resorption were evaluated visually. Seventy-two specimens (9.57%) had odontogenic abscesses of varying severity. Males had a significantly higher prevalence than females. The prevalence of odontogenic abscesses in several matrilines was significantly higher than in the population as a whole. Animals born between 1950 and 1965 tended to have a higher prevalence of odontogenic abscesses than those born in later periods. These results suggest that oral pathologies, such as dental and periodontal abscesses in rhesus macaques are fairly common, which may indicate familial effects interwoven with ecological and social factors. The closeness of the rhesus and human genomes allows insights to understand of the epidemiology of these diseases in the human population. Further assessment of the role played by environmental and familial factors on rhesus oral health and disease are warranted.


A revised biosynthetic pathway for phosphatidylinositol in Mycobacteria.
For the last decade, it has been believed that phosphatidylinositol (PI) in mycobacteria is synthesized from free inositol and CDP-diacylglycerol by PI synthase in the presence of ATP. The role of ATP in this process, however, is not understood. Additionally, the PI synthase activity is extremely low compared with the PI synthase activity of yeast. When CDP-diacylglycerol and [(14)C]1L-myo-inositol 1-phosphate were incubated with the cell wall components of Mycobacterium smegmatis, both phosphatidylinositol phosphate (PIP) and PI were formed, as identified by fast atom bombardment-mass spectrometry and thin-layer chromatography. PI was formed from PIP by incubation with the cell wall components. Thus, mycobacterial PI was synthesized from CDP-diacylglycerol and myo-inositol 1-phosphate via PIP, which was dephosphorylated to PI. The gene-encoding PIP synthase from four species of mycobacteria was cloned and expressed in Escherichia coli, and PIP synthase activity was confirmed. A very low, but significant level of free [(3)H]inositol was incorporated into PI in mycobacterial cell wall preparations, but not in recombinant E. coli cell homogenates. This activity could be explained by the presence of two minor PI metabolic pathways: PI/inositol exchange reaction and phosphorylation of inositol by ATP prior to entering the PIP synthase pathway.


JunB Controls Intestinal Effector Programs in Regulatory T Cells.
Foxp3-expressing regulatory T (Treg) cells are critical mediators of immunological tolerance to both self and microbial antigens. Tregs activate context-dependent transcriptional programs to adapt effector function to specific tissues; however, the factors controlling tissue-specific gene expression in Tregs remain unclear. Here, we find that the AP-1 transcription factor JunB regulates the intestinal adaptation of Tregs by controlling select gene expression programs in multiple Treg subsets. Treg-specific ablation of JunB results in immune dysregulation characterized by enhanced colonic T helper cell accumulation and cytokine production. However, in contrast to its classical binding-partner BATF, JunB is dispensable for maintenance of effector Tregs as well as most specialized Treg subsets. In the Peyer's patches, JunB activates a transcriptional program facilitating the maintenance of CD25- Tregs, leading to the complete loss of T follicular regulatory cells in the absence of JunB. This defect is compounded by loss of a separate effector program found in both major colonic Treg subsets that includes the cytolytic effector molecule granzyme B. Therefore, JunB is an essential regulator of intestinal Treg effector function through pleiotropic effects on gene expression.


Poliomyelitis in chimpanzees; studies in homologous and heterologous immunity following inapparent infection.
The response of eighteen chimpanzees to poliomyelitis virus administered orally and cutaneously has been studied. There were no signs of weakness or paralysis in any of the animals. Sixteen of the eighteen (89 per cent) became infected as measured by intestinal carriage of virus. Only one of twelve was reinfected when challenged with an homologous (or homotypic) strain by the same routes, but seven of ten were reinfected on heterologous challenge. A correlation between the development of humoral antibodies and resistance to reinfection was demonstrated. During the course of these experiments, two chimpanzees acquired inapparent poliomyelitis by accidental contagion in the laboratory.


An acute rise in intraluminal pressure shifts the mediator of flow-mediated dilation from nitric oxide to hydrogen peroxide in human arterioles.
Endothelial nitric oxide (NO) is the primary mediator of flow-mediated dilation (FMD) in human adipose microvessels. Impaired NO-mediated vasodilation occurs after acute and chronic hypertension, possibly due to excess generation of reactive oxygen species (ROS). The direct role of pressure elevation in this impairment of human arteriolar dilation is not known. We tested the hypothesis that elevation in pressure is sufficient to impair FMD. Arterioles were isolated from human adipose tissue and cannulated, and vasodilation to graded flow gradients was measured before and after exposure to increased intraluminal pressure (IILP; 150 mmHg, 30 min). The mediator of FMD was determined using pharmacological agents to reduce NO [N(G)-nitro-l-arginine methyl ester (l-NAME), 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO)], or H2O2 [polyethylene glycol (PEG)-catalase], and mitochondrial (mt) ROS was quantified using fluorescence microscopy. Exposure to IILP decreased overall FMD (max %dilation: 82.7 ± 4.9 vs. 62 ± 5.6; P < 0.05). This dilation was abolished by treatment with l-NAME prepressure and PEG-catalase after IILP (max %dilation: l-NAME: 23.8 ± 6.1 vs. 74.8 ± 8.6; PEG-catalase: 71.8 ± 5.9 vs. 24.6 ± 10.6). To examine if this change was mediated by mtROS, FMD responses were measured in the presence of the complex I inhibitor rotenone or the mitochondrial antioxidant mitoTempol. Before IILP, FMD was unaffected by either compound; however, both inhibited dilation after IILP. The fluorescence intensity of mitochondria peroxy yellow 1 (MitoPY1), a mitochondria-specific fluorescent probe for H2O2, increased during flow after IILP (%change from static: 12.3 ± 14.5 vs. 127.9 ± 57.7). These results demonstrate a novel compensatory dilator mechanism in humans that is triggered by IILP, inducing a change in the mediator of FMD from NO to mitochondria-derived H2O2.


Implementation of intermittent theta burst stimulation compared to conventional repetitive transcranial magnetic stimulation in patients with treatment resistant depression: A cost analysis.
Repetitive transcranial magnetic stimulation (rTMS) is an evidence-based treatment for depression that is increasingly implemented in healthcare systems across the world. A new form of rTMS called intermittent theta burst stimulation (iTBS) can be delivered in 3 min and has demonstrated comparable effectiveness to the conventional 37.5 min 10Hz rTMS protocol in patients with depression. To compare the direct treatment costs per course and per remission for iTBS compared to 10Hz rTMS treatment in depression. We conducted a cost analysis from a healthcare system perspective using patient-level data from a large randomized non-inferiority trial (THREE-D). Depressed adults 18 to 65 received either 10Hz rTMS or iTBS treatment. Treatment costs were calculated using direct healthcare costs associated with equipment, coils, physician assessments and technician time over the course of treatment. Cost per remission was estimated using the proportion of patients achieving remission following treatment. Deterministic sensitivity analyses and non-parametric bootstrapping was used to estimate uncertainty. From a healthcare system perspective, the average cost per patient was USD$1,108 (SD 166) for a course of iTBS and $1,844 (SD 304) for 10Hz rTMS, with an incremental net savings of $735 (95% CI 688 to 783). The average cost per remission was $3,695 (SD 552) for iTBS and $6,146 (SD 1,015) for 10Hz rTMS, with an average incremental net savings of $2,451 (95% CI 2,293 to 2,610). The shorter session durations and treatment capacity increase associated with 3 min iTBS translate into significant cost-savings per patient and per remission when compared to 10Hz rTMS.


Cardiac function during rest and exercise in patients with mitral valve prolapse. Role of radionuclear angiocardiography.
The clinical features of mitral valve prolapse syndrome and ischemic coronary disease overlap, making differentiation of the two conditions difficult. Furthermore, many patients have both conditions. This study assessed changes in ventricular function during rest and exercise in patients with mitral valve prolapse alone and in patients with prolapse and concomitant coronary artery disease. Twelve patients with angiographically documented mitral valve prolapse and normal coronary anatomy and 11 patients with normal coronary anatomy and no mitral valve prolapse had increased ejection fraction and demonstrated no wall motion abnormality during exercise. Changes in ventricular function during exercise in 11 additional patients with mitral valve prolapse demonstrated on echocardiography were similar to those in the group with mitral valve prolapse and normal coronary anatomy seen on angiography. In contrast, 6 of 11 patients with mitral valve prolapse and coronary arterial stenosis demonstrated on angiography had a decreased ejection fraction and exhibited wall motion abnormalities during exercise. These results suggest that mitral valve prolapse alone has no detrimental effect on ventricular function during rest and exercise and that exercise-induced abnormalities in ventricular function are related to the presence and severity of coronary artery disease and not to mitral valve prolapse.


A new function for the LDL receptor: transcytosis of LDL across the blood-brain barrier.
Lipoprotein transport across the blood-brain barrier (BBB) is of critical importance for the delivery of essential lipids to the brain cells. The occurrence of a low density lipoprotein (LDL) receptor on the BBB has recently been demonstrated. To examine further the function of this receptor, we have shown using an in vitro model of the BBB, that in contrast to acetylated LDL, which does not cross the BBB, LDL is specifically transcytosed across the monolayer. The C7 monoclonal antibody, known to interact with the LDL receptor-binding domain, totally blocked the transcytosis of LDL, suggesting that the transcytosis is mediated by the receptor. Furthermore, we have shown that cholesterol-depleted astrocytes upregulate the expression of the LDL receptor at the BBB. Under these conditions, we observed that the LDL transcytosis parallels the increase in the LDL receptor, indicating once more that the LDL is transcytosed by a receptor-mediated mechanism. The nondegradation of the LDL during the transcytosis indicates that the transcytotic pathway in brain capillary endothelial cells is different from the LDL receptor classical pathway. The switch between a recycling receptor to a transcytotic receptor cannot be explained by a modification of the internalization signals of the cytoplasmic domain of the receptor, since we have shown that LDL receptor messengers in growing brain capillary ECs (recycling LDL receptor) or differentiated cells (transcytotic receptor) are 100% identical, but we cannot exclude posttranslational modifications of the cytoplasmic domain, as demonstrated for the polymeric immunoglobulin receptor. Preliminary studies suggest that caveolae are likely to be involved in the potential transport of LDL from the blood to the brain.


Intraindividual Temporal miRNA Variability in Serum, Plasma, and White Blood Cell Subpopulations.
Blood microRNAs (miRNAs) are ideal biomarkers, and blood derivatives are often collected in the scope of miRNA research projects. However, knowledge of temporal variations of miRNAs in healthy individuals is lacking. In this study, miRNA variability was measured over a 1-year period in different blood derivatives, collected every 2-3 months from two healthy donors. There is a continuum of intraindividual temporal variability, with particularly stable (coefficient of variation [CV] <20%-30%) and particularly unstable (CV >100%-130%) miRNAs in serum, plasma, and specific white blood cell subpopulations. The temporal intraindividual variability of miRNAs should be taken into consideration in experimental design of biospecimen collections and validation of diagnostic biomarkers.


Differentiating Schistosoma haematobium from Schistosoma magrebowiei and other closely related schistosomes by polymerase chain reaction amplification of a species specific mitochondrial gene.
Schistosoma haematobium infection afflicts about 150 million people in 53 countries in Africa and the Middle East. In many endemic areas, S. haematobium is sympatric with Schistosoma bovis, Schistosoma mattheei, Schistosoma curassoni, Schistosoma intercalatum and Schistosoma magrebowiei, its closely related species. In addition, they also develop in the same intermediate snail hosts. Since these schistosome species often infect snails inhabiting the same bodies of water, examining cercariae or infected snails for estimating transmission of S. haematobium is always confounded by the need to differentially identify S. haematobium from these other species. Recently, differentiating S. haematobium by polymerase chain reaction (PCR) from S. bovis, S. mattheei, S. curassoni and S. intercalatum, but not from S. magrebowiei was reported. However, to be able to evaluate residual S. haematobium transmission after control interventions in areas where S. haematobium may be sympatric with S. magrebowiei, a differential tool for accurate monitoring of infected snails is needed. Thus in this study, we developed a new PCR assay using a pair of primers, ShND-1/ShND-2, to amplify a target sequence of 1117 bp (GenBank accession number KF834975) from S. haematobium mitochondrion complete genome (GenBank accession number DQ157222). Sensitivity of the assay was determined by PCR amplification of different concentrations of S. haematobium gDNA serially diluted from 10ng to 0.1pg. For assay specificity, different concentrations of gDNA from S. haematobium and the other schistosome species, 20 positive urine samples and five controls as well as 20 infected snails were subjected to PCR amplification, while some of the PCR products were sequenced. The assay detected up to 1pg of S. haematobium gDNA, while a differential identification of S. haematobium DNA content from other closely related species was achieved when applied to urine and naturally infected snails. When a protein-protein blast search was carried out using Blastp, the amplified sequence was found to encode a protein that shows a 100% similarity with S. haematobium nicotinamide adenine dinucleotide dehydrogenase subunit 3 (GenBank accession number YP_626524.1). The PCR assay was sensitive, specific and was able to successfully differentiate S. haematobium from S. magrebowiei, in addition to its other closely related animal infective schistosome species.


Multiple drilling compared with standard core decompression for avascular necrosis of the femoral head in sickle cell disease patients.
Sickle cell disease (SCD) is the most common cause of avascular necrosis of femoral head (ANFH) in childhood. Advances in medical treatment led to improved life expectancy of such patients. SCD-related ANFH frequently progress to total collapse of the femoral head necessitating hip replacement. However, SCD patient are at more risk of intra- and post-operative complications and suboptimal outcome of total hip arthroplasty. Hence, it is imperative to preserve the femoral head as long as possible. Between September 1992 and June 2007, 94 core decompression procedures were done to SCD patients who had modified Ficat stage I, IIA and IIB ANFH. Sixty one patients underwent a classical 8-mm drilling and 33 patients underwent 3.2-mm diameter MD technique. Patients were followed up for minimum of 2 years and were evaluated for clinical and Harris Hip Score improvement and for radiological progression. All 19 hips that had Ficat stage I had significant reduction of pain and improvement of Harris Hip Score. No patient has required further surgery. Among the 39 hips with Ficat IIA at time of procedure, 80 % of hips which underwent CD and 78 % of MD cases had significant reduction of pain and improvement of HHS. Those patients showed no radiographic progression of the disease. The remaining, 20 % CD and 22 % MD eventually progressed radiologically to grade III or grade IV and had HHS less than 75 at last visit. In the 36 cases with Ficat IIB, 52 % CD and 52.8 % MC had significant reduction of pain and improvement of HHS. The rest showed no improvement in pain and function, and progressed to stage IV; 11 of them underwent THA and one patient refused surgery. While multiple drilling is safer and less invasive than single coring in SCD, there is no statistically significant difference in outcome or complication rate between both procedures done for ANFH in patients with SCD.


Future Dietitian 2025: informing the development of a workforce strategy for dietetics.
Healthcare is changing and the professions that deliver it need to adapt and change too. The aim of this research was to inform the development of a workforce strategy for Dietetics for 2020-2030. This included an understanding of the drivers for change, the views of stakeholders and recommendations to prepare the profession for the future. The research included three phases: (i) establishing the context which included a literature and document review (environmental scan); (ii) discovering the profession and professional issues using crowd-sourcing technology; and (iii) articulating the vision for the future using appreciative inquiry. The environmental scan described the current status of the dietetic profession, the changing healthcare environment, the context in which dietitians work and what future opportunities exist for the profession. The online conversation facilitated by crowd-sourcing technology asked the question: 'How can dietitians strengthen their future role, influence and impact?' Dietitians and interested stakeholders (726 and 109, respectively) made 6130 contributions. Seven priorities were identified and fed into the appreciative inquiry event. The event bought together 54 dietitians and analysis of the discussions generated five themes: (i) professional identity; (ii) strong foundations-creating structure and direction for the profession; (iii) amplifying visibility and influence; (iv) embracing advances in science and technology; and (v) career advancement and emerging opportunities. A series of recommendations were made for the next steps in moving the workforce to a new future. The future for dietetics looks bright, embracing technology, as well as exploring different ways of working and new opportunities, as this dynamic profession continues to evolve.


How are sexual health clinics in England managing men who have sex with men who refuse to be tested for HIV?
The aim of this study was to ascertain the existence of written policies and common clinical practices in sexual health clinics in England for the management of men who have sex with men who refuse to test for HIV. All sexual health clinics in England ( n = 223) were invited to complete an online questionnaire in August-September 2014. The questionnaire covered the four domains of clinic policies, management practices, training and monitoring. We assess differences by region. Overall, 92 clinics participated. Only three clinics reported having a written policy. In contrast, most reported having a common agreed practice (94% in London vs. 71.6% elsewhere). When encountering a refusal, 72.2% of the London clinics and 53.4% of the clinics from elsewhere offered a less invasive option. Few clinics (17.4%) provided information on home sampling kits and 74.4% informed about other testing options. Eighty-seven per cent of the clinics recorded the occurrence of refusals, but only 37.8% reviewed the collected data. Providing staff with training was more common in London (94.1% vs. 73.8%). Clear policies should be developed to guide professionals when encountering men who have sex with men who refuse an HIV test. Offering less invasive testing options and information on alternative testing options could be easily introduced into routine practice. Efforts should be made to review monitoring data in order to identify implications of test refusals and introduce improvements in management of refusals.


An improved environmental DNA assay for bull trout (Salvelinus confluentus) based on the ribosomal internal transcribed spacer I.
The majority of environmental DNA (eDNA) assays for vertebrate species are based on commonly analyzed regions of the mitochondrial genome. However, the high degree of mitochondrial similarity between two species of charr (Salvelinus spp.), southern Dolly Varden (S. malma lordii) and bull trout (Salvelinus confluentus), precludes the development of a mitochondrial eDNA assay to distinguish them. Presented here is an eDNA assay to detect bull trout based on the first ribosomal internal transcribed spacer (ITSI), a nuclear marker. This assay successfully detects bull trout and avoids detecting Dolly Varden as well as brook trout (S. fontinalis), Arctic char (S. alpinus), and lake trout (S. namaycush). In addition, this assay was compared with an extensively used mitochondrial bull trout assay and it was found that the ITSI-based assay produced higher detectability. Our results suggest this assay should out-perform the published mtDNA assay across the range of bull trout, while the added specificity allows reliable bull trout detection in areas where bull trout co-occur with other charr such as Dolly Varden. While clearly a superior assay in this instance, basing assays on ITSI is not without problems. For vertebrates, there are fewer ITSI sequences available than commonly sequenced regions of the mitochondrial genome. Thus, the initial in silico screening of candidate assays must be preceded by much more extensive sampling and sequencing of sympatric or closely related taxa. Further, all copies of the internal transcribed spacers within an individual may not be identical, which can lead to complications. Lastly, the copy number for ITSI varies widely across taxa; the greater detectability associated with this assay cannot be assumed for other species.


Multigeneration toxicity of Geunsami® (a glyphosate-based herbicide) to Allonychiurus kimi (Lee) (Collembola) from sub-individual to population levels.
Glyphosate-based herbicide (GBH) is the most widely used herbicide worldwide and has long been considered to have significantly low toxicity to non-target soil invertebrates based on short-term toxicity tests (<56 d). However, long-term GBH toxicity assessment is necessary as GBH is repeatedly applied in the same field annually because of the advent of glyphosate-resistant crops. In this study, a multigeneration test was conducted where Allonychiurus kimi (Collembola) was exposed to GBH for three generations (referred to as F0, F1, and F2) to evaluate the long-term toxic effect. The endpoints used were adult survival and juvenile production for the individual level toxicity assessment. Phospholipid profile and population age structure were the endpoints used for sub-individual and population levels, respectively. GBH was observed to have no negative effects on adult survivals of all generations, but juvenile production was found to decrease in a concentration-dependent manner, with EC50s being estimated as 572.5, 274.8, and 59.8 mg a.i. kg-1 in the F0, F1, and F2 generations, respectively. The age structure of A. kimi population produced in the test of all generations was altered by GBH exposure, mainly because of the decrease in the number of young juveniles. Further, differences between the phospholipid profiles of the control and GBH treatments became apparent over generations, with PA 16:0, PA 12:0, and PS 42:0 lipids not being detected at the highest concentration of 741 mg kg-1 in F2. Considering all our findings from sub-individual to population levels, repeated and long-term use of GBH could have significantly higher negative impacts on non-target soil organisms than expected.


Large-scale prediction of Saccharomyces cerevisiae gene function using overlapping transcriptional clusters.
Genome sequencing has led to the discovery of tens of thousands of potential new genes. Six years after the sequencing of the well-studied yeast Saccharomyces cerevisiae and the discovery that its genome encodes approximately 6,000 predicted proteins, more than 2,000 have not yet been characterized experimentally, and determining their functions seems far from a trivial task. One crucial constraint is the generation of useful hypotheses about protein function. Using a new approach to interpret microarray data, we assign likely cellular functions with confidence values to these new yeast proteins. We perform extensive genome-wide validations of our predictions and offer visualization methods for exploration of the large numbers of functional predictions. We identify potential new members of many existing functional categories including 285 candidate proteins involved in transcription, processing and transport of non-coding RNA molecules. We present experimental validation confirming the involvement of several of these proteins in ribosomal RNA processing. Our methodology can be applied to a variety of genomics data types and organisms.


In vivo kinematics of the knee after anterior cruciate ligament reconstruction: a clinical and functional evaluation.
Recent follow-up studies have reported a high incidence of joint degeneration in patients with anterior cruciate ligament reconstruction. Abnormal kinematics after anterior cruciate ligament reconstruction have been thought to contribute to the degeneration. Anterior cruciate ligament reconstruction, which was designed to restore anterior knee laxity under anterior tibial loads, does not reproduce knee kinematics under in vivo physiological loading conditions. Controlled laboratory study. Both knees of 7 patients with complete unilateral rupture of the anterior cruciate ligament were magnetic resonance imaged, and 3D models were constructed from these images. The anterior cruciate ligament of the injured knee was arthroscopically reconstructed using a bone-patellar tendon-bone autograft. Three months after surgery, the kinematics of the intact contralateral and reconstructed knees were measured using a dual-orthogonal fluoroscopic system while the subjects performed a single-legged weightbearing lunge. The anterior laxity of both knees was measured using a KT-1000 arthrometer. The anterior laxity of the reconstructed knee as measured with the arthrometer was similar to that of the intact contralateral knee. However, under weightbearing conditions, there was a statistically significant increase in anterior translation of the reconstructed knee compared with the intact knee at full extension (approximately 2.9 mm) and 15 degrees (approximately 2.2 mm) of flexion. In addition, there was a mean increase in external tibial rotation of the anterior cruciate ligament-reconstructed knee beyond 30 degrees of flexion (approximately 2 degrees at 30 degrees of flexion), although no statistical significance was detected. The data demonstrate that although anterior laxity was restored during KT-1000 arthrometer testing, anterior cruciate ligament reconstruction did not restore normal knee kinematics under weightbearing loading conditions. Future reconstruction techniques should aim to restore function of the knee under physiological loading conditions.


Oral transmucosal fentanyl citrate for the treatment of breakthrough pain in cancer patients: an overview of its pharmacological and clinical characteristics.
Breakthrough pain is a transitory flare of pain occurring in most cancer patients against a background of otherwise controlled persistent pain. Treatment of breakthrough pain is a challenging phenomenon. Oral transmucosal fentanyl citrate (OTFC; brand name Actiq, Chephalon Inc., West Chester, PA), a new opioid formulation with a unique delivery system, reflects the characteristics of breakthrough pain (rapid onset of action and short duration), making it an effective treatment for cancer patients who already receive opioids and experience flares of pain. This review article aims to present the role of oral transmucosal fentanyl citrate in the management of breakthrough pain in cancer patients. In particular, it is going to discuss the synthesis, clinical pharmacology, pharmacokinetic and pharmacodynamic properties, toxicity, and clinical efficacy of this novel agent.


Two Subgroups of Porcine Circovirus 2 Appearing among Pigs in Southern China.
We report here two complete genome sequences of porcine circovirus 2. The complete genomes of BH5 and BH6 were amplified and analyzed. Sequence analysis demonstrated that strains BH5 and BH6 belonged to PCV2d and PCV2b, respectively. Knowledge regarding the complete genome sequences of strains BH5 and BH6 will be useful for epidemiological surveillance.


Electronic health systems: challenges faced by hospital-based providers.
The purpose of this article is to discuss specific challenges faced by hospitals adopting the use of electronic medical records and implementing electronic health record (EHR) systems. Challenges include user and information technology support; ease of technical use and software interface capabilities; compliance; and financial, legal, workforce training, and development issues. Electronic health records are essential to preventing medical errors, increasing consumer trust and use of the health system, and improving quality and overall efficiency. Government efforts are focused on ways to accelerate the adoption and use of EHRs as a means of facilitating data sharing, protecting health information privacy and security, quickly identifying emerging public health threats, and reducing medical errors and health care costs and increasing quality of care. This article will discuss physician and nonphysician staff training before, during, and after implementation; the effective use of EHR systems' technical features; the selection of a capable and secure EHR system; and the development of collaborative system implementation. Strategies that are necessary to help health care providers achieve successful implementation of EHR systems will be addressed.


Impact of academic affiliation and training on knowledge of hereditary colorectal cancer.
Knowledge about hereditary colorectal cancer (CRC) can aid cancer screening and prevention in high-risk patients. Genetic testing, once conducted primarily at academic centers, is now routinely performed in a variety of clinics. Nonacademic physicians may not be aware of hereditary CRC standards of care. From August to November 2012, a survey was administered to predominantly primary care physicians evaluating academic center affiliation, past training in genetics and knowledge regarding hereditary CRC. One hundred forty physicians completed the survey. Knowledge of hereditary CRC was neither associated with academic affiliation nor with training during medical school or residency, but with continuing medical education (CME) training. Those with CME training were more likely to know that screening could be enhanced for patients with a hereditary cancer risk (OR = 4.49, 95% CI = 1.40-14.38) and that an individual with hereditary CRC would have different screening recommendations (OR = 7.49, 95% CI = 1.37-40.81). Residency training and CME training were associated with more frequent hereditary risk assessment. Genetics training may be associated with physicians' knowledge and assessment of hereditary CRC. Training at the CME level in particular may be integral to the delivery of genetic services in clinical practice.


Early diagnosis of opportunistic systemic fungal and nocardial infections.
An approach to the expediting of the diagnosis of opportunistic systemic mycoses is presented. Communication between clinician and microbiologist is basic to this approach. The importance of the clinical assessment of the individual patient, coupled with a high index of suspicion, is stressed. Our experience with 11 of 42 cases of systemic mycosis over a 28-month period is analysed. For the diagnosis of fungaemia a method for the microscopical examination of peripheral blood is briefly evaluated, and a membrane filter blood culture technique is shown to be valuable, yielding results in 16-24 hours. In the absence of fungaemia the considered microscopical examination of suitable specimens, when feasible, is the most rapid method available. Serological methods may be helpful in early diagnosis, but this is often hampered by the absence of baseline sera and by the lengthy nature of some tests. Newer indirect methods such as gas chromatography are being developed but have not yet been used routinely.


In vitro production of large single-stranded templates for DNA sequencing.
A method has been developed for the preparation of large single-stranded DNA sequencing templates from primary cloning plasmids or cosmids. The method utilizes the separate action of T7 Gene 6 exonuclease and exonuclease III to generate large quantities of single-stranded template for each strand of a large-cloned fragment. Since the procedure is intended for use on primary clones, it avoids the time-consuming subcloning steps associated with most sequencing programs. The procedure also has the advantage of avoiding clone instability problems associated with subcloning in M13.


Use of three-dimensional microscopy with multiple oblique illumination in evaluating cytopathology preparations.
To analyze whether the use of multiple oblique illumination (MOI), in contrast to axial illumination (AI), improves imagery in bright field microscopy. Specimens containing thick cell clusters, such as cervical Pap smears, are often misread because of out-of-focus cell clusters. We hypothesized that visualization of these specimens with MOI would enhance this information as compared with AI. Micrometer images and Pap smears were analyzed in focus, and 8 and 40 microns out of focus by MOI and AI. All fields were captured to a remote computer, histograms were constructed, and mean light intensity was calculated. Mathematical formulation was used to define the histogram distribution of the micrometer images. Statistical analysis was performed using one-way ANOVA and Newman-Keuls test. K(focus) was improved (P < .01) and at 20 and 40 microns out of focus, mean light intensities were greater (P < .003, P < .05, respectively) with MOI as compared to AI. MOI improves out-of-focus information by increasing light penetration through the specimen and enhancing contrast and resolution.


In vivo brain electrophoresis - a novel method for chemotherapy of CNS diseases.
The blood-brain barrier (BBB) is a protective mechanism that does its job superbly. So much so, that hitherto, brain chemotherapy has been limited by it. In fact, very few agents are effective against brain disease due to the inherent difficulties of penetrating the BBB. We describe a novel, extremely focused method for delivering drugs to specific diseased areas. This innovative method directly delivers putative substances to the pathological area, bypassing the BBB. Treatment of brain diseases could be improved by targeted, controlled delivery of therapeutic substances to diseased cerebral areas. Our described novel method - in vivo electrophoresis - achieves this. This technique was evaluated in beagles after craniotomy was performed and a custom-designed plate with electrodes inserted. The delivery of charged substances to selected areas with predictably guided movement was achieved via a created electrical field. Gadolinium, a compound unable to cross the BBB, was injected intracerebrally whereas an electrical field was created using the implanted electrodes surrounding the injection area. The electrical field-guided Gadolinium movement was evaluated using MRI. Gadolinium was moved predictably using the created electrical field without complications. The experiment successfully demonstrated controlled movement of the substance. This technique can significantly change treatment of brain diseases because substances: i) may be moved in a controlled, predictable way - exponentially increasing therapeutic interactions with the target; and ii) no longer need to conform to constraints dictated by the BBB (molecular mass < 500 d; lipophilic), thereby increasing potential number of usable substances.


Electrophysiologic evidence of increased distractibility after dorsolateral prefrontal lesions.
Patients with left prefrontal lesions and control subjects showed enhanced event-related potentials (ERPs) to attended tone sequences presented in a dichotic attention task. ERP enhancements were comparable at short and long interstimulus intervals (ISIs), and did not depend upon whether attended stimuli were preceded by other attended stimuli or by distracting stimuli in the opposite ear. In contrast, patients with right prefrontal lesions showed absent ERP attention effects to contralateral (left ear) tones at all ISIs, and reduced attention effects to ipsilateral tones at long ISIs and when these were preceded by distracting sounds. The results are consistent with an asymmetric organization of dorsolateral prefrontal cortex, and indicate that increased distractibility may contribute to the attention disorders that follow prefrontal lesions.


Preparing raw clinical data for publication: guidance for journal editors, authors, and peer reviewers.
In recognition of the benefits of transparent reporting, many peer-reviewed journals require that their authors be prepared to share their raw, unprocessed data with other scientists and/or state the availability of raw data in published articles. But little information on how data should be prepared for publication - or sharing - has emerged. In clinical research patient privacy and consent for use of personal health information are key considerations, but agreed-upon definitions of what constitutes anonymised patient information do not appear to have been established. We aim to address this issue by providing practical guidance for those involved in the publication process, by proposing a minimum standard for de-identifying datasets for the purposes of publication in a peer-reviewed biomedical journal, or sharing with other researchers. Basic advice on file preparation is provided along with procedural guidance on prospective and retrospective publication of raw data, with an emphasis on randomised controlled trials.


Bacterial adhesion and colonization differences between zirconium oxide and titanium alloys: an in vivo human study.
The purpose of this study was to compare zirconium oxide and titanium alloys with respect to their tendency to adhesion and colonization of two periodontal pathogens on both hard surfaces and on soft tissues in vivo. The present study was designed as a prospective stratified randomized controlled clinical trial. Patients were scheduled to receive two implants with different types of abutments in the posterior mandible. Three months after implant placement, titanium and zirconium abutments were connected. Five weeks after abutment connections, the abutments were removed, probing depth measurements were recorded, and gingival biopsy samples were obtained. Abutments and biopsy specimens were analyzed by reverse-transcriptase polymerase chain reaction to compare the DNA copy numbers of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and total bacteria. The surface free energy of the abutments was calculated by sesile water drop before replacement. No statistically significant differences were found between probing depths or DNA copy numbers of A actinomycetemcomitans, P gingivalis, and total bacteria both for both titanium alloys and zirconium oxide surfaces and the biops specimens obtained from their buccal gingival. With respect to the surface free energy of zirconium and titanium abutments, zirconium abutments showed lower surface free energy than titanium abutments. The results of this study showed that zirconium oxide surfaces have comparable properties to titanium alloy surfaces in their tendency to adhesion and colonization of two periodontal pathogens on both hard surfaces and in soft tissues.


Antifungal activity and fungal metabolism of steroidal glycosides of Easter lily (Lilium longiflorum Thunb.) by the plant pathogenic fungus, Botrytis cinerea.
Botrytis cinerea Pers. Fr. is a plant pathogenic fungus and the causal organism of blossom blight of Easter lily (Lilium longiflorum Thunb.). Easter lily is a rich source of steroidal glycosides, compounds which may play a role in the plant-pathogen interaction of Easter lily. Five steroidal glycosides, including two steroidal glycoalkaloids and three furostanol saponins, were isolated from L. longiflorum and evaluated for fungal growth inhibition activity against B. cinerea, using an in vitro plate assay. All of the compounds showed fungal growth inhibition activity; however, the natural acetylation of C-6''' of the terminal glucose in the steroidal glycoalkaloid, (22R,25R)-spirosol-5-en-3β-yl O-α-L-rhamnopyranosyl-(1→2)-[6-O-acetyl-β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside (2), increased antifungal activity by inhibiting the rate of metabolism of the compound by B. cinerea. Acetylation of the glycoalkaloid may be a plant defense response to the evolution of detoxifying mechanisms by the pathogen. The biotransformation of the steroidal glycoalkaloids by B. cinerea led to the isolation and characterization of several fungal metabolites. The fungal metabolites that were generated in the model system were also identified in Easter lily tissues infected with the fungus by LC-MS. In addition, a steroidal glycoalkaloid, (22R,25R)-spirosol-5-en-3β-yl O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside (6), was identified as both a fungal metabolite of the steroidal glycoalkaloids and as a natural product in L. longiflorum for the first time.


Alpha-glycosylceramides enhance the antitumor cytotoxicity of hepatic lymphocytes obtained from cancer patients by activating CD3-CD56+ NK cells in vitro.
Alpha-glycosylceramides, such as alpha-galactosylceramide and alpha-glucosylceramide, induce antitumor immunity in various murine cancer models. In the murine hepatic metastasis model, V alpha 14 TCR+NK1.1+ T cells, which accumulate preferentially in the liver, are considered to play a key role in the induction of antitumor immunity by alpha-glycosylceramides. We recently reported that V alpha 24 TCR+ NKT cells, the human homologues of murine V alpha 14 TCR+NK1.1+ cells, are rarely seen among freshly isolated human hepatic lymphocytes. Therefore, it is important to examine whether alpha-glycosylceramides also enhance the antitumor cytotoxicity of human hepatic lymphocytes, as they have been shown to do in murine systems, to determine the usefulness of alpha-glycosylceramides in cancer immunotherapy in humans. Here, we show that alpha-glycosylceramides greatly enhance the cytotoxicity of human hepatic lymphocytes obtained from cancer patients against the tumor cell lines, K562 and Colo201, in vitro. The direct effector cells of the elicited cytotoxicity were CD3-CD56+ NK cells. Even though V alpha 24 TCR+NKT cells proliferated remarkably in response to alpha-glycosylceramides, they did not contribute directly to the cytotoxicity. Our observations strongly suggest the potential usefulness of alpha-glycosylceramides for immunotherapy of liver cancer in humans based on their ability to activate CD3-CD56+ NK cells in the liver.


Human-interpretable image features derived from densely mapped cancer pathology slides predict diverse molecular phenotypes.
Computational methods have made substantial progress in improving the accuracy and throughput of pathology workflows for diagnostic, prognostic, and genomic prediction. Still, lack of interpretability remains a significant barrier to clinical integration. We present an approach for predicting clinically-relevant molecular phenotypes from whole-slide histopathology images using human-interpretable image features (HIFs). Our method leverages >1.6 million annotations from board-certified pathologists across >5700 samples to train deep learning models for cell and tissue classification that can exhaustively map whole-slide images at two and four micron-resolution. Cell- and tissue-type model outputs are combined into 607 HIFs that quantify specific and biologically-relevant characteristics across five cancer types. We demonstrate that these HIFs correlate with well-known markers of the tumor microenvironment and can predict diverse molecular signatures (AUROC 0.601-0.864), including expression of four immune checkpoint proteins and homologous recombination deficiency, with performance comparable to 'black-box' methods. Our HIF-based approach provides a comprehensive, quantitative, and interpretable window into the composition and spatial architecture of the tumor microenvironment.


Increased and controlled expression of the Rickettsia prowazekii ATP/ADP translocase and analysis of cysteine-less mutant translocase.
Detailed molecular analysis of the Rickettsia prowazekii ATP/ADP translocase, an obligate exchange transport system that is specific for ATP and ADP, has been extremely difficult due to limited quantities of material available from these obligate intracytoplasmic bacteria and by the toxicity and poor expression in recombinant Escherichia coli expression systems. In this study, a stable and controllable system for the increased expression of the rickettsial ATP/ADP translocase was developed in E. coli where the expression of translocase from the bacteriophage T7 promoter in the pET11a vector led to a 26-fold increase in ATP transport activity and a 34-fold increase in translocase protein as compared to the expression with the native rickettsial promoter in E. coli. When compared to R. prowazekii, ATP transport activity was increased sixfold and membrane translocase was increased threefold. Approximately 24% of the translocase protein produced was localized in an inclusion body fraction. This expression system was then used to determine whether the two cysteine residues in the ATP/ADP translocase were essential for activity or expression. The translocase was modified by oligonucleotide-directed site-specific mutagenesis such that the two cysteines were converted to alanines. The ATP transport properties and ATP/ADP translocase production kinetics, translocase protein concentration and subcellular localization were indistinguishable in the wild-type and mutant strains, proving that cysteines play no functional role in the R. prowazekii ATP/ADP translocase and providing a system suitable for cysteine-scanning mutagenesis.


Leadership in the perspective of Family Health Strategy nurses.
To understand the perception of leadership in the work process and to promote its discussion within the Family Health Strategy scope. A qualitative approach research, guided by the action-research referential, conducted with 15 Familiy Health Strategy nurses in the inland of the state of São Paulo. Data was collected in 2015 in two interdependent phases, interviews and intervention-action, and processed according to the methodological framework of content analysis and the theoretical framework on nursing management. The nurses showed their conceptions of leadership and the inherent challenges of practice: leadership training, overlapping care and management, and management charges. In the discussion of common problems and peer learning, a conception of transformational leadership was constructed. The common and behavioral styles of leadership influenced on the conception of leadership and mobilized the attitudes of the nurses in the work process; however, there is a need for more engagement in the training/qualification of the professionals and the services about leadership.


Seipin localizes at endoplasmic-reticulum-mitochondria contact sites to control mitochondrial calcium import and metabolism in adipocytes.
Deficiency of the endoplasmic reticulum (ER) protein seipin results in generalized lipodystrophy by incompletely understood mechanisms. Here, we report mitochondrial abnormalities in seipin-deficient patient cells. A subset of seipin is enriched at ER-mitochondria contact sites (MAMs) in human and mouse cells and localizes in the vicinity of calcium regulators SERCA2, IP3R, and VDAC. Seipin association with MAM calcium regulators is stimulated by fasting-like stimuli, while seipin association with lipid droplets is promoted by lipid loading. Acute seipin removal does not alter ER calcium stores but leads to defective mitochondrial calcium import accompanied by a widespread reduction in Krebs cycle metabolites and ATP levels. In mice, inducible seipin deletion leads to mitochondrial dysfunctions preceding the development of metabolic complications. Together, these data suggest that seipin controls mitochondrial energy metabolism by regulating mitochondrial calcium influx at MAMs. In seipin-deficient adipose tissue, reduced ATP production compromises adipocyte properties, contributing to lipodystrophy pathogenesis.


Quantitative assessment of age-related macular degeneration using parametric modeling of the leakage transfer function: preliminary results.
Age-related macular degeneration (AMD) is a major cause of blindness and visual impairment in older adults. The wet form of the disease is characterized by abnormal blood vessels forming a choroidal neovascular membrane (CNV), that result in destruction of normal architecture of the retina. Current evaluation and follow up of wet AMD include subjective evaluation of Fluorescein Angiograms (FA) to determine the activity of the lesion and monitor the progression or regression of the disease. However, this subjective evaluation prevents accurate monitoring of the disease progression or regression in response to a pharmacologic agent. In this work, we present a method that allows objective assessment of the activity of a CNV lesion which can be statistically compared across different patient and time points. The method is based on a hypothesis that the discrepancy in the time-intensity signals among the diseased and normal retinal areas are due to an implicit transfer function whose parameters can be used to characterize the retina. The method begins with parametric modeling of the temporal variation of the lesion and background intensities. Then, the values of the model parameters are used to evaluate the change in the activity of the disease. Preliminary results on five datasets show that the calculated parameters are highly correlated with the Visual Acuity (VA) of the patients.


The acutely infected diabetic foot is not adequately evaluated in an inpatient setting.
To evaluate the standard of evaluation and treatment of the infected diabetic foot ulceration at a 551-bed university teaching institution. A retrospective review of a 4-year consecutive sample (1991-1994). Two hundred fifty-five patients who were admitted to a hospital for care of an infected diabetic foot ulceration. Patients were subdivided into the following 4 dichotomous variables: (1) whether the patient underwent a lower-extremity amputation, (2) whether the patient was admitted once or multiple times, (3) whether the patient underwent intraoperative debridement, and (4) whether the patient was admitted to medical or surgical services. All groups that were evaluated had undergone a less than adequate foot examination. Of the admitted patients, 31.4% did not have their pedal pulses documented; 59.7% of the admitted patients were not evaluated for the presence or absence of protective sensation. Nearly 90% of the wounds were not evaluated for involvement of underlying structures, and foot radiographs were not performed in 32.9% of the patients. There were more blood cultures ordered (62.0%) than wound cultures (51.4%). The results of this study highlight the need for a systematic, detailed lower-extremity examination for every diabetic patient who is admitted to a hospital, particularly those who are admitted with a primary diagnosis that involves a foot complication.


A diagnostic codes-based algorithm improves accuracy for identification of childhood asthma in archival data sets.
While a single but truncated ICD code (493) had been widely used for identifying asthma in asthma care and research, it significantly under-identifies asthma. We aimed to develop and validate a diagnostic codes-based algorithm for identifying asthmatics using Predetermined Asthma Criteria (PAC) as the reference. This is a retrospective cross-sectional study which utilized two different coding systems, the Hospital Adaptation of the International Classification of Diseases, Eighth Revision (H-ICDA) and the International Classification of Diseases, Ninth Revision (ICD-9). The algorithm was developed using two population-based asthma study cohorts, and validated in a validation cohort, a random sample of the 1976-2007 Olmsted County Birth Cohort. Performance of the diagnostic codes-based algorithm for ascertaining asthma status against manual chart review for PAC (gold standard) was assessed by determining both criterion and construct validity. Among eligible 267 subjects of the validation cohort, 50% were male, 70% white, and the median age at last follow-up was 17 (interquartile range, 8.7-24.4) years. Asthma prevalence by PAC through manual chart review was 34%. Sensitivity and specificity of the codes-based algorithm for identifying asthma were 82% and 98% respectively. Associations of asthma-related risk factors with asthma status ascertained by the code-based algorithm were similar to those by the manual review. The diagnostic codes-based algorithm for identifying asthmatics improves accuracy of identification of asthma and can be a useful tool for large scale studies in a setting without automated chart review capabilities.


Crystal structure of the kinesin motor domain reveals a structural similarity to myosin.
Kinesin is the founding member of a superfamily of microtubule based motor proteins that perform force-generating tasks such as organelle transport and chromosome segregation. It has two identical approximately 960-amino-acid chains containing an amino-terminal globular motor domain, a central alpha-helical region that enables dimer formation through a coiled-coil, and a carboxy-terminal tail domain that binds light chains and possibly an organelle receptor. The kinesin motor domain of approximately 340 amino acids, which can produce movement in vitro, is much smaller than that of myosin (approximately 850 amino acids) and dynein (1,000 amino acids), and is the smallest known molecular motor. Here, we report the crystal structure of the human kinesin motor domain with bound ADP determined to 1.8-A resolution by X-ray crystallography. The motor consists primarily of a single alpha/beta arrowhead-shaped domain with dimensions of 70 x 45 x 45 A. Unexpectedly, it has a striking structural similarity to the core of the catalytic domain of the actin-based motor myosin. Although kinesin and myosin have virtually no amino-acid sequence++ identity, and exhibit distinct enzymatic and motile properties, our results suggest that these two classes of mechanochemical enzymes evolved from a common ancestor and share a similar force-generating strategy.


Temporal stability and correlation of EEG markers and depression questionnaires scores in healthy people.
Mental disorders, especially depression, have become a rising problem in modern society. The development of methods and markers for the early detection of mental disorders is an actual problem. Psychological questionnaires are the only tools for evaluating the symptoms of mental disorders in clinical practice today. The electroencephalography (EEG) based non-invasive and cost-effective method seems feasible for the early detection of depression in occupational and family medicine centers and personal monitoring. The reliability of the EEG markers in the early detection of depression assumes their high temporal stability and correlation with the scores of depression questionnaires. The study was been performed on 17 healthy people over three years. Two hypotheses have been evaluated in the current study: first, the temporal stability of EEG markers is close to the stability of the scores of depression questionnaires, and second, EEG markers and depression questionnaires' scores are not correlated in healthy people. The results of the performed study support both hypotheses: the temporal stability of EEG markers is high and close to the stability of depression questionnaires scores and the correlation between the EEG markers and depression questionnaires scores is not detected in healthy people. The results of the current study contribute to the interpretation of results in depression EEG studies and to the feasibility of EEG markers in the detection of depression.


Parallel and lineage-specific molecular adaptation to climate in boreal black spruce.
In response to selective pressure, adaptation may follow different genetic pathways throughout the natural range of a species due to historical differentiation in standing genetic variation. Using 41 populations of black spruce (Picea mariana), the objectives of this study were to identify adaptive genetic polymorphisms related to temperature and precipitation variation across the transcontinental range of the species, and to evaluate the potential influence of historical events on their geographic distribution. Population structure was first inferred using 50 control nuclear markers. Then, 47 candidate gene SNPs identified in previous genome scans were tested for relationship with climatic factors using an F(ST) -based outlier method and regressions between allele frequencies and climatic variations. Two main intraspecific lineages related to glacial vicariance were detected at the transcontinental scale. Within-lineage analyses of allele frequencies allowed the identification of 23 candidate SNPs significantly related to precipitation and/or temperature variation, among which seven were common to both lineages, eight were specific to the eastern lineage and eight were specific to the western lineage. The implication of these candidate SNPs in adaptive processes was further supported by gene functional annotations. Multiple evidences indicated that the occurrence of lineage-specific adaptive SNPs was better explained by selection acting on historically differentiated gene pools rather than differential selection due to heterogeneity of interacting environmental factors and pleiotropic effects. Taken together, these findings suggest that standing genetic variation of potentially adaptive nature has been modified by historical events, hence affecting the outcome of recent selection and leading to different adaptive routes between intraspecific lineages.


Prospective evaluation of clinical and laboratory effects of intrathecal chemotherapy on children with acute leukemia.
The objective of this prospective 18-month study was to evaluate the clinical and laboratory effects of repeated intrathecal injections of chemotherapy in children with acute leukemia. All procedures were performed under general anesthesia, and complications were prospectively recorded. Laboratory measurements included lumbar puncture opening pressure, cerebrospinal fluid (CSF) chemistry, and cell count and morphology. Central venous pressure and ophthalmologic examinations were also performed. Forty-seven children underwent 247 intrathecal injections of chemotherapy. Adverse effects (13.7% of the procedures) included nausea and vomiting, back pain, and headache. One child each had transient cauda equina syndrome, transient communicating hydrocephalus, and persistent sacral plexus injury. The mean lumbar puncture opening pressure was significantly higher after intrathecal therapy than before (22 +/- 8 vs. 15 +/- 9 cm H2O, P = 0.02) and higher than reported in age-matched children without leukemia. All CSF chemistries, cell count, and morphology were normal. The overall incidence of complications was 13.7%. Most were mild and resolved quickly, but significant neurologic complications did occur. Lumbar puncture opening pressure was significantly higher in children with acute leukemia after intrathecal chemotherapy.


[Participation of forensic medical experts in the examination at the place of occurrence: fundamental legal principles and organizational aspects].
Discussed in this paper are fundamental legal principles and organizational aspects of the participation of forensic medical experts in the examination of the corpses at the place of occurrence. A detailed analysis of the current departmental and sectoral regulations governing the activities of specialists in the field of forensic medicine was performed The analysis demonstrated their positive and negative aspects. These findings were used to develop concrete recommendations for further improvement of these documents.


Allergic contact dermatitis in healthcare workers.
Allergic contact dermatitis is a common occupational skin disease among healthcare workers (HCWs). This study describes causes of allergic contact dermatitis in New Zealand HCWs. All HCWs undergoing patch testing between July 2008 and January 2020 at a public hospital patch test clinic and between June 2019 and January 2020 at a private dermatology clinic were included. Data collected included patient demographics, occupation, results of patch testing and pre- and post-patch test diagnoses. Out of 837 patients tested, 67 were HCWs. The mean age of HCWs was 41 years (SD 14) and 58 (87%) were female. The most common occupations were nurses (40%), allied health (22%) and doctors (18%). Forty-six (69%) patients had a background of atopic dermatitis. Hand dermatitis was the most common presentation (49%), followed by facial/neck dermatitis (25%). Thirty-eight (57%) had at least one positive reaction on patch testing, including 19 (28%) with a positive result of current relevance. There were 16 relevant reactions to rubber accelerators, 11 to fragrances, 10 to preservatives and 6 to corticosteroids. The most frequent relevant allergens were methylisothiazolinone/methylchloroisothiazolinone (4%), hydroxyisohexyl 3-cyclohexene carboxaldehyde (4%) and methylisothiazolinone (4%). The most common allergens in HCWs were rubber chemicals, fragrances, preservatives and topical corticosteroids. Important allergens for patch testing HCWs are outlined, in particular, those with hand dermatitis should be tested to a rubber series.


Factors associated with pre-drinking among nightclub patrons in the city of São Paulo.
The aim of the study was to describe the phenomenon of pre-drinking (alcohol consumption before entering nightclubs or bars) and to identify factors associated with pre-drinking practices among patrons in the city of São Paulo, Brazil. Individual-level data were collected by a portal survey of 2422 patrons at the entrance and at the exit of 31 nightclubs. The nightclubs were selected by two-stage sampling using a probability proportional to the establishments' capacity in the first stage and a systematic sample of patrons in the second stage. Breath alcohol concentration (BrAC) was measured. Face-to-face interview identified pre-drinking characteristics and past-year risk behaviors. Analysis used sample weights to compensate for nightclubs or patrons that were possibly over- or under-represented. Of the study participants, 41.3% (95% confidence interval (CI) = 33.7-49.3) engaged in pre-drinking on the night of the interview. Being male (odds ratio (OR) = 1.98, 95% CI = 1.45-2.71), past-year binge drinking (OR = 2.28, 95% CI = 1.70-3.07), previous episodes of severe effects from drunkenness (OR = 1.77, 95% CI = 1.40-2.22) and sexual risk behavior (OR = 1.67, 95% CI = 1.20-2.33) were associated with recent pre-drinking. Pre-drinking predicted higher BrACs at the nightclub exit. Pre-drinking is prevalent among nightclub patrons and associated with risk behaviors, and is associated with alcohol intoxication at nightclub exits. Environmental prevention strategies must consider pre-drinking as a potential risk factor for alcohol intoxication in nightclubs.


Antiretroviral monotherapy in early stage human immunodeficiency virus disease has no detectable effect on virus load in peripheral blood and lymph nodes.
Initiation of antiretroviral monotherapy early in the course of infection with human immunodeficiency virus may result in a temporary slowing in the rate of disease progression; however, little is known about the virologic effects of early therapy. Virus load was measured in peripheral blood and lymph nodes from 16 antiretroviral-naive patients with a mean CD4 T lymphocyte count of 659 cells/microliter at baseline and after 8 weeks of either no treatment or zidovudine therapy. CD4 T lymphocyte counts and all virologic parameters examined remained unchanged regardless of zidovudine treatment status. Histopathology and virus distribution within lymph nodes remained constant between baseline and week 8 in each patient, indicating that the virologic and histologic parameters examined in a single lymph node are representative of a systemic process. Early antiretroviral monotherapy with zidovudine had no effect on virologic parameters in this group of patients with relatively high CD4 T lymphocyte counts and low measures of virus load at baseline.


[Cell-free system of protein synthesis from rat's cardiac muscle].
A cell-free system of protein synthesis from rat heart muscle is described. The system contained preparations of ribosomes, which were not separated from myofibrillar proteins. The conditions for incorporation of 14C-leucine into the polyribosomes of the fraction were established and its protein synthetizing activity was studied.


Influence of dopaminergic medication on automatic postural responses and balance impairment in Parkinson's disease.
It is still unclear why balance impairment in Parkinson's disease (PD) often responds insufficiently to dopaminergic medication. We have studied this issue in 23 patients with idiopathic PD and 24 healthy controls. Our specific purposes were (a) to investigate the contribution of abnormal automatic postural responses to balance impairment in PD and (b) to assess the influence of dopaminergic medication on abnormal automatic postural responses and balance impairment. Standing subjects received 4 degrees "toe-up" rotational perturbations of a supporting forceplate. We bilaterally recorded posturally destabilizing medium latency (ML) responses from the stretched gastrocnemius muscles and functionally corrective long latency (LL) responses from the shortened tibialis anterior (TA) muscles. We also assessed changes in the center of foot pressure (CFP) and the center of gravity (COG). All patients were tested in the "off" and "on" phases. All controls were tested and retested after 1 h. During the off phase, we found enlarged ML amplitudes and diminished LL amplitudes in patients, together with a markedly increased posterior displacement of the COG. The abnormal ML and LL responses were partially responsible for the increased body sway in patients because the initial forward (destabilizing) displacement of the CFP was increased, while the subsequent backward displacement of the CFP (a measure of the corrective braking action of LL responses) was delayed. Abnormal late automatic or possibly more voluntary postural corrections also contributed substantially to the increased body sway. During the on phase, ML amplitudes were reduced in patients but remained increased compared with controls. LL amplitudes no longer differed between both groups due to a modest, possibly dopamine-related increase in patients and a simultaneous decrease in controls. The abnormal CFP displacement was only partially improved by dopaminergic medication. The later postural corrections were not improved at all. Consequently, the increased posterior COG displacement was not ameliorated during the on phase. We conclude that (a) a combination of abnormal automatic and perhaps more voluntary postural corrections contributes to increased body sway in PD and (b) dopaminergic medication fails to improve balance impairment in PD because early automatic postural responses are only partially corrected, while later occurring postural corrections are not improved at all. These electrophysiological results support clinical observations and suggest that nondopaminergic lesions play a significant role in the pathophysiology of postural abnormalities in PD.


[The capping protein on the slow-growing end of actin filament: erythrocyte tropomodulin].
Erythrocyte tropomodulin (E-Tmod) is first isolated from human erythrocyte membrane as a TM-binding protein. Its N-terminus contains two TM-binding sites and one TM-dependent actin capping domain and C-terminus contains 5 leucine-rich repeats and a TM-independent actin capping domain. As the unique capping protein at the slow-growing end of F-actin, E-Tmod binds to N-terminus of TM and actin and decreases the TM-coated F-actin depolymerization. E-Tmod encoding gene is highly conserved and E-Tmod is distributed widely in erythrocytes and cardiomyocytes, etc. E-Tmod plays a pivotal role in organizing F-actin and cytoskeleton and maintaining the mechanical properties of the cells.


Reconstitution of maltose chemotaxis in Escherichia coli by addition of maltose-binding protein to calcium-treated cells of maltose regulon mutants.
Maltose chemotaxis was reconstituted in delta malE cells lacking maltose-binding protein (MBP). Purified MBP was introduced into intact cells during incubation with 250 mM CaCl2 in Tris-hydrochloride buffer at 0 degrees C. After removal of extracellular CaCl2 and MBP, chemotaxis was measured with tethered bacteria in a flow chamber or with free-swimming cells in a capillary assay. About 20% of tethered cells responded to 10(-4) M maltose; the mean response times were about half those of CaCl2-treated wild-type cells (100 s as opposed to 190 s). In capillary tests, the maltose response of reconstituted cells was between 15 and 40% of the aspartate response, about the same percentage as in wild-type cells. The best reconstitution was seen with 0.5 to 1 mM MBP in the reconstitution mixture, which is similar to the periplasmic MBP concentration estimated for maltose-induced wild-type cells. Strains containing large deletions of the malB region and malT mutants lacking the positive regulator gene of the mal regulon also could be reconstituted for maltose chemotaxis, showing that no product of the mal regulon other than MBP is essential for maltose chemotaxis.


Identification and Characterization of a Novel Nontranslated Sequence Variant of the Human Intestinal Di-/Tripeptide Transporter, hPEPT1.
The human H(+)-coupled di-/tripeptide transporter (hPEPT1) mediates intestinal absorption of dietary di- and tripeptides, as well as several peptidomimetic drug compounds. The aim of the present study was to investigate the possible role of the hPEPT1 variant hPEPT1-RF in hPEPT1 regulation. However, the proposed hPEPT1-RF mRNA sequence could not be detected in Caco-2 cells or in human intestinal samples. Instead, a new sequence variant, hPEPT1-RFI, was found, which is almost identical to the proposed hPEPT1-RF, except for two nucleotide insertions and one deletion that resulted in a changed open reading frame as compared to hPEPT1-RF. In vitro translation analysis showed that hPEPT1-RFI was not translated. In conclusion, the existence of hPEPT1-RF could not be confirmed; furthermore, the identified sequence variant, hPEPT1-RFI, does not appear to be translated and is therefore unlikely to have a regulatory effect on hPEPT1 transport activity.