Canonical and noncanonical Wnt signaling in neural stem/progenitor cells.
The first mammalian Wnt to be discovered, Wnt-1, was found to be essential for the development of a large part of the mouse brain over 25 years ago. We have since learned that Wnt family secreted glycolipoproteins, of which there are nineteen, which activate a diverse network of signals that are particularly important during embryonic development and tissue regeneration. Wnt signals in the developing and adult brain can drive neural stem cell self-renewal, expansion, asymmetric cell division, maturation and differentiation. The molecular events taking place after a Wnt binds to its cell-surface receptors are complex and, at times, controversial. A deeper understanding of these events is anticipated to lead to improvements in the treatment of neurodegenerative diseases and stem cell-based replacement therapies. Here, we review the roles played by Wnts in neural stem cells in the developing mouse brain, at neurogenic sites of the adult mouse and in neural stem cell culture models.


Packaging of pharmaceuticals: still too many dangers but several encouraging initiatives.
(1) In 2006 in France, several drugs sold in poorly designed packaging exposed patients to a risk of serious adverse effects. (2) In 2006, Prescrire used a standardised methodology to examine the packaging of all new pharmaceutical products (656 different boxes) assessed in the New Products section of our French edition, la revue Prescrire. About 75% of these boxes contained tablets or capsules, mostly in blister packs. (3) Poor labelling remains a major problem. The international nonproprietary names (INN) is hard to spot on most boxes of patented brand-name drugs and is often overshadowed by the brand name. The primary packaging of many products does not even include the INN. (4) Two particularly ambiguous types of labelling are becoming more common on blister packs: pre-cut multiple-unit blister packs on which the labelling is truncated when a unit blister is removed; and blister packs on which the labelling spans two blisters, creating a risk of overdose. (5) The use of colours is frequently inappropriate. In particular, irrelevant information is often highlighted unnecessarily, while other, important information is barely visible. (6) Too many devices for oral administration create a risk of misuse. Very few are graduated in units of weight. Most are graduated in millilitres, obliging caregivers to use conversion charts and thus creating a risk of dosing errors. Devices graduated in kg bodyweight can also lead to dosing errors. (7) The labelling of some injectable drugs is barely legible. The various models of plastic ampoules, that are gradually replacing glass ampoules, can represent a danger because they resemble other plastic ampoules containing products administered by different routes. Packaging that does not provide a syringe or needle can cause problems for caregivers and represents another potential source of error. (8) Many of the patient information leaflets examined in 2006 had the same flaws as previously observed, i.e. uneven information quality, discrepancies between different sections, and out-of-date information. More and more French leaflets now include insets offering "Health Advice". There are better and worse examples, but there is no guarantee that they have been properly reviewed by the regulatory agency. (9) Increasingly drug boxes include pictograms, even though several studies have shown they are often difficult to interpret. And most boxes of generics also now include standard dosing schedules that are not always appropriate and may create a risk of dosing errors with potentially serious consequences. (10) 67 multidose bottles examined in 2006 had no childproof safety cap. Some contained psychotropics, which can have life-threatening effects if accidentally consumed in large amounts. (11) Some manufacturers have adopted realistic solutions to these problems. In particular, generics manufacturers again improved product labelling in 2006 (emphasis on the INN), appropriate use of colours for dose differentiation, and, encouragingly, far more Braille labels. (12) In 2006, the French regulatory agency introduced several measures aimed at improving the labelling of ampoules containing some injectable drugs. The impact of these measures was visible on several products marketed in 2006, including plastic vials of solutions for nebulization. (13) Several other examples of well-designed packaging were seen: safety devices on prefilled syringes; a childproof safety device; a tamperproof ring; unit-dose blister packs; clearly written patient leaflets; and the use of clear and appropriate symbols and pictograms. (14) In practice, in view of the large number of incidents recorded in 2006, and the plethora of packaging designs, caregivers should take time to analyse and discuss drug packaging. In this way, they will be in a position to distinguish between good and bad drug packaging, and to anticipate the risks associated with poorly designed packaging. (15) There are many ways in which drug regulatory authorities can help to ensure that drugs are sold in safe packaging. The French regulatory agency's work on the labelling of injectable drugs is an encouraging step. European Directive 2004/27/EC on medicines for human use provides for improvements in labelling (e.g. Braille) and patient information leaflets. Transposition of these measures into French law should lead to a number of improvements, provided the relevant regulations and guidelines place patients' interests first.


Epicardial adipose tissue thickness in patients with ankylosing spondylitis.
The purpose of our study was to measure epicardial adipose tissue (EAT) thickness as a novel indicator of atherosclerosis and cardiovascular risk factor in ankylosing spondylitis (AS) patients and to show the relationship with clinical parameters and inflammatory markers. Forty AS patients (42.75 ± 12.43 years) and 40 healthy individuals with no cardiovascular risk factor as the control group (43.02 ± 14.78 years) were included in the study. Carotid intima-media thickness (CIMT) and EAT thickness were measured in AS patients and the control group. Total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, glucose, erythrocyte sedimentation rate, urea, and blood pressure were investigated in both groups. In addition, the Bath Ankylosing Spondylitis Functional Index (BASFI) and the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) were used to evaluate the association between clinical findings and CIMT and EAT in the patient group. CIMT and EAT thickness were higher in the AS patients compared to the control group. CIMT was 0.76 ± 0.19 and 0.57 ± 0.12 mm (p < 0.001) and EAT thickness was 4.35 ± 1.56 and 3.03 ± 0.94 mm (p < 0.001) in the AS and control groups, respectively. A correlation was determined between EAT thickness and CIMT. Triglyceride level, patient age, blood pressure, and duration of disease were correlated with both CIMT and EAT thickness. Increased CIMT and EAT thickness in AS patients compared to the control group shows a risk for subclinical atherosclerosis and cardiovascular disease.


Inertial effects in diffusion-limited reactions.
Diffusion-limited reactions are commonly found in biochemical processes such as enzyme catalysis, colloid and protein aggregation and binding between different macromolecules in cells. Usually, such reactions are modeled within the Smoluchowski framework by considering purely diffusive boundary problems. However, inertial effects are not always negligible in real biological or physical media on typical observation time frames. This is all the more so for non-bulk phenomena involving physical boundaries, that introduce additional time and space constraints. In this paper, we present and test a novel numerical scheme, based on event-driven Brownian dynamics, that allows us to explore a wide range of velocity relaxation times, from the purely diffusive case to the underdamped regime. We show that our algorithm perfectly reproduces the solution of the Fokker-Planck problem with absorbing boundary conditions in all the regimes considered and is thus a good tool for studying diffusion-guided reactions in complex biological environments.


Episodic ataxia type 1 mutations differentially affect neuronal excitability and transmitter release.
Heterozygous mutations of KCNA1, the gene encoding potassium channel Kv1.1 subunits, cause episodic ataxia type 1 (EA1), which is characterized by paroxysmal cerebellar incoordination and interictal myokymia. Some mutations are also associated with epilepsy. Although Kv1.1-containing potassium channels play important roles in neuronal excitability and neurotransmitter release, it is not known how mutations associated with different clinical features affect the input-output relationships of individual neurons. We transduced rat hippocampal neurons, which were cultured on glial micro-islands, with lentiviruses expressing wild-type or mutant human KCNA1, and injected either depolarizing currents to evoke action potentials or depolarizing voltage commands to evoke autaptic currents. alpha-Dendrotoxin and tetraethylammonium allowed a pharmacological dissection of potassium currents underlying excitability and neurotransmission. Overexpression of wild-type Kv1.1 decreased both neuronal excitability and neurotransmitter release. By contrast, the C-terminus-truncated R417stop mutant, which is associated with severe drug-resistant EA1, had the opposite effect: increased excitability and release probability. Another mutant, T226R, which is associated with EA1 that is complicated by contractures and epilepsy, had no detectable effect on neuronal excitability; however, in common with R417stop, it markedly enhanced neurotransmitter release. The results provide direct evidence that EA1 mutations increase neurotransmitter release, and provide an insight into mechanisms underlying the phenotypic differences that are associated with different mutations.


Biomarkers in HCV-related mixed cryoglobulinemia patients withnon-Hodgkin lymphoma.
Chronic Hepatitis C virus (HCV) infection can cause severe extrahepatic manifestations, such as mixed cryoglobulins (MC), up to the development of B cell nonHodgkin's lymphoma (B-NHL). Mechanisms transforming of HCV infection into lymphoproliferative and/or autoimmune disorders are still poorly understood. In course of HCV infection, the sustained virus-driven antigenic stimulation may probably induce a B-cell clonal expansion. Measurements of serum free light chains (FLCs) levels, considered as a direct marker of B cell activity, are analyzed with increasing interest in clinical practice, for diagnosis, monitoring and follow-up of plasma cell dyscrasia. Syndecan-1 (CD138) is a transmembrane heparan sulfate proteoglycan expressed and actively shed by most myeloma cells. Membrane CD138 represents the major receptor protein for HCV attachment to the hepatocyte surface and high levels of circulating sCD138 levels are detected in patients at early stage of B-cell chronic lymphocytic leukemia. This study is aimed to evaluate sCD138 and FLC levels as diagnostic biomarkers of HCV-related MC with B-NHL. We enrolled 35 HCV-MC-NHL patients, characterized for the specific type of cryoglobulins, and 25 healthy blood donors (HBD) as negative control. Serum sCD138 levels were determined using ELISA kits specific for human sCD138. Serum FLCs were assessed by means of the turbidimetric assay. We found that serum levels of sCD138, as well as FLCs, were significantly higher in patients than in HBD (p<0.001). In agreement with the definition of HCV-driven lymphoproliferative disorders as the consequence of a multifactorial and multistep pathogenetic process, we suggest that sCD138 and FLCs could be considered putative independent markers of worsening progression of the disease.


Role of mitochondrial ATP-sensitive potassium channel-mediated PKC-ε in delayed protection against myocardial ischemia/reperfusion injury in isolated hearts of sevoflurane-preconditioned rats.
This study aimed to determine the role of mitochondrial adenosine triphosphate-sensitive potassium (mitoKATP) channels and protein kinase C (PKC)-ε in the delayed protective effects of sevoflurane preconditioning using Langendorff isolated heart perfusion models. Fifty-four isolated perfused rat hearts were randomly divided into 6 groups (n=9). The rats were exposed for 60 min to 2.5% sevoflurane (the second window of protection group, SWOP group) or 33% oxygen inhalation (I/R group) 24 h before coronary occlusion. The control group (CON) and the sevoflurane group (SEVO) group were exposed to 33% oxygen and 2.5% sevoflurane for 60 min, respectively, without coronary occlusion. The mitoKATP channel inhibitor 5-hydroxydecanoate (5-HD) was given 30 min before sevoflurane preconditioning (5-HD+SWOP group). Cardiac function indices, infarct sizes, serum cardiac troponin I (cTnI) concentrations, and the expression levels of phosphorylated PKC-ε (p-PKC-ε) and caspase-8 were measured. Cardiac function was unchanged, p-PKC-ε expression was upregulated, caspase-8 expression was downregulated, cTnI concentrations were decreased, and the infarcts were significantly smaller (P<0.05) in the SWOP group compared with the I/R group. Cardiac function was worse, p-PKC-ε expression was downregulated, caspase-8 expression was upregulated, cTnI concentration was increased and infarcts were larger in the 5-HD+SWOP group (P<0.05) compared with the SWOP group. The results suggest that mitoKATP channels are involved in the myocardial protective effects of sevoflurane in preconditioning against I/R injury, by regulating PKC-ε phosphorylation before ischemia, and by downregulating caspase-8 during reperfusion.


Shockable rhythms and defibrillation during in-hospital pediatric cardiac arrest.
To analyze the results of cardiopulmonary resuscitation (CPR) that included defibrillation during in-hospital cardiac arrest (IH-CA) in children. A prospective multicenter, international, observational study on pediatric IH-CA in 12 European and Latin American countries, during 24 months. Data from 502 children between 1 month and 18 years were collected using the Utstein template. Patients with a shockable rhythm that was treated by electric shock(s) were included. The primary endpoint was survival at hospital discharge. Univariate logistic regression analysis was performed to find outcome factors. Forty events in 37 children (mean age 48 months, IQR: 7-15 months) were analyzed. An underlying disease was present in 81.1% of cases and 24.3% had a previous CA. The main cause of arrest was a cardiac disease (56.8%). In 17 episodes (42.5%) ventricular fibrillation (VF) or pulseless ventricular tachycardia (pVT) was the first documented rhythm, and in 23 (57.5%) it developed during CPR efforts. In 11 patients (27.5%) three or more shocks were needed to achieve defibrillation. Return of spontaneous circulation (ROSC) was obtained in 25 cases (62.5%), that was sustained in 20 (50.0%); however only 12 children (32.4%) survived to hospital discharge. Children with VF/pVT as first documented rhythm had better sustained ROSC (64.7% vs. 39.1%, p=0.046) and survival to hospital discharge rates (58.8% vs. 21.7%, p=0.02) than those with subsequent VF/pVT. Survival rate was inversely related to duration of CPR. Clinical outcome was not related to the cause or location of arrest, type of defibrillator and waveform, energy dose per shock, number of shocks, or cumulative energy dose, although there was a trend to better survival with higher doses per shock (25.0% with <2Jkg(-1), 43.4% with 2-4Jkg(-1) and 50.0% with >4Jkg(-1)) and worse with higher number of shocks and cumulative energy dose. The termination of pediatric VF/pVT in the IH-CA setting is achieved in a low percentage of instances with one electrical shock at 4Jkg(-1). When VF/pVT is the first documented rhythm, the results of defibrillation are better than in the case of subsequent VF/pVT. No clear relationship between defibrillation protocol and ROSC or survival has been observed. The optimal pediatric defibrillation dose remains to be determined; therefore current resuscitation guidelines cannot be considered evidence-based, and additional research is needed.


Asking Youth Questions About Suicide Risk in the Pediatric Emergency Department: Results From a Qualitative Analysis of Patient Opinions.
The emergency department (ED) is a promising setting to screen youth for suicide risk. Patient reactions to questions about suicidal thoughts and behaviors during their ED visit have implications for how screening is introduced, developed, and implemented. The current study is a qualitative investigation into patient opinions about screening for suicide risk in the pediatric ED. As part of a subset of a multisite study, 165 participants, 10 to 21 years old, were included in this sub-analysis. Ninety percent (148/165) of participants supported suicide risk screening. Reasons youth support screening included prevention of suicide, detection of at-risk youth, and a lack of other social support. Overall, pediatric patients agreed with suicide risk screening in the ED, citing similar reasons as in a previous investigation, further demonstrating acceptability of suicide risk screening in this setting. A small subset of youth (10%; 17/165) did not support screening for reasons that included a desire to focus on their chief (i.e., nonpsychiatric) presenting concern and fear of iatrogenic risk. Understanding patient opinions, including those in support of and in opposition to screening, can inform implementation practices. Further education about the importance of suicide risk assessment may be a helpful first step in instituting universal screening efforts.


Mapping and validation of quantitative trait loci associated with concentrations of 16 elements in unmilled rice grain.
QTLs controlling the concentrations elements in rice grain were identified in two mapping populations. The QTLs were clustered such that most genomic regions were associated with more than one element. In this study, quantitative trait loci (QTLs) affecting the concentrations of 16 elements in whole, unmilled rice (Oryza sativa L.) grain were identified. Two rice mapping populations, the 'Lemont' × 'TeQing' recombinant inbred lines (LT-RILs), and the TeQing-into-Lemont backcross introgression lines (TILs) were used. To increase opportunity to detect and characterize QTLs, the TILs were grown under two contrasting field conditions, flooded and irrigated-but-unflooded. Correlations between the individual elements and between each element with grain shape, plant height, and time of heading were also studied. Transgressive segregation was observed among the LT-RILs for all elements. The 134 QTLs identified as associated with the grain concentrations of individual elements were found clustered into 39 genomic regions, 34 of which were found associated with grain element concentration in more than one population and/or flooding treatment. More QTLs were found significant among flooded TILs (92) than among unflooded TILs (47) or among flooded LT-RILs (40). Twenty-seven of the 40 QTLs identified among the LT-RILs were associated with the same element among the TILs. At least one QTL per element was validated in two or more population/environments. Nearly all of the grain element loci were linked to QTLs affecting additional elements, supporting the concept of element networks within plants. Several of the grain element QTLs co-located with QTLs for grain shape, plant height, and days to heading; but did not always differ for grain elemental concentration as predicted by those traits alone. A number of interesting patterns were found, including a strong Mg–P–K complex.


Association of Clinical Reactivity with Sensitization to Allergen Components in Multifood-Allergic Children.
Thirty percent of children with food allergies have multiple simultaneous allergies; however, the features of these multiple allergies are not well characterized serologically or clinically. We comprehensively evaluated 60 multifood-allergic patients by measuring serum IgE to key allergen components, evaluating clinical histories and medication use, performing skin tests, and conducting double-blind, placebo-controlled food challenges (DBPCFCs). Sixty participants with multiple food allergies were characterized by clinical history, DBPCFCs, total IgE, specific IgE, and component-resolved diagnostics (IgE and IgG4) data. The food allergens tested were almond, egg, milk, sesame, peanut, pecan, walnut, hazelnut, cashew, pistachio, soy, and wheat. Our data demonstrate that of the reactions observed during a graded DBPCFC, gastrointestinal reactions occurred more often in boys than in girls, as well as in individuals with high levels of IgE to 2S albumins from cashew, walnut, and hazelnut. Certain food allergies often occurred concomitantly in individuals (ie, cashew/pistachio and walnut/pecan/hazelnut). IgE testing to components further corroborated serological relationships between and among these clustered food allergies. Associations of certain food allergies were shown by DBPCFC outcomes as well as by correlations in IgE reactivity to structurally related food allergen components. Each of these criteria independently demonstrated a significant association between allergies to cashew and pistachio, as well as among allergies to walnut, pecan, and hazelnut.


Mepolizumab effectiveness and identification of super-responders in severe asthma.
Severe asthma is a high-burden disease. Real-world data on mepolizumab in patients with severe eosinophilic asthma is needed to assess whether the data from randomised controlled trials are applicable in a broader population.The Australian Mepolizumab Registry (AMR) was established with an aim to assess the use, effectiveness and safety of mepolizumab for severe eosinophilic asthma in Australia.Patients (n=309) with severe eosinophilic asthma (median age 60 years, 58% female) commenced mepolizumab. They had poor symptom control (median Asthma Control Questionnaire (ACQ)-5 score of 3.4), frequent exacerbations (median three courses of oral corticosteroids (OCS) in the previous 12 months), and 47% required daily OCS. Median baseline peripheral blood eosinophil level was 590 cells·µL-1 Comorbidities were common: allergic rhinitis 63%, gastro-oesophageal reflux disease 52%, obesity 46%, nasal polyps 34%.Mepolizumab treatment reduced exacerbations requiring OCS compared with the previous year (annualised rate ratio 0.34 (95% CI 0.29-0.41); p<0.001) and hospitalisations (rate ratio 0.46 (95% CI 0.33-0.63); p<0.001). Treatment improved symptom control (median ACQ-5 reduced by 2.0 at 6 months), quality of life and lung function. Higher blood eosinophil levels (p=0.003) and later age of asthma onset (p=0.028) predicted a better ACQ-5 response to mepolizumab, whilst being male (p=0.031) or having body mass index ≥30 (p=0.043) predicted a lesser response. Super-responders (upper 25% of ACQ-5 responders, n=61, 24%) had a higher T2 disease burden and fewer comorbidities at baseline.Mepolizumab therapy effectively reduces the significant and long-standing disease burden faced by patients with severe eosinophilic asthma in a real-world setting.


Intraepithelial lymphocyte distribution differs between the bulb and the second part of duodenum.
Evaluation of intraepithelial duodenal lymphocytosis (IDL) is important in celiac disease (CD). There is no established cut-off value for increased number of IELs in the bulb.We therefore investigated the relation between IEL counts in the bulb and duodenal specimens in non-celiac subjects. The number of CD3+ IELs was determined in specimens from the second part of the duodenum and from the bulb in 34 non-celiac subjects. The numbers of IELs in the villus tip and sides were counted and the quotient tip/side was calculated. HLA DQ2/DQ8 and serum antibodies against transglutaminase were analysed. The mean number of IELs per 100 enterocytes (95% CI) in specimens was 14.7 (11.8-17.6) in the bulb, and 21.2 (17.0-25.5) in the second part of the duodenum (p<0.01). There was no difference in IEL count or distribution comparing patients carrying or lacking HLA DQ2/DQ8. IEL count in non-celiac, HLA DQ2/DQ8 positive or negative patients is significantly lower in the bulb than in the second part of the duodenum. These findings implicate that the site of biopsy should be taken into account when considering duodenal lymphocytosis.


Examining inequalities in uptake of maternal health care and choice of provider in underserved urban areas of Mumbai, India: a mixed methods study.
Discussions of maternity care in developing countries tend to emphasise service uptake and overlook choice of provider. Understanding how families choose among health providers is essential to addressing inequitable access to care. Our objectives were to quantify the determinants and choice of maternity care provider in Mumbai's informal urban settlements, and to explore the reasons underlying their choices. The study was conducted in informal urban communities in eastern Mumbai. We developed regression models using data from a census of married women aged 15-49 to test for associations between maternal characteristics and uptake of care and choice of provider. We then conducted seven focus group discussions and 16 in-depth interviews with purposively selected participants, and used grounded theory methods to examine the reasons for their choices. Three thousand eight hundred forty-eight women who had given birth in the preceding 2 years were interviewed in the census. The odds of institutional prenatal and delivery care increased with education, economic status, and duration of residence in Mumbai, and decreased with parity. Tertiary public hospitals were the commonest site of care, but there was a preference for private hospitals with increasing socio-economic status. Women were more likely to use tertiary public hospitals for delivery if they had fewer children and were Hindu. The odds of delivery in the private sector increased with maternal education, wealth, age, recent arrival in Mumbai, and Muslim faith. Four processes were identified in choosing a health care provider: exploring the options, defining a sphere of access, negotiating autonomy, and protective reasoning. Women seeking a positive health experience and outcome adopted strategies to select the best or most suitable, accessible provider. In Mumbai's informal settlements, institutional maternity care is the norm, except among recent migrants. Poor perceptions of primary public health facilities often cause residents to bypass them in favour of tertiary hospitals or private sector facilities. Families follow a complex selection process, mediated by their ability to mobilise economic and social resources, and a concern for positive experiences of health care and outcomes. Health managers must ensure quality services, a functioning regulatory mechanism, and monitoring of provider behaviour.


Ozone detoxification of steam-pretreated Norway spruce.
Pretreatment of lignocellulose for biochemical conversion commonly results in formation of by-products that inhibit microorganisms and cellulolytic enzymes. To make bioconversion processes more efficient, inhibition problems can be alleviated through conditioning. Ozone is currently commercially employed in pulp and paper production for bleaching, as it offers the desirable capability to disrupt unsaturated bonds in lignin through an ionic reaction known as ozonolysis. Ozonolysis is more selective towards lignin than cellulose, for instance, when compared to other oxidative treatment methods, such as Fenton's reagent. Ozone may thus have desirable properties for conditioning of pretreated lignocellulose without concomitant degradation of cellulose or sugars. Ozone treatment of SO2-impregnated steam-pretreated Norway spruce was explored as a potential approach to decrease inhibition of yeast and cellulolytic enzymes. This novel approach was furthermore compared to some of the most effective methods for conditioning of pretreated lignocellulose, i.e., treatment with alkali and sodium dithionite. Low dosages of ozone decreased the total contents of phenolics to about half of the initial value and improved the fermentability. Increasing ozone dosages led to almost proportional increase in the contents of total acids, including formic acid, which ultimately led to poor fermentability at higher ozone dosages. The decrease of the contents of furfural and 5-hydroxymethylfurfural was inversely proportional (R (2) > 0.99) to the duration of the ozone treatment, but exhibited no connection with the fermentability. Ozone detoxification was compared with other detoxification methods and was superior to treatment with Fenton's reagent, which exhibited no positive effect on fermentability. However, ozone detoxification was less efficient than treatment with alkali or sodium dithionite. High ozone dosages decreased the inhibition of cellulolytic enzymes as the glucose yield was improved with 13 % compared to that of an untreated control. Low dosages of ozone were beneficial for the fermentation of steam-pretreated Norway spruce, while high dosages decreased the inhibition of cellulolytic enzymes by soluble components in the pretreatment liquid. While clearly of interest for conditioning of lignocellulosic hydrolysates, future challenges include finding conditions that provide beneficial effects both with regard to enzymatic saccharification and microbial fermentation.


CD8highCD57+ T lymphocytes in normal, healthy individuals are oligoclonal and respond to human cytomegalovirus.
CD8+CD57+ lymphocytes from normal peripheral blood are divided into T cells expressing high levels of CD8 and NK cells expressing low levels of surface CD8. Increased numbers of CD8highCD57+ T cells correlate with previous exposure to human cytomegalovirus (HCMV) infection, but no virus-specific function or function for CD8highCD57+ cells has been recorded. We have studied the TCR repertoire and responses of the CD8highCD57+ population induced by virus-infected fibroblasts in healthy individuals. Using three-color flow cytometry of PBL, we detected restricted TCRBV usage in the CD8highCD57+ subset of 11/15 subjects, compared with 1/15 in the CD8+, CD57- subset. The results of anchored PCR and sequencing also showed oligoclonality of TCR; 40-70% of CD8highCD57+ lymphocytes (10-20% of total CD8+ T cells) were derived from single clones. Such expansions were stable with time and detected in one subject over a 2-yr period. Functionally, CD8highCD57+ lymphocytes proliferated strongly to HCMV-, but not HSV-, VZV-, or influenza-infected fibroblasts in an MHC-unrestricted manner in vitro, including preferential augmentation of particular in vivo oligoclonally expanded subpopulations. HCMV-specific MHC-restricted CTL were detected, but limiting dilution analysis showed that these were a minority (< 10%) and not the oligoclonal subsets. In contrast, depletion of oligoclonally expanded CD8highCD57+ subpopulations, resulted in the increase of HCMV-specific CTL, suggesting functional heterogeneity in these cells.


Effects of non-pharmaceutical interventions on COVID-19 cases, deaths, and demand for hospital services in the UK: a modelling study.
Non-pharmaceutical interventions have been implemented to reduce transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the UK. Projecting the size of an unmitigated epidemic and the potential effect of different control measures has been crucial to support evidence-based policy making during the early stages of the epidemic. This study assesses the potential impact of different control measures for mitigating the burden of COVID-19 in the UK. We used a stochastic age-structured transmission model to explore a range of intervention scenarios, tracking 66·4 million people aggregated to 186 county-level administrative units in England, Wales, Scotland, and Northern Ireland. The four base interventions modelled were school closures, physical distancing, shielding of people aged 70 years or older, and self-isolation of symptomatic cases. We also modelled the combination of these interventions, as well as a programme of intensive interventions with phased lockdown-type restrictions that substantially limited contacts outside of the home for repeated periods. We simulated different triggers for the introduction of interventions, and estimated the impact of varying adherence to interventions across counties. For each scenario, we projected estimated new cases over time, patients requiring inpatient and critical care (ie, admission to the intensive care units [ICU]) treatment, and deaths, and compared the effect of each intervention on the basic reproduction number, R0. We projected a median unmitigated burden of 23 million (95% prediction interval 13-30) clinical cases and 350 000 deaths (170 000-480 000) due to COVID-19 in the UK by December, 2021. We found that the four base interventions were each likely to decrease R0, but not sufficiently to prevent ICU demand from exceeding health service capacity. The combined intervention was more effective at reducing R0, but only lockdown periods were sufficient to bring R0 near or below 1; the most stringent lockdown scenario resulted in a projected 120 000 cases (46 000-700 000) and 50 000 deaths (9300-160 000). Intensive interventions with lockdown periods would need to be in place for a large proportion of the coming year to prevent health-care demand exceeding availability. The characteristics of SARS-CoV-2 mean that extreme measures are probably required to bring the epidemic under control and to prevent very large numbers of deaths and an excess of demand on hospital beds, especially those in ICUs. Medical Research Council.


Auricular acupuncture treatment for insomnia: a systematic review.
To review trials on the efficacy and safety of auricular acupuncture (AA) treatment for insomnia and to identify the most commonly used auricular acupoints for treating insomnia in the studies via a frequency analysis. The international electronic databases searched included: (1) AMED; (2) the Cochrane library; (3) CINAHL; (4) EMBASE; and (5) MEDLINE. Chinese electronic databases searched included: (1) VIP Information; (2) CBMdisc; and (3) CNKI. Any randomized controlled trials using AA as an intervention without using any co-interventions for insomnia were included. Studies using AA versus no treatment, placebo, sham AA, or Western medicine were included. Two (2) independent reviewers were responsible for data extraction and assessment. The efficacy of AA was estimated by the relative risk (RR) using a meta-analysis. Eight hundred and seventy eight (878) papers were searched. Six (6) trials (402 treated with AA among 673 participants) that met the inclusion criteria were retrieved. A meta-analysis showed that AA was chosen with a higher priority among the treatment subjects than among the controls (p < 0.05). The recovery and improvement rates produced by AA was significantly higher than those of diazepam (p < 0.05). The rate of success was higher when AA was used for enhancement of sleeping hours up to 6 hours in treatment subjects (p < 0.05). The efficacy of using Semen vaccariae ear seeds was better than that of the controls (p < 0.01); while magnetic pearls did not show statistical significance (p = 0.28). Six (6) commonly used auricular acupoints were Shenmen (100%), Heart (83.33%), Occiput (66.67%), Subcortex (50%), Brain and Kidney (each 33.33%, respectively). AA appears to be effective for treating insomnia. Because the trials were low quality, further clinical trials with higher design quality, longer duration of treatment, and longer follow-up should be conducted.


Calmodulin kinase pathway mediates the K+-induced increase in Gap junctional communication between mouse spinal cord astrocytes.
Astrocytes are coupled to one another by gap junction channels that allow the diffusion of ions and small molecules throughout the interconnected syncytium. In astrocytes, gap junctions are believed to participate in spatial buffering removing the focal excess of potassium resultant from intense neuronal activity by current loops through the syncytium and are also implicated in the propagation of astrocytic calcium waves, a form of extraneuronal signaling. Gap junctions can be modulated by several factors, including elevation of extracellular potassium concentration. Because K(+) elevation is a component of spinal cord injury, we evaluated the extent to which cultured spinal cord astrocytes is affected by K(+) levels and obtained evidence suggesting that a Ca(2+)-calmodulin (CaM) protein kinase is involved in the K(+)-induced increased coupling. Exposure of astrocytes to high K(+) solutions induced a dose-dependent increase in dye coupling; such increased coupling was greatly attenuated by reducing extracellular Ca(2+) concentration, prevented by nifedipine, and potentiated by Bay-K-8644. These results indicate that K(+)-induced increased coupling is mediated by a signaling pathway that is dependent on the influx of Ca(2+) through L-type Ca(2+) channels. Evidence supporting the participation of the CaM kinase pathway on K(+)-induced increased coupling was obtained in experiments showing that calmidazolium and KN-93 totally prevented the increase in dye and electrical coupling induced by high K(+) solutions. Because no changes in connexin43 expression levels or distribution were observed in astrocytes exposed to high K(+) solutions, we propose that the increased junctional communication is related to an increased number of active channels within gap junction plaques.


2-(hydroxymethyl)pyridinium dihydrogenphosphate.
The title compound, C(6)H(8)NO(+).H(2)PO(4)(-), consists of 2-(hydroxymethyl)pyridinium and dihydrogenphosphate ions. The dihydrogenphosphate moieties are linked into chains by pairs of P-O-H.O-P hydrogen bonds. The 2-(hydroxymethyl)pyridinium cations are connected to the dihydrogenphosphate units by O-H.O and N-H.O hydrogen bonds. Weak pi-pi interactions help to determine the interchain packing.


Intracardiac thrombosis in Behçet's Disease successfully treated with immunosuppressive agents: A case of vascular pathergy phenomenon.
Behçet's Disease (BD) is a rare multi-systemic inflammatory disorder classified as a systemic vasculitis of unknown aetiology. Vascular involvement occurs in approximately 5-51.6% cases, affecting venous and arterial vessels. Cardiac involvement is rare in BD (6%). There have been published approximately 93 cases of BD associated with intracardiac thrombosis, with different treatments and courses. We present a case of a 35-year-old spanish male that, after a percutaneous pharmacomechanical thrombectomy with venous stent placement, developed high fever and extensive venous thrombosis despite anticoagulation including intracardiac thrombosis (ICT) in the right ventricle and pulmonary embolism that leaded to the diagnosis of BD. The patient was successfully treated with immunosuppressants, achieving the complete resolution of ICT. We hypotesize that the endovenous procedure could have acted as a trigger for the posterior acute attack of the disease, representing a 'vascular pathergy phenomenon'. Vascular BD has to be suspected in cases of thrombosis recurrence despite correct anticoagulation, and intense immunosuppressive treatment should be considered.


Associations between daily food intake and excess adiposity in Irish adults: towards the development of food-based dietary guidelines for reducing the prevalence of overweight and obesity.
The prevalence of obesity has nearly doubled in Ireland since 1990 and over half of the population has a large waist circumference (WC). No food-based, dietary guidelines exist in Ireland for a reduction in the prevalence of body fat or obesity. To examine the association between daily food intake and categories of body mass index and WC for the development of dietary guidelines to combat obesity. Cross-sectional study of a random representative sample of 1379 adults aged 18-64 years from Northern Ireland and the Republic of Ireland. Weight, height and WC were measured according to standard procedures. Diet was assessed using a 7-day food diary from which 28 food groups were generated and entered into logistic regression analysis. Higher mean daily consumption of most of the 28 food groups was associated with an increased likelihood of being classified as obese or at waist action level 2, compared to normal weight and normal WC. The strongest associations were found for savoury snacks, butter and full fat spreads. Contrary to popular opinion, not one individual food group but rather a combination of many foods was associated with excess adiposity. Body mass index and WC in adults are strongly influenced by the amount of food consumed. Public health policies for a reduction in body fat and obesity may be more effective if the emphasis is placed on a reduction of food and beverages consumed as opposed to the traditional dietary recommendations for macronutrients.


Antibacterial and hydroxyapatite-forming coating for biomedical implants based on polypeptide-functionalized titania nanospikes.
Titanium (Ti)-based implants often suffer from detrimental bacterial adhesion and inefficient healing, so it is crucial to design a dual-functional coating that prevents bacterial infection and enhances bioactivity for a successful implant. Herein, we successfully devised a cationic polypeptide (Pep)-functionalized biomimetic nanostructure coating with superior activity, which could not only kill pathogenic bacteria rapidly and inhibit biofilm formation for up to two weeks, but also promote in situ hydroxyapatite (HAp) formation. Specifically, a titania (TiO2) nanospike coating (TNC) was fabricated by alkaline hydrothermal treatment firstly, followed by immobilization of rationally synthesized Pep via robust coordinative interactions, named TNPC. This coating was able to effectively kill (>99.9%) both Gram-positive Staphylococcus aureus (S. aureus) and Gram-negative Escherichia coli (E. coli) bacteria, while being non-toxic to murine MC3T3-E1 osteoblastic cells. Furthermore, the in vivo infection studies denoted that the adherent bacteria numbers on the TNPC implants were significantly reduced by 6 orders of magnitude than those on the pure Ti implants (p < 0.001). Importantly, in the presence of cationic amino groups and residual Ti-OH groups, substantial HAp deposition on the TNPC surface in Kokubo's simulated body fluid (SBF) occurred after 14 days. Altogether, our results support the clinical potential of this biomimetic dual-functional coating as a new approach with desirable antibacterial properties and HAp-forming ability in orthopedic and dental applications.


Dorsolateral prefrontal cortex and anterior cingulate cortex white matter alterations in late-life depression.
The dorsolateral prefrontal cortex (DLPFC) and anterior cingulate cortex (ACC) are critical for mood regulation. Alterations in the white matter connections of these regions may impair their role in mood regulation and increase the risk of developing depression. This study used diffusion tensor imaging to examine for white matter microstructural abnormalities of these regions and of central white matter structures in late-life depression. One hundred six elderly depressed subjects and eighty-four elderly nondepressed subjects underwent clinical assessment and diffusion tensor imaging. The apparent diffusion coefficient (ADC) and fractional anisotropy (FA) were measured in regions of interest placed in the white matter of the DLPFC, ACC, corpus callosum, and internal capsule. Differences between groups were assessed, controlling for age, sex, and total cerebral volume. After controlling for covariates, depressed subjects had significantly lower FA values in white matter of the right ACC, bilateral superior frontal gyri, and left middle frontal gyrus. There were no significant differences in ADC values. Lower FA, representing lower tissue organization, is observed in depressed elders in the DLPFC and right ACC. These findings support the hypothesis that altered connectivity between brain regions contributes to the risk of depression.


Efficacy and safety of extending intravenous tocilizumab intervals from 4 to 6 weeks in rheumatoid arthritis patients with good response to 4-week intervals.
A period of 4 weeks (w) has been recommended for rheumatoid arthritis (RA) patients as the interval between intravenous (IV) tocilizumab (TCZ, 8 mg/kg). In a previous paper, we showed the possibility that the interval between successive IV TCZ can be extended from 4 to 6 weeks in more than 60% of patients with low diseases activity (LDA) at 4-week intervals. Herein, we aimed to investigate the efficacy and safety of extending the interval from 4 to 6 weeks. A retrospective observational study was conducted by enrolling patients in whom the intervals of TCZ infusions could be extended from 4 to 6 weeks with an LDA for more than 2 years. We compared the efficacy and side effects of TCZ infusions at intervals of 4 and 6 weeks in a cohort of patients. We also examined serum lipid, platelet, IL-6, and trough TCZ levels. A total of 125 patients with an LDA at 4 weeks intervals were enrolled in this study, of which 78 patients maintained LDA at 6-week intervals of TCZ infusion. After extending the infusion intervals, the efficacy of the treatment was maintained, and the side effects decreased significantly. In addition, the levels of total cholesterol and triglyceride were returned to normal, and the serum trough levels of TCZ became undetectable at 6-week intervals. We proved that intervals between TCZ infusions can be extended from 4 to 6 weeks in more than 60% of RA patients along with a decrease in the side effects, thus suggesting the need to change the infusion intervals in suitable patients.


Partial least squares-discriminant analysis (PLS-DA) for classification of high-dimensional (HD) data: a review of contemporary practice strategies and knowledge gaps.
Partial least squares-discriminant analysis (PLS-DA) is a versatile algorithm that can be used for predictive and descriptive modelling as well as for discriminative variable selection. However, versatility is both a blessing and a curse and the user needs to optimize a wealth of parameters before reaching reliable and valid outcomes. Over the past two decades, PLS-DA has demonstrated great success in modelling high-dimensional datasets for diverse purposes, e.g. product authentication in food analysis, diseases classification in medical diagnosis, and evidence analysis in forensic science. Despite that, in practice, many users have yet to grasp the essence of constructing a valid and reliable PLS-DA model. As the technology progresses, across every discipline, datasets are evolving into a more complex form, i.e. multi-class, imbalanced and colossal. Indeed, the community is welcoming a new era called big data. In this context, the aim of the article is two-fold: (a) to review, outline and describe the contemporary PLS-DA modelling practice strategies, and (b) to critically discuss the respective knowledge gaps that have emerged in response to the present big data era. This work could complement other available reviews or tutorials on PLS-DA, to provide a timely and user-friendly guide to researchers, especially those working in applied research.


Compressible boundary-layer density cross sections by UV Rayleigh scattering.
Rayleigh scattering has been used to image density cross sections of a compressible boundary layer for the first time to our knowledge. These cross sections were made from a Mach 2.5 boundary layer by illuminating it with a UV laser sheet from an argon-fluoride laser (193 nm). Since Rayleigh scattering is linearly proportional to density, quantitative values are measured, which leads to new instantaneous and statistical information about the structure of this turbulent field.


Timing of Circumcision and Breastfeeding Initiation Among Newborn Boys.
Newborn circumcision, particularly in the first 24 hours of life, has been thought to adversely affect breastfeeding initiation. However, no studies specifically support an association between early circumcision and difficulty with breastfeeding initiation or maintenance. This study was designed to determine whether timing of newborn circumcision affects rates of exclusive breastfeeding during the first 2 weeks of life. A retrospective study of 797 newborn boys and their mothers was conducted at a large military hospital. Exclusion criteria included gestational age <38 weeks, multiple delivery, NICU admission, and absence of maternal intention to breastfeed. Multivariable logistic regression models examined the relationship between time of elective circumcision and exclusive breastfeeding at 3 time points: hospital discharge and the newborn and 2-week outpatient visits. Mean infant age at circumcision was 29.7 hours. Thirty-one percent were circumcised at <24 hours of age. Rates of exclusive breastfeeding were 66.8%, 64.1%, and 63.7% at hospital discharge and the newborn and 2-week outpatient visits, respectively. In the multivariable model, time of circumcision was not significantly associated with exclusive breastfeeding at hospital discharge (odds ratio [OR], 1.01; 95% confidence interval [CI], 0.99-1.00; P = .54), the newborn outpatient visit (OR, 1.00; 95% CI, 0.99-1.02; P = .84), or the 2-week outpatient visit (OR, 0.99; 95% CI 0.98-1.01; P = .44). There was no significant association between timing of elective newborn circumcision and exclusive breastfeeding in the first 2 weeks of life. In this setting, early circumcision of otherwise healthy, term boys had no deleterious effects on breastfeeding initiation or maintenance.


LRRK2 but not ATG16L1 is associated with Paneth cell defect in Japanese Crohn's disease patients.
BACKGROUND. Morphological patterns of Paneth cells are a prognostic biomarker in Western Crohn's disease (CD) patients, and are associated with autophagy-associated ATG16L1 and NOD2 variants. We hypothesized that genetic determinants of Paneth cell phenotype in other ethnic CD cohorts are distinct but also involved in autophagy. METHODS. We performed a hypothesis-driven analysis of 56 single nucleotide polymorphisms (SNPs) associated with CD susceptibility or known to affect Paneth cell function in 110 Japanese CD patients who underwent ileal resection. We subsequently performed a genome-wide association analysis. Paneth cell phenotype was determined by defensin-5 immunofluorescence. Selected genotype-Paneth cell defect correlations were compared to a Western CD cohort (n = 164). RESULTS. The average percentage of abnormal Paneth cells in Japanese CD was similar to Western CD (P = 0.87), and abnormal Paneth cell phenotype was also associated with early recurrence (P = 0.013). In contrast to Western CD, ATG16L1 T300A was not associated with Paneth cell defect in Japanese CD (P = 0.20). Among the 56 selected SNPs, only LRRK2 M2397T showed significant association with Paneth cell defect (P = 3.62 × 10-4), whereas in the Western CD cohort it was not (P = 0.76). Pathway analysis of LRRK2 and other candidate genes with P less than 5 × 10-4 showed connections with known CD susceptibility genes and links to autophagy and TNF-α networks. CONCLUSIONS. We found dichotomous effects of ATG16L1 and LRRK2 on Paneth cell defect between Japanese and Western CD. Genes affecting Paneth cell phenotype in Japanese CD were also associated with autophagy. Paneth cell phenotype also predicted prognosis in Japanese CD. FUNDING. Helmsley Charitable Trust, Doris Duke Foundation (grant 2014103), Japan Society for the Promotion of Science (KAKENHI grants JP15H04805 and JP15K15284), Crohn's and Colitis Foundation grant 274415, NIH (grants 1R56DK095820, K01DK109081, and UL1 TR000448).


Profiling of substrate specificities of 3C-like proteases from group 1, 2a, 2b, and 3 coronaviruses.
Coronaviruses (CoVs) can be classified into alphacoronavirus (group 1), betacoronavirus (group 2), and gammacoronavirus (group 3) based on diversity of the protein sequences. Their 3C-like protease (3CL(pro)), which catalyzes the proteolytic processing of the polyproteins for viral replication, is a potential target for anti-coronaviral infection. Here, we profiled the substrate specificities of 3CL(pro) from human CoV NL63 (group 1), human CoV OC43 (group 2a), severe acute respiratory syndrome coronavirus (SARS-CoV) (group 2b) and infectious bronchitis virus (IBV) (group 3), by measuring their activity against a substrate library of 19 × 8 of variants with single substitutions at P5 to P3' positions. The results were correlated with structural properties like side chain volume, hydrophobicity, and secondary structure propensities of substituting residues. All 3CL(pro) prefer Gln at P1 position, Leu at P2 position, basic residues at P3 position, small hydrophobic residues at P4 position, and small residues at P1' and P2' positions. Despite 3CL(pro) from different groups of CoVs share many similarities in substrate specificities, differences in substrate specificities were observed at P4 positions, with IBV 3CL(pro) prefers P4-Pro and SARS-CoV 3CL(pro) prefers P4-Val. By combining the most favorable residues at P3 to P5 positions, we identified super-active substrate sequences 'VARLQ↓SGF' that can be cleaved efficiently by all 3CL(pro) with relative activity of 1.7 to 3.2, and 'VPRLQ↓SGF' that can be cleaved specifically by IBV 3CL(pro) with relative activity of 4.3. The comprehensive substrate specificities of 3CL(pro) from each of the group 1, 2a, 2b, and 3 CoVs have been profiled in this study, which may provide insights into a rational design of broad-spectrum peptidomimetic inhibitors targeting the proteases.


Impacts and mitigation of excess diesel-related NOx emissions in 11 major vehicle markets.
Vehicle emissions contribute to fine particulate matter (PM2.5) and tropospheric ozone air pollution, affecting human health, crop yields and climate worldwide. On-road diesel vehicles produce approximately 20 per cent of global anthropogenic emissions of nitrogen oxides (NOx), which are key PM2.5 and ozone precursors. Regulated NOx emission limits in leading markets have been progressively tightened, but current diesel vehicles emit far more NOx under real-world operating conditions than during laboratory certification testing. Here we show that across 11 markets, representing approximately 80 per cent of global diesel vehicle sales, nearly one-third of on-road heavy-duty diesel vehicle emissions and over half of on-road light-duty diesel vehicle emissions are in excess of certification limits. These excess emissions (totalling 4.6 million tons) are associated with about 38,000 PM2.5- and ozone-related premature deaths globally in 2015, including about 10 per cent of all ozone-related premature deaths in the 28 European Union member states. Heavy-duty vehicles are the dominant contributor to excess diesel NOx emissions and associated health impacts in almost all regions. Adopting and enforcing next-generation standards (more stringent than Euro 6/VI) could nearly eliminate real-world diesel-related NOx emissions in these markets, avoiding approximately 174,000 global PM2.5- and ozone-related premature deaths in 2040. Most of these benefits can be achieved by implementing Euro VI standards where they have not yet been adopted for heavy-duty vehicles.


[Sexual and reproductive rights: challenges for health policies].
This article discusses sexual and reproductive rights in the sense of a stance that assumes a perspective of transformations in social relations, the struggle against prejudices, the guarantee of well-being, and finally the relationship between sexuality, reproduction, and citizenship. The article then proceeds to reflect on health policy challenges in these fields, emphasizing such issues as: guaranteed resources, quality and quantity of health services in response to demands by the population, and cultural changes that produce a new view of the relationship between health professionals and health system clients, based on the principles of citizenship: recognition others as entitled to freedom and equality.


Neuronal morphology generates high-frequency firing resonance.
The attenuation of neuronal voltage responses to high-frequency current inputs by the membrane capacitance is believed to limit single-cell bandwidth. However, neuronal populations subject to stochastic fluctuations can follow inputs beyond this limit. We investigated this apparent paradox theoretically and experimentally using Purkinje cells in the cerebellum, a motor structure that benefits from rapid information transfer. We analyzed the modulation of firing in response to the somatic injection of sinusoidal currents. Computational modeling suggested that, instead of decreasing with frequency, modulation amplitude can increase up to high frequencies because of cellular morphology. Electrophysiological measurements in adult rat slices confirmed this prediction and displayed a marked resonance at 200 Hz. We elucidated the underlying mechanism, showing that the two-compartment morphology of the Purkinje cell, interacting with a simple spiking mechanism and dendritic fluctuations, is sufficient to create high-frequency signal amplification. This mechanism, which we term morphology-induced resonance, is selective for somatic inputs, which in the Purkinje cell are exclusively inhibitory. The resonance sensitizes Purkinje cells in the frequency range of population oscillations observed in vivo.


Employment of intra-individual variability to improve computerized ECG interpretation.
One of the reasons for the limited practical utility of computer programs for interpretation of electrocardiograms (ECGs) is their susceptibility to intra-individual variability. Two of the most prominent sources of intra-individual variability in ECGs, electrode placement variations and respiration, were studied for their effects on computerized ECG interpretation. Previous research has shown that the effects of intra-individual variability on computerized ECG interpretation depend largely on the individual ECG. To enable the assessment of chest electrode position variations for individual standard 12- lead ECGs, ECGs resulting from simulations of such position variations were interpreted. Variability due to respiration was assessed by interpreting all individual ECG beats instead of an averaged beat. In this paper two methods are presented that employ information about the intra-individual variability in individual ECGs. The first method provides an estimate of the reliability of the interpretation, the second attempts to improve the interpretation itself. In the first method we quantified the variation in interpretation caused by the two sources of intra-individual variability with the use of a stability index, a high index value indicating a low variation in interpretation. This index was subsequently studied using two sets of ECGs. For the first set a â clinical' reference interpretation was obtained from discharge letters. For the second set three cardiologists provided a â cardiologists' reference. The performance of subgroups of ECGs having stability indices higher than a particular value was computed. It appeared that for the â cardiologists' reference, the interpretations of ECGs with a high stability index were more often correct. No effect was found for the â clinical' reference. In the second method we attempted to improve the original interpretation by combining the alternative interpretations into a new interpretation. This was done by taking the median or the average of the quantified alternatives. These combined interpretations proved to perform better than the original interpretation when a cardiologist's interpretation was taken as a reference. This paper shows that intra-individual ECG variability can be used to improve original interpretations. This can be done without having to record multiple ECGs, provided that a model is available to simulate intra-individual variability. The presented methods do not depend on the classification algorithm that is used. They can be used both during classifier design to correct imperfections, and in routine use of the classifier to produce more representative classifications.


Threshold of Pulmonary Hypertension Associated With Increased Mortality.
There is increasing evidence that current thresholds for diagnosing pulmonary hypertension (PHT) underestimate the prognostic impact of PHT. The aim of this study was to determine the prognostic impact of increasing pulmonary pressures within the National Echocardiography Database of Australia cohort (n = 313,492). The distribution of estimated right ventricular systolic pressure (eRVSP) was examined in 157,842 men and women. All had data linkage to long-term survival during median follow-up of 4.2 years (interquartile range: 2.2 to 7.5 years). The cohort comprised 74,405 men and 83,437 women 65.6 ± 17.7 years of age. Overall, 17,955 (11.4%), 7,016 (4.4%), and 4,515 (2.9%) subjects had eRVSP levels indicative of mild (40 to 49 mm Hg), moderate (50 to 59 mm Hg), or severe (≥60 mm Hg) PHT, respectively, assuming a right atrial pressure of 5 mm Hg. These subjects were more likely to die during long-term follow up (for severe PHT, adjusted hazard ratio: 9.73; 95% confidence interval: 8.60 to 11.0; p < 0.001). After adjustment for age, sex, and evidence of left heart disease, those subjects with eRVSP levels within the third (28.05 to 32.0 mm Hg; hazard ratio: 1.410; 95% confidence interval: 1.310 to 1.517) and fourth (32.05 to 38.83 mm Hg; hazard ratio: 1.979; 95% confidence interval: 1.853 to 2.114) quintiles had significantly higher mortality (p < 0.001) than those in the lowest quintile. Accordingly, a clear and consistent threshold of increased mortality (including 1- and 5-year actuarial mortality) around an eRVSP of 30.0 mm Hg was evident. In this large and unique cohort, the prognostic impact of clinically accepted levels of PHT was confirmed. Moreover, a distinctly lower threshold for increased risk for mortality (eRVSP >30.0 mm Hg) indicative of PHT was identified. (A Longitudinal Cohort Study of Echocardiograms From Public and Private Echocardiography Laboratories From Around Australia, Linked With the National Deaths Index; ACTRN12617001387314).


[Continuing hospice care of cancer--a three-year experience].
The hospice at Mackay Memorial Hospital was established in February 1990. A group of team workers including physicians, nurses, social workers and the clergy were involved in this holistic care program for terminal cancer patients. Four hundred and seventy-nine patients were eligible for the program up to February 1993. Regarding duration of stay, 62.5% of patients resided for 14 days. Those surviving under 90 days constituted 75.5% of patients. Fifty-one point eight percent of patients died in the hospice and 18.2% died at home soon after being discharged from the hospice. Pain is the most common symptom among the patients. Treatment strategies vary according to the three-step-ladder protocol designed by WHO. Total pain relief was achieved in 80% of patients. Opportune private talking and family conferences formed the basis of the "peer model". Through this model, treatment decisions including physical, psychosocial and spiritual issues were made. Before the peer model, only 36 (10.3%) patients agreed with the idea of hospice care, while 257 (73.6%) patients agreed after the model was established. Awareness of dying was evident in 412 (86%) patients. Two hundred and eighty (68%) patients became aware of the prospect of death through guessing, while the other 132 (32%) patients were informed by medical staff. Problems encountered by the team workers included 1) needs in education and training, 2) psychological pressure, 3) management of loss and grief, 4) needs in supportive system and 5) troubles caused by families' lying to patients. The team workers were satisfied with the quality of care in 38.4% of patients and fairly satisfied with 30.7% of patients.(ABSTRACT TRUNCATED AT 250 WORDS)


Applying Unique Molecular Identifiers in Next Generation Sequencing Reveals a Constrained Viral Quasispecies Evolution under Cross-Reactive Antibody Pressure Targeting Long Alpha Helix of Hemagglutinin.
To overcome yearly efforts and costs for the production of seasonal influenza vaccines, new approaches for the induction of broadly protective and long-lasting immune responses have been developed in the past decade. To warrant safety and efficacy of the emerging crossreactive vaccine candidates, it is critical to understand the evolution of influenza viruses in response to these new immune pressures. Here we applied unique molecular identifiers in next generation sequencing to analyze the evolution of influenza quasispecies under in vivo antibody pressure targeting the hemagglutinin (HA) long alpha helix (LAH). Our vaccine targeting LAH of hemagglutinin elicited significant seroconversion and protection against homologous and heterologous influenza virus strains in mice. The vaccine not only significantly reduced lung viral titers, but also induced a well-known bottleneck effect by decreasing virus diversity. In contrast to the classical bottleneck effect, here we showed a significant increase in the frequency of viruses with amino acid sequences identical to that of vaccine targeting LAH domain. No escape mutant emerged after vaccination. These results not only support the potential of a universal influenza vaccine targeting the conserved LAH domains, but also clearly demonstrate that the well-established bottleneck effect on viral quasispecies evolution does not necessarily generate escape mutants.


Resolution and registration in dual-plane co-RASOR MR.
Magnetic resonance imaging (MRI) has superior soft tissue contrast and lower interobserver variability compared to computed tomography and advances in equipment and pseudo-CT estimation have allowed for MR-only radiation therapy planning. Dedicated MR sequences have been used to localize paramagnetic structures with positive contrast, and most implanted seeds are gold fiducial markers (GFMs). We used a fast, dual-plane co-RASOR sequence to localize implanted GFMs with positive contrast in phantom and tissue to assess their resolution and registration accuracy of registration to CT. Off-resonant reconstructions of co-RASOR images were able to resolve GFMs down to 5 mm apart at 12 cm FOV. No systematic biases were observed by comparing registration of co-RASOR and bSSFP to CT images in an MR-compatible Lego phantom with a set of highly visible known points. The standard deviations of the MR to CT distance errors were <0.5 mm in all directions. We separated the component due to registration by comparing the two MR sequences, which had a maximum standard deviation of 0.36 mm in the SI-direction. Registration using the positive contrast points in a porcine sample phantom showed increased errors, but co-RASOR still performs acceptably with a target registration error of <0.75 mm. The dual-plane co-RASOR sequence could then be used for both registration and image tracking when performing MR-only radiation therapy planning.


Triple-marker cardiac MRI detects sequential tissue changes of healing myocardium after a hydrogel-based therapy.
Regenerative therapies based on injectable biomaterials, hold an unparalleled potential for treating myocardial ischemia. Yet, noninvasive evaluation of their efficacy has been lagging behind. Here, we report the development and longitudinal application of multiparametric cardiac magnetic resonance imaging (MRI) to evaluate a hydrogel-based cardiac regenerative therapy. A pH-switchable hydrogel was loaded with slow releasing insulin growth factor 1 and vascular endothelial growth factor, followed by intramyocardial injection in a mouse model of ischemia reperfusion injury. Longitudinal cardiac MRI assessed three hallmarks of cardiac regeneration: angiogenesis, resolution of fibrosis and (re)muscularization after infarction. The multiparametric approach contained dynamic contrast enhanced MRI that measured improved vessel features by assessing fractional blood volume and permeability*surface area product, T1-mapping that displayed reduced fibrosis, and tagging MRI that showed improved regional myocardial strain in hydrogel treated infarcts. Finally, standard volumetric MRI demonstrated improved left ventricular functioning in hydrogel treated mice followed over time. Histology confirmed MR-based vessel features and fibrotic measurements. Our novel triple-marker strategy enabled detection of ameliorated regeneration in hydrogel treated hearts highlighting the translational potential of these longitudinal MRI approaches.


The C2 domains of the class I Rab11 family of interacting proteins target recycling vesicles to the plasma membrane.
The Rab11 family of interacting proteins (Rab11-FIP) is a recently identified protein family composed of, to date, six members that interact with Rab11. They all share a highly homologous Rab11-binding domain (RBD) at their C-termini. However, apart from the RBD, they vary in their domain organization. Rab11-FIP3 and Rab11-FIP4 possess an ezrin-radixin-moesin (ERM) domain in their C-terminal half and EF hands in their N-terminal region. They have been termed class II Rab11-FIPs. The class I Rab11-FIPs, Rab coupling protein (RCP), Rip11 and Rab11-FIP2, each have a C2 phospholipid-binding domain near their N-termini. Although they are still membrane associated, truncation mutants of the class I Rab11-FIPs that lack their C2 domains display an altered subcellular distribution in vivo, indicating that this domain plays an important role in specifying their correct intracellular localization. To determine the phospholipids to which they bind, a protein phospholipid overlay assay was performed. Our results indicate that the class-I Rab11-FIPs bind preferentially to phosphatidylinositol-(3,4,5)-trisphosphate [PtdIns(3,4,5)P3] and the second messenger phosphatidic acid. Stimulation of PtdIns(3,4,5)P3 or phosphatidic acid synthesis results in the translocation of the Rab11-FIPs from a perinuclear location to the periphery of the cell. By contrast, the transferrin receptor does not translocate to the plasma membrane under these conditions. This translocation is dependent on the presence of the C2 domain, because class I Rab11-FIP green-fluorescent-protein fusions that lack the C2 domain cannot translocate to the plasma membrane. We propose that the C2 domains of the class I Rab11-FIPs function to target these proteins to 'docking sites' in the plasma membrane that are enriched in PtdIns(3,4,5)P3 and phosphatidic acid.


Spatial-spectral volume holographic systems: resolution dependence on effective thickness.
The resolution dependence of spatial-spectral volume holographic imaging systems on angular and spectral bandwidth of nonuniform gratings is investigated. Modeling techniques include a combination of the approximate coupled-wave analysis and the transfer-matrix method for holograms recorded in absorptive media. The effective thickness of the holograms is used as an estimator of the resolution of the imaging systems. The methodology, which assists in the design and optimization of volume holographic simulation results based on our approach, are confirmed with experiments and show proof of consistency and usefulness of the proposed models.


Brief Internet-based intervention reduces posttraumatic stress and prolonged grief in parents after the loss of a child during pregnancy: a randomized controlled trial.
The loss of a child during pregnancy causes significant psychological distress for many women and their partners, and may lead to long-lasting psychiatric disorders. Internet-based interventions using exposure techniques and cognitive restructuring have proved effective for posttraumatic stress disorder (PTSD) and prolonged grief. This study compared the effects of an Internet-based intervention for parents after prenatal loss with a waiting list condition (WLC). The Impact of Event Scale - Revised assessed symptoms of PTSD; the Inventory of Complicated Grief and the Brief Symptom Inventory assessed depression, anxiety, and general mental health. The 228 participants (92% female) were randomly allocated to a treatment group (TG; n = 115) or a WLC group (n = 113). The TG received a 5-week cognitive behavioral intervention including (1) self-confrontation, (2) cognitive restructuring, and (3) social sharing. The TG showed significantly reduced symptoms of posttraumatic stress, prolonged grief, depression, and anxiety relative to the WLC control group. Intention-to-treat analysis revealed treatment effects of between d = 0.84 and d = 1.02 for posttraumatic stress and prolonged grief from pre- to posttreatment time points. Further significant improvement in all symptoms of PTSD and prolonged grief was found from the posttreatment evaluation to the 12-month follow-up. The attrition rate of 14% was relatively low. The Internet-based intervention proved to be a feasible and cost-effective treatment, reducing symptoms of posttraumatic stress, grief, depression, anxiety, and general mental health after pregnancy loss. Low-threshold e-health interventions should be further evaluated and implemented routinely to improve psychological support after pregnancy loss.


Intereye Comparison of Lamina Cribrosa Curvature in Normal Tension Glaucoma Patients With Unilateral Damage.
To investigate intereye differences in lamina cribrosa (LC) morphology in normal tension glaucoma (NTG) patients with unilateral damage. A total of 152 eyes of 76 treatment-naive NTG patients with unilateral damage from the ongoing Investigating Glaucoma Progression Study were included. Optic nerve heads were scanned using enhanced-depth spectral-domain optical coherence tomography. The magnitude of the LC curve and LC position were assessed by measuring the LC curve index (LCCI) and LC depth (LCD), respectively, at seven locations spaced equidistantly across the vertical optic disc diameter. LCCI and LCD were compared between glaucomatous and fellow healthy eyes. Eyes with NTG had larger average LCCI and LCD than contralateral healthy eyes (for both P < 0.002). The LCCI was greater in the glaucomatous eyes at all seven locations (P < 0.001). Univariate conditional logistic regression analysis showed that higher baseline intraocular pressure (P = 0.010), deeper LCD (P = 0.007), and larger LCCI (P < 0.001) were significantly associated with the presence of glaucoma. In multivariate analysis, only larger LCCI was significantly associated with the presence of glaucoma (P < 0.001). Glaucomatous eyes have more steeply curved LC than fellow healthy eyes. This finding suggests that LC undergoes significant remodeling in NTG eyes.


An inducible knockout of Dicer in adult mice does not affect endurance exercise-induced muscle adaptation.
The contractile and metabolic properties of adult skeletal muscle change in response to endurance exercise. The mechanisms of transcriptional regulation in exercise-induced skeletal muscle adaptation, including fiber-type switching and mitochondrial biogenesis, have been investigated intensively, whereas the role of microRNA (miRNA)-mediated posttranscriptional gene regulation is less well understood. We used tamoxifen-inducible Dicer1 knockout (iDicer KO) mice to reduce the global expression of miRNAs in adult skeletal muscle and subjected these mice to 2 wk of voluntary wheel running. Dicer mRNA expression was completely depleted in fast-twitch plantaris muscle after tamoxifen injection. However, several muscle-enriched miRNAs, including miR-1 and miR-133a, were reduced by only 30-50% in both the slow and fast muscles. The endurance exercise-induced changes that occurred for many parameters (i.e., fast-to-slow fiber-type switch and increases in succinate dehydrogenase, respiratory chain complex II, and citrate synthase activity) in wild type (WT) also occurred in the iDicer KO mice. Protein expression of myosin heavy chain IIa, peroxisome proliferator-activated receptor-γ coactivator-1α, and cytochrome c complex IV was also increased in the iDicer KO mice by the voluntary running. Furthermore, there was no significant difference in oxygen consumption rate in the isolated mitochondria between the WT and iDicer KO mice. These data indicate that muscle-enriched miRNAs were detectable even after 4 wk of tamoxifen treatment and there was no apparent specific endurance-exercise-induced muscle phenotype in the iDicer KO mice.


Effects of intra- and interpatch host density on egg parasitism by three species of Trichogramma.
Host-foraging responses to different intra- and interpatch densities were used to assess three Trichogramma spp. (Hymenoptera: Trichogrammatidae) Trichogramma deion Pinto and Oatman, T. ostriniae Pang and Chen, and T. pretiosum Riley - as potential biological control agents for the Indian meal moth, Plodia interpunctella Hübner (Lepidoptera: Pyralidae). Single naïve females were allowed 6 h to forage in Plexiglas arenas with four different spatial arrangements of host eggs, nine single-egg patches), nine four-egg patches, 36 single-egg patches, and 36 four-egg patches. No significant differences were found among species in the number of patches parasitized. As expected, all three species parasitized the most eggs in the 36 four-egg patch treatment and the least in the nine single-egg patch treatment. T. deion parasitized significantly more eggs than T. pretiosum on the nine four-egg patches. T. ostriniae parasitized significantly more patches when intrapatch density was greater, regardless of interpatch density. In contrast, T. deion only parasitized more patches at the greater intrapatch density when the interpatch density was low. Patch density had no effect on T. pretiosum. The spatial pattern of parasitism was more aggregated for T. deion and T. ostriniae in the 36 four-egg patches treatment compared to the 36 single-egg patches treatment. Therefore, intrapatch density was more important than interpatch density for T. ostriniae, and potentially for T. deion, but not for T. pretiosum. T. deion may be the best candidate for augmentative biological control because it parasitized either slightly or significantly more eggs than the other two species in all four treatments. Furthermore, the pattern of parasitism by T. deion in the 36 four-egg patches treatment was the most aggregated among the three species, suggesting a more thorough searching pattern. In contrast, T. pretiosum had the least aggregated pattern of parasitism and therefore may have used a more random foraging pattern.


Primary care patients' views and decisions about, experience of and reactions to direct-to-consumer genetic testing: a longitudinal study.
Little is known about the decisions and perspectives of participants undergoing direct-to-consumer genetic testing (DTCGT). The aims of this study were to examine the views, attitudes and decision-making factors of primary care patients regarding DTCGT. Their experience of and reactions to testing also emerged during the study. In this longitudinal, qualitative study, 20 primary care patients participated in DTCGT and individual interviews: (1) prior to testing after the informed consent session, (2) after receiving results, (3) 3 months post-test, and (4) 12 months post-test. Interviews included open-ended questions and all transcripts were analyzed using grounded theory, constant comparison methods. Five key themes emerged from data analysis as participants underwent DTCGT and reflected on their decision over time: (1) limited concerns about DTCGT, (2) motivations for testing, (3) expectations of testing, (4) understanding of results, and (5) impact of testing and results. While a few participants expressed concerns before testing, participants were motivated to test by curiosity, gaining actionable knowledge, and altruism. Most were uncertain of what to expect from DTCGT and needed assistance in understanding results. While many reported testing had no significant impact on them, being relieved or pleased after testing was the most common emotional effect. Notably, a few participants made positive health changes in response to testing. Given the paucity of information about primary care patients and DTCGT, this study adds more in-depth information to the emerging research on how such participants' view, make decisions about, experience and react to DTCGT over time. Because uncertainty remains about the accuracy of DTCGT, the response of primary care patients to this testing requires further investigation.


Lessons from the pilot of a mobile application to map assistive technology suppliers in Africa.
A pilot project to develop and implement a mobile smartphone application (App) that tracks and maps assistive technology (AT) availability in southern Africa was launched in Botswana in 2016. The App was developed and tested through an iterative process. The concept of the App (AT-Info-Map) was well received by most stakeholders within the pilot country, and broader networks. Several technical and logistical obstacles were encountered. These included high data costs; difficulty in accessing AT information from the public healthcare sector, the largest supplier of AT; and the high human resource demand of collecting and keeping up-to-date device-level information within a complex and fragmented supply sector that spans private, public and civil society entities. The challenges were dealt with by keeping the data burden low and eliminating product-level tracking. The App design was expanded to include disability services, contextually specific AT categories and make navigation more intuitive. Long-term sustainability strategies like generating funding through advertisements on the App or supplier usage fees must be explored. Outreach and sensitisation programmes about both the App and AT in general must be intensified. The project team must continually strengthen partnerships with private and public stakeholders to ensure ongoing project engagement. The lessons learnt might be of value to others who wish to embark on initiatives in AT and/or implement Apps in health or disability in southern Africa and in low-resourced settings around the world.


Flash scanning electron microscopy.
Scanning Electron Microscopy (SEM) is an invaluable tool for biologists and neuroscientists to study brain structure at the intracellular level. While able to image tissue samples with up to 5 nm isotropic resolution, image acquisition is prohibitively slow and limits the size of processed samples. In this work, we propose a novel approach to speeding up imaging when looking for specific structures. Unlike earlier methods, we explicitly balance the conflicting requirements of spending enough time scanning potential regions of interest to ensure that all targets are found while not wasting time on unpromising regions. This is achieved by using a Markov Random Field to model target locations and optimizing scanning locations by using a Branch-and-Bound strategy. We show that our approach significantly outperforms state-of-the-art methods to locate mitochondria in brain tissue.


Community Pharmacists' Views and Practices Regarding Natural Health Products Sold in Community Pharmacies.
Reports of regulatory and evidentiary gaps have raised concerns about the marketing and use of natural health products (NHPs). The majority of NHPs offered for sale are purchased at a community pharmacy and pharmacists are "front-line" health professionals involved in the marketing and provision of NHPs. To date, the involvement of pharmacists in pharmacy care involving NHPs and the degree to which concerns over the safety, efficacy, marketing and regulation of NHPs are addressed in pharmacy care in Canada have not been studied. Using Qualtrics, a web-based data collection and analysis software, and a study instrument made up of fifteen (15) open-ended, closed and rating scale questions, we surveyed the attitudes and practices of 403 community pharmacists in the Canadian province of Alberta regarding NHPs offered for sale in community pharmacies. The majority of pharmacists surveyed (276; 68%) recommend NHPs to clients sometimes to very often. Vitamin D, calcium, multivitamins, prenatal vitamins, probiotics and fish oil and omega-3 fatty acids were the most frequently recommended NHPs. The most common indications for which NHPs are recommended include bone and musculoskeletal disorders, maintenance of general health, gastrointestinal disorders and pregnancy. Review articles published in the Pharmacist's Letter and Canadian Pharmacists Journal were the primary basis for recommending NHPs. The majority of pharmacists surveyed (339; 84%) recommend the use of NHPs concurrently with conventional drugs, while a significant number and proportion (125; 31%) recommend alternative use. Pharmacists in the study overwhelmingly reported providing counselling on NHPs to clients based on information obtained mainly from the Natural Medicines Comprehensive Database. The study findings indicate a high prevalence of pharmacy care relating to NHPs among study participants. Although pharmacists' practices around NHPs are consistent with the existing licensing framework, we found some involvement in problematic practices that necessitate further research and potential policy scrutiny. The study also uncovered patterns of recommendations, including sources relied on in recommending NHPs and in providing counselling to patients, that raise concerns about the quality and credibility of NHP-related care provided to pharmacy patrons.


Alpha-CaMKII plays a critical role in determining the aggressive behavior of human osteosarcoma.
Osteosarcoma is among the most frequently occurring primary bone tumors, primarily affecting adolescents and young adults. Despite improvements in osteosarcoma treatment, more specific molecular targets are needed as potential therapeutic options. One target of interest is α-Ca(2+)/calmodulin-dependent protein kinase II (α-CaMKII), a ubiquitous mediator of Ca(2+)-linked signaling, which has been shown to regulate tumor cell proliferation and differentiation. Here, we investigate the role of α-CaMKII in the growth and tumorigenicity of human osteosarcoma. We show that α-CaMKII is highly expressed in primary osteosarcoma tissue derived from 114 patients, and is expressed in varying levels in different human osteosarcoma (OS) cell lines [MG-63, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)/HOS, and 143B). To examine whether α-CaMKII regulates osteosarcoma tumorigenic properties, we genetically inhibited α-CaMKII in two osteosarcoma cell lines using two different α-CaMKII shRNAs delivered by lentiviral vectors and overexpressed α-CaMKII by retrovirus. The genetic deletion of α-CaMKII by short hairpin RNA (shRNA) in MG-63 and 143B cells resulted in decreased proliferation (50% and 41%), migration (22% and 25%), and invasion (95% and 90%), respectively. The overexpression of α-CaMKII in HOS cells resulted in increased proliferation (240%), migration (640%), and invasion (10,000%). Furthermore, α-CaMKII deletion in MG-63 cells significantly reduced tumor burden in vivo (65%), whereas α-CaMKII overexpression resulted in tumor formation in a previously nontumor forming osteosarcoma cell line (HOS). Our results suggest that α-CaMKII plays a critical role in determining the aggressive phenotype of osteosarcoma, and its inhibition could be an attractive therapeutic target to combat this devastating adolescent disease.


Monitoring of a biogas process using electronic gas sensors and near-infrared spectroscopy (NIR).
The use of electronic gas sensors and near-infrared spectroscopy (NIR) to monitor the dynamics in a biogas process was evaluated using multivariate data analysis. The digester, a completely stirred 8 l tank reactor fed with a mixture of cellulose, albumin and minerals, was exposed to an overload of glucose after which monitoring of electronic gas sensor responses, NIR spectra as well as traditional chemical variables and analysis of microbial community structure were done. The responses from an array of electronic gas sensors consisting of MOS and MOSFET-sensors were correlated against volatile compounds in the headspace using partial least square (PLS) regressions. The root mean square error of prediction (RMSEP) was 0.15 g/l for acetate in the range of 0.14-1.72 g/l and the RMSEP for methane was 2.3% in the range of 27-73%. Selected wavelengths from the second derivative of the original NIR spectra (400-2500 nm) resulted in a PLS-model for predicting microbial biomass, measured as total phospholipid fatty acids, with a RMSEP of 9 nmol/ml in the range of 163-293 nmol/ml. The NIR model developed for acetate had a RMSEP of 0.20 g/l within the range of 0.14-1.72 g/l. The results clearly show that both NIR and an array of electronic gas sensors can provide simultaneous non-invasive in situ monitoring of important process variables in anaerobic digesters.


The arthroscopic Latarjet: a multisurgeon learning curve analysis.
The open Latarjet procedure is a standard surgical treatment option for anterior shoulder instability in patients with a high risk of failure following soft tissue stabilization. The arthroscopic technique has potential advantages of minimal invasiveness, reduced postoperative stiffness, and faster rehabilitation but is regarded as technically challenging with concern over surgical risk during the learning curve. The aim of this study was to undertake a multisurgeon, large-volume learning curve analysis of the arthroscopic Latarjet procedure using continuous learning curve analysis. Individual patient data from 12 surgeons across 5 countries were retrospectively reviewed. A total of 573 patients undergoing the arthroscopic Latarjet procedure were included. Outcome measures of learning were collected, including operative time, computed tomography (CT) bone-block positioning, complications, and patient-reported outcome measures (PROMs). A segmented linear regression modeling technique was used for learning curve analysis. High-volume surgeons converged to an operative time steady state after 30-50 cases. Surgeons completing fewer procedures demonstrated a constant reduction in operative time without reaching a plateau. Low-volume surgeons completing fewer than 14 operations did not demonstrate a reduction in operative time. Accuracy of bone-block positioning on postoperative CT demonstrated constant improvement without reaching a plateau after 53 cases. There was no change in PROMs or complications with increased operative volume. Specialist shoulder surgeons require 30-50 arthroscopic Latarjet procedures to attain steady-state operative efficiency, during which there is improvement in bone-block positioning. Only surgeons expecting to undertake the arthroscopic Latarjet in high volume should consider adopting this procedure.


Clinical biomechanic correlates for cervical function: Part III. Intermittent secondary movements.
Parts I and II of this study compared kinematic and myoelectric data for two groups of asymptomatic subjects classified as having symmetric or asymmetric motor response to a palpatory test for cervical sidebending. Kinematic data revealed that asymmetric subjects had limited mobility for primary and secondary motions. Myoelectric activity was slow to be initiated in the asymmetric subjects, and reduced in time and strength of contraction. Part III addresses additional specific kinematic data concerning three-dimensional orientations of the head; however, these data were accumulated throughout the paths of movement, not just at their end points as in our previous work. Although asymmetric subjects had demonstrated significantly reduced range of motion, paths of the more minor secondary axes did not differ significantly between groups. As with previous data (Part I and II), active and passive movements were undifferentiated; this degree of likeness in even minor aspects of the motor performance continues to indicate the remarkable similarities that can exist between primary movements directed and guided by trained physicians and those actively controlled by patients themselves. Part III completes a study of cervical motor behavior in which a passive gross motion test distinguished an asymmetric group with subclinical motor behavior that has measurable kinematic and myoelectric correlates.


Astaxanthin protects against MPTP/MPP+-induced mitochondrial dysfunction and ROS production in vivo and in vitro.
Astaxanthin (AST) is a powerful antioxidant that occurs naturally in a wide variety of living organisms. We have investigated the role of AST in preventing 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced apoptosis of the substantia nigra (SN) neurons in the mouse model of Parkinson's disease (PD) and 1-methyl-4-phenylpyridinium (MPP+)-induced cytotoxicity of SH-SY5Y human neuroblastoma cells. In in vitro study, AST inhibits MPP+-induced production of intracellular reactive oxygen species (ROS) and cytotoxicity in SH-SY5Y human neuroblastoma cells. Preincubation of AST (50 μM) significantly attenuates MPP+-induced oxidative damage. Furthermore, AST is able to enhance the expression of Bcl-2 protein but reduce the expression of α-synuclein and Bax, and suppress the cleavage of caspase-3. Our results suggest that the protective effects of AST on MPP+-induced apoptosis may be due to its anti-oxidative properties and anti-apoptotic activity via induction of expression of superoxide dismutase (SOD) and catalase and regulating the expression of Bcl-2 and Bax. Pretreatment with AST (30 mg/kg) markedly increases tyrosine hydroxylase (TH)-positive neurons and decreases the argyrophilic neurons compared with the MPTP model group. In summary, AST shows protection from MPP+/MPTP-induced apoptosis in the SH-SY5Y cells and PD model mouse SN neurons, and this effect may be attributable to upregulation of the expression of Bcl-2 protein, downregulation of the expression of Bax and α-synuclein, and inhibition of the activation of caspase-3. These data indicate that AST may provide a valuable therapeutic strategy for the treatment of progressive neurodegenerative disease such as Parkinson's disease.


Reported Sports Participation, Race, Sex, Ethnicity, and Obesity in US Adolescents From NHANES Physical Activity (PAQ_D).
Objective. To understand the relationships between participation in different types of leisure time sport activity and adolescent obesity, and how those relationships might differ based on race, gender, and household income. Methods. Data consisted of 6667 students that took part in the 1999 to 2006 National Health and Nutrition Examination Survey. The authors used adjusted Wald tests to examine differences in the prevalence of obesity (body mass index >95th percentile for age and sex) by sport for boys and girls separately. Results. Among adolescent youth age 12 to 19 years, 16.6% of male leisure time sport participants and 15.3% of female sport participants were obese, compared with 23.6% for male nonathlete participant-in-other-activities and 17.0% obesity rate for female nonathlete/participant-in-other-activities. For both males and females, reported participation in leisure time sports decreased between middle school and high school, and this reduction was associated with higher body mass index.


In vivo Mn-enhanced MRI for early tumor detection and growth rate analysis in a mouse medulloblastoma model.
Mouse models have increased our understanding of the pathogenesis of medulloblastoma (MB), the most common malignant pediatric brain tumor that often forms in the cerebellum. A major goal of ongoing research is to better understand the early stages of tumorigenesis and to establish the genetic and environmental changes that underlie MB initiation and growth. However, studies of MB progression in mouse models are difficult due to the heterogeneity of tumor onset times and growth patterns and the lack of clinical symptoms at early stages. Magnetic resonance imaging (MRI) is critical for noninvasive, longitudinal, three-dimensional (3D) brain tumor imaging in the clinic but is limited in resolution and sensitivity for imaging early MBs in mice. In this study, high-resolution (100 μm in 2 hours) and high-throughput (150 μm in 15 minutes) manganese-enhanced MRI (MEMRI) protocols were optimized for early detection and monitoring of MBs in a Patched-1 (Ptch1) conditional knockout (CKO) model. The high tissue contrast obtained with MEMRI revealed detailed cerebellar morphology and enabled detection of MBs over a wide range of stages including pretumoral lesions as early as 2 to 3 weeks postnatal with volumes close to 0.1 mm(3). Furthermore, longitudinal MEMRI allowed noninvasive monitoring of tumors and demonstrated that lesions within and between individuals have different tumorigenic potentials. 3D volumetric studies allowed quantitative analysis of MB tumor morphology and growth rates in individual Ptch1-CKO mice. These results show that MEMRI provides a powerful method for early in vivo detection and longitudinal imaging of MB progression in the mouse brain.


Universal DNA tag systems: a combinatorial design scheme.
Custom-designed DNA arrays offer the possibility of simultaneously monitoring thousands of hybridization reactions. These arrays show great potential for many medical and scientific applications, such as polymorphism analysis and genotyping. Relatively high costs are associated with the need to specifically design and synthesize problem-specific arrays. Recently, an alternative approach was suggested that utilizes fixed, universal arrays. This approach presents an interesting design problem-the arrays should contain as many probes as possible, while minimizing experimental errors caused by cross-hybridization. We use a simple thermodynamic model to cast this design problem in a formal mathematical framework. Employing new combinatorial ideas, we derive an efficient construction for the design problem and prove that our construction is near-optimal.