Mapping of the mouse atrial natriuretic factor gene. Evidence for tight linkage to the Fv-1 locus.
The importance of atrial natriuretic factor (ANF) in the regulation of salt balance and blood pressure is now widely recognized, and it would be extremely informative for both physiologists and molecular biologists to have mutants available that exhibit abnormal expression of this peptide hormone. The most direct mutation would affect the structural gene itself by altering either its coding potential or its regulation. With this aim in mind, it is of considerable interest to accurately determine the chromosomal position of the ANF gene (Anf) in the mouse. This information would permit a comparison with known mutations and provide a means of screening for mutations at this locus. Using recombinant inbred mouse strains, we undertook the mapping of the mouse Anf gene and demonstrated linkage of the gene to the Friend virus susceptibility-1 (Fv-1) locus on chromosome 4. The mapping was performed using a restriction fragment length polymorphism extant between the two parental strains. No recombination event between Anf and the Fv-1 locus was evident in any of the 34 strains tested. The assignment of Anf to this region of chromosome 4 coincides with the reported position of the cribriform degeneration mutation, which includes in its phenotype an abnormal electrolyte distribution. We have, therefore, begun studying this mutation to determine whether a defect in ANF expression is the underlying cause of the phenotype. Thus far, we have found no major DNA rearrangement close to the Anf gene.


All-optical modulation and switching by a metamaterial of plasmonic circuits.
We demonstrate experimentally the modulation and switching of one light beam by a second beam using metamaterials constructed from arrays of plasmonic circuits. Each circuit consists of three gold nanorods that mix together two coherent but orthogonally polarized light beams leading to modulation by an interference effect. By adjusting the phase and the amplitude of one of the beams, the amplitude and spectral composition of the second beam is altered. The plasmonic circuits display an asymmetry that enables an angle-dependent modulation, which we demonstrate with a diffraction grating where the energy directed into two diffraction orders is controlled by a second light beam. This effect appears like an optically controlled blaze that we use to switch a light beam between two different directions.


Drug Overdose Deaths Involving Cocaine and Psychostimulants with Abuse Potential - United States, 2003-2017.
In 2016, a total of 63,632 persons died from drug overdoses in the United States (1). Drug overdose deaths involving cocaine, psychostimulants with abuse potential (psychostimulants), or both substances combined increased 42.4% from 12,122 in 2015 to 17,258 in 2016.* Psychostimulants with abuse potential include drugs such as methamphetamine, 3,4-methylenedioxy-methamphetamine (MDMA), dextroamphetamine, levoamphetamine, methylphenidate (Ritalin), and caffeine. From 2015 to 2016, cocaine-involved and psychostimulant-involved death rates increased 52.4% and 33.3%, respectively (1). A total of 70,237 persons died from drug overdoses in the United States in 2017; approximately two thirds of these deaths involved an opioid (2). CDC analyzed 2016-2017 changes in age-adjusted death rates involving cocaine and psychostimulants by demographic characteristics, urbanization levels, U.S. Census region, 34 states, and the District of Columbia (DC). CDC also examined trends in age-adjusted cocaine-involved and psychostimulant-involved death rates from 2003 to 2017 overall, as well as with and without co-involvement of opioids. Among all 2017 drug overdose deaths, 13,942 (19.8%) involved cocaine, and 10,333 (14.7%) involved psychostimulants. Death rates increased from 2016 to 2017 for both drug categories across demographic characteristics, urbanization levels, Census regions, and states. In 2017, opioids were involved in 72.7% and 50.4% of cocaine-involved and psychostimulant-involved overdoses, respectively, and the data suggest that increases in cocaine-involved overdose deaths from 2012 to 2017 were driven primarily by synthetic opioids. Conversely, increases in psychostimulant-involved deaths from 2010 to 2017 occurred largely independent of opioids, with increased co-involvement of synthetic opioids in recent years. Provisional data from 2018 indicate that deaths involving cocaine and psychostimulants are continuing to increase.† Increases in stimulant-involved deaths are part of a growing polysubstance landscape. Increased surveillance and evidence-based multisectoral prevention and response strategies are needed to address deaths involving cocaine and psychostimulants and opioids. Enhancing linkage to care, building state and local capacity, and public health/public safety collaborations are critical components of prevention efforts.


Empirical treatment of Shigella dysentery with trimethoprim: five-day course vs. single dose.
Fifty-three adults hospitalized with Shigella dysentery were empirically treated with trimethoprim (200 mg) twice/day for 5 days, a single dose of trimethoprim (600 mg), or placebo in a randomized double-blind trial. During the first 24 hr of therapy, there was a reduction in the number of stools in 18/21 (86%) of patients treated with the 5-day regimen (trimethoprim-5) and 13/15 (87%) of patients treated with a single dose (trimethoprim-1), compared with 7/17 (41%) of the placebo group (P less than 0.025, both comparisons). The mean number of stools passed in the first 24 hr of therapy was 10.6, 10.8, and 21.3 stools in the trimethoprim-5, trimethoprim-1, and placebo groups, respectively. The mean (+/- SD) change in number of stools from baseline among treated patients during the first 24 hr was -4.9 (6.6) and -6.3 (6.3) for the trimethoprim-5 and trimethoprim-1 groups, respectively, compared with an increase of +2.4 (14.8) for the placebo group. There was a clinical failure at 48 hr in 9% of the trimethoprim-5 patients and 13% of trimethoprim-1 patients compared with 70% of placebo patients (P less than 0.005, both comparisons). Although we were unable to demonstrate a difference in efficacy between the two dosage schedules of trimethoprim, we conclude that both treatment regimens are effective for the treatment of Shigella dysentery.


Biosonar navigation above water II: exploiting mirror images.
As in vision, acoustic signals can be reflected by a smooth surface creating an acoustic mirror image. Water bodies represent the only naturally occurring horizontal and acoustically smooth surfaces. Echolocating bats flying over smooth water bodies encounter echo-acoustic mirror images of objects above the surface. Here, we combined an electrophysiological approach with a behavioral experimental paradigm to investigate whether bats can exploit echo-acoustic mirror images for navigation and how these mirrorlike echo-acoustic cues are encoded in their auditory cortex. In an obstacle-avoidance task where the obstacles could only be detected via their echo-acoustic mirror images, most bats spontaneously exploited these cues for navigation. Sonar ensonifications along the bats' flight path revealed conspicuous changes of the reflection patterns with slightly increased target strengths at relatively long echo delays corresponding to the longer acoustic paths from the mirrored obstacles. Recordings of cortical spatiotemporal response maps (STRMs) describe the tuning of a unit across the dimensions of elevation and time. The majority of cortical single and multiunits showed a special spatiotemporal pattern of excitatory areas in their STRM indicating a preference for echoes with (relative to the setup dimensions) long delays and, interestingly, from low elevations. This neural preference could effectively encode a reflection pattern as it would be perceived by an echolocating bat detecting an object mirrored from below. The current study provides both behavioral and neurophysiological evidence that echo-acoustic mirror images can be exploited by bats for obstacle avoidance. This capability effectively supports echo-acoustic navigation in highly cluttered natural habitats.


Effect of metformin and rosiglitazone in a prepubertal boy with Alström syndrome.
Alström syndrome (AS) is an autosomal recessive disorder characterized by progressive pigmentary retinopathy, sensorineural hearing loss, fatty liver infiltration, obesity, insulin resistance and early-onset type 2 diabetes mellitus (DM2). Early onset of insulin resistance and DM2 are key components of this syndrome. To study the effect of early initiation of the insulin sensitizer metformin combined with rosiglitazone in a patient with AS with impaired glucose tolerance. An 8 year-old boy with AS presented with acanthosis nigricans and insulin resistance at the age of 6 years. He had progressive excessive weight gain from 9 months of age. By the age of 1 year he developed photosensitivity, blindness and nystagmus. At the age of 5.5 years, his body mass index (BMI) was above the 95th percentile. He developed impaired glucose tolerance at 6 years of age and treatment with metformin was initiated. After 8 months of treatment with metformin he developed DM2. The dose of metformin was increased, and rosiglitazone added. A 2-hour oral glucose tolerance test (OGTT) and a rapid intravenous glucose tolerance test (IVGTT) were performed before treatment was initiated, after treatment with metformin and at the end of 1 year of combination therapy with metformin and rosiglitazone to calculate quantitative insulin sensitivity check index (QUICKI) and acute insulin response (AIR). For mutation analysis, all exons and splice site sequences of the ALMS1 gene were amplified and sequenced. Metformin treatment alone at the stage of impaired glucose tolerance did not prevent progression to DM2. However, metformin at a higher dose and in combination with rosiglitazone resulted in improvement of pancreatic beta-cell function, shown by markedly improved first-phase insulin response to glucose measured by AIR. The patient was found to have two heterozygous nonsense mutations in ALMS1, 8008 C-->T Ter, R2670X, and 11449 C-->T Ter, Q3817X. These alterations cause premature stops and result in a truncated ALMS1 protein. We suggest that early initiation of combined therapy comprising a high dose of metformin plus rosiglitazone may be valuable in managing insulin resistance and DM2 in children with AS.


The NK-1 receptor antagonist aprepitant as a broad spectrum antitumor drug.
Aprepitant is a selective high-affinity antagonist of human substance P (SP)/Neurokinin-1 (NK-1) receptors. Until now this drug has been used as anxiolytic, antidepressant and antiemetic. It has been demonstrated that SP induces cell proliferation and NK-1 receptor antagonists different to aprepitant inhibit growth in several human cancer cell lines, where NK-1 receptors are overexpressed. The purpose of this study is to demonstrate the antitumor action of aprepitant. We performed an in vitro study of the growth inhibition capacity of the NK-1 receptor antagonist aprepitant against glioma, neuroblastoma, retinoblastoma and pancreas, larynx, gastric and colon carcinomas cell lines. Coulter counter was used to determine viable cell numbers followed by application of the MTS colorimetric method. Furthermore, a DAPI method was applied to demonstrate apoptosis. We have demonstrated: aprepitant at (5-70 microM) concentration elicits growth cell inhibition in a concentration dependent manner in all tumor cell line studied. Maximum inhibition (100%) was observed when the aprepitant was administered at a concentration of > or = 70 microM in all tumor cell lines studied. The specific antitumor action of aprepitant occurs through the NK-1 receptor and tumor cells death was by apoptosis pathway. These findings reported here for the first time indicate that aprepitant is a new and promising broad spectrum antitumor drug in the treatment of cancer.


Clinical implications of hypertrophic cardiomyopathy associated with mutations in the alpha-tropomyosin gene.
The disease-bearing genes for hypertrophic cardiomyopathy (HCM) in HCM families have been identified as the beta-myosin heavy chain, alpha-tropomyosin, and cardiac troponin T genes. Three HCM kindreds with three distinct point mutations in the alpha-tropomyosin gene had extensive clinical evaluations. Single-strand conformation polymorphism gel analysis of polymerase chain reaction amplified products was used to capture each of the nine exons from the alpha-tropomyosin gene to identify mutations in 60 familial HCM patients. Two missense mutations in exon 2 (Ala63Val and Lys70Thr) and one missense mutation in exon 5 (Asp175Asn) were found in three unrelated HCM kindreds. These kindreds were the subject of clinical, electrocardiographic and echocardiographic studies. The morphological appearance of HCM was similar in the three kindreds. All the patients had severe hypertrophy of the left ventricle with asymmetrical septal hypertrophy during the early stage of the disease, which gradually progressed to dilatation of the left ventricle. Moreover, these kindreds showed similar disease penetrance, age of onset, and incidence of premature sudden death. The disease in these kindreds was severe and resulted in frequent sudden deaths. Among Japanese patients with familial HCM mutations in the alpha-tropomyosin gene are not as rare as reported, accounting for about 5% of all cases. These mutations are characterised by hypertrophy of the left ventricle which then progresses to dilatation and a high incidence of sudden or disease-related death.


Phase III randomized, double-blind study comparing single-dose intravenous peramivir with oral oseltamivir in patients with seasonal influenza virus infection.
Antiviral medications with activity against influenza viruses are important in controlling influenza. We compared intravenous peramivir, a potent neuraminidase inhibitor, with oseltamivir in patients with seasonal influenza virus infection. In a multinational, multicenter, double-blind, double-dummy randomized controlled study, patients aged ≥ 20 years with influenza A or B virus infection were randomly assigned to receive either a single intravenous infusion of peramivir (300 or 600 mg) or oral administration of oseltamivir (75 mg twice a day [b.i.d.] for 5 days). To demonstrate the noninferiority of peramivir in reducing the time to alleviation of influenza symptoms with hazard model analysis and a noninferiority margin of 0.170, we planned to recruit 1,050 patients in South Korea, Japan, and Taiwan. A total of 1,091 patients (364 receiving 300 mg and 362 receiving 600 mg of peramivir; 365 receiving oseltamivir) were included in the intent-to-treat infected population. The median durations of influenza symptoms were 78.0, 81.0, and 81.8 h in the groups treated with 300 mg of peramivir, 600 mg of peramivir, and oseltamivir, respectively. The hazard ratios of the 300- and 600-mg-peramivir groups compared to the oseltamivir group were 0.946 (97.5% confidence interval [CI], 0.793, 1.129) and 0.970 (97.5% CI, 0.814, 1.157), respectively. Both peramivir groups were noninferior to the oseltamivir group (97.5% CI, <1.170). The overall incidence of adverse drug reactions was significantly lower in the 300-mg-peramivir group, but the incidence of severe reactions in either peramivir group was not different from that in the oseltamivir group. Thus, a single intravenous dose of peramivir may be an alternative to a 5-day oral dose of oseltamivir for patients with seasonal influenza virus infection.


Comparison of traditional cardiovascular risk models and coronary atherosclerotic plaque as detected by computed tomography for prediction of acute coronary syndrome in patients with acute chest pain.
The objective was to determine the association of four clinical risk scores and coronary plaque burden as detected by computed tomography (CT) with the outcome of acute coronary syndrome (ACS) in patients with acute chest pain. The hypothesis was that the combination of risk scores and plaque burden improved the discriminatory capacity for the diagnosis of ACS. The study was a subanalysis of the Rule Out Myocardial Infarction Using Computer-Assisted Tomography (ROMICAT) trial-a prospective observational cohort study. The authors enrolled patients presenting to the emergency department (ED) with a chief complaint of acute chest pain, inconclusive initial evaluation (negative biomarkers, nondiagnostic electrocardiogram [ECG]), and no history of coronary artery disease (CAD). Patients underwent contrast-enhanced 64-multidetector-row cardiac CT and received standard clinical care (serial ECG, cardiac biomarkers, and subsequent diagnostic testing, such as exercise treadmill testing, nuclear stress perfusion imaging, and/or invasive coronary angiography), as deemed clinically appropriate. The clinical providers were blinded to CT results. The chest pain score was calculated and the results were dichotomized to ≥10 (high-risk) and <10 (low-risk). Three risk scores were calculated, Goldman, Sanchis, and Thrombolysis in Myocardial Infarction (TIMI), and each patient was assigned to a low-, intermediate-, or high-risk category. Because of the low number of subjects in the high-risk group, the intermediate- and high-risk groups were combined into one. CT images were evaluated for the presence of plaque in 17 coronary segments. Plaque burden was stratified into none, intermediate, and high (zero, one to four, and more than four segments with plaque). An outcome panel of two physicians (blinded to CT findings) established the primary outcome of ACS (defined as either an acute myocardial infarction or unstable angina) during the index hospitalization (from the presentation to the ED to the discharge from the hospital). Logistic regression modeling was performed to examine the association of risk scores and coronary plaque burden to the outcome of ACS. Unadjusted models were individually fitted for the coronary plaque burden and for Goldman, Sanchis, TIMI, and chest pain scores. In adjusted analyses, the authors tested whether the association between risk scores and ACS persisted after controlling for the coronary plaque burden. The prognostic discriminatory capacity of the risk scores and plaque burden for ACS was assessed using c-statistics. The differences in area under the receiver-operating characteristic curve (AUC) and c-statistics were tested by performing the -2 log likelihood ratio test of nested models. A p value <0.05 was considered statistically significant. Among 368 subjects, 31 (8%) subjects were diagnosed with ACS. Goldman (AUC = 0.61), Sanchis (AUC = 0.71), and TIMI (AUC = 0.63) had modest discriminatory capacity for the diagnosis of ACS. Plaque burden was the strongest predictor of ACS (AUC = 0.86; p < 0.05 for all comparisons with individual risk scores). The combination of plaque burden and risk scores improved prediction of ACS (plaque + Goldman AUC = 0.88, plaque + Sanchis AUC = 0.90, plaque + TIMI AUC = 0.88; p < 0.01 for all comparisons with coronary plaque burden alone). Risk scores (Goldman, Sanchis, TIMI) have modest discriminatory capacity and coronary plaque burden has good discriminatory capacity for the diagnosis of ACS in patients with acute chest pain. The combined information of risk scores and plaque burden significantly improves the discriminatory capacity for the diagnosis of ACS.


Discovery of dual function acridones as a new antimalarial chemotype.
Preventing and delaying the emergence of drug resistance is an essential goal of antimalarial drug development. Monotherapy and highly mutable drug targets have each facilitated resistance, and both are undesirable in effective long-term strategies against multi-drug-resistant malaria. Haem remains an immutable and vulnerable target, because it is not parasite-encoded and its detoxification during haemoglobin degradation, critical to parasite survival, can be subverted by drug-haem interaction as in the case of quinolines and many other drugs. Here we describe a new antimalarial chemotype that combines the haem-targeting character of acridones, together with a chemosensitizing component that counteracts resistance to quinoline antimalarial drugs. Beyond the essential intrinsic characteristics common to deserving candidate antimalarials (high potency in vitro against pan-sensitive and multi-drug-resistant Plasmodium falciparum, efficacy and safety in vivo after oral administration, inexpensive synthesis and favourable physicochemical properties), our initial lead, T3.5 (3-chloro-6-(2-diethylamino-ethoxy)-10-(2-diethylamino-ethyl)-acridone), demonstrates unique synergistic properties. In addition to 'verapamil-like' chemosensitization to chloroquine and amodiaquine against quinoline-resistant parasites, T3.5 also results in an apparently mechanistically distinct synergism with quinine and with piperaquine. This synergy, evident in both quinoline-sensitive and quinoline-resistant parasites, has been demonstrated both in vitro and in vivo. In summary, this innovative acridone design merges intrinsic potency and resistance-counteracting functions in one molecule, and represents a new strategy to expand, enhance and sustain effective antimalarial drug combinations.


Confusion with cerebral perfusion pressure in a literature review of current guidelines and survey of clinical practice.
Cerebral perfusion pressure (CPP) is defined as the difference between the mean arterial pressure (MAP) and the intracranial pressure (ICP). However, since patients with traumatic brain injury (TBI) are usually treated with head elevation, the recorded CPP values depends on the zero level used for calibration of the arterial blood pressure. Although international guidelines suggest that target values of optimal CPP are within the range of 50 - 70 mmHg in patients with TBI, the calibration of blood pressure, which directly influences CPP, is not described in the guidelines.The aim of this study was to review the literature used to support the CPP recommendations from the Brain Trauma Foundation, and to survey common clinical practice with respect to MAP, CPP targets and head elevation in European centres treating TBI patients. A review of the literature behind CPP threshold recommendations was performed. Authors were contacted if the publications did not report how MAP or CPP was measured. A short questionnaire related to measurement and treatment targets of MAP and CPP was sent to European neurosurgical centres treating patients with TBI. Assessment methods for CPP measurement were only retrieved from 6 of the 11 studies cited in the TBI guidelines. Routines for assessment of CPP varied between these 6 publications. The 58 neurosurgical centres that answered our survey reported diverging routines on how to measure MAP and target CPP values. Higher CPP threshold were not observed if blood pressure was calibrated at the heart level (p = 0.51). The evidence behind the recommended CPP thresholds shows no consistency on how blood pressure is calibrated and clinical practice for MAP measurements and CPP target values seems to be highly variable. Until a consensus is reached on how to measure CPP, confusion will prevail.


Peptide bonds affect the formation of haloacetamides, an emerging class of N-DBPs in drinking water: free amino acids versus oligopeptides.
Haloacetamides (HAcAms), an emerging class of nitrogenous disinfection by-products (N-DBPs) of health concern, have been frequently identified in drinking waters. It has long been appreciated that free amino acids (AAs), accounting for a small fraction of the dissolved organic nitrogen (DON) pool, can form dichloroacetamide (DCAcAm) during chlorination. However, the information regarding the impacts of combined AAs, which contribute to the greatest identifiable DON portion in natural waters, is limited. In this study, we compared the formation of HAcAms from free AAs (tyrosine [Tyr] and alanine [Ala]) and combined AAs (Tyr-Ala, Ala-Tyr, Tyr-Tyr-Tyr, Ala-Ala-Ala), and found that HAcAm formation from the chlorination of AAs in combined forms (oligopeptides) significantly exhibited a different pattern with HAcAm formation from free AAs. Due to the presence of peptide bonds in tripeptides, Tyr-Tyr-Tyr and Ala-Ala-Ala produced trichloroacetamide (TCAcAm) in which free AAs was unable to form TCAcAm during chlorination. Moreover, peptide bond in tripeptides formed more tri-HAcAms than di-HAcAms in the presence of bromide. Therefore, the peptide bond may be an important indicator to predict the formation of specific N-DBPs in chlorination. The increased use of algal- and wastewater-impacted water as drinking water sources will increase health concerns over exposure to HAcAms in drinking water.


Type 2 Diabetes Prevention Diet and Hepatocellular Carcinoma Risk in US Men and Women.
Adherence to a healthy diet has been associated with a reduced risk of type 2 diabetes (T2D). Hepatocellular carcinoma (HCC) may have overlapping mechanisms with T2D, such as inflammation and insulin resistance. Thus, we examined the association between a previously developed T2D prevention dietary pattern and HCC risk. We followed 87,943 women in the Nurses' Health Study and 49,665 men in the Health Professionals Follow-up Study for up to 32 years. The dietary diabetes risk reduction score, which includes dietary glycemic index, cereal fiber, ratio of polyunsaturated to saturated fats, trans fat, sugar-sweetened beverages, nuts, coffee, and red and processed meats, was obtained using validated food frequency questionnaires and updated every 4 years. The Cox proportional hazards regression model was used to calculate multivariable hazard ratios and confidence intervals (95% CIs). During over 1.9 million person-years, a total of 160 incident HCC cases were identified. The dietary diabetes risk reduction score was associated with a lower risk of HCC (top vs bottom quartile; hazard ratio: 0.57, 95% CI: 0.34-0.95; Ptrend = 0.03). All the individual food and beverage items were associated with the risk of HCC in the expected direction, although the association was weaker than the overall dietary pattern. Greater adherence to the T2D prevention diet was associated with a lower risk of developing HCC among US men and women. Further studies are needed to confirm and extend our findings.


[Migraine and irritable bowel syndrome].
The association between migraine and functional gastrointestinal disorders has been confirmed by many clinical observations and epidemiological studies. In most patients during the attacks of migraine, apart from various neurological and vascular symptoms, gastrointestinal disturbances occur including nausea, vomiting, abdominal pain or diarrhea. Functional gastrointestinal disorders, such as irritable bowel syndrome (IBS), are reported in migraine patients in periods between the attacks as well. On the other hand 23-53% of IBS patients have frequent headaches. Migraine and IBS often coexist with fibromyalgia and other chronic pain syndromes and functional disorders. Migraine and IBS affect approximately 10-20% of the general population, usually young adults. Both diseases are more prevalent in women, perhaps due to the role of estrogen in their pathogenesis. Looking for the common pathogenetic mechanisms of IBS and migraine the role of the brain-gut axis, neuroimmune and neuroendocrine interactions are being considered. The influence of stress on symptom occurrence and severity seems to be associated with hyperactivity of the hypothalamic-pituitary-adrenal axis. The enteric nervous system as a source of numerous neurotransmitters and visceral reflexes is a plausible common pathogenic link between IBS and migraine. In particular serotonin being the main neurotransmitter of the gastrointestinal tract plays a relevant role in the pathogenesis of IBS as well as migraine. Nowadays, agonists and antagonists of serotoninergic receptors are the most efficacious drugs for IBS and migraine therapy. Some side effects of triptans, 5-HT(1B/D) agonists, used in migraine treatment may be connected with the influence of triptans on the gastrointestinal functions. A better understanding of the relationship between migraine and IBS may result in more effective treatment of both diseases.


Extraction of intrinsic fluorescence from single fiber fluorescence measurements on a turbid medium: experimental validation.
The detailed mechanisms associated with the influence of scattering and absorption properties on the fluorescence intensity sampled by a single optical fiber have recently been elucidated based on Monte Carlo simulated data. Here we develop an experimental single fiber fluorescence (SFF) spectroscopy setup and validate the Monte Carlo data and semi-empirical model equation that describes the SFF signal as a function of scattering. We present a calibration procedure that corrects the SFF signal for all system-related, wavelength dependent transmission efficiencies to yield an absolute value of intrinsic fluorescence. The validity of the Monte Carlo data and semi-empirical model is demonstrated using a set of fluorescent phantoms with varying concentrations of Intralipid to vary the scattering properties, yielding a wide range of reduced scattering coefficients (μ's = 0-7 mm (-1)). We also introduce a small modification to the model to account for the case of μ's = 0 mm (-1) and show its relation to the experimental, simulated and theoretically calculated value of SFF intensity in the absence of scattering. Finally, we show that our method is also accurate in the presence of absorbers by performing measurements on phantoms containing red blood cells and correcting for their absorption properties.


Sex differences in youth-reported depressive symptomatology and unwanted internet sexual solicitation.
As the number of youths using the Internet regularly increase, so too does the number of youths potentially vulnerable to negative experiences online. Clinicians, policy makers, and parents need to better understand the Internet and factors related to positive and negative experiences online. Primarily to investigate the association between youth-reported depressive symptomatology and unwanted Internet sexual solicitation and secondarily to identify sex differences in related characteristics of affected youth. Data from the Youth Internet Safety Survey were analyzed to investigate the association between reported depressive symptomatology and unwanted Internet sexual solicitation. The Youth Internet Safety Survey was a nationally-representative, cross-sectional telephone survey. Youth participants (N = 1501) were English speakers between the ages of 10 and 17 years who had accessed the Internet at least 6 times in the previous 6 months and had resided in the household for at least 2 weeks in the previous year. Eighty-two percent of contacted households agreed to participate. Each participant was asked to indicate whether any of the 9 symptoms of major depression defined by the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) had been present within the previous month. Logistic regression was used to estimate the odds of reporting an unwanted sexual solicitation online for youths with mild or major depressive symptomatology versus no symptomatology. A parsimonious, multivariate model of significant youth characteristics was identified separately for males and females. Youths who report major depressive-like symptoms were 3.5 times more likely (odds ratio, 3.54; 95% confidence interval, 2.19-5.71) to also report an unwanted sexual solicitation online compared to youths with mild/no symptomatology. After adjusting for significant Internet and psychosocial characteristics, male Internet users who report major depressive-like symptomatology were 2.5 times more likely to also indicate an unwanted Internet solicitation (adjusted odds ratio, 2.72; 95% CI, 1.15-6.40); significant differences were not observed among otherwise-similar females. Further, among youths reporting an Internet solicitation (N = 283), youths with major depressive-like symptomatology were twice as likely to report feeling emotionally distressed by the incident compared to youths with mild/no symptomatology (odds ratio, 2.27; 95% CI, 1.03-5.02). While the majority of youths report positive experiences online, some youths may be more vulnerable to negative experiences. Cross-sectional results indicate that the report of depressive symptomatology is significantly related to the concurrent report of an unwanted Internet sexual solicitation, especially for young males. Future research should focus on parsing out the temporality of events and identifying additional populations of vulnerable youths online.


Hybrid Continuous-Flow Total Artificial Heart.
Clinical studies using total artificial hearts (TAHs) have demonstrated that pediatric and adult patients derive quality-of-life benefits from this form of therapy. Two clinically-approved TAHs and other pumps under development, however, have design challenges and limitations, including thromboembolic events, neurologic impairment, infection risk due to large size and percutaneous drivelines, and lack of ambulation, to name a few. To address these limitations, we are developing a hybrid-design, continuous-flow, implantable or extracorporeal, magnetically-levitated TAH for pediatric and adult patients with heart failure. This TAH has only two moving parts: an axial impeller for the pulmonary circulation and a centrifugal impeller for the systemic circulation. This device will utilize the latest generation of magnetic bearing technology. Initial geometries were established using pump design equations, and computational modeling provided insight into pump performance. The designs were the basis for prototype manufacturing and hydraulic testing. The study results demonstrate that the TAH is capable of delivering target blood flow rates of 1-6.5 L/min with pressure rises of 1-92 mm Hg for the pulmonary circulation and 24-150 mm Hg for the systemic circulation at 1500-10 000 rpm. This initial design of the TAH was successful and serves as the foundation to continue its development as a novel, more compact, nonthrombogenic, and effective therapeutic alternative for infants, children, adolescents, and adults with heart failure.


A mass balance of tri-hexabrominated diphenyl ethers in lactating cows.
Beef and dairy products can be important vectors of human exposure to polybrominated diphenylethers (BDEs), and hence an understanding of BDE transfer from feed to cows' milk and tissue is important for BDE exposure assessment The fate of tri- to hexaBDEs in lactating cows exposed to a naturally contaminated diet was studied by analyzing feed, feces, and milk samples from a mass balance study. Tissue distribution was studied in one cowslaughtered afterthe experiment The carryover rates from feed to milk ranged from 0.15 to 0.35 for the major congeners. Lower values were observed for several of the tetrabrominated congeners, and this was attributed to metabolism. The dietary absorption efficiency decreased with increasing octanol-water partition coefficient of the BDE congener. The absorption behavior was consistent with a model based on chemical lipophilicity, but agreed less well with a model based on effective molecular diameter, and it violated Lipinski's "rule of 5". The lipid normalized concentrations were similar in all tissues analyzed including liver and milk, suggesting that tissue distribution is governed by partitioning into lipids. Overall, the behavior of the tri- to hexaBDEs was consistent with that observed for other classes of halogenated aromatic contaminants such as PCBs and PCDD/Fs, but it differed markedly from the behavior of the hepta- decaBDEs.


Bowel cancer screening in England: a qualitative study of GPs' attitudes and information needs.
The National Health Service Bowel Cancer Screening Programme is to be introduced in England during 2006. General Practitioners are a potentially important point of contact for participants throughout the screening process. The aims of the study were to examine GPs' attitudes and information needs with regard to bowel cancer screening, with a view to developing an information pack for primary care teams that will be circulated prior to the introduction of the programme. 32 GPs participated in semi-structured telephone interviews. 18 of these had participated in the English Bowel Screening Pilot, and 14 had not. Interviews covered attitudes towards the introduction of the Bowel Cancer Screening Programme, expected or actual increases in workload, confidence in promoting informed choice, and preferences for receiving information about the programme. GPs in the study were generally positive about the introduction of the Bowel Cancer Screening Programme. A number of concerns were identified by GPs who had not taken part in the pilot programme, particularly relating to patient welfare, patient participation, and increased workload. GPs who had taken part in the pilot reported holding similar concerns prior to their involvement. However, in many cases these concerns were not confirmed through GPs experiences with the pilot. A number of specific information needs were identified by GPs to enable them to provide a supportive role to participants in the programme. The study has found considerable GP support for the introduction of the new Bowel Cancer Screening Programme. Nonetheless, GPs hold some significant reservations regarding the programme. It is important that the information needs of GPs and other members of the primary care team are addressed prior to the roll-out of the programme so they are equipped to promote informed choice and provide support to patients who consult them with queries regarding screening.


Laryngeal cancer cost analysis: association of case-mix and treatment characteristics with medical charges.
To examine the relationship of various pretreatment case-mix characteristics and treatment modalities with medical charges incurred during diagnosis, treatment, and 2-year follow-up for patients with laryngeal cancer. Retrospective chart review and billing record analysis. The charts and billing records of patients diagnosed with laryngeal cancer at the University of Iowa Hospitals and Clinics (UIHC) between January 1, 1991 and December 31, 1994 were reviewed. The independent variables included various pretreatment patient-mix and tumor characteristics (age, AJCC TNM clinical stage, smoking history, ASA class, and comorbidity as defined by Kaplan-Feinstein grade) as well as type of treatment. The dependent variables included total physician, office, and university hospital-based charges incurred during the pretreatment evaluation and 0- to 3-, 3- to 12, and 12- to 24-month billing periods after the initiation of cancer-directed therapy. Total 1-year and 2-year charges were also evaluated. Univariate and multivariate analyses were used to investigate the relationships between dependent and independent variables and to develop models predictive of management charges during the individual and total billing periods. Pretreatment charges showed no significant associations (P < .05) with any of the independent variables. Multiple regression analyses indicated that comorbidity, stage, and initial treatment modality were significant variables in one or more of the models predicting charges incurred during the 0- to 3-month, 3- to 12-month, total 1-year, and total 2-year billing periods. The models yielded R2 values for the total 1- and 2-year billing periods of 0.5246 and 0.5055, respectively. This work supports continued study of measures that may result in earlier detection of laryngeal cancer as a potential means of reducing management charges. These results also indicate that a more accurate method of stratifying the disease severity of laryngeal cancer patients for reimbursement purposes would include measurements of the severity of the index disease as well as comorbid diseases.


Review of mortality of very low birthweight infants at the University Hospital of the West Indies over the past four decades.
A review of two previously published studies done at the University Hospital of the West Indies, an unpublished study and annual perinatal statistics was conducted to detect trends in the mortality of very low birthweight infants at the institution over four decades. Mortality decreased from 54% to 38% over the time period, the decrease was greater for infants weighing 1001-1500 g (40%) than those weighing < or = 1000 g (28%). Despite increased access to mechanical ventilation over time, there was no appreciable decrease in mortality for infants weighing < or = 750 g. There was a statistically significant decrease in mortality with increasing birthweight for the time period 1987-2002, p < 0.001. The mean +/- SD weight of survivors 1.18 +/- 0.24 kg was significantly greater than that for non-survivors 0.89 +/- 0.21 kg for the same period. Further decrease in mortality of very low birthweight infants will involve measures aimed at decreasing mortality in infants weighing < or = 750 g and increasing the availability of parenteral nutrition and the accessibility of surfactant.


Transplantation of allogenic bone marrow in canine cyclic neutropenia.
Transplantation of normal bone marrow cells to a gray collie dog with cyclic neutropenia resulted in normal granulocytopoiesis. The finding suggests that cyclic neutropenia occurs because the hematopoietic stem cells are defective. Because of the similarity of human and canine cyclic neutropenia, it also suggests that the human disease may be curable by marrow transplantation.


Long-Range Orientational Self-Assembly, Spatially Controlled Deprotonation, and Off-Centered Metalation of an Expanded Porphyrin.
Expanded porphyrins are large-cavity macrocycles with enormous potential in coordination chemistry, anion sensing, photodynamic therapy, and optoelectronics. In the last two decades, the surface science community has assessed the physicochemical properties of tetrapyrrolic-like macrocycles. However, to date, the sublimation, self-assembly and atomistic insights of expanded porphyrins on surfaces have remained elusive. Here, we show the self-assembly on Au(111) of an expanded aza-porphyrin, namely, an "expanded hemiporphyrazine", through a unique growth mechanism based on long-range orientational self-assembly. Furthermore, a spatially controlled "writing" protocol on such self-assembled architecture is presented based on the STM tip-induced deprotonation of the inner protons of individual macrocycles. Finally, the capability of these surface-confined macrocycles to host lanthanide elements is assessed, introducing a novel off-centered coordination motif. The presented findings represent a milestone in the fields of porphyrinoid chemistry and surface science, revealing a great potential for novel surface patterning, opening new avenues for molecular level information storage, and boosting the emerging field of surface-confined coordination chemistry involving f-block elements.


Dosage compensation in mammals.
Many organisms show major chromosomal differences between sexes. In mammals, females have two copies of a large, gene-rich chromosome, the X, whereas males have one X and a small, gene-poor Y. The imbalance in expression of several hundred genes is lethal if not dealt with by dosage compensation. The male-female difference is addressed by silencing of genes on one female X early in development. However, both males and females now have only one active X chromosome. This is compensated by twofold up-regulation of genes on the active X. This complex system continues to provide important insights into mechanisms of epigenetic regulation.


Purpura haemorrhagica in 53 horses.
The medical records of 53 horses with purpura haemorrhagica were reviewed. Seventeen of them had been exposed to or infected with Streptococcus equi, nine had been infected with Corynebacterium pseudotuberculosis, five had been vaccinated with S. equi M protein, five had had a respiratory infection of unknown aetiology, and two had open wounds; the other 15 cases had no history of recent viral or bacterial infection. The horses were between six months and 19 years of age (mean 8.4 years). The predominant clinical signs were well demarcated subcutaneous oedema of all four limbs and haemorrhages on the visible mucous membranes; other signs included depression, anorexia, fever, tachycardia, tachypnoea, reluctance to move, drainage from lymph nodes, exudation of serum from the skin, colic, epistaxis and weight loss. Haematological and biochemical abnormalities commonly detected were anaemia, neutrophilia, hyperproteinaemia, hyperfibrinogenaemia, hyperglobulinaemia and high activities of muscle enzymes. All of the horses were treated with corticosteroids; 42 also received non-steroidal anti-inflammatory drugs and 26 received antimicrobial drugs. Selected cases received special nursing care, including hydrotherapy and bandaging of the limbs. Most of the horses were treated for more than seven days and none of them relapsed. Forty-nine of the horses survived, one died and three were euthanased, either because their severe clinical disease failed to respond to treatment or because they developed secondary complications. Two of the four non-survivors had been vaccinated against S. equi with a product containing the M protein, one had a S. equi infection and the other had a respiratory infection of undetermined aetiology.


Abdominal tuberculosis: a histopathological study with special reference to intestinal perforation and mesenteric vasculopathy.
Along with the increased incidence of pulmonary tuberculosis in parallel with the increase in population in various parts of the world, in recent years, the incidence of abdominal tuberculosis has also increased. The pathogenetic events in intestinal tuberculosis, which culminate in ulcer formation, perforation, and stricture, still have to be identified. To correlate the gross and microscopic features in intestinal tuberculosis, in particular tuberculous perforation with changes in mesenteric vasculature. A one-year prospective study of excised/resected tissues from patients with abdominal tuberculosis requiring surgical intervention was conducted. Tissues from fifty-six patients were included in the study-of which 36 were resected intestinal segments and 20 were intestinal and lymph node biopsies. Hematoxylin and Eosin and Ziehl-Neelsen stains were used for histopathological examination. Tuberculous enteritis was found to be present in 49 of the 56 patients (87.5%) (ileum being the site most commonly affected), while nodal involvement was seen in 39 (69.6%) patients. Perforations were present in 39 out of 49 (79.6%) intestinal tissues; most being solitary and ileum was the commonest site. Typical epithelioid cell granulomas were seen in the intestine and lymph nodes, with caseation being more prevalent in the latter. The mesenteric vasculature was frequently involved by granulomatous inflammation, with intravascular organizing thrombus being present in 30% of the resected specimens with perforation. Acid fast bacilli were demonstrated in the tissue sections of 37.5% of the patients. AFB positivity was higher in caseating granulomas. Involvement of mesenteric vasculature by granulomatous inflammation was commonly associated with the ulcerative type with perforation, suggesting that ischemia caused by vascular thrombosis is responsible for tissue breakdown. This implies that vasculitis plays an important role in the natural history of abdominal tuberculosis.


Conservation Genetics of the Scalloped Hammerhead Shark in the Pacific Coast of Colombia.
Previous investigations of the population genetics of the scalloped hammerhead sharks (Sphyrna lewini) in the Eastern Tropical Pacific have lacked information about nursery areas. Such areas are key to promoting conservation initiatives that can protect young sharks from threats such as overfishing. Here, we investigated the genetic diversity, phylogeography, and connectivity of S. lewini found in 3 areas of Colombia's Pacific coast: around Malpelo Island and in 2 National Natural Parks on the Colombian Pacific mainland (Sanquianga and Ensenada de Utría). We analyzed mtDNA control region (CR) sequences and genotyped 15 microsatellite loci in 137 samples of adults and juveniles. The mtDNA analyses showed haplotypes shared between the Colombian Pacific individuals sampled in this investigation and other areas in the Eastern Tropical Pacific, the Indo-Pacific, and with sequences previously reported in Colombia (Buenaventura Port), as well as 4 unique haplotypes. Population assignment and paternity analyses detected 3 parent-offspring pairs between Malpelo and Sanquianga and 1 between Malpelo and Utría. These results indicate high genetic connectivity between Malpelo Island and the Colombian Pacific coast, suggesting that these 2 areas are nurseries for S. lewini. This is, to our knowledge, the first evidence of nursery areas identified for the scalloped hammerhead shark anywhere in the world. Additional conservation planning may be required to protect these nursery habitats of this endangered shark species.


Nonlinear Evolution and Final Fate of Charged Anti-de Sitter Black Hole Superradiant Instability.
We describe the full nonlinear development of the superradiant instability for a charged massless scalar field coupled to general relativity and electromagnetism, in the vicinity of a Reissner-Nordström-anti-de Sitter black hole. The presence of the negative cosmological constant provides a natural context for considering perfectly reflecting boundary conditions and studying the dynamics as the scalar field interacts repeatedly with the black hole. At early times, small superradiant perturbations grow as expected from linearized studies. Backreaction then causes the black hole to lose charge and mass until the perturbation becomes nonsuperradiant, with the final state described by a stable hairy black hole. For large gauge coupling, the instability extracts a large amount of charge per unit mass, resulting in greater entropy increase. We discuss the implications of the observed behavior for the general problem of superradiance in black hole spacetimes.


Children and Adults Both Learn Motor Sequences Quickly, But Do So Differently.
Both children and adults can learn motor sequences quickly in one learning session, yet little is known about potential age-related processes that underlie this fast sequence acquisition. Here, we examined the progressive performance changes in a one-session modified serial reaction time task in 6- and 10-year-old children and adults. We found that rapid sequence learning, as reflected by reaction time (RT), was comparable between groups. The learning was expressed through two behavioral processes: online progressive changes in RT while the task was performed in a continuous manner and offline changes in RT that emerged following a short rest. These offline and online RT changes were age-related; learning in 6-year-olds was primarily reflected through the offline process. In contrast, learning in adults was reflected through the online process; and both online and offline processes occurred concurrently in 10-year-olds. Our results suggest that early rapid sequence learning has a developmental profile. Although the unifying mechanism underlying these two age-related processes is unclear, we discuss possible explanations that need to be systematically elucidated in future studies.


Silicon balance in human volunteers; a pilot study to establish the variance in silicon excretion versus intake.
Accumulating evidence suggests a role for silicon in optimal connective tissue health. Further proof of its importance/essentiality may be provided by studies involving imposed depletion followed by 29Si challenge to estimate metabolic balance. Prior to conducting these expensive studies, we first established the variance of estimating normal Si excretion versus intake using a single oral dose of typical dietary Si, orthosilicic acid. Healthy volunteers were recruited from Loei Rajabhat University, separated into two matched groups (three males and three females/group) and maintained on a standardized diet for the three study days. One group ingested 500 ml water containing orthosilicic acid (28.9 mg Si) and the other group received 500 ml water alone, all on a fasted stomach. Blood samples and total urine and faeces were collected over the 48 h post-dose period and 24 h before-hand (baseline) and analysed for silicon by inductively coupled plasma optical emission spectrometry. Serum Si analysis confirmed the ready absorption of silicon from the orthosilicic acid solution. Mean total urinary and faecal Si excretions over the 24 h post-dose period accounted for 57 ± 9.5% and 39 ± 9.4% of the ingested dose, respectively. Thus in total 96.3 ± 5.8% of the ingested dose was recovered in faecal plus urinary excretions over the 24 h post-dose period. We report that in healthy subjects (presumably in Si balance), the ingestion of a soluble dose of dietary Si results in the same quantity (within analytical error) being excreted within 24 h. It is currently not known if this all originated from the dose solution or if there was some exchange with the body Si pool but, given the low variance in these silicon balance data, isotopic studies are now merited.


Expression of gene rrg is associated with reversion of NIH 3T3 transformed by LTR-c-H-ras.
A partial complementary DNA was isolated for a gene (rrg) that is normally expressed in mouse NIH 3T3 cells, but is down-regulated after cellular transformation by long terminal repeat (LTR)-activated c-H-ras (LTR-c-H-ras). This gene was reexpressed in a nontumorigenic persistent revertant cell line created by prolonged treatment of the transformed cells with mouse interferon alpha/beta. Persistent revertants stably transfected with rrg complementary DNA antisense expression vectors appeared transformed, had decreased amounts of rrg messenger RNA, and were tumorigenic in nude mice. Stable transfection with sense constructs did not alter the normal morphology, message level, or nontumorigenicity of the persistent revertant cell line.


Splenic and lung response to nonlethal systemic Aspergillus fumigatus infection in C57BL/6 mice.
In this study, we investigated splenic and lung cell responses to nonlethal systemic Aspergillus fumigatus infection in mice. Apart from basic indices of spleen and lung cell activity, IL-17 expression by cells from both tissues was determined and compared to the expression of IFN-gamma and IL-4. Progressive decrease in tissue fungal burden correlated with increased spleen and lung cell activity. Increased IL-17 message was noted in spleen cells at all time points (3, 7 and 15 days post-infection; p.i.), while a modest increase in IFN-gamma mRNA expression was noted at day 3 p.i. Increased cytokine production at days 3 and 7 (IL-17) and throughout the experimental period (IFN-gamma) was found. In contrast, spleen cell IL-4 expression was considerably lower during infection, resulting in high IFN-gamma/IL-4 and IL-17/IL-4 ratios in the spleen. Pro-inflammatory cytokine response was observed in the lungs as well, but primarily as the result of increased production of IFN-gamma by lung cells in response to challenge with conidia and the absence of change in IL-4 response. Increased activity of cells from both tissues, as well as the pattern of cytokine production, created an optimal pro-inflammatory milieu for fungal eradication.


Learning to read improves the structure of the arcuate fasciculus.
The acquisition of literacy results from an effortful learning process that leads to functional changes in several cortical regions. We explored whether learning to read also leads to anatomical changes within the left intrahemispheric white matter pathways that interconnect these regions. Using diffusion tensor imaging tractography, we compared illiterates with ex-illiterates who learned to read during adulthood and literates who learned to read during their childhood. Literacy related to an increase in fractional anisotropy and a decrease in perpendicular diffusivity in the temporo-parietal portion of the left arcuate fasciculus. The microstructure within this pathway correlated with the reading performance and the degree of functional activation within 2 dominant brain regions involved in reading: The Visual Word Form Area in response to letter strings, and the posterior superior temporal cortex in response to spoken language. Thus, the acquisition of literacy is associated with a reinforcement of left temporo-parietal connections whose microstructure predicts overall reading performance and the functional specialization of the Visual Word Form Area. This anatomical magnetic resonance imaging marker may be useful to predict developmental reading disorders.


Lunatic, Manic, and Radical Fringe Each Promote T and B Cell Development.
Lunatic, Manic, and Radical Fringe (LFNG, MFNG, and RFNG) are N-acetylglucosaminyltransferases that modify Notch receptors and regulate Notch signaling. Loss of LFNG affects thymic T cell development, and LFNG and MFNG are required for marginal zone (MZ) B cell development. However, roles for MFNG and RFNG in T cell development, RFNG in B cell development, or Fringes in T and B cell activation are not identified. In this study, we show that Lfng/Mfng/Rfng triple knockout (Fng tKO) mice exhibited reduced binding of DLL4 Notch ligand to CD4/CD8 double-negative (DN) T cell progenitors, and reduced expression of NOTCH1 targets Deltex1 and CD25. Fng tKO mice had reduced frequencies of DN1/cKit(+) and DN2 T cell progenitors and CD4(+)CD8(+) double-positive (DP) T cell precursors, but increased frequencies of CD4(+) and CD8(+) single-positive T cells in the thymus. In spleen, Fng tKO mice had reduced frequencies of CD4(+), CD8(+), central memory T cells and MZ B cells, and an increased frequency of effector memory T cells, neutrophils, follicular, and MZ P B cells. The Fng tKO phenotype was cell-autonomous and largely rescued in mice expressing one allele of a single Fng gene. Stimulation of Fng tKO splenocytes with anti-CD3/CD28 beads or LPS gave reduced proliferation compared with controls, and the generation of activated T cells by Concanavalin A or L-PHA was also reduced in Fng tKO mice. Therefore, each Fringe contributes to T and B cell development, and Fringe is required for optimal in vitro stimulation of T and B cells.


Which long-acting bronchodilator is most cost-effective for the treatment of COPD?
The aim of this study was to estimate the cost-effectiveness of tiotropium versus salmeterol to inform decision making within the Dutch healthcare setting. A previously published, validated COPD progression model was updated with new exacerbation data and adapted to the Dutch setting by including Dutch estimates of healthcare use for COPD maintenance treatment and Dutch unit costs. Exacerbation data from the POET-COPD trial were combined with evidence from earlier tiotropium studies using Bayesian meta-analysis. The model-based analysis was performed using a one- and five-year time horizon. Main health outcomes were the number of exacerbations and quality-adjusted life years (QALYs). One-year costs per patient from the healthcare perspective were v1370 for tiotropium and v1359 for salmeterol; a difference of v11 (95% uncertainty interval (UI): -198-212). The annual number of exacerbations was 0.068 (-0.005-0.140) lower in the tiotropium group. The number of QALYs in the tiotropium group was 0.011 (-0.019-0.049) higher, resulting in an incremental cost-effectiveness ratio (ICER) of v1015 per QALY. After five years, the difference in exacerbations, QALYs and costs between the tiotropium and salmeterol group were -0.435 (-0.915-0.107), 0.079 (-0.272-0.520) and v-277 (-1586-1074), respectively, indicating that tiotropium was more effective and less costly. Using a societal perspective, tiotropium dominated salmeterol both after one and five years. Tiotropium reduced exacerbations and exacerbation-related costs. After one year the cost per QALY of tiotropium compared with salmeterol was very low, while after five years tiotropium was found to dominate salmeterol.


Proteomic analysis of synaptosomes using isotope-coded affinity tags and mass spectrometry.
Synaptosomes are isolated synapses produced by subcellular fractionation of brain tissue. They contain the complete presynaptic terminal, including mitochondria and synaptic vesicles, and portions of the postsynaptic side, including the postsynaptic membrane and the postsynaptic density (PSyD). A proteomic characterisation of synaptosomes isolated from mouse brain was performed employing the isotope-coded affinity tag (ICAT) method and tandem mass spectrometry (MS/MS). After isotopic labelling and tryptic digestion, peptides were fractionated by cation exchange chromatography and cysteine-containing peptides were isolated by affinity chromatography. The peptides were identified by microcapillary liquid chromatography-electrospray ionisation MS/MS (muLC-ESI MS/MS). In two experiments, peptides representing a total of 1131 database entries were identified. They are involved in different presynaptic and postsynaptic functions, including synaptic vesicle exocytosis for neurotransmitter release, vesicle endocytosis for synaptic vesicle recycling, as well as postsynaptic receptors and proteins constituting the PSyD. Moreover, a large number of soluble and membrane-bound molecules serving functions in synaptic signal transduction and metabolism were detected. The results provide an inventory of the synaptic proteome and confirm the suitability of the ICAT method for the assessment of synaptic structure, function and plasticity.


Distinct orbitofrontal regions encode stimulus and choice valuation.
The weak axiom of revealed preferences suggests that the value of an object can be understood through the simple examination of choices. Although this axiom has driven economic theory, the assumption of equation between value and choice is often violated. fMRI was used to decouple the processes associated with evaluating stimuli from evaluating one's actions. Whereas activity in left posterior areas of the orbitofrontal cortex (OFC) was associated with processing the objective value of stimuli, activity in medial anterior areas of the OFC was associated with accepting high value gambles and rejecting low value gambles; that is, making correct decisions. These data demonstrate that distinct areas of the OFC provide dissociated representations for use in adaptive decision-making and suggest an important processing distinction between the concepts of good/bad and right/wrong.


Specific fiber deficits in sensorimotor diabetic polyneuropathy correspond to cytotoxicity against neuroblastoma cells of sera from patients with diabetes.
Neuropathy is the most common complication of diabetes, and toxic serum factors may contribute to its genesis. We assessed neurotoxicity in the serum of 39 diabetic patients and correlated it with clinical measures of somatic and autonomic nerve fiber damage. Sera were applied to N1E-115 and VSC4.1 neuroblastoma cells in vitro as models of sensory/autonomic (S/A) and motor neurons, respectively. Neurotoxicity was measured as either complete or near-complete cell death (highly toxic), inhibited cell growth (moderately toxic), or normal cell proliferation (nontoxic) compared with pooled human serum controls during culture over 4 days. There was an inverse correlation between neurotoxicity and vibration perception threshold (P < 0.01). Age (P < 0.02), duration of diabetes (P < 0.02), and HbA1c (P < 0.03) correlated with neurotoxicity, suggesting that glycation may contribute to cytotoxicity in this model. S/A neurotoxicity occurred more frequently in the sera of patients with type 1 (19 of 25) than type 2 (5 of 14) diabetes (P < 0.02). None of the sera from either type 1 or type 2 diabetic patients displayed neurotoxicity on VSC4.1 cells, whereas sera from patients with motor neuropathy were highly toxic. These studies indicate that there is a relationship between the specific nerve fiber dysfunction in the patient and the type of neuronal cell killed, not only for diabetic neuropathy but also for known forms of autoimmune neuropathies. Such toxic factors may contribute to diabetic neuropathy by acting in concert with hyperglycemia to damage sensory/autonomic neurons.


The efficacy of immediate diet for reducing local adverse events of inhaled corticosteroid: a pilot study.
Local adverse events associated with inhaled corticosteroid use, including dysphonia, pharyngitis and oral candidiasis, can affect adherence for treatment. 'Mouth rinsing method' has been used for reducing local adverse events, but it cannot ensure complete prevention. The goal of this pilot study was to identify whether the 'immediate diet method' can reduce local adverse events in a limited number of patients. The study was conducted in a total of 98 patients, who had been prescribed a medium-dose fluticasone propionate for the first time, from January to October in 2010. One training nurse had performed the education on how to use the inhaler, including the mouth rinsing method. And with follow-ups at one month intervals, any patient who experienced such adverse events were educated on the immediate diet method, having a meal within 5 minutes after using an inhaler and they were checked on any incurrence of adverse events with one month intervals for 2 months. The mean age of patients was 65.9 years old. The local adverse events had incurred from 18.4% of the study subjects. When performed the follow-up observation in 18 patients with local adverse events after education on the immediate diet method, 14 patients (77.8%) had shown symptomatic improvements. Three of 4 patients did not show any improvement, in spite of implementing the immediate diet method. The other 1 patient did not practice the immediate diet method properly. The immediate diet method may be useful in reducing the local adverse events, caused by the use of inhaled corticosteroid.


A diffusion tensor imaging study on the white matter skeleton in individuals with sports-related concussion.
Recognizing and managing the effects of cerebral concussion is very challenging, given the discrete symptomatology. Most individuals with sports-related concussion will not score below 15 on the Glasgow Coma Scale, but will present with rapid onset of short-lived neurological impairment, demonstrating no structural changes on traditional magnetic resonance imaging (MRI) and computed tomography (CT) scans. The return-to-play decision is one of the most difficult responsibilities facing the physician, and so far this decision has been primarily based on neurological examination, symptom checklists, and neuropsychological (NP) testing. Diffusion tensor imaging (DTI) may be a more objective tool to assess the severity and recovery of function after concussion. We assessed white matter (WM) fiber tract integrity in varsity level college athletes with sports-related concussion without loss of consciousness, who experienced protracted symptoms for at least 1 month after injury. Evaluation of fractional anisotropy (FA) and mean diffusivity (MD) of the WM skeleton using tract-based spatial statistics (TBSS) revealed a large cluster of significantly increased MD for concussed subjects in several WM fiber tracts in the left hemisphere, including parts of the inferior/superior longitudinal and fronto-occipital fasciculi, the retrolenticular part of the internal capsule, and posterior thalamic and acoustic radiations. Qualitative comparison of average FA and MD suggests that with increasing level of injury severity (ranging from sports-related concussion to severe traumatic brain injury), MD might be more sensitive at detecting mild injury, whereas FA captures more severe injuries. In conclusion, the TBSS analysis used to evaluate diffuse axonal injury of the WM skeleton seems sensitive enough to detect structural changes in sports-related concussion.


Screening ventilation strategies for confined-space manure storage facilities.
Fatalities associated with entry into on-farm confined-space manure storage facilities occur each year. The fatalities are due to asphyxiation or poisoning by exposure to high concentrations of hydrogen sulfide, methane, and carbon dioxide. Forced ventilation has been shown to be an effective way to reduce concentrations of noxious gases to levels that are safe for human entry into these storage facilities. Hydrogen sulfide (H2S) was used as an indicator gas to investigate the effectiveness of forced-air ventilation strategies for eliminating the toxic and oxygen-deficient atmosphere in confined-space manure storage facilities. This article focuses on experimental methods for identifying ventilation strategies that effectively reduce toxic gas (i.e., H2S) concentrations in a fan-ventilated confined-space manure tank to the OSHA permissible exposure limit (PEL) (H2S PEL = 10 ppm) and to 25% of the initial gas concentration. Typical H2S concentration reduction curves during forced-air ventilation were identified in the tank as well. Based on the experimental tests conducted in this research, the most promising candidate ventilation strategies were identified for this rectangular confined-space manure tank with solid, fully slotted, and partially slotted covers. In addition, based on the results of experimental tests, a field-based database was developed for future validation of computational fluid dynamics modeling protocols.


Draft genome sequence of the extremely halophilic Halorubrum sp. SAH-A6 isolated from rock salts of the Danakil depression, Ethiopia.
The draft genome sequence of Halorubrum sp. SAH-A6, isolated from commercial rock salts of the Danakil depression, Ethiopia. The genome comprised 3,325,770 bp, with the G + C content of 68.0%. The strain has many genes which are responsible for secondary metabolites biosynthesis, transport and catabolism as compared to other Halorubrum archaea members. Abundant genes responsible for numerous transport systems, solute accumulation, and aromatic/sulfur decomposition were detected. The first genomic analysis encourages further research on comparative genomics, and biotechnological applications. The NCBI accession number for this genome is SAMN04278861 and ID: 4278861 and strain deposited with accession number KCTC 43215.


Radiation-induced alterations of osteogenic and chondrogenic differentiation of human mesenchymal stem cells.
While human mesenchymal stem cells (hMSCs), either in the bone marrow or in tumour microenvironment could be targeted by radiotherapy, their response is poorly understood. The oxic effects on radiosensitivity, cell cycle progression are largely unknown, and the radiation effects on hMSCs differentiation capacities remained unexplored. Here we analysed hMSCs viability and cell cycle progression in 21% O2 and 3% O2 conditions after medical X-rays irradiation. Differentiation towards osteogenesis and chondrogenesis after irradiation was evaluated through an analysis of differentiation specific genes. Finally, a 3D culture model in hypoxia was used to evaluate chondrogenesis in conditions mimicking the natural hMSCs microenvironment. The hMSCs radiosensitivity was not affected by O2 tension. A decreased number of cells in S phase and an increase in G2/M were observed in both O2 tensions after 16 hours but hMSCs released from the G2/M arrest and proliferated at day 7. Osteogenesis was increased after irradiation with an enhancement of mRNA expression of specific osteogenic genes (alkaline phosphatase, osteopontin). Osteoblastic differentiation was altered since matrix deposition was impaired with a decreased expression of collagen I, probably through an increase of its degradation by MMP-3. After induction in monolayers, chondrogenesis was altered after irradiation with an increase in COL1A1 and a decrease in both SOX9 and ACAN mRNA expression. After induction in a 3D culture in hypoxia, chondrogenesis was altered after irradiation with a decrease in COL2A1, ACAN and SOX9 mRNA amounts associated with a RUNX2 increase. Together with collagens I and II proteins decrease, associated to a MMP-13 expression increase, these data show a radiation-induced impairment of chondrogenesis. Finally, a radiation-induced impairment of both osteogenesis and chondrogenesis was characterised by a matrix composition alteration, through inhibition of synthesis and/or increased degradation. Alteration of osteogenesis and chondrogenesis in hMSCs could potentially explain bone/joints defects observed after radiotherapy.


Acoustic Perturbation Measures Improve with Increasing Vocal Intensity in Individuals With and Without Voice Disorders.
In vocally healthy children and adults, speaking voice loudness differences can significantly confound acoustic perturbation measurements. This study examines the effects of voice sound pressure level (SPL) on jitter, shimmer, and harmonics-to-noise ratio (HNR) in adults with voice disorders and a control group with normal vocal status. This is a matched case-control study. We assessed 58 adult female voice patients matched according to approximate age and occupation with 58 vocally healthy women. Diagnoses included vocal fold nodules (n = 39, 67.2%), polyps (n = 5, 8.6%), and muscle tension dysphonia (n = 14, 24.1%). All participants sustained the vowel /a/ at soft, comfortable, and loud phonation levels. Acoustic voice SPL, jitter, shimmer, and HNR were computed using Praat. The effects of loudness condition, voice SPL, pathology, differential diagnosis, age, and professional voice use level on acoustic perturbation measures were assessed using linear mixed models and Wilcoxon signed rank tests. In both patient and normative control groups, increasing voice SPL correlated significantly (P < 0.001) with decreased jitter and shimmer, and increased HNR. Voice pathology and differential diagnosis were not linked to systematically higher jitter and shimmer. HNR levels, however, were statistically higher in the patient group than in the control group at comfortable phonation levels. Professional voice use level had a significant effect (P < 0.05) on jitter, shimmer, and HNR. The clinical value of acoustic jitter, shimmer, and HNR may be limited if speaking voice SPL and professional voice use level effects are not controlled for. Future studies are warranted to investigate whether perturbation measures are useful clinical outcome metrics when controlling for these effects.


Carbon dynamics and land-use choices: building a regional-scale multidisciplinary model.
Policy enabling tropical forests to approach their potential contribution to global-climate-change mitigation requires forecasts of land use and carbon storage on a large scale over long periods. In this paper, we present an integrated modeling methodology that addresses these needs. We model the dynamics of the human land-use system and of C pools contained in each ecosystem, as well as their interactions. The model is national scale, and is currently applied in a preliminary way to Costa Rica using data spanning a period of over 50 years. It combines an ecological process model, parameterized using field and other data, with an economic model, estimated using historical data to ensure a close link to actual behavior. These two models are linked so that ecological conditions affect land-use choices and vice versa. The integrated model predicts land use and its consequences for C storage for policy scenarios. These predictions can be used to create baselines, reward sequestration, and estimate the value in both environmental and economic terms of including C sequestration in tropical forests as part of the efforts to mitigate global climate change. The model can also be used to assess the benefits from costly activities to increase accuracy and thus reduce errors and their societal costs.


The strategic targeting of females by transnational tobacco companies in South Korea following trade liberalization.
In 1988 South Korea opened its cigarette market to foreign companies under the threat of US trade sanctions. Despite strong social stigma against female smoking in South Korea, and restrictions on tobacco marketing to women and children, smoking rates among young Korean females increased from 1.6% in 1988 to 13% in 1998. Previous analyses describe how Asian countries have been targeted by transnational tobacco companies for new markets, with Asian females offering substantial future growth potential. An understanding of the strategies used by TTCs to increase smoking among Korean females is critical to public health efforts to adopt a stronger gender perspective in implementing the Framework Convention on Tobacco Control. Internal documents of transnational tobacco corporations were systematically searched using keywords focused on the targeting of the female market since market liberalization in 1988. Industry documents were analysed alongside primary and secondary data on the tobacco industry in South Korea. TTCs have targeted Korean females since the late 1980s, conducting market research to understand consumer preferences, cultural characteristics and social changes affecting women and girls. Brands designed to appeal to females have focused on "slim" and "superslim" cigarettes, "light" and "mild" claims, and marketing which appeals to the growing numbers of young women entering the labour force. Strategies for overcoming legal restrictions on marketing to women and children have included the use of company rather than brand names, retail distribution at venues frequented by females, trademark diversification and sponsorship. Given the high male smoking rates in South Korea, tobacco control efforts have given limited attention to girls and women. The limited data available on female smoking behaviour suggests that, despite legal restrictions and social stigma, smoking among females has increased since market opening, notably within younger age groups. In addition to more detailed trend data, there is an urgent need for the development and implementation of gender-sensitive tobacco control measures. Part of South Korea's accession to the FCTC should include emphasis on measures to address the strategic targeting of Korean females by TTCs.


Wing development genes of the pea aphid and differential gene expression between winged and unwinged morphs.
Little is known about when, how or even if the wing development gene network elucidated in Drosophila is deployed in direct-developing insects. Here we identify the wing development genes (as determined in Drosophila) of the pea aphid (Acyrthosiphon pisum), which produces winged or unwinged adults in response to environmental cues. We find that the principal wing development genes studied in Drosophila are present in the aphid genome and that apterous and decapentaplegic exhibit duplications. We followed expression levels of 11 of these developmental genes at embryogenesis and across the nymphal instars. Six showed significant stage-specific expression level effects and apterous1 exhibited significantly different expression levels between winged and unwinged morphs, suggesting this gene acts proximately to realize polyphenic development.


Separation and Identification of Tocotrienol Isomers by HPLC-MS and HPLC-NMR Coupling.
A crude palm-oil extract rich in vitamin E homologues was investigated by HPLC-MS and HPLC-NMR coupling. For mass spectrometry a newly introduced ionization technique called Coordination Ion Spray (CIS) was used. Through the addition of silver ions to the HPLC eluent, the ionization process of nonpolar substances is facilitated. Chromatography and all coupling experiments were conducted on a C(30) column which exhibited an extraordinary shape selectivity and overwhelming sample-loading capability. Experiments were performed with pure methanol as an eluent which proved to be ideal for NMR spectroscopy as well as mass spectrometry. All necessary information for unambiguous structural assignment was collected within 45 min of the LC-NMR experiment and 15 min of the LC-MS experiment. Six compounds were identified, i.e., α-, β-, γ-, and δ-tocotrienol, α-tocoenol, and α-tocopherol.


Treatment of Burkitt lymphoma in equatorial Africa using a simple three-drug combination followed by a salvage regimen for patients with persistent or recurrent disease.
Prior to the introduction of the International Network for Cancer Treatment and Research (INCTR) protocol INCTR 03-06, survival of patients with Burkitt lymphoma at four tertiary care centres in equatorial Africa was probably no more than 10-20%. The results reported here for 356 patients have demonstrated marked improvement in survival through the use of a uniform treatment protocol consisting of cyclophosphamide, methotrexate, vincristine, and intrathecal therapy, and the introduction of non-cross resistant second-line (salvage) therapy, consisting of ifosfamide, mesna, etoposide and cytarabine, when patients failed to achieve a complete response to first-line therapy or relapsed early. Overall survival rates of 67% and 62% were observed at 1 and 2 years (relapse is rare after 1 year of remission). Of interest was the small impact of cerebrospinal fluid (CSF) and bone marrow involvement on outcome. However, the presence or absence of abdominal involvement clearly defined two prognostic groups. An additional finding was the association between CSF pleocytosis and orbital tumours, suggesting that spread of tumour cells to the central nervous system may sometimes occur via direct involvement of cranial nerves in the orbit. Survival rates may be increased in patients with abdominal involvement by combining first- and second-line therapy, but verification will require a further clinical study.


Effect of Interleukin-29 on Interferon-α Secretion by Peripheral Blood Mononuclear Cells.
The effect of interleukin (IL)-29, a new therapeutic agent similar to type I interferons (IFNs), on IFN-α secretion of human plasmacytoid dendritic cells (pDCs) has not been studied. Therefore, in this study, we aimed to clarify the effect of IL-29 on IFN-α secretion of pDCs using human peripheral blood mononuclear cells (PBMCs) in the presence of cytosine-phosphate-guanosinemotif-containing oligodeoxy nucleotides (CpG). In this experimental and prospective study, PBMCs were ob- tained from 11 healthy volunteers and divided into four culture conditions: I. control, II. CpG treatment, III. IL-29 treatment and IV. CpG plus IL-29 treatment. The amount of IFN-α secretion was measured from each culture supernatant by flow cytometry using the flowcytomix apparatus (eBioscience, Vienna, Austria). Fractional IFN-α production of the cultured PBMCs was measured by intracellular staining using the cytomics FC 500 system (Beckman Coulter, Brea, CA, USA) with CXP Software. The mean ± standard deviation (SD) of supernatant IFN-α secretion per pDC/μL was 5.7 ± 9.3 pg/mL/count/µL for condition I, 1071.5 ± 1026.6 pg/mL/count/µL for condition II, 14.1 ± 21.1 pg/mL/count/µL for condition III, and 1913.9 ± 1525.9 pg/mL/count/µL for condition IV. There were statistically significant differences between conditions I and II as well as betweenconditions II and IV. Intracellular IFN-α production was only detectable in the pDC fraction from one culture; the production amount was similar between the cells treated with CpG and those treated with CpG plus IL-29. Natural killer (NK) cell production of IFN-α was observed in two out of three cultures and one culture showed IFN- α production in the monocyte fraction. IL-29 alone did not show any effect on IFN-α secretion of PBMCs. However, the addition of CpG along with IL-29 enhanced IFN-α secretion of PBMCs. Given that pDCs are the major secretors of IFN-α in peripheral blood, this result has suggested the possibility that IL-29 has an enhancing effect in human pDC IFN-α secretion. Although the supernatant IFN-α secretion was not directly correlated with pDCs's intracellular IFN-α production in this study, prolonged incubation of pDC and other PB subsets with CpG or IL-29 for over 4 hours could be applied in future studies. These studies would help to elucidate the mechanism of action of IL-29 in human pDCs associated with viral infections.


Perfect selective metamaterial solar absorbers.
In this work, we numerically investigate the radiative properties of metamaterial nanostructures made of two-dimensional tungsten gratings on a thin dielectric spacer and an opaque tungsten film from UV to mid-infrared region as potential selective solar absorbers. The metamaterial absorber with single-sized tungsten patches exhibits high absorptance in the visible and near-infrared region due to several mechanisms such as surface plasmon polaritons, magnetic polaritons, and intrinsic bandgap absorption of tungsten. Geometric effects on the resonance wavelengths and the absorptance spectra are studied, and the physical mechanisms are elucidated in detail. The absorptance could be further enhanced in a broader spectral range with double-sized metamaterial absorbers. The total solar absorptance of the optimized metamaterial absorbers at normal incidence could be more than 88%, while the total emittance is less than 3% at 100°C, resulting in total photon-to-heat conversion efficiency of 86% without any optical concentration. Moreover, the metamaterial solar absorbers exhibit quasi-diffuse behaviors as well as polarization independence. The results here will facilitate the design of novel highly efficient solar absorbers to enhance the performance of various solar energy conversion systems.


Legacy of Multiple Stressors: Responses of Gastropod Larvae and Juveniles to Ocean Acidification and Nutrition.
Ocean acidification poses a significant threat to calcifying invertebrates by negatively influencing shell deposition and growth. An organism's performance under ocean acidification is not determined by the susceptibility of one single life-history stage, nor is it solely controlled by the direct physical consequences of ocean acidification. Shell development by one life-history stage is sometimes a function of the pH or pCO2 levels experienced during earlier developmental stages. Furthermore, environmental factors such as access to nutrition can buffer organismal responses of calcifying invertebrates to ocean acidification, or they can function as a co-occurring stressor when access is low. We reared larvae and juveniles of the planktotrophic marine gastropod Crepidula fornicata through combined treatments of nutritional stress and low pH, and we monitored how multiple stressors endured during the larval stage affected juvenile performance. Shell growth responded non-linearly to decreasing pH, significantly declining between pH 7.6 and pH 7.5 in larvae and juveniles. Larval rearing at pH 7.5 reduced juvenile growth as a carryover effect. Larval rearing at pH 7.6 reduced subsequent juvenile growth despite the absence of a negative impact on larval growth, demonstrating a latent effect. Low larval pH magnified the impact of larval nutritional stress on competence for metamorphosis and increased carryover effects of larval nutrition on juvenile growth. Trans-life-cycle effects of larval nutrition were thus modulated by larval exposure to ocean acidification.


Delivering Peer-Based Support in Prisons During the COVID Pandemic and Lockdown: Innovative Activities Delivered by People Who Care.
During COVID and lockdown, many prisoners have not only been affected by infection transmission in crowded and ill-equipped institutions, they have also been separated from a range of supports, including loss of family and professional supports and support for prisoners with addiction and/or mental health problems has been disrupted. This paper reports on evidence of how peer-based recovery organizations have attempted to mitigate these adverse effects, based on a case study of one prison in the North-West of England, using a range of routine reporting data and original research data. The paper shows how prison-based peer recovery support has not only continued through lockdown but grown both in the prison and in continuing care on release. The key conclusion is that Lived Experience Recovery Organizations (LEROs) have a vital role to play in offering continuing care to prison populations both to support early recovery and to sustain change around release back into the community, in COVID but also more generally.


Glycolysis in contracting rat skeletal muscle is controlled by factors related to energy state.
The control of glycolysis in contracting muscle is not fully understood. The aim of the present study was to examine whether activation of glycolysis is mediated by factors related to the energy state or by a direct effect of Ca2+ on the regulating enzymes. Extensor digitorum longus muscles from rat were isolated, treated with cyanide to inhibit aerobic ATP production and stimulated (0.2 s trains every 4 s) until force was reduced to 70% of initial force (control muscle, referred to as Con). Muscles treated with BTS (N-benzyl-p-toluene sulfonamide), an inhibitor of cross-bridge cycling without affecting Ca2+ transients, were stimulated for an equal time period as Con. Energy utilization by the contractile apparatus (estimated from the observed relation between ATP utilization and force-time integral) was 60% of total. In BTS, the force-time integral and ATP utilization were only 38 and 58% of those in Con respectively. Glycolytic rate in BTS was only 51% of that in Con but the relative contribution of ATP derived from PCr (phosphocreatine) and glycolysis and the relation between muscle contents of PCr and Lac (lactate) were not different. Prolonged cyanide incubation of quiescent muscle (low Ca2+) did not change the relation between PCr and Lac. The reduced glycolytic rate in BTS despite maintained Ca2+ transients, and the unchanged PCr/Lac relation in the absence of Ca2+ transients, demonstrates that Ca2+ is not the main trigger of glycogenolysis. Instead the preserved relative contribution of energy delivered from PCr and glycolysis during both conditions suggests that the glycolytic rate is controlled by factors related to energy state.


Multiple impacts of the plant growth-promoting rhizobacterium Variovorax paradoxus 5C-2 on nutrient and ABA relations of Pisum sativum.
Resolving the physiological mechanisms by which rhizobacteria enhance plant growth is difficult, since many such bacteria contain multiple plant growth-promoting properties. To understand further how the 1-aminocyclopropane-1-carboxylate (ACC) deaminase (ACCd)-containing rhizobacterium Variovorax paradoxus 5C-2 affects plant growth, the flows and partitioning of mineral nutrients and abscisic acid (ABA) and ABA metabolism were studied in pea (Pisum sativum) plants following rhizosphere bacterial inoculation. Although root architecture was not affected, inoculation increased root and shoot biomass, and stomatal conductance, by 20, 15, and 24%, respectively, and increased N, P, K, Ca, and Mg uptake by 16, 81, 50, 46, and 58%, respectively. P deposition in inoculated plant roots was 4.9 times higher than that in uninoculated controls. Rhizobacterial inoculation increased root to shoot xylem flows and shoot to root phloem flows of K by 1.8- and 2.1-fold, respectively. In control plants, major sinks for K deposition were the roots and upper shoot (43% and 49% of total uptake, respectively), while rhizobacterial inoculation increased K distribution to the lower shoot at the expense of other compartments (xylem, phloem, and upper shoot). Despite being unable to metabolize ABA in vitro, V. paradoxus 5C-2 decreased root ABA concentrations and accumulation by 40-60%. Although inoculation decreased xylem ABA flows, phloem ABA flows increased. Whether bacterial ACCd attenuates root to shoot ABA signalling requires further investigation, since ABA is critical to maintain growth of droughted plants, and ACCd-containing organisms have been advocated as a means of minimizing growth inhibition of plants in drying soil.


Pulsed Azidohomoalanine Labeling in Mammals (PALM) Detects Changes in Liver-Specific LKB1 Knockout Mice.
Quantification of proteomes by mass spectrometry has proven to be useful to study human pathology recapitulated in cellular or animal models of disease. Enriching and quantifying newly synthesized proteins (NSPs) at set time points by mass spectrometry has the potential to identify important early regulatory or expression changes associated with disease states or perturbations. NSP can be enriched from proteomes by employing pulsed introduction of the noncanonical amino acid, azidohomoalanine (AHA). We demonstrate that pulsed introduction of AHA in the feed of mice can label and identify NSP from multiple tissues. Furthermore, we quantitate differences in new protein expression resulting from CRE-LOX initiated knockout of LKB1 in mouse livers. Overall, the PALM strategy allows for the first time in vivo labeling of mouse tissues to differentiate protein synthesis rates at discrete time points.


Haplotype estimation using sequencing reads.
High-throughput sequencing technologies produce short sequence reads that can contain phase information if they span two or more heterozygote genotypes. This information is not routinely used by current methods that infer haplotypes from genotype data. We have extended the SHAPEIT2 method to use phase-informative sequencing reads to improve phasing accuracy. Our model incorporates the read information in a probabilistic model through base quality scores within each read. The method is primarily designed for high-coverage sequence data or data sets that already have genotypes called. One important application is phasing of single samples sequenced at high coverage for use in medical sequencing and studies of rare diseases. Our method can also use existing panels of reference haplotypes. We tested the method by using a mother-father-child trio sequenced at high-coverage by Illumina together with the low-coverage sequence data from the 1000 Genomes Project (1000GP). We found that use of phase-informative reads increases the mean distance between switch errors by 22% from 274.4 kb to 328.6 kb. We also used male chromosome X haplotypes from the 1000GP samples to simulate sequencing reads with varying insert size, read length, and base error rate. When using short 100 bp paired-end reads, we found that using mixtures of insert sizes produced the best results. When using longer reads with high error rates (5-20 kb read with 4%-15% error per base), phasing performance was substantially improved.


Clinicopathological characteristics and outcome of 31 patients with ETV6-NTRK3 fusion gene confirmed (mammary analogue) secretory carcinoma of salivary glands.
In 2010, a new subtype of salivary gland cancer (SGC), (mammary analogue) secretory carcinoma (SC), was defined, characterized by the ETV6-NTRK3 fusion gene. As clinical behavior and outcome data of this histological subtype tumor are still sparse, we aimed to describe the clinicopathological course and outcome of a series of translocation positive SC patients. We re-evaluated the pathological diagnosis of a subset of SGCs, diagnosed in 4 of 8 Dutch head and neck centers. Subsequently, tumors with a morphological resemblance to SC were tested for the ETV6-NTRK3 fusion gene using RT-PCR. Furthermore, patients prospectively diagnosed with SC were included. The clinical characteristics and outcomes were retrieved from the patient files. Thirty-one patients with ETV6-NTRK3 fusion gene positive SC were included. The median age was 49 years, 17 patients (55%) were male. Eighteen tumors (58%) arose in the parotid gland. One patient presented with lymph node metastasis. All patients underwent tumor resection and 4 patients had a neck dissection. Four patients had re-resection and 15 patients (48%) received postoperative radiotherapy. One patient developed a local recurrence, no regional recurrences or distant metastases were observed. After a median follow-up of 49 months the 5- and 10-year overall survival were 95%, the 5- and 10-year disease free survival were 89%. The clinical course of SC is favorable with a low rate of locoregional recurrence and excellent survival. Given the low incidence of nodal metastases, elective neck treatment, i.e. surgery and/or radiotherapy, does not seem to be indicated.


Additive effect of neutralizing antibody and antiviral drug treatment in preventing virus escape and persistence.
Poorly cytopathic or noncytopathic viruses can escape immune surveillance and establish a chronic infection. Here we exploited the strategy of combining antiviral drug treatment with the induction of a neutralizing antibody response to avoid the appearance of neutralization-resistant virus variants. Despite the fact that H25 immunoglobulin transgenic mice infected with lymphocytic choriomeningitis virus mounted an early neutralizing antibody response, the virus escaped from neutralization and persisted. After ribavirin treatment of H25 transgenic mice, the appearance of neutralization-resistant virus was prevented and virus was cleared. Thus, the combination of virus-neutralizing antibodies and chemotherapy efficiently controlled the infection, whereas each defense line alone did not. Similar additive effects may be unexpectedly efficient and beneficial in humans after infections with persistent viruses such as hepatitis C virus and hepatitis B virus and possibly human immunodeficiency virus.


Organization of an actin filament-membrane complex. Filament polarity and membrane attachment in the microvilli of intestinal epithelial cells.
The association of actin filaments with membranes is now recognized as an important parameter in the motility of nonmuscle cells. We have investigated the organization of one of the most extensive and highly ordered actin filament-membrane complexes in nature, the brush border of intestinal epithelial cells. Through the analysis of isolated, demembranated brush borders decorated with the myosin subfragment, S1, we have determined that all the microvillar actin filaments have the same polarity. The S1 arrowhead complexes point away from the site of attachment of actin filaments at the apical tip of the microvillar membrane. In addition to the end-on attachment of actin filaments at the tip of the microvillus, these filaments are also connected to the plasma membrane all along their lengths by periodic (33 nm) cross bridges. These bridges were best observed in isolated brush borders incubated in high concentrations of Mg++. Their visibility is attributed to the induction of actin paracrystals in the filament bundles of the microvilli. Finally, we present evidence for the presence of myosinlike filaments in the terminal web region of the brush border. A model for the functional organization of actin and myosin in the brush border is presented.