Mental health treatment expenditure trends, 1986-2003.
This study determined spending on mental health treatment in the United States over time by provider and payer relative to all health spending. Estimates were developed to be consistent with the National Health Expenditure Accounts. Numerous public data sources were used. Mental health treatment expenditures grew from $33 billion in 1986 to $100 billion in 2003. In real 2003 dollars, spending per capita on mental health treatment rose from $205 to $345. The average annual nominal total mental health growth rate was 6.7%. In comparison, total health care expenditures increased by 8.0%. As a result of the slower growth rate of mental health expenditures compared with all health spending, mental health fell from 8% of all health expenditures in 1986 to 6% in 2003. Total national health spending increased by approximately $1.175 trillion from 1986 to 2003; of this, 6% is attributed to an increase in mental health spending. The mix of services has changed, with more care being provided through prescription drugs and in outpatient settings and less in inpatient settings. Payer mix has also shifted, with Medicaid taking a more prominent role. Spending on mental health treatment has increased over the past decade, reflecting increases in the number of individuals receiving mental health treatment, particularly prescription drugs and outpatient treatment. Changes in payer and provider mix raise new challenges for ensuring quality and access.


Homozygous hypobetalipoproteinemia: a disease distinct from abetalipoproproteinemia at the molecular level.
apoB DNA, RNA, and protein from two patients with homozygous hypobetalipoproteinemia (HBL) were evaluated and compared with normal individuals. Southern blot analysis with 10 different cDNA probes revealed a normal gene without major insertions, deletions, or rearrangements. Northern and slot blot analyses of total liver mRNA from HBL patients documented a normal size apoB mRNA that was present in greatly reduced quantities. ApoB protein was detected within HBL hepatocytes utilizing immunohistochemical techniques; however, it was markedly reduced in quantity when compared with control samples. No apoB was detectable in the plasma of HBL individuals with an ELISA assay. These data are most consistent with a mutation in the coding portion of the apoB gene in HBL patients, leading to an abnormal apoB protein and apoB mRNA instability. These results are distinct from those previously noted in abetalipoproteinemia, which was characterized by an elevated level of hepatic apoB mRNA and accumulation of intracellular hepatic apoB protein.


Economic Costs Attributable to Diabetes in Each U.S. State.
To estimate direct medical and indirect costs attributable to diabetes in each U.S. state in total and per person with diabetes. We used an attributable fraction approach to estimate direct medical costs using data from the 2013 State Health Expenditure Accounts, 2013 Behavioral Risk Factor Surveillance System, and the Centers for Medicare & Medicaid Services' 2013-2014 Minimum Data Set. We used a human capital approach to estimate indirect costs measured by lost productivity from morbidity (absenteeism, presenteeism, lost household productivity, and inability to work) and premature mortality, using the 2008-2013 National Health Interview Survey, 2013 daily housework value data, 2013 mortality data from the Centers for Disease Control and Prevention Wide-ranging Online Data for Epidemiologic Research, and mean wages from the 2014 Bureau of Labor Statistics. Costs were adjusted to 2017 U.S. dollars. The estimated median state economic cost was $5.9 billion, ranging from $694 million to $55.5 billion, in total and $18,248, ranging from $15,418 to $30,915, per person with diabetes. The corresponding estimates for direct medical costs were $2.8 billion (range $0.3-22.9) and $8,544 (range $6,591-12,953) and for indirect costs were $3.0 billion (range $0.4-32.6) and $9,672 (range $7,133-17,962). In general, the estimated state median indirect costs resulting from morbidity were larger than costs from mortality both in total and per person with diabetes. Economic costs attributable to diabetes were large and varied widely across states. Our comprehensive state-specific estimates provide essential information needed by state policymakers to monitor the economic burden of the disease and to better plan and evaluate interventions for preventing type 2 diabetes and managing diabetes in their states.


Reconstructing Roma history from genome-wide data.
The Roma people, living throughout Europe and West Asia, are a diverse population linked by the Romani language and culture. Previous linguistic and genetic studies have suggested that the Roma migrated into Europe from South Asia about 1,000-1,500 years ago. Genetic inferences about Roma history have mostly focused on the Y chromosome and mitochondrial DNA. To explore what additional information can be learned from genome-wide data, we analyzed data from six Roma groups that we genotyped at hundreds of thousands of single nucleotide polymorphisms (SNPs). We estimate that the Roma harbor about 80% West Eurasian ancestry-derived from a combination of European and South Asian sources-and that the date of admixture of South Asian and European ancestry was about 850 years before present. We provide evidence for Eastern Europe being a major source of European ancestry, and North-west India being a major source of the South Asian ancestry in the Roma. By computing allele sharing as a measure of linkage disequilibrium, we estimate that the migration of Roma out of the Indian subcontinent was accompanied by a severe founder event, which appears to have been followed by a major demographic expansion after the arrival in Europe.


Escitalopram in the treatment of impulsive-compulsive internet usage disorder: an open-label trial followed by a double-blind discontinuation phase.
Isolated reports suggest that escitalopram may be effective for impulsive-compulsive Internet usage disorder (IC-IUD), an impulse-control disorder characterized by excessive time spent on the Internet at the expense of occupational, relationship, and social activities. To assess the safety and efficacy of escitalopram in IC-IUD, we conducted a 10-week, open-label trial followed by a 9-week, double-blind, placebo-controlled discontinuation phase. From December 2002 to October 2004, 19 adult subjects with IC-IUD (defined as time consuming, uncontrollable, distressing, and resulting in social, occupational, or financial difficulties) were enrolled. Escitalopram was started at 10 mg/day, then increased and maintained at 20 mg/day for 10 weeks at the end of which completers were randomly assigned to placebo or escitalopram for 9 additional weeks. Two key outcome measures were used: hours spent weekly in nonessential Internet use and overall clinical response (subjects rated "much improved" or "very much improved" on the Clinical Global Impressions-Improvement scale [CGI-I]). Fourteen subjects completed the entire study. At the end of the 10th week of open-label esci-talopram, Internet usage decreased significantly from a mean of 36.8 hours/week at baseline to 16.5 hours/week (paired t test: t = 3.58; p = .002). In addition, 64.7% of the sample (N = 11) were considered CGI-I responders. At the end of the double-blind phase, there were no significant differences in outcome measures between patients taking placebo compared to escitalopram (analysis of variance with repeated measures, p > .05). Patients showed a significant improvement of IC-IUD symptoms during the open-label escitalopram phase. There was no significant difference between the escitalopram and placebo groups at the end of the subsequent double-blind phase; both groups maintained the gains made in the initial open-label treatment. Larger controlled trials are needed to investigate the efficacy of this and other pharmacologic agents in the treatment of IC-IUD. clinicaltrials.gov Identifier: NCT00565422.


Incontinence-Associated Dermatitis, Characteristics and Relationship to Pressure Injury: A Multisite Epidemiologic Analysis.
The purpose of this study was to measure the prevalence of incontinence-associated dermatitis (IAD) among incontinent persons in the acute care setting, characteristics of IAD in this group, and associations among IAD, urinary, fecal, and dual incontinence, immobility, and pressure injury in the sacral area. Descriptive and correlational analysis of data from a large database of IAD, and pressure injuries of sacral area and heels. The sample comprised 5342 adult patients in acute care facilities in 36 states representing all regions of the United States. Facilities used a variety of products for prevention of IAD and sacral area pressure injuries. Data were collected for use in a national quality improvement study evaluating current practices related to the prevention of IAD and pressure injuries affecting the sacral area and heels. Data were exported to a spreadsheet, and triple checked for accuracy before being imported to a statistical analysis software program. Descriptive statistics were used to describe prevalence rates for incontinence, types of incontinence, IAD, characteristics of IAD, and pressure injuries. Multivariate logistic regression analysis was conducted on the end point of facility-acquired sacral/buttock pressure injury and the risk factors of immobility and type of incontinence. More than one-third of patients (n = 2492 of 5342 patients; 46.6%) were incontinent of urine, stool, or both. The overall prevalence rate of IAD was 21.3% (1140/5342); the prevalence of IAD among patients with incontinence was 45.7% (1140/2492). Slightly more than half of the IAD was categorized as mild (596/1140, 52.3%), 27.9% (318/1140) was categorized as moderate, and 9.2% (105/1140) was deemed severe. In addition, 14.8% (169/1140) of patients with IAD also had a fungal rash. The prevalence of pressure injury in the sacral area among individuals with incontinence was 17.1% (427/2492), and the prevalence of full-thickness pressure injury in this population was 3.8% (95/2492). Multivariate analysis revealed that both presence of IAD (odds ratio [OR], 4.56; 95% confidence interval [CI], 3.68-5.65) and immobility (OR, 3.56; 95% CI, 2.73-4.63) was associated with a significantly increased likelihood of developing a sacral pressure injury. Multivariate analysis also revealed that presence of IAD (OR, 2.65; 95% CI, 1.74-4.03) and immobility (OR, 6.05; 95% CI, 3.14-11.64) was associated with a significantly increased likelihood of developing full-thickness sacral pressure injury. Our study findings are consistent with prior research, supporting a clinically relevant association between IAD and pressure injury of the sacral area. This risk persists even after controlling for presence of immobility, suggesting that IAD functions as an independent risk factor for pressure injury occurrence.


Effects of pressure on the skin exerted by clothing on responses of urinary catecholamines and cortisol, heart rate and nocturnal urinary melatonin in humans.
The study investigated how the pressure exerted on the skin by clothing worn while working in the daytime affected the urinary excretion of adrenaline, noradrenaline and cortisol, heart rate, and also melatonin secretion at night. Nine young women (experiment I) and seven young women (experiment II) participated. Participants wore either a 100% cotton jacket (tight clothes, TC) or a 100% cotton T-shirt (loose clothes, LC). Loose-fitting, 100% cotton tank tops and panties were worn as underwear in both the TC and the LC groups. The main results can be summarized as follows: (1) urinary excretion of adrenaline, noradrenaline and cortisol was facilitated, and the amounts of urinary excretion were significantly higher when TC were worn. Heart rate was significantly higher in the TC group; (2) nocturnal urinary melatonin excretion was significantly greater in the TC group. These results are discussed in terms of an enhancement of diurnal sympathetic nervous system activity caused by pressure on the skin produced by tight clothing.


Towards positive diagnosis of the irritable bowel.
A questionnaire to establish the presence of 15 symptoms thought to be typical of the irritable bowel syndrome (IBS) was given to 109 unselected patients referred to gastroenterology or surgery clinics with abdominal pain or a change in bowel habit or both. Review of case records 17--26 months later established a definite diagnosis of IBS in 32 patients and of organic disease in 33. Four symptoms were significantly more common among patients with IBS--namely, distension, relief of pain with bowel movement, and looser and more frequent bowel movements with the onset of pain. Mucus and a sensation of incomplete evacuation were also common in these patients. The more of these symptoms that were present the more likely was it that the patient's pain or altered bowel habit, or both, was due to IBS. We conclude that a careful history can increase diagnostic confidence and reduce the amount of investigation in many patients with chronic abdominal pain.


Posttraumatic stress and co-morbidity following myocardial infarction among older patients: the role of coping.
This study aimed to investigate co-morbidity and coping strategies among older patients who suffer from different levels of posttraumatic stress disorder (PTSD) following myocardial infarction. Ninety-six older myocardial infarction (MI) patients were recruited from two general practices and completed the Posttraumatic Stress Diagnostic Scale (PDS), the General Health Questionnaire-28 (GHQ-28) and the COPE Scale. Ninety-two older patients with no previous MI experience constituted the control. Using the PDS, 30, 42 and 28% had full, partial and no-PTSD respectively. There were significant differences between the patient groups and the control on all GHQ-28 sub-scales. Significant differences were also identified between the patient groups in the following coping strategies: seeking emotional social support, suppression of competing activities, restraint coping, focusing on and venting of emotion, mental and behavioural disengagement. Controlling for bypass surgery, previous mental health difficulties, angioplasty, heart failure and angina, MANCOVA results did not change the overall results of the GHQ-28 but changed the results of coping in that seeking emotional social support and behavioural disengagement stopped being significant. Coping was a partial mediator between different levels of post-MI PTSD and co-morbidity. Depending on the severity of PTSD symptoms, co-morbidity and coping strategies can vary among older patients. Older patients with full-PTSD tend to use both maladaptive coping strategies as well as problem-focused coping.


Is surgical spinal decompression for supratentorial GBM symptomatic drop down metastasis warranted? A case report and review of literature.
Symptomatic spinal metastasis from an intracranial primary glioblastoma multiforme (GBM) is very rare. Our literature search identified a total of 42 such patients of which 11 were treated with surgical decompression for spinal metastasis with only one such report from the pediatric age group. Previous studies have reported variable outcomes after surgical management. We report the case of a 16-year-old boy who underwent surgical spinal decompression for spinal metastasis after intracranial GBM. The patient regained motor and autonomic function following surgery and reported improvement in pain. We also present findings from a literature review using the PubMed database from 1985 to June 2013 on this subject and compare radiation therapy with surgical decompression as palliative modalities in such patients. There are no evidence-based guidelines available on the subject and no treatment regimen has yet demonstrated survival benefit in these patients. Surgical decompression may be a better option for patients with focal resectable lesions and who are medically stable to tolerate the procedure.


Complete unfolding of the titin molecule under external force.
Titin (also known as connectin) is a giant filamentous protein that spans the distance between the Z- and M-lines of the vertebrate muscle sarcomere. Several earlier studies have implicated titin as playing a fundamental role in maintaining sarcomeric structural integrity and generating the passive force of muscle. The elastic properties of titin were characterized in recent single-molecule mechanical works that described the molecule as an entropic spring in which partial unfolding may take place at high forces during stretch and refolding at low forces during release. In the present work titin molecules were stretched using a laser tweezer with forces above 400 pN. The high external forces resulted in complete mechanical unfolding of the molecule, characterized by the disappearance of force hysteresis at high forces. Titin refolded following complete denaturation, as the hysteresis at low forces reappeared in subsequent stretch-release cycles. The broad force range throughout which unfolding occurred indicates that the various globular domains in titin require different unfolding forces due to differences in the activation energies for their unfolding.


Oral NaHCO3 Activates a Splenic Anti-Inflammatory Pathway: Evidence That Cholinergic Signals Are Transmitted via Mesothelial Cells.
We tested the hypothesis that oral NaHCO3 intake stimulates splenic anti-inflammatory pathways. Following oral NaHCO3 loading, macrophage polarization was shifted from predominantly M1 (inflammatory) to M2 (regulatory) phenotypes, and FOXP3+CD4+ T-lymphocytes increased in the spleen, blood, and kidneys of rats. Similar anti-inflammatory changes in macrophage polarization were observed in the blood of human subjects following NaHCO3 ingestion. Surprisingly, we found that gentle manipulation to visualize the spleen at midline during surgical laparotomy (sham splenectomy) was sufficient to abolish the response in rats and resulted in hypertrophy/hyperplasia of the capsular mesothelial cells. Thin collagenous connections lined by mesothelial cells were found to connect to the capsular mesothelium. Mesothelial cells in these connections stained positive for the pan-neuronal marker PGP9.5 and acetylcholine esterase and contained many ultrastructural elements, which visually resembled neuronal structures. Both disruption of the fragile mesothelial connections or transection of the vagal nerves resulted in the loss of capsular mesothelial acetylcholine esterase staining and reduced splenic mass. Our data indicate that oral NaHCO3 activates a splenic anti-inflammatory pathway and provides evidence that the signals that mediate this response are transmitted to the spleen via a novel neuronal-like function of mesothelial cells.


The EMBRACE web service collection.
The EMBRACE (European Model for Bioinformatics Research and Community Education) web service collection is the culmination of a 5-year project that set out to investigate issues involved in developing and deploying web services for use in the life sciences. The project concluded that in order for web services to achieve widespread adoption, standards must be defined for the choice of web service technology, for semantically annotating both service function and the data exchanged, and a mechanism for discovering services must be provided. Building on this, the project developed: EDAM, an ontology for describing life science web services; BioXSD, a schema for exchanging data between services; and a centralized registry (http://www.embraceregistry.net) that collects together around 1000 services developed by the consortium partners. This article presents the current status of the collection and its associated recommendations and standards definitions.


A study of temperature dependent local atomic displacements in a Ba(Fe(1-x)Co(x))2As2 superconductor.
We have studied the local structure of a Ba(Fe(1-x)Co(x))2As2 superconductor using temperature dependent extended X-ray absorption fine structure (EXAFS) measurements. Polarized EXAFS at the Fe K-edge on an optimally doped (x = 0.06) single crystal has permitted us to determine atomic displacements across the superconducting transition temperature (T(c)). The Fe-As bondlength hardly shows any change with temperature; however, the Fe-Fe sublattice reveals a sharp anomaly across T(c), indicated by a significant drop in mean square relative displacements, similar to the one known for cuprates and A15-type superconductors. We have also found a large atomic disorder around the substituted Co, revealed by polarized Co K-edge EXAFS measurements. The Co-Fe/Co bonds are more flexible than the Fe-Fe bonds with the As-height in Co-containing tetrahedra being larger than the one in FeAs4. The results suggest that the local Fe-Fe bondlength fluctuations and the atomic disorder in this sub-lattice should have some important role in the superconductivity of Ba(Fe(1-x)Co(x))2As2 pnictides.


The burden of road traffic injuries in Nigeria: results of a population-based survey.
Mortality from road traffic injuries in sub-Saharan Africa is among the highest in the world, yet data from the region are sparse. To date, no multi-site population-based survey on road traffic injuries has been reported from Nigeria, the most populated country in Africa. To explore the epidemiology of road traffic injury in Nigeria and provide data on the populations affected and risk factors for road traffic injury. Data from a population-based survey using two-stage stratified cluster sampling. SUBJECTS/ SETTING: Road traffic injury status and demographic information were collected on 3082 respondents living in 553 households in seven of Nigeria's 37 states. Incidence rates were estimated with confidence intervals based on a Poisson distribution; Poisson regression analysis was used to calculate relative risks for associated factors. The overall road traffic injury rate was 41 per 1000 population (95% CI 34 to 49), and mortality from road traffic injuries was 1.6 per 1000 population (95% CI 0.5 to 3.8). Motorcycle crashes accounted for 54% of all road traffic injuries. The road traffic injury rates found for rural and urban respondents were not significantly different. Increased risk of injury was associated with male gender among those aged 18-44 years, with a relative risk of 2.96 when compared with women in the same age range (95% CI 1.72 to 5.09, p<0.001). The road traffic injury rates found in this survey highlight a neglected public health problem in Nigeria. Simple extrapolations from this survey suggest that over 4 million people may be injured and as many as 200 000 potentially killed as the result of road traffic crashes annually in Nigeria. Appropriate interventions in both the health and transport sectors are needed to address this significant cause of morbidity and mortality in Nigeria.


Comparison of spirometric thresholds in diagnosing smoking-related airflow obstruction.
Diagnosis of chronic obstructive pulmonary disease is based on detection of airflow obstruction on spirometry. There is no consensus regarding using a fixed threshold to define airflow obstruction versus using the lower limit of normal (LLN) adjusted for age. We compared the accuracy and discrimination of the Global Initiative for Chronic Obstructive Lung Disease (GOLD) recommended fixed ratio of forced expiratory volume in the first second/forced vital capacity<0.70 with LLN in diagnosing smoking-related airflow obstruction using CT-defined emphysema and gas trapping as the disease gold standard. Data from a large multicentre study (COPDGene), which included current and former smokers (age range 45-80 years) with and without airflow obstruction, were analysed. Concordance between spirometric thresholds was measured. The accuracy of the thresholds in diagnosing emphysema and gas trapping was assessed using quantitative CT as gold standard. 7743 subjects were included. There was very good agreement between the two spirometric cutoffs (κ=0.85; 95% CI 0.83 to 0.86, p<0.001). 7.3% were discordant. Subjects with airflow obstruction by fixed ratio only had a greater degree of emphysema (4.1% versus 1.2%, p<0.001) and gas trapping (19.8% vs 7.5%, p<0.001) than those positive by LLN only, and also smoking controls without airflow obstruction (4.1% vs 1.9% and 19.8% vs 10.9%, respectively, p<0.001). On follow-up, the fixed ratio only group had more exacerbations than smoking controls. Compared with the fixed ratio, the use of LLN fails to identify a number of patients with significant pulmonary pathology and respiratory morbidity.


Water safety plan enhancements with improved drinking water quality detection techniques.
Drinking water quality has been regulated in most European countries for nearly two decades by the drinking water directive 98/83/EC. The directive is now under revision with the goal of meeting stricter demands for safe water for all citizens, as safe water has been recognized as a human right by the United Nations. An important change to the directive is the implementation of a risk-based approach in all regulated water supplies. The European Union Framework Seventh Programme Aquavalens project has developed several new detection technologies for pathogens and indicators and tested them in water supplies in seven European countries. One of the tasks of the project was to evaluate the impact of these new techniques on water safety and on water safety management. Data were collected on risk factors to water safety for five large supplies in Denmark, Germany, Spain and the UK, and for fifteen small water supplies in Scotland, Portugal and Serbia, via a questionnaire aiming to ascertain risk factors and the stage of implementation of Water Safety Plans, and via site-specific surveys known as Sanitary Site Inspection. Samples were collected from the water supplies from all stages of water production to delivery. Pathogens were detected in around 23% of the 470 samples tested. Fecal contamination was high in raw water and even in treated water at the small supplies. Old infrastructure was considered a challenge at all the water supplies. The results showed that some of the technique, if implemented as part of the water safety management, can detect rapidly the most common waterborne pathogens and fecal pollution indicators and therefore have a great early warning potential; can improve water safety for the consumer; can validate whether mitigation methods are working as intended; and can confirm the quality of the water at source and at the tap.


Sensitivity of the N-methyl-D-aspartate receptor to polyamines is controlled by NR2 subunits.
The endogenous polyamine spermine has multiple effects on the N-methyl-D-aspartate (NMDA) receptor. These include an increase in the magnitude of NMDA-induced whole-cell currents that is seen in the presence of saturating concentrations of glycine ("glycine-independent" stimulation), an increase in the affinity of the receptor for glycine ("glycine-dependent" stimulation), and voltage-dependent inhibition. Although many of the properties of native NMDA receptors are seen with homomeric NR1 receptors expressed in Xenopus oocytes, we have found that the effects of spermine are differentially regulated by NR2 subunits in heteromeric NR1/NR2 receptors. Glycine-independent stimulation by spermine occurred at homomeric NR1A receptors, which lack the amino-terminal insert in NR1, and at heteromeric NR1A/NR2B receptors but not at heteromeric NR1A/NR2A or NR1A/NR2C receptors. Glycine-independent stimulation was not seen at homomeric NR1B receptors, which include the amino-terminal insert in NR1, or at heteromeric receptors containing NR1B. Thus, glycine-independent stimulation by polyamines requires the presence of an NR1 variant, such as NR1A, that lacks the amino-terminal insert, but the manifestation of the stimulatory effect is controlled by the type of NR2 subunit present in a heteromeric complex. Glycine-dependent stimulation was seen at NR1A/NR2A and NR1A/NR2B receptors and may therefore involve a second polyamine binding site distinct from that which produces glycine-independent stimulation. The voltage-dependent inhibitory effect of spermine, which is more pronounced at hyperpolarized membrane potentials, occurred with similar magnitudes at NR1A/NR2A and NR1A/NR2B receptors but was absent at NR1A/NR2C receptors. Thus, NR2 subunits control both the stimulatory and inhibitory effects of spermine at NMDA receptors. Stimulation but not inhibition by spermine was seen at NR1A/NR2B receptors in the presence of extracellular Mg2+. Stimulation, seen in the presence of physiological concentrations of Ca2+ and Mg2+, may be the predominant effect of polyamines at NMDA receptors in the intact nervous system.


Antagonism of L-type Ca2+ channels CaV1.3 and CaV1.2 by 1,4-dihydropyrimidines and 4H-pyrans as dihydropyridine mimics.
The L-type calcium channel (LTCC) CaV1.3 is regarded as a new potential therapeutic target for Parkinson's disease. Calcium influx through CaV1.3 LTCC during autonomous pacemaking in adult dopaminergic neurons of the substantia nigra pars compacta is related to the generation of mitochondrial oxidative stress in animal models. Development of a CaV1.3 antagonist selective over CaV1.2 is essential because CaV1.2 pore-forming subunits are the predominant form of LTCCs and are abundant in the central nervous and cardiovascular systems. We have explored 1,4-dihydropyrimidines and 4H-pyrans to identify potent and selective antagonists of CaV1.3 relative to CaV1.2 LTCCs. A library of 36 dihydropyridine (DHP)-mimic 1,4-dihydropyrimidines and 4H-pyrans was synthesized, and promising chiral compounds were resolved. The antagonism studies of CaV1.3 and CaV1.2 LTCCs using DHP mimic compounds showed that dihydropyrimidines and 4H-pyrans are effective antagonists of DHPs for CaV1.3 LTCCs. Some 1,4-dihydropyrimidines are more selective than isradipine for CaV1.3 over CaV1.2, shown here by both calcium flux and patch-clamp electrophysiology experiments, where the ratio of antagonism is around 2-3. These results support the hypothesis that the modified hydrogen bonding donor/acceptors in DHP-mimic dihydropyrimidines and 4H-pyrans can interact differently with DHP binding sites, but, in addition, the data suggest that the binding sites of DHP in CaV1.3 and CaV1.2 LTCCs are very similar.


Bilateral prophylactic oophorectomy and bilateral prophylactic mastectomy in a prospective cohort of unaffected BRCA1 and BRCA2 mutation carriers.
Women with BRCA1 or BRCA2 (BRCA1/2) mutations can reduce cancer incidence and mortality by using bilateral prophylactic oophorectomy (BPO) or bilateral prophylactic mastectomy (BPM). The availability of these risk-reduction strategies is an important consideration in the decision to undergo genetic testing. We evaluated the use of BPO and BPM in a prospective sample of 537 female BRCA1/2 mutation carriers from 17 centers in North America and Europe. These women were aged > 30 years, had no BPM, BPO, breast cancer, or ovarian cancer before the disclosure of their genetic test results and were followed for > or = 6 months. Bilateral prophylactic oophorectomy is used significantly more frequently than BPM (55% vs. 21%; P < .001). Bilateral prophylactic oophorectomy was more common among women age > or = 40 years compared with women aged < 40 years (68% vs. 43%; P < .001) and among parous women compared with nulliparous women (60% vs. 39%; P < .001). There was no difference in BPM (P = .83) or BPO (P = .09) in BRCA1 versus BRCA2 carriers. Multivariate models identified age and parity as a predictor of BPO in BRCA1 carriers; age and ovarian cancer family history in BRCA2 carriers; parity and ovarian cancer family history as a predictor of BPM in BRCA1 carriers; and smoking and ovarian cancer family history in BRCA2 carriers. Bilateral prophylactic oophorectomy is more commonly used than BPM in unaffected BRCA1/2 mutation carriers. Parity, age, and family history can also influence BPO and BPM uptake. Consistent with current recommendations, BPO is used by the majority of parous women aged > 40 years.


Impaired pregnenolone secretion after combined cyproterone acetate and ethynyl estradiol therapy in hirsute patients.
6 women affected by hirsutism, either of idiopathic origin or due to polycystic ovary syndrome, have been treated with cyproterone acetate and ethynyl estradiol in combined therapy using, respectively, 100 mg and 50 micrograms/day, from the 5th to the 25th day of the cycle. The adrenal function was assessed before treatment and at the end of the 4th month of therapy, evaluating the peripheral plasma concentrations of pregnenolone (delta 5P), progesterone, 17-OH-progesterone, dehydro-epiandrosterone sulfate, androstenedione, testosterone, and cortisol in basal conditions and after dexamethasone suppression and an adrenocorticotropic hormone (ACTH) stimulation test. A group of healthy, untreated females were examined in the early follicular phase, as controls, Before therapy, the hirsute patient showed testosterone and androstenedione plasma levels, which were significantly higher than in the controls, and a significant reduction in pregnenolone response to ACTH. After 4 months of therapy with cyproterone acetate plus ethynyl estradiol, a significant decrease was found in testosterone and androstenedione plasma levels, and pregnenolone basal plasma levels, dexamethasone suppressibility, and response to ACTH were also markedly reduced, showing a significant difference versus the same patients before therapy and versus the control group. The existence of an impairment in adrenal function after cyproterone acetate plus ethynyl estradiol therapy at the given dose seems to be evident only in the case of directly ACTH-dependent adrenal enzymatic activities responsible for cholesterol cleavage to pregnenolone.


Essential oil of Siparuna guianensis as an alternative tool for improved lepidopteran control and resistance management practices.
Although the cultivation of transgenic plants expressing toxins of Bacillus thuringiensis (Bt) represents a successful pest management strategy, the rapid evolution of resistance to Bt plants in several lepidopteran pests has threatened the sustainability of this practice. By exhibiting a favorable safety profile and allowing integration with pest management initiatives, plant essential oils have become relevant pest control alternatives. Here, we assessed the potential of essential oils extracted from a Neotropical plant, Siparuna guianensis Aublet, for improving the control and resistance management of key lepidopteran pests (i.e., Spodoptera frugiperda and Anticarsia gemmatalis). The essential oil exhibited high toxicity against both lepidopteran pest species (including an S. frugiperda strain resistant to Cry1A.105 and Cry2Ab Bt toxins). This high insecticidal activity was associated with necrotic and apoptotic effects revealed by in vitro assays with lepidopteran (but not human) cell lines. Furthermore, deficits in reproduction (e.g., egg-laying deterrence and decreased egg viability), larval development (e.g., feeding inhibition) and locomotion (e.g., individual and grouped larvae walking activities) were recorded for lepidopterans sublethally exposed to the essential oil. Thus, by similarly and efficiently controlling lepidopteran strains susceptible and resistant to Bt toxins, the S. guianensis essential oil represents a promising management tool against key lepidopteran pests.


Caregivers' Experiences of Pathways to Care for Seriously Ill Children in Cape Town, South Africa: A Qualitative Investigation.
Understanding caregivers' experiences of care can identify barriers to timely and good quality care, and support the improvement of services. We aimed to explore caregivers' experiences and perceptions of pathways to care, from first access through various levels of health service, for seriously ill and injured children in Cape Town, South Africa, in order to identify areas for improvement. Semi-structured, qualitative interviews were conducted with primary caregivers of children who were admitted to paediatric intensive care or died in the health system prior to intensive care admission. Interviews explored caregivers' experiences from when their child first became ill, through each level of health care to paediatric intensive care or death. A maximum variation sample of transcripts was purposively sampled from a larger cohort study based on demographic characteristics, child diagnosis, and outcome at 30 days; and analysed using the method of constant comparison. Of the 282 caregivers who were interviewed in the larger cohort study, 45 interviews were included in this qualitative analysis. Some caregivers employed 'tactics' to gain quicker access to care, including bypassing lower levels of care, and negotiating or demanding to see a healthcare professional ahead of other patients. It was sometimes unclear how to access emergency care within facilities; and non-medical personnel informally judged illness severity and helped or hindered quicker access. Caregivers commonly misconceived ambulances to be slow to arrive, and were concerned when ambulance transfers were seemingly not prioritised by illness severity. Communication was often good, but some caregivers experienced language difficulties and/or criticism. Interventions to improve child health care could be based on: reorganising the reception of seriously ill children and making the emergency route within healthcare facilities clear; promoting caregivers' use of ambulances and prioritising transfers according to illness severity; addressing language barriers, and emphasising the importance of effective communication to healthcare providers.


Germinal Center Selection and Affinity Maturation Require Dynamic Regulation of mTORC1 Kinase.
During antibody affinity maturation, germinal center (GC) B cells cycle between affinity-driven selection in the light zone (LZ) and proliferation and somatic hypermutation in the dark zone (DZ). Although selection of GC B cells is triggered by antigen-dependent signals delivered in the LZ, DZ proliferation occurs in the absence of such signals. We show that positive selection triggered by T cell help activates the mechanistic target of rapamycin complex 1 (mTORC1), which promotes the anabolic program that supports DZ proliferation. Blocking mTORC1 prior to growth prevented clonal expansion, whereas blockade after cells reached peak size had little to no effect. Conversely, constitutively active mTORC1 led to DZ enrichment but loss of competitiveness and impaired affinity maturation. Thus, mTORC1 activation is required for fueling B cells prior to DZ proliferation rather than for allowing cell-cycle progression itself and must be regulated dynamically during cyclic re-entry to ensure efficient affinity-based selection.


Ambient Volatile Organic Compounds and Racial/Ethnic Disparities in Gestational Diabetes Mellitus: Are Asian/Pacific Islander Women at Greater Risk?
US Asian/Pacific Islander (API) communities experience high air pollution levels. APIs may be predisposed to pancreatic β-cell dysfunction and have the highest prevalence of gestational diabetes mellitus (GDM) compared with other racial/ethnic groups. Exposure to high levels of volatile organic compounds (VOCs) impairs pancreatic β-cell function, leading to insulin resistance, but racial/ethnic differences in this association are unexamined. We analyzed singleton deliveries (n = 220,065) from the Consortium on Safe Labor (2002-2008). Exposure to 14 VOCs in each hospital referral region was based on modified Community Multiscale Air Quality models. Logistic regression estimated odds ratios for GDM associated with high (≥75th percentile) versus low (<75th percentile) VOC exposure 3 months before conception and during the first trimester of pregnancy. Preconception and first-trimester exposure to high VOC levels was associated with increased odds of GDM among whites and APIs. GDM risk was significantly higher for APIs than whites for most VOCs. Preconception benzene exposure was associated with 29% (95% confidence interval: 12, 47) increased odds of GDM among whites compared with 45% (95% confidence interval: 16, 81) increased odds among APIs. These findings highlight environmental health disparities affecting pregnant women. Increased focus on GDM risk in US API communities is warranted.


Influence of loop shortening on the metal binding site of cupredoxin pseudoazurin.
Atomic resolution structures of the pseudoazurin (PAZ) variant into which the shorter ligand-containing loop of amicyanin (AMI) is introduced have been determined. The mutated loop adopts a different conformation in PAZAMI than in AMI. The copper site structure is affected, with the major influence being an increased axial interaction resulting in the shortest Cu(II)-S(Met) bond observed for the cupredoxin family of electron-transfer proteins. This is accompanied by a lengthening of the important Cu-S(Cys) bond and enhanced tetragonal distortion, consistent with the influence of the PAZAMI loop contraction on the UV/vis spectrum. The change in active site geometry is the major cause of the 50 mV decrease in reduction potential. The copper site structure changes very little upon reduction, consistent with the distorted site still possessing the properties required to facilitate rapid electron transfer. The exposed His ligand on the loop protonates in the reduced protein and reasons for the increased pKa compared to that of PAZ are discussed. The area surrounding the His ligand is more hydrophobic in PAZAMI than in PAZ, while electron self-exchange, which involves homodimer formation via this surface patch, is decreased. The nature of the side chains in this region, as dictated by the sequence of the ligand-containing loop, is a more significant factor than hydrophobicity for facilitating transient protein interactions in PAZ. The structure of PAZAMI demonstrates the importance of loop-scaffold interactions for metal sites in proteins.


The human qualities needed to complete the global eradication of polio.
Although the 99% decrease seen in global polio incidence between 1988 and 2000 represented remarkable progress towards polio eradication, tackling the last 1% of polio has proved tantalizingly difficult. Pockets of endemic transmission currently persist both on the border between Afghanistan and Pakistan and in northern Nigeria. These pockets have permitted the reinfection of countries that were previously polio-free. Global strategic plans for polio eradication set out the activities, resources and financing needed to overcome the managerial, technical and security challenges faced by those tasked with the interruption of poliovirus transmission. However, polio eradication also depends on the less tangible but equally important human qualities of energy, realism, articulacy, determination, imagination, collaboration, adaptability, tactical awareness, innovation, openness and nimbleness (the initial letters of which give the acronym "ERADICATION"). By paying attention to these human qualities, the stakeholders involved may be more likely to achieve global polio eradication.


Analysis of insertion-deletion from deep-sequencing data: software evaluation for optimal detection.
Insertion and deletion (indel) mutations, the most common type of structural variance in the human genome, affect a multitude of human traits and diseases. New sequencing technologies, such as deep sequencing, allow massive throughput of sequence data and greatly contribute to the field of disease causing mutation detection, in general, and indel detection, specifically. In order to infer indel presence (indel calling), the deep-sequencing data have to undergo comprehensive computational analysis. Selecting which indel calling software to use can often skew the results and inherent tool limitations may affect downstream analysis. In order to better understand these inter-software differences, we evaluated the performance of several indel calling software for short indel (1-10 nt) detection. We compared the software's sensitivity and predictive values in the presence of varying parameters such as read depth (coverage), read length, indel size and frequency. We pinpoint several key features that assist successful experimental design and appropriate tool selection. Our study may also serve as a basis for future evaluation of additional indel calling methods.


Cardiac retransplantation in high risk pediatric patients.
Cardiac retransplantation is often the only therapy to treat GV or other causes of allograft failure. Previous reports of retransplantation have conflicting results. In this series of 18 re-transplants in 16 patients from 1984-2005, indications for retransplantation were: GV (67%); GV with cellular rejection (28%); acute graft failure (2.5%); and chronic graft failure (2.5%). Mean age at retransplantation was 12.3 (range: 0.7-22) years with a mean primary graft survival of 5.3 years (range: 8 days-10.5 years). There was no short-term mortality with only three deaths at 4, 10, and 16 years post-retransplantation. Fourteen of 18 patients had risk factors for adverse outcomes following retransplantation: ECMO support in one patient prior to retransplantation; impaired renal function in three patients; elevated panel reactive antibody screen in seven patients; a history of PTLD in five patients; and a recent episode of rejection (13-36 days) in four patients. One-, five- and ten-year survival after retransplantation was 100%, 83% and 66%, respectively, comparable to survival after primary transplantation. Freedom from rejection was not significantly different between primary and retransplantations. All patients who underwent treatment for PTLD had excellent results after retransplantation with one recurrence 16 months after retransplant. Overall, patients had excellent survival after retransplantation even in those with risk factors for poor outcome.


Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes on plastic kitchen cutting boards by electrolyzed oxidizing water.
One milliliter of culture containing a five-strain mixture of Escherichia coli O157:H7 (approximately 10(10) CFU) was inoculated on a 100-cm2 area marked on unscarred cutting boards. Following inoculation, the boards were air-dried under a laminar flow hood for 1 h, immersed in 2 liters of electrolyzed oxidizing water or sterile deionized water at 23 degrees C or 35 degrees C for 10 or 20 min; 45 degrees C for 5 or 10 min; or 55 degrees C for 5 min. After each temperature-time combination, the surviving population of the pathogen on cutting boards and in soaking water was determined. Soaking of inoculated cutting boards in electrolyzed oxidizing water reduced E. coli O157:H7 populations by > or = 5.0 log CFU/100 cm2 on cutting boards. However, immersion of cutting boards in deionized water decreased the pathogen count only by 1.0 to 1.5 log CFU/100 cm2. Treatment of cutting boards inoculated with Listeria monocytogenes in electrolyzed oxidizing water at selected temperature-time combinations (23 degrees C for 20 min, 35 degrees C for 10 min, and 45 degrees C for 10 min) substantially reduced the populations of L. monocytogenes in comparison to the counts recovered from the boards immersed in deionized water. E. coli O157:H7 and L. monocytogenes were not detected in electrolyzed oxidizing water after soaking treatment, whereas the pathogens survived in the deionized water used for soaking the cutting boards. This study revealed that immersion of kitchen cutting boards in electrolyzed oxidizing water could be used as an effective method for inactivating foodborne pathogens on smooth, plastic cutting boards.


How Blockchain can impact financial services - The overview, challenges and recommendations from expert interviewees.
FinTech (Financial Technology) and Blockchain are prevalent topics among technology leaders in finance today. This article describes the impact and revolution of FinTech and Blockchain in the financial industry and demonstrates the main characteristics of such technology. Then, we present three critical challenges as well as three ethical issues about using Blockchain technology. Next, we discuss the development of Blockchain for the financial sector. In addition, we describe the real motivations for banks to explore Blockchain, and problems they face. In order to have a good understanding of the industry, a qualitative method was adopted, and sixteen experts were interviewed. It was identified that knowledge hiding in Blockchain was common and the rationale behind was analyzed using the TPB (Theory of Planned Behavior) approach. The analysis results suggested that knowledge hiding was due to affective, behavioral and cognitive evaluations. The interviewees also provided several recommendations and success factors to overcome current issues in Blockchain adoption. Therefore, four important propositions have been developed. Finally, this article suggests how financial services should respond to this new technology and how to manage knowledge sharing in a more structured way. This article contributes to the literature related to the current entrepreneurial finance landscape for Blockchain.


Philanthropy for global mental health 2000-2015.
Mental disorders are the leading cause of years lived with disability worldwide. While over three-quarters of people with mental disorders live in low- and middle-income countries (LMICs) and effective low-cost interventions are available, resource commitments are extremely limited. This paper seeks to understand the role of philanthropy in this area and to inform discussions about how to increase investments. Novel analyses of a dataset on development assistance for health were conducted to study philanthropic development assistance for mental health (DAMH) in 156 countries between 2000 and 2015. Philanthropic contributions more than doubled over 16 years, accounting for one-third (US$364.1 million) of total DAMH 2000-2015. However, across health conditions, mental disorders received the lowest amount of philanthropic development assistance for health (0.5%). Thirty-seven of 156 LMICs received no philanthropic DAMH between 2000 and 2015 and just three LMICs (Antigua and Barbuda, Grenada, Saint Vincent and the Grenadines) received more than US$1 philanthropic DAMH per capita over the entire period. Eighty-one percent of philanthropic DAMH was disbursed to unspecified locations. Philanthropic donors are potentially playing a critical role in DAMH, and the paper identifies challenges and opportunities for increasing their impact in sustainable financing for mental health.


Lactose intolerance in infants, children, and adolescents.
The American Academy of Pediatrics Committee on Nutrition presents an updated review of lactose intolerance in infants, children, and adolescents. Differences between primary, secondary, congenital, and developmental lactase deficiency that may result in lactose intolerance are discussed. Children with suspected lactose intolerance can be assessed clinically by dietary lactose elimination or by tests including noninvasive hydrogen breath testing or invasive intestinal biopsy determination of lactase (and other disaccharidase) concentrations. Treatment consists of use of lactase-treated dairy products or oral lactase supplementation, limitation of lactose-containing foods, or dairy elimination. The American Academy of Pediatrics supports use of dairy foods as an important source of calcium for bone mineral health and of other nutrients that facilitate growth in children and adolescents. If dairy products are eliminated, other dietary sources of calcium or calcium supplements need to be provided.


Effect of peripheral circadian dysfunction on metabolic disease in response to a diabetogenic diet.
BMAL1 is a core component of the transcription/translation machinery that regulates central and peripheral circadian rhythms that coordinate behavior and metabolism, respectively. Our objective was to determine the impact of BMAL1 in adipose alone or in combination with liver on metabolic phenotypes. Control, adipose-Bmal1 knockout (ABKO), and liver- and adipose-Bmal1 knockout (LABKO) female mice were placed in TSE System metabolic chambers for metabolic phenotyping. A second cohort of male mice was fed a control or diabetogenic diet, and body weight and composition, glucose tolerance, insulin sensitivity, and serum and hepatic lipids were measured. Both female ABKO and LABKO mice exhibited increased food consumption compared with control mice. ABKO mice also exhibited increased overall activity predominantly during the light phase compared with both control and LABKO mice and were protected from increased weight gain. When the male cohort was challenged with a diabetogenic diet, LABKO mice had increased body weight due to increased fat mass compared with control and ABKO mice. However, these mice did not present further impairments in glycemic control, adipose inflammation, or liver injury. LABKO mice had increased hepatic cholesterol and elevated expression of cholesterol synthesis and uptake genes. Our data indicate that deletion of this allele in adipose or in combination with liver alters feeding behavior and locomotor activity. However, obesity is exacerbated only with the combination of liver and adipose deletion.


Website review: how to get the best from fission yeast genome data.
Researchers are increasingly depending on various centralized resources to access the vast amount of information reported in the literature and generated by systematic sequencing and functional genomics projects. Biological databases have become everyday working tools for many researchers. This dependency goes both ways in that the databases require continuous feedback from the research community to maintain accurate, reliable, and upto- date information. The fission yeast Schizosaccharomyces pombe has recently been sequenced, setting the stage for the post-genome era of this popular model organism. Here, we provide an overview of relevant databases available, or being developed, together with a compilation of Internet resources containing useful information and tools for fission yeast.


GroEL accelerates the refolding of hen lysozyme without changing its folding mechanism.
The chaperonin GroEL binds folding intermediates of four-disulfidehen lysozyme transiently within its central cavity. Using stopped flow fluorescence we show that GroEL binds early intermediates in folding and accelerates the slow kinetic phase that reflects the reversal of non-native interactions involving tryptophan residues and the formation of the native state. Pulsed hydrogen exchange monitored by electrospray ionization mass spectrometry demonstrates that GroEL does not alter the folding mechanism, nor are protected species unfolded by the chaperonin. The data suggest a mechanism for GroEL-assisted folding in which the reorganization of non-native tertiary interactions is facilitated but domain folding is unperturbed.


Persistent brain network homology from the perspective of dendrogram.
The brain network is usually constructed by estimating the connectivity matrix and thresholding it at an arbitrary level. The problem with this standard method is that we do not have any generally accepted criteria for determining a proper threshold. Thus, we propose a novel multiscale framework that models all brain networks generated over every possible threshold. Our approach is based on persistent homology and its various representations such as the Rips filtration, barcodes, and dendrograms. This new persistent homological framework enables us to quantify various persistent topological features at different scales in a coherent manner. The barcode is used to quantify and visualize the evolutionary changes of topological features such as the Betti numbers over different scales. By incorporating additional geometric information to the barcode, we obtain a single linkage dendrogram that shows the overall evolution of the network. The difference between the two networks is then measured by the Gromov-Hausdorff distance over the dendrograms. As an illustration, we modeled and differentiated the FDG-PET based functional brain networks of 24 attention-deficit hyperactivity disorder children, 26 autism spectrum disorder children, and 11 pediatric control subjects.


[Epidemiological survey on paragonimiasis in Kang Hwa Gun]
This study was undertaken to evaluate the status of human paragonimiasis in Kang Hwa Gun, Kyunggi Do, Korea from March 1973 to October 1975 by the use of intradermal screening test with Veronal buffered saline antigen. A total number of 7,352 (male 3,936; female 3,4l6) consisting of 3,447 primary school children, 2,689 middle school students, l,008 high school students of 9 Myuns out of 13 Myuns of this County and 208 residents of all age group in Kilgeong-Ri, Yangdo Myun were examined by the intradermal test with Paragonimus antigen. In order to detect Paragonimus egg, the sputum and the stool examinations were undertaken on the residents in Kilgeong-Ri. The infestation rate of crayfish with Paragonimus metacercaria was also examined in Konneung and Jeongha village in Kilgeong-Ri. The following results were obtained in this survey: 1. As a whole, 16.6 % of the examiners showed positive reaction with sexual difference of 16.1% in male and 14.8 % in female. 2. In age groups, the highest positive rate showed in middle school age group (17.1 %) than primary school (14.0 %) and high school age group (11.4 %). 3. In regional differences of positive rates, the highest rate (28.4 %) showed at Yangdo Myun followed by Kilsang Myun (27.0 %), Hwado Myun (25.0 %), and Naega Myun(20.8 %). 4. In the result of 208 residents Kilgeong-Ri, the positive of 36.1 % in male and 20.2 % in female. The highest positive rate showed in 10 to 19 age group (41.5 %). 5. Among the residents in Kilgeong-Ri, Paragonimus eggs were detected in 11 residents by the sputum and the stool examinations. 6. The infestation rate of crayfsh with Paragonimus metacercariae was 100 % (41 out of 41 crayfishes) in Konneung village, and 84.6 % (9 out of 13 crayfishes) in Jeongha village, Kilgeong-Ri. 7. The fluctuation of the infection rate of crayfish with Paragonimus metacercariae was decreased in Konneung village in 2 years after the treatment with Bilevon against 10 paragonimiasis patients.


Signal-transduction pathways causing slow synaptic excitation in guinea pig myenteric AH neurons.
Intracellular recordings were obtained from myenteric AH neurons of guinea pig ileum in vitro. Slow excitatory synaptic responses associated with decreased potassium conductance (gK), inhibition of the spike afterhyperpolarization current (AHC), and increased chloride conductance (gCl) were mimicked by senktide, a neurokinin3 receptor agonist. Intracellular guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) decreased gK and increased gCl irreversibly after nerve stimulation or senktide application. Myenteric neurons in pertussis toxin (PTX)-treated tissues responded normally to senktide and nerve stimulation. Forskolin and phorbol 12,13-dibutyrate (PDBu) inhibited gK and the AHC but did not activate gCl. The AHC was not reduced by subthreshold concentrations of forskolin (10 nM) or PDBu (3 nM) alone but was inhibited by forskolin and PDBu applied together. Inhibitors of phospholipase C (D-609) or protein kinases (staurosporine) reduced slow synaptic and senktide responses. The protein phosphatase inhibitor, calyculin A, caused an inward current, a decrease in gK, and AHC inhibition but did not activate gCl. We conclude that slow excitatory synaptic responses are mediated by PTX-insensitive G proteins and activation of phospholipase C and protein kinases. Forskolin and PDBu activate pathways that inhibit gK. The mechanisms for activation of gCl are unknown.


How we can change your mind: Anodal tDCS to Fp3 alters human stimulus representation and learning.
The aim of the current work is to advance our understanding of both the mechanisms controlling perceptual learning and the face inversion effect. In the three double blind experiments reported here (total N = 144) we have shown that anodal tDCS stimulation (10 min at 1.5 mA) delivered over the left DLPFC at Fp3 affects perceptual learning and drastically reduces the, usually robust, face inversion effect. In Experiment 1, we found a significantly reduced inversion effect in the anodal group compared to that in the sham group. Experiment 2 replicated the pattern of results found in Experiment 1. In both experiments recognition performance for upright faces in the anodal group was significantly impaired compared to that in the sham group. Finally, using an active control in Experiment 3 (same behavioural task but different tDCS targeted brain area) we showed that the same Fp3 anodal tDCS stimulation effect is not obtained when a different brain area is targeted.


Sleep-dependent consolidation in children with comprehension and vocabulary weaknesses: it'll be alright on the night?
Vocabulary is crucial for an array of life outcomes and is frequently impaired in developmental disorders. Notably, 'poor comprehenders' (children with reading comprehension deficits but intact word reading) often have vocabulary deficits, but underlying mechanisms remain unclear. Prior research suggests intact encoding but difficulties consolidating new word knowledge. We test the hypothesis that poor comprehenders' sleep-associated vocabulary consolidation is compromised by their impoverished lexical-semantic knowledge. Memory for new words was tracked across wake and sleep to assess encoding and consolidation in 8-to-12-year-old good and poor comprehenders. Each child participated in two sets of sessions, one beginning in the morning (AM-encoding) and the other in the evening (PM-encoding). In each case, they were taught 12 words and were trained on a spatial memory task. Memory was assessed immediately, 12- and 24-hr later via stem-completion, picture-naming, and definition tasks to probe different aspects of word knowledge. Long-term retention was assessed 1-2 months later. Recall of word-forms improved over sleep and postsleep wake, as measured in both stem-completion and picture-naming tasks. Counter to hypotheses, deficits for poor comprehenders were not observed in consolidation but instead were seen across measures and throughout testing, suggesting a deficit from encoding. Variability in vocabulary knowledge across the whole sample predicted sleep-associated consolidation, but only when words were learned early in the day and not when sleep followed soon after learning. Poor comprehenders showed weaker memory for new words than good comprehenders, but sleep-associated consolidation benefits were comparable between groups. Sleeping soon after learning had long-lasting benefits for memory and may be especially beneficial for children with weaker vocabulary. These results provide new insights into the breadth of poor comprehenders' vocabulary weaknesses, and ways in which learning might be better timed to remediate vocabulary difficulties.


Quercetin ameliorates paclitaxel-induced neuropathic pain by stabilizing mast cells, and subsequently blocking PKCε-dependent activation of TRPV1.
Severe painful sensory neuropathy often occurs during paclitaxel chemotherapy. Since paclitaxel can activate mast cell and basophils, whereas quercetin, a polyphenolic flavonoid contained in various plants, which can specifically inhibit histamine release as a mast cell stabilizer. In this study we explore whether quercetin could ameliorate paclitaxel-induced neuropathic pain and elucidated the underlying mechanisms. Quercetin inhibition on histamine release was validated in vitro by detecting histamine release from rat basophilic leukemia (RBL-2H3) cells stimulated with paclitaxel (10 μmol/L). In the in vivo experiments, rats and mice received quercetin (20, 40 mg·kg(-1)·d(-1)) for 40 and 12 d, respectively. Meanwhile, the animals were injected with paclitaxel (2 mg/kg, ip) four times on d 1, 3, 5 and 7. Heat hyperalgesia and mechanical allodynia were evaluated at the different time points. The animals were euthanized and spinal cords and dorsal root ganglions were harvested for analyzing PKCε and TRPV1 expression levels. The plasma histamine levels were assessed in rats on d 31. Pretreatment with quercetin (3, 10, 30 μmol/L) dose-dependently inhibited excessive histamine release from paclitaxel-stimulated RBL-2H3 cells in vitro, and quercetin administration significantly suppressed the high plasma histamine levels in paclitaxel-treated rats. Quercetin administration dose-dependently raised the thresholds for heat hyperalgesia and mechanical allodynia in paclitaxel-treated rats and mice. Furthermore, quercetin administration dose-dependently suppressed the increased expression levels of PKCε and TRPV1 in the spinal cords and DRGs of paclitaxel-treated rats and mice. Moreover, quercetin administration may inhibited the translocation of PKCε from the cytoplasm to the membrane in the spinal cord and DRG of paclitaxel-treated rats. Our results reveal the underlying mechanisms of paclitaxel-induced peripheral neuropathy and demonstrate the therapeutic potential of quercetin for treating this side effect.


Luteolin, a flavone, does not suppress postprandial glucose absorption through an inhibition of alpha-glucosidase action.
In order to clarify the postprandial glucose suppression via alpha-glucosidase (AGH) inhibitory action by natural compounds, flavonoids were examined in this study. Among the flavonoids (luteolin, kaempferol, chrysin, and galangin), luteolin showed the potent maltase inhibitory activity with the IC50 of 2.3 mM, while less inhibitions were observed against sucrase. In addition, the effects of maltase inhibition by flavonoids were observed in the descending order of potency of luteolin > kaempferol > chrysin > galangin. Apparently, the AGH inhibition power greatly increased with the replacement of hydroxyl groups at 3' and 4'-position of the B-ring. However, the inhibitory power of luteolin was poorer than a therapeutic drug (acarbose: IC50; 430 nM). As a result of a single oral administration of maltose or sucrose (2 g/kg) in SD rats, no significant change in blood glucose level with the doses of 100 and 200 mg/kg of luteolin was observed. These findings strongly suggested that luteolin given at less than 200 mg/kg did not possess the ability to suppress the glucose production from carbohydrates through the inhibition of AGH action in the gut.


Acute myocardial infarction: profile and management at a tertiary care hospital in Karachi.
Acute Myocardial Infarction (AMI) is a rising epidemic in developing countries. While studies in the West have established the characteristics and management of AMI patients, comprehensive data reflecting these issues in the Pakistani subjects is scarce. This study examined the profile and management of AMI in patients hospitalized at a tertiary care hospital in Karachi, Pakistan. Three hundred forty four patients admitted in 1998 with the diagnosis of AMI met our inclusion criteria. Data on presentation, investigations, monitoring and therapy was obtained. Chi-square and t tests were used to analyze the data. Out of 344 patients with AMI, 71% were males; 58% had a Q wave MI. Majority of the patients who presented within 2 hours of symptom onset (36%), had chest pain. Patients with dyspnea and no chest pain were more likely to present after 12 hours of the onset of symptoms. In-house mortality was found to be 10.8%. Low HDL and diabetes was associated with in-hospital complications. Twenty nine percent of patients were given thrombolytic therapy with a mean door-to-needle time of 1 hour 36 minutes; 33% of patients who were eligible of Streptokinase did not receive it. Cardiac catheterization was performed in 28% patients. Echocardiography and Exercise Tolerance Test, both under utilized, were performed in 67% and 16% of patients, respectively. Two hundred sixteen (70%) patients discharged from hospital were contacted via telephone and the 1-year mortality rate among them was 28%. The profile and management of AMI was in coherence with earlier, Western studies. Chest pain units need to be established in the Emergency Room. Patients should be risk stratified prior to discharge. Public awareness regarding primary and secondary prevention and symptoms of AMI needs to be increased.


Systemic shigellosis in South Africa.
Systemic disease due to shigellae is associated with human immunodeficiency virus (HIV), malnutrition, and other immunosuppressed states. We examined the clinical and microbiologic characteristics of systemic shigellosis in South Africa, where rates of HIV infection are high. From 2003 to 2009, 429 cases of invasive shigellosis were identified through national laboratory-based surveillance. At selected sites, additional information was captured on HIV serostatus and outcome. Isolates were serotyped and antimicrobial susceptibility testing performed. Most cases of systemic shigellosis were diagnosed on blood culture (408 of 429 cases; 95%). HIV prevalence was 67% (80 of 120 cases), highest in patients aged 5-54 years, and higher among females (55 of 70 cases; 79%) compared with males (25 of 48 cases; 52%; P = .002). HIV-infected people were 4.1 times more likely to die than HIV-uninfected cases (case-fatality ratio, 29 of 78 HIV-infected people [37%] vs 5 of 40 HIV-uninfected people [13%]; P = .008; 95% confidence interval [CI], 1.5-11.8). The commonest serotype was Shigella flexneri 2a (89 of 292 serotypes [30.5%]). Pentavalent resistance occurred in 120 of 292 isolates (41.1%). There was no difference in multidrug resistance between HIV-infected patients (33 of 71 [46%]) and uninfected patients (12 of 33 [36%]; 95% CI, .65--3.55). Systemic shigellosis is associated with HIV-infected patients, primarily in older girls and women, potentially due to the burden of caring for sick children in the home; interventions need to be targeted here. Death rates are higher in HIV-infected versus uninfected individuals.


Cardiac Rehabilitation Utilization During an Acute Cardiac Hospitalization: A NATIONAL SAMPLE.
Inpatient cardiac rehabilitation (ICR) programs provide important services to hospitalized patients by delivering risk factor education, daily ambulation, and facilitation of referral to outpatient cardiac rehabilitation. However, little is known about ICR utilization or practice patterns. We examined the use of ICR, between January 2007 and June 2011, in a geographically and structurally diverse sample of US hospitals (Premier, Inc). Among 458 hospitals, there were 1 343 537 admissions with a qualifying diagnosis for outpatient cardiac rehabilitation. Formal ICR was available at 223 (49%) of these hospitals. Overall, patient utilization of ICR was low (21.2%) and varied by indication. Utilization was highest in those undergoing cardiac surgery (43.3%) and lowest in patients with medically managed myocardial infarction (15.6%) or heart failure (10.6%). A larger bed count, the presence of cardiac interventional services, and Midwest location were associated with increased likelihood of a hospital having an ICR program. In multivariable hierarchical analysis adjusting for known hospital characteristics among hospitals that provided ICR, multiple patient factors were associated with a lower likelihood of ICR utilization, including older age, more comorbidities, female sex, and Medicare insurance, but unspecified hospital characteristics explained the vast majority of the variability. We found substantial variation in the delivery of ICR across US hospitals and by patient condition. Overall, only a minority of eligible patients ever received ICR and fewer than half of hospitals treating cardiac patients provided formal ICR services. This substantial gap in the secondary prevention of heart disease warrants further investigation and intervention.


It's time for change; commentary in response to feature article by Halpin et al.
It is essential that we re-examine the way new drugs for life-threatening illnesses are assessed and regulated. The time taken to approve lifesaving medicines for general use outside of clinical trials is prohibitively slow; people are needlessly dying. The bioethics community should be at the forefront of change. In particular, the rigid adherence to Evidence Based Medicine (EBM) has become irrational. New regulatory frameworks are being proposed, including that of adaptive licensing. Whereas the current model uses a one-point-in-time, all-or-nothing system of regulation, adaptive licensing allows for progressive access to a new drug during the approval process. Indeed this may be the first instance where the contemporary practice of EBM needs to be abandoned due to inherent inefficiencies. In response to the inefficiencies of the current system, a collaboration of stakeholders has established NEWDIGS (NEW Drug Development ParadIGmS). The Feature Article by Halpin and colleagues, on which this Commentary is based, gives a sophisticated survey of relevant bioethical issues. In addition this Commentary outlines a novel bioethical model to align with proposed new regulatory frameworks: a distributive-benefit-at-equilibrium model.


Process Evaluation Results From an Intergenerational Physical Activity Intervention for Grandparents Raising Grandchildren.
Grandparents and the grandchildren they raise may experience stress related to their caregiving relationship that negatively impacts their health. Thus, there is a need to develop intergenerational health promotion interventions for these kinship families. An 8-week intergenerational physical activity intervention for kinship families was developed and implemented. The specific goal was to understand the process of implementing the intervention. Content analysis of observational data provided an in-depth account of the intervention's process (ie, recruitment, dose delivered, dose received, fidelity, and context). Community and support service organizations referred more participants to the study than individual stakeholders. Most participants attended approximately 10 classes, and the grandparents were more engaged than the grandchildren during the classes. Intervention fidelity was confirmed with the fidelity checklist and observational notes. Health emerged as a barrier to participation, while the intergenerational nature of the intervention was a facilitator. Lastly, the context domain described how the grandparents' complex lives affected their ability to participate, while the dedication of the recreation staff helped the intervention to proceed efficiently. The distinct details gleaned from this study can provide guidance on how to develop and implement future intergenerational interventions.


Chiton integument: development of sensory organs in juvenile Mopalia muscosa.
The girdle epidermis of adult Mopalia muscosa secretes several types of structures, including calcareous spicules and innervated hairs. Newly metamorphosed chitons superficially resemble adult animals, but they lack the adult girdle ornaments, shell sculpture, and coloration. The morphogenesis of the adult girdle structures has not been described previously for any species. Juvenile Mopalia muscosa secrete hairs at metamorphosis, but it was not known if these hairs were sensory or if they were retained as the animals grew. I discovered that the hairs of juveniles become the tips of adult hairs. When juvenile hairs are detectable by light microscopy the sensory components already exist, suggesting that they are functional receptor organs. The other girdle ornaments of young juveniles, the primary calcareous spicules, are lost as the animal grows. I also demonstrated that the hairs are not uniquely innervated; the same sensory structures are produced in conjunction with other girdle ornaments on the marginal and ventral faces.


Genotyping markers used for multi locus VNTR analysis with ompA (MLVA-ompA) and multi sequence typing (MST) retain stability in Chlamydia trachomatis.
We aimed to evaluate the stability of the Chlamydia trachomatis multi locus VNTR analysis (MLVA-ompA) and multi sequence typing (MST) systems through multiple passages in tissue culture. Firstly, we analyzed the stability of these markers through adaptation of C. trachomatis to tissue culture and secondly, we examined the stability of a four-locus MLVA-ompA and a five-locus MST system after multiple passages in tissue culture. Marker sequences were monitored through successive chlamydial developmental cycles to evaluate the stability of the individual DNA markers through many bacterial divisions and this, in turn, informed us of the usefulness of using such typing systems for short and long-term molecular epidemiology. Southampton genitourinary medicine (GUM) clinic isolates from endocervical swabs collected from C. trachomatis positive women were passaged through tissue culture. MLVA-ompA typing was applied to primary swab samples and to the same samples after C. trachomatis had been passaged through cell culture (eight passages). Sequence data from time-zero and passage-eight isolates were aligned with reference sequences to determine the stability of the markers. The Swedish new variant (nvCT) underwent 72 passages in cell culture and the markers of the two schemes were similarly analyzed. Analysis of genetic markers of the MLVA-ompA typing system before and after the isolates were introduced to tissue culture showed no change in the dominant sequence. The nvCT that had been passaged 72 times over the duration of a year also showed no variation in the dominant sequence for both the genotyping schemes. MLVA-ompA and MST markers are stable upon adaptation of C. trachomatis to tissue culture following isolation of strains from primary endocervical swab samples. These markers remain stable throughout multiple rounds of cell-division in tissue culture, concomitant with the incubation period and appearance of symptoms normally associated with host-infection. Both genotyping schemes are, therefore, suitable for epidemiology of C. trachomatis.


Herpes simplex virus type 1 ICP0 plays a critical role in the de novo synthesis of infectious virus following transfection of viral DNA.
As a first step in identifying the functions and intramolecular functional domains of herpes simplex virus type 1 infected cell protein 0 (ICP0) in productive infection and latency, a series of mutant plasmids specifying varying amounts of the ICP0 primary amino acid sequence were constructed. In transient expression assays with mutant and wild-type plasmids, the N-terminal half of the ICP0 molecule was found to be sufficient to transactivate a variety of viral promoters. Although promoters representing the immediate-early, early, and late kinetic classes were transactivated by wild-type ICP0, individual promoters responded to mutant forms of ICP0 in a manner consistent with the possibility that ICP0 transactivates different promoters by different mechanisms. Unlike infection with virus particles, which contain the 65-kilodalton transcriptional transactiovator, the initiation of viral replication after transfection of cells with purified viral DNA requires de novo protein synthesis. In order to assess the role of ICP0 in the de novo synthesis of infectious virus, Vero cells were transfected with purified DNA of wild-type virus or an ICP0 null mutant and the production of infectious virus was monitored. In cells transfected with mutant DNA, virus production was delayed by 2 days and the level of virus was reduced by several orders of magnitude relative to Vero cells transfected with wild-type viral DNA, suggesting an important role for ICP0 in the de novo synthesis of infectious particles. In cotransfection experiments with infectious DNA of the ICP0 null mutant and a plasmid specifying wild-type ICP0 titers of infectious virus were significantly enhanced relative to transfection with mutant DNA alone, confirming the role of ICP0 in de novo synthesis. These findings are consistent with the proposed role of ICP0 in reactivation of herpes simplex virus from latency (D. A. Leib, D. M. Coen, C. L. Bogard, K. A. Hicks, D. R. Yager, D. M. Knipe, K. L. Tyler, and P. A. Schaffer, J. Virol. 63:759-768, 1989), a process also thought to require de novo protein synthesis. The complementing activities of ICP0 mutant plasmids for ICP0 null mutant DNA in cotransfection assays correlated well with their transactivating activities for viral promoters in transient assays, indicating that the transactivating function of ICP0 is a critical factor in the de novo synthesis of infectious particles.(ABSTRACT TRUNCATED AT 400 WORDS)


High-level expression of the ER-MP58 antigen on mouse bone marrow hematopoietic progenitor cells marks commitment to the myeloid lineage.
Studies on the early events in the differentiation of the nonspecific immune system require the identification and isolation of myeloid-committed progenitor cells. Using the monoclonal antibodies (mAb) ER-MP12 and ER-MP20, generated against immortalized macrophage precursors, we have shown previously that the earliest macrophage colony-stimulating factor (M-CSF)-responsive cells in the bone marrow have the ER-MP12hi 20- phenotype. In addition, we found that the ER-MP12hi 20- subset (comprising about 2 % of total nucleated marrow) contains progenitor cells of all hematopoietic lineages. Aiming at the identification and purification of the myeloid progenitor cells within the ER-MP12hi 20-subset, we used ER-MP58, a marker expressed at high level by all M-CSF-responsive bone marrow progenitors. With this marker the ER-MP12hi 20- cell population could be divided into three subfractions: one with absent or low level ER-MP58 expression, one with intermediate, and one with high ER-MP58 expression. These subfractions were isolated by fluorescence-activated cell sorting and tested in vitro and in vivo for their differentiation capacities. In addition, the expression of ER-MP58 on stem cell subsets was examined in the cobblestone area-forming cell (CAFC) assay. Our data indicate that in the ER-MP12hi 20- subpopulation myeloid-committed progenitors are characterized by high-level expression of the ER-MP58 antigen, whereas cells with other or broader differentiation capacities have an ER-MP58 negative/low or intermediate phenotype. These myeloid-committed progenitors have no significant repopulating ability in vivo, in contrast to the ER-MP58 intermediate cells. Primitive CAFC-28/35, corresponding to cells providing long-term hematopoietic engraftment in vivo, also did not express the ER-MP58 Ag at a high level. Thus, cells committed to the myeloid lineage can be separated from progenitor cells with other differentiation capacities by means of multiparameter cell sorting using ER-MP58 in combination with ER-MP12 and ER-MP20.


Colloidal nanocrystals as LEGO® bricks for building electronic band structure models.
The synthesis of self-assembled semiconductor nanocrystal (NC) superlattices using oriented attachment recently became a flourishing research topic. This technique already produced remarkable forms of NC superlattices, such as linear chains, mono and multilayer square lattices, and silicene-like honeycomb lattices. In the case of lead chalcogenide semiconductors where NCs are in the form of truncated nanocubes, the attachment mostly occurs via (100) facets. In this work, we show that all these structures can be seen as sub-structures of a simple cubic lattice. From this, we investigate a rich variety of one-dimensional or two-dimensional superlattices that could be built as few lines or few layers taken from the same cubic system following different crystallographic orientations. Each NC can be therefore considered as a LEGO® brick, and any superlattice can be obtained from another one by rearranging the bricks. Moreover, we show that this concept of LEGO® bricks can be extended to the calculation of the electronic band structure of the superlattices. This leads to a simple yet powerful way to build analytical Hamiltonians that present band structures in excellent agreement with more elaborate atomistic tight-binding calculations. This LEGO® concept could guide the synthesis of superlattices and LEGO® Hamiltonians should greatly simplify further studies on the (opto-)electronic properties of such structures.


Very recent onset arthritis--clinical, laboratory, and radiological findings during the first year of disease.
To describe clinical and radiological findings in patients with very early arthritis (< 3 months of symptoms) during one year of observation. In an Austrian multicenter setting, patients were eligible if they had nontraumatic swelling or pain in at least one joint and laboratory signs of inflammation [elevated erythrocyte sedimentation rate, C-reactive protein, leukocytosis, or rheumatoid factor (RF)] within the last 3 months. Clinical and laboratory assessments were performed every 3 months. Radiographs of hands and feet were taken at entry and after one year. Treatment decisions were left to the discretion of the participating center. In total, 108 patients included between 1996 and 2000 had followup investigations during at least one year; 61.1% of these patients had rheumatoid arthritis (RA). Over 65% of RA diagnoses were made at the first visit. Lag time to referral was significantly longer in patients with RA than in patients with other inflammatory joint diseases (median 8 vs 4 weeks). Disease modifying antirheumatic drugs were started 19 +/- 10 (mean +/- SD) weeks after symptom onset in patients with RA. Patients with RA improved significantly (by American College of Rheumatology response criteria and the Disease Activity Score 28) during the first year. Erosions were present in 12.8% of RA patients' initial radiographs, compared to 27.6% after one year. Odds ratio to develop new erosions during the first year of RA was 9.7 (95% CI 1.05-89.93) in RF+ patients compared to RF- individuals (p < 0.05). When early referral of patients with arthritis is encouraged, RA can be diagnosed and treatment initiated early, with significant clinical response. Moreover, patients with RA tend to be referred later than patients with other inflammatory joint diseases; RA patients at this very early stage have low frequency of joint damage; and RF predicts erosions in the first year.


Myogenic satellite cells: physiology to molecular biology.
Adult skeletal muscle has a remarkable ability to regenerate following myotrauma. Because adult myofibers are terminally differentiated, the regeneration of skeletal muscle is largely dependent on a small population of resident cells termed satellite cells. Although this population of cells was identified 40 years ago, little is known regarding the molecular phenotype or regulation of the satellite cell. The use of cell culture techniques and transgenic animal models has improved our understanding of this unique cell population; however, the capacity and potential of these cells remain ill-defined. This review will highlight the origin and unique markers of the satellite cell population, the regulation by growth factors, and the response to physiological and pathological stimuli. We conclude by highlighting the potential therapeutic uses of satellite cells and identifying future research goals for the study of satellite cell biology.


Gene-environment interactions modulating cognitive function and molecular correlates of synaptic plasticity in Huntington's disease transgenic mice.
Huntington's disease (HD) is a fatal neurodegenerative disorder characterized by motor, cognitive and psychiatric symptoms. Here, we show that R6/1 (HD) mice have deficits in short-term hippocampal-dependent memory prior to onset of motor symptoms. HD mice also exhibit impaired performance on a test of long-term spatial memory, however, environmental enrichment enhanced spatial learning and significantly ameliorated this memory deficit in HD mice. Analysis of the presynaptic vesicle protein synaptophysin showed no differences between standard-housed wild-type and HD littermates, however, enrichment increased synaptophysin levels in the frontal cortex and hippocampus in both groups. In comparison, analysis of postsynaptic proteins revealed that HD animals show decreased levels of PSD-95 and GluR1, but no change in levels of gephyrin. Furthermore, at 12 weeks of age when we observe a beneficial effect of enrichment on spatial learning in HD mice, enrichment also delays the onset of a deficit in hippocampal PSD-95 levels. Our results show that cognitive deficits in HD mice can be ameliorated by environmental enrichment and suggest that changes in synaptic composition may contribute to the cognitive alterations observed.


σB Inhibits Poly-N-Acetylglucosamine Exopolysaccharide Synthesis and Biofilm Formation in Staphylococcus aureus.
Staphylococcus aureus clinical strains are able to produce at least two distinct types of biofilm matrixes: biofilm matrixes made of the polysaccharide intercellular adhesin (PIA) or poly-N-acetylglucosamine (PNAG), whose synthesis is mediated by the icaADBC locus, and biofilm matrixes built of proteins (polysaccharide independent). σB is a conserved alternative sigma factor that regulates the expression of more than 100 genes in response to changes in environmental conditions. While numerous studies agree that σB is required for polysaccharide-independent biofilms, controversy persists over the role of σB in the regulation of PIA/PNAG-dependent biofilm development. Here, we show that genetically unrelated S. aureus σB-deficient strains produced stronger biofilms under both static and flow conditions and accumulated higher levels of PIA/PNAG exopolysaccharide than their corresponding wild-type strains. The increased accumulation of PIA/PNAG in the σB mutants correlated with a greater accumulation of the IcaC protein showed that it was not due to adjustments in icaADBC operon transcription and/or icaADBC mRNA stability. Overall, our results reveal that in the presence of active σB, the turnover of Ica proteins is accelerated, reducing the synthesis of PIA/PNAG exopolysaccharide and consequently the PIA/PNAG-dependent biofilm formation capacity.IMPORTANCE Due to its multifaceted lifestyle, Staphylococcus aureus needs a complex regulatory network to connect environmental signals with cellular physiology. One particular transcription factor, named σB (SigB), is involved in the general stress response and the expression of virulence factors. For many years, great confusion has existed about the role of σB in the regulation of the biofilm lifestyle in S. aureus Our study demonstrated that σB is not necessary for exopolysaccharide-dependent biofilms and, even more, that S. aureus produces stronger biofilms in the absence of σB The increased accumulation of exopolysaccharide correlates with higher stability of the proteins responsible for its synthesis. The present findings reveal an additional regulatory layer to control biofilm exopolysaccharide synthesis under stress conditions.


Identification of an alpha-tubulin mutant of fission yeast from gamma-tubulin-interacting protein screening: genetic evidence for alpha-/gamma-tubulin interaction.
gamma-Tubulin has been determined to be a central element of microtubule nucleation and, thus, indispensable for cellular organization of the microtubule. Utilizing the fact that human gamma-tubulin can function in the fission yeast Schizosaccharomyces pombe, we have generated a unique mutant screening procedure which can specifically select mutants of genes encoding gamma-tubulin-interacting proteins. One of the isolated mutants, cs76, turned out to carry a mutation in the alpha 1-tubulin gene (nda2(+)). This result suggests a direct interaction between the alpha- and gamma-tubulins. We located the mutation site in the nda2 gene and characterized the mutant phenotype. Our results demonstrate the importance of the alpha-/gamma-tubulin interaction in microtubule nucleation and should complement previous knowledge.


Conserved regions in the Epstein-Barr virus leader protein define distinct domains required for nuclear localization and transcriptional cooperation with EBNA2.
Epstein-Barr virus (EBV) EBNA-LP is a latent protein whose function is not fully understood. Recent studies have shown that EBNA-LP may be an important EBNA2 cofactor by enhancing EBNA2 stimulation of the latency C and LMP-1 promoters. To further our understanding of EBNA-LP function, we have introduced a series of mutations into evolutionarily conserved regions and tested the mutant proteins for the ability to enhance EBNA2 stimulation of the latency C and LMP-1 promoters. Three conserved regions (CR1 to CR3) are located in the repeat domains that are essential for the EBNA2 cooperativity function. In addition, three serine residues are also well conserved in the repeat domains. Clustered alanine mutations were introduced into CR1 to CR3, and the conserved serines were also changed to alanine residues in an EBNA-LP with two repeats, which is the minimal protein able to cooperate with EBNA2. Mutations introduced into CR1a had no effect on EBNA-LP function, while mutations introduced into CR1b resulted in EBNA-LP with slightly decreased activity. Mutations in CR1c and CR2 resulted in proteins that no longer localized exclusively to the nucleus and also had no EBNA2 cooperation activity. Mutations introduced into conserved serines S5/71 resulted in proteins with slightly higher activity, while mutations introduced into conserved serines S35/101 or in CR3 (which contains S60/126) resulted in EBNA-LP proteins with substantially reduced activity. The potential karyophilic signals within EBNA-LP CR1c and CR2 were also examined by introducing oligonucleotides encoding these positively charged amino acid groupings into a cytoplasmic test protein, herpes simplex virus DeltaIE175, and by examining the intracellular localization of the resulting proteins. This assay identified a strong nuclear localization signal between EBNA-LP amino acids 43 and 50 (109 to 117 in the second W repeat) comprising CR2, while EBNA-LP amino acids 29 to 36 (91 to 98 in the second W repeat) were unable to function independently as a nuclear localization signal. However, a combination of amino acids 29 to 50 resulted in more efficient nuclear localization than with amino acids 43 to 50 alone. These results indicate that EBNA-LP has a bipartite nuclear localization signal and that efficient nuclear localization is essential for EBNA2 cooperativity function. Interestingly, EBNA-LP with only a single repeat localized exclusively to the cytoplasm, providing an explanation for why this isoform has no activity. In addition, two conserved serine residues which are distinct from nuclear import functions are important for EBNA2 cooperativity function.


Deletion of the TSC2 and PKD1 genes associated with severe infantile polycystic kidney disease--a contiguous gene syndrome.
Major genes which cause tuberous sclerosis (TSC) and autosomal dominant polycystic kidney disease (ADPKD), known as TSC2 and PKD1 respectively, lie immediately adjacent to each other on chromosome 16p. Renal cysts are often found in TSC, but a specific renal phenotype, distinguished by the severity and infantile presentation of the cystic changes, is seen in a small proportion of cases. We have identified large deletions disrupting TSC2 and PKD1 in each of six such cases studied. Analysis of the deletions indicates that they inactivate PKD1, in contrast to the mutations reported in ADPKD patients, where in each case abnormal transcripts have been detected.


Assessing Related Factors of Intention to Perpetrate Dating Violence among University Students Using the Theory of Planned Behavior.
Dating violence (DV) is a major public health problem among youth. The majority of DV studies in Taiwan involve the assessment of DV without the use of a robust psychological framework to guide research accuracy. Therefore, the purpose of this study was to utilize the Theory of Planned Behavior (TPB) to assess intention and other salient factors related to DV among university students. A two-phase, mixed-method design study was conducted among university students from seven universities in Taiwan, aged 18 to 24 years. In Phase I, questionnaires used were specifically developed based on the TPB, consisting of both direct and indirect measures related to DV. In Phase Ⅱ, questionnaire evaluation and related factors were examined through a two-step process of structural equation modelling (SEM) to test the TPB model. The results of this study found that perceived behavioral control, subjective norm, and attitude toward DV on behavioral intention accounting for 37.5% of the total variance. Among the related factors, attitude toward the behavior was the strongest (β = 0.48, p < 0.001), followed by perceived behavioral control (β = 0.19, p < 0.05). Findings from this study could expand the knowledge base in this important area and might help prevent DV.


Tolerance to IDDM induced by CD4 antibodies in nonobese diabetic mice is reversed by cyclophosphamide.
IDDM can be induced in nonobese diabetic (NOD) mice in several ways, including high doses of cyclophosphamide and transfer of diabetic spleen cells to sublethally irradiated recipients. It has previously been established that transferred diabetes can be prevented by treatment with a nondepleting CD4 monoclonal antibody; however, we report herein that cyclophosphamide-induced diabetes also can be prevented using this antibody. The protection induced by CD4 monoclonal antibody to transferred diabetes is maintained for a long period after cessation of antibody treatment. However, cyclophosphamide can abrogate this induced tolerance and we report that this abrogation does not require new T-cells. During the course of the experimental work described, we observed that the thymus had a suppressive effect on the expression of transferred disease. Mice that were depleted of their peripheral T-cells showed a doubling of the time for disease expression if they were euthymic, compared with thymectomized mice.