Improving human hemato-lymphoid-system mice by cytokine knock-in gene replacement.
Human hemato-lymphoid-system mice hold great promise for modeling and studying important human diseases in vivo, and to enable vaccine testing. Until now, several major limitations have restricted the utility of human hemato-lymphoid-system mice in translational research. Recently, however, significant advances have been made in improving these mice, based on the delivery of human cytokines to create a better environment for human cells in the mouse host. In this review, we discuss the various approaches with a particular focus on improving human hemato-lymphoid-system mice by human cytokine knock-in gene replacement.


Risk of second primary cancer following myeloid neoplasia and risk of myeloid neoplasia as second primary cancer: a nationwide, observational follow up study in Sweden.
Although advances in the treatment of myeloid neoplasms have led to improved patient survival, this improvement has been accompanied by an increased risk of second primary cancer (ie, the risk of another cancer after myeloid neoplasia). We aimed to assess bi-directional associations between myeloid cancers and other cancers-ie, development of second primary cancer in patients who have previously had myeloid cancer, and risks of myeloid neoplasia in patients who have previously had another cancer-to provide insight into possible mechanisms beyond side-effects of treatment and shared risk factors. Using the Swedish Family-Cancer Database, we identified 35 928 individuals with primary myeloid cancer, including myeloproliferative neoplasms, acute myeloid leukaemia, chronic myeloid leukaemia, and myelodysplastic syndrome diagnosed between 1958 and 2015. The Swedish Family-Cancer Database includes every individual registered as a resident in Sweden starting in 1932, with full parental history. The primary endpoint was the assessment of relative risks (RRs) for second primary cancer, which we performed using means of incidence rate ratios, regressed over a generalised Poisson model. Between 1958 and 2015, overall relative risk of second primary cancers was significantly increased after acute myeloid leukaemia (RR 1·29, 95% CI 1·17-1·41), chronic myeloid leukaemia (1·52, 1·35-1·69), myelodysplastic syndrome (1·42, 1·26-1·59), and all myeloproliferative neoplasms (1·37, 1·30-1·43) relative to the incidence of these cancers as first primary cancer. With myeloid neoplasia as a second primary cancer, risks were significantly increased for acute myeloid leukaemia (1·57, 1·48-1·65), chronic myeloid leukaemia (1·26, 1·13-1·40), and myelodysplastic syndrome (1·54, 1·42-1·67) relative to the incidence of these myeloid neoplasms as first primary cancers. Relative risk of upper aerodigestive tract cancer, squamous cell skin cancer, and non-Hodgkin lymphoma as second primary cancers were increased after all four types of myeloid neoplasia relative to their incidence as first primary cancers. High risks of myelodysplastic syndrome and acute myeloid leukaemia as second primary cancers were found after haematological cancers (RRs between 5·08 and 10·04). The relative risks of second primary cancer are important for the long-term management of patients with myeloid cancers. The bi-directional associations of myeloid cancers with many other cancers suggest a number of candidate mechanisms that might contribute to the development and aetiology of a second primary cancer. These mechanisms might include immune dysfunction or the effects of treatment, and these should be assessed in future investigations. Deutsche Krebshilfe, Jane and Aatos Erkko Foundation, Sigrid Juselius Foundation, Finnish Cancer Organizations, Swedish Research Council, ALF from Region Skåne, and Bloodwise.


Rapid N-acetyltransferase 2 imputed phenotype and smoking may increase risk of colorectal cancer in women (Netherlands).
The relationship between smoking and colorectal cancer risk and whether such effect is modified by variations in the NAT2 genotype is investigated. In the prospective DOM (Diagnostisch Onderzoek Mammacarcinoom; 27,722 women) cohort follow-up from 1976 until 1987 revealed 54 deaths due to colon or rectal cancer, and follow-up from 1987 to 01-01-1996 revealed 204 incident colorectal cancer cases. A random sample (n = 857) from the baseline cohort was used as controls. Four NAT2 restriction fragment length polymorphisms (RFLPs) were analysed using DNA extracted from urine samples. Rapid or slow acetylator phenotype status was attributed to individuals. Smoking may increase the risk for colon cancer (RR = 1.36, 95% CI 0.97-1.92) as well as for rectal cancer (RR = 1.31, 95% CI 0.76-2.25), although not statistically significant. Rapid NAT2 acetylation did not increase colorectal cancer risk, but in combination with smoking the risk was statistically significant increased, compared to women who had a slow NAT2 imputed phenotype and never smoked (RR = 1.56, 95% CI 1.03-2.37). For colon cancer, but not for rectal cancer the increased risk was statistically significant (RR = 1.67, 95% CI, 1.05-2.67 versus RR = 1.30 95% CI 0.63-2.68). Our study points to smoking as a risk factor for colon and rectal cancer and, in addition, especially in women with rapid NAT2 imputed phenotype.


Cannabinoid hyperemesis.
A 21-year-old woman presented with a 4-week history of sudden onset vomiting, nausea and anorexia. Questioning revealed that she had a 7-year history of heavy cannabis use (smoking). She did not describe abdominal pain, change in bowel habit, antibiotic use, foreign travel or contact with gastroenteritis. Biochemistry results demonstrated mild metabolic derangement with a low potassium and a low bicarbonate, and urine toxicology was positive for cannabinoids. Other investigations, including a full blood count, renal function tests, liver function tests, a coagulation sample, an ECG, urinary β-hCG and a CT head scan, were all normal. A diagnosis of cannabinoid hyperemesis was made and her symptoms resolved after treatment with intravenous fluids, antiemetics and abstinence from cannabis. Since her discharge and abstinence she has had several relapses, each related to cannabis use and each resolving with abstinence. The patient is now seeking cognitive behavioural therapy to achieve permanent abstinence.


Special educators' knowledge and training on autism in Karnataka: A cross-sectional study.
In the wake of increasing prevalence of autism globally, with a substantial change in understanding of the disorder, it is essential to update knowledge of the healthcare professionals involved in the intervention of children with autism. Special educators are important team members in the rehabilitation of children with autism. There are only a few studies addressing knowledge and training of special educators in the rehabilitation of children with autism, particularly in the Indian context. The present study investigated knowledge and training on autism among special educators in a southern state of India, Karnataka. A descriptive cross-sectional study design was adopted for this study. Data were collected from 47 special educators, who answered questions related to general knowledge, knowledge of educational programming, knowledge about classroom behaviors, and professional development needs regarding autism. Statistical analysis of the data was performed using Statistical Package for Social Sciences (version 16). Results of the study indicated that special educators had an overall reduced level of understanding about autism and poor knowledge on educational programming and classroom behaviors in children with autism. Further, their knowledge regarding autism varied with educational qualification and years of work experience. Findings of the study, thus, emphasize the need for increasing awareness by providing knowledge and training to special educators working with children with autism in India.


Endothelial-specific intron-derived miR-126 is down-regulated in human breast cancer and targets both VEGFA and PIK3R2.
Endothelial cells are the key components of vascular intima and play pivotal roles in vasculogenesis, angiogenesis, and tumor growth. Using Northern blot and real-time PCR, we confirmed that miR-126 and its host gene EGF-like domain 7 (EGFL7) were widely expressed in rat tissues but strictly expressed in endothelial cells. In mammals, miR-126 gene is embedded in intron7 of EGFL7. To explore the biogenesis of miR-126, plasmid EGFL7(126)-pEGFPc1 containing segment of exon7-intron7-exon8 of EGFL7 was constructed and expressed in 293T. Expression of spliced exon7-8 and excised mature miR-126 was detected by PCR and Northern blot. Knocking-down of endothelial endogenous miR-126 did not affect EGFL7 expression at mRNA or protein level. To investigate the possible roles of miR-126, PicTar, miRBase, miRanda, Bibiserv, and Targetscan were used to screen the targets. VEGFA and PIK3R2 were confirmed as the targets of miR-126 by luciferase reporter assay and Western blot. Interestingly, Northern blot and western blot showed that miR-126 was down-regulated in breast tumors where the VEGF/PI3K/AKT signaling pathway was activated. Introduction of miR-126 mimics into MCF-7 could effectively decrease VEGF/PI3K/AKT signaling activity. In summary, miR-126 was strictly expressed in endothelial cells and excised from EGFL7 pre-mRNA without affecting splicing and expression of its host gene. In addition, miR-126 could target both VEGFA and PIK3R2, and its expression was decreased in human breast cancer, implying that miR-126 may play a role in tumor genesis and growth by regulating the VEGF/PI3K/AKT signaling pathway.


Targeting hepatic glutaminase activity to ameliorate hyperglycemia.
Glucagon levels increase under homeostatic, fasting conditions, promoting the release of glucose from the liver by accelerating the breakdown of glycogen (also known as glycogenolysis). Glucagon also enhances gluconeogenic flux, including from an increase in the hepatic consumption of amino acids. In type 2 diabetes, dysregulated glucagon signaling contributes to the elevated hepatic glucose output and fasting hyperglycemia that occur in this condition. Yet, the mechanism by which glucagon stimulates gluconeogenesis remains incompletely understood. Contrary to the prevailing belief that glucagon acts primarily on cytoplasmic and nuclear targets, we find glucagon-dependent stimulation of mitochondrial anaplerotic flux from glutamine that increases the contribution of this amino acid to the carbons of glucose generated during gluconeogenesis. This enhanced glucose production is dependent on protein kinase A (PKA) and is associated with glucagon-stimulated calcium release from the endoplasmic reticulum, activation of mitochondrial α-ketoglutarate dehydrogenase, and increased glutaminolysis. Mice with reduced levels of hepatic glutaminase 2 (GLS2), the enzyme that catalyzes the first step in glutamine metabolism, show lower glucagon-stimulated glutamine-to-glucose flux in vivo, and GLS2 knockout results in higher fasting plasma glucagon and glutamine levels with lower fasting blood glucose levels in insulin-resistant conditions. As found in genome-wide association studies (GWAS), human genetic variation in the region of GLS2 is associated with higher fasting plasma glucose; here we show in human cryopreserved primary hepatocytes in vitro that these natural gain-of-function missense mutations in GLS2 result in higher glutaminolysis and glucose production. These data emphasize the importance of gluconeogenesis from glutamine, particularly in pathological states of increased glucagon signaling, while suggesting a possible new therapeutic avenue to treat hyperglycemia.


When clinical trials compete: prioritising study recruitment.
It is not uncommon for multiple clinical trials at the same institution to recruit concurrently from the same patient population. When the relevant pool of patients is limited, as it often is, trials essentially compete for participants. There is evidence that such a competition is a predictor of low study accrual, with increased competition tied to increased recruitment shortfalls. But there is no consensus on what steps, if any, institutions should take to approach this issue. In this article, we argue that an institutional policy that prioritises some trials for recruitment ahead of others is ethically permissible and indeed prima facie preferable to alternative means of addressing recruitment competition. We motivate this view by appeal to the ethical importance of minimising the number of studies that begin but do not complete, thereby exposing their participants to unnecessary risks and burdens in the process. We then argue that a policy of prioritisation can be fair to relevant stakeholders, including participants, investigators and funders. Finally, by way of encouraging and helping to frame future debate, we propose some questions that would need to be addressed when identifying substantive ethical criteria for prioritising between studies.


Site-specific activity of the acyltransferases HtrB1 and HtrB2 in Pseudomonas aeruginosa lipid A biosynthesis.
Pseudomonas aeruginosa (PA) is an opportunistic Gram-negative pathogen associated with nosocomial infections, acute infections and chronic lung infections in patients with cystic fibrosis. The ability of PA to cause infection can be attributed to its ability to adapt to a multitude of environments. Modification of the lipid A portion of lipopolysaccharide (LPS) is a vital mechanism Gram-negative pathogens use to remodel the outer membrane in response to environmental stimuli. Lipid A, the endotoxic moiety of LPS, is the major component of the outer leaflet of the outer membrane of Gram-negative bacteria making it a critical factor for bacterial adaptation. One way PA modifies its lipid A is through the addition of laurate and 2-hydroxylaurate. This secondary or late acylation is carried out by the acyltransferase, HtrB (LpxL). Analysis of the PA genome revealed the presence of two htrB homologs, PA0011 (htrB1) and PA3242 (htrB2). In this study, we were able to show that each gene identified is responsible for site-specific modification of lipid A. Additionally, deletions of either gene altered resistance to specific classes of antibiotics, cationic antimicrobial peptides and increased membrane permeability suggesting a role for these enzymes in maintaining optimal membrane organization and integrity.


Characterizing hospital workers' willingness to report to duty in an influenza pandemic through threat- and efficacy-based assessment.
Hospital-based providers' willingness to report to work during an influenza pandemic is a critical yet under-studied phenomenon. Witte's Extended Parallel Process Model (EPPM) has been shown to be useful for understanding adaptive behavior of public health workers to an unknown risk, and thus offers a framework for examining scenario-specific willingness to respond among hospital staff. We administered an anonymous online EPPM-based survey about attitudes/beliefs toward emergency response, to all 18,612 employees of the Johns Hopkins Hospital from January to March 2009. Surveys were completed by 3426 employees (18.4%), approximately one third of whom were health professionals. Demographic and professional distribution of respondents was similar to all hospital staff. Overall, more than one-in-four (28%) hospital workers indicated they were not willing to respond to an influenza pandemic scenario if asked but not required to do so. Only an additional 10% were willing if required. One-third (32%) of participants reported they would be unwilling to respond in the event of a more severe pandemic influenza scenario. These response rates were consistent across different departments, and were one-third lower among nurses as compared with physicians. Respondents who were hesitant to agree to work additional hours when required were 17 times less likely to respond during a pandemic if asked. Sixty percent of the workers perceived their peers as likely to report to work in such an emergency, and were ten times more likely than others to do so themselves. Hospital employees with a perception of high efficacy had 5.8 times higher declared rates of willingness to respond to an influenza pandemic. Significant gaps exist in hospital workers' willingness to respond, and the EPPM is a useful framework to assess these gaps. Several attitudinal indicators can help to identify hospital employees unlikely to respond. The findings point to certain hospital-based communication and training strategies to boost employees' response willingness, including promoting pre-event plans for home-based dependents; ensuring adequate supplies of personal protective equipment, vaccines and antiviral drugs for all hospital employees; and establishing a subjective norm of awareness and preparedness.


Optical fiber probe spectroscopy for laparoscopic monitoring of tissue oxygenation during esophagectomies.
Anastomotic complication is a major morbidity associated with esophagectomy. Gastric ischemia after conduit creation contributes to anastomotic complications, but a reliable method to assess oxygenation in the gastric conduit is lacking. We hypothesize that fiber optic spectroscopy can reliably assess conduit oxygenation, and that intraoperative gastric ischemia will correlate with the development of anastomotic complications. A simple optical fiber probe spectrometer is designed for nondestructive laparoscopic measurement of blood content and hemoglobin oxygen saturation in the stomach tissue microvasculature during human esophagectomies. In 22 patients, the probe measured the light transport in stomach tissue between two fibers spaced 3-mm apart (500- to 650-nm wavelength range). The stomach tissue site of measurement becomes the site of a gastroesophageal anastamosis following excision of the cancerous esophagus and surgical ligation of two of the three gastric arteries that provide blood perfusion to the anastamosis. Measurements are made at each of five steps throughout the surgery. The resting baseline saturation is 0.51±0.15 and decreases to 0.35±0.20 with ligation. Seven patients develop anastomotic complications, and a decreased saturation at either of the last two steps (completion of conduit and completion of anastamosis) is predictive of complication with a sensitivity of 0.71 when the specificity equaled 0.71.


Targeting mitochondrial dysfunction in the treatment of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) - a clinical audit.
We report on an audit of 138 ME/CFS patients who attended a private practice and took the ATP Profile biomedical test. The results revealed that all of these patients had measureable mitochondrial dysfunction. A basic treatment regime, based on 1) eating the evolutionary correct stone-age diet, 2) ensuring optimum hours of good quality sleep, 3) taking a standard package of nutritional supplements, and 4) getting the right balance between work and rest, was recommended for all patients. Additions to the basic regime were tailored for each patient according to the results of the ATP Profile and additional nutritional tests and clues from the clinical history. Mitochondrial function is typically impaired in two ways: substrate or co-factor deficiency, and inhibition by chemicals, exogenous or endogenous. For the former, additional nutrients are recommended where there is a deficiency, and for the latter, improvement of anti-oxidant status and selective chelation therapy or far-infrared saunas are appropriate. We show case histories of nine patients who have taken the ATP Profile on three or four occasions, and a before-and-after treatment summary of the 34 patients who have had at least two ATP Profile tests separated by some months. Finally, we summarize the results for the 30 patients who followed all aspects of the treatment regime and compare them with the 4 patients who were lax on two or more aspects of the treatment regime. All patients who followed the treatment regime improved in mitochondrial function by on average a factor of 4.


Rare copy number variants contribute to congenital left-sided heart disease.
Left-sided congenital heart disease (CHD) encompasses a spectrum of malformations that range from bicuspid aortic valve to hypoplastic left heart syndrome. It contributes significantly to infant mortality and has serious implications in adult cardiology. Although left-sided CHD is known to be highly heritable, the underlying genetic determinants are largely unidentified. In this study, we sought to determine the impact of structural genomic variation on left-sided CHD and compared multiplex families (464 individuals with 174 affecteds (37.5%) in 59 multiplex families and 8 trios) to 1,582 well-phenotyped controls. 73 unique inherited or de novo CNVs in 54 individuals were identified in the left-sided CHD cohort. After stringent filtering, our gene inventory reveals 25 new candidates for LS-CHD pathogenesis, such as SMC1A, MFAP4, and CTHRC1, and overlaps with several known syndromic loci. Conservative estimation examining the overlap of the prioritized gene content with CNVs present only in affected individuals in our cohort implies a strong effect for unique CNVs in at least 10% of left-sided CHD cases. Enrichment testing of gene content in all identified CNVs showed a significant association with angiogenesis. In this first family-based CNV study of left-sided CHD, we found that both co-segregating and de novo events associate with disease in a complex fashion at structural genomic level. Often viewed as an anatomically circumscript disease, a subset of left-sided CHD may in fact reflect more general genetic perturbations of angiogenesis and/or vascular biology.


Minimal memory for details in real life events.
The extent to which the details of past experiences are retained or forgotten remains controversial. Some studies suggest massive storage while others describe memories as fallible summary recreations of original events. The discrepancy can be ascribed to the content of memories and how memories are evaluated. Many studies have focused on recalling lists of words/pictures, which lack the critical ingredients of real world memories. Here we quantified the ability to remember details about one hour of real life. We recorded video and eye movements while subjects walked along specified routes and evaluated whether they could distinguish video clips from their own experience from foils. Subjects were minimally above chance in remembering the minutiae of their experiences. Recognition of specific events could be partly explained by a machine-learning model of video contents. These results quantify recognition memory for events in real life and show that the details of everyday experience are largely not retained in memory.


Visual acuity and egg spatial chromatic contrast predict egg rejection behavior of American robins.
Color and spatial vision is critical for recognition and discrimination tasks affecting fitness, including finding food and mates, and recognizing offspring. For example, as a counter defense to avoid the cost of raising the unrelated offspring of obligate interspecific avian brood parasites, many host species routinely view, recognize and remove the foreign egg(s) from their nests. Recent research has shown that host species visually attend to both chromatic and spatial pattern features of eggs; yet how hosts simultaneously integrate these features together when recognizing eggs remains an open question. Here, we tested egg rejection responses of American robins (Turdus migratorius) using a range of 3D-printed model eggs covered with blue and yellow checkered patterns differing in relative square sizes. We predicted that robins would reject a model egg if they could visually resolve the blue and yellow squares as separate features, or accept it if the squares blended together and appeared similar in color to the natural blue-green color of robin eggs as perceived by the avian visual system. As predicted, the probability of robins rejecting a model egg increased with greater sizes of its blue and yellow squares. Our results suggest that chromatic visual acuity and viewing distance have the potential to limit the ability of a bird to recognize a foreign egg in its nest, thus providing a limitation to host egg recognition that obligate interspecific avian brood parasites may exploit.


Cytotoxic and mutagenic potential of solutions exposed to cold atmospheric plasma.
The exposure of aqueous solutions to atmospheric plasmas results in the generation of relatively long-lived secondary products such as hydrogen peroxide which are biologically active and have demonstrated anti-microbial and cytotoxic activity. The use of plasma-activated solutions in applications such as microbial decontamination or anti-cancer treatments requires not only adequate performance on target cells but also a safe operating window regarding the impact on surrounding tissues. Furthermore the generation of plasma-activated fluids needs to be considered as a by-stander effect of subjecting tissue to plasma discharges. Cytotoxicity and mutagenicity assays using mammalian cell lines were used to elucidate the effects of solutions treated with di-electric barrier discharge atmospheric cold plasma. Plasma-treated PBS inhibited cell growth in a treatment time-dependent manner showing a linear correlation to the solutions' peroxide concentration which remained stable over several weeks. Plasma-treated foetal bovine serum (FBS) acting as a model for complex bio-fluids showed not only cytotoxic effects but also exhibited increased mutagenic potential as determined using the mammalian HPRT assay. Further studies are warranted to determine the nature, causes and effects of the cyto- and genotoxic potential of solutions exposed to plasma discharges to ensure long-term safety of novel plasma applications in medicine and healthcare.


The supercritical CO₂ extract from the skin of Bufo bufo gargarizans Cantor blocks hepatitis B virus antigen secretion in HepG2.2.15 cells.
The skin of Bufo bufo gargarizans Cantor has long been used for the treatment of hepatitis B in China and supercritical carbon dioxide extraction (SC-CO₂) is widely used in extracting active ingredients from natural products. The aim of present study was to assess the anti-hepatitis B virus (HBV) effect of the supercritical CO₂ extract from the skin of Bufo bufo gargarizans Cantor (SCE-BC). Cytotoxicity of SCE-BC was analyzed using an MTT [3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide] assay in HepG2.2.15 cells. The hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg), and hepatitis B core-related antigen (HBcrAg) concentrations in cell culture medium were determined by chemiluminescent enzyme immunoassay. HBV mRNA in cells was determined using real-time polymerase chain reaction. SCE-BC concentrations below 10(-2) μg/mL had no significant toxicity to HepG2.2.15 cells. SCE-BC at 10(-4) μg/mL effectively inhibited the secretion of HBeAg by 23.36% on day 6. It was more potent than the positive control lamivudine (100 μg/mL) in terms of the inhibition of HBeAg and HBcrAg secretion on day 6. Consistent with the HBV antigen reduction, HBV mRNA expression was markedly inhibited in comparison to the control when HepG2.2.15 cells were treated with SCE-BC. Moreover, SCE-BC had greater inhibitory activity with respect to HBeAg than to HBsAg. Since HBeAg promotes immune tolerance and persistent infection during HBV infection, the present results suggest that immune tolerance induced by HBeAg might be overcome by SCE-BC. Therefore, SCE-BC warrants further investigation.


Suitability of the Vero cell method for titration of diphtheria antitoxin in the United States potency test for diphtheria toxoid.
The in vitro Vero cell method for titration of diphtheria antitoxin in immunized guinea pig sera was standardized to obtain comparable results to the in vivo toxin neutralization (TN) test in guinea pigs used to test potency of adsorbed diphtheria toxoid according to the United States Minimum Requirements. In the Vero cell method, the antitoxin titers of the guinea pig sera, obtained 4 weeks after immunization, were markedly dependent on the toxin dose level used in the assay. A toxin dose level termed the Lcd/1 dose (limit of cytopathic dose at 1 IU/ml) for Vero cell method gave comparable estimates of antitoxin activity to the in vivo TN method performed at L+/1 dose of toxin. The Lcd/1 toxin dose level for the Vero cell method is defined as the minimum concentration of diphtheria toxin required to produce a cytopathic effect on Vero cells in 4 days when the solution of diphtheria toxin is mixed with an equal volume of standard diphtheria antitoxin whose concentration is 1 IU/ml. When lower dose levels of toxin were used in the Vero cell assay (Lcd/10 to Lcd/1000), the concentrations of antitoxin in 4 weeks guinea pig sera were 2 to 11.7 times lower than with the Lcd/1 dose level and similarly lower than those measured by the in vivo TN test at L+/1 level. In contrast, the concentration of antitoxin measured in sera from guinea pigs who have been boosted with diphtheria toxoid increased approximately two-fold with the Lcd/1000 dose level.(ABSTRACT TRUNCATED AT 250 WORDS)


Toxic heme in sickle cells: an explanation for death of malaria parasites.
In an effort to elucidate a mechanism of genetic resistance to malaria, we asked whether a toxic form of heme is included in the excess of ferriprotoporphyrin IX (FP) which has been reported to accumulate as hemichromes in sickle cells. When FP is bound to certain erythrocytic elements, such as native hemoglobin, it is inaccessible to bind chloroquine with high affinity and is nontoxic. However, when FP is accessible to bind chloroquine with high affinity, it has been demonstrated to be sufficiently free to have membrane toxicity and, under certain conditions, to lyse Plasmodium falciparum parasites. [14C]-chloroquine was used, therefore, as a reporter molecule to evaluate the quantity, accessibility, and potential toxicity of FP released from hemoglobin. Intact erythrocytes from subjects with sickle cell anemia bound approximately 71 mumoles of chloroquine per kg with an apparent Kd of 10(-6) M. Erythrocytes from normal subjects or subjects with sickle trait bound little or no chloroquine with high affinity. Since the oxidant stress introduced by malaria parasites would increase the tendency for denaturation of hemoglobin S with additional release of FP, we suggest that FP toxicity accounts for the death of malaria parasites in sickle cells.


TACI is mutant in common variable immunodeficiency and IgA deficiency.
The tumor necrosis factor receptor family member TACI (transmembrane activator and calcium-modulator and cyclophilin ligand interactor) mediates isotype switching in B cells. We found that 4 of 19 unrelated individuals with common variable immunodeficiency (CVID) and 1 of 16 individuals with IgA deficiency (IgAD) had a missense mutation in one allele of TNFRSF13B (encoding TACI). One of the four individuals with CVID had a single nucleotide insertion in the other TNFRSF13B allele. None of these mutations were present in 50 healthy subjects. TNFRSF13B mutations cosegregated with the phenotype of CVID or IgAD in family members of four index individuals that we studied. B cells from individuals with TACI mutations expressed TACI but did not produce IgG and IgA in response to the TACI ligand APRIL, probably reflecting impaired isotype switching. These results suggest that TACI mutations can result in CVID and IgAD.


A Transgenic Camelina sativa Seed Oil Effectively Replaces Fish Oil as a Dietary Source of Eicosapentaenoic Acid in Mice.
Fish currently supplies only 40% of the eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) required to allow all individuals globally to meet the minimum intake recommendation of 500 mg/d. Therefore, alternative sustainable sources are needed. The main objective was to investigate the ability of genetically engineered Camelina sativa (20% EPA) oil (CO) to enrich tissue EPA and DHA relative to an EPA-rich fish oil (FO) in mammals. Six-week-old male C57BL/6J mice were fed for 10 wk either a palm oil-containing control (C) diet or diets supplemented with EPA-CO or FO, with the C, low-EPA CO (COL), high-EPA CO (COH), low-EPA FO (FOL), and high-EPA FO (FOH) diets providing 0, 0.4, 3.4, 0.3, and 2.9 g EPA/kg diet, respectively. Liver, muscle, and brain were collected for fatty acid analysis, and blood glucose and serum lipids were quantified. The expression of selected hepatic genes involved in EPA and DHA biosynthesis and in modulating their cellular impact was determined. The oils were well tolerated, with significantly greater weight gain in the COH and FOH groups relative to the C group (P < 0.001). Significantly lower (36-38%) blood glucose concentrations were evident in the FOH and COH mice relative to C mice (P < 0.01). Hepatic EPA concentrations were higher in all EPA groups relative to the C group (P < 0.001), with concentrations of 0.0, 0.4, 2.9, 0.2, and 3.6 g/100 g liver total lipids in the C, COL, COH, FOL, and FOH groups, respectively. Comparable dose-independent enrichments of liver DHA were observed in mice fed CO and FO diets (P < 0.001). Relative to the C group, lower fatty acid desaturase 1 (Fads1) expression (P < 0.005) was observed in the COH and FOH groups. Higher fatty acid desaturase 2 (Fads2), peroxisome proliferator-activated receptor α (Ppara), and peroxisome proliferator-activated receptor γ (Pparg) (P < 0.005) expressions were induced by CO. No impact of treatment on liver X receptor α (Lxra) or sterol regulatory element-binding protein 1c (Srebp1c) was evident. Oil from transgenic Camelina is a bioavailable source of EPA in mice. These data provide support for the future assessment of this oil in a human feeding trial.


Ubiquitin-Specific Protease 8 Mutant Corticotrope Adenomas Present Unique Secretory and Molecular Features and Shed Light on the Role of Ubiquitylation on ACTH Processing.
Somatic mutations in the ubiquitin-specific protease 8 (USP8) gene have recently been shown to occur in ACTH-secreting pituitary adenomas, thus calling attention to the ubiquitin system in corticotrope adenomas. Assess the consequences of USP8 mutations and establish the role of ubiquitin on ACTH turnover in human ACTH-secreting pituitary adenomas. USP8 mutation status was established in 126 ACTH-secreting adenomas. Differences in ACTH secretion and POMC expression from adenoma primary cultures and in microarray gene expression profiles from archival specimens were sought according to USP8 sequence. Ubiquitin/ACTH coimmunoprecipitation and incubation with MG132, a proteasome inhibitor, were performed in order to establish whether ubiquitin plays a role in POMC/ACTH degradation in corticotrope adenomas. USP8 mutations were identified in 29 adenomas (23%). Adenomas presenting USP8 mutations secreted greater amounts of ACTH and expressed POMC at higher levels compared to USP wild-type specimens. USP8 mutant adenomas were also more sensitive to modulation by CRH and dexamethasone in vitro. At microarray analysis, genes associated with endosomal protein degradation and membrane components were downregulated in USP8 mutant adenomas as were AVPR1B, IL11RA, and PITX2. Inhibition of the ubiquitin-proteasome pathway increased ACTH secretion and POMC itself proved a target of ubiquitylation, independently of USP8 sequence status. Our study has shown that USP8 mutant ACTH-secreting adenomas present a more "typical" corticotrope phenotype and reduced expression of several genes associated with protein degradation. Further, ubiquitylation is directly involved in intracellular ACTH turnover, suggesting that the ubiquitin-proteasome system may represent a target for treatment of human ACTH-secreting adenomas.


Effect of propranolol on myocardial-infarct size in a randomized blinded multicenter trial.
A multicenter randomized single-blind study was performed to evaluate the effects of propranolol administered during the evolution of myocardial infarction. Five centers enrolled a total of 269 patients, with 134 receiving propranolol and 135 placebo. Propranolol or placebo was given intravenously upon randomization (0.1 mg per kilogram of body weight) and then orally for nine days to keep the heart rate between 45 and 60 beats per minute. Less than 2 per cent of patients were treated within 4 hours after the onset of symptoms, but 50 per cent received therapy within 8 hours of onset of chest pain, and the remainder between 8 and 18 hours. The heart rates in the propranolol-treated group were significantly lower than those in the placebo group (P less than 0.001). Base-line characteristics, including the mean heart rate (79.6 vs. 81.3) and the left ventricular ejection fraction (49.0 vs. 49.5), were similar in the two groups. The primary end point evaluated--infarct size as estimated from plasma MB creatine kinase activity--was virtually identical in the two groups, averaging 13.3 and 13.6 gram-equivalents of MB creatine kinase per square meter of body-surface area. Peak plasma levels of the enzyme were also similar in the two groups. No significant difference was observed between the propranolol and placebo groups in the change in left ventricular ejection fraction, extent of area involved in pyrophosphate uptake, R-wave loss on electrocardiograms, or mortality (after three years). These results do not support the use of propranolol administered four or more hours after the onset of symptoms to limit infarct size.


Sorghum stay-green QTL individually reduce post-flowering drought-induced leaf senescence.
Sorghum is an important source of food, feed, and biofuel, especially in the semi-arid tropics because this cereal is well adapted to harsh, drought-prone environments. Post-flowering drought adaptation in sorghum is associated with the stay-green phenotype. Alleles that contribute to this complex trait have been mapped to four major QTL, Stg1-Stg4, using a population derived from BTx642 and RTx7000. Near-isogenic RTx7000 lines containing BTx642 DNA spanning one or more of the four stay-green QTL were constructed. The size and location of BTx642 DNA regions in each RTx7000 NIL were analysed using 62 DNA markers spanning the four stay-green QTL. RTx7000 NILs were identified that contained BTx642 DNA completely or partially spanning Stg1, Stg2, Stg3, or Stg4. NILs were also identified that contained sub-portions of each QTL and various combinations of the four major stay-green QTL. Physiological analysis of four RTx7000 NILs containing only Stg1, Stg2, Stg3, or Stg4 showed that BTx642 alleles in each of these loci could contribute to the stay-green phenotype. RTx7000 NILs containing BTx642 DNA corresponding to Stg2 retained more green leaf area at maturity under terminal drought conditions than RTx7000 or the other RTx7000 NILs. Under post-anthesis water deficit, a trend for delayed onset of leaf senescence compared with RTx7000 was also exhibited by the Stg2, Stg3, and Stg4 NILs, while significantly lower rates of leaf senescence in relation to RTx7000 were displayed by all of the Stg NILs to varying degrees, but particularly by the Stg2 NIL. Greener leaves at anthesis relative to RTx7000, indicated by higher SPAD values, were exhibited by the Stg1 and Stg4 NILs. The RTx7000 NILs created in this study provide the starting point for in-depth analysis of stay-green physiology, interaction among stay-green QTL and map-based cloning of the genes that underlie this trait.


Childhood violence and mental health among indigenous Sami and non-Sami populations in Norway: a SAMINOR 2 questionnaire study.
The main objectives of this study were to investigate the association between childhood violence and psychological distress and post-traumatic stress symptoms (PTS) among Sami and non-Sami adults, and to explore a possible mediating effect of childhood violence on any ethnic differences in mental health. This study is part of a larger questionnaire survey on health and living conditions in Mid- and Northern Norway (SAMINOR 2) which included 2116 Sami and 8674 non-Sami participants. A positive association between childhood violence and psychological distress and PTS in adulthood was found regardless of ethnicity. For women, childhood violence may have mediated some of the ethnic differences in psychological distress (53.2%) and PTS (31.4%). A similar pattern was found for men as to psychological distress (45.5%) and PTS (55.5%). The prevalence of psychological distress was significantly higher in the Sami than in the non-Sami group: 15.8% vs. 13.0% for women, and 11.4% vs. 8.0% for men. Likewise, PTS showed a higher prevalence in the Sami group, both for women (16.2% vs. 12.4%) and for men (12.2% vs. 9.1). A positive association between childhood violence and adult mental distress was found for both Sami and Norwegian adults. More mental problems were found among the Sami. Childhood violence may have mediated some of the ethnic differences.


Characterization of measles virus-specific cytotoxic antibodies by use of a chronically infected cell line.
The complement-dependent cytotoxic effects of measles virus specific antibodies on a chronically infected cell line were studied by a microcytotoxicity assay. Cell damage was determined by trypan blue staining. Monospecific antisera against the hemagglutinin (HA) and the hemolysin (HL) of the virus were both cytotoxic. Rabbits, immunized with measles virus, developed cytotoxic antibodies in parallel with the appearance of antibodies against the measles virus HA and HL. Human sera with high antibody titers against HA and HL also gave a specific cytotoxic reaction. The main part of the activity was carried by IgG in rabbit sera tested within 2 weeks after primary immunization and only IgM. The serum reaction was characterized by a prozone. The maximal degree of cytotoxicity varied between tests performed on different occasions. Complement, at higher concentrations than were used for analysis of antibody-mediated cytolysis, caused a significant lysis of infected cells. Cerebrospinal fluid (CSF) samples containing measles virus-specific antibodies were investigated. CSF from patients with subacute sclerosing panencephalitis displayed a pronounced cytotoxicity without a prozone. CSF samples from four patients with multiple sclerosis were identified by a specific cytotoxic reaction despite low titers of antibodies against HA and HL.


Parameter interval estimation of system reliability for repairable multistate series-parallel system with fuzzy data.
The purpose of this paper is to create an interval estimation of the fuzzy system reliability for the repairable multistate series-parallel system (RMSS). Two-sided fuzzy confidence interval for the fuzzy system reliability is constructed. The performance of fuzzy confidence interval is considered based on the coverage probability and the expected length. In order to obtain the fuzzy system reliability, the fuzzy sets theory is applied to the system reliability problem when dealing with uncertainties in the RMSS. The fuzzy number with a triangular membership function is used for constructing the fuzzy failure rate and the fuzzy repair rate in the fuzzy reliability for the RMSS. The result shows that the good interval estimator for the fuzzy confidence interval is the obtained coverage probabilities the expected confidence coefficient with the narrowest expected length. The model presented herein is an effective estimation method when the sample size is n ≥ 100. In addition, the optimal α -cut for the narrowest lower expected length and the narrowest upper expected length are considered.


Temporal SNR characteristics in segmented 3D-EPI at 7T.
Three-dimensional segmented echo planar imaging (3D-EPI) is a promising approach for high-resolution functional magnetic resonance imaging, as it provides an increased signal-to-noise ratio (SNR) at similar temporal resolution to traditional multislice 2D-EPI readouts. Recently, the 3D-EPI technique has become more frequently used and it is important to better understand its implications for fMRI. In this study, the temporal SNR characteristics of 3D-EPI with varying numbers of segments are studied. It is shown that, in humans, the temporal variance increases with the number of segments used to form the EPI acquisition and that for segmented acquisitions, the maximum available temporal SNR is reduced compared to single shot acquisitions. This reduction with increased segmentation is not found in phantom data and thus likely due to physiological processes. When operating in the thermal noise dominated regime, fMRI experiments with a motor task revealed that the 3D variant outperforms the 2D-EPI in terms of temporal SNR and sensitivity to detect activated brain regions. Thus, the theoretical SNR advantage of a segmented 3D-EPI sequence for fMRI only exists in a low SNR situation. However, other advantages of 3D-EPI, such as the application of parallel imaging techniques in two dimensions and the low specific absorption rate requirements, may encourage the use of the 3D-EPI sequence for fMRI in situations with higher SNR.


Can social firms contribute to alleviating the economic burden of psychiatric disabilities for the public healthcare system?
In a number of countries, unemployment rates for people with psychiatric disabilities are much higher than in the general population. On the one hand, the expenses for mental health reach 3.5% of the total public health and social services budget in Québec. On the other hand, social firms (SFs) receive government subsidies. The objective was to compare public healthcare expenses for people with psychiatric disabilities who work in SFs with those associated with people with a similar condition who are looking for a job in the competitive labour market. This study followed a retrospective comparative design and considered two groups, namely: 122 employees working in SFs and 64 individuals participating in a supported employment program as job-seekers. Two complementary datasets were used: a self-report questionnaire and public healthcare databases. The cost analysis was performed from the perspective of the public healthcare system and included outpatient visit fees to physicians, outpatient visits to health professionals other than physicians in public healthcare centres, inpatient expenses due to hospitalisations, emergency room visits and amounts reimbursed to patients for medication. Regression analyses using generalised linear models with a gamma distribution and log link were used. Our results revealed that when controlling for sociodemographic variables (gender, age, marital status, education, physical disability), global health (EuroQol EQ-5D-5L), the severity of psychiatric symptoms (18-item Brief Symptom Inventory) and self-declared primary mental health diagnosis, annual healthcare costs paid by the public insurance system were between $1,924 and $3,912 lower for people working in SFs than for the comparison group. An explanatory hypothesis is that working in SFs could act as a substitute for medical treatments such as outpatient visits and medication use. There might be a form of compensation between supporting SFs and financing the public healthcare system, which provides valuable insights for public decision-making.


The alliance formation puzzle in contests with capacity-constraints: A test using American football reception-coverage contest data.
We utilize a contest-theoretic model to demonstrate a version of the alliance formation puzzle that aligns with reception-coverage contests in American football. Namely, secondary defenders can opt for single-coverage-1 v 1 contest. Alternatively, they can choose to ally-form double-coverage or 2 v 1 contest with exogenous intra-alliance prize division-when defending a given receiver. In our theoretical treatment, we find that defenses have a lower equilibrium success rate in preventing the receiver from "getting open" under double-coverage than under single-coverage in the absence of capacity constraints. We also find that this success rate paradox is a necessary condition for the alliance formation puzzle. We then test the theoretical treatment by analyzing 8,508 plays of NCAA and NFL game data within a set of fixed effects, logistic regression models that control for receiver, level-of-play, and season-of-play. We find that equilibrium level of defensive success rises significantly and substantially (p-value < 0.01 and marginal effect of between 13 and 17 percentage points) when moving from single-coverage to double-coverage, ceteris paribus. There is strong evidence that the necessary condition for the alliance formation puzzle does not hold in this setting. We conclude that sufficiently-binding physiological and training-based capacity constraints eliminate the alliance formation puzzle in this setting, as was shown theoretically by Konrad and Kovenock (2009). This empirical result suggests that other contest settings that regularly feature alliance, such as liquidity-constrained conflict, may not be puzzling.


Influence of Toxoplasma Gondii Infection on Symptoms and Signs of Premenstrual Syndrome: A Cross-sectional Study.
Infection with Toxoplasma gondii in brain may cause some symptoms that resemble those in women with premenstrual syndrome. To determine the association of T. gondii infection with symptoms and signs of premenstrual syndrome, we examined 489 women aged 30-40 years old. Sera of participants were analyzed for the presence of anti-Toxoplasma IgG and IgM antibodies using enzyme-linked immunoassays (EIA) and T. gondii DNA by polymerase chain reaction (PCR). Anti-T. gondii IgG antibodies were found in 38 (7.8%) of the women studied. Anti-T. gondii IgM antibodies were found in 13 (34.2%) of the 38 IgG seropositive women. Logistic regression showed two variables associated with seropositivity to T. gondii: presence of diarrhea (odds ratio [OR] = 6.10; 95% confidence interval [CI]: 1.37-27.85; P = 0.01) and weight gain (OR = 2.89; 95% CI: 1.37-6.07; P = 0.005), and two variables associated with high (>150 IU/ml) levels of IgG against T. gondii: presence of diarrhea (OR = 7.40; 95% CI: 1.79-30.46; P = 0.006) and abdominal inflammation (OR = 3.38; 95% CI: 1.13-10.10; P = 0.02). Positivity to EIA IgG and PCR was positively associated with obesity and negatively associated with joint pain by bivariate analysis. Our study for the first time reveals a potential association of T. gondii infection with clinical manifestations of premenstrual syndrome.


Characterization of Na/Ca exchange in plasmalemmal vesicles from zona fasciculata cells of the bovine adrenal gland.
The presence of an Na/Ca exchange system in fasciculata cells of the bovine adrenal gland was tested using isolated plasmalemmal vesicles. In the presence of an outwardly Na(+) gradient, Ca(2+) uptake was about 2-fold higher than in K(+) condition. Li(+) did not substitute for Na(+) and 5 mM Ni(2+) inhibited Ca(2+) uptake. Ca(2+) efflux from Ca(2+)-loaded vesicles was Na(+)-stimulated and Ni(2+)-inhibited. The saturable part of Na(+)-dependent Ca(2+) uptake displayed Michaelis-Menten kinetics. The relationship of Na(+)-dependent Ca(2+) uptake versus intravesicular Na(+) concentration was sigmoid (apparent K(0.5) approximately 24 mM; Hill number approximately 3) and Na(+) acted on V(max) without significant effect on K(m). Na(+)-stimulated Ca(2+) uptake was temperature-dependent (apparent Q(10) approximately 2.2). The inhibition properties of several divalent cations (Cd(2+), Sr(2+), Ni(2+), Ba(2+), Mn(2+), Mg(2+)) were tested and were similar to those observed in kidney basolateral membrane. The above results indicate the presence of an Na/Ca exchanger located on plasma membrane of zona fasciculata cells of bovine adrenal gland. This exchanger displays similarities with that of renal basolateral cell membrane.


Effects of four antiviral substances on lethal vaccinia virus (IHD strain) respiratory infections in mice.
Intranasal infection of BALB/c mice with the IHD strain of vaccinia virus was found to cause pneumonia, profound weight loss and death. Cidofovir, hexadecyloxypropyl-cidofovir (HDP-CDV), the diacetate ester prodrug of 2-amino-7-[(1,3-dihydroxy-2-propoxy)methyl]purine (HOE961), and ribavirin were used to treat the infections starting 24h after virus exposure. Single intraperitoneal (i.p.) cidofovir treatments of 100 and 30 mg/kg led to 90-100% survival compared with no survivors in the placebo group, whereas a 10 mg/kg dose was ineffective. The 100 mg/kg treatment reduced lung and snout virus titres on day 3 of the infection by 20- and 8-fold, respectively. Mean arterial oxygen saturation levels in these two cidofovir treatment groups were significantly higher than placebo on days 4 through 6 of the infection, indicating an improvement in lung function. Effects of cidofovir on viral pathogenesis were studied on days 1, 3 and 5 of the infection, and demonstrated statistically significant reductions in lung consolidation scores, lung weights, lung virus titre and snout virus titres on days 3 and 5. Cidofovir treatment also reduced virus titres in other tissues and body fluid, including blood, brain, heart, liver, salivary gland and spleen. HDP-CDV was given by oral gavage at 100, 50 and 25mg/kg doses one time only, resulting in 80-100% survival. Lower daily oral doses of 10 and 5mg/kg per day given for 5 days protected only 30% of animals from death. Oral doses (100, 50 and 25 mg/kg per day) of HOE961 for 5 days protected all animals, whereas equivalent oral doses of ribavirin were completely ineffective. The rapidity of recovery from weight loss during the infection was a function of dose of compound administered. These data indicate the utility of parenteral cidofovir, oral HDP-CDV and oral HOE961 in treating severe respiratory infections caused by this virus.


Autonomous and non-autonomous differentiation of ectoderm in different sea urchin species.
During early embryogenesis, the highly regulative sea urchin embryo relies extensively on cell-cell interactions for cellular specification. Here, the role of cellular interactions in the temporal and spatial expression of markers for oral and aboral ectoderm in Strongylocentrotus purpuratus and Lytechinus pictus was investigated. When pairs of mesomeres or animal caps, which are fated to give rise to ectoderm, were isolated and cultured they developed into ciliated embryoids that were morphologically polarized. In animal explants from S. purpuratus, the aboral ectoderm-specific Spec1 gene was activated at the same time as in control embryos and at relatively high levels. The Spec1 protein was restricted to the squamous epithelial cells in the embryoids suggesting that an oral-aboral axis formed and aboral ectoderm differentiation occurred correctly. However, the Ecto V protein, a marker for oral ectoderm differentiation, was detected throughout the embryoid and no stomodeum or ciliary band formed. These results indicated that animal explants from S. purpuratus were autonomous in their ability to form an oral-aboral axis and to differentiate aboral ectoderm, but other aspects of ectoderm differentiation require interaction with vegetal blastomeres. In contrast to S. purpuratus, aboral ectoderm-specific genes were not expressed in animal explants from L. pictus even though the resulting embryoids were morphologically very similar to those of S. purpuratus. Recombination of the explants with vegetal blastomeres or exposure to the vegetalizing agent LiCl restored activity of aboral ectoderm-specific genes, suggesting the requirement of a vegetal induction for differentiation of aboral ectoderm cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Torulopsis glabrata vaginitis.
To study the sociodemographic risk factors and clinical features of Torulopsis glabrata vaginal infection. We evaluated the sociodemographic and clinical characteristics of 86 consecutive symptomatic women attending a vaginitis clinic and isolated T glabrata. Case patients were compared with a control group of 174 asymptomatic women with negative vaginal cultures and an additional group of 625 symptomatic women with vaginal cultures positive for Candida albicans. In addition, the sensitivity of the isolates to the more common antimycotic agents used was tested by the modified Kirby-Bauer method. Patients with T glabrata vaginal infection were from lower socioeconomic backgrounds and had less education. They were more likely to use vaginal tampons and to be seropositive for human immunodeficiency virus than were negative controls. Compared with C albicans infection, T glabrata was more frequent among women over 38 years of age and in those with less education and of lower social class. In logistic regression analysis, T glabrata was associated more frequently with recurrent vaginal candidiasis than was C albicans (odds ratio 2.46, 95% confidence interval 1.33-4.54; P = .004). Six of the 86 (7%) T glabrata isolates and none of the C albicans isolates (P < .001 by Fisher exact test) were resistant to the imidazole derivatives tested. Torulopsis glabrata was isolated in 10% of women with vulvovaginal candidiasis attending a vaginitis clinic. This infection was associated with recurrent vaginitis in almost one-third of case patients presenting with symptoms.


A computational study of the potential energy surface of peroxyformic acid dimers.
MP2 and M05-2x calculations with aug-cc-pVDZ basis sets were used to analyze intermolecular interactions in peroxyformic acid dimers. A total of 18 and 16 minima were located on the potential energy surface of HOOCHO dimer complexes at M05-2x and MP2 computational levels, respectively. The BSSE corrected interaction energies are in a range between 9 and 34 kJ mol(-1) at the MP2/aug-cc-pVDZ computational level. The atoms-in-molecules (AIM) theory was also applied to explain the nature of the complexes. The interaction energies have been partitioned with the natural energy decomposition analysis (NEDA) showing that the most important attractive term corresponds to the charge transfer.


Chemical reactivities of catalytic and noncatalytic zinc or cobalt atoms of horse liver alcohol dehydrogenase: differentiation by their thermodynamic and kinetic properties.
Horse liver alcohol dehydrogenase (EC 18.104.22.168) contains one catalytic and one noncatalytic pair of zinc atoms that can be replaced selectively with cobalt and/or 65zince. We have now prepared "hybird" metalloenzymes by specifically replacing one or both pairs of zinc atoms with 65zinc and/or cobalt. Their differential chemical reactivities serve to characterize the metal atoms at either site. The spectral and kinetic properties of the resultant 65zinc, cobalt, and hybrid enzymes, as well as those of their complexes with 1,10-phenanthroline, identify the metal atoms that are at the catalytic sites and differentiate them from those at the noncatalytic sites. All data are in complete agreement with the results of the x-ray crystal structure analysis. Remarkably, under the conditions used, chemical reactivity, as gauged by thermodynamic methods under equilibrium conditions, identifies the catalytic metal atoms as those which are reactive to 1,10-phenanthroline, while this reagnet does not affect the noncatalytic pair. Under dynamic conditions the kinetics of the metal-metal exchange reveals the converse to be true: the chemical reactivity of the noncatalytic atoms is much higher and, hence, they exchange more rapidly. The results are examined in terms of thermodynamic and kinetic properties of metal complex ions which serve as the basis of possible mechanisms underlying these observations.


Dynamic CTC phenotypes in metastatic prostate cancer models visualized using magnetic ranking cytometry.
Tumors can shed thousands of cells into the circulation daily. These circulating tumor cells (CTCs) are heterogeneous, and their phenotypes change dynamically. Real-time monitoring of CTC phenotypes is crucial to elucidate the role of CTCs in the metastatic cascade. Here, we monitor phenotypic changes in CTCs in mice xenografted with tumors with varying aggressiveness during cancer progression and a course of chemotherapy to study the metastatic potential of CTCs and changes in the properties of these cells in response to treatment. A new device that enables magnetic ranking cytometry (MagRC) is employed to profile the phenotypic properties of CTCs. Overall, CTCs from metastatic xenografts in mice display dynamic and heterogeneous profiles while non-metastatic models had static profiles. Decreased heterogeneity followed by a reduction in metastasis incidence was observed after a course of chemotherapy administered to highly metastatic xenografts. Phenotypic profiling of CTCs could be employed to monitor disease progression and predict therapeutic responses.


A role for E1B-AP5 in ATR signaling pathways during adenovirus infection.
E1B-55K-associated protein 5 (E1B-AP5) is a cellular, heterogeneous nuclear ribonucleoprotein that is targeted by adenovirus (Ad) E1B-55K during infection. The function of E1B-AP5 during infection, however, remains largely unknown. Given the role of E1B-55K targets in the DNA damage response, we examined whether E1B-AP5 function was integral to these pathways. Here, we show a novel role for E1B-AP5 as a key regulator of ATR signaling pathways activated during Ad infection. E1B-AP5 is recruited to viral replication centers during infection, where it colocalizes with ATR-interacting protein (ATRIP) and the ATR substrate replication protein A 32 (RPA32). Indeed, E1B-AP5 associates with ATRIP and RPA complex component RPA70 in both uninfected and Ad-infected cells. Additionally, glutathione S-transferase pull-downs show that E1B-AP5 associates with RPA components RPA70 and RPA32 directly in vitro. E1B-AP5 is required for the ATR-dependent phosphorylation of RPA32 during infection and contributes to the Ad-induced phosphorylation of Smc1 and H2AX. In this regard, it is interesting that Ad5 and Ad12 differentially promote the phosphorylation of RPA32, Rad9, and Smc1 during infection such that Ad12 promotes a significant phosphorylation of RPA32 and Rad9, whereas Ad5 only weakly promotes RPA32 phosphorylation and does not induce Rad9 phosphorylation. These data suggest that Ad5 and Ad12 have evolved different strategies to regulate DNA damage signaling pathways during infection in order to promote viral replication. Taken together, our results define a role for E1B-AP5 in ATR signaling pathways activated during infection. This might have broader implications for the regulation of ATR activity during cellular DNA replication or in response to DNA damage.


Future Lyme disease risk in the south-eastern United States based on projected land cover.
Lyme disease is the most significant vector-borne disease in the United States, and its southward advance over several decades has been quantified. Previous research has examined the potential role of climate change on the disease's expansion, but no studies have considered the role of future land cover upon its distribution. This research examines Lyme disease risk in the south-eastern U.S. based on projected land cover developed under four Intergovernmental Panel on Climate Change Scenarios: A1B, A2, B1, and B2. Land cover types and edge indices significantly associated with Lyme disease in Virginia were incorporated into a spatial Poisson regression model to quantify potential land cover suitability for Lyme disease in the south-eastern U.S. under each scenario. Our results indicate an intensification of potential land cover suitability for Lyme disease under the A scenarios and a decrease of potential land cover suitability under the B scenarios. The decrease under the B scenarios is a critical result, indicating that Lyme disease risk can be decreased by making different land cover choices. Additionally, health officials can focus efforts in projected high incidence areas.


The influence of depression on physician-patient interaction in primary care.
Depression is common but not well diagnosed in primary care medicine. This study examines the influence of depression and its recognition on the physician-patient encounter. A total of 508 new adult patients were assigned randomly to 105 primary care providers. Self-reported depression was determined by the Beck Depression Inventory (BDI) on entry to the study. Initial visits were videotaped and analyzed using the Davis Observation Code. Chart notes were reviewed for diagnosis of depression. Seventy-seven of the 508 study patients (15%) were identified as depressed in chart notes, while 130 patients (26%) had a BDI score > or = 9, indicating moderate to severe depression. Recognition of depression was associated with increased counseling, decreased time conducting physical examination, and an increase in overall visit length. Both elevated BDI scores and physician recognition of depression were associated with decreases in chatting. Failure to recognize depression was associated with increased time taking medical history. Results support the potential value of psychological screening instruments in primary care and provide information for training physicians in the recognition and management of depression. The content of office visits is different when patients are depressed or are diagnosed as depressed.


The acute effect of ingesting a quercetin-based supplement on exercise-induced inflammation and immune changes in runners.
This study tested the acute anti-inflammatory and immune-modulating influence of a quercetin-based supplement consumed by endurance athletes 15 min before an intense 2-hr run. In this randomized, crossover study, 20 runners (11 men, 9 women, age 38.4 ± 2.1 yr) completed two 2-hr treadmill runs at 70% VO(2max) (3 wk apart). Subjects ingested either 4 quercetin-based chews (Q-chew) or placebo chews (PL) 15 min before the runs. The 4 Q-chews provided 1,000 mg quercetin, 120 mg epigallocatechin 3-gallate, 400 mg isoquercetin, 400 mg each eicosapentaenoic acid and docosahexaenoic acid, 1,000 mg vitamin C, and 40 mg niacinamide. Subjects provided blood samples 30 min before, immediately after, and 1 hr postexercise and were analyzed for plasma quercetin, total blood leukocytes (WBC), C-reactive protein (CRP), 9 cytokines (IL-6, TNFα, GM-CSF, IFNγ, IL-1β, IL-2, IL-8, IL-10, and IL-12p70), granulocyte (GR) and monocyte (MO) phagocytosis (PHAG), and oxidative-burst activity (OBA). Plasma quercetin increased from 80.0 ± 26.0 μg/L to 6,337 ± 414 μg/L immediately postexercise and 4,324 ± 310 μg/L 1 hr postexercise after ingestion of Q-chews, compared with no change in PL (p < .001). Exercise caused significant increases in, CRP, GM-CSF, IL-10, IL-1β, IL-2, IL-6, IL-8, TNFα, GR-PHAG, and MO-PHAG and decreases in GR-OBA and MO-OBA, but no differences in the pattern of change were measured between Q-chew and PL trials. Acute ingestion of Q-chews 15 min before heavy exertion caused a strong increase in plasma quercetin levels but did not counter postexercise inflammation or immune changes relative to placebo.


New drugs on the Internet: the case of Camfetamine.
The number of new psychoactive substances (NPS) advertised for sale online is constantly increasing and it has become a phenomenon of global concern. Among NPS, Camfetamine has been rediscovered as recreational drug in 2011. Very little information is still available in the scientific literature on its nature and potential health risks. Data in scientific literature were integrated with a multilingual qualitative assessment of a range of online resources over the period of 32 months (May 2011-January 2014). N-Methyl-3-phenyl-norbornan-2-amine (Camfetamine) may act as an indirect dopaminergic agonist in the central nervous system and may have mild-moderate opioid activity too. There are no current epidemiological data about recreational use of Camfetamine; our research shows that it is indeed used especially by individuals with a history of recreational polydrug misuse. It facilitates mental alertness, induces relaxation, and, unlike many other stimulants, seems not to be associated with severe physical effects. Valid causes for concern issued in our research may be Camfetamine intravenous or intramuscular administration as well as its use in conjunction with other psychoactive substances. It is here highlighted that more large-scale studies need to be carried out to confirm and better describe both the extent of Camfetamine misuse and possible psychotropic/adverse effects.


Factors associated with compliance with the recommended frequency of postnatal care services in three rural districts of Tanzania.
High neonatal mortality persists in Tanzania. Rates of decline are slow, in part because postnatal care (PNC) services for addressing this problem remain severely underutilized. This study assesses factors associated with utilization of PNC among mothers in rural Tanzania. This study analyzed household survey data collected in 2011 to understand health service utilization patterns among women of reproductive age and children less than 5 years of age in the Rufiji, Kilombero, and Ulanga districts of Tanzania. A total of 889 mothers were eligible for the current analysis. Multinomial logistic regression was used to determine factors associated with the likelihood of mothers seeking the WHO recommended PNC visits. The percent of newborns and their mothers with full PNC was low (10.4%). Factors explaining PNC completion were district of residence, ethnic group, pregnancy wantedness, ANC attendance, place of delivery, and any incidence of newborn. Mothers of unwanted pregnancies were less likely to attend PNC services compared to mothers of wanted pregnancies [for at least two PNC: aRRR = 0.57, 95% CI 0.35-0.94]. Sick newborns were more likely to receive PNC than newborns who were not sick during the first month after childbirth [for at least two PNC, aRRR = 3.52, 95% CI 2.12-5.86]. Mothers who attended ANC services more frequently were more likely to receive PNC services compared to those who had attended fewer than 2 ANC services [for 1 PNC, aRRR = 1.89, 95% CI 1.23-2.90]. Mothers who delivered at a health facility were less likely to attend PNC services compared to mothers who delivered outside a facility [for at least 2 PNC: aRRR = 0.42, 95% CI 0.26-0.76]. Model with interactions between ANC attendance and place of delivery shown that only ANC attendance had a positive and statistically significant effect on PNC visit. To achieve the WHO recommended number of PNC in rural Tanzania, our findings suggest the need to provide PNC through the community-based primary health care. Efforts to improve coverage of PNC should include expanding health education and counseling during childbirth and neonatal period to more effectively advocate PNC for newborns perceived to be healthy.


Endometriosis-associated ovarian cancer: a review of pathogenesis.
Endometriosis is classically defined as the presence of endometrial glands and stroma outside of the endometrial lining and uterine musculature. With an estimated frequency of 5%-10% among women of reproductive age, endometriosis is a common gynecologic disorder. While in itself a benign lesion, endometriosis shares several characteristics with invasive cancer, has been shown to undergo malignant transformation, and has been associated with an increased risk of epithelial ovarian carcinoma (EOC). Numerous epidemiologic studies have shown an increased risk of EOC among women with endometriosis. This is particularly true for women with endometrioid and clear cell ovarian carcinoma. However, the carcinogenic pathways by which endometriosis associated ovarian carcinoma (EAOC) develops remain poorly understood. Current molecular studies have sought to link endometriosis with EAOC through pathways related to oxidative stress, inflammation and hyperestrogenism. In addition, numerous studies have sought to identify an intermediary lesion between endometriosis and EAOC that may allow for the identification of endometriosis at greatest risk for malignant transformation or for the prevention of malignant transformation of this common gynecologic disorder. The objective of the current article is to review the current data regarding the molecular events associated with EAOC development from endometriosis, with a primary focus on malignancies of the endometrioid and clear cell histologic sub-types.


Evaluation of factors causing delayed graft function in live related donor renal transplantation.
To determine the incidence and determinants of delayed graft function due to post-transplant acute tubular necrosis in live related donor renal transplantation. This is a retrospective study of 337 recipients of live related donor renal graft performed between 1986 and 2006. Of these recipients, 24 (7.1%) subjects developed delayed graft function with no evidence of acute rejection, cyclosporin toxicity, vascular catastrophe or obstructive cause and had evidence of acute tubular necrosis (ATN Group). These subjects were compared with recipients (n= 313, 92.9%) who had no clinical or biochemical evidence of ATN. Mean age, and gender distribution of recipients was similar in the two groups (ATN group 35.7 +/- 8.3, non-ATN group 34.3 +/- 7.5, P= 0.43). Gender distribution of the recipients (men 279, 89.1% vs. 21, 87.5%, P= 0.80) as well as donors (women 221, 70.6% vs. 18, 75.0%, P= 0.75) was also similar. In ATN group as compared with non-ATN group the donor age was significantly greater (56.6 +/- 8.3 vs. 46.6 +/- 11.2 years, P< 0.0001). There was marginal difference in pre-operative systolic BP (154.5 +/- 18.3 vs. 147.4 +/- 20.2 mm Hg, P= 0.077) and significant difference in diastolic BP (87.8 +/- 9.5 vs. 83.4 +/- 11.4 mmHg, P= 0.041). Incidence of multiple renal arteries was similar (16.7% vs. 7.3%, P= 0.22). The warm ischemia time was significantly greater in ATN group (33.3 +/- 6.2 min) as compared to non-ATN group (30.4 +/- 5.7 min, P= 0.042). Duration of hospital stay was more in ATN group (19.9 +/- 6.7 vs. 16.8 +/- 8.4 days, P= 0.04) but there was no difference in 1 year survival (284 subjects, 90.7% vs. 21 subjects, 87.5%, P= 0.873). This study shows that greater donor age, higher baseline diastolic BP and greater warm ischemia time are major determinants of delayed graft function due to acute tubular necrosis after related donor renal transplantation.


Reduced instantaneous center of rotation movement in patients with low back pain.
The instantaneous center of rotation (ICR) can be used to investigate movement coordination and control in patients with low back pain (LBP). Tracking of the ICR in LBP patients has not been systematically investigated. This study aimed to (1) determine the within-session measurement error of ICR parameters, and (2) characterize the change in ICR among three groups of participants (no history of LBP = HC; history of LBP = HLBP; and current LBP = LBP). Ninety-three participants (HC = 31; HLBP = 33; and LBP = 29) were recruited. Participants performed two sets of three repetitions of an active forward bend, while their lumbar and pelvic movements were recorded with an electromagnetic tracking system. Total ICR displacement and the radius of the bounding sphere containing the ICR were derived during the forward bending and the return to upright phases. Intra-class correlation coefficients (ICC3,3) and minimal detectable difference (MDD) were used to determine measurement error and interpret findings of the group analysis. One-way ANOVAs and post hoc Bonferroni comparisons were used to determine differences among groups. ICC3,3 demonstrated excellent within-session test-retest reliability of the ICR parameters (ICC3,3 = 0.86-0.97). The MDD values were 0.20-3.40 mm. Comparisons between the HC and LBP groups and between the HLBP and LBP groups showed significant differences (p < 0.05) for all ICR parameters, with medium effect sizes (0.51-0.66), except for total displacement during forward bending between the HC and LBP groups. Less ICR displacement and variability in patients with LBP may indicate coping strategies to stiffen the lumbar spine. This could result from patients with LBP adopting a strategy of increased muscle activation to provide spinal stability during functional tasks.


Autoimmunity in hereditary retinal degeneration. I. Basic studies.
One hundred and sixteen patients with retinitis pigmentosa (RP), 64 patients with other eye diseases, and 36 control subjects with no known eye disease were examined for antiretinal autoimmune activity. Sera were screened by indirect immunofluorescence on normal donor human eye sections to detect antibodies to human retinal antigens. Forty three of 116 RP patients (37%), 21 out of 64 non-RP patients with other eye diseases (33%), and 1 out of 42 controls (2%) had antibodies reacting with donor eye retinal antigens. Lymphocytes were tested by an in vitro transformation assay to detect cell mediated immunity to retinal antigens. Sixteen RP patients (19%), 11 non-RP patients (18%), and four controls (10%) showed lymphocyte sensitisation. Autoimmune responses were detected in many degenerative ocular disorders, but it is not known if they play a contributory pathogenic role.


Changes in holothurian coelomocyte populations following immune stimulation with different molecular patterns.
Echinoderms possess a variety of cells populating the coelomic fluid; these cells are responsible for mounting defense against foreign agents. In the sea cucumber Holothuria glaberrima, four different coelomocyte types were readily distinguished using morphological, histochemical and physiological (phagocytic activity) parameters: lymphocytes, phagocytes, spherulocytes and "giant" cells (listed in order of abundance). Monoclonal antibodies generated against sea cucumber tissues and one polyclonal against sea urchin mayor yolk protein (MYP) were also used to characterize these cell populations. The effects of several pathogen-associated molecular patterns (PAMPs): Lipopolysaccharides from Escherichia. coli (LPS), heat-killed Staphylococcus aureus (SA) and a synthetic dsRNA were studied on coelomocyte cell populations. PAMPs increased the phagocytic activity of the holothurian coelomocytes, and were able to induce selective immune responses in several of these populations, demonstrating the ability of the sea cucumber to respond to a different variety of immune challenges. Overall, these results show the variety of cells that populate the coelomic fluid of the holothurian and demonstrate their involvement in immune reactions. These animals represent an untapped resource for new findings into the evolution and development of the immune response not only in invertebrates but also in phylogenetically shared reactions with vertebrates.


[Study on the role of the tubule in renal vasoconstriction induced by cyclosporine].
Cyclosporine (CyA) has proved to induce cell apoptosis on cultured proximal tubule cells. However, there is no much data about the in vivo functional consequences of this injury or the long time observed CyA-induced renal vasoconstriction. In a swine model of subacute CyA nephrotoxicity (10 mg/ Kg. dx 15 days), we performed a right nephrectomy, followed by left renal artery, vein and ureter catheterisati8n. After inducing water diuresis, three clearance periods of 15 minutes were performed before and after a furosemide 1 mg/kg infusion. Plasma and urine electrolytes, blood gas, acid excretion, plasma renin activity and aldosterone concentration, GFR, RPF, RBF, intra-renal vascular resistances, glomerular filtration pressure, distal Cl- delivery, water clearance and TTKG were measured or estimated on 7 control and 7 treated animals. Right kidney was processed for NaKATPase activity and immunostaining. Treated animals presented detaching proximal cells, luminal blebbing and loss of tight junctions. Cortical but not medullar sodium pump was internalised and partially inactive. Treated animals showed much lower fractional excretions of Na+, with significantly higher distal fractional reabsorption of Cl. Distal shift in fluid load resulted in a significant rise in renal O2 consumption, and modifications in the global renal estequiometry of Na+ transport/O2 uptake. Several consequences followed this situation: preglomerular resistances increased 3 times with only minor changes in postglomerular resistances and renal blood and plasma flow were significantly reduced. Furosemide partially reversed these effects. A slight increase in fractional filtration prevented GFR differences to become statistically significant. subacute CyA treatment even al doses not modifying GFR, may cause proximal tubule Na+ transport impairment, resulting in increased rates of distal delivery and absorption of fluid load. Renal uptake of O2 may be increased and tubule glomerular feedback should be expected to be activated. Absence of changes of GFR with furosemide is an early sign of CyA renal damage.


Probiotics in the arabian gulf region.
Available reports on adherence to recommended guidelines for labeling of probiotic products are based on assessment of these products in developed countries. In the Arabian Gulf region, there is a paucity of data on the characterization of probiotic products and an absence of local guidelines for their labeling. This study, carried out in the United Arab Emirates (UAE), represents the first evaluation of probiotic products available in the Arabian Peninsula. Probiotic products were purchased over the counter from a variety of sources, including pharmacies, healthfood stores, and supermarkets across the UAE. All identified products were listed and information regarding type of product preparation and labeling information were recorded. A total of 37 probiotic products, 15 dairy-based and 22 non-dairy-based were identified. The dairy products comprised of 12 yogurts, two fermented milk products and one powdered baby formula. The majority of non-dairy products were in capsule form (n = 16). While all the non-dairy products gave information about the strain of probiotic microorganism and number present at time of manufacture, this information was provided for only one dairy-based product. Strains of Lactobacillus acidophilus were the most common probiotic organisms identified. However, one probiotic product listed Enterococcus faecalis (750 million viable bacteria per capsule) as a component. With the exception of one non-dairy-based product, all health-related claims were structure/function statements, according to the US Food and Drug Administration nomenclature. These findings indicate that a wide variety of probiotic products are available in the Arabian Gulf. Development of guidelines for labeling of these probiotic products and use of structure/function statements and health claims should be addressed.


First Report of Bidens mottle virus Causing Mosaic and Leaf Deformation in Garland Chrysanthemum and Lettuce in Taiwan.
Garland chrysanthemum (Chrysanthemum coronarium) and lettuce (Lactuca sativa) are winter vegetables in Taiwan. Turnip mosaic virus (TuMV) and Lettuce mosaic virus (LMV) were potyviruses commonly isolated from garland chrysanthemum and lettuce, respectively (1). Symptoms of mosaic and deformation in leaves and stunting of plants have been observed in both Compositae crops in the fields since 2007 in the Chiayi area and with an increasing incidence in recent years (26 and 33% in garland chrysanthemum and lettuce, respectively). Filamentous virus particles (approximately 780 × 13 nm) in the crude sap and pinwheel inclusions in infected cells can be observed in the preparations of both diseased hosts with electron microscopy. However, TuMV, LMV, and other potyviruses (Bean yellow mosaic virus, Papaya ringspot virus, and Zucchini yellow mosaic virus) were not detectable in diseased samples by ELISA and western blotting tests, indicating a new potyvirus infection. Virus cultures were isolated from infected garland chrysanthemum and lettuce separately via mechanical inoculations in Chenopodium quinoa. Each isolate was mechanically inoculated to their original host individually and all caused symptoms similar to that observed in the field, indicating their pathogenicity to their original host. A cDNA fragment consisting of partial nuclear inclusion (NIb) and coat protein (CP) genes were amplified with potyvirus degenerate primers (forward: 5'-GGBAAYAATAGTGGNCAACC and reverse: 5'-GGGGAGGTGCCGTTCTCDATRCACCA) and was found to share 90% nucleotide sequence identity to that of Bidens mottle virus (BiMoV; GenBank Accession No. AF538686). The sequences of the CP gene and 3' untranslated region (3'-UTR) of tested viruses were further amplified with a specific primer of BiMoV CP and oligo-dT in reverse transcription-PCR. The amplified fragments were cloned, sequenced, and the combined cDNA sequences were deposited in GenBank (Accession No. AB491763 for isolate garland chrysanthemum and Accession No. AB491764 for isolate lettuce). Sequence analysis showed that both cloned sequences shared more than 97% nucleotide similarity to that of BiMoV. The amino acid sequence of the CP of both isolates shared a 99.3% identity and a 98.9 to 99.3% identity to that of other BiMoV isolates deposited in GenBank. BiMoV was first described to be infecting lettuce and Cichorium endivia in the United States (4) and was first reported in sunflower and calendula in Taiwan recently (2,3). To our knowledge, this is the first report of the occurrence of BiMoV in garland chrysanthemum and lettuce in Taiwan. References: (1) Y. K. Chen et al. Plant Pathol. Bull. 5:55,1996. (2) C.-H. Huang and F.-J. Jan. Plant Dis. 95:362, 2011. (3) J. Y. Liao et al. Arch. Virol. 154:723, 2009. (4) F. Youssef et al. Arch. Virol 153:227, 2008.


Biochemical (prostate-specific antigen) relapse-free survival and toxicity after 125I low-dose-rate prostate brachytherapy.
To report our clinical experience and 5-year prostate-specific antigen (PSA) relapse-free survival rate for early-stage prostate cancer after (125)I low-dose-rate prostate brachytherapy. In all, 300 patients were treated between March 1999 and April 2003, and followed prospectively. Patients were stratified into low-, intermediate- and high-risk groups, and those receiving neoadjuvant androgen deprivation (NAAD) or not. Kaplan-Meier estimates of PSA relapse-free survival and PSA nadirs were obtained for all patients and for the risk groups. Toxicity, as urinary and erectile dysfunction (ED), were reported from a prospective database. The median (range) follow-up was 45 (33-82) months. The actuarial PSA relapse-free survival was 93% at 5 years; 21 (7%) of patients had evidence of biochemical failure as defined by the American Society of Therapeutic Radiation Oncology criteria. There was no significant difference in actuarial survival for patients in the different risk groups, or between those receiving NAAD or not (low-risk 96%, intermediate 89%, high 93%, P = 0.12; NAAD 92%, no NAAD 95%, P = 0.30). Overall the 3-year median PSA level was 0.3 ng/mL (192 men). There was no significant difference in median 3-year PSA levels for different risk groups, or for those treated with or with no NAAD. The 3- and 4-year PSA nadir of <0.5 ng/mL was achieved by 71% and 86% of men, respectively. The acute urinary retention rate was 7%; 5.6% of men developed urethral strictures requiring dilatation, while 2.7% required a transurethral resection of the prostate after implantation, for obstructive symptoms. Of patients with no ED before treatment, 62% had no ED at 2 years, and of these 60% used a phosphodiesterase inhibitor. This prospective series confirms the excellent overall biochemical survival after (125)I brachytherapy; the treatment was tolerated well, with early and late urinary toxicity and ED similar to other published results.


Genetic organization and sequence of the Pseudomonas cepacia genes for the alpha and beta subunits of protocatechuate 3,4-dioxygenase.
The locations of the genes for the alpha and beta subunits of protocatechuate 3,4-dioxygenase (EC 18.104.22.168) on a 9.5-kilobase-pair PstI fragment cloned from the Pseudomonas cepacia DBO1 chromosome were determined. This was accomplished through the construction of several subclones into the broad-host-range cloning vectors pRO2317, pRO2320, and pRO2321. The ability of each subclone to complement mutations in protocatechuate 3,4-dioxygenase (pcaA) was tested in mutant strains derived from P. cepacia, Pseudomonas aeruginosa, and Pseudomonas putida. These complementation studies also showed that the two subunits were expressed from the same promoter. The nucleotide sequence of the region encoding for protocatechuate 3,4-dioxygenase was determined. The deduced amino acid sequence matched that determined by N-terminal analysis of regions of the isolated enzyme. Although over 400 nucleotides were sequenced before the start of the genes, no homology to known promoters was found. However, a terminator stem-loop structure was found immediately after the genes. The deduced amino acid sequence showed extensive homology with the previously determined amino acid sequence of protocatechuate 3,4-dioxygenase from another Pseudomonas species.


Disparities in new graduate transition from multiple stakeholder perspectives.
The purpose of this qualitative study was to understand multiple stakeholder perspectives of new graduate (NG) transition programs. It was part of a larger mixed-methods study (2011) designed to provide a comprehensive assessment of new graduate nurse transition best practices, across six British Columbia health authorities. Data collection involved individual interviews with academic nurse educators (n=4) and separate focus groups with new graduate (n=48) and front-line nurse leaders (n=69). Disparity emerged as the overriding theme and described differences between stakeholder group perspectives, between expectations and reality, and within and across programs. Four disparities emerged: entry-level education and practice, perspectives on employment and career planning, transition program elements and support. Despite general satisfaction with undergraduate preparation, theory-practice gaps were identified. New Graduates experienced misalignments between their employment expectations and their realities. The employed student nurse program in which many new graduates had participated did not always yield employment, but when it did, differences in transitional expectations arose between new graduates and leaders. There was considerable variation across and within provincial new graduate programs with respect to orientation, supernumerary time and preceptorship characteristics, including lack of training. Disparities arose in the nature, amount of and access to support and the monitoring of new graduate progress. Findings reinforced organizational complexities and the importance of communication across education and practice sectors. This paper uncovers the tensions between the perspectives of new graduates and nurse leaders about transitional programs and opens the opportunity to collaborate in aligning the perspectives.


The outcome of peripheral T-cell lymphoma patients failing first-line therapy: a report from the prospective, International T-Cell Project.
This analysis explored factors influencing survival of patients with primary refractory and relapsed peripheral T-cell lymphomas enrolled in the prospective International T-cell Project. We analyzed data from 1020 patients with newly diagnosed disease, enrolled between September 2006 and December 2015. Out of 937 patients who received first-line treatment, 436 (47%) were identified as refractory and 197 (21%) as relapsed. Median time from the end of treatment to relapse was 8 months (range 2-73). Overall, 75 patients (8%) were consolidated with bone marrow transplantation, including 12 refractory and 22 relapsed patients. After a median follow up of 38 months (range 1-96 months) from documentation of refractory/relapsed disease, 440 patients had died. The median overall survival (OS) was 5.8 months; 3-year overall survival rates were 21% and 28% for refractory and relapsed patients, respectively (P<0.001). Patients receiving or not salvage bone marrow transplantation had a 3-year survival of 48% and 18%, respectively (P<0.001). In a univariate Cox regression analysis, refractory disease was associated with a higher risk of death (HR=1.43, P=0.001), whereas late relapse (>12 months, HR 0.57, P=0.001) and salvage therapy with transplantation (HR=0.36, P<0.001) were associated with a better OS. No difference was found in OS with respect to histology. This study accurately reflects outcomes for patients treated according to standards of care worldwide. Results confirm that peripheral T-cell lymphomas patients had dismal outcome after relapse or progression. Patients with chemotherapy sensitive disease who relapsed after more than 12 months might benefit from consolidation bone marrow transplantation.


Design and methodological considerations of an effectiveness trial of a computer-assisted intervention: an example from the NIDA Clinical Trials Network.
Computer-assisted interventions hold the promise of minimizing two problems that are ubiquitous in substance abuse treatment: the lack of ready access to treatment and the challenges to providing empirically-supported treatments. Reviews of research on computer-assisted treatments for mental health and substance abuse report promising findings, but study quality and methodological limitations remain an issue. In addition, relatively few computer-assisted treatments have been tested among illicit substance users. This manuscript describes the methodological considerations of a multi-site effectiveness trial conducted within the National Institute on Drug Abuse's (NIDA's) National Drug Abuse Treatment Clinical Trials Network (CTN). The study is evaluating a web-based version of the Community Reinforcement Approach, in addition to prize-based contingency management, among 500 participants enrolled in 10 outpatient substance abuse treatment programs. Several potential effectiveness trial designs were considered and the rationale for the choice of design in this study is described. The study uses a randomized controlled design (with independent treatment arm allocation), intention-to-treat primary outcome analysis, biological markers for the primary outcome of abstinence, long-term follow-up assessments, precise measurement of intervention dose, and a cost-effectiveness analysis. Input from community providers during protocol development highlighted potential concerns and helped to address issues of practicality and feasibility. Collaboration between providers and investigators supports the utility of infrastructures that enhance research partnerships to facilitate effectiveness trials and dissemination of promising, technologically innovative treatments. Outcomes from this study will further the empirical knowledge base on the effectiveness and cost-effectiveness of computer-assisted treatment in clinical treatment settings.


Genetic parameters estimates for ewes' behavioural reactivity towards their litter after lambing.
In livestock, improving maternal reactivity towards the litter is an important issue in breeding strategies to promote production and animal welfare. As of yet, no studies have investigated the within-breed genetic variation of maternal reactivity in sheep. The objective of this study was to estimate the genetic parameters of maternal reactivity traits. A total of 1,095 primiparous and 1,441 multiparous Romane ewes were phenotyped 24 hr postlambing using a behavioural test (arena test, AT) over a 10-year experimental period. The test consisted of three successive phases evaluating the ewe's attraction to her litter, reactivity to separation from her litter, and reactivity to a conflict between attraction to her litter and avoidance of a motionless human. The ewes were reared exclusively on rangelands (South of France) and lambed outdoors in the spring. High-pitched bleating and low-pitched bleating in the AT were mostly highly heritable (0.39-0.46). Heritabilities were moderate for proximity to the litter in the presence of a human (0.27) and low for locomotion and vigilance in the AT (0.09-0.15). The measurements of a given behaviour in the three phases of the AT were highly genetically correlated. Few genetic correlations were found between the different behavioural traits in the AT, the highest correlations being between high-pitched bleating and low-pitched bleating (-0.43 to -0.77). In conclusion, our findings demonstrate moderate-to-high heritability for maternal reactivity traits. These traits could be included in genetic selection schemes to enhance maternal attachment provided there is no unfavourable link with other production traits.


Self-propulsion of a catalytically active particle near a planar wall: from reflection to sliding and hovering.
Micron-sized particles moving through a solution in response to self-generated chemical gradients serve as model systems for studying active matter. Their far-reaching potential applications will require the particles to sense and respond to their local environment in a robust manner. The self-generated hydrodynamic and chemical fields, which induce particle motion, probe and are modified by that very environment, including confining boundaries. Focusing on a catalytically active Janus particle as a paradigmatic example, we predict that near a hard planar wall such a particle exhibits several scenarios of motion: reflection from the wall, motion at a steady-state orientation and height above the wall, or motionless, steady "hovering." Concerning the steady states, the height and the orientation are determined both by the proportion of catalyst coverage and the interactions of the solutes with the different "faces" of the particle. Accordingly, we propose that a desired behavior can be selected by tuning these parameters via a judicious design of the particle surface chemistry.