Plasma fatty acid composition and incident ischemic stroke in middle-aged adults: the Atherosclerosis Risk in Communities (ARIC) Study.
The association of individual fatty acids with ischemic stroke has not been thoroughly studied, and results have been inconsistent. Few prospective studies have systematically explored the association of biomarkers of fatty acid intake with stroke. The aim of this study was to explore which individual plasma fatty acids would be associated with higher risk of ischemic stroke among whites. We studied 3,870 white men and women from the Minneapolis field center of the Atherosclerosis Risk in Communities (ARIC) Study, aged 45-64 years at baseline (1987-1989), who had plasma cholesterol ester (CE) and phospholipid (PL) fatty acids measured. Participants were followed through 2008 for incident ischemic stroke. Hazard ratios (HRs) with 95% confidence intervals (CIs) across quartiles of each fatty acid, measured as the percentage of total fatty acids, were calculated using the Cox proportional hazards model. During a maximum of 22 years of follow-up, we identified 168 cases of ischemic stroke. After adjustment for age and sex, plasma levels of saturated fatty acids were associated positively: HR (95% CI) of the highest versus the lowest quartile for CE fraction was 1.93 (1.23-3.04, p for trend = 0.01) and that for PL fraction was 1.64 (1.05-2.57, p for trend = 0.03). There was also a positive linear association with monounsaturated fatty acids, especially with palmitoleic acid: HR (95% CI) of the highest versus the lowest quartile for CE fraction was 1.86 (1.20-2.87, p for trend = 0.003) and that for PL fraction was 1.52 (0.99-2.34, p for trend = 0.005). No associations of ω-3 and ω-6 polyunsaturated fatty acids with ischemic stroke were observed, but linoleic acid was inversely and nonlinearly associated with ischemic stroke: HR (95% CI) of the highest versus the lowest quartile for CE fraction was 0.64 (0.43-0.97, p for trend = 0.13) and that for PL fraction was 0.69 (0.45-1.05, p for trend = 0.24). These associations were generally unchanged after adjustment for cardiovascular risk factors. In this US cohort of whites, we found significant positive associations of plasma saturated and monounsaturated fatty acids, especially of palmitoleic acid, with ischemic stroke. We also found an inverse nonlinear association between linoleic acid and ischemic stroke.


After the Battlefield: Infectious Complications among Wounded Warriors in the Trauma Infectious Disease Outcomes Study.
During recent wars in Iraq and Afghanistan, improved survivability in severe trauma corresponded with a rise in the proportion of trauma-related infections, including those associated with multidrug-resistant organisms (MDROs). Significant morbidity was reported in association with the infections. There is also concern regarding potential long-term impacts of the trauma-related infectious complications. Therefore, to meet the critical need of prospective collection of standardized infection-related data to understand the disease burden and improve outcomes of wounded personnel, the Trauma Infectious Disease Outcomes Study (TIDOS) was developed. Herein, we review accomplishments and key peer-reviewed findings of TIDOS. The TIDOS project is a multicenter observational study of short- and long-term infectious complications following deployment-related trauma. Wounded military personnel medevac'd to Landstuhl Regional Medical Center (LRMC; Germany) before transfer to a participating US military hospital between June 2009 and December 2014 were eligible for inclusion. An infectious disease module to supplement the Department of Defense Trauma Registry by collecting infection-related data from all trauma patients admitted to participating hospitals was developed. Specimens from trauma patients were also collected and retained in a microbiological isolate repository. During the initial hospitalization, patients were given the opportunity to enroll in a prospective follow-up cohort study. Patients who received Department of Veterans Affairs (VA) care were also given the opportunity to consent to ongoing VA follow-up. A total of 2,699 patients transferred to participating military hospitals in the USA, of which 1,359 (50%) patients enrolled in the TIDOS follow-up cohort. In addition, 638 enrolled in the TIDOS-VA cohort (52% of TIDOS enrollees who entered VA healthcare). More than 8,000 isolates were collected from infection control surveillance and diagnostic evaluations and retained in the TIDOS Microbiological Repository. Approximately 34% of the 2,699 patients at US hospitals developed a trauma-related infection during their initial hospitalization with skin and soft-tissue infections being predominant. After discharge from the US hospitals, approximately one-third of TIDOS cohort enrollees developed a new trauma-related infection during follow-up and extremity wound infections (skin and soft-tissue infections and osteomyelitis) continued to be the majority. Among TIDOS cohort enrollees who received VA healthcare, 38% developed a new trauma-related infection with the incident infection being diagnosed a median of 88 days (interquartile range: 19-351 days) following hospital discharge. Data from TIDOS have been used to support the development of Joint Trauma System clinical practice guidelines for the prevention of combat-related infections, as well as the management of invasive fungal wound infections. Lastly, due to the increasing proportion of infections associated with MDROs, TIDOS investigators have collaborated with investigators across military laboratories as part of the Multidrug-Resistant and Virulent Organisms Trauma Infections Initiative with the objective of improving the understanding of the complex wound microbiology in order to develop novel infectious disease countermeasures. The TIDOS project has focused research on four initiatives: (1) blast-related wound infection epidemiology and clinical management; (2) DoD-VA outcomes research; (3) Multidrug- Resistant and other Virulent Organisms Trauma Infections Initiative; and (4) Joint Trauma System clinical practice guidelines and antibiotic stewardship. There is a continuing need for longitudinal data platforms to support battlefield wound research and clinical practice guideline recommendation refinement, particularly to improve care for future conflicts. As such, maintaining a research platform, such as TIDOS, would negate the lengthy time needed to initiate data collection and analysis.


BEC, a novel enterotoxin of Clostridium perfringens found in human clinical isolates from acute gastroenteritis outbreaks.
Clostridium perfringens is a causative agent of food-borne gastroenteritis for which C. perfringens enterotoxin (CPE) has been considered an essential factor. Recently, we experienced two outbreaks of food-borne gastroenteritis in which non-CPE producers of C. perfringens were strongly suspected to be the cause. Here, we report a novel enterotoxin produced by C. perfringens isolates, BEC (binary enterotoxin of C. perfringens). Culture supernatants of the C. perfringens strains showed fluid-accumulating activity in rabbit ileal loop and suckling mouse assays. Purification of the enterotoxic substance in the supernatants and high-throughput sequencing of genomic DNA of the strains revealed BEC, composed of BECa and BECb. BECa and BECb displayed limited amino acid sequence similarity to other binary toxin family members, such as the C. perfringens iota toxin. The becAB genes were located on 54.5-kb pCP13-like plasmids. Recombinant BECb (rBECb) alone had fluid-accumulating activity in the suckling mouse assay. Although rBECa alone did not show enterotoxic activity, rBECa enhanced the enterotoxicity of rBECb when simultaneously administered in suckling mice. The entertoxicity of the mutant in which the becB gene was disrupted was dramatically decreased compared to that of the parental strain. rBECa showed an ADP-ribosylating activity on purified actin. Although we have not directly evaluated whether BECb delivers BECa into cells, rounding of Vero cells occurred only when cells were treated with both rBECa and rBECb. These results suggest that BEC is a novel enterotoxin of C. perfringens distinct from CPE, and that BEC-producing C. perfringens strains can be causative agents of acute gastroenteritis in humans. Additionally, the presence of becAB on nearly identical plasmids in distinct lineages of C. perfringens isolates suggests the involvement of horizontal gene transfer in the acquisition of the toxin genes.


Getting to a feasible income equality.
Income inequality is known to have negative impacts on an economic system, thus has been debated for a hundred years past or more. Numerous ideas have been proposed to quantify income inequality, and the Gini coefficient is a prevalent index. However, the concept of perfect equality in the Gini coefficient is rather idealistic and cannot provide realistic guidance on whether government interventions are needed to adjust income inequality. In this paper, we first propose the concept of a more realistic and 'feasible' income equality that maximizes total social welfare. Then we show that an optimal income distribution representing the feasible equality could be modeled using the sigmoid welfare function and the Boltzmann income distribution. Finally, we carry out an empirical analysis of four countries and demonstrate how optimal income distributions could be evaluated. Our results show that the feasible income equality could be used as a practical guideline for government policies and interventions.


Hypnotherapy for sleep disorders.
Hypnosis can be defined as a procedure during which changes in sensations, perceptions, thoughts, feelings or behaviour are suggested. Hypnosis can be used to amplify whatever it is about therapy that makes it therapeutic. It permits a wide range of choices regarding where and how to intervene in the patient's problems. In this paper, we set out to examine the rationale of using hypnotherapy to manage various types of sleep disorders, and to explore the techniques, strategies and hypnotic scripts employed by various hypnotherapists. We also examine the research data available on the efficacy of hypnosis in the treatment of sleep disorders. Acute and chronic insomnia often respond to relaxation and hypnotherapy approaches, along with sleep hygiene instructions. Hypnotherapy has also helped with nightmares and sleep terrors. There are several reports of successful use of hypnotherapy for parasomnias, specifically for head and body rocking, bedwetting and sleepwalking. Hypnosis is a specialised technique, not a therapy itself, and should be used as an adjunctive intervention within a complete psychological and medical treatment package. Most of the literature is limited to case reports or studies with such a small sample that at times it is very difficult to interpret the results. There is a major placebo effect, so uncontrolled trials are of limited value. It is hard to perform a randomised, double-blind, controlled trial to evaluate hypnotherapy given that cooperation and rapport between patient and therapist is needed to achieve a receptive trance state.


Factors contributing to medication noncompliance in elderly public housing tenants.
A study examined the extent and correlates of noncompliance in 155 predominantly elderly public housing tenants. Data were gathered through in-home interviews; only data from tenants taking prescribed medications on a regularly scheduled basis (n = 140) were analyzed. Noncompliance was significantly (p less than 0.05) associated with taking more than five prescribed medications, an inability to read prescription and auxiliary labels, and difficulty opening flip-off type medication container lids. The results indicate that attempts to reduce the total number of prescription drug products used in these elderly residents by carefully scrutinizing the need for each prescription drug may enhance compliance. Pharmacists could assist in the detection of elderly patients at risk for medication noncompliance (five or more total prescription medications) by assessing the patients' ability to read labels on prescription containers for all new drug additions and refills, and by having them try to open a flip-off container lid.


A Drosophila genetic screen yields allelic series of core microRNA biogenesis factors and reveals post-developmental roles for microRNAs.
Canonical animal microRNAs (miRNAs) are ∼22-nt regulatory RNAs generated by stepwise cleavage of primary hairpin transcripts by the Drosha and Dicer RNase III enzymes. We performed a genetic screen using an miRNA-repressed reporter in the Drosophila eye and recovered the first reported alleles of fly drosha, an allelic series of its dsRBD partner pasha, and novel alleles of dicer-1. Analysis of drosha mutants provided direct confirmation that mirtrons are independent of this nuclease, as inferred earlier from pasha knockouts. We further used these mutants to demonstrate in vivo cross-regulation of Drosha and Pasha in the intact animal, confirming remarkable conservation of a homeostatic mechanism that aligns their respective levels. Although the loss of core miRNA pathway components is universally lethal in animals, we unexpectedly recovered hypomorphic alleles that gave adult escapers with overtly normal development. However, the mutant photoreceptor neurons exhibited reduced synaptic transmission, without accompanying defects in neuronal development or maintenance. These findings indicate that synaptic function is especially sensitive to optimal miRNA pathway function. These allelic series of miRNA pathway mutants should find broad usage in studies of miRNA biogenesis and biology in the Drosophila system.


How do changes to the built environment influence walking behaviors? A longitudinal study within a university campus in Hong Kong.
Previous studies testing the association between the built environment and walking behavior have been largely cross-sectional and have yielded mixed results. This study reports on a natural experiment in which changes to the built environment were implemented at a university campus in Hong Kong. Longitudinal data on walking behaviors were collected using surveys, one before and one after changes to the built environment, to test the influence of changes to the built environment on walking behavior. Built environment data are from a university campus in Hong Kong, and include land use, campus bus services, pedestrian network, and population density data collected from campus maps, the university developmental office, and field surveys. Walking behavior data were collected at baseline in March 2012 (n = 198) and after changes to the built environment from the same cohort of subjects in December 2012 (n = 169) using a walking diary. Geographic information systems (GIS) was used to map walking routes and built environment variables, and compare each subject's walking behaviors and built environment exposure before and after the changes to the built environment. Walking behavior outcomes were changes in: i) walking distance, ii) destination-oriented walking, and iii) walked altitude range. Multivariable linear regression models were used to test for associations between changes to the built environment and changes in walking behaviors. Greater pedestrian network connectivity predicted longer walking distances and an increased likelihood of walking as a means of transportation. The increased use of recreational (vs. work) buildings, largely located at mid-range altitudes, as well as increased population density predicted greater walking distances.Having more bus services and a greater population density encouraged people to increase their walked altitude range. In this longitudinal study, changes to the built environment were associated with changes in walking behaviors. Use of GIS combined with walking diaries presents a practical method for mapping and measuring changes in the built environment and walking behaviors, respectively. Additional longitudinal studies can help clarify the relationships between the built environment and walking behaviors identified in this natural experiment.


[Impact assessment of increasing the time of withdrawal of colonoscopy in the detection rate of polyps in our midst].
In 2002, the US Multi-Society Task Force on Colorectal Cancer recommended that the withdrawal phase during colonoscopy should average at least 6-10 min to improve the detecting ratio of polyps. However, these recommendations were based on previous studies with a high colorectal cancer-risk population. The aim of our study is to determine if the implementation of a policy of colonoscopy withdrawal time > or = 6 minutes would be statistically associated with an increase in the colon polyp detection rate. We retrospectively evaluated the procedural data from all total colonoscopies performed at our general hospital from January 2006 through July 2009. Since January 2008 we implemented the policy of withdrawal time > or = 6 minutes. We calculated the detection rate and compared data from these period with our baseline consisted of the two years previous period in which withdrawal time was not aware. We reviewed procedural data from 2004 colonoscopies performed from January 2006 through July 2009; 1042 from the > or = 6 minutes-withdrawal time implementation period and 962 from the two years previous period. We found 162 patients with at least one polyp detected during the > or = 6 minutes-withdrawal time implementation period whereas 144 patients were detected from our baseline. The polyp detection rate was 15.55% from the period with a withdrawal time of e6 minutes versus 16.17 and 15.84% from each year of our baseline. No significant increase (p = 0.955) in the polyp detection rate was detected between the two comparative groups. The implementation of a policy of colonoscopy withdrawal time e6 minutes had no effect on colon polyp detection.


Assessing Correlation of Residency Applicants' Interview Dates With Likelihood of Matching.
This study aimed to determine whether the timing of an interview relative to the recruitment season was associated with being ranked or matched at an academic medical center. Eleven specialties (anesthesiology, diagnostic radiology, emergency medicine, family medicine, general surgery, internal medicine, neurology, neurosurgery, obstetrics-gynecology, orthopedic surgery, and psychiatry) that participated in the National Resident Matching Program were included in the study. Each program's total number of interview days during the October 2014-January 2015 interview season were divided equally into three interview time periods. The Cochran-Armitage trend test was used to evaluate associations among the three interview time periods (early, middle, and late) and interviewee outcomes (ranked or matched at our institution) for all subjects combined for each of the 11 programs and for specialty groups (medical, surgical, and hospital). Of 1034 applicants included in the analyses, 60% were men. Most were graduated from US medical schools (59.8%; a total of 103 applicants obtained first-year training positions through the Match [95.4% combined fill rate]). Twenty-nine interviewed early, 38 in the middle, and 36 in the late period (P = 0.3877). A total of 864 applicants were ranked by 1 of the 11 residency programs at the study site: 267 in the early period, 319 in the middle, and 278 in the late period (P = 0.4184). Being ranked in association with specialty classification also showed no significant differences. Interview timing had no relation to the likelihood of a match or being ranked by 1 of the 11 programs studied at our institution. These findings help dispel misconceptions about the importance of the interview date for a successful match.


Is testosterone a friend or a foe of the prostate?
Is there any unequivocal evidence that testosterone (T) can stimulate growth and aggravate symptoms in men with locally advanced and metastatic prostate cancer (PCa)? This is not a controversial point: the answer is yes. However, this evidence does not imply that PCa is a result of T or therapy with T (TTh) of hypogonadal men. Furthermore, currently adequately powered and optimally designed long-term prostate disease data are not available to determine if there is an additional risk from normal T values in cured patients for PCa. This Controversy is introduced by an endocrinologist, the section editor (E.A.J.) with G.L.G., a fellow urologist and radiotherapist expert in basic research on PCa. Two outstanding urologists, A.M and W.J.G.H., debate clinical data and clinical guidelines, respectively. Finally, other controversial issues are discussed by another leader in the field (A.M.) and a radiation oncologist and sexologist who is actually president of the International Society for Sexuality and Cancer (L.I.). Expert opinion supported by the critical review of the currently available literature. The answer to the main question "is the prostate a really T-dependent tissue?" is definitively yes, but T stimulates the prostatic tissue in a dose-dependent fashion only to a saturation point, achieved at low T concentrations. At these low T concentrations, stimulation is near maximal, and T supplementation above this level would not lead to significantly greater stimulation. Furthermore, there is no conclusive evidence that TTh increases the risk of PCa or even prostatic hyperplasia. There is also no evidence that TTh will convert subclinical PCa to clinically detectable PCa. However, there is a limited clinical experience of TTh after successful treatment of PCa. So far, just 48 patients have been studied in the three published articles. It is evident that the issue is still controversial and much more research is needed. However, the available data suggest to the expert in sexual medicine that TTh can be cautiously considered in selected hypogonadal men previously treated for curative intent of low-risk PCa and without evidence of active disease.


Fruit and Vegetable Intake and Risk of Posttransplantation Diabetes in Renal Transplant Recipients.
Posttransplantation diabetes mellitus (PTDM) contributes to risk for cardiovascular morbidity and mortality in renal transplant recipients (RTRs). In the general population, consumption of a diet containing few fruits and vegetables predisposes to type 2 diabetes. The role of diet as a potential modifiable risk factor for PTDM has not been explored. Our focus was to investigate the prospective associations of fruit and vegetable intake with risk of PTDM in stable RTRs. We included 472 adult RTRs who had a functioning graft ≥1 year. Fruit and vegetable intake was assessed by using a 177-item food frequency questionnaire. PTDM was defined according the American Diabetes Association's diagnostic criteria for diabetes. During 5.2 years of follow-up, 52 RTRs (11%) developed PTDM. Fruit intake was not associated with PTDM (hazard ratio [HR] 0.90 [95% CI 0.79-1.03] per 2log g/day; P = 0.13), whereas vegetable intake was inversely associated with PTDM (HR 0.77 [95% CI 0.63-0.94] per 2log g/day; P = 0.009). Mediation analyses revealed that ±50% of the association between vegetable intake and PTDM was mediated by variations in key components of the metabolic syndrome (i.e., HDL cholesterol, triglycerides, and waist circumference) as determined by the National Cholesterol Education Program's Adult Treatment Panel III Expert Panel. In this study vegetable intake, but not fruit intake, was associated with lower risk of PTDM in RTRs, likely largely through beneficial effects on key components of the metabolic syndrome. These findings further support accumulating evidence that supports a recommendation of higher vegetable intake by RTRs.


An active dimanganese(III)-tyrosyl radical cofactor in Escherichia coli class Ib ribonucleotide reductase.
Escherichia coli class Ib ribonucleotide reductase (RNR) converts nucleoside 5'-diphosphates to deoxynucleoside 5'-diphosphates and is expressed under iron-limited and oxidative stress conditions. This RNR is composed of two homodimeric subunits: alpha2 (NrdE), where nucleotide reduction occurs, and beta2 (NrdF), which contains an unidentified metallocofactor that initiates nucleotide reduction. nrdE and nrdF are found in an operon with nrdI, which encodes an unusual flavodoxin proposed to be involved in metallocofactor biosynthesis and/or maintenance. Ni affinity chromatography of a mixture of E. coli (His)(6)-NrdI and NrdF demonstrated tight association between these proteins. To explore the function of NrdI and identify the metallocofactor, apoNrdF was loaded with Mn(II) and incubated with fully reduced NrdI (NrdI(hq)) and O(2). Active RNR was rapidly produced with 0.25 +/- 0.03 tyrosyl radical (Y*) per beta2 and a specific activity of 600 units/mg. EPR and biochemical studies of the reconstituted cofactor suggest it is Mn(III)(2)-Y*, which we propose is generated by Mn(II)(2)-NrdF reacting with two equivalents of HO(2)(-), produced by reduction of O(2) by NrdF-bound NrdI(hq). In the absence of NrdI(hq), with a variety of oxidants, no active RNR was generated. By contrast, a similar experiment with apoNrdF loaded with Fe(II) and incubated with O(2) in the presence or absence of NrdI(hq) gave 0.2 and 0.7 Y*/beta2 with specific activities of 80 and 300 units/mg, respectively. Thus NrdI(hq) hinders Fe(III)(2)-Y* cofactor assembly in vitro. We propose that NrdI is an essential player in E. coli class Ib RNR cluster assembly and that the Mn(III)(2)-Y* cofactor, not the diferric-Y* one, is the active metallocofactor in vivo.


Projections of Atmospheric Nitrogen Deposition to the Chesapeake Bay Watershed.
Atmospheric deposition is among the largest pathways of nitrogen loading to the Chesapeake Bay Watershed (CBW). The interplay between future climate and emission changes in and around the CBW will likely shift the future nutrient deposition abundance and chemical regime (e.g., oxidized vs. reduced nitrogen). In this work, a Representative Concentration Pathway (RCP) from the Community Earth System Model is dynamically downscaled using the Weather Research and Forecasting (WRF) and Community Multiscale Air Quality (CMAQ) model coupled to the agro-economic Environmental Policy Integrated Climate (EPIC) model. The relative impacts of emission and climate changes on atmospheric nutrient deposition are explored for a recent historical period and a period centered on 2050. The projected regional emissions in CMAQ reflect current federal and state regulations, which use baseline and projected emission years 2011 and 2040, respectively. The historical simulations of 2-m temperature and precipitation have cool and dry biases, and temperature and precipitation are projected to both increase. Ammonium wet deposition agrees well with observations, but nitrate wet deposition is underpredicted. Climate and deposition changes increase simulated future ammonium fertilizer application. In the CBW at 2050, these changes (along with widespread decreases in anthropogenic nitrogen oxide and sulfur oxide emissions, and relatively constant NH3 emissions) decrease total nitrogen deposition by 21%, decrease annual average oxidized nitrogen deposition by 44%, and increase reduced nitrogen deposition by 10%. These results emphasize the importance of decreased anthropogenic emissions on the control of future nitrogen loading to the Chesapeake Bay in a changing climate.


Acoustially-mediated microfluidic nanofiltration through graphene films.
We exploit the possibility of enhancing the molecular transport of liquids through graphene films using amplitude modulated surface acoustic waves (SAWs) to demonstrate effective and efficient nanoparticle filtration. The use of the SAW, which is an extremely efficient means for driving microfluidic transport, overcomes the need for the large mechanical pumps required to circumvent the large pressure drops encountered in conventional membranes for nanoparticle filtration. 100% filtration efficiency was obtained for micron-dimension particulates, decreasing to only 95% for the filtration of particles of tens of nanometers in dimension, which is comparable to that achieved with other methods. To circumvent clogging of the film, which is typical with all membrane filters, a backwash operation to flush the nanoparticles is incorporated simply by reversing the SAW-induced flow such that 98% recovery of the initial filtration rate is recovered. Given these efficiencies, together with the low cost and compact size of the chipscale SAW devices, we envisage the possibility of scaling out the process by operating a large number of devices in parallel to achieve typical industrial-scale throughputs with potential benefits in terms of substantially lower capital, operating and maintenance costs.


Ambient odors of orange and lavender reduce anxiety and improve mood in a dental office.
The goal of this study was to investigate the impact of the essential oils of orange and lavender on anxiety, mood, alertness and calmness in dental patients. Two hundred patients between the ages of 18 and 77 years (half women, half men) were assigned to one of four independent groups. While waiting for dental procedures patients were either stimulated with ambient odor of orange or ambient odor of lavender. These conditions were compared to a music condition and a control condition (no odor, no music). Anxiety, mood, alertness and calmness were assessed while patients waited for dental treatment. Statistical analyses revealed that compared to control condition both ambient odors of orange and lavender reduced anxiety and improved mood in patients waiting for dental treatment. These findings support the previous opinion that odors are capable of altering emotional states and may indicate that the use of odors is helpful in reducing anxiety in dental patients.


A phase I study of intravenous TZT-1027 administered on day 1 and day 8 of a three-weekly cycle in combination with carboplatin given on day 1 alone in patients with advanced solid tumours.
TZT-1027 is a tubulin-binding drug and synthetic derivative of dolastatin-10 with cytotoxic and antivascular activity in vitro and in vivo. Studies have demonstrated anti-tumour activity in several tumour types. Patients were treated with escalating doses of TZT-1027 and carboplatin at doses from 1.6 to 2.0 mg/m2 and AUC 4 and 5 respectively. For pharmacokinetic analysis, plasma sampling was done during the first course using a high-performance liquid chromatographic assay. 14 patients received a total of 55 cycles at three dose levels. Dose limiting toxicities (DLTs) were first observed with 1.6 mg/m2 TZT-1027 and carboplatin AUC 5; 1 patient had grade 4 neutropenia and a delay in day 8 treatment occurred in two patients (gr 2 fatigue, gr 3 diarrhoea). At TZT-1027 2 mg/m2 and carboplatin AUC 5, one patient experienced grade 3 paralytic ileus. The most frequent toxicities were neutropenia, anaemia, fatigue, constipation, infection and vomiting. Peripheral neuropathy was reported in 36% of patients. One patient (pancreatic adenocarcinoma) achieved a partial response lasting 181 days. Pharmacokinetic analysis did not demonstrate any interaction between TZT-1027 and carboplatin. The recommended phase II dose is TZT-1027 1.6 mg/m2 and carboplatin AUC 5. No evidence of a PK interaction between these agents was observed.


Blood parameters in fattening pigs fed whole-ear corn silage and housed in group pens or in metabolic cages.
The aim of this work was to evaluate the effects of the inclusion of whole-ear corn silage (WECS) in diets for advanced fattening heavy pigs (substitution for part of the dry corn and wheat bran) allocated or not in metabolic cages on the main blood parameters. The high-moisture shelled corn is largely used in pig feeding while WECS is less often used despite the fact that it increases the DM crop yield. Three experimental diets were fed to 27 barrows (Italian Large White × Italian Duroc), with an average BW of 98.2 (±5.6) kg at the start of the trial, and randomly allotted to 3 experimental groups including a control diet (CON) containing cereal meals (corn, barley, and wheat, 80.2% DM in total), soybean meal (9% DM), wheat bran (8% DM), minerals and supplements (2.8% DM), and 2 diets containing WECS (15 or 30% DM referred to as 15WECS and 30WECS, respectively) in partial or complete substitution for wheat bran and corn meal. The pigs were randomly housed in 9 pens with 3 animals per pen and 3 pens per dietary treatment. Six pigs per each of the 3 treatments were moved from the pens to individual metabolic cages for 3 consecutive periods (2 pigs per treatment per period). Each period lasted 14 d, and blood was collected at the start and at the end of the periods. Blood was drawn from the jugular vein before feed distribution in the morning, at 14 d intervals, and analyzed for hematological, metabolic, and serum protein profiles. The effect of the metabolic cage housing was included in the statistical model to compare the results obtained in the 2 different environments of restrained and group-housed barrows. The WECS affected the neutrophil to lymphocyte ratio and mean corpuscular hemoglobin concentration. The main diet effect on plasma metabolites was recorded for plasma NEFA, with higher values in WECS diets compared with the CON. The metabolic cage housing affected both hematological (red blood cell count, hemoglobin, hematocrit) and metabolic (protein and its fractions) items, which can be markers of hemodilution. These results indicate the possibility to use this feed in the diet of heavy pigs without negative effects on physiology. The absolute values from metabolic profile of pigs in metabolic cages must be considered with caution for possible comparisons with values obtained on-field in group pens, particularly because a different hemodilution may affect the results.


Bayesian estimation of vaccine efficacy.
Protective vaccine efficacy, VEs, is measured as one minus the incidence rate ratio (IRR) or the relative risk (RR) in the vaccinated group compared with the unvaccinated group. In this paper, we systematically present Bayesian estimation of protective vaccine efficacy based on the Poisson and binomial distributions. We also propose a new tool, the vaccine efficacy acceptability curve, to represent the uncertainty for the estimate of the vaccine efficacy graphically. It is very useful, especially when there is no universal agreement on the acceptable vaccine efficacy. The vaccine efficacy acceptability curve is defined as the posterior probability that the measure of vaccine efficacy VEs > or = k for each acceptable value k. When a vaccine is highly efficacious, the number of vaccinated susceptibles being infected is likely to be very small or even zero. Then the assumptions of normality and log-normality of IRR or RR usually do not hold well. Although frequentist exact methods provide good estimates of the confidence interval, they are overly conservative and are computationally difficult to extend to estimate the vaccine efficacy acceptability curve. In this paper, our focus is on Bayesian estimation of protective vaccine efficacy, its highest probability density credible set, and the vaccine efficacy acceptability curve through Markov chain Monte Carlo (MCMC) methods. We illustrate the methods using the data from two pertussis vaccine studies and the H. influenza Type B preventive trial.


Thrombospondin-1 and its CSVTCG-specific receptor in wound healing and cancer.
Growth factors play a crucial role in the regulation of cellular proliferation and matrix degradation in wound healing and cancer. We have shown that thrombospondin 1 (TSP-1) and its cysteine-serine-valine-threonine-cysteine-glycine (CSVTCG)-specific receptor play a key role in cell invasion and matrix degradation in different carcinomas. The present study was done to determine whether TSP-1 and its receptor show a similar pattern of expression in wound healing and cancer. Expression and localization of TSP-1 and its receptor were determined in fetal wounds, adult burn wounds, and different human malignancies by immunohistochemical staining and computerized image analysis. In healing wounds, TSP-1 was expressed in the stroma early in the process, followed by a steep decline. The TSP-1 receptor localized to neovessels and highly proliferating cells (i.e., fibroblasts, basal cells), its levels remaining relatively constant. Cancer cells and tumor-associated microvessels expressed the TSP-1 receptor, whereas TSP-1 localized predominantly to the tumor-associated stroma. These data suggest a critical role for TSP-1 and its CSVTCG-specific receptor in wound healing and cancer.


An ethical template for pharmacy benefits.
We propose an ethical template for pharmacy benefits and a fair process for using it. The template delineates four levels of decisions about pharmacy coverage, connecting ethically acceptable types of rationales for limits with decisions made at each level. It provides a framework for organizing ethically relevant reasons for coverage (or the tiered copayments). The process for using the template assures accountability for the reasonableness of benefit decisions. It requires transparency and relevance of rationales for limit setting and revisability of decisions, including through fair procedures for appeals. The template and the process facilitate broader public learning about fair limit setting.


The role of the polycomb complex in silencing alpha-globin gene expression in nonerythroid cells.
Although much is known about globin gene activation in erythroid cells, relatively little is known about how these genes are silenced in nonerythroid tissues. Here we show that the human alpha- and beta-globin genes are silenced by fundamentally different mechanisms. The alpha-genes, which are surrounded by widely expressed genes in a gene dense region of the genome, are silenced very early in development via recruitment of the Polycomb (PcG) complex. By contrast, the beta-globin genes, which lie in a relatively gene-poor chromosomal region, are not bound by this complex in nonerythroid cells. The PcG complex seems to be recruited to the alpha-cluster by sequences within the CpG islands associated with their promoters; the beta-globin promoters do not lie within such islands. Chromatin associated with the alpha-globin cluster is modified by histone methylation (H3K27me3), and silencing in vivo is mediated by the localized activity of histone deacetylases (HDACs). The repressive (PcG/HDAC) machinery is removed as hematopoietic progenitors differentiate to form erythroid cells. The alpha- and beta-globin genes thus illustrate important, contrasting mechanisms by which cell-specific hematopoietic genes (and tissue-specific genes in general) may be silenced.


A survey of the Czechoslovak follow-up of lung cancer mortality in uranium miners.
The major Czechoslovak cohort of uranium miners (S-cohort) is surveyed in terms of diagrams illustrating dependences on calendar year, age, and exposure to radon and radon progeny. An analysis of the dose dependence of lung cancer mortality is performed by nonparametric and, subsequently, by parametric methods. In the first step, two-dimensional isotonic regression is employed to derive the lung cancer mortality rate and the relative excess risk as functions of age attained and of lagged cumulated exposure. In a second step, analytical fits in terms of relative risk models are derived. The treatment is largely analogous to the methods applied by the BEIR IV Committee to other major cohorts of uranium miners. There is a marked dependence of the excess risk on age attained and on time since exposure. A specific characteristic of the Czechoslovak data is the nonlinearity of the dependence of the lung cancer excess risk on the cumulated exposure; exposures on the order of 100 working level months or less appear to be more effective per working level month than larger exposures but, in the absence of an internal control group, this cannot be excluded to be due to confounders such as smoking or environmental exposures. A further notable observation is the association of larger excess risks with longer protraction of the exposures.


Can urinary PCA3 supplement PSA in the early detection of prostate cancer?
Given the limited sensitivity and specificity of prostate-specific antigen (PSA), its widespread use as a screening tool has raised concerns for the overdiagnosis of low-risk and the underdiagnosis of high-grade prostate cancer. To improve early-detection biopsy decisions, the National Cancer Institute conducted a prospective validation trial to assess the diagnostic performance of the prostate cancer antigen 3 (PCA3) urinary assay for the detection of prostate cancer among men screened with PSA. In all, 859 men (mean age, 62 years) from 11 centers scheduled for a diagnostic prostate biopsy between December 2009 and June 2011 were enrolled. The primary outcomes were to assess whether PCA3 could improve the positive predictive value (PPV) for an initial biopsy (at a score > 60) and the negative predictive value (NPV) for a repeat biopsy (at a score < 20). For the detection of any cancer, PPV was 80% (95% CI, 72% to 86%) in the initial biopsy group, and NPV was 88% (95% CI, 81% to 93%) in the repeat biopsy group. The addition of PCA3 to individual risk estimation models (which included age, race/ethnicity, prior biopsy, PSA, and digital rectal examination) improved the stratification of cancer and of high-grade cancer. These data independently support the role of PCA3 in reducing the burden of prostate biopsies among men undergoing a repeat prostate biopsy. For biopsy-naive patients, a high PCA3 score (> 60) significantly increases the probability that an initial prostate biopsy will identify cancer.


An evaluation of the potential factors affecting lifetime reproductive success in salmonids.
Lifetime reproductive success (LRS), the number of offspring produced over an organism's lifetime, is a fundamental component of Darwinian fitness. For taxa such as salmonids with multiple species of conservation concern, understanding the factors affecting LRS is critical for the development and implementation of successful conservation management practices. Here, we reviewed the published literature to synthesize factors affecting LRS in salmonids including significant effects of hatchery rearing, life history, and phenotypic variation, and behavioral and spawning interactions. Additionally, we found that LRS is affected by competitive behavior on the spawning grounds, genetic compatibility, local adaptation, and hybridization. Our review of existing literature revealed limitations of LRS studies, and we emphasize the following areas that warrant further attention in future research: (1) expanding the range of studies assessing LRS across different life-history strategies, specifically accounting for distinct reproductive and migratory phenotypes; (2) broadening the variety of species represented in salmonid fitness studies; (3) constructing multigenerational pedigrees to track long-term fitness effects; (4) conducting LRS studies that investigate the effects of aquatic stressors, such as anthropogenic effects, pathogens, environmental factors in both freshwater and marine environments, and assessing overall body condition, and (5) utilizing appropriate statistical approaches to determine the factors that explain the greatest variation in fitness and providing information regarding biological significance, power limitations, and potential sources of error in salmonid parentage studies. Overall, this review emphasizes that studies of LRS have profoundly advanced scientific understanding of salmonid fitness, but substantial challenges need to be overcome to assist with long-term recovery of these keystone species in aquatic ecosystems.


Oregon hospice nurses and social workers' assessment of physician progress in palliative care over the past 5 years.
The 1997 enactment of the Oregon Death with Dignity Act intensified interest in improving physician education and skills in caring for patients at the end of life. To obtain hospice nurse and social workers' collateral ratings of efforts made by Oregon physicians to improve their palliative care skills over the previous 5 years. A descriptive survey of nurses and social workers from all 50 Oregon outpatient hospice agencies. Oregon hospice nurse (N=185) and social worker (N=52) respondents, who had worked in hospice for at least 5 years, rated changes they observed over the past 5 years in physicians' approach to caring for their hospice clients. Six characteristics, including willingness to refer patients to hospice, willingness to prescribe sufficient pain medications, knowledge about using pain medications in hospice patients, interest in caring for hospice patients, competence in caring for hospice patients, and fearfulness of prescribing sufficient opioid medications were evaluated. Positive changes were endorsed by the majority of respondents on all but the scale measuring fearfulness of prescribing opioid medications; on the latter, 47% of nurses rated doctors as less fearful, whereas 53% rated them as about the same or more fearful than they were 5 years earlier. Most respondents rated Oregon physicians as showing improvements in knowledge and willingness to refer and care for hospice patients.


False-positive Cultures After Native Knee Aspiration: True or False.
Synovial fluid aspiration is a routine practice used by most orthopaedic surgeons to aid in the diagnosis of joint infection. In patients for whom there is a low pretest probability of infection, a positive culture-particularly if it is a broth-only culture-may be considered a contaminant, especially if the bacterial species are skin pathogens. To our knowledge no study has evaluated the incidence of contamination of aspirations from the native knee. What is the frequency of false-positive cultures among knee aspirations of the native knee? Two hundred patients, with a total of 200 knees, with the diagnosis of degenerative osteoarthritis undergoing a total knee arthroplasty (TKA) were identified for this study. None of these patients had symptoms, signs, or laboratory studies to suggest the presence of joint infection; a positive culture in this population therefore would be considered contaminated. Thirty-two (16%) patients were excluded secondary to a dry aspiration. One patient was enrolled in the study but did not have the knee aspirated and another patient's specimen was accidentally discarded. Each knee was aspirated under sterile conditions before performing the TKA. The fluid was sent for cell count and culture. If insufficient fluid was obtained for both cell count and culture, culture was performed rather than cell count. There were no false-positive cultures (zero of 166 [0%]) in aspirations of native knees. Our study would indicate that when done properly under sterile technique, cultures taken from knee arthrocentesis in patients without prosthetic joints should not be affected by perceived contaminant species. A positive specimen finding on culture should raise a strong suspicion of bacterial septic arthritis. Future studies should include more specimens as well as knees with prior TKA to help further identify the rate of false-positive cultures in knee arthrocentesis in both populations. Level I, diagnostic study.


Zika Virus and Patient Blood Management.
Sporadic Zika virus infections had only occurred in Africa and Asia until an outbreak in Micronesia (Oceania) in 2007. In 2013 to 2014, several outer Pacific Islands reported local outbreaks. Soon thereafter, the virus was likely introduced in Brazil from competing athletes from French Polynesia and other countries that participated in a competition there. Transmission is thought to have occurred through mosquito bites and spread to the immunologically naive population. Being also a flavivirus, the Zika virus is transmitted by the Aedes mosquito that is endemic in South and Central America that is also the vector of West Nile virus, dengue, and chikungunya. In less than a year, physicians in Brazil reported a many-fold increase in the number of babies born with microcephaly. Despite initial skepticism regarding the causal association of the Zika virus epidemic and birth defects, extensive basic and clinical research evidence has now confirmed this relationship. In the United States, more than 4000 travel-associated infections have been reported by the middle of 2016 to the Centers for Disease Control and Prevention. Furthermore, many local mosquito-borne infections have occurred in Puerto Rico and Florida. Considering that the virus causes a viremia in which 80% of infected individuals have no symptoms, the potential for transfusion transmission from an asymptomatic blood donor is high if utilizing donor screening alone without testing. Platelet units have been shown to infect 2 patients via transfusion in Brazil. Although there was an investigational nucleic acid test available for testing donors, not all blood centers were initially required to participate. Subsequently, the US Food and Drug Administration issued a guidance in August 2016 that recommended universal nucleic acid testing for the Zika virus on blood donors.In this report, we review the potentially devastating effects of Zika virus infection during pregnancy and its implication in cases of Guillain-Barre syndrome in adults. Furthermore, we urge hospital-based clinicians and transfusion medicine specialists to implement perisurgical patient blood management strategies to avoid blood component transfusions with their potential risks of emerging pathogens, illustrated here by the Zika virus. Ultimately, this current global threat, as described by the World Health Organization, will inevitably be followed by future outbreaks of other bloodborne pathogens; the principles and practices of perioperative patient blood management will reduce the risks from not only known, but also unknown risks of blood transfusion for our patients.


Anticancer effect of triterpenes from Ganoderma lucidum in human prostate cancer cells.
Ganoderma lucidum, within the Polyporaceae family of Basidiomycota, is a popular traditional remedy medicine used in Asia to promote health and longevity. Compounds extracted from G. lucidum have revealed anticancer, antioxidant and liver protective effects. G. lucidum has been associated with prostate cancer cells. G. lucidum extracts contain numerous bioactive components; however, the exact functional monomer is unknown and the role of triterpenes from G. lucidum (GLT) in prostate cancer remain obscure. The present study investigated the effects of GLT on cell viability, migration, invasion and apoptosis in DU-145 human prostate cancer cells. The results demonstrated that a high dose (2 mg/ml) of GLT inhibits cell viability in a dose- and time-dependent manner by the regulation of matrix metalloproteases. Furthermore, GLT induced apoptosis of DU-145 cells. In general, GLT exerts its effect on cancer cells via numerous mechanisms and may have potential therapeutic use for the prevention and treatment of cancer.


Is there a need for a common framework of dental specialties in Europe?
This paper aims to promote discussion about dental specialties and post-graduate dental education in the European Union (EU). Previously, dental educators have concentrated their efforts of seeking Pan-EU convergence in undergraduate dental education. However, the impact of the enlargement of the EU, the new European Commission (EC) Directive of professional training and the Bologna Process all impact on post-graduate (specialist) just as much as on undergraduate dental education. The provisions of the new EC directive mean that, unlike new medical specialties, new Pan-EU dental specialties cannot be created purely because they exist in two-fifths of EU Member States. At present, some EU Member States recognise eight or more dental specialties, whereas others recognise none. It is suggested that changing needs and demands of patients, which reflect a general improvement in oral health, increased wealth and an aging population will place increasing demands on dentistry to provide more complex care and treatment and that the current undergraduate curriculum cannot be expanded to provide suitable training to meet these needs and demands. There is thus a need to expand dental specialist training in all EU Member States, to agree common standards for specialist education and to officially recognise a wider range of Pan-EU dental specialties. The paper concludes that in order to achieve these goals, there is a need of a better collaboration between competent authorities, including governments, universities, dental associations and the various Pan-European Scientific Specialist Organizations.


Unexpected coordination number and phase diagram of niobium diselenide under compression.
We discovered several new energetically competitive structures of NbSe2 using the multi-algorithm collaborative (MAC) crystal structure prediction algorithm combined with the density functional theory. It was found that the coordination number of Nb in NbSe2 is increased from 6 to 7, and then to 8 with increasing pressure. Furthermore, it was unexpected that an Se atom would be squeezed to the center of a cage formed by 12 other Se atoms and then have 12-fold coordination when the pressure was increased to 130.4 GPa. The 12-coordination metalloid atom has never been discovered in other transition metal dichalcogenides. The new C2/m, I4/mmm, and P4/mmm NbSe2 were verified to be stable under both dynamically and mechanically stabile conditions. It is especially noteworthy that the new C2/m-NbSe2 was predicted to be potentially synthesized at high pressure and recovered under ambient conditions. A detailed high-pressure and high-temperature phase diagram was constructed based on the quasi-harmonic approximation up to 200 GPa, and the synthesis conditions of different new NbSe2 materials were also analyzed. All the discoveries in this study will guide the future synthesis of new NbSe2 materials at specific pressure and under temperature conditions and also help to further understand other transition metal dichalcogenides.


Written in Blood: Applying Shape Grammars to Retinal Vasculatures.
Blood vessel networks within the retina are crucial for maintaining tissue perfusion and therefore good vision. Their complexity and unique patterns often require a steep learning curve for humans to identify trends and changes in the shape and topology of the networks, even though there exists much information important to identifying disease within them. Through image processing, the vasculature is isolated from other features of the fundus images, forcing the viewer to focus on the complex vascular feature. This article explores an approach using a grammar based on shape to describe retinal vasculature and to generate realistic and increasingly unrealistic artificial vascular networks that are then reviewed by ophthalmologists via digital survey. The ophthalmologists are asked whether these artificial vascular networks appeared realistic or unrealistic. With only three rules (initiate, branch, and curve), the grammar accomplishes these goals. Networks are generated by adding noise to rule parameters present in existing networks. Via the survey of synthetic networks generated with different noise parameters, a correlation between noise in the branch rule and realistic association is revealed. By creating a language to describe retinal vasculature, this article allows for the potential of new insight into such an important but less understood feature of the retina, which in the future may play a role in diagnosing or helping to predict types of ocular disease. Applying shape grammar to describe retinal vasculature permits new understanding, which in turn provides the potential for new diagnostic tools.


Regression dilution methods for meta-analysis: assessing long-term variability in plasma fibrinogen among 27,247 adults in 15 prospective studies.
Within-person variability in measured values of a risk factor can bias its association with disease. The extent of this regression dilution bias for plasma fibrinogen was investigated using repeat measurement data collected at varying time intervals on 27 247 adults in 15 prospective studies. Regression dilution ratios (RDRs) were estimated from a linear regression of repeat measurements on baseline values in each study and for each time interval, and pooled allowing for within- and between-study heterogeneity. RDRs were estimated both without and with adjustment for confounders, and factors were investigated that might influence the RDRs. The unadjusted overall RDR was 0.51 (95% CI: 0.47, 0.55), which decreased to 0.46 (95% CI: 0.42, 0.49) after adjustment for age, sex and measured values of other established vascular risk factors. The RDR did not vary materially by assay method, age, sex or smoking status, but decreased at higher levels of baseline fibrinogen. It is appropriate to use an RDR of 0.5 to correct approximately for regression dilution bias in plasma fibrinogen values; however, this correction factor may produce somewhat conservative hazard ratios in adjusted analyses, at higher fibrinogen concentrations and in follow-up beyond a decade. More generally, the methods described in this report have widespread applicability to quantifying regression dilution bias in repeatability data from multiple prospective studies.


Real-time ray tracing of implicit surfaces on the GPU.
Compact representation of geometry using a suitable procedural or mathematical model and a ray-tracing mode of rendering fit the programmable graphics processor units (GPUs) well. Several such representations including parametric and subdivision surfaces have been explored in recent research. The important and widely applicable category of the general implicit surface has received less attention. In this paper, we present a ray-tracing procedure to render general implicit surfaces efficiently on the GPU. Though only the fourth or lower order surfaces can be rendered using analytical roots, our adaptive marching points algorithm can ray trace arbitrary implicit surfaces without multiple roots, by sampling the ray at selected points till a root is found. Adapting the sampling step size based on a proximity measure and a horizon measure delivers high speed. The sign test can handle any surface without multiple roots. The Taylor test that uses ideas from interval analysis can ray trace many surfaces with complex roots. Overall, a simple algorithm that fits the SIMD architecture of the GPU results in high performance. We demonstrate the ray tracing of algebraic surfaces up to order 50 and nonalgebraic surfaces including a Blinn's blobby with 75 spheres at better than interactive frame rates.


Interaction of halichondrin B and homohalichondrin B with bovine brain tubulin.
Halichondrin B is a polyether macrolide of marine origin which binds to tubulin and inhibits microtubule assembly in vitro and in vivo. As is the case with phomopsin A and dolastatin 10, halichondrin B is a non-competitive inhibitor of vinblastine binding to tubulin. Analogous to maytansine, which by contrast is a competitive inhibitor of vinblastine binding, halichondrin B has no effect on colchicine binding, which is greatly stabilized by phomopsin A and dolastatin 10, but not by maytansine. We have previously developed assays which allow sensitive discrimination among the interactions of various ligands with tubulin, and examined the effects of ligands on the reactivity of tubulin sulfhydryl groups and the exposure of hydrophobic areas on the surface of the tubulin molecule. To classify the nature of the interaction between halichondrin B and tubulin, in this study we examined the effects of halichondrin B and its closely related analogue, homohalichondrin B, by these assays. We found that: (1) halichondrin B and homohalichondrin B both inhibited formation of an intra-chain cross-link between two sulfhydryl groups in beta-tubulin, as do phomopsin A, dolastatin 10, maytansine, and vinblastine; (2) halichondrin B resembles maytansine in that it had no effect on alkylation of tubulin sulfhydryl groups by iodoacetamide, unlike phomopsin A, dolastatin 10 and vinblastine, all of which inhibit alkylation; (3) halichondrin B differs from other anti-mitotic drugs in that it enhanced exposure of hydrophobic areas on tubulin; (4) homohalichondrin B, like maytansine and in contrast to phomopsin A, dolastatin 10 and vinblastine, had no effect on exposure of hydrophobic areas; and (5) homohalichondrin B, contrary to maytansine, inhibited alkylation of tubulin sulfhydryl groups in the presence of GTP and MgCl2. In their interactions with the tubulin molecule, halichondrin B and homohalichondrin B appear to have unique conformational effects which differ from those of other drugs and also from the effects of each other as well.


Ionic mechanisms underlying the effects of vasoactive intestinal polypeptide on canine atrial myocardium.
Vasoactive intestinal polypeptide (VIP) is released from intracardiac neurons during vagal stimulation, ischemia, and heart failure, which are associated with increased vulnerability to atrial fibrillation. VIP shortens atrial effective refractory periods in dogs. Endogenous VIP contributes to vagally mediated acceleration of atrial electric remodeling. VIP is also shown to prolong the duration of acetylcholine-induced atrial fibrillation. However, the ionic mechanisms underlying VIP effects are largely unknown. The effects of VIP on transmembrane ion channels were studied in canine atrial cardiomyocytes using patch-clamp techniques. VIP increased delayed rectifier K+ current and L-type calcium current but decreased the transient outward K+ current and sodium current. Optical mapping technique was used to assess effects of VIP on action potential durations (APDs) in isolated canine left atria. VIP shortened APD and slowed conduction velocity in a dose-dependent manner. Furthermore, VIP increased spatial heterogeneity of APD and conduction velocity, as assessed by the SDs of APD and conduction velocity, and atrial fibrillation inducibility. Through its diverse effects on ion channels, VIP shortens APD with increased APD spatial heterogeneity and decreases intra-atrial conduction velocity, which may play an important role in the pathogenesis of atrial arrhythmias in scenarios where VIP release is increased.


Back to the Consideration of Future Consequences Scale: time to reconsider?
The Consideration of Future Consequences (CFC) Scale is a measure of the extent to which individuals consider and are influenced by the distant outcomes of current behavior. In this study, the authors conducted factor analysis to investigate the factor structure of the 12-item CFC Scale. The authors found evidence for a multiple factor solution including one completely present-oriented factor consisting of all 7 present-oriented items, and one or two future-oriented factors consisting of the remaining future-oriented items. Further evidence indicated that the present-oriented factor and the 12-item CFC Scale perform similarly in terms of internal consistency and convergent validity. The structure and content of the future-oriented factor(s) is unclear. From the findings, the authors raise questions regarding the construct validity of the CFC Scale, the interpretation of its results, and the usefulness of the CFC scale in its current form in applied research.


Evaluation of diagnostic accuracy of insulin-like growth factor (IGF)-I and IGF-binding protein-3 in growth hormone-deficient children and adults using ROC plot analysis.
We critically evaluated the diagnostic value of IGF-I and IGF-binding protein-3 (IGFBP-3) in GH deficiency (GHD) in children and adults using receiver operating characteristic (ROC) plot analysis. Sixty-six children (chronological age, 1.3-15 yr) were studied: 34 GHD and 32 idiopathic short stature (ISS). Ninety-two adults (chronological age, 18-70 yr) were also evaluated: 72 GHD, 34 of childhood onset (AGHD-CO), and 38 of adult onset (AGHD-AO); and 20 healthy volunteers. The SD score (SDS) for IGF-I was calculated from 596 normal subjects (212 children and 384 adults), and the SDS for IGFBP-3 was calculated from 350 normal subjects (212 children and 138 adults). The ROC plot showed that the best IGF-I SDS cut-off line was -1.65 for children [sensitivity (S), 68%; specificity (Sp), 97%, diagnostic efficiency (DEf), 81%], the cut-off line for AGHD was -1.65 for AGHD-CO (S, 91%; Sp, 100%; DEf, 94%), and the cut-off line for AGHD-AO was -1.80 (S, 81%; Sp, 100%; DEf, 88%). For IGFBP-3 SDS, the best cut-off line was -1.80 for children (S, 90%; Sp, 60%; DEf, 78%); it was -1.45 for AGHD-CO (S, 90%; Sp, 75%; DEf, 82%) and -0.90 for AGHD-AO (S, 90%; Sp, 68%; DEf, 77%). An accurate diagnosis was obtained using IGF-I SDS alone in GHD children 65%; ISS, 97%; AGHD-CO, 92%; AGHD-AO, 86%, with IGFBP-3 SDS alone in GHD children 60%; ISS, 90%; AGHD-CO, 75%; AGHD-AO, 68%. Considering both, an accurate diagnosis was obtained in GHD children 60%; ISS, 87%; AGHD-CO, 71%; AGHD-AO, 64%. In conclusion, our findings support the need to use cut-off lines expressed in SDS obtained using an appropriate statistical methodology for better characterization of the various clinical presentations. IGF-I proved to be more useful because of its good diagnostic efficiency and accuracy in both children and adults, whereas IGFBP-3 did not significantly contribute to the diagnosis of GHD.


Inhibition of Osteocyte Membrane Repair Activity via Dietary Vitamin E Deprivation Impairs Osteocyte Survival.
Osteocytes experience plasma membrane disruptions (PMD) that initiate mechanotransduction both in vitro and in vivo in response to mechanical loading, suggesting that osteocytes use PMD to sense and adapt to mechanical stimuli. PMD repair is crucial for cell survival; antioxidants (e.g., alpha-tocopherol, also known as Vitamin E) promote repair while reactive oxygen species (ROS), which can accumulate during exercise, inhibit repair. The goal of this study was to determine whether depleting Vitamin E in the diet would impact osteocyte survival and bone adaptation with loading. Male CD-1 mice (3 weeks old) were fed either a regular diet (RD) or Vitamin E-deficient diet (VEDD) for up to 11 weeks. Mice from each dietary group either served as sedentary controls with normal cage activity, or were subjected to treadmill exercise (one bout of exercise or daily exercise for 5 weeks). VEDD-fed mice showed more PMD-affected osteocytes (+ 50%) after a single exercise bout suggesting impaired PMD repair following Vitamin E deprivation. After 5 weeks of daily exercise, VEDD mice failed to show an exercise-induced increase in osteocyte PMD formation, and showed signs of increased osteocytic oxidative stress and impaired osteocyte survival. Surprisingly, exercise-induced increases in cortical bone formation rate were only significant for VEDD-fed mice. This result may be consistent with previous studies in skeletal muscle, where myocyte PMD repair failure (e.g., with muscular dystrophy) initially triggers hypertrophy but later leads to widespread degeneration. In vitro, mechanically wounded MLO-Y4 cells displayed increased post-wounding necrosis (+ 40-fold) in the presence of H2O2, which could be prevented by Vitamin E pre-treatment. Taken together, our data support the idea that antioxidant-influenced osteocyte membrane repair is a vital aspect of bone mechanosensation in the osteocytic control of PMD-driven bone adaptation.


Use of the cauliflower Or gene for improving crop nutritional quality.
Carotenoids are a group of pigments that are essential to human diets. An increasing interest in carotenoids as a nutritional source of vitamin A and health-promoting compounds has prompted the recent progress in metabolic engineering of carotenogenesis in food crops. Current strategies have been mainly focused on manipulating genes encoding carotenogeic enzymes. In many cases, it is difficult to reach the desired levels of carotenoid enhancement. In this chapter, we briefly summarize the recent progress on our understanding of carotenoid biosynthesis. We describe the isolation of a novel gene, the Or gene, from a high-beta-carotene orange cauliflower mutant. The Or gene encodes a plastid-targeted protein containing a cysteine-rich zinc finger domain and appears to be plant-specific. The insertion of a copia-like LTR retrotransponson in the Or gene confers high levels of carotenoid accumulation in the normally low-pigmented tissues. Rather than directly regulating carotenoid biosynthesis, the Or gene controls carotenoid accumulation by inducing the formation of chromoplasts, which provide a metabolic sink to sequester and deposit carotenoids. Examination of the Or transgenic potato tubers confirms that the Or-induced carotenoid accumulation is associated with the formation of a metabolic sink. Thus, the Or gene offers a new molecular tool to complement current approaches for nutritional enhancement in agriculturally important crops.


Possible involvement of pregnane X receptor-enhanced CYP24 expression in drug-induced osteomalacia.
Vitamin D controls calcium homeostasis and the development and maintenance of bones through vitamin D receptor activation. Prolonged therapy with rifampicin or phenobarbital has been shown to cause vitamin D deficiency or osteomalacia, particularly in patients with marginal vitamin D stores. However, the molecular mechanism of this process is unknown. Here we show that these drugs lead to the upregulation of 25-hydroxyvitamin D(3)-24-hydroxylase (CYP24) gene expression through the activation of the nuclear receptor pregnane X receptor (PXR; NR1I2). CYP24 is a mitochondrial enzyme responsible for inactivating vitamin D metabolites. CYP24 mRNA is upregulated in vivo in mice by pregnenolone 16alpha-carbonitrile and dexamethasone, 2 murine PXR agonists, and in vitro in human hepatocytes by rifampicin and hyperforin, 2 human PXR agonists. Moreover, rifampicin increased 24-hydroxylase activity in these cells, while, in vivo in mice, pregnenolone 16alpha-carbonitrile increased the plasma concentration of 24,25-dihydroxyvitamin D(3). Transfection of PXR in human embryonic kidney cells resulted in rifampicin-mediated induction of CYP24 mRNA. Analysis of the human CYP24 promoter showed that PXR transactivates the sequence between -326 and -142. We demonstrated that PXR binds to and transactivates the 2 proximal vitamin D-responsive elements of the human CYP24 promoter. These data suggest that xenobiotics and drugs can modulate CYP24 gene expression and alter vitamin D(3) hormonal activity and calcium homeostasis through the activation of PXR.


Smartphone Applications to Support Tuberculosis Prevention and Treatment: Review and Evaluation.
Tuberculosis (TB) remains a major global health problem and is the leading killer due to a single infectious disease. Mobile health (mHealth)-based tools such as smartphone apps have been suggested as tools to support TB control efforts (eg, identification, contact tracing, case management including patient support). The purpose of this review was to identify and assess the functionalities of mobile apps focused on prevention and treatment of TB. We searched 3 online mobile app stores. Apps were included if they were focused on TB and were in English, Spanish, or Portuguese. For each included app, 11 functionalities were assessed (eg, inform, instruct, record), and searches were conducted to identify peer-review publications of rigorous testing of the available apps. A total of 1332 potentially relevant apps were identified, with 24 meeting our inclusion criteria. All of the apps were free to download, but 7 required login and password and were developed for specific clinics, regional sites, or research studies. Targeted users were mainly clinicians (n=17); few (n=4) apps were patient focused. Most apps (n=17) had 4 or fewer functions out of 11 (range 1-6). The most common functionalities were inform and record (n=15). Although a number of apps were identified with various functionalities to support TB efforts, some had issues such as incorrect spelling and grammar, inconsistent responses to data entry, problems with crashing, or links to features that had no data. Of more concern, some apps provided potentially harmful information to patients, such as links to natural remedies for TB and natural healers. One-third of the apps (8/24) had not been updated for more than a year and may no longer be supported. Peer-reviewed publications were identified for only two of the included apps. In the gray literature (not found in the app stores), three TB-related apps were identified as in progress, being launched, or tested. Apps identified for TB prevention and treatment had minimal functionality, primarily targeted frontline health care workers, and focused on TB information (eg, general information, guidelines, and news) or data collection (eg, replace paper-based notification or tracking). Few apps were developed for use by patients and none were developed to support TB patient involvement and management in their care (eg, follow-up alerts/reminders, side effects monitoring) or improve interaction with their health care providers, limiting the potential of these apps to facilitate patient-centered care. Our evaluation shows that more refined work is needed to be done in the area of apps to support patients with active TB. Involving TB patients in treatment in the design of these apps is recommended.


De novo sequence assembly and characterization of the floral transcriptome in cross- and self-fertilizing plants.
The shift from cross-fertilization to predominant self-fertilization is among the most common evolutionary transitions in the reproductive biology of flowering plants. Increased inbreeding has important consequences for floral morphology, population genetic structure and genome evolution. The transition to selfing is usually characterized by a marked reduction in flower size and the loss of traits involved in pollinator attraction and the avoidance of self-fertilization. Here, we use short-read sequencing to assemble, de novo, the floral transcriptomes of three genotypes of Eichhornia paniculata, including an outcrosser and two genotypes from independently derived selfers, and a single genotype of the sister species E. paradoxa. By sequencing mRNA from tissues sampled at various stages of flower development, our goal was to sequence and assemble the floral transcriptome and identify differential patterns of gene expression. Our 24 Mbp assembly resulted in ~27,000 contigs that averaged ~900 bp in length. All four genotypes had highly correlated gene expression, but the three E. paniculata genotypes were more correlated with one another than each was to E. paradoxa. Our analysis identified 269 genes associated with floral development, 22 of which were differentially expressed in selfing lineages relative to the outcrosser. Many of the differentially expressed genes affect floral traits commonly altered in selfing plants and these represent a set of potential candidate genes for investigating the evolution of the selfing syndrome. Our study is among the first to demonstrate the use of Illumina short read sequencing for de novo transcriptome assembly in non-model species, and the first to implement this technology for comparing floral transcriptomes in outcrossing and selfing plants.


The dual role of paramagnetic particles for integrated lysis and measurement in a rapid immunoassay for intracellular proteins.
A novel, integrated lysis and immunoassay methodology and system for intracellular protein measurement are described. The method uses paramagnetic particles both as a lysis agent and assay label resulting in a rapid test requiring minimal operator intervention, the test being homogeneous and completed in less than 10 min. A design study highlights the critical features of the magnetic detection system used to quantify the paramagnetic particles and a novel frequency-locked loop-based magnetometer is presented. A study of paramagnetic particle enhanced lysis demonstrates that the technique is more than twice as efficient at releasing intracellular protein as ultrasonic lysis alone. Results are presented for measurements of intracellular prostate specific antigen in an LNCAP cell line. This model was selected to demonstrate the rapidity and efficiency of intracellular protein quantification. It was shown that, on average, LNCAP cells contained 0.43 fg of prostate specific antigen. This system promises an attractive solution for applications that require a rapid determination of intracellular proteins.


Subunit-specific analysis of the human immunodeficiency virus type 1 reverse transcriptase in vivo.
The human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) is a heterodimer comprised of two structurally distinct subunits (p51 and p66). Since p51 and p66 are derived from the same coding region, subunit-specific structure-function studies of RT have been conducted exclusively by in vitro biochemical approaches. To study RT subunit function in the context of infectious virus, we constructed an LTR-vpr-p51-IRES-p66 expression cassette in which the HIV-1 vpr gene was fused in frame with p51, followed by an internal ribosome entry site (IRES) sequence and the p66 coding region. By coexpression with RT-deficient proviral DNA, we demonstrated that the p66 subunit is specifically and selectively packaged into virions as a Vpr-p51/p66 complex. Our analysis showed that cleavage by the viral protease liberates Vpr and generates functional heterodimeric RT (p51/p66) that supports HIV-1 reverse transcription and virus infection. By exploiting this novel trans-complementation approach, we demonstrated, for the first time with infectious virions, that the YMDD aspartates of p66 are both required and sufficient for RT polymerase function. Mutational analyses of the p51 YMDD aspartates indicated that they play an important structural role in p51 folding and subunit interactions that are required for the formation of an active RT heterodimer within infected cells. Understanding the role of the individual RT subunits in RNA- and DNA-dependent DNA synthesis is integral to our understanding of RT function. Our findings will lead to important new insights into the role of the p51 and p66 subunits in HIV-1 reverse transcription.


Safety studies of post-surgical buprenorphine therapy for mice.
The use of appropriate analgesia in laboratory mice may be suboptimal because of concerns about adverse events (AE). Target Animal Safety trials were conducted to determine the safety of an extended-release suspension of buprenorphine. Drug or control suspensions were injected subcutaneously in surgically-treated BALB/c mice anesthetized with ketamine-xylazine to mimic post-operative conditions in which the compound might commonly be administered. Single and repeat five-fold (5×) excesses of the 3.25 mg/kg intended dose were used to provoke potential AE. Trials included prospective measurements of weight changes, blood chemistry, hematology, and histopathology. Clinical and histopathology findings were similar in drug-treated and control mice in a four-day trial using a single 16.25 mg/kg, 5× overdose of the drug. In a 12-day trial, which used a total buprenorphine dose of 48.75 mg/kg, clinical and histopathology values were also similar in control and drug-treated female mice. In the male arm of the repeat-overdose trial, two of eight mice died on the morning of day 12, three days following the third 16.25 mg/kg overdose administration. Histopathology did not reveal a cause of death. In a 14-month trial using a single 3.25 mg/kg dose of the drug, no significant findings identified potential AE. These findings indicate a high tolerance to an extended-release buprenorphine suspension administered post-operatively in mice with appropriate husbandry.


The effect of citizenship on the long-term earnings of marginalized immigrants: Quasi-experimental evidence from Switzerland.
We provide evidence that citizenship catalyzes the long-term economic integration of immigrants. Despite the relevance of citizenship policy to immigrant integration, we lack a reliable understanding of the economic consequences of acquiring citizenship. To overcome nonrandom selection into naturalization, we exploit the quasi-random assignment of citizenship in Swiss municipalities that held referendums to decide the outcome of individual naturalization applications. Our data combine individual-level referendum results with detailed social security records from the Swiss authorities. This approach allows us to compare the long-term earnings of otherwise similar immigrants who barely won or lost their referendum. We find that winning Swiss citizenship in the referendum increased annual earnings by an average of approximately 5000 U.S. dollars over the subsequent 15 years. This effect is concentrated among more marginalized immigrants.


Z-scan technique for investigation of the noninstantaneous optical Kerr nonlinearity.
By treating laser-induced optical Kerr nonlinearity as a noninstantaneous decaying process, we present the pulse-duration-dependent Z-scan analytical expressions for an arbitrary aperture and an arbitrary nonlinear magnitude. This theory has the capacity to characterize the third-order nonlinear refraction induced by a laser pulse with its temporal duration being much longer than or comparable to the recovery time of the nonlinear effect. Through Z-scan measurements at different pulse durations, the nonlinear refractive coefficient and the recovery time could be determined unambiguously and simultaneously. Furthermore, the theory can be utilized to confirm whether the measured optical Kerr nonlinearity is instantaneous or noninstantaneous with respect to the given pulse duration.


Self-reported waist circumference: a screening tool for classifying children with overweight/obesity and cardiometabolic risk factor clustering.
The objectives of this study were twofold - (i) to assess the agreement between self-reported waist circumference (SRWC) and assessor measured waist circumference (MWC) and (ii) to evaluate the diagnostic ability of SRWC for classifying (i) a clustering of cardiometabolic risk factors (CMRFs) and (ii) overweight/obese status in Hong Kong Chinese children aged 6-18 years. A cross-sectional study with cluster random sampling was conducted. A self-administrated questionnaire, which included demographic data, body weight, body height and waist circumference, was given to children to bring home for completion. Children were asked to return the questionnaire and fast themselves for at least 8 h on the day of the survey. Anthropometric measurements and blood pressure were taken by trained research staff and fasting blood samples were collected for measurements of fasting plasma glucose, total cholesterol, triglyceride, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol. A total of 515 boys and 711 girls were included in the data analysis. Agreement between SRWC and MWC was assessed by intra-class correlation coefficient and it ranged from 0.77 to 0.87. The ability of sex-specific SRWC values to classify children with a clustering of CMRFs and overweight/obesity exhibited moderately high to high sensitivity and specificity, and the area under the receiver operating characteristics ranged from acceptable to excellent (from 0.76 to 0.84). SRWC has good agreement with MWC and could be used as a screening tool to classify children with a clustering of CMRFs and overweight/obesity status in Hong Kong Chinese children.


Nutrient-based dietary patterns and laryngeal cancer: evidence from an exploratory factor analysis.
The issue of diet and laryngeal cancer has been rarely addressed considering the potential role of dietary patterns. We examined this association using data from a case-control study carried out between 1992 and 2000. Cases were 460 histologically confirmed incident laryngeal cancers hospitalized in two Italian areas. Controls were 1,088 subjects hospitalized for acute nonneoplastic diseases unrelated to tobacco or alcohol consumption. Dietary habits were investigated through a 78-item food frequency questionnaire. A posteriori dietary patterns were identified through principal component factor analysis carried out on a selected set of 28 major nutrients. The internal reproducibility, robustness, and reliability of the identified patterns were evaluated. Odds ratios (OR) of laryngeal cancer and 95% confidence intervals (95% CI) were estimated using unconditional multiple logistic regression models on quartiles of factor scores. We identified five major dietary patterns named "animal products," "starch-rich," "vitamins and fiber," "vegetable unsaturated fatty acids," and "animal unsaturated fatty acids." The vitamins and fiber dietary pattern was inversely associated with laryngeal cancer (OR, 0.35; 95% CI, 0.24-0.52 for the highest versus the lowest score quartile), whereas the animal products (OR, 2.34; 95% CI, 1.59-3.45) and the animal unsaturated fatty acids (OR, 2.07; 95% CI, 1.42-3.01) patterns were directly associated with it. There was no significant association between the vegetable unsaturated fatty acids and the starch-rich patterns and laryngeal cancer risk. These findings suggest that diets rich in animal products and animal fats are directly related, and those rich in fruit and vegetables inversely related, to laryngeal cancer risk.


[First Experience of Clinical Application of LCZ696--an AT1-angiotensin Receptors and Neprilysin Inhibitor--in Patients With Chronic Heart Failure and Reduced Ejection Fraction].
Simultaneous inhibition of the renin-angiotensin-aldosterone system and the system of degradation of natriuretic peptides can potentially provide unique therapeutic effects in patients with chronic heart failure (CHF) with reduced ejection fraction (EF). Aim of this study was to assess tolerability of therapy with LCZ696--first representative of a class of inhibitors of angiotensin receptor and neutral endopeptidase neprilysin--and to study its pharmacodynamic effects. We included into open uncontrolled study 30 patients with stable functional class II-III CHF and EF ≤ 40%. After 24-hour run-in period during which angiotensin converting enzyme inhibitors (ACEI) were withdrawn the patients were given LCZ696 (100 mg/day for 7 days followed by 200 mg/day for 14 days). Other CHF therapy remained unchanged. Transition from therapy with ACEI to LCZ696 was well tolerated. Three patients were excluded because of hyperkalemia ≥ 5mmol/l. After 21 days of treatment elevation of plasma biomarkers of inhibition of neprilysin and angiotensin receptors occurred: cyclic guanosine monophosphate, renin concentration and activity rose 1.38, 3.50, and 2.27 times from baseline level (p < 0.05 for all). After 7 and 21 days of LCZ696 administration we noted significant lowering of NT-proBNP; significant lowering of aldosterone and endothelin-1 in blood plasma, was observed on day 21. Administration of LCZ696 to patients with CHF with reduced ejection fraction (EF) was well tolerated and associated with potentially favorable for this category of patients dynamics of biomarkers.


Common variants near MC4R: exploring gender effects in overweight and obese children and adolescents participating in a lifestyle intervention.
Association with obesity and increased insulin levels have been reported for two variants (rs17782313 and rs12970134) located downstream of the melanocortin-4 receptor gene (MC4R). We investigated whether these variants have sex-specific effects on overweight, obesity and 14 related phenotypes in 889 overweight and obese children and adolescents. We also explored the impact of the variants on weight change in 367 of the 889 cases who participated in an intervention program. Prior to these analyses we showed that both variants were associated with overweight/obesity in the analyzed 889 cases versus 442 normal-weight and lean controls (case-control study). In explorative analyses we observed higher diastolic blood pressure levels in males (rs17782313: β = 2.52 mm Hg per risk allele; p = 0.003) but reduced blood pressure level in females for the same risk allele (β = -1.72 mm Hg; p = 0.039). We also detected a greater BMI standard deviation score (BMI-SDS) reduction in females with the risk allele at rs17782313 (β = 0.086 per risk allele; p = 0.021). Additionally, we observed evidence for an association of the same risk allele with insulin levels (β = 0.029 log (μU/ml); p = 0.044) with no sex-specific effect. For the remaining 11 phenotypes, we observed no evidence for a (sex-specific) association. In sum, our data support the associations of variants rs17782313 and rs12970134 near MC4R with early onset obesity and increased insulin levels. Exploratory evidence for sex-specific effects of the risk alleles were observed for diastolic blood pressure and BMI-SDS reduction.


T cell development and T cell responses in mice with mutations affecting tyrosines 292 or 315 of the ZAP-70 protein tyrosine kinase.
After stimulation of the T cell receptor (TCR), the tyrosine residues 292 and 315 in interdomain B of the protein tyrosine kinase ZAP-70 become phosphorylated and plausibly function as docking sites for Cbl and Vav1, respectively. The two latter proteins have been suggested to serve as substrates for ZAP-70 and to fine-tune its function. To address the role of these residues in T cell development and in the function of primary T cells, we have generated mice that express ZAP-70 molecules with Tyr to Phe substitution at position 292 (Y292F) or 315 (Y315F). When analyzed in a sensitized TCR transgenic background, the ZAP-70 Y315F mutation reduced the rate of positive selection and delayed the occurrence of negative selection. Furthermore, this mutation unexpectedly affected the constitutive levels of the CD3-zeta p21 phosphoisoform. Conversely, the ZAP-70 Y292F mutation upregulated proximal events in TCR signaling and allowed more T cells to produce interleukin 2 and interferon gamma in response to a given dose of antigen. The observation that ZAP-70 Y292F T cells have a slower rate of ligand-induced TCR downmodulation suggests that Y292 is likely involved in regulating the duration activated TCR reside at the cell surface. Furthermore, we showed that Y292 and Y315 are dispensable for the TCR-induced tyrosine phosphorylation of Cbl and Vav1, respectively. Therefore, other molecules present in the TCR signaling cassette act as additional adaptors for Cbl and Vav1. The present in vivo analyses extend previous data based on transformed T cell lines and suggest that residue Y292 plays a role in attenuation of TCR signaling, whereas residue Y315 enhances ZAP-70 function.


The diagnosis of diverticulitis in outpatients: on what evidence?
Diverticular disease is common in the outpatient setting; yet, rigorous study of diagnosis and management strategies is currently limited to hospitalized patients. Here, we characterize the clinical assessment generating the diagnostic label of diverticulitis in outpatients. Encounters for diverticulitis were identified using ICD-9 diagnosis codes (562.11/562.13) from the electronic medical record system of a tertiary referral hospital and its regional clinics. The frequencies of various demographic and clinical variables were compared between patients presenting in the emergency room (ER) or outpatient Clinic. Between 2003 and 2008, 820 inpatients and 2,576 outpatients met inclusion criteria (328 [13%] ER, 2,248 [87%] Clinic). Compared to ER patients, Clinic patients were less likely to undergo urgent abdominal/pelvic computed tomography (CT) scan (14% vs. 85%, p<.0001) or have an abnormal WBC count (35% vs. 69%, p<.0001). Twenty-four hour , including inpatient admission (30% ER vs. 3.5% Clinic, p<.0001) and colectomy (1.2% ER vs. 0.4% Clinic, p=0.08) were rare in both groups. Diverticulitis in the outpatient setting is often characterized by infrequent use of CT scans, lack of leukocytosis, and rare need for urgent surgery or early admission. As this diagnostic label appears to be commonly applied without objective evidence, further study is needed to evaluate its validity.


A data-driven computational scheme for the nonlinear mechanical properties of cellular mechanical metamaterials under large deformation.
Cellular mechanical metamaterials are a special class of materials whose mechanical properties are primarily determined by their geometry. However, capturing the nonlinear mechanical behavior of these materials, especially those with complex geometries and under large deformation, can be challenging due to inherent computational complexity. In this work, we propose a data-driven multiscale computational scheme as a possible route to resolve this challenge. We use a neural network to approximate the effective strain energy density as a function of cellular geometry and overall deformation. The network is constructed by "learning" from the data generated by finite element calculation of a set of representative volume elements at cellular scales. This effective strain energy density is then used to predict the mechanical responses of cellular materials at larger scales. Compared with direct finite element simulation, the proposed scheme can reduce the computational time up to two orders of magnitude. Potentially, this scheme can facilitate new optimization algorithms for designing cellular materials of highly specific mechanical properties.


Platelet Lipidomic Profiling: Novel Insight into Cytosolic Phospholipase A2α Activity and Its Role in Human Platelet Activation.
With a newer, more selective and efficacious cytosolic phospholipase A2α (cPLA2α) inhibitor available, we revisited the role of cPLA2α activity in platelet activation and discovered that a component of platelet signaling, even larger than previously appreciated, relies on this enzyme. In a whole blood shear-based flow chamber assay, giripladib, a cPLA2α inhibitor, reduced platelet adhesion and accumulation on collagen. Moreover, giripladib differentially affected P-selectin expression and GPIIbIIIa activation depending on the agonist employed. While protease-activated receptor 1 (PAR1)-mediated platelet activation was unaffected by giripladib, the levels of PAR4- and GPVI-mediated platelet activation were significantly reduced. Meanwhile, the thromboxane A2 receptor antagonist SQ29548 had no effect on PAR-, GPVI-, or puriniergic receptor-mediated platelet activation, suggesting that another eicosanoid produced downstream of arachidonic acid liberation by cPLA2α was responsible for this large component of PAR4- and GPVI-mediated platelet activation. In parallel, we profiled PAR-mediated changes in glycerophospholipid (GPL) mass with and without giripladib to better understand cPLA2α-mediated lipid metabolism. Phosphatidylcholine and phosphatidylethanolamine (PE) demonstrated the largest consumption of mass during thrombin stimulation. Additionally, we confirm phosphatidylinositol as a major substrate of cPLA2α. A comparison of PAR1- and PAR4-induced metabolism revealed the consumption of more putative arachidonyl-PE species downstream of PAR1 activation. Instead of enhanced cPLA2α activity and therefore more arachidonic acid liberation downstream of PAR4, these results indicate the major role that cPLA2α activity plays in platelet function and suggest that a novel eicosanoid is produced in response to platelet activation that represents a large component of PAR4- and GPVI-mediated responses.


Identification of a candidate CD5 homologue in the amphibian Xenopus laevis.
We identified a novel T cell Ag in the South African clawed toad (Xenopus laevis) by a mAb designated 2B1. This Ag is present in relatively high levels on most thymocytes, approximately 65% of splenocytes, 55% of PBL, and 65% of intestinal lymphocytes, but is rarely seen on IgM+ B cells in any of these tissues. Lymphocytes bearing the 2B1 Ag proliferate in response to stimulation with Con A or PHA, whereas the 2B1- lymphocytes are reactive to LPS. Biochemical analysis indicates that this Ag is a differentially phosphorylated glycoprotein of 71 to 82 kDa. The protein core of 64 kDa bears both N- and O-linked carbohydrate side chains. The amino-terminal protein sequence of the 2B1 Ag shares significant homology with both the macrophage scavenger receptor type 1 motif and the mammalian CD5/CD6 family. The biochemical characteristics and cellular distribution of the 2B1 Ag suggest that it represents the CD5 homologue in X. laevis. While T cells constitutively express this highly conserved molecule, Xenopus B cells acquire the CD5 homologue only when they are stimulated in the presence of T cells.


Lasting effects of climate disturbance on perennial grassland above-ground biomass production under two cutting frequencies.
Climate extremes can ultimately reshape grassland services such as forage production and change plant functional type composition. This 3-year field research studied resistance to dehydration and recovery after rehydration of plant community and plant functional types in an upland perennial grassland subjected to climate and cutting frequency (Cut+, Cut-) disturbances by measuring green tissue percentage and above-ground biomass production (ANPP). In year 1, a climate disturbance gradient was applied by co-manipulating temperature and precipitation. Four treatments were considered: control and warming-drought climatic treatment, with or without extreme summer event. In year 2, control and warming-drought treatments were maintained without extreme. In year 3, all treatments received ambient climatic conditions. We found that the grassland community was very sensitive to dehydration during the summer extreme: aerial senescence reached 80% when cumulated climatic water balance fell to -156 mm and biomass declined by 78% at the end of summer. In autumn, canopy greenness and biomass totally recovered in control but not in the warming-drought treatment. However ANPP decreased under both climatic treatments, but the effect was stronger on Cut+ (-24%) than Cut- (-15%). This decline was not compensated by the presence of three functional types because they were negatively affected by the climatic treatments, suggesting an absence of buffering effect on grassland production. In the following 2 years, lasting effects of climate disturbance on ANPP were observable. The unexpected stressful conditions of year 3 induced a decline in grassland production in the Cut+ control treatment. The fact that this treatment cumulated higher (45%) N export over the 3 years suggests that N plays a key role in ANPP stability. As ANPP in this mesic perennial grassland did not show engineering resilience, long-term experimental manipulation is needed. Infrequent mowing appears more appropriate for sustaining grassland ANPP under future climate extremes.