Cross-Sectional Analysis of the Relationship Between Home Blood Pressure and Indoor Temperature in Winter: A Nationwide Smart Wellness Housing Survey in Japan.
Mortality due to cardiovascular disease rises sharply in winter. Known as excess winter mortality, this phenomenon is partially explained by cold exposure-induced high blood pressure. Home blood pressure, especially in the morning, is closely associated with cardiovascular disease risk. We conducted the first large nationwide survey on home blood pressure and indoor temperature in 3775 participants (2095 households) who intended to conduct insulation retrofitting and were recruited by construction companies. Home blood pressure was measured twice in the morning and evening for 2 weeks. The relationship between home blood pressure and indoor temperature in winter was analyzed using a multilevel model with 3 levels: repeatedly measured day-level variables (eg, indoor ambient temperature and quality of sleep), nested within individual-level (eg, age and sex), and nested within household level. Cross-sectional analyses involving about 2900 participants (1840 households) showed that systolic blood pressure in the morning had significantly higher sensitivity to changes in indoor temperature (8.2 mm Hg increase/10°C decrease) than that in the evening (6.5 mm Hg increase/10°C decrease) in participants aged 57 years (mean age in this survey). We also found a nonlinear relationship between morning systolic blood pressure and indoor temperature, suggesting that the effect of indoor temperature on blood pressure varied depending on room temperature range. Interaction terms between age/women and indoor temperature were significant, indicating that systolic blood pressure in older residents and women was vulnerable to indoor temperature change. We expect that these results will be useful in determining optimum home temperature recommendations for men and women of each age group. Clinical Trial Registration- URL: http://www.umin.ac.jp/ctr/index.htm. Unique identifier: UMIN000030601.


Environmental health aspects of drinking water-borne outbreak due to karst flooding: case study.
Climate change may increase the incidence of waterborne diseases due to extreme rainfall events, and consequent microbiological contamination of the water source and supply. As a result of the complexity of the pathways from the surface to the consumer, it is difficult to detect an association between rainfall and human disease. The water supply of a Hungarian city, Miskolc (174,000 inhabitant), is mainly based on karstic water, a vulnerable underground water body. A large amount of precipitation fell on the catchment area of the karstic water source, causing an unusually strong karstic water flow and flooding, and subsequent microbiological contamination. The presence of several potential sources of contamination in the protective zone of the karstic water source should be emphasized. The water supplier was unprepared to treat the risk of waterborne outbreak caused by an extreme weather event. Public health intervention and hygienic measures were taken in line with epidemiological actions, focusing on the protection of consumers by providing safe drinking water. The contamination was identified, and measures were taken for risk reduction and prevention. This case study underlines the increasing importance of preparedness for extreme water events in order to protect the karstic water sources and to avoid waterborne outbreaks.


Soybean-derived Bowman-Birk Inhibitor (BBI) Inhibits HIV Replication in Macrophages.
The Bowman-Birk inhibitor (BBI), a soybean-derived protease inhibitor, is known to have anti-inflammatory effect in both in vitro and in vivo systems. Macrophages play a key role in inflammation and immune activation, which is implicated in HIV disease progression. Here, we investigated the effect of BBI on HIV infection of peripheral blood monocyte-derived macrophages. We demonstrated that BBI could potently inhibit HIV replication in macrophages without cytotoxicity. Investigation of the mechanism(s) of BBI action on HIV showed that BBI induced the expression of IFN-β and multiple IFN stimulated genes (ISGs), including Myxovirus resistance protein 2 (Mx2), 2',5'-oligoadenylate synthetase (OAS-1), Virus inhibitory protein (viperin), ISG15 and ISG56. BBI treatment of macrophages also increased the expression of several known HIV restriction factors, including APOBEC3F, APOBEC3G and tetherin. Furthermore, BBI enhanced the phosphorylation of IRF3, a key regulator of IFN-β. The inhibition of IFN-β pathway by the neutralization antibody to type I IFN receptor (Anti-IFNAR) abolished BBI-mediated induction of the anti-HIV factors and inhibition of HIV in macrophages. These findings that BBI could activate IFN-β-mediated signaling pathway, initialize the intracellular innate immunity in macrophages and potently inhibit HIV at multiple steps of viral replication cycle indicate the necessity to further investigate BBI as an alternative and cost-effective anti-HIV natural product.


UK national clinical audit: management of pregnancies in women with HIV.
The potential for HIV transmission between a pregnant woman and her unborn child was first recognized in 1982. Since then a complex package of measures to reduce risk has been developed. This project aims to review UK management of HIV in pregnancy as part of the British HIV Association (BHIVA) audit programme. The National Study of HIV in Pregnancy and Childhood (NSHPC), a population-based surveillance study, provided data for pregnancies with an expected delivery date from 1/1/13 - 30/6/14. Services also completed a survey on local management policies. Data were audited against the 2012 BHIVA pregnancy guidelines. During the audit period 1483 pregnancies were reported and 112 services completed the survey. Use of dedicated multidisciplinary teams was reported by 99% although 26% included neither a specialist midwife nor nurse. 17% of services reported delays >1 week for HIV specialist review of women diagnosed antenatally. Problematic urgent HIV testing had been experienced by 9% of services although in a further 49% the need for urgent testing had not arisen. Delays of >2 h in obtaining urgent results were common. Antiretroviral therapy (ART) was started during pregnancy in 37% women with >94% regimens in accordance with guidelines. Late ART initiation was common, particularly in those with a low CD4 count or high viral load. Eleven percent of services reported local policy contrary to guidelines regarding delivery mode for women with a VL <50 copies/mL at ≥36 weeks. According to NSHPC reports 27% of women virologically eligible for vaginal delivery planned to deliver by CS. Pregnant women in the UK are managed largely in accordance with BHIVA guidelines. Improvements are needed to ensure timely referral and ART initiation to ensure the best possible outcomes.


Community based study to compare the incidence and health services utilization pyramid for gastrointestinal, respiratory and dermal symptoms.
Gastrointestinal (GI), respiratory and dermal symptoms are common and cause substantial morbidity, although the information on their exact incidence and comparative burden is limited. The aim of this study was to describe the epidemiology and rate these three major symptom complexes in order to improve our understanding of the health burden imposed by these symptoms. We used data from a community based randomised control trial conducted from June 2007 to August 2008 among 277 South Australian families consuming rainwater. Using weekly health diaries, we prospectively collected information on GI (diarrhoea or vomiting), respiratory (sore throat, runny nose or cough) and dermal (rash, generalised itch or dermal infection) symptoms, as well as on relevant GP visits, time off work and/or hospitalisation due to these symptoms. Data were analysed using generalized estimating equations approach taking into account the variable number of weeks of follow-up of each individual and within-family clustering of responses. Over one year, at least one episode of GI symptoms was reported by 54% of participants (95% CI 50%-58%), at least one respiratory episode by 91% (95% CI 88%-93%) and at least one episode of dermal symptoms by 27% (95% CI 24%-30%). The average number of weeks per year during which respiratory symptoms occurred was four times greater than for GI or dermal symptoms (4.9, 1.2 and 1.2 weeks, respectively, p<0.001), with an average number of GP visits per person per year being twice as frequent (0.48, 0.26, 0.19 respectively, p<0.001). However, on a per episode basis, a higher proportion of people saw a GP or were hospitalised for GI symptoms. This first comparative study of three different symptom complexes showed that although respiratory symptoms are most common, GI symptoms cause a greater per episode burden on healthcare resources. Measuring and comparing the community based burden of these symptom complexes will assist evidence-based allocation of resources.


Spatial distribution of Raffaelea quercivora in xylem of naturally infested and inoculated oak trees.
Mass mortality of Japanese oak caused by Raffaelea quercivora due to Japanese oak wilt (JOW) has been tremendous since the late 1980s in Japan. We investigated detailed distribution of R. quercivora hyphae in a naturally infected Quercus serrata tree using fluorescein-conjugated wheat germ agglutinin and also examined spatial relationships between hyphal distribution, water conduction loss, and host reactions in xylem in inoculated Q. crispula saplings. Hyphae of R. quercivora elongated longitudinally in vessels and transversely in ray tissues in both naturally infected and inoculated Quercus trees. Hyphae were confined within a relatively small area near the inoculation site. Vessel dysfunction was also confined and overlapped with hyphal distribution. The reaction zone consisting of two types of fluorescent substance was formed surrounding the outside of the hyphal area in xylem and was always found in non-water-conductive zone. These results suggest that wilting of Quercus trees caused by JOW may not be induced by dysfunction of a small number of vessels, but by that of many vessels, and it requires that R. quercivora hyphae spread from many galleries bored by beetles during mass attacks.


Consistent condom use in South African youth's most recent sexual relationships.
Sexually active South African youth are at high risk for HIV infection but a low prevalence of condom use has been reported in this population. We examined correlates of consistent condom use with most recent sex partners among a nationally representative sample of youth 15-24 years old who reported having had sex in the previous 12 months (N = 6,649). Among men and women, having talked to a partner about using condoms was the most significant predictor of consistent condom use. However, youth who reported being in their most recent relationship for more than 1 year and who reported having had sex one or more times in the last month were more likely to report inconsistent condom use. HIV interventions should empower youth to talk about using condoms with their partners, encourage periodic testing for HIV, and reinforce condom use according to HIV status in long-term relationships.


How much toothpaste should a child under the age of 6 years use?
To discuss current concepts in the use of fluoride and to determine how much fluoride is sufficient for caries prevention but also how much is too much. Use of fluoride by young children is a balance between maximising caries efficacy and minimising the risk of fluorosis. Review of the current literature. This review considers the importance of amount, concentration and dose of fluoride applied from toothpaste and the implications for risk and benefit. Dental fluorosis is dependent on local fluoride levels in the extra cellular fluid surrounding the tooth during its development. These fluoride levels are determined by the plasma concentration that in turn is a function of the daily intake of fluoride. Fluoride released from bone during remodelling may also contribute to fluoride levels in the tissue. There is evidence to suggest that the effects of fluoride resulting in fluorosis prior to eruption of the tooth are cumulative and dependent on the amount and duration of exposure rather than a specific window of vulnerability. In contrast to dilution of ingested fluoride in the large volume of plasma, dilution of toothpaste in oral fluids is relatively small. Hence, for a given dose of fluoride, higher fluoride levels can be achieved in the oral environment using small amounts of toothpaste with higher fluoride concentrations rather than larger amounts with lower fluoride concentrations. It is concluded that for young children fluoride ingestion needs to be carefully controlled during the first six years of life and the best balance between risk and efficacy might be achieved by using small amounts of high fluoride toothpaste under close supervision from parents.


Beta-lactamase Enzymes of Clinical Pseudomonas aeruginosa Strains.
In this study, the production of extended spectrum beta-lactamase (ESBL), metallo-betalacatamase (MBL) and AmpC beta-lactamase enzymes of Pseudomonas aeruginosa (P aeruginosa) strains which were isolated from clinical samples were investigated. AmpC gene was also detected by the polymerase chain reaction (PCR) analysis. A hundred strains of P aeruginosa were included in the study. The presence of ESBL was investigated with combined disk confirmation test, MBL was investigated with E-test method and AmpC beta-lactamase was investigated with disk induction test. In order to detect the production of AmpC betalactamase genotypically, the PCR method was used. Only one strain was found to be MBL positive. Four per cent of strains were found to be ESBL positive. AmpC beta-lactamase production was positive in 73% of the strains with disk induction test. AmpC gene was detected in 96% of the studied strains with the PCR method. While ESBL and MBL rates in this study were significantly lower than those found in other studies, the rate of AmpC beta-lactamase was higher. Although AmpC gene was detected in some strains (23%), they were not found to produce AmpC beta-lactamase with disk induction test.


What Happens with the Circuit in Alzheimer's Disease in Mice and Humans?
A major mystery of many types of neurological and psychiatric disorders, such as Alzheimer's disease (AD), remains the underlying, disease-specific neuronal damage. Because of the strong interconnectivity of neurons in the brain, neuronal dysfunction necessarily disrupts neuronal circuits. In this article, we review evidence for the disruption of large-scale networks from imaging studies of humans and relate it to studies of cellular dysfunction in mouse models of AD. The emerging picture is that some forms of early network dysfunctions can be explained by excessively increased levels of neuronal activity. The notion of such neuronal hyperactivity receives strong support from in vivo and in vitro cellular imaging and electrophysiological recordings in the mouse, which provide mechanistic insights underlying the change in neuronal excitability. Overall, some key aspects of AD-related neuronal dysfunctions in humans and mice are strikingly similar and support the continuation of such a translational strategy.


Increasing Medical Student Interest in Nephrology.
Interest in nephrology careers is declining, possibly due to perceptions of the field and/or training aspects. Understanding practices of medical schools successfully instilling nephrology interest could inform efforts to attract leading candidates to the specialty. The American Society of Nephrology Workforce Committee's Best Practices Project was one of several initiatives to increase nephrology career interest. Board-certified nephrologists graduating medical school between 2002 and 2009 were identified in the American Medical Association Masterfile and their medical schools ranked by production. Renal educators from the top 10 producing institutions participated in directed focus groups inquiring about key factors in creating nephrology career interest, including aspects of their renal courses, clinical rotations, research activities, and faculty interactions. Thematic content analysis of the transcripts (with inductive reasoning implementing grounded theory) was performed to identify factors contributing to their programs' success. The 10 schools identified were geographically representative, with similar proportions of graduates choosing internal medicine (mean 26%) as the national graduating class (26% in the 2017 residency Match). Eighteen educators from 9 of these 10 institutions participated. Four major themes were identified contributing to these schools' success: (1) nephrology faculty interaction with medical students; (2) clinical exposure to nephrology and clinical relevance of renal pathophysiology materials; (3) use of novel educational modalities; and (4) exposure, in particular early exposure, to the breadth of nephrology practice. Early and consistent exposure to a range of clinical nephrology experiences and nephrology faculty contact with medical students are important to help generate interest in the specialty.


Proton polarization enhancement of up to 150 with dynamic nuclear polarization of plasma-treated glucose powder.
Dynamic nuclear polarization (DNP) for the enhancement of the NMR signals of specific metabolites has recently found applications in the context of magnetic resonance imaging (MRI). Currently, DNP signal enhancement is implemented in clinical systems through the use of exogenous stable organic free radicals, known as polarization agents (PAs), mixed in a solution with the metabolite of interest. These PAs are medically undesirable and thus must be filtered out prior to patient injection - a task that involves considerable technical complexity and consumes valuable time during which the polarization decays. Here, we aim to demonstrate DNP enhancements large enough for clinical relevance using a process free of exogenous PAs. This is achieved by processing (soft grinding) the metabolite in its solid form and subsequently exposing it to plasma in a dilute atmosphere to produce chemically-unstable free radicals (herein referred to as electrical-discharge-induced radicals - EDIRs) within the powder. These samples are then subjected to the normal DNP procedure of microwave irradiation while placed under a high static magnetic field, and their NMR signal is measured to quantify the enhancement of the protons' signal in the solid. Proton signal enhancements (measured as the ratio of the NMR signal with microwave irradiation to the NMR signal without microwave irradiation) of up to 150 are demonstrated in glucose. Upon fast dissolution, the free radicals are annihilated, leaving the sample in its original chemical composition (which is safe for clinical use) without any need for filtration and cumbersome quality control procedures. We thus conclude that EDIRs are found to be highly efficient in providing DNP enhancement levels that are on par with those achieved with the exogenous PAs, while being safe for clinical use. This opens up the possibility of applying our method to clinical scenarios with minimal risks and lower costs per procedure.


Development and validation of a HPLC method for the determination of benzo(a)pyrene in human breast milk.
A simple analytical procedure was developed for the quantitation of benzo(a)pyrene in human breast milk using solid phase extraction (SPE) combined with high performance liquid chromatography. Before the chromatographic process, SPE, including C18 functional groups in silicagel cartridges, was conducted for sample preparation. A C18 column (100×4.6 mm id, 3 μm particle size) was used with acetonitrile:water (80:20) as the mobile phase at a flow rate 1mL/min at 30°C. Fluorimetric detection was performed for excitation and emission at 290 and 406 nm, respectively. It was observed that the calibration curve was linear over the range of 0.5-80 ng/mL. The limit of detection and limit of quantitation were found to be 0.5 and 1.07 ng/mL, respectively. Intraday and interday relative standard deviation values were less than 5.15%. Moreover, the newly developed method provides a fast, simple, cost effective, and sensitive assay to detect an important carcinogen substance, benzo(a)pyrene, in human breast milk.


Antitumor activity mediated by CpG: the route of administration is critical.
Unmethylated CpG oligodeoxynucleotides (CpG) are synthetic toll-like receptor 9 agonists that activate innate immune cells and which have been tested as an immune therapy in a number of cancer clinical trials. Although some antitumor immune responses have been reported, so far the majority of studies have failed to show significant clinical responses to CpG. Here we showed that the route of administration is critical to the antitumor activity of CpG. Although intravenous (i.v.) injection of CpG was capable of inducing the activation and expansion of tumor antigen-specific T cells, most of these activated T cells failed to migrate to tumor sites. By contrast, intratumoral (i.t.) injection of CpG led to extensive tumor infiltration of antigen-specific T cells and subsequent tumor suppression. We further showed that very high levels of inflammatory chemokines [regulated upon activation, normal T-cell expressed, and secreted (RANTES), interferon-inducible protein-10 (IP-10), monocyte chemoattractant protein-1, monocyte chemotactic protein (MCP5), macrophage inflammatory proteins (MIP1α, and MIP1β)] were induced in the tumor microenvironment after i.t. CpG injection, compared with administration by the i.v. route. It is interesting to note that, in vivo depletion of plasmacytoid dendritic cells greatly reduced the levels of chemokines induced; also, T-cell accumulation and antitumor effect were impaired. We also showed that i.t. but not i.v. CpG injection induced a broad antigen-specific T-cell response against tumor-derived antigens. Collectively, our data provides evidence that the route of CpG administration is a critical factor in mediating antitumor activity. By inducing localized inflammatory signals at tumor sites, i.t. CpG effectively promotes the migration, activation and function of immune cells, ultimately leading to improved tumor control.


Beyond Classroom Academics: A School-Wide and Multi-Contextual Perspective on Student Engagement in School.
School engagement researchers have historically focused on academic engagement or academic-related activities. Although academic engagement is vital to adolescents' educational success, school is a complex developmental context in which adolescents also engage in social interactions while exploring their interests and developing competencies. In this article, school engagement is re-conceptualized as a multi-contextual construct that includes both academic and social contexts of school. The authors begin by describing how the characteristics of these contexts provide the opportunities and resources for adolescents to engage in academic learning and social interactions throughout school. Motivational theories are then used as an operational framework for understanding how adolescents become engaged in school, which is followed by a discussion about how adolescents' academic and social engagement interact to shape their academic achievement. The article concludes with implications for practice and future research.


[Case finding in family practice and neuropsychological diagnosis at a memory clinic].
To improve early diagnosis of cognitive impairment in the elderly a case-finding strategy should be adopted: general practitioners should only examine "suspicious" patients with a very efficient tool. The BrainCheck, a new case-finding tool from Switzerland, comprises of three questions for the patient and the clock drawing test. In addition the family member fills out a short questionnaire. In a validation study with 113 patients and 70 cognitively healthy individuals the BrainCheck showed a correct classification rate of 89 % (sensitivity = 97 %; specificity = 82 %). Within the work of a memory clinic the neuropsychological assessment should use appropriately normed cognitive instruments, especially if one is to detect subtle cognitive impairment. DSM-5 proposes to replace the terms mild cognitive impairment and dementia with mild and severe neurocognitive disorder, respectively. A major improvement in DSM-5 is the list of cognitive domains, which need to be examined. However, no standardized or widely accepted tool exists today to assess the domain of social cognition.


The place of medicine in the American prison: ethical issues in the treatment of offenders.
In Britain doctors and others concerned with the treatment of offenders in prison may consult the Butler Report (see Focus, pp 157) and specialist journals, but these sources are concerned with the system in Britain only. In America the situation is different, both in organization and in certain attitudes. Dr Peter L Sissons has therefore provided a companion article to that of Dr Paul Bowden (page 163) describing the various medical issues in prisons. The main difference between the treatment of offenders in prisons in America and in Britain lies in the nature of the federal system which means that each state may operate a different system in a variety of prisons and prison medical services are as various. Nationally, the prison systems are 'structured to treat and cure the offender'. Therefore it follows that the prison medical officer is only one of the professionals concerned with this 'cure' of the offender. This principle also applies to any form of research: medical research in prisons is part of a programme which covers a wide field of social and judicial research. The prison medical officer (where there is one) has of course to look after sick prisoners, and the American idea of 'cure' is also expressed in the need for more corrective surgery where, for example, it is necessary to remove physical impediments to social rehabilitation. But a doctor is only found on the staff of those institutions which are large: in the smaller prisons there may be only first-aid facilities, and no specially appointed doctor in the community. Moreover medicines are often dispensed by medical auxiliaries who are sometimes prisoners themselves. Finally, in America prisoners are regularly invited to volunteer as subjects for medical and social research for which they are paid. In short, although it is hoped to 'cure' a prisoner he is a criminal first and a patient second.


Evaluation of bone healing with eggshell-derived bone graft substitutes in rat calvaria: a pilot study.
The purpose of this study was to investigate the potential effectiveness of a surface-modified natural calcium carbonate, hen eggshell (ES) as a bone graft substitute. The surface characteristics, cell viability on, and osteoconductivity of, particulated ES with and without hydrothermal treatment in phosphate solutions were evaluated. Hydrothermal treatment partially converted ES to calcium-deficient hydroxyapatite (HA) with surface microstructure. MTT assay indicated higher osteoblast viability on surface-modified ES compared with a commercially available bone substitute, anorganic bovine bone (Bio-Oss, BO) (p < 0.001). Histological and histomorphometric analysis showed significantly greater new bone formation and mineralized bone-to-graft contact of surface-modified ES, especially with hydrothermally treated ES, compared with BO in 5-mm diameter calvarial defects in rats at 4 and 8 weeks of healing (p < 0.01). Complete bony bridging was more frequently found with hydrothermally treated ES. The results of this pilot study indicate the potential efficacy of surface-modified particulated hen eggshell as an osteoconductive bone substitute in a rat calvarial defect model.


Behavioral and Neuropathological Changes After Toxoplasma gondii Ocular Conjunctival Infection in BALB/c Mice.
Ocular infection with Toxoplasma gondii causes toxoplasmosis in mice. However, following ocular infection with tachyzoites, the cause of the accompanying progressive changes in hippocampal-dependent tasks, and their relationship with the morphology and number of microglia, is less well understood. Here, in 6-month-old, female BALB/c mice, 5 μl of a suspension containing 48.5 × 106 tachyzoites/ml was introduced into the conjunctival sac; control received an equal volume of saline. Before and after instillation, all mice were subject to an olfactory discrimination (OD) test, using predator (cat) feces, and to an open-field (OF) task. After the behavioral tests, the animals were culled at either 22 or 44 days post-instillation (dpi), and the brains and retinas were dissected and processed for immunohistochemistry. The total number of Iba-1-immunolabeled microglia in the molecular layer of the dentate gyrus was estimated, and three-dimensional reconstructions of the cells were evaluated. Immobility was increased in the infected group at 12, 22, and 43 dpi, but the greatest immobility was observed at 22 dpi and was associated with reduced line crossing in the OF and distance traveled. In the OD test, infected animals spent more time in the compartment with feline fecal material at 14 and at 43 dpi. No OD changes were observed in the control group. The number of microglia was increased at 22 dpi but returned to control levels by 44 dpi. These changes were associated with the differentiation of T. gondii tachyzoites into bradyzoite-enclosed cysts within the brain and retina. Thus, infection of mice with T. gondii alters exploratory behavior, gives rise to a loss in predator's odor avoidance from 2 weeks after infection, increased microglia number, and altered their morphology in the molecular layer of the dentate gyrus.


Lysophosphatidic acid increases the proliferation and migration of adipose‑derived stem cells via the generation of reactive oxygen species.
Phospholipid derivatives, such as lysophosphatidic acid (LPA), exhibit mitogenic effects on mesenchymal stem cells; however, the molecular mechanism underlying this stimulation has yet to be identified. The aims of the present study were as follows: To evaluate the stimulatory effects of LPA on the proliferation and migration of adipose‑derived stem cells (ASCs); to study the association between reactive oxygen species (ROS) and LPA signaling in ASCs; and to investigate the microRNAs upregulated by LPA treatment in ASCs. The results of the present study demonstrated that LPA increased the proliferation and migration of ASCs, and acted as a mitogenic signal via extracellular signal‑regulated kinases 1/2 and the phosphoinositide 3‑kinase/Akt signaling pathways. The LPA1 receptor is highly expressed in ASCs, and pharmacological inhibition of it by Ki16425 significantly attenuated the proliferation and migration of ASCs. In addition, LPA treatment generated ROS via NADPH oxidase 4, and ROS were able to function as signaling molecules to increase the proliferation and migration of ASCs. The induction of ROS by LPA treatment also upregulated the expression of miR‑210. A polymerase chain reaction array assay demonstrated that the expression levels of adrenomedullin and Serpine1 were increased following treatment with LPA. Furthermore, transfection with Serpine1‑specific small interfering RNA attenuated the migration of ASCs. In conclusion, the present study is the first, to the best of our knowledge, to report that ROS generation and miR‑210 expression are associated with the LPA‑induced stimulation of ASCs, and that Serpine1 mediates the LPA‑induced migration of ASCs. These results further suggest that LPA may be used for ASC stimulation during stem cell expansion.


A family of rhodium and iridium complexes with semirigid benzylsilyl phosphines: from bidentate to tetradentate coordination modes.
The synthesis of a new trisbenzylsilanephosphine P{(o-C6H4CH2)SiMe2H}3 (1) is shown to proceed with high yields from P(o-tolyl)3. Compound 1 coordinates to the Rh and Ir dimers [MCl(COD)]2 (M = Rh, Ir) in a tetradentate or tridentate fashion, depending on the strict exclusion of water. The dimeric compounds [ClM(SiMe2CH2-o-C6H4)2P(o-C6H4-CH2SiMe2H)]2, 2Rh and 2Ir, feature a tetradentate coordination of the starting ligand with P and two Si atoms as well as a non-classical agostic Si-H group. The presence of adventitious water in the solvents leads to the formation of two new complexes [(μ2-Cl)2M2(SiMe2CH2-o-C6H4)2P(o-C6H4-CH2SiMe2OSiMe2CH2-o-C6H4-)P(SiMe2CH2-o-C6H4)2], 3Rh and 3Ir, which feature a siloxane bridge through Si-H bond breaking in 2. Reaction of [RhCl(COD)]2 with the bisbenzylsilanephosphine PhP{(o-C6H4CH2)SiMe2H}2 leads to the formation of compound 4Rh which features also a dimeric structure with the SiPSi ligand coordinated through the two silicon atoms, one of which occupies the apical position of a square-pyramidal geometry in the solid state, while the second is disposed equatorially trans to π-donor Cl. Finally, bidentate coordination of a PSi ligand is achieved by reaction of [RhCl(COD)]2 with Ph2P{(o-C6H4CH2)SiMe2H} which leads to the monometallic species [RhCl(SiMe2CH2-o-C6H4-PPh2)2], 5Rh, incorporating two chelating PSi ligands and maintaining a Cl ligand.


Long-Term Warming in Alaska Enlarges the Diazotrophic Community in Deep Soils.
Tundra ecosystems are typically carbon (C) rich but nitrogen (N) limited. Since biological N2 fixation is the major source of biologically available N, the soil N2-fixing (i.e., diazotrophic) community serves as an essential N supplier to the tundra ecosystem. Recent climate warming has induced deeper permafrost thaw and adversely affected C sequestration, which is modulated by N availability. Therefore, it is crucial to examine the responses of diazotrophic communities to warming across the depths of tundra soils. Herein, we carried out one of the deepest sequencing efforts of nitrogenase gene (nifH) to investigate how 5 years of experimental winter warming affects Alaskan soil diazotrophic community composition and abundance spanning both the organic and mineral layers. Although soil depth had a stronger influence on diazotrophic community composition than warming, warming significantly (P < 0.05) enhanced diazotrophic abundance by 86.3% and aboveground plant biomass by 25.2%. Diazotrophic composition in the middle and lower organic layers, detected by nifH sequencing and a microarray-based tool (GeoChip), was markedly altered, with an increase of α-diversity. Changes in diazotrophic abundance and composition significantly correlated with soil moisture, soil thaw duration, and plant biomass, as shown by structural equation modeling analyses. Therefore, more abundant diazotrophic communities induced by warming may potentially serve as an important mechanism for supplementing biologically available N in this tundra ecosystem.IMPORTANCE With the likelihood that changes in global climate will adversely affect the soil C reservoir in the northern circumpolar permafrost zone, an understanding of the potential role of diazotrophic communities in enhancing biological N2 fixation, which constrains both plant production and microbial decomposition in tundra soils, is important in elucidating the responses of soil microbial communities to global climate change. A recent study showed that the composition of the diazotrophic community in a tundra soil exhibited no change under a short-term (1.5-year) winter warming experiment. However, it remains crucial to examine whether the lack of diazotrophic community responses to warming is persistent over a longer time period as a possibly important mechanism in stabilizing tundra soil C. Through a detailed characterization of the effects of winter warming on diazotrophic communities, we showed that a long-term (5-year) winter warming substantially enhanced diazotrophic abundance and altered community composition, though soil depth had a stronger influence on diazotrophic community composition than warming. These changes were best explained by changes in soil moisture, soil thaw duration, and plant biomass. These results provide crucial insights into the potential factors that may impact future C and N availability in tundra regions.


Internal dispersal of seeds by waterfowl: effect of seed size on gut passage time and germination patterns.
Long distance dispersal may have important consequences for gene flow and community structure. The dispersal of many plants depends on transport by vertebrate seed dispersers. The shapes of seed shadows produced by vertebrates depend both on movement patterns of the dispersers and on the dynamics and effects of passage through the disperser's gut (i.e. the retention time, survival and germination of ingested seeds). A combination of experiments with captive waterbirds and aquatic plant seeds was used to analyse the following: (a) the effects of inter- and intra-specific variation in seed size and duck species on seed retention time in the gut and (b) the relationship between retention time and the percent germination and germination rates of seeds. Among the three Scirpus species used, those with smaller seeds showed higher survival after ingestion by birds and longer retention times inside their guts than those with larger seeds. For Potamogeton pectinatus, only seeds from the smaller size class (<8 mg) survived ingestion. Retention time affected the percent germination and germination rate of Scirpus seeds but in a manner that varied for the different plant and bird species studied. We recorded both linear and non-linear effects of retention time on percent germination. In addition, germination rate was positively correlated with retention time in Scirpus litoralis but negatively correlated in Scirpus lacustris. Small seed size can favour dispersal over larger distances. However, the effects of retention time on percent germination can modify the seed shadows produced by birds due to higher percent germination of seeds retained for short or intermediate periods. The changes in dispersal quality associated with dispersal distance (which is expected to be positively related to retention time) will affect the probability of seedling establishment over longer distances and, thus, the spatial characteristics of the effective seed shadow.


Co-administration of vismodegib with rosiglitazone or combined oral contraceptive in patients with locally advanced or metastatic solid tumors: a pharmacokinetic assessment of drug-drug interaction potential.
Vismodegib, a first-in-class oral hedgehog pathway inhibitor, is an effective treatment for advanced basal cell carcinoma. Based on in vitro data, a clinical drug-drug interaction (DDI) assessment of cytochrome P450 (CYP) 2C8 was necessary; vismodegib's teratogenic potential warranted a DDI study with oral contraceptives (OCs). This single-arm, open-label study included two cohorts of patients with locally advanced or metastatic solid malignancies [Cohort 1: rosiglitazone 4 mg (selective CYP2C8 probe); Cohort 2: OC (norethindrone 1 mg/ethinyl estradiol 35 μg; CYP3A4 substrate)]. On Day 1, patients received rosiglitazone or OC. On Days 2-7, patients received vismodegib 150 mg/day. On Day 8, patients received vismodegib plus rosiglitazone or OC. The effect of vismodegib on rosiglitazone and OC pharmacokinetic parameters (primary objective) was evaluated through pharmacokinetic sampling over a 24-h period (Days 1 and 8). The mean ± SD vismodegib steady-state plasma concentration (Day 8, N = 51) was 20.6 ± 9.72 μM (range 7.93-62.4 μM). Rosiglitazone AUC(0-inf) and C(max) were similar with concomitant vismodegib [≤8% change in geometric mean ratios (GMRs); N = 24]. Concomitant vismodegib with OC did not affect ethinyl estradiol AUC(0-inf) and C(max) (≤5% change in GMRs; N = 27); norethindrone C(max) and AUC(0-inf) GMRs were higher (12 and 23%, respectively) with concomitant vismodegib. This DDI study in patients with cancer demonstrated that systemic exposure of rosiglitazone (a CYP2C8 substrate) or OC (ethinyl estradiol/norethindrone) is not altered with concomitant vismodegib. Overall, there appears to be a low potential for DDIs when vismodegib is co-administered with other medications.


The preventive effect of magnesium on coronary spasm in patients with vasospastic angina.
Previous studies have reported that magnesium (Mg) deficiency is associated with coronary spasm. However, little is known about the preventive effect of Mg on coronary spasm. The present study investigated whether Mg prevents coronary spasm in patients with vasospastic angina (VSA). Effectiveness trial. University medical center. Twenty-two patients with VSA. Coronary spasm was induced with an intracoronary infusion of acetylcholine (Ach). After spontaneous relief of the coronary spasm, Mg sulfate (0.27 mmol/kg body weight) was infused IV over 20 min in 14 patients and isotonic glucose was infused in 8 patients as control subjects. Intracoronary infusion of Ach was then repeated, and the diameter of the coronary arteries was measured quantitatively. Mg infusion caused coronary artery dilatation at baseline in both the spastic (5. 9 +/- 2.3%) and nonspastic segments (5.5 +/- 1.5%). Mg infusion reduced the severity of chest pain and ST-segment deviations during coronary spasm. After the Mg infusion, the percent change in the diameter of the spastic segments improved from - 62.8 +/- 2.6% to - 43.7 +/- 4.7% during coronary spasm. Overall, 10 of 14 patients (71%) responded favorably to Mg infusion. Isotonic glucose infusion did not elicit changes in chest pain severity, ST-segment deviations, or the diameter of the coronary arteries during spasm. Mg infusion produces nonsite-specific basal coronary dilatation and suppresses Ach-induced coronary spasm in patients with VSA.


Recoil-proton polarization in high-energy deuteron photodisintegration with circularly polarized photons.
We measured the angular dependence of the three recoil-proton polarization components in two-body photodisintegration of the deuteron at a photon energy of 2 GeV. These new data provide a benchmark for calculations based on quantum chromodynamics. Two of the five existing models have made predictions of polarization observables. Both explain the longitudinal polarization transfer satisfactorily. Transverse polarizations are not well described, but suggest isovector dominance.


trans-Lesion synthesis past bulky benzo[a]pyrene diol epoxide N2-dG and N6-dA lesions catalyzed by DNA bypass polymerases.
The effectiveness of in vitro primer elongation reactions catalyzed by human bypass DNA polymerases kappa (hDinB1), pol eta (hRad30A), pol iota (hRad30B), and yeast pol zeta (Rev3 and Rev7) in site-specifically modified template oligonucleotide strands were studied in vitro. The templates contained single bulky lesions derived from the trans-addition of the mutagenic (+)- or (-)-enantiomers of r7,t8-dihydroxy-t9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (a metabolite of the environmental carcinogen benzo[a]pyrene), to the exocyclic amino groups of guanine or adenine in oligonucleotide templates 33, or more, bases long. In "running start" primer extension reactions, pol kappa effectively bypassed both the stereoisomeric (+)- and (-)-trans-guanine adducts but not the analogous adenine adducts. In sharp contrast, pol eta, which exhibits considerable sequence homology with pol kappa (both belong to the group of Y family polymerases), is partially blocked by the guanine adducts and the (-)-trans-adenine adduct, although the stereoisomeric (+)-trans-adenine adduct is more successfully bypassed. Neither pol iota nor pol zeta, either alone or in combination, were effective in trans-lesion synthesis past the same adducts. In all cases, the fidelity of insertion is dependent on adduct stereochemistry and structure. Generally, error-free nucleotide insertion opposite the lesions tends to depend more on adduct stereochemistry than error-prone insertion. None of the polymerases tested are a universal bypass polymerase for the stereoisomeric bulky polycyclic aromatic hydrocarbon-DNA adducts derived from anti-BPDE.


Pregnancy-related school dropout and prior school performance in KwaZulu-Natal, South Africa.
This study uses data from KwaZulu-Natal, South Africa, to examine the factors associated with schoolgirl pregnancy and subsequent educational pathways. We find that prior schooling discontinuities--defined as instances of non-pregnancy-related grade repetition or temporary withdrawals from school--are strongly associated with a young woman's likelihood of later becoming pregnant while enrolled in school, dropping out of school if she becomes pregnant, and not returning to school following a pregnancy-related dropout. Young women who are the primary caregivers to their children are also significantly more likely to leave school than are those who have help with their childcare responsibilities. Given the increasing importance of female school participation in sub-Saharan Africa, programs must be designed to reach girls and their families early to increase incentives for ensuring their timely progression through school and to increase their access to reproductive health information and services.


Leisure activities, education, and cognitive impairment in Chinese older adults: a population-based longitudinal study.
We examine the association between leisure-time activities and the risk of developing cognitive impairment among Chinese older people, and further investigate whether the association varies by educational level. This follow-up study included 6,586 participants (aged 79.5 ± 9.8 years, range 65-105 years, 51.7% female) of the Chinese Longitudinal Healthy Longevity Survey who were aged ≥65 years and were free of cognitive impairment in 2002. Incident cognitive impairment was defined at the 2005 or 2008/2009 survey following an education-based cut-off on the adapted Chinese version of Mini-Mental State Examination (MMSE). Participation in cognitive activities (e.g. reading) and non-exercise physical activity (e.g. housework) was assessed by a self-reported scale. Cox proportional hazard models were employed to examine the association of leisure activities with incident cognitive impairment while controlling for age, gender, education, occupation, residence, physical exercise, smoking, drinking, cardiovascular diseases and risk factors, negative well-being, and physical functioning, and baseline MMSE score. During a five-year follow-up, 1,448 participants developed incident cognitive impairment. Overall, a high level of participation in leisure activities was associated with a 41% decreased risk of cognitive impairment compared to low-level engagement in leisure activities after controlling for age, gender, education, and other confounders. Moreover, there was a significant interaction between leisure activity and educational level, such that the beneficial effect of leisure activities on cognitive function was larger in educated elderly than their uneducated counterparts, and only educated elderly benefited from cognitive activities. Late-life leisure activities protect against cognitive impairment among elderly Chinese people, and the protective effects are more profound for educated elderly.


Transcription, nucleosome positioning and protein binding modulate nucleotide excision repair of the Saccharomyces cerevisiae MET17 promoter.
We have assessed how transcription, chromatin structure and protein binding modulate nucleotide excision repair in the upstream regulatory region and early coding region of the endogenous Saccharomyces cerevisiae gene MET17. Removal of UV-induced cyclobutane pyrimidine dimers was measured from these regions, in which transcription and chromatin structure could be regulated independently of each other. Distinct repair trends were apparent depending on transcriptional state. When transcription was repressed nucleosome positioning and protein binding as determined by chromatin immunoprecipitation and quantitative real-time PCR, were significant factors. Nucleosome positioning and/or protein binding effects were most apparent on the strand that becomes transcribed, with repair occurring fastest in a nucleosome free region but being retarded where regulatory proteins bound within this region. When transcription was derepressed the rate of repair increased on both strands in a region beginning 200 bp upstream of the TATA box and extending downstream into the coding region. This effect overrode the influences of nucleosome positioning and protein binding.


A comparative study of American and Chinese college students' motives for food choice.
Previous cross-cultural research has examined college students' food choice decisions in different countries. The current study aimed to add to the literature by examining similarities and differences in motives for food choice between American (N = 328) and Chinese (N = 333) college students. The Food Choice Questionnaire (FCQ) was used to measure the participants' motives for food choice. Students' perceptions on the importance of diet and on their body satisfaction were also obtained. Results show that, while there are many similarities between the two cultural groups on the FCQ items, there are also significant differences. Specifically, the two groups view sensory appeal, weight, health, mood, and familiarity in a similar way, but the American participants score higher on price and convenience whereas the Chinese score higher on natural content and ethical concerns. We believe contextual cultural factors of each country may be related to these results. Women view sensory appeal and weight as significantly more important than men. Interactions between culture and gender are also found. For example, American women score significantly higher than American men on mood whereas there is no gender difference in the Chinese group; on the other hand, Chinese men score significantly higher on price than Chinese women whereas there is no gender difference in the American group.


Identifying gravitational insecurity in children: a pilot study.
This study developed an observational assessment of gravitational insecurity (GI), the GI Assessment, and examined its preliminary reliability and validity evidence. The GI Assessment consisted of 15 activities that created conditions characterized as fear-inducing for children with gravitational insecurity. Three behavioral categories-avoidance, emotional, and postural responses--were scored for each activity. Participants were 18 children with gravitational insecurity, ages 5-10 years, and a matched group of children who were typically developing. Forty-eight preschoolers who were typically developing, ages 2-4 years, were examined for developmental trends. Significant differences were found between groups. Discriminant analysis classified 83% of the gravitationally insecure group and 100% of the typical group. Interrater reliability for the total test was .79. Performance of preschool children suggested a developmental age trend. The GI Assessment is a promising clinical tool for objectively identifying children with gravitational insecurity.


The testate lobose amoebae (order Arcellinida Kent, 1880) finally find their home within Amoebozoa.
Testate lobose amoebae (order Arcellinida Kent, 1880) are common in all aquatic and terrestrial habitats, yet they are one of the last higher taxa of unicellular eukaryotes that has not found its place in the tree of life. The morphological approach did not allow to ascertain the evolutionary origin of the group or to prove its monophyly. To solve these challenging problems, we analyzed partial small-subunit ribosomal RNA (SSU rRNA) genes of seven testate lobose amoebae from two out of the three suborders and seven out of the 13 families belonging to the Arcellinida. Our data support the monophyly of the order and clearly establish its position among Amoebozoa, as a sister-group to the clade comprising families Amoebidae and Hartmannellidae. Complete SSU rRNA gene sequences from two species and a partial actin sequence from one species confirm this position. Our phylogenetic analyses including representatives of all sequenced lineages of lobose amoebae suggest that a rigid test appeared only once during the evolution of the Amoebozoa, and allow reinterpretation of some morphological characters used in the systematics of Arcellinida.


Malo-ethanolic fermentation in Saccharomyces and Schizosaccharomyces.
Yeast species are divided into the K(+) or K(-) groups, based on their ability or inability to metabolise tricarboxylic acid (TCA) cycle intermediates as sole carbon or energy source. The K(-) group of yeasts includes strains of Saccharomyces, Schizosaccharomyces pombe and Zygosaccharomyces bailii, which is capable of utilising TCA cycle intermediates only in the presence of glucose or other assimilable carbon sources. Although grouped together, these yeasts have significant differences in their abilities to degrade malic acid. Typically, strains of Saccharomyces are regarded as inefficient metabolisers of extracellular malic acid, whereas strains of Sch. pombe and Z. bailii can effectively degrade high concentrations of malic acid. The ability of a yeast strain to degrade extracellular malic acid is dependent on both the efficient transport of the dicarboxylic acid and the efficacy of the intracellular malic enzyme. The malic enzyme converts malic acid into pyruvic acid, which is further metabolised to ethanol and carbon dioxide under fermentative conditions via the so-called malo-ethanolic (ME) pathway. This review focuses on the enzymes involved in the ME pathway in Sch. pombe and Saccharomyces species, with specific emphasis on the malate transporter and the intracellular malic enzyme.


Nonlinear spike-and-slab sparse coding for interpretable image encoding.
Sparse coding is a popular approach to model natural images but has faced two main challenges: modelling low-level image components (such as edge-like structures and their occlusions) and modelling varying pixel intensities. Traditionally, images are modelled as a sparse linear superposition of dictionary elements, where the probabilistic view of this problem is that the coefficients follow a Laplace or Cauchy prior distribution. We propose a novel model that instead uses a spike-and-slab prior and nonlinear combination of components. With the prior, our model can easily represent exact zeros for e.g. the absence of an image component, such as an edge, and a distribution over non-zero pixel intensities. With the nonlinearity (the nonlinear max combination rule), the idea is to target occlusions; dictionary elements correspond to image components that can occlude each other. There are major consequences of the model assumptions made by both (non)linear approaches, thus the main goal of this paper is to isolate and highlight differences between them. Parameter optimization is analytically and computationally intractable in our model, thus as a main contribution we design an exact Gibbs sampler for efficient inference which we can apply to higher dimensional data using latent variable preselection. Results on natural and artificial occlusion-rich data with controlled forms of sparse structure show that our model can extract a sparse set of edge-like components that closely match the generating process, which we refer to as interpretable components. Furthermore, the sparseness of the solution closely follows the ground-truth number of components/edges in the images. The linear model did not learn such edge-like components with any level of sparsity. This suggests that our model can adaptively well-approximate and characterize the meaningful generation process.


Importance of positioning for microbial evolution.
Microbes commonly live in dense surface-attached communities where cells layer on top of one another such that only those at the edges have unimpeded access to limiting nutrients and space. Theory predicts that this simple spatial effect, akin to plants competing for light in a forest, generates strong natural selection on microbial phenotypes. However, we require direct empirical tests of the importance of this spatial structuring. Here we show that spontaneous mutants repeatedly arise, push their way to the surface, and dominate colonies of the bacterium Pseudomonas fluorescens Pf0-1. Microscopy and modeling suggests that these mutants use secretions to expand and push themselves up to the growth surface to gain the best access to oxygen. Physically mixing the cells in the colony, or introducing space limitations, largely removes the mutant's advantage, showing a key link between fitness and the ability of the cells to position themselves in the colony. We next follow over 500 independent adaptation events and show that all occur through mutation of a single repressor of secretions, RsmE, but that the mutants differ in competitiveness. This process allows us to map the genetic basis of their adaptation at high molecular resolution and we show how evolutionary competitiveness is explained by the specific effects of each mutation. By combining population level and molecular analyses, we demonstrate how living in dense microbial communities can generate strong natural selection to reach the growing edge.


Can we control the electronic energy transfer in molecular dyads through metal nanoparticles? A QM/continuum investigation.
Metal nanoparticles (MNPs) can have a dramatic effect on the electronic energy transfer (EET) between donor and acceptor molecular dyes. In addition, such an effect can be modulated by the presence of a solvent. Here we present a novel multiscale QM/continuum approach which can both treat MNP-solvent-mediated EET and take into account the effects of the MNPs in all the photophysical processes into play (absorption and emission), including the competitive energy transfer from the molecular systems to the metal. By applying such a unified theoretical framework, we show that the excitonic interactions in stacked dimers are generally reduced by the presence of MNPs. In contrast, for setups in which the two transferring moieties are separated by the MNPs, the presence of the metal results in a direct enhancement of the coupling but, when the competing process of quenching by the MNP is also considered, the final effect is almost invariably a reduction of the efficiency of the EET process. Only for particular donor-MNP-acceptor setups, the model shows that waveguide-like behavior can be obtained, in these cases the excitation energy of the donor can be transferred to the acceptor over distances much longer than those allowed by the conventional Förster mechanism.


Differences in patterns of meningoencephalitis due to bacterial kidney disease in farmed Atlantic and chinook salmon.
A range of neural pathology, in particular meningitis with sporadic encephalitic extension, can develop in salmonids infected with Renibacterium salmoninarum, the agent of bacterial kidney disease (BKD). Tissues from 134 Atlantic salmon and 164 chinook salmon were selected on the basis of their having a positive diagnosis of BKD, with evidence of multi-tissue infection including the brain (214 fish), or brain involvement in the absence of systemic lesions attributable to BKD (58 fish). Although meningitis was a feature of BKD in both species, encephalitis was more common in the Atlantic salmon. Specifically, a higher portion of the Atlantic salmon had encephalitis accompanying meningitis (P = 0.0159), encephalitis in the absence of meningitis (P = 0.0756), and brain lesions (meningitis or encephalitis) in the apparent absence of systemic lesions (P = 0.0067). These findings suggest either that some aspects of the pathobiology of R salmoninarum are dictated by the host species, or that the farm management methods used to deal with BKD are sufficiently different for the two species of salmon that they affect the pathology of the disease.


Multi-platform assessment of transcriptome profiling using RNA-seq in the ABRF next-generation sequencing study.
High-throughput RNA sequencing (RNA-seq) greatly expands the potential for genomics discoveries, but the wide variety of platforms, protocols and performance capabilitites has created the need for comprehensive reference data. Here we describe the Association of Biomolecular Resource Facilities next-generation sequencing (ABRF-NGS) study on RNA-seq. We carried out replicate experiments across 15 laboratory sites using reference RNA standards to test four protocols (poly-A-selected, ribo-depleted, size-selected and degraded) on five sequencing platforms (Illumina HiSeq, Life Technologies PGM and Proton, Pacific Biosciences RS and Roche 454). The results show high intraplatform (Spearman rank R > 0.86) and inter-platform (R > 0.83) concordance for expression measures across the deep-count platforms, but highly variable efficiency and cost for splice junction and variant detection between all platforms. For intact RNA, gene expression profiles from rRNA-depletion and poly-A enrichment are similar. In addition, rRNA depletion enables effective analysis of degraded RNA samples. This study provides a broad foundation for cross-platform standardization, evaluation and improvement of RNA-seq.


Extraction of monoclonal antibodies (IgG1) using anionic and anionic/nonionic reverse micelles.
Purification schemes for antibody production based on affinity chromatography are trying to keep pace with increases in cell culture expression levels and many current research initiatives are focused on finding alternatives to chromatography for the purification of Monoclonal antibodies (MAbs). In this article, we have investigated an alternative separation technique based on liquid-liquid extraction called the reverse micellar extraction. We extracted MAb (IgG1) using reverse micelles of an anionic surfactant, sodium bis 2-ethyl-hexyl sulfosuccinate (AOT) and a combination of anionic (AOT) and nonionic surfactants (Brij-30, Tween-85, Span-85) using isooctane as the solvent system. The extraction efficiency of IgG1 was studied by varying parameters, such as pH of the aqueous phase, cation concentration, and type and surfactant concentration. Using the AOT/Isooctane reverse micellar system, we could achieve good overall extraction of IgG1 (between 80 and 90%), but only 30% of the bioactivity of IgG1 could be recovered at the end of the extraction by using its binding to affinity chromatography columns as a surrogate measure of activity. As anionic surfactants were suspected as being one of the reasons for the reduced activity, we decided to combine a nonionic surfactant with an anionic surfactant and then study its effect on the extraction efficiency and bioactivity. The best results were obtained using an AOT/Brij-30/Isooctane reverse micellar system, which gave an overall extraction above 90 and 59% overall activity recovery. An AOT/Tween-85/Isooctane reverse micellar system gave an overall extraction of between 75 and 80% and overall activity recovery of around 40-45%. The results showed that the activity recovery of IgG1 can be significantly enhanced using different surfactant combination systems, and if the recovery of IgG1 can be further enhanced, the technique shows considerable promise for the downstream purification of MAbs.


Invisible fat on CT: making it visible by MRI.
Presence of fat in a lesion significantly narrows the differential diagnosis. Small quantities of macroscopic fat and intracellular fat are invisible on computed tomography (CT) and ultrasonography. Magnetic resonance imaging (MRI) can reveal any fatty change in a lesion and can also differentiate macroscopic fat from intracellular and intravoxel fat. Hypodensity on CT may be a sign of invisible fat and MRI can help to diagnose even minute amounts of fat in liver, pancreas, adrenal, musculoskeletal, and omental pseudolesions and lesions. This article will review the superiority of MRI over CT in demonstrating fat in abdominal lesions.


AIB1 is a predictive factor for tamoxifen response in premenopausal women.
Clinical trials implicate the estrogen receptor (ER) coactivator amplified in breast cancer 1 (AIB1) to be a prognostic and a treatment-predictive factor, although results are not unanimous. We have further investigated this using a controlled randomised trial of tamoxifen versus control. A total of 564 premenopausal women were entered into a randomised study independent of ER status. Using a tissue microarray, AIB1 and ER were analysed by immunohistochemistry. AIB1 scores were obtained from 349 women. High AIB1 correlated to factors of worse prognosis (human epidermal growth factor receptor 2, Nottingham histological grade 3, and lymph node metastases) and to ER negativity. In the control arm, high AIB1 was a negative prognostic factor for recurrence-free survival (RFS) (P = 0.02). However, ER-positive patients with high AIB1 responded significantly to tamoxifen treatment (P = 0.002), increasing RFS to the same level as for systemically untreated patients with low AIB1. Although ER-positive patients with low AIB1 had a better RFS from the beginning, this was not further improved by tamoxifen (P = 0.8). In the control group, high AIB1 was a negative prognostic factor. However, ER-positive patients with high AIB1 responded significantly to tamoxifen. This implicates high AIB1 to be an independent predictive factor of improved response to tamoxifen and not, as has previously been discussed, a factor predicting tamoxifen resistance.


Show and tell: disclosure and data sharing in experimental pathology.
Reproducibility of data from experimental investigations using animal models is increasingly under scrutiny because of the potentially negative impact of poor reproducibility on the translation of basic research. Histopathology is a key tool in biomedical research, in particular for the phenotyping of animal models to provide insights into the pathobiology of diseases. Failure to disclose and share crucial histopathological experimental details compromises the validity of the review process and reliability of the conclusions. We discuss factors that affect the interpretation and validation of histopathology data in publications and the importance of making these data accessible to promote replicability in research.


Steady-state regulation of COPII-dependent secretory cargo sorting by inositol trisphosphate receptors, calcium, and penta EF hand proteins.
Recently, we demonstrated that agonist-stimulated Ca2+ signaling involving IP3 receptors modulates ER export rates through activation of the penta-EF Hand proteins apoptosis-linked gene-2 (ALG-2) and peflin. It is unknown, however, whether IP3Rs and penta-EF proteins regulate ER export rates at steady state. Here we tested this idea in normal rat kidney epithelial cells by manipulation of IP3R isoform expression. Under standard growth conditions, spontaneous cytosolic Ca2+ oscillations occurred simultaneously in successive groups of contiguous cells, generating intercellular Ca2+ waves that moved across the monolayer periodically. Depletion of IP3R-3, typically the least promiscuous IP3R isoform, caused increased cell participation in intercellular Ca2+ waves in unstimulated cells. The increased spontaneous signaling was sufficient to cause increased ALG-2 and COPII coat subunit Sec31A and decreased peflin localization at ER exit sites, resulting in increased ER-to-Golgi transport of the COPII client cargo VSV-G. The elevated ER-to-Golgi transport caused greater concentration of VSV-G at ER exit sites and had reciprocal effects on transport of VSV-G and a bulk-flow cargo, though both cargos equally required Sec31A. Inactivation of client cargo sorting using 4-phenylbutyrate had opposing reciprocal effects on client and bulk-flow cargo and neutralized any effect of ALG-2 activation on transport. This work extends our knowledge of ALG-2 mechanisms and indicates that in normal rat kidney cells, IP3R isoforms regulate homeostatic Ca2+ signaling that helps determine the basal secretion rate and stringency of COPII-dependent cargo sorting.


Paralysis due to the high tackle--a black spot in South African rugby.
The high tackle around the neck is illegal but still commonplace in South African rugby. An analysis of 40 rugby players who sustained spinal cord injury during the period 1985-1989 revealed that 8 were injured by a high tackle. The case histories and radiographs of these 8 players were analysed. The majority sustained flexion-rotation injuries after being tackled from the side. Another mechanism of injury was hyper-extension during a tackle from the rear. Disturbingly, 4 of the 8 players sustained complete permanent paralysis. This was consequent upon the orthopaedic injuries sustained--specifically facet dislocations or 'tear-drop' fractures, both injuries carrying with them a high risk of serious spinal cord injury. It is concluded that foul play in the form of the high tackle is still a major cause of serious spinal cord injury in South African rugby.


Symptom trajectories during chemotherapy in outpatients with lung cancer colorectal cancer, or lymphoma.
Pain, depression, distress, fatigue, and sleep disturbance are common symptoms in oncology patients, but little data are available that examine the trajectories of these symptoms during chemotherapy (CTX). The purposes of this study were to examine the trajectories of these symptoms during the first six cycles of CTX and to determine whether individual characteristics predicted the trajectories of these symptoms. Oncology outpatients (n = 118) with newly diagnosed lung cancer, colorectal cancer, or lymphoma rated symptoms using an electronic patient care monitor system. Pain, fatigue, and sleep disturbance were rated on 0-10 numeric rating scales; depression and distress were evaluated using scales converted to normalized T scores. Latent growth curve analyses (LGCA) examined for intra- and inter-individual differences in the trajectories of these five symptoms during the six cycles of CTX. Symptoms were present at the initiation of CTX (p < 0.0001) for all symptoms (p < 0.05). Distress (p = 0.03) and pain (p = 0.02) intensity decreased significantly over the six cycles of CTX. Advanced disease and a higher number of comorbidities predicted higher fatigue at baseline (p = 0.02 and 0.01 respectively). A diagnosis of lung cancer predicted an increasing intensity of fatigue during CTX (p = 0.04). Concurrent radiation therapy predicted more intense pain over time (p = 0.03). While symptom trajectories were highly variable in patients undergoing initial CTX, the majority of the symptom intensity scores decreased over time. However, patients with lung cancer, those with a higher number of comorbidities, and those with advanced disease experienced more intense fatigue and sleep disturbance over time.


Occurrence of Ochroconis and Verruconis species in clinical specimens from the United States.
Ochroconis is a dematiaceous fungus able to infect immunocompetent people. Recently, the taxonomy of the genus has been reevaluated, and the most relevant species, Ochroconis gallopava, was transferred to the new genus Verruconis. Due to the important clinical implications of these fungi and based on the recent classification, it was of interest to know the spectra of Ochroconis and Verruconis species in clinical samples received in a reference laboratory in the United States. A set of 51 isolates was identified morphologically and molecularly based on sequence analyses of the nuclear ribosomal RNA (nrRNA), actin, and β-tubulin genes. Verruconis gallopava was the most common species (68.6%), followed by Ochroconis mirabilis (21.5%). One isolate of Ochroconis cordanae was found, being reported for the first time in a clinical setting. The most common anatomical site of isolation was the lower respiratory tract (58.8%), followed by superficial and deep tissues at similar frequencies (21.6 and 19.6%, respectively). Interestingly, three new species were found, which are Ochroconis olivacea and Ochroconis ramosa from clinical specimens and Ochroconis icarus of an environmental origin. The in vitro antifungal susceptibilities of eight antifungal drugs against the Ochroconis isolates revealed that terbinafine and micafungin were the most active drugs.


Characterization of speech and language phenotype in children with NRXN1 deletions.
Neurexin 1 gene (NRXN1) deletions are associated with several neurodevelopmental disorders. Communication difficulties have been reported, yet no study has examined specific speech and language features of individuals with NRXN1 deletions. Here, we characterized speech and language phenotypes in 21 children (14 families), aged 1.8-17 years, with NRXN1 deletions. Deletions ranged from 74 to 702 kb and consisted mostly of either exons 1-3 or 1-5. Speech sound disorders were frequent (69%), although few were severe. The majority (57%) of children had difficulty with receptive and/or expressive language, although no homogeneous profiles of deficit were seen across semantic, morphological, or grammatical systems. Social language difficulties were seen in over half the sample (53%). All but two individuals with language difficulties also had intellectual disability/developmental delay. Overall, while speech and language difficulties were common, there was substantial heterogeneity in the severity and type of difficulties observed and no striking communication phenotype was seen. Rather, the speech and language deficits are likely part of broader concomitant neurodevelopmental profiles (e.g., intellectual disability, social skill deficits). Nevertheless, given the high rate of affectedness, it is important speech/language development is assessed so interventions can be applied during childhood in a targeted and timely manner.


Harnessing the Rheological Properties of Liquid Metals To Shape Soft Electronic Conductors for Wearable Applications.
Emerging applications of the Internet of Things in healthcare, wellness, and gaming require continuous monitoring of the body and its environment, fueling the need for wearable devices able to maintain intimate, reliable, and unobtrusive contact with the human body. This translates in the necessity to develop soft and deformable electronics that match the body's mechanics and dynamics. In recent years, various strategies have been proposed to form stretchable circuits and more specifically elastic electrical conductors embedded in elastomeric substrate using either geometrical structuring of solid conductors or intrinsically stretchable materials. Gallium (Ga)-based liquid metals (LMs) are an emerging class of materials offering a particularly interesting set of properties for the design of intrinsically deformable conductors. They concomitantly offer the high electrical conductivity of metals with the ability of liquids to flow and reconfigure. The specific chemical and physical properties of Ga-based LMs differ fundamentally from those of solid conductors and need to be considered to successfully process and implement them into stretchable electronic devices. In this Account, we report on how the key physical and chemical properties of Ga-based LMs can be leveraged to enable repeatable manufacturing and precise patterning of stretchable LM conductors. A comprehensive understanding of the interplay between the LM, its receiving substrate chemistry and topography, and the environmental conditions is necessary to meet the reproducibility and reliability standards for large scale deployment in next-generation wearable systems. In oxidative environments, a solid oxide skin forms at the surface of the LM and provides enough stiffness to counterbalance surface tension, and prevent the LM from beading up to a spherical shape. We review techniques that advantageously harness the oxide skin to form metastable structures such as spraying, 3D printing, or channel injection. Next, we explore how controlling the environmental condition prevents the formation or removes the oxide skin, thereby allowing for selective wetting of Ga lyophilic surfaces. Representative examples include selective plating and physical vapor deposition. The wettability of LMs can be further tuned by engineering the surface chemistry and topology of the receiving substrate to form superlyophobic or superlyophilic surfaces. In particular, our group developed Ga-superlyophilic substrates by engineering the surface of silicone rubber with microstructures and a gold coating layer. Thermal evaporation of Ga on such engineered substrates allows for the formation of smooth LM films with micrometric thickness control and design freedom. The versatility of the available deposition techniques facilitates the implementation of LM conductors in a wide variety of wearable devices. We review various epidermal electronic systems using LM conductors as interconnects to carry power and information, transducers and sensors, antennas, and complex hybrid (soft-rigid) electronic circuits. In addition, we highlight the limitations and challenges inherent to the use of Ga LM conductors that include electromigration, corrosion, solidification, and biocompatibility.


Implications for informatics given expanding access to care for Veterans and other populations.
Recent investigations into appointment scheduling within facilities operated by the US Department of Veterans Affairs (VA) illuminate systemic challenges in meeting its goal of providing timely access to care for all Veterans. In the wake of these investigations, new policies have been enacted to expand access to care at VA facilities as well as non-VA facilities if the VA is unable to provide access within a reasonable timeframe or a Veteran lives more than 40 miles from a VA medical facility. These policies are similar to broader health reform efforts that seek to expand access to care for other vulnerable populations. In this perspective, we discuss the informatics implications of expanded access within the VA and its wider applicability across the US health system. Health systems will require robust health information exchange, to maintain coordination while access to care is expanded. Existing informatics research can guide short-term implementation; furthermore, new research is needed to generate evidence about how best to achieve the long-term aim of expanded access to care.


A platform for the remote conduct of gene-environment interaction studies.
Gene-environment interaction studies offer the prospect of robust causal inference through both gene identification and instrumental variable approaches. As such they are a major and much needed development. However, conducting these studies using traditional methods, which require direct participant contact, is resource intensive. The ability to conduct gene-environment interaction studies remotely would reduce costs and increase capacity. To develop a platform for the remote conduct of gene-environment interaction studies. A random sample of 15,000 men and women aged 50+ years and living in Cardiff, South Wales, of whom 6,012 were estimated to have internet connectivity, were mailed inviting them to visit a web-site to join a study of successful ageing. Online consent was obtained for questionnaire completion, cognitive testing, re-contact, record linkage and genotyping. Cognitive testing was conducted using the Cardiff Cognitive Battery. Bio-sampling was randomised to blood spot, buccal cell or no request. A heterogeneous sample of 663 (4.5% of mailed sample and 11% of internet connected sample) men and women (47% female) aged 50-87 years (median=61 yrs) from diverse backgrounds (representing the full range of deprivation scores) was recruited. Bio-samples were donated by 70% of those agreeing to do so. Self report questionnaires and cognitive tests showed comparable distributions to those collected using face-to-face methods. Record linkage was achieved for 99.9% of participants. This study has demonstrated that remote methods are suitable for the conduct of gene-environment interaction studies. Up-scaling these methods provides the opportunity to increase capacity for large-scale gene-environment interaction studies.


Mouse model of hemolytic-uremic syndrome caused by endotoxin-free Shiga toxin 2 (Stx2) and protection from lethal outcome by anti-Stx2 antibody.
Hemolytic-uremic syndrome (HUS) results from infection by Shiga toxin (Stx)-producing Escherichia coli and is the most common cause of acute renal failure in children. We have developed a mouse model of HUS by administering endotoxin-free Stx2 in multiple doses over 7 to 8 days. At sacrifice, moribund animals demonstrated signs of HUS: increased blood urea nitrogen and serum creatinine levels, proteinuria, deposition of fibrin(ogen), glomerular endothelial damage, hemolysis, leukocytopenia, and neutrophilia. Increased expression of proinflammatory chemokines and cytokines in the sera of Stx2-treated mice indicated a systemic inflammatory response. Currently, specific therapeutics for HUS are lacking, and therapy for patients is primarily supportive. Mice that received 11E10, a monoclonal anti-Stx2 antibody, 4 days after starting injections of Stx2 recovered fully, displaying normal renal function and normal levels of neutrophils and lymphocytes. In addition, these mice showed decreased fibrin(ogen) deposition and expression of proinflammatory mediators compared to those of Stx2-treated mice in the absence of antibody. These results indicate that, when performed during progression of HUS, passive immunization of mice with anti-Stx2 antibody prevented the lethal effects of Stx2.


Acupuncture modulates resting state connectivity in default and sensorimotor brain networks.
Previous studies have defined low-frequency, spatially consistent networks in resting fMRI data which may reflect functional connectivity. We sought to explore how a complex somatosensory stimulation, acupuncture, influences intrinsic connectivity in two of these networks: the default mode network (DMN) and sensorimotor network (SMN). We analyzed resting fMRI data taken before and after verum and sham acupuncture. Electrocardiography data were used to infer autonomic modulation through measures of heart rate variability (HRV). Probabilistic independent component analysis was used to separate resting fMRI data into DMN and SMN components. Following verum, but not sham, acupuncture there was increased DMN connectivity with pain (anterior cingulate cortex (ACC), periaqueductal gray), affective (amygdala, ACC), and memory (hippocampal formation, middle temporal gyrus) related brain regions. Furthermore, increased DMN connectivity with the hippocampal formation, a region known to support memory and interconnected with autonomic brain regions, was negatively correlated with acupuncture-induced increase in a sympathetic related HRV metric (LFu), and positively correlated with a parasympathetic related metric (HFu). Following verum, but not sham, acupuncture there was also increased SMN connectivity with pain-related brain regions (ACC, cerebellum). We attribute differences between verum and sham acupuncture to more varied and stronger sensations evoked by verum acupuncture. Our results demonstrate for the first time that acupuncture can enhance the post-stimulation spatial extent of resting brain networks to include anti-nociceptive, memory, and affective brain regions. This modulation and sympathovagal response may relate to acupuncture analgesia and other potential therapeutic effects.


Reducing coronary risk by raising HDL-cholesterol: risk modelling the addition of nicotinic acid to existing therapy.
Reduction in total cholesterol (TC) and LDL-cholesterol (LDL-C) forms one of the principal objectives of most cardiovascular secondary prevention strategies. Many patients being treated with statins, however, have significant residual dyslipidaemia, with many having suboptimal HDL-cholesterol (HDL-C) levels. The addition of nicotinic acid to a statin has been shown to improve this profile, although clinical outcome evidence is currently lacking. This study set out to model the impact of nicotinic acid therapy on cardiovascular risk in these patients, based on Framingham risk assessments on a cohort of patients drawn from UK general practitioner records. Cardiovascular risk profiles were extracted from a research database of 602,222 patients from 98 UK general practices. 23 262 statin-treated patients with established cardiovascular disease or diabetes were identified and their 4-year Framingham risk was estimated. Patients who had either TC or HDL-C outside the desirable range then had their lipid profile adjusted in accordance with the likely performance of nicotinic acid, and the Framingham risk was then re-assessed. Baseline 4-year coronary risk in the group as a whole was 11.5% (95%CI: 11.4-11.6). After adjustment of the lipid profile, this was reduced to 9.7% (95%CI: 9.6-9.8), a reduction in risk of 15.9% (95%CI: 15.1-16.6). When modelling was limited to those with diabetes or an abnormal treated lipid profile, the magnitude of change was increased to 23-29% depending on sex and subgroup. Risk factor modelling suggests that raising HDL-C levels using nicotinic acid in statin-treated patients is likely to yield significant incremental clinical benefits. The results of clinical trials currently under way are awaited with interest.


The Swiss National Vaccination Coverage Survey, 2005-2007.
We described the results from the Swiss National Vaccination Coverage Survey (SNVCS) 2005-2007, a survey designed to monitor immunization coverage of children and adolescents residing in Switzerland in each canton within a three-year period. The SNVCS is a cross-sectional survey using a two-stage sampling design targeting children aged 2, 8, and 16 years. Families of selected children were contacted by mail and telephone. Coverage was determined via vaccination cards or vaccination summary forms. A total of 25 out of 26 cantons participated in the survey, with 8,286 respondents for children aged 24-35 months, 10,314 respondents for children aged 8 years, and 9,301 respondents for teenagers aged 16 years. Compared with data from 1999-2003, coverage estimates for toddlers remained unchanged for diphtheria, tetanus, pertussis, poliomyelitis, and Haemophilus influenzae type b vaccines at three doses, but increased five percentage points to 86%-87% for measles-mumps-rubella at one dose and was 71% at two doses. Coverage for measles, mumps, and rubella were 89%-90% at one dose and 75% at two doses for 8-year-olds, and 94% and 76% for the two dosages, respectively, for 16-year-olds. Linguistic region and nationality were highly correlated with being vaccinated against measles for the two younger age groups. Despite the increase in vaccine coverage, measles vaccination is still low, and the World Health Organization goal to eliminate measles by 2010 was not achieved in Switzerland. More efforts are needed by the cantons and the central government to increase vaccination coverage.


Elastogenesis in human arterial disease: a role for macrophages in disordered elastin synthesis.
Elastin, an extracellular matrix protein, constitutes about 30% of the dry weight of the arteries. Elastolysis induced by inflammatory processes is active in chronic arterial diseases. However, elastogenesis in arterial diseases has received little attention. In this work we hypothesized that disordered elastogenesis is active in matrix remodeling in atheroma and abdominal aortic aneurysm (AAA). Human AAA and atheroma have 4- to 6-fold more tropoelastin protein than nondiseased arteries. The smooth muscle cell-containing media and fibrous cap of atherosclerotic arteries contain ordered mature elastin, whereas macrophage (MPhi)-rich regions often have disorganized elastic fibers. Surprisingly, in addition to smooth muscle cells, MPhis in diseased arteries also produce the elastin precursor tropoelastin, as shown by double immunostaining, in situ hybridization, and reverse transcription-polymerase chain reaction for tropoelastin mRNA. Cultured monocyte-derived MPhis can express the elastin gene. AAA have 9-fold but atheroma only 1.6-fold lower levels of desmosine, a marker for mature cross-linked elastin, than normal arteries. This study demonstrates ongoing but often ineffective elastogenesis in arterial disease and establishes human macrophages as a novel source for this important matrix protein. These results have considerable import for understanding mechanisms of extracellular matrix remodeling in arterial diseases.


Dendritic cell-based immunotherapy targeting Wilms' tumor 1 in patients with recurrent malignant glioma.
Dendritic cell (DC)-based vaccination is considered a potentially effective therapy against advanced cancer. The authors conducted a Phase I study to investigate the safety and immunomonitoring of Wilms' tumor 1 (WT1)-pulsed DC vaccination therapy for patients with relapsed malignant glioma. WT1-pulsed and/or autologous tumor lysate-pulsed DC vaccination therapy was performed in patients with relapsed malignant gliomas. Approximately 1 × 10(7) to 2 × 10(7) pulsed DCs loaded with WT1 peptide antigen and/or tumor lysate were intradermally injected into the axillary areas with OK-432, a streptococcal preparation, at 2-week intervals for at least 5-7 sessions (1 course) during an individual chemotherapy regimen. Ten patients (3 men, 7 women; age range 24-64 years [median 39 years]) with the following tumors were enrolled: glioblastoma (6), anaplastic astrocytoma (2), anaplastic oligoastrocytoma (1), and anaplastic oligodendroglioma (1). Modified WT1 peptide-pulsed DC vaccine was administered to 7 patients, tumor lysate-pulsed DC vaccine to 2 patients, and both tumor lysate-pulsed and WT1-pulsed DC vaccine to 1 patient. The clinical response was stable disease in 5 patients with WT1-pulsed DC vaccination. In 2 of 5 patients with stable disease, neurological findings improved, and MR images showed tumor shrinkage. No serious adverse events occurred except Grade 1-2 erythema at the injection sites. WT1 tetramer analysis detected WT1-reactive cytotoxic T cells after vaccination in patients treated with WT1-pulsed therapy. Positivity for skin reaction at the injection sites was 80% (8 of 10 patients) after the first session, and positivity remained for these 8 patients after the final session. This study of WT1-pulsed DC vaccination therapy demonstrated safety, immunogenicity, and feasibility in the management of relapsed malignant gliomas.


Longitudinal Changes in Component Processes of Working Memory.
Working memory (WM) entails maintenance and manipulation of information in the absence of sensory input. This study investigated the trajectories and neural basis of these component processes of WM functions in aging. Longitudinal human functional magnetic resonance imaging (fMRI) data are presented from 136 older individuals (55-80 years) who were scanned at baseline and again 4 years later. We obtained evidence that age-related changes in parietal and frontal components of the WM core network are dissociable in terms of their role in maintenance of perceptual representations and further manipulation of this information, respectively. Individual difference analyses in performance subgroups showed that only prefrontal changes in fMRI activation were accompanied by changes in performance, but parietal brain activity was related to study dropout. We discuss the results in terms of possible neurobiological causes underlying separable aging-related declines in inferior parietal cortex and lateral prefrontal cortex that differentially affect WM functions.


Efficacy of lower limb compression and combined treatment of manual massage and lower limb compression on symptoms of exercise-induced muscle damage in women.
Strategies to manage the symptoms of exercise-induced muscle damage (EIMD) are widespread, though are often based on anecdotal evidence. The aim of this study was to determine the efficacy of a combination of manual massage and compressive clothing and compressive clothing individually as recovery strategies after muscle damage. Thirty-two female volunteers completed 100 plyometric drop jumps and were randomly assigned to a passive recovery (n = 17), combined treatment (n = 7), or compression treatment group (n = 8). Indices of muscle damage (perceived soreness, creatine kinase activity, isokinetic muscle strength, squat jump, and countermovement jump performance) were assessed immediately before and after 1, 24, 48, 72, and 96 hours of plyometric exercise. The compression treatment group wore compressive tights for 12 hours after damage and the combined treatment group received a 30-minute massage immediately after damaging exercise and wore compression stockings for the following 11.5 hours. Plyometric exercise had a significant effect on all indices of muscle damage (p < 0.05). The treatments significantly reduced decrements in isokinetic muscle strength, squat jump performance, and countermovement jump performance and reduced the level of perceived soreness in comparison with the passive recovery group (p < 0.05). The addition of sports massage to compression after muscle damage did not improve performance recovery, with recovery trends being similar in both treatment groups. The treatment combination of massage and compression significantly moderated perceived soreness at 48 and 72 hours after plyometric exercise (p < 0.05) in comparison with the passive recovery or compression alone treatment. The results indicate that the use of lower limb compression and a combined treatment of manual massage with lower limb compression are effective recovery strategies following EIMD. Minimal performance differences between treatments were observed, although the combination treatment may be beneficial in controlling perceived soreness.


Rat pancreatic kallikrein mRNA: nucleotide sequence and amino acid sequence of the encoded preproenzyme.
We have cloned via recombinant DNA technology the mRNA sequence for rat pancreatic preprokallikrein. Four cloned overlapping double-stranded cDNAs gave a continuous mRNA sequence of 867 nucleotides beginning within the 5'-noncoding region and extending to the poly(A) tail. The mRNA sequence reveals that pancreatic kallikrein is synthesized as a prezymogen of 265 amino acids, including a proposed secretory prepeptide of 17 amino acids and a proposed activation peptide of 11 amino acids. The activation peptide, although similar in length, is distinct from those of the other classes of pancreatic serine proteases. The amino acid sequence of the predicted active form of the enzyme is closely related to the partial sequences obtained for other kallikrein-like serine proteases including rat submaxillary gland kallikrein, pig pancreatic and submaxillary gland kallikreins, the gamma subunit of mouse nerve growth factor, and rat tonin. Key amino acid residues thought to be involved in the substrate-cleavage specificity of kallikreins are retained. Hybridization analysis showed relatively high levels of kallikrein mRNA in the rat pancreas, submaxillary and parotid glands, spleen, and kidney, indicating the active synthesis of kallikrein in these tissues.


Exact direct-space asymmetric units for the 230 crystallographic space groups.
It is well known that the direct-space asymmetric unit definitions found in the International Tables for Crystallography, Volume A, are inexact at the borders. Face- and edge-specific sub-conditions have to be added to remove parts redundant under symmetry. This paper introduces a concise geometric notation for asymmetric unit conditions. The notation is the foundation for a reference table of exact direct-space asymmetric unit definitions for the 230 crystallographic space-group types. The change-of-basis transformation law for the conditions is derived, which allows the information from the reference table to be used for any space-group setting. We also show how the vertices of an asymmetric unit can easily be computed from the information in the reference table.


Protein C deficiency resulting from possible double heterozygosity and its response to danazol.
A unique family with protein C (PC) deficiency is described. The proband had a history of renal vein thrombosis as a newborn and iliofemoral thrombosis at the age of 6 years. After 6 months of heparin treatment, discontinuation of anticoagulation therapy was accompanied by persistent hypofibrinogenemia with increased fibrinogen consumption. With continuous infusion of heparin, fibrinogen turnover normalized, and the child has remained free of thrombosis. Both the immunologic level of PC and the functional activity measured by amidolytic assay were moderately reduced (47% and 34%, respectively). Functional activity of PC measured by its anticoagulant activity was disproportionately lower (14%). A 3-year-old asymptomatic sibling had a similar disproportionate reduction of PC anticoagulant activity compared with the amidolytic activity or immunologic level. The mother demonstrated type I PC deficiency with a proportionate reduction in immunologic protein levels (59%), anticoagulant activity (52%), and amidolytic activity (46%), whereas the father had type II PC deficiency with normal immunologic protein levels (102%), normal amidolytic function (98%), but a low anticoagulant function (50%). An abnormal PC molecule was detected by two-dimensional immunoelectrophoresis in the father and two children. These data are consistent with the hypothesis that the children are doubly heterozygous for two different types of PC deficiency inherited from each of the parents. A 14-day trial of danazol in the proband resulted in a rise in the PC antigen concentration from 66% to 98% but no change in PC anticoagulant function.