Nitrates for acute heart failure syndromes.
Current drug therapy for acute heart failure syndromes (AHFS) consists mainly of diuretics supplemented by vasodilators or inotropes. Nitrates have been used as vasodilators in AHFS for many years and have been shown to improve some aspects of AHFS in some small studies. The aim of this review was to determine the clinical efficacy and safety of nitrate vasodilators in AHFS. To quantify the effect of different nitrate preparations (isosorbide dinitrate and nitroglycerin) and the effect of route of administration of nitrates on clinical outcome, and to evaluate the safety and tolerability of nitrates in the management of AHFS. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2011, Issue 3), MEDLINE (1950 to July week 2 2011) and EMBASE (1980 to week 28 2011). We searched the Current Controlled Trials MetaRegister of Clinical Trials (compiled by Current Science) (July 2011). We checked the reference lists of trials and contacted trial authors. We imposed no language restriction. Randomised controlled trials comparing nitrates (isosorbide dinitrate and nitroglycerin) with alternative interventions (frusemide and morphine, frusemide alone, hydralazine, prenalterol, intravenous nesiritide and placebo) in the management of AHFS in adults aged 18 and over. Two authors independently performed data extraction. Two authors performed trial quality assessment. We used mean difference (MD), odds ratio (OR) and 95% confidence intervals (CI) to measure effect sizes. Two authors independently assessed and rated the methodological quality of each trial using the Cochrane Collaboration tool for assessing risk of bias. Four studies (634 participants) met the inclusion criteria. Two of the included studies included only patients with AHFS following acute myocardial infarction (AMI); one study excluded patients with overt AMI; and one study included participants with AHFS with and without acute coronary syndromes.Based on a single study, there was no significant difference in the rapidity of symptom relief between intravenous nitroglycerin/N-acetylcysteine and intravenous frusemide/morphine after 30 minutes (fixed-effect MD -0.30, 95% CI -0.65 to 0.05), 60 minutes (fixed-effect MD -0.20, 95% CI -0.65 to 0.25), three hours (fixed-effect MD 0.20, 95% CI -0.27 to 0.67) and 24 hours (fixed-effect MD 0.00, 95% CI -0.31 to 0.31). There is no evidence to support a difference in AHFS patients receiving intravenous nitrate vasodilator therapy or alternative interventions with regard to the following outcome measures: requirement for mechanical ventilation, systolic blood pressure (SBP) change after three hours and 24 hours, diastolic blood pressure (DBP) change after 30, 60 and 90 minutes, heart rate change at 30 minutes, 60 minutes, three hours and 24 hours, pulmonary artery occlusion pressure (PAOP) change after three hours and 18 hours, cardiac output (CO) change at 90 minutes and three hours and progression to myocardial infarction. There is a significantly higher incidence of adverse events after three hours with nitroglycerin compared with placebo (odds ratio 2.29, 95% CI 1.26 to 4.16) based on a single study. There was no consistent evidence to support a difference in AHFS patients receiving intravenous nitrate vasodilator therapy or alternative interventions with regard to the following secondary outcome measures: SBP change after 30 and 60 minutes, heart rate change after 90 minutes, and PAOP change after 90 minutes. None of the included studies reported healthcare costs as an outcome measure. There were no data reported by any of the studies relating to the acceptability of the treatment to the patients (patient satisfaction scores).Overall there was a paucity of relevant quality data in the included studies. Assessment of overall risk of bias in these studies was limited as three of the studies did not give sufficient detail to allow assessment of potential risk of bias. There appears to be no significant difference between nitrate vasodilator therapy and alternative interventions in the treatment of AHFS, with regard to symptom relief and haemodynamic variables. Nitrates may be associated with a lower incidence of adverse effects after three hours compared with placebo. However, there is a lack of data to draw any firm conclusions concerning the use of nitrates in AHFS because current evidence is based on few low-quality studies.


Summary health statistics for U.S. adults: National Health Interview Survey, 2008.
This report presents health statistics from the 2008 National Health Interview Survey (NHIS) for the civilian noninstitutionalized adult population, classified by sex, age, race and ethnicity, education, family income, poverty status, health insurance coverage, marital status, and place and region of residence. Estimates are presented for selected chronic conditions and mental health characteristics, functional limitations, health status, health behaviors, health care access and utilization, and human immunodeficiency virus testing. Percentages and percent distributions are presented in both age-adjusted and unadjusted versions. NHIS is a household, multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention's National Center for Health Statistics. In 2008, data were collected on 21,781 adults in the Sample Adult questionnaire. The conditional response rate was 74.2%, and the final response rate was 62.6%. The health information for adults in this report was obtained from one randomly selected adult per family. In very rare instances where the sample adult was not able to respond for himself or herself, a proxy was used. In 2008, 61% of adults 18 years of age or over reported excellent or very good health. Fifty-nine percent of adults had never participated in any type of vigorous leisure-time physical activity, and 16% of adults did not have a usual place of health care. Twelve percent of adults had been told by a doctor or health professional that they had heart disease, and 24% had been told on two or more visits that they had hypertension. Twenty-one percent of all adults were current smokers, and 21% were former smokers. Based on estimates of body mass index, 35% of adults were overweight and 27% were obese.


Bees, honey and pollen as sentinels for lead environmental contamination.
Three beehive matrices, sampled in eighteen apiaries from West France, were analysed for the presence of lead (Pb). Samples were collected during four different periods in both 2008 and 2009. Honey was the matrix the least contaminated by Pb (min = 0.004 μg g(-1); max = 0.378 μg g(-1); mean = 0.047 μg g(-1); sd = 0.057). The contamination of bees (min = 0.001 μg g(-1); max = 1.869 μg g(-1); mean = 0.223 μg g(-1); sd = 0.217) and pollen (min = 0.004 μg g(-1); max = 0.798 μg g(-1); mean = 0.240 μg g(-1); sd = 0.200) showed similar levels and temporal variations but bees seemed to be more sensitive bringing out the peaks of Pb contamination. Apiaries in urban and hedgerow landscapes appeared more contaminated than apiaries in cultivated and island landscapes. Sampling period had a significant effect on Pb contamination with higher Pb concentrations determined in dry seasons.


Evaluation of the presence of selected viral and bacterial nucleic acids in pericardial samples from dogs with or without idiopathic pericardial effusion.
Many viruses have been identified in pericardial fluid and in tissue samples from humans with pericarditis by means of molecular diagnostics. In canine idiopathic pericardial effusion there is as yet no conclusive evidence to support the involvement of an infectious agent. This study was designed to investigate a possible relationship between idiopathic pericardial effusion in dogs and viruses most commonly encountered in humans affected with viral pericarditis. Coxsackievirus B3 RNA, influenza virus type A RNA, human adenovirus type 2 DNA, human cytomegalovirus DNA, and parvovirus B19 DNA were investigated using PCR on pericardial effusion samples and pericardial tissue specimens collected from 14 dogs with idiopathic pericardial effusion. PCR was also used to test for two bacteria, Borrelia burgdorferi and Chlamydia pneumoniae. The same microorganisms were also looked for in pericardial effusions or pericardial washes from 10 dogs with neoplastic pericardial effusion, and in samples collected from 10 dogs which died of a non-cardiac disease. One pericardial effusion sample from a dog with the idiopathic form of the disease tested positive for influenza virus type A and sequencing of the amplicon confirmed the PCR result. In another dog from the same group a cytomegalovirus was detected by PCR in the effusion, but sequencing showed this to be a false-positive result. The genomes of the microorganisms investigated were not detected in neoplastic effusions or pericardial washes. The results indicate that viral and bacterial DNA/RNA of relevance for human pericarditis is rare in pericardial samples from dogs with idiopathic pericardial effusion. The finding of influenza type A viral RNA in pericardial fluid from one dog with the idiopathic form of the disease warrants further investigation.


Leasing a medical curriculum: What's it worth?
Introduction: The early part of this century saw an unprecedented growth in number and size of Australian medical schools. There was some partnering of the new schools with existing programs. Griffith, Deakin and Curtin Universities leased an established curriculum from Flinders University. Nature and rationale for curriculum leasing: The new schools had short startup times and leasing a curriculum enabled them to appoint key staff, develop facilities and meet accreditation requirements in a timely way. However, the lease arrangements were costly and the curriculum was largely determined before the Dean and key staff appointments. Outcomes of leasing: There was differential adoption of the leased curriculum. The first two years of the courses at Flinders were transferred with little change. The final two years of predominantly clinical studies were developed differently. This is explained through Michael Fullan's work on context in educational change. The context of the clinical years of the courses involved negotiations with local health services and other schools using those health services. The advantage of the leasing arrangements was that the new schools could proceed through early development and accreditation, while having time and opportunity to negotiate a clinical curriculum that engaged local health services and fulfilled the new schools' missions.


"Messing with the mind": evolutionary challenges to human brain augmentation.
The issue of brain augmentation has received considerable scientific attention over the last two decades. A key factor to brain augmentation that has been widely overlooked are the complex evolutionary processes which have taken place in evolving the human brain to its current state of functioning. Like other bodily organs, the human brain has been subject to the forces of biological adaptation. The structure and function of the brain, is very complex and only now we are beginning to understand some of the basic concepts of cognition. Therefore, this article proposes that brain-machine interfacing and nootropics are not going to produce "augmented" brains because we do not understand enough about how evolutionary pressures have informed the neural networks which support human cognitive faculties.


Creation of CRISPR-based germline-genome-engineered mice without ex vivo handling of zygotes by i-GONAD.
Methods to create genetically engineered mice involve three major steps: harvesting embryos from one set of females, microinjection of reagents into embryos ex vivo and their surgical transfer to another set of females. Although tedious, these methods have been used for more than three decades to create mouse models. We recently developed a method named GONAD (genome editing via oviductal nucleic acids delivery), which bypasses these steps. GONAD involves injection of CRISPR components (Cas9 mRNA and guide RNA (gRNA)) into the oviducts of pregnant females 1.5 d post conception, followed by in vivo electroporation to deliver the components into the zygotes in situ. Using GONAD, we demonstrated that target genes can be disrupted and analyzed at different stages of mouse embryonic development. Subsequently, we developed improved GONAD (i-GONAD) by delivering CRISPR ribonucleoproteins (RNPs; Cas9 protein or Cpf1 protein and gRNA) into day-0.7 pregnant mice, which made it suitable for routine generation of knockout and large-deletion mouse models. i-GONAD can also generate knock-in models containing up to 1-kb inserts when single-stranded DNA (ssDNA) repair templates are supplied. i-GONAD offers other advantages: it does not require vasectomized males and pseudo-pregnant females, the females used for i-GONAD are not sacrificed and can be used for other experiments, it can be easily adopted in laboratories lacking sophisticated microinjection equipment, and can be implemented by researchers skilled in small-animal surgery but lacking embryo-handling skills. Here, we provide a step-by-step protocol for establishing the i-GONAD method. The protocol takes ∼6 weeks to generate the founder mice.


Experimental comparison of two methods to study barley responses to partial submergence.
Crop yield is dependent on climate conditions, which are becoming both more variable and extreme in some areas of the world as a consequence of global climate change. Increased precipitation and flooding events are the cause of important yield losses due to waterlogging or (partial) submergence of crops in the field. Our ability to screen efficiently and quickly for varieties that have increased tolerance to waterlogging or (partial) submergence is important. Barley, a staple crop worldwide, is particularly sensitive to waterlogging. Screening for waterlogging tolerant barley varieties has been ongoing for many years, but methods used to screen vary greatly, from the type of soil used to the time at which the treatment is applied. This variation makes it difficult to cross-compare results. Here, we have devised a scoring system to assess barley tolerance to waterlogging and compare two different methods when partial submergence is applied with either water or a starch solution at an early developmental stage, which is particularly sensitive to waterlogging or partial submergence. The use of a starch solution has been previously shown to result in more reducing soil conditions and has been used to screen for waterlogging tolerance. Our results show that the two methods provide similar results to qualitatively rank varieties as tolerant or sensitive, while also affecting plants differently, in that application of a starch solution results in stronger and earlier symptoms than applying partial submergence with water.


Habitual cocoa intake reduces arterial stiffness in postmenopausal women regardless of intake frequency: a randomized parallel-group study.
Arterial stiffness is substantially higher in postmenopausal than in premenopausal women. Daily cocoa intake has been shown to reduce central arterial stiffness in health adults, regardless of age; however, the effect of cocoa-intake frequency on arterial stiffness in postmenopausal women remains unclear. Therefore, the purpose of this study was to investigate the effects of cocoa-intake frequency on arterial stiffness in postmenopausal women. A total of 26 postmenopausal women (mean age ± standard deviation 64±12 years) were randomly assigned to two groups with different cocoa-intake frequencies: one group ingested 17 g of cocoa once daily except on Sundays (every-day group, n=13), and the other ingested 17 g of cocoa twice daily every other day (every-other-day group, n=13). These intake regimens were maintained in both groups for 12 weeks. Carotid-femoral pulse-wave velocity and femoral-ankle pulse-wave velocity were measured in both groups at baseline and again at the end of the 12-week study period. Compared to baseline, both pulse-wave velocities had significantly decreased after the 12-week study period in both groups (P<0.05). However, no significant difference in degree of change was observed between the two groups. Although this study did not include a sedentary control group, these results suggest that regardless of frequency, habitual cocoa intake reduces central and peripheral arterial stiffness in postmenopausal women.


Precise diagnosis in different scenarios using photoacoustic and fluorescence imaging with dual-modality nanoparticles.
Photoacoustic imaging and fluorescence molecular imaging are emerging as important research tools for biomedical studies. Photoacoustic imaging offers both strong optical absorption contrast and high ultrasonic resolution, and fluorescence molecular imaging provides excellent superficial resolution, high sensitivity, high throughput, and the ability for real-time imaging. Therefore, combining the imaging information of both modalities can provide comprehensive in vivo physiological and pathological information. However, currently there are limited probes available that can realize both fluorescence and photoacoustic imaging, and advanced biomedical applications for applying this dual-modality imaging approach remain underexplored. In this study, we developed a dual-modality photoacoustic-fluorescence imaging nanoprobe, ICG-loaded Au@SiO2, which was uniquely designed, consisting of gold nanorod cores and indocyanine green with silica shell spacer layers to overcome fluorophore quenching. This nanoprobe was examined by both PAI and FMI for in vivo imaging on tumor and ischemia mouse models. Our results demonstrated that the nanoparticles can specifically accumulate at the tumor and ischemic areas and be detected by both imaging modalities. Moreover, this dual-modality imaging strategy exhibited superior advantages for a precise diagnosis in different scenarios. The new nanoprobe with the dual-modality imaging approach holds great potential for diagnosis and stage classification of tumor and ischemia related diseases.


Aqueous solution behaviour and solubilisation properties of octadecyl cationic gemini surfactants and their comparison with their amide gemini analogues.
Gemini surfactants 18-s-18(Et), comprised of two ethylammonium headgroups and two alkyl tails with m = 18 carbon atoms with spacers of s = 4, 6, 8 and 10 linking the headgroups (alkanediyl-α,ω-bis(diethyloctadecylammonium bromides)), were obtained. Their aqueous solution behaviour, including adsorption at the interface and aggregation in solution, was followed by tensiometric, conductometric and spectroscopic methods. The critical micelle concentration (CMC) of the surfactants decreased with increasing spacer length. The size of 18-s-18(Et) aggregates formed at concentrations of 10 and 40 CMC measured by DLS varied with the elongation of the spacer. Visualisation of aggregated surfactant structures at 40 CMC by cryo-TEM evidenced the formation of different morphologies depending on spacer length. Gemini with s = 4 formed elongated, cylindrical micelles, while geminis of s = 6, 8 and 10 self-assembled into vesicles. The ability of the studied geminis to solubilise hydrophobic dye Sudan I in water was determined as a function of surfactant concentration, demonstrating their high efficiency. Results for 18-s-18(Et) geminis were compared with those previously obtained for their analogues containing an amide group placed between headgroups and tails. The significant impact of amide groups on the surface activity and aggregation properties of gemini surfactants was evidenced and is related to hydrogen-bond formation by amide-containing compounds.


Immunosuppressive therapy for severe chronic uveitis.
Twenty-five patients with severe, progressive, chronic uveitis who were poorly responsive or unresponsive to corticosteroid therapy received low-dose prednisone and cytotoxic immunosuppressive therapy. Azathioprine (2.0 to 2.5 mg/kg) or chlorambucil (6 to 8 mg) was combined with prednisone (10 to 15 mg) daily in a long-term therapeutic program. All 25 patients exhibited a therapeutic response. In 18 of 25 patients, complete quiescence of the inflammatory process was observed. The remaining seven patients showed a substantial decrease of uveitis with persistence of some inflammatory changes. Adverse side effects that resulted from this therapy were infrequent.


Alterations in IL-4, IL-10 and IFN-γ levels synergistically decrease lipid content and protein expression of FAS and mature SREBP-1 in human sebocytes.
When anti-acne alternatives from dietary and plant sources are ingested, systemic alterations of interleukin (IL)-4, IL-10, IL-12 and interferon (IFN)-γ, individually or simultaneously, are induced at a 0.1-10.0-fold (×) range of normal physiological concentrations (1×). However, little is known about the effects of these cytokines on excess sebum, a pathophysiological factor of acne development. In this study, human sebocytes were treated with 0.1-10.0× of IL-4, IL-10, IL-12 and IFN-γ for 3 or 5 days to elucidate the effects on lipid content. Treatment with individual cytokines decreased the lipid content at specific concentrations rather than in a concentration-dependent manner. Specifically, 5.0× of IL-4, 5.0× of IFN-γ (5.0IFN), and 0.5×, 5.0× and 10.0× of IL-10 for 3 days, and 0.5× of IL-4 (0.5IL4) for 5 days decreased lipid content to 87.6-93.0% of the control. Treatment with other concentrations of IL-4, IL-10 and IFN-γ, and 0.1-10.0× of IL-12 did not alter lipid content. Combined treatment with 0.5IL4, 5.0IFN and 0.5× of IL-10 for 3 or 5 days decreased the lipid content more than each individual treatment. However, this effect was more evident after 3 days, in parallel with decreased levels of triglycerides, cholesterol esters and free fatty acids, the major lipid compositions of sebocytes, and decreased protein expression of fatty acid synthase (FAS) and mature sterol response element-binding protein-1 (SREBP-1), the lipogenesis-related factors, without altered cell proliferation. We demonstrated that suppressed IL-4 and IL-10 with enhanced IFN-γ synergistically decreased lipid content and protein expression of FAS and mature SREBP-1 in human sebocytes.


Hepsin, a cell surface serine protease identified in hepatoma cells, is overexpressed in ovarian cancer.
Extracellular proteases mediate the digestion of neighboring extracellular matrix components in initial tumor growth, allow shedding or desquamation of tumor cells into the surrounding environment, provide the basis for invasion of basement membranes in target metastatic organs, and are required for release and activation of many growth and angiogenic factors. We identified overexpression of the serine protease hepsin gene in ovarian carcinomas and investigated the expression of this gene in 44 ovarian tumors (12 low malignant potential tumors and 32 carcinomas) and 10 normal ovaries. Quantitative PCR was used to determine the relative expression of hepsin compared to that of beta-tubulin. The mRNA expression levels of hepsin were significantly elevated in 7 of 12 low malignant potential tumors and in 27 of 32 carcinomas. On Northern blot analysis, the hepsin transcript was abundant in carcinoma but was almost never expressed in normal adult tissue, including normal ovary. Our results suggest that hepsin is frequently overexpressed in ovarian tumors and therefore may be a candidate protease in the invasive process and growth capacity of ovarian tumor cells.


Bone mineral density changes in relation to environmental PCB exposure.
Bone toxicity has been linked to organochlorine exposure following a few notable poisoning incidents, but epidemiologic studies in populations with environmental organochlorine exposure have yielded inconsistent results. The aim of this study was to investigate whether organochlorine exposure was associated with bone mineral density (BMD) in a population 60-81 years of age (154 males, 167 females) living near the Baltic coast, close to a river contaminated by polychlorinated biphenyls (PCBs). We measured forearm BMD in participants using dual energy X-ray absorptiometry; and we assessed low BMD using age- and sex-standardized Z-scores. We analyzed blood samples for five dioxin-like PCBs, the three most abundant non-dioxin-like PCBs, and p,p'-dichloro-phenyldichloroethylene (p,p'-DDE). In males, dioxin-like chlorobiphenyl (CB)-118 was negatively associated with BMD; the odds ratio for low BMD (Z-score less than -1) was 1.06 (95% confidence interval, 1.01-1.12) per 10 pg/mL CB-118. The sum of the three most abundant non-dioxin-like PCBs was positively associated with BMD, but not with a decreased risk of low BMD. In females, CB-118 was positively associated with BMD, but this congener did not influence the risk of low BMD in women. Environmental organochlorine exposures experienced by this population sample since the 1930s in Sweden may have been sufficient to result in sex-specific changes in BMD.


Regional choroidal blood flow and multifocal electroretinography in experimental glaucoma in rhesus macaques.
To test a hypothesis of regional variation in the effect of experimental glaucoma on choroidal blood flow (ChBF) and retinal function. Five rhesus macaques underwent laser trabecular destruction (LTD) to induce elevated intraocular pressure (IOP). Intraocular pressures were elevated for 56 to 57 weeks. Multifocal electroretinographic (mfERG) and multifocal visual evoked cortical potential (mfVEP) testing were performed at regular intervals before and during the period of IOP elevation. At euthanasia, the IOP was manometrically controlled at 35 (experimentally glaucomatous eye) and 15 (fellow control eye) mm Hg. Fluorescent microspheres were injected into the left ventricle. Regional ChBF was determined. All of the experimentally glaucomatous eyes exhibited supranormal first-order kernel (K1) root mean square (RMS) early portions of the mfERG waveforms and decreased amplitudes of the late waveforms. The supranormality was somewhat greater in the central macula. Second-order kernel, first slice (K2.1) RMS mfVEP response was inversely correlated (R(2) = 0.97) with axonal loss. Total ChBF was reduced in the experimentally glaucomatous eyes. The mean blood flow was 893 ± 123 and 481 ± 37 μL/min in the control and glaucomatous eyes, respectively. The ChBF showed regional variability with the greatest proportional decrement most often found in the central macula. This is the first demonstration of globally reduced ChBF in chronic experimental glaucoma in the nonhuman primate. Both the alteration of mfERG waveform components associated with outer retinal function and the reduction in ChBF were greatest in the macula, suggesting that there may be a spatial colocalization between ChBF and some outer retinal effects in glaucoma.


Purinergic receptors in skeletal muscles in health and in muscular dystrophy.
The P2 purinergic (nucleotide) receptor super-family comprises of two families of protein. The P2X, which are channel-forming ionotropic receptors and the P2Y metabotropic receptors activating G protein-mediated signalling pathways. Members of both groups have been identified in skeletal muscle cells at different stages of differentiation. It is well documented that sequential expression and down-regulation of particular P2 receptors on the surface of sarcolemma is closely associated with muscle maturation during embryogenesis and postnatal growth. P2 receptors are also involved in muscle regeneration following injury. Moreover, enhanced expression of specific purinergic receptors together with increased availability of extracellular ATP in dystrophic muscles are important elements of the dys- trophic pathophysiology considerably increasing severity.


Anti-agalactosyl IgG antibodies in sera from patients with systemic sclerosis.
To determine the prevalence and clinical correlations of anti-agalactosyl IgG antibodies (anti-AG IgG) in patients with systemic sclerosis (SSc). Serum samples from patients with limited cutaneous SSc (lSSc; n = 49), diffuse cutaneous SSc (dSSc; n = 21), rheumatoid arthritis (RA; n = 10), systemic lupus erythematosus (SLE; n = 20), and healthy individuals (n = 20) were examined by lectin-enzyme immunoassay using human agalactosyl IgG as antigen. Anti-AG IgG were detected in 52 (74%) of 70 patients with SSc, which was much higher than the frequency of rheumatoid factor positivity in SSc (16%). Levels of anti-agalactosyl IgG antibodies were significantly higher than in healthy controls or patients with SLE, but lower than patients with RA. Levels of anti-AG IgG in patients with dSSc were significantly higher than in lSSc. SSc patients with anti-topoisomerase I antibodies had significantly higher levels of anti-AG IgG than SSc patients with anticentromere antibodies. Concerning clinical correlation, patients with pulmonary fibrosis showed elevated levels of anti-AG IgG compared to those without pulmonary fibrosis. Patients with decreased %VC or %DLCO showed increased levels of anti-AG IgG. Elevated levels of anti-AG IgG were associated with the presence of contracture of phalanges or cutaneous calcinosis, but not the presence of arthritis/arthralgia. The results suggest that anti-agalactosyl IgG antibody is frequently detected in SSc and is a serological indicator for more severe SSc.


College student use of Salvia divinorum.
Salvia divinorum (salvia) is a plant that appears to be enjoying increased popularity as a legal hallucinogen in many U.S. jurisdictions. While the popular press has claimed that its use has become widespread, there have been no epidemiological studies published documenting this within the U.S. A sample of college students was randomly drawn from a large public university in the southwestern U.S. and invited to participate in an online survey that included salvia use among a set of other drug use items. From the sample of 1516 college student respondents, a pattern of use emerged that indicates that salvia is indeed becoming a significant member of the list of drugs used, with 4.4% of students reporting using salvia at least once within the past 12 months. Subpopulations that are typically most at risk for drug use within college students (Whites, males, fraternity members, heavy episodic drinkers) also were most likely to use salvia. The results indicate that more research is needed to determine the generalizability of these findings, and identify whether there are any negative consequences experienced either by the user or the community associated with this drug.


Automatic Selection of Integral Thresholds by Extrapolation in Coulomb and Exchange Matrix Constructions.
We present a method to compute Coulomb and exchange matrices with predetermined accuracy as measured by a matrix norm. The computation of these matrices is fundamental in Hartree-Fock and Kohn-Sham electronic structure calculations. We show numerically that, when modern algorithms for Coulomb and exchange matrix evaluation are applied, the Euclidean norm of the error matrix ε is related to the threshold value τ as ε = cτ(α). The presented extrapolation method automatically selects the integral thresholds so that the Euclidean norm of the error matrix is at the requested accuracy. This approach is demonstrated for a variety of systems, including protein-like systems, water clusters, and graphene sheets. The proposed method represents an important step toward complete error control throughout the self-consistent field calculation as described in [J. Math. Phys. 2008, 49, 032103].


Feasibility study of a field survey to measure antimicrobial usage in humans and animals in the Mekong Delta region of Vietnam.
Development of antimicrobial use (AMU) surveillance systems in humans and animals is a priority for many low- and middle-income countries; however accurate estimations are hampered by a diversity of animal production systems and metrics. The Mekong Delta region of Vietnam is a 'hotspot' of antimicrobial resistance and is home to a high density of humans and animal populations. To measure and compare AMU using different metrics (standing population, biomass and population correction unit) in the Mekong Delta, and to explore the potential of field-based data collection methods in the design of AMU surveillance systems. We collected AMU data from humans and animals (chickens, ducks, Muscovy ducks, pigs) from 101 small-scale farms in the Mekong Delta over a fixed period (90 days in humans, 7 days in animals). Humans used 7.1 DDDkg, or 175.9 mg of antimicrobial active ingredients (AAIs) per kg of standing body mass annually; animals consumed 60.9 ADDkg or 1324 mg. In the Mekong Delta humans represented 79.3% of the total body mass but consumed 29.6% of AAIs by weight. AAIs regarded of critical importance by WHO represented 56.9% and 50.2% of doses consumed by animals and humans, respectively. Using a One Health approach, we show that AMU can potentially be estimated from cross-sectional surveys, although results are hypothetical due to small sample size and are sensitive to the chosen population denominator. The methodology proposed here can potentially be scaled up be applied to design AMU surveillance in low-resource settings, allowing AMU reduction efforts to be focused on particular animal species.


A quantitative assessment of Arctic shipping in 2010-2014.
Rapid loss of sea ice is opening up the Arctic Ocean to shipping, a practice that is forecasted to increase rapidly by 2050 when many models predict that the Arctic Ocean will largely be free of ice toward the end of summer. These forecasts carry considerable uncertainty because Arctic shipping was previously considered too sparse to allow for adequate validation. Here, we provide quantitative evidence that the extent of Arctic shipping in the period 2011-2014 is already significant and that it is concentrated (i) in the Norwegian and Barents Seas, and (ii) predominantly accessed via the Northeast and Northwest Passages. Thick ice along the forecasted direct trans-Arctic route was still present in 2014, preventing transit. Although Arctic shipping remains constrained by the extent of ice coverage, during every September, this coverage is at a minimum, allowing the highest levels of shipping activity. Access to Arctic resources, particularly fisheries, is the most important driver of Arctic shipping thus far.


Preventive medical services not covered by public health insurance at Daiko Medical Center in Japan, 2004-2011.
Preventive medical services not covered by public health insurance started in the Daiko Medical Center of Nagoya University in June, 2004. Those services included: (1) Helicobacter pylori (H. pylori) diagnosis and eradication treatments, for which CYP2C19 genotyping was introduced in November 2005; (2) smoking cessation support with genotype tests of CYP1A1 Ile462Val, GSTM1 present/null, GSTT1 present/null, and NQO1 Pro187Ser; (3) advice on alcohol consumption with genotype tests of ADH Arg47His and ALDH2 Glu487Lys; (4) advice on folate-associated diseases with a genotype test of MTHFR C677T; (5) advice on a tumor marker CA19-9 with genotype tests of Lewis and Secretor genes; and (6) raloxifene prescription aimed to prevent breast cancer for high-risk postmenopausal women. A total of 683 patients visited the Center until it closed in March 2011. Those given diagnoses and eradication treatments for H. pylori numbered 567, followed by 44 for smoking cessation support, 35 for advice on folate-associated diseases, 26 for advice on alcohol consumption, 8 for CA19-9, and 3 for raloxifene prescription. Around 2004, public interest in H. pylori was relatively high, but thereafter patient numbers dropped markedly. The Center closed in March 2011 due to the reduction in patient visits. Our unique trial showed that continuing to provide uninsured preventive services at a clinic was difficult in Japan without the affiliation of hospitals/clinics providing medical services covered by public health insurance.


Enhanced detection of multiply phosphorylated peptides and identification of their sites of modification.
Phosphorylation is an important post-translational modification that rapidly mediates many cellular events. A key to understanding the dynamics of the phosphoproteome is localization of the modification site(s), primarily determined using LC-MS/MS. A major technical challenge to analysis is the formation of phosphopeptide-metal ion complexes during LC which hampers phosphopeptide detection. We have devised a strategy that enhances analysis of phosphopeptides, especially multiply phosphorylated peptides. It involves treatment of the LC system with EDTA and 2D-RP/RP-nanoUPLC-MS/MS (high pH/low pH) analysis. A standard triphosphorylated peptide that could not be detected with 1D-RP-nanoUPLC-MS/MS, even if the column was treated with EDTA-Na2 or if 25 mM EDTA-Na2 was added to the sample, was detectable at less than 100 fmol using EDTA-2D-RP/RP-nanoUPLC-MS/MS. Digests of α-casein and ß-casein were analyzed by EDTA-1D-RP-nanoUPLC, 2D-RP/RP-nanoUPLC, and EDTA-2D-RP/RP-nanoUPLC to compare their performance in phosphopeptide analysis. With the first two approaches, no tri- and tetraphosphopeptides were identified in either α- or ß-casein sample. With the EDTA-2D-RP/RP approach, 13 mono-, 6 di-, and 3 triphosphopeptides were identified in the α-casein sample, while 19 mono-, 8 di-, 4 tri-, and 3 tetraphosphopeptides were identified in the ß-casein sample. Using EDTA-2D-RP/RP-nanoUPLC-MS/MS to examine 500 μg of a human foreskin fibroblast cell lysate a total of 1,944 unique phosphopeptides from 1,087 unique phosphoproteins were identified, and 2,164 unique phosphorylation sites were confidently localized (Ascore ≥20). Of these sites 79% were mono-, 20% di-, and ∼1% were tri- and tetraphosphopeptides, and 78 novel phosphorylation sites in human proteins were identified.


Supplementary motor area and anterior intraparietal area integrate fine-graded timing and force control during precision grip.
We investigated the neuronal processing of the physiologically particularly important precision grip (opposition of index finger and thumb) by the combination of functional magnetic resonance imaging (fMRI) and an MR-compatible haptic interface. Ten healthy subjects performed isometric precision grip force generation with visual task instruction and real-time visual feedback in a block design. In a 2 x 2 two-factorial design, both the timing and force could be either constant or varying (identical average timing and force). As we expected only small changes in the fMRI response for the different fine-graded motor control conditions, we maximized the sensitivity of the data analysis and implemented a volumes of interest (VOI) restricted general linear model analysis including non-explanatory force regressors to eliminate directly force-related low-level activations. The VOIs were defined based on previous studies. We found significant associations: timing variation (variable vs. constant) and primary motor area (M1) and dorsal premotor area (PMd); force variation (variable vs. constant) and primary somatosensory area (S1), anterior intraparietal area (AIP) and PMd; interaction of timing and force and supplementary motor area (SMA) and AIP. We conclude that SMA and AIP integrate fine-graded higher-level timing and force control during precision grip. M1, S1 and PMd process lower-level timing and force control, yet not their integration. These results are the basis for a detailed assessment of manual motor control in a variety of motor diseases. The detailed behavioural assessment by our MR-compatible haptic interface is particularly valuable in patients due to expected larger inter-individual variation in motor performance.


Residential surrounding green, air pollution, traffic noise and self-perceived general health.
Self-perceived general health (SGH) is one of the most inclusive and widely used measures of health status and a powerful predictor of mortality. However, only a limited number of studies evaluated associations of combined environmental exposures on SGH. Our aim was to evaluate associations of combined residential exposure to surrounding green, air pollution and traffic noise with poor SGH in the Netherlands. We linked data on long-term residential exposure to surrounding green based on the Normalized Difference Vegetation Index (NDVI) and a land-use database (TOP10NL), air pollutant concentrations (including particulate matter (PM10, PM2.5), and nitrogen dioxide (NO2)) and road- and rail-traffic noise with a Dutch national health survey, resulting in a study population of 354,827 adults. We analyzed associations of single and combined exposures with poor SGH. In single-exposure models, NDVI within 300 m was inversely associated with poor SGH [odds ratio (OR) = 0.91, 95% CI: 0.89, 0.94 per IQR increase], while NO2 was positively associated with poor SGH (OR = 1.07, 95% CI: 1.04, 1.11 per IQR increase). In multi-exposure models, associations with surrounding green and air pollution generally remained, but attenuated. Joint odds ratios (JOR) of combined exposure to air pollution, rail-traffic noise and decreased surrounding green were higher than the odds ratios of single-exposure models. Studies including only one of these correlated exposures may overestimate the risk of poor SGH attributed to the studied exposure, while underestimating the risk of combined exposures.


Middle-term outcomes in renal transplant recipients with COVID-19: a national, multicenter, controlled study.
In this study, we evaluated 3-month clinical outcomes of kidney transplant recipients (KTR) recovering from COVID-19 and compared them with a control group. The primary endpoint was death in the third month. Secondary endpoints were ongoing respiratory symptoms, need for home oxygen therapy, rehospitalization for any reason, lower respiratory tract infection, urinary tract infection, biopsy-proven acute rejection, venous/arterial thromboembolic event, cytomegalovirus (CMV) infection/disease and BK viruria/viremia at 3 months. A total of 944 KTR from 29 different centers were included in this study (523 patients in the COVID-19 group; 421 patients in the control group). The mean age was 46 ± 12 years (interquartile range 37-55) and 532 (56.4%) of them were male. Total number of deaths was 8 [7 (1.3%) in COVID-19 group, 1 (0.2%) in control group; P = 0.082]. The proportion of patients with ongoing respiratory symptoms [43 (8.2%) versus 4 (1.0%); P < 0.001] was statistically significantly higher in the COVID-19 group compared with the control group. There was no significant difference between the two groups in terms of other secondary endpoints. The prevalence of ongoing respiratory symptoms increased in the first 3 months post-COVID in KTRs who have recovered from COVID-19, but mortality was not significantly different.


Testing the assumptions for the analysis of survival data arising from a prevalent cohort study with follow-up.
In a prevalent cohort study with follow-up subjects identified as prevalent cases are followed until failure (defined suitably) or censoring. When the dates of the initiating events of these prevalent cases are ascertainable, each observed datum point consists of a backward recurrence time and a possibly censored forward recurrence time. Their sum is well known to be the left truncated lifetime. It is common to term these left truncated lifetimes "length biased" if the initiating event times of all the incident cases (including those not observed through the prevalent sampling scheme) follow a stationary Poisson process. Statistical inference is then said to be carried out under stationarity. Whether or not stationarity holds, a further assumption needed for estimation of the incident survivor function is the independence of the lifetimes and their accompanying truncation times. That is, it must be assumed that survival does not depend on the calendar date of the initiating event. We show how this assumption may be checked under stationarity, even though only the backward recurrence times and their associated (possibly censored) forward recurrence times are observed. We prove that independence of the lifetimes and truncation times is equivalent to equality in distribution of the backward and forward recurrence times, and exploit this equivalence as a means of testing the former hypothesis. A simulation study is conducted to investigate the power and Type 1 error rate of our proposed tests, which include a bootstrap procedure that takes into account the pairwise dependence between the forward and backward recurrence times, as well as the potential censoring of only one of the members of each pair. We illustrate our methods using data from the Canadian Study of Health and Aging. We also point out an equivalence of the problem presented here to a non-standard changepoint problem.


Epidermal growth factor receptor (EGFR) ubiquitination as a mechanism of acquired resistance escaping treatment by the anti-EGFR monoclonal antibody cetuximab.
Cetuximab is an epidermal growth factor receptor (EGFR)-blocking antibody that has been approved for treatment of patients with metastatic colorectal cancer. In this study, we investigated biochemical changes in signaling pathways of a cetuximab-resistant subline of DiFi colorectal cancer cells (DiFi5) that was developed by exposing the parental sensitive cells to subeffective doses of cetuximab over an extended period of time. Compared with parental DiFi cells that express high levels of EGFR and in which cetuximab induces apoptosis, the cetuximab-resistant DiFi5 cells showed markedly lower protein levels of EGFR, an increased association of EGFR with Cbl, and an increased ubiquitination of EGFR. DiFi5 cells also had a markedly higher level of Src-Y416 phosphorylation both at baseline and on EGF stimulation. Although EGFR levels were low, DiFi5 cells responded to EGF stimulation with robust phosphorylation of EGFR on Y845 and strong phosphorylation of Akt and extracellular signal-regulated kinase, comparable to those of parental cells. Most importantly, inhibition of Src kinase activity with PP2 reversed the resistance of DiFi5 cells to cetuximab-induced apoptosis without affecting the levels of EGFR in the cells. Our results indicate that colorectal cancer cells may develop acquired resistance to cetuximab via altering EGFR levels through promotion of EGFR ubiquitination and degradation and using Src kinase-mediated cell signaling to bypass their dependency on EGFR for cell growth and survival.


[Health surveillance of workers in indoor environments. Application of the Italian version of the MM040/IAQ questionnaire].
To test the efficacy of the Italian version of a questionnaire used for the health surveillance of workers operating in confined spaces. The Italian version of the Swedish MM040/Indoor Air Questionnaire was used to collect information on ailments and symptoms related to the indoor environment of 28 workplaces. 4,029 workers took part in the study. For 23.1% of respondents, the most frequent environmental problem concerned fluctuations in temperature. For 12.7% of respondents, the commonest work-related symptom was fatigue. Women reported indoor air problems and work-related symptoms more often than men. Allergic subjects reported environmental problems and work-related symptoms more often than non-allergic workers. A significant correlation was found between environmental discomfort and the presence of symptoms. Ailments and work-related symptoms were common in indoor workers, regardless of the presence of collective disorders such as the "sick building syndrome" (SBS). The questionnaire used in this study proved valuable in helping the occupational health physician to monitor the health of indoor workers. The data collected can provide a reference point for the medical surveillance of workers operating in confined spaces.


Clinical Epidemiology of Septic Arthritis Caused by Burkholderia pseudomallei and Other Bacterial Pathogens in Northeast Thailand.
Septic arthritis is a medical emergency, and if not treated appropriately, it can be associated with high morbidity and mortality. Melioidosis, a serious infectious disease caused by the Gram-negative bacillus Burkholderia pseudomallei, is highly endemic in South and Southeast Asia and northern Australia. We reviewed the medical charts of adult patients admitted with bacterial septic arthritis at Sunpasitthiprasong Hospital, Ubon Ratchathani, northeast Thailand from January 2012 to December 2014. Bacterial septic arthritis was defined as one or more hot swollen joints with isolation of a pathogenic organism from an affected joint or from blood. A total of 154 patients with septic arthritis were retrospectively evaluated. The most common causes were B. pseudomallei (48%, N = 74), Streptococcus spp. (29%, N = 44), and Staphylococcus aureus (10%, N = 16). Prevalence of diabetes, bacteremia, and pneumonia was higher in B. pseudomallei septic arthritis than in septic arthritis caused by the other bacteria (all P < 0.01). Seventy three percent (54/74) of patients infected with B. pseudomallei and 69% (55/80) of patients with the other bacteria received effective antimicrobials on the first day of admission (P = 0.60), but in-hospital mortality of the former group was considerably higher (34% versus 14%, P = 0.004). In conclusion, B. pseudomallei septic arthritis is common and associated with high mortality in northeast Thailand. Emergence of Streptococcus arthritis is observed. Difficulty in diagnosing melioidosis and identifying B. pseudomallei in areas where health care workers are not familiar with the disease is discussed. In melioidosis-endemic regions, parenteral ceftazidime could be considered as empirical antimicrobial therapy for patients with septic arthritis and underlying diseases.


Effect of 2.45 GHz microwave radiation on permeability of unilamellar liposomes to 5(6)-carboxyfluorescein. Evidence of non-thermal leakage.
The influence of 2.45 GHz microwave radiation on the membrane permeability of unilamellar liposomes was studied using the marker 5(6)-carboxyfluorescein trapped in phosphatidylcholine liposomes. The release of the fluorescent marker was followed by spectrofluorimetry after an exposure of 10 minutes to either microwave radiation or to heat alone of the liposome solutions. A significant increase of the permeability of carboxyfluorescein through the membrane was observed for the microwave-exposed samples compared to those exposed to normal heating only. Exposure to 2.45 GHz microwave radiation of liposomes has been previously found to produce increased membrane permeability as compared with heating. However, in contrast to previous studies, the observations reported here were made above the phase transition temperature of the lipid membrane. The experimental setup included monitoring of the temperature during microwave exposure simultaneously at several points in the solution volume using a fiberoptic thermometer. Possible mechanisms to explain the observations are discussed.


Cannabis use in pediatric cancer patients: what are they reading? A review of the online literature.
Recent changes to the legal status of marijuana in Canada warrant a review of the information that patients and families are accessing online regarding the role of cannabis in cancer. The aims of the current research were to identify the quality of literature available online as well as the themes, and opinion (i.e., pro-, neutral, or anti-cannabis) of online articles. Searches were conducted using three primary search engines: Google, Yahoo, and DuckDuckGo. Articles were assessed for quality based on a modified scale for evaluating online sources. Content of all unique articles was coded using a qualitative thematic methodology in a line-by-line fashion. Codes were clustered to determine themes within articles. Finally, opinions were determined by examining all articles in a line-by-line fashion. Each statement was coded as either pro-cannabis (positive) or anti-cannabis (negative). We found most articles were authored by journalists (39.4%) and MDs (14.1%) and published as news (35.2%) or web articles (28.2%). The content of articles focused on four themes: the reasons for and against cannabis use; the opinions of health care providers; the restrictions placed by governing bodies and the need for additional research, education, and standardization. Article opinions were neutral-pro-cannabis. Health care providers should be aware that the overall quality of information found online is considered "satisfactory." The majority of articles present a pro-cannabis opinion.


Management of Root Resorption Using Chemical Agents: A Review.
Root resorption (RR) is defined as the loss of dental hard tissues because of clastic activity inside or outside of tooth the root. In the permanent dentition, RR is a pathologic event; if untreated, it might result in the premature loss of the affected tooth. Several hypotheses have been suggested as the mechanisms of root resorption such as absence of the remnants of Hertwig's epithelial root sheath (HERS) and the absence of some intrinsic factors in cementum and predentin such as amelogenin or osteoprotegerin (OPG). It seems that a barrier is formed by the less-calcified intermediate cementum or the cementodentin junction that prevents external RR. There are several chemical strategies to manage root resorption. The purpose of this paper was to review several chemical agents to manage RR such as tetracycline, sodium hypochlorite, acids (citric acid, phosphoric acid, ascorbic acid and hydrochloric acid), acetazolamide, calcitonin, alendronate, fluoride, Ledermix and Emdogain.


Results of a randomized controlled trial on statin use in dialysis patients had no influence on statin prescription.
Randomized trials provide high-quality evidence for patient care. The Der Deutsche Diabetes Dialyse Studie (4D), a randomized study which demonstrated no benefit of statins among diabetic patients receiving hemodialysis, was published in July 2005. To determine effects of this study we conducted a retrospective, population-based, time series analysis with change-point regression to see if the rate of statin prescription to dialysis patients had been modified. We linked health administrative data for all diabetic hemodialysis patients living in Ontario, Canada, with similar characteristics to the 4D patient cohort. During the nearly 11-year period prior to study publication, the rate of statin use increased almost 14-fold, from 43 to 597 per 1000 patients. For 2.5 years after study publication, rather than diminish, statin use continued to rise to an absolute rate of 676 per 1000 patients. These temporal patterns in statin use closely mimicked trends in the diabetic population not receiving dialysis. The 4D trial had no impact on statin use when we restricted the analysis to incident statin prescriptions or expanded the characteristics of the dialysis patients considered for study. Thus, we found that publication of a large, expensive, randomized controlled trial in patients receiving hemodialysis had no immediate impact on clinical practice. The use of a common cardiovascular medication in this patient population appears to be influenced by other factors.


Correlates of health-related quality of life among adults receiving combination antiretroviral therapy in coastal Kenya.
Health-related quality of life (HRQoL) is an important metric of perceived wellbeing in people living with HIV/AIDS (PLWHA). However, research on HRQoL among PLWHA in sub-Saharan Africa is limited. This study investigates factors associated with HRQoL among PLWHA in Kilifi, coast of Kenya. Between February and April 2018, 450 adults living with HIV and on combined antiretroviral therapy (cART) between 18 to 60 years were sequentially recruited from an HIV-specialized clinic. The Functional Assessment of HIV Infection (FAHI) questionnaire, previously adapted for assessing HRQoL in this setting, was slightly modified and administered to participants alongside other measures of sociodemographic, health and treatment characteristics in a face-to-face interview. Linear regression analyses indicated that depressive symptoms, HIV-related stigma, non-disclosure of HIV status, living alone, clinic inaccessibility, and presence of any current opportunistic infection were significantly associated with lower HRQoL scores at both the FAHI overall and sub-scale level. Higher physician empathy, male sex, and higher body mass index were significantly associated with better HRQoL scores at both FAHI overall and sub-scale level. Age and longer duration on cART were significantly associated with better HRQoL only at the sub-scale level. Interventions aimed at reducing depressive symptoms and HIV stigma, making HIV-related services more accessible, addressing opportunistic infections, strengthening social support systems, serostatus disclosure and put in place caring, respectful, and compassionate model of care are necessary to improve the HRQoL of PLWHA.


pHuji, a pH-sensitive red fluorescent protein for imaging of exo- and endocytosis.
Fluorescent proteins with pH-sensitive fluorescence are valuable tools for the imaging of exocytosis and endocytosis. The Aequorea green fluorescent protein mutant superecliptic pHluorin (SEP) is particularly well suited to these applications. Here we describe pHuji, a red fluorescent protein with a pH sensitivity that approaches that of SEP, making it amenable for detection of single exocytosis and endocytosis events. To demonstrate the utility of the pHuji plus SEP pair, we perform simultaneous two-color imaging of clathrin-mediated internalization of both the transferrin receptor and the β2 adrenergic receptor. These experiments reveal that the two receptors are differentially sorted at the time of endocytic vesicle formation.


Impact of subclinical hypothyroidism on the coronary artery disease in apparently healthy subjects.
Cardiovascular disease (CVD) occurs frequently and may progress more rapidly in overt hypothyroidism (OVH). However, the role of mild thyroid failure as a risk factor for CVD is not clear. This study is aimed at exploring the association between subclinical hypothyroidism (SCH) and coronary artery disease (CAD), as detected by cardiac computed tomography (CT), in apparently healthy subjects. We retrospectively enrolled 2404 asymptomatic subjects who underwent cardiac CT with an intermediate to high risk (Framingham 10-year risk ≥10%) of developing CAD but with no known CAD or thyroid disease. Coronary artery calcium score (CACS) was assessed by calcium scan, and the presence of the plaques (CAD), with ≥50% stenosis being indicative of obstructive CAD, was assessed by coronary CT angiography. Of the 2404 subjects, 2355 subjects were euthyroid (Eu; 53±9 years, 83 females) and 49 had SCH (58±12 years, seven females). CAD and CACS >100 were more prevalent in SCH subjects than in Eu subjects (Eu vs SCH: CAD, 948 (40.6%) vs 31 (63.3%), P=0.002; CACS >100, 239 (10.3%) vs 10 (20.4%), P=0.031). SCH was also an independent risk factor for CAD after a multivariate analysis (odds ratio: 2.125, 95% confidence interval: 1.049-4.307, P=0.036). SCH subjects who were at an intermediate-to-high risk of developing CAD were significantly more likely to exhibit occult CAD than Eu subjects, especially in men with SCH. These findings suggest that mild thyroid failure also independently contributes to the development of CAD.


TRAP transporters: a new family of periplasmic solute transport systems encoded by the dctPQM genes of Rhodobacter capsulatus and by homologs in diverse gram-negative bacteria.
The dct locus of Rhodobacter capsulatus encodes a high-affinity transport system for the C4-dicarboxylates malate, succinate, and fumarate. The nucleotide sequence of the region downstream of the previously sequenced dctP gene (encoding a periplasmic C4-dicarboxylate-binding protein) was determined. Two open reading frames (ORFs) of 681 bp (dctQ) and 1,320 bp (dctM) were identified as additional dct genes by insertional mutagenesis and complementation studies. DctQ (24,763 Da) and DctM (46,827 Da) had hydropathic profiles consistent with the presence of 4 and 12 potential transmembrane segments, respectively, and were localized in the cytoplasmic membrane fraction after heterologous expression of the dctQM ORFs in Escherichia coli. DctP, DctQ, and DctM were found to be unrelated to known transport proteins in the ABC (ATP-binding cassette) superfamily but were shown to be homologous with the products of previously unidentified ORFs in a number of gram-negative bacteria, including Bordetella pertussis, E. coli, Salmonella typhimurium, Haemophilus influenzae, and Synechocystis sp. strain PCC6803. An additional ORF (rypA) downstream of dctM encodes a protein with sequence similarity to eukaryotic protein-tyrosine phosphatases, but interposon mutagenesis of this ORF did not result in a Dct- phenotype. Complementation of a Rhizobium meliloti dctABD deletion mutant by heterologous expression of the dctPQM genes from R. capsulatus demonstrated that no additional structural genes were required to form a functional transport system. Transport via the Dct system was vanadate insensitive, and in uncoupler titrations with intact cells, the decrease in the rate of succinate transport correlated closely with the fall in membrane potential but not with the cellular ATP concentration, implying that the proton motive force, rather than ATP hydrolysis, drives uptake. It is concluded that the R. capsulatus Dct system is a new type of periplasmic secondary transporter and that similar, hitherto-unrecognized systems are widespread in gram-negative bacteria. The name TRAP (for tripartite ATP-independent periplasmic) transporters is proposed for this new group.


De novo variants in CDK13 associated with syndromic ID/DD: Molecular and clinical delineation of 15 individuals and a further review.
De novo variants in the gene encoding cyclin-dependent kinase 13 (CDK13) have been associated with congenital heart defects and intellectual disability (ID). Here, we present the clinical assessment of 15 individuals and report novel de novo missense variants within the kinase domain of CDK13. Furthermore, we describe 2 nonsense variants and a recurrent frame-shift variant. We demonstrate the synthesis of 2 aberrant CDK13 transcripts in lymphoblastoid cells from an individual with a splice-site variant. Clinical characteristics of the individuals include mild to severe ID, developmental delay, behavioral problems, (neonatal) hypotonia and a variety of facial dysmorphism. Congenital heart defects were present in 2 individuals of the current cohort, but in at least 42% of all known individuals. An overview of all published cases is provided and does not demonstrate an obvious genotype-phenotype correlation, although 2 individuals harboring a stop codons at the end of the kinase domain might have a milder phenotype. Overall, there seems not to be a clinically recognizable facial appearance. The variability in the phenotypes impedes an à vue diagnosis of this syndrome and therefore genome-wide or gene-panel driven genetic testing is needed. Based on this overview, we provide suggestions for clinical work-up and management of this recently described ID syndrome.


Robust finger vein ROI localization based on flexible segmentation.
Finger veins have been proved to be an effective biometric for personal identification in the recent years. However, finger vein images are easily affected by influences such as image translation, orientation, scale, scattering, finger structure, complicated background, uneven illumination, and collection posture. All these factors may contribute to inaccurate region of interest (ROI) definition, and so degrade the performance of finger vein identification system. To improve this problem, in this paper, we propose a finger vein ROI localization method that has high effectiveness and robustness against the above factors. The proposed method consists of a set of steps to localize ROIs accurately, namely segmentation, orientation correction, and ROI detection. Accurate finger region segmentation and correct calculated orientation can support each other to produce higher accuracy in localizing ROIs. Extensive experiments have been performed on the finger vein image database, MMCBNU_6000, to verify the robustness of the proposed method. The proposed method shows the segmentation accuracy of 100%. Furthermore, the average processing time of the proposed method is 22 ms for an acquired image, which satisfies the criterion of a real-time finger vein identification system.


Selenoneine ameliorates peroxide-induced oxidative stress in C. elegans.
Selenoneine (2-selenyl-Nα, Nα, Nα-trimethyl-L-histidine), the selenium (Se) analogue of the ubiquitous thiol compound and putative antioxidant ergothioneine, is the major organic selenium species in several marine fish species. Although its antioxidant efficacy has been proposed, selenoneine has been poorly characterized, preventing conclusions on its possible beneficial health effects. Treatment of Caenorhabditis elegans (C. elegans) with selenoneine for 18 h attenuated the induction of reactive oxygen and nitrogen species (RONS). However, the effect was not immediate, occurring 48 h post-treatment. Total Se and Se speciation analysis revealed that selenoneine was efficiently taken up and present in its original form directly after treatment, with no metabolic transformations observed. 48 h post-treatment, total Se in worms was slightly higher compared to controls and no selenoneine could be detected. The protective effect of selenoneine may not be attributed to the presence of the compound itself, but rather to the activation of molecular mechanisms with consequences at more protracted time points.


Multi-frame super-resolution with quality self-assessment for retinal fundus videos.
This paper proposes a novel super-resolution framework to reconstruct high-resolution fundus images from multiple low-resolution video frames in retinal fundus imaging. Natural eye movements during an examination are used as a cue for super-resolution in a robust maximum a-posteriori scheme. In order to compensate heterogeneous illumination on the fundus, we integrate retrospective illumination correction for photometric registration to the underlying imaging model. Our method utilizes quality self-assessment to provide objective quality scores for reconstructed images as well as to select regularization parameters automatically. In our evaluation on real data acquired from six human subjects with a low-cost video camera, the proposed method achieved considerable enhancements of low-resolution frames and improved noise and sharpness characteristics by 74%. In terms of image analysis, we demonstrate the importance of our method for the improvement of automatic blood vessel segmentation as an example application, where the sensitivity was increased by 13% using super-resolution reconstruction.


Moyamoya disease: a clinical spectrum, literature review and case series from a tertiary care hospital in Pakistan.
Moyamoya is a rare cerebrovascular disease of unknown etiology. The data on moyamoya disease from Pakistan is sparse. We report a case series of 13 patients who presented with moyamoya disease to a tertiary care hospital in Pakistan with a national referral base. We conducted a retrospective review of thirteen patients who presented to The Aga Khan University and diagnosed with "Moyamoya Disease" during the period 1988 - 2006. These patients were identified from existing hospital database via ICD-9 codes. A predesigned questionnaire containing information about clinical presentation, management and neuroimaging was administered to all identified patients. There were seven males and six females. Mean age at presentation was 16.5 years and a female predominance was found in the pediatric age group (n = 10, 71.4%). Stroke (n = 11, 84.2%) was the most common presentation with motor deficit being the universal cortical symptom. Fever was a common symptom in the lower age groups (n = 4, 51.7%). Cerebral Angiography and Magnetic Resonance Angiography showed bilateral involvement of the vessels in eleven patients while unilateral in two. Subarachnoid and interventricular haemorrhage appeared in 2(15.4%) adults. Twelve (92.3%) patients were discharged as independent with minor deficits regardless of therapeutic modality. Only three (23.0%) patients underwent surgery whereas the remaining were managed conservatively. Physicians when dealing with childhood strokes and characteristic deficits in adult population should consider Moyamoya disease.


Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus.
A stable deoxyribonucleic acid (DNA) polymerase (EC 126.96.36.199) with a temperature optimum of 80 degrees C has been purified from the extreme thermophile Thermus aquaticus. The enzyme is free from phosphomonoesterase, phosphodiesterase and single-stranded exonuclease activities. Maximal activity of the enzyme requires all four deoxyribonucleotides and activated calf thymus DNA. An absolute requirement for divalent cation cofactor was satisfied by Mg2+ or to a lesser extent by Mn2+. Monovalent cations at concentrations as high as 0.1 M did not show a significant inhibitory effect. The pH optimum was 8.0 in tris(hydroxymethyl)aminomethane-hydrochloride buffer. The molecular weight of the enzyme was estimated by sucrose gradient centrifugation and gel filtrations on Sephadex G-100 to be approximately 63,000 to 68,000. The elevated temperature requirement, small size, and lack of nuclease activity distinguish this polymerase from the DNA polymerase of Escherichia coli.


A Voxel-by-Voxel Comparison of Deformable Vector Fields Obtained by Three Deformable Image Registration Algorithms Applied to 4DCT Lung Studies.
Commonly used methods of assessing the accuracy of deformable image registration (DIR) rely on image segmentation or landmark selection. These methods are very labor intensive and thus limited to relatively small number of image pairs. The direct voxel-by-voxel comparison can be automated to examine fluctuations in DIR quality on a long series of image pairs. A voxel-by-voxel comparison of three DIR algorithms applied to lung patients is presented. Registrations are compared by comparing volume histograms formed both with individual DIR maps and with a voxel-by-voxel subtraction of the two maps. When two DIR maps agree one concludes that both maps are interchangeable in treatment planning applications, though one cannot conclude that either one agrees with the ground truth. If two DIR maps significantly disagree one concludes that at least one of the maps deviates from the ground truth. We use the method to compare 3 DIR algorithms applied to peak inhale-peak exhale registrations of 4DFBCT data obtained from 13 patients. All three algorithms appear to be nearly equivalent when compared using DICE similarity coefficients. A comparison based on Jacobian volume histograms shows that all three algorithms measure changes in total volume of the lungs with reasonable accuracy, but show large differences in the variance of Jacobian distribution on contoured structures. Analysis of voxel-by-voxel subtraction of DIR maps shows differences between algorithms that exceed a centimeter for some registrations. Deformation maps produced by DIR algorithms must be treated as mathematical approximations of physical tissue deformation that are not self-consistent and may thus be useful only in applications for which they have been specifically validated. The three algorithms tested in this work perform fairly robustly for the task of contour propagation, but produce potentially unreliable results for the task of DVH accumulation or measurement of local volume change. Performance of DIR algorithms varies significantly from one image pair to the next hence validation efforts, which are exhaustive but performed on a small number of image pairs may not reflect the performance of the same algorithm in practical clinical situations. Such efforts should be supplemented by validation based on a longer series of images of clinical quality.


Characterisation of PD-L1-positive subsets of microsatellite-unstable colorectal cancers.
The aim of this study was to reveal the clinicopathological and molecular characteristics of microsatellite instability-high (MSI-H) colorectal cancers (CRCs) showing programmed death ligand-1 (PD-L1) positivity, which are good candidates for anti-PD-1/PD-L1 immunotherapy. The PD-L1 expression status of 208 MSI-H CRCs was retrospectively analysed using immunohistochemistry. PD-L1 positivity in tumour cells (PD-L1+(T)) and PD-L1 positivity in immune cells (PD-L1+(I)) were separately evaluated. Programmed death ligand-1 positivity in tumour cells and PD-L1+(I) were observed in 26 (12.5%) and 62 (29.8%) MSI-H CRCs, respectively, and occasionally overlapped (n=12; 5.8%). Programmed death ligand-1 positivity tumours in MSI-H CRCs were significantly associated with older age, female sex, non-mucinous-type poor differentiation, infiltrating growth, tumour budding, advanced stage, CpG island methylator phenotype-high, MLH1 promoter methylation, and BRAF V600E mutations. However, PD-L1+(I) MSI-H CRCs were characterised by high-density tumour-infiltrating immune cells, including T cells and macrophages, and intense peritumoural lymphoid reactions. In patients with stage IV MSI-H CRCs who had undergone metastatectomy (n=4), the PD-L1 status of primary tumours was maintained in corresponding distant metastatic lesions. In MSI-H CRCs, PD-L1+(T) and PD-L1+(I) are linked to a sporadic hypermethylated subtype and an immune cell-rich subtype, respectively. Potential differential therapeutic implications of PD-L1+(T) and PD-L1+(I) in CRCs should be further investigated.


Continuing observations on the regulatory effects of donor-specific bone marrow cell infusions and chimerism in kidney transplant recipients.
Continued follow-up of a series of donor bone marrow cell (DBMC)-infused first cadaver renal transplant recipients is described (n=58), now at a 36-month actuarial time point postoperatively. Serial polymerase chain reaction-flow cytometry (PCR-Flow) and cellular immune assays of iliac crest bone marrow aspirates and peripheral blood have begun to be compared with concomitantly transplanted recipients of living-related donor (LRD) kidneys and donor marrow infusions given the same immunosuppressive regimen (n=16). There have also been comparisons (36 months) with 188 controls transplanted concomitantly, i.e., recipients of first cadaver kidney transplants, who did not receive bone marrow. Each group was given equivalent immunosuppressive regimens of OKT3 anti-T cell induction and maintenance tacrolimus, mycophenolate mofetil, and methylprednisolone. Actuarial patient and graft survival have been 96% and 93%, respectively, in the controls and 91% and 91%, respectively, in the DBMC-infused recipients. Trough levels of tacrolimus were significantly lower in the DBMC-infused group. In PCR-Flow measurements, in peripheral blood up to 6 months postoperatively, there were higher levels of chimerism, i.e., in the total number of donor cells, as well as the donor CD3+ and CD34+ subsets in the LRD recipients administered DBMC infusions, compared with cadaver DBMC recipients, supporting the notion of a positive effect of histocompatibility on chimerism levels. In PCR-Flow measurements of recipient iliac crest bone marrow aspirates as in previous studies on peripheral blood, early acute rejection episodes (<1 month) were found to be associated with a later (6-14 months) decrease in donor cell lineage chimerism. However, a trend toward recovery of chimeric levels occurred by 21-28 months in a second iliac crest marrow aspirate 1 year after the first aspirate in the DBMC-infused recipients who experienced such early rejection episodes. This was in contrast to the controls in whom there were sustained low levels of iliac crest bone marrow chimerism at both the earlier and later intervals (i.e., no chimeric recovery), with 17/183 surviving controls progressing into chronic rejection. This has not yet been seen in the DBMC-infused group (0/54). In in vitro observations on cellular immune reactivity at 1 year postoperatively, decreased peripheral blood lymphocyte proliferative reactions were seen in response to phytohemagglutinin and Staph-A mitogens, as well as to cytomegalovirus and Epstein-Barr viral protein antigens in the DBMC-infused group versus the controls. Chronic immunosuppression did not seem to effect a vigorous in vitro inhibitory (regulatory) activity of bone marrow taken from these transplant recipients 2 years postoperatively in mixed lymphocyte culture and cell-mediated lympholysis reactions, using allogeneic responding cells from "normal" laboratory volunteers. Autologous peripheral blood lymphoproliferative responses to phytohemagglutinin and Staph-A mitogens, as well as to cytomegalovirus and Epstein-Barr virus protein antigens, were also regulated by either organ donor (non-immunosuppressed) bone marrow cells or by transplant recipient (immunosuppressed) bone marrow cells. What appeared to be disparate between the DBMC-infused and control groups (both immunosuppressed) was the trend for the (autologous) bone marrow suppressive effect on antiviral lymphoproliferative responses, to be stronger in the DBMC-infused group, who also had significantly (>one order of magnitude) higher levels of chimerism (P=0.01). It is concluded that the establishment of a chimeric state in DBMC-infused recipients, albeit of relatively low magnitude (approximately 1% at 2 years in recipient iliac crest bone marrow), has had a definite regulatory effect on immune responses. These results, therefore, add weight to the "causal" horn of the dilemma as to whether donor cell chimerism is a cause or an effect of


Fine-Grained 3D-Attention Prototypes for Few-Shot Learning.
In the real world, a limited number of labeled finely grained images per class can hardly represent the class distribution effectively. Due to the more subtle visual differences in fine-grained images than simple images with obvious objects, that is, there exist smaller interclass and larger intraclass variations. To solve these issues, we propose an end-to-end attention-based model for fine-grained few-shot image classification (AFG) with the recent episode training strategy. It is composed mainly of a feature learning module, an image reconstruction module, and a label distribution module. The feature learning module mainly devises a 3D-Attention mechanism, which considers both the spatial positions and different channel attentions of the image features, in order to learn more discriminative local features to better represent the class distribution. The image reconstruction module calculates the mappings between local features and the original images. It is constrained by a designed loss function as auxiliary supervised information, so that the learning of each local feature does not need extra annotations. The label distribution module is used to predict the label distribution of a given unlabeled sample, and we use the local features to represent the image features for classification. By conducting comprehensive experiments on Mini-ImageNet and three fine-grained data sets, we demonstrate that the proposed model achieves superior performance over the competitors.


Primary structure of a beta subunit of alpha-dendrotoxin-sensitive K+ channels from bovine brain.
Voltage-dependent cation channels are large heterooligomeric proteins. Heterologous expression of cDNAs encoding the alpha subunits alone of K+, Na+, or Ca2+ channels produces functional multimeric proteins; however, coexpression of those for the latter two with their auxiliary proteins causes dramatic changes in the resultant membrane currents. Fast-activating, voltage-sensitive K+ channels from brain contain four alpha and beta subunits, tightly associated in a 400-kDa complex; although molecular details of the alpha-subunit proteins have been determined, little is known about the beta-subunit constituent. Proteolytic fragments of a beta subunit from bovine alpha-dendrotoxin-sensitive neuronal K+ channels yielded nine different sequences. In the polymerase chain reaction, primers corresponding to two of these peptides amplified a 329-base-pair fragment in a lambda gt10 cDNA library from bovine brain; a full-length clone subsequently isolated encodes a protein of 367 amino acids (M(r) approximately 40,983). It shows no significant homology with any known protein. Unlike the channels' alpha subunits, the hydropathy profile of this sequence failed to reveal transmembrane domains. Several consensus phosphorylation motifs are apparent and, accordingly, the beta subunit could be phosphorylated in the intact K+ channels. These results, including the absence of a leader sequence and N-glycosylation, are consistent with the beta subunit being firmly associated on the inside of the membrane with alpha subunits, as speculated in a simplified model of these authentic K+ channels. Importantly, this first primary structure of a K(+)-channel beta subunit indicates that none of the cloned auxiliary proteins of voltage-dependent cation channels, unlike their alpha subunits, belong to a super-family of genes.


Uncertainty analysis for common Seebeck and electrical resistivity measurement systems.
This work establishes the level of uncertainty for electrical measurements commonly made on thermoelectric samples. The analysis targets measurement systems based on the four probe method. Sources of uncertainty for both electrical resistivity and Seebeck coefficient were identified and evaluated. Included are reasonable estimates on the magnitude of each source, and cumulative propagation of error. Uncertainty for the Seebeck coefficient includes the cold-finger effect which has been quantified with thermal finite element analysis. The cold-finger effect, which is a result of parasitic heat transfer down the thermocouple probes, leads to an asymmetric over-estimation of the Seebeck coefficient. A silicon germanium thermoelectric sample has been characterized to provide an understanding of the total measurement uncertainty. The electrical resistivity was determined to contain uncertainty of ±7.0% across any measurement temperature. The Seebeck coefficient of the system is +1.0%/-13.1% at high temperature and ±1.0% near room temperature. The power factor has a combined uncertainty of +7.3%/-27.0% at high temperature and ±7.5% near room temperature. These ranges are calculated to be typical values for a general four probe Seebeck and resistivity measurement configuration.


The early landscape of coronavirus disease 2019 vaccine development in the UK and rest of the world.
Since the first World Health Organization notification on 31 December 2019, coronavirus disease 2019 (COVID-19), the respiratory disease caused by the coronavirus severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), has been responsible for over four million confirmed infections and almost 300 000 deaths worldwide. The pandemic has led to over half of the world's population living under lockdown conditions. To allow normal life to resume, public health interventions will be needed to prevent further waves of infections as lockdown measures are lifted. As one of the most effective countermeasures against infectious diseases, an efficacious vaccine is considered crucial to containing the COVID-19 pandemic. Following the publication of the genome sequence of SARS-CoV-2, vaccine development has accelerated at an unprecedented pace across the world. Here we review the different platforms employed to develop vaccines, the standard timelines of development and how they can be condensed in a pandemic situation. We focus on vaccine development in the UK and vaccines that have entered clinical trials around the world.


The effects of pre-meal drink volume on gastric motility and energy intake in healthy men.
Pre-meal drink ingestion is an effective method of controlling energy intake in humans. However, no studies have addressed the influence of differences in the volume of drink intake on gastric motility and energy intake. The purpose of the present study was to examine the effects of differences in the volume of drink intake before a meal on subsequent gastric motility and energy intake in healthy young men. Twelve men completed two, one-day trials in a random order. Subjects visited the laboratory after a 10-h overnight fast and consumed the nutrient drink (0.84 MJ) in either 100 mL or 600 mL quantities over a 5-min period. Then, the subjects sat on a chair for over 2 h to measure their cross-sectional gastric antral areas and gastric contractions with an ultrasound imaging system. Thereafter, the subjects consumed a test meal until they felt completely full. Energy intake was calculated from the amount of food consumed. Energy intake in the 600 mL trial was 12% higher than the 100 mL trial (5.1 ± 1.3 vs. 4.6 ± 1.4 MJ, P = 0.046). The antral area (P = 0.046) and the frequency of the gastric contraction (P = 0.001) over 2 h after consuming the nutrient drink were higher in the 600 mL trial than the 100 mL trial. These findings demonstrated that consumption of a 600 mL nutrient drink increased energy intake. The modulation of gastric motility might have some effects on energy intake.


Blood, urine, and sweat (BUS) study: monitoring and elimination of bioaccumulated toxic elements.
There is limited understanding of the toxicokinetics of bioaccumulated toxic elements and their methods of excretion from the human body. This study was designed to assess the concentration of various toxic elements in three body fluids: blood, urine and sweat. Blood, urine, and sweat were collected from 20 individuals (10 healthy participants and 10 participants with various health problems) and analyzed for approximately 120 various compounds, including toxic elements. Toxic elements were found to differing degrees in each of blood, urine, and sweat. Serum levels for most metals and metalloids were comparable with those found in other studies in the scientific literature. Many toxic elements appeared to be preferentially excreted through sweat. Presumably stored in tissues, some toxic elements readily identified in the perspiration of some participants were not found in their serum. Induced sweating appears to be a potential method for elimination of many toxic elements from the human body. Biomonitoring for toxic elements through blood and/or urine testing may underestimate the total body burden of such toxicants. Sweat analysis should be considered as an additional method for monitoring bioaccumulation of toxic elements in humans.


An ultrasensitive electrochemical cytosensor for highly specific detection of HL-60 cancer cells based on metal ion functionalized titanium phosphate nanospheres.
Facile and sensitive detection methods of cancer cells in the early stage are beneficial for monitoring cancers and treating patients in time to reduce the death rate. In this work, an ultrasensitive cytosensor was constructed using aptamers as cell capturers and metal ion-exchanged titanium phosphate nanospheres as electrochemical probes. KH1C12 can specifically recognize HL-60 cells and distinguish them from other cell lines, K562 and CCRF-CEM, to obtain high selectivity. Cadmium ion functionalized titanium phosphate nanospheres show large quantities of electroactive cadmium ion output and a highly sensitive electrochemical signal. This proposed cytosensor showed a wide dynamic linear range from 102 cells per mL to 107 cells per mL with a low detection limit of 35 cells per mL, providing a new, simple and ultrasensitive platform for cancer diagnosis in biomedical and clinical research.


A quantitative trait locus influencing anxiety in the laboratory rat.
A critical test for a gene that influences susceptibility to fear in animals is that it should have a consistent pattern of effects across a broad range of conditioned and unconditioned models of anxiety. Despite many years of research, definitive evidence that genetic effects operate in this way is lacking. The limited behavioral test regimes so far used in genetic mapping experiments and the lack of suitable multivariate methodologies have made it impossible to determine whether the quantitative trait loci (QTL) detected to date specifically influence fear-related traits. Here we report the first multivariate analysis to explore the genetic architecture of rodent behavior in a battery of animal models of anxiety. We have mapped QTLs in an F2 intercross of two rat strains, the Roman high and low avoidance rats, that have been selectively bred for differential response to fear. Multivariate analyses show that one locus, on rat chromosome 5, influences behavior in different models of anxiety. The QTL influences two-way active avoidance, conditioned fear, elevated plus maze, and open field activity but not acoustic startle response or defecation in a novel environment. The direction of effects of the QTL alleles and a coincidence between the behavioral profiles of anxiolytic drug and genetic action are consistent with the QTL containing at least one gene with a pleiotropic action on fear responses. As the neural basis of fear is conserved across species, we suggest that the QTL may have relevance to trait anxiety in humans.


Long-term outcome of endoscopic resection for intramucosal esophageal squamous cell cancer: a secondary analysis of the Japan Esophageal Cohort study.
Prospectively collected long-term data of patients undergoing endoscopic resection for superficial esophageal squamous cell carcinoma (ESCC) are limited. The aim of this study was to determine the prospectively collected long-term outcomes of endoscopic resection for ESCC as a secondary analysis of the Japan Esophageal Cohort (JEC) study. Patients who underwent endoscopic resection of intramucosal ESCC at 16 institutions between September 2005 and May 2010 were enrolled in the JEC study. All patients underwent endoscopic examination with iodine staining at 3 and 6 months after resection, and every 6 months thereafter. We investigated clinical courses after endoscopic resection, survival rates, and cumulative incidence of metachronous ESCC. 330 patients (mean age 67.0 years) with 396 lesions (mean size 20.4 mm) were included in the analysis. Lesions were diagnosed as high-grade intraepithelial neoplasia in 17.4 % and as squamous cell carcinoma in 82.6 % (limited to epithelium in 28.4 %, to lamina propria in 55.4 %, and to muscularis mucosa in 16.2 %). En bloc resection was achieved in 291 (73.5 %). The median follow-up period was 49.4 months. Local recurrences occurred in 13 patients (3.9 %) and were treated by endoscopic procedures. Lymph node metastasis occurred in two patients (0.6 %) after endoscopic resection. The 5-year overall, disease-specific, and metastasis-free survival rates were 95.1 %, 99.1 %, and 94.6 %, respectively. The 5-year cumulative incidence rate of metachronous ESCC was 25.7 %. Our study demonstrated that endoscopic resection is an effective treatment for intramucosal ESCC, with favorable long-term outcomes.


Stevia Leaf to Stevia Sweetener: Exploring Its Science, Benefits, and Future Potential.
Steviol glycoside sweeteners are extracted and purified from the Stevia rebaudiana Bertoni plant, a member of the Asteraceae (Compositae) family that is native to South America, where it has been used for its sweet properties for hundreds of years. With continued increasing rates of obesity, diabetes, and other related comorbidities, in conjunction with global public policies calling for reductions in sugar intake as a means to help curb these issues, low- and no-calorie sweeteners (LNCSs, also known as high-potency sweeteners) such as stevia are gaining interest among consumers and food manufacturers. This appeal is related to stevia being plant-based, zero calorie and with a sweet taste that is 50-350 times sweeter than sugar, making it an excellent choice for use in sugar- and calorie-reduced food and beverage products. Despite the fact that the safety of stevia has been affirmed by several food regulatory and safety authorities around the world, insufficient education about stevia's safety and benefits, including continuing concern with regard to the safety of LNCSs in general, deters health professionals and consumers from recommending or using stevia. Therefore, the aim of this review and the stevia symposium that preceded this review at the ASN's annual conference in 2017 was to examine, in a comprehensive manner, the state of the science for stevia, its safety and potential health benefits, and future research and application. Topics covered included metabolism, safety and acceptable intake, dietary exposure, impact on blood glucose and insulin concentrations, energy intake and weight management, blood pressure, dental caries, naturality and processing, taste and sensory properties, regulatory status, consumer insights, and market trends. Data for stevia are limited in the case of energy intake and weight management as well as for the gut microbiome; therefore, the broader literature on LNCSs was reviewed at the symposium and therefore is also included in this review.


p53-dependent and independent expression of p21 during cell growth, differentiation, and DNA damage.
Expression of p21 has been shown to be up-regulated by the p53 tumor suppressor gene in vitro in response to DNA-damaging agents. However, p21 expression can be regulated independently of p53, and here we show that expression of p21 in various tissues during development and in the adult mouse occurs in the absence of p53 function. However, most tissues tested did require p53 for p21 induction following exposure of the whole animal to gamma irradiation. These results show that normal tissue expression of p21 to high levels is not dependent on p53 and confirm that induction of p21 by DNA-damaging agents does require p53. p21 is expressed upon differentiation of p53-deficient murine erythroleukemia (MEL) cells, and the kinetics of induction of p21 in this system suggest that it may be involved in the growth arrest that precedes terminal differentiation. The gene is up-regulated in mouse fibroblasts in response to serum restimulation but the kinetics and levels of induction differ between wild-type and mutant cells. Expression of p21 message following serum restimulation is superinducible by cycloheximide in wild-type but not in p53-deficient cells. The increases in p21 mRNA are reflected in changes in p21 protein levels. p21 expression also appears to be regulated at the post-transcriptional level because moderate increases in mRNA expression, during differentiation of MEL cells and upon serum restimulation of fibroblasts, are followed by large increases in protein levels. Regulation of the mouse p21 promoter by p53 depends on two critical p53-binding sites located 1.95 and 2.85 kb upstream from the transcriptional initiation site. The sequences mediating serum responsiveness of the promoter map to a region containing the proximal p53 site. p53 appears to play a critical role in p21 induction following DNA damage. Moreover, p21 can be regulated independently of p53 in several situations including during normal tissue development, following serum stimulation, and during cellular differentiation.


Centres for Leadership: a strategy for academic integration.
Purpose The purpose of this paper is to describe how an Academic Health Science Centre, providing pediatric rehabilitation services, research, and education, developed a Centres for Leadership (CfL) initiative to integrate its academic functions and embrace the goal of being a learning organization. Design/methodology/approach Historical documents, tracked output information, and staff members' insights were used to describe the ten-year evolution of the initiative, its benefits, and transformational learnings for the organization. Findings The evolutions concerned development of a series of CfLs, and changes over time in leadership and management structure, as well as in operations and targeted activities. Benefits included enhanced clinician engagement in research, practice-based research, and impacts on clinical practice. Transformational learnings concerned the importance of supporting stakeholder engagement, fostering a spirit of inquiry, and fostering leaderful practice. These learnings contributed to three related emergent outcomes reflecting "way stations" on the journey to enhanced evidence-informed decision making and clinical excellence: enhancements in authentic partnerships, greater innovation capacity, and greater understanding and actualization of leadership values. Practical implications Practical information is provided for other organizations interested in understanding how this initiative evolved, its tangible value, and its wider benefits for organizational collaboration, innovation, and leadership values. Challenges encountered and main messages for other organizations are also considered. Originality/value A strategy map is used to present the structures, processes, and outcomes arising from the initiative, with the goal of informing the operations of other organizations desiring to be learning organizations.


The treatment of restless legs syndrome and periodic limb movement disorder. An American Academy of Sleep Medicine Review.
A task force consisting of six authors reviewed the published literature on the therapy of the restless legs syndrome or periodic limb movements in sleep available in indices through April, 1998. They selected the 45 articles for detailed review which presented original investigations of therapeutic impact on the restless legs syndrome (RLS) or periodic limb movements (PLM) and which met minimal standards. These articles dealt with a range of pharmacological and other treatment modalities, although most dealt with medications and almost half of those concentrated on dopaminergic agents, especially levodopa in various formulations. Almost half of the articles reviewed used controlled methodologies, most commonly cross-over methodologies with randomized allocation of subjects. Multi-center studies with large numbers of subjects and long-term controlled studies were not found. Information was extracted from the articles and study design, clinical definition, evaluative measures, side effects, and outcomes were tabulated in 6 evidence tables and summarized in the accompanying text. This literature was evaluated for the nature of the studies performed and its coverage of potential therapies. The review concludes with comments on possible future directions for therapeutic investigation based on the current state of the literature.