Study protocol for statin web-based investigation of side effects (StatinWISE): a series of randomised controlled N-of-1 trials comparing atorvastatin and placebo in UK primary care.
Statins are effective at preventing cardiovascular disease, widely prescribed and their use is growing. Uncertainty persists about whether they cause symptomatic muscle adverse effects, such as pain and weakness, in the absence of statin myopathy. Discrepancies between data from observational studies, which suggest statins are associated with excess muscle symptoms, and from randomised trials, which suggest no such excess, have caused confusion. N-of-1 trials offer the opportunity to establish whether muscle symptoms during statin use are caused by statins in particular individuals. This series of 200 randomised, double-blinded N-of-1 trials in primary care will determine (1) the effect of statins on all muscle symptoms and (2) the effect of statins on muscle pain that is perceived to be statin related. Patients who are considering discontinuing statin use due to muscle symptoms and those who have discontinued in the last 3 years due to such symptoms will be recruited. Participants will be randomised to a sequence of six 2-month treatment periods during which they will receive atorvastatin 20 mg per day or matched placebo. On each of the last 7 days of each treatment period, participants will rate their muscle symptoms on a Visual Analogue Scale (VAS).At the end of their trial, participants will be shown numerical and graphical summaries of their own symptom data during statin and placebo periods. The primary analysis on the aggregate data from all participants will be a linear mixed model for VAS muscle symptom score, comparing scores during treatment with statin and placebo. This trial received a favourable opinion from South Central-Hampshire A Research Ethics Committee. Results will be published in a peer-reviewed medical journal. Dissemination of results to patients will take place via the media, website (statinwise.lshtm.ac.uk) and patient organisations. ISRCTN30952488.


Adverse events in women and children who have received intrapartum antibiotic prophylaxis treatment: a systematic review.
Adverse events from intrapartum antibiotic prophylaxis (IAP) are poorly documented yet essential to inform clinical practice for neonatal group B Streptococcus (GBS) disease prevention. In this systematic review, we appraised and synthesised the evidence on the adverse events of IAP in the mother and/or her child. We searched MEDLINE, MEDLINE In-Process & Other Non-Indexed Citations, EMBASE, Cochrane, and Science Citation Index from date of inception until October 16th 2016. Reference lists of included studies and relevant systematic reviews were hand-searched. We included primary studies in English that reported any adverse events from intrapartum antibiotics for any prophylactic purpose compared to controls. The search was not restricted to prophylaxis for GBS but excluded women with symptoms of infection or undergoing caesarean section. Two reviewers assessed the methodological quality of studies, using the Cochrane Risk of Bias tool, and the Risk of Bias Assessment Tool for Nonrandomised Studies. Results were synthesised narratively and displayed in text and tables. From 2364 unique records, 30 studies were included. Despite a wide range of adverse events reported in 17 observational studies and 13 randomised controlled trials (RCTs), the evidence was inconsistent and at high risk of bias. Only one RCT investigated the long-term effects of IAP reporting potentially serious outcomes such as cerebral palsy; however, it had limited applicability and unclear biological plausibility. Seven observational studies showed that IAP for maternal GBS colonisation alters the infant microbiome. However, study populations were not followed through to clinical outcomes, therefore clinical significance is unknown. There was also observational evidence for increased antimicrobial resistance, however studies were at high or unclear risk of bias. The evidence base to determine the frequency of adverse events from intrapartum antibiotic prophylaxis for neonatal GBS disease prevention is limited. As RCTs may not be possible, large, better quality, and longitudinal observational studies across countries with widespread IAP could fill this gap. CRD42016037195 .


Populations and determinants of airborne fungi in large office buildings.
Bioaerosol concentrations in office environments and their roles in causing building-related symptoms have drawn much attention in recent years. Most bioaerosol studies have been cross-sectional. We conducted a longitudinal study to examine the characteristics of airborne fungal populations and correlations with other environmental parameters in office environments. We investigated four office buildings in Boston, Massachusetts, during 1 year beginning May 1997, recruiting 21 offices with open workstations. We conducted intensive bioaerosol sampling every 6 weeks resulting in 10 sets of measurement events at each workstation, and recorded relative humidity, temperature, and CO2 concentrations continuously. We used principal component analysis (PCA) to identify groups of culturable fungal taxa that covaried in air. Four major groupings (PCA factors) were derived where the fungal taxa in the same groupings shared similar ecological requirements. Total airborne fungal concentrations varied significantly by season (highest in summer, lowest in winter) and were positively correlated with relative humidity and negatively related to CO2 concentrations. The first and second PCA factors had similar correlations with environmental variables compared with total fungi. The results of this study provide essential information on the variability within airborne fungal populations in office environments over time. These data also provide background against which cross-sectional data can be compared to facilitate interpretation. More studies are needed to correlate airborne fungi and occupants' health, controlling for seasonal effects and other important environmental factors.


Use of placebo controls in the evaluation of surgery: systematic review.
To investigate whether placebo controls should be used in the evaluation of surgical interventions. Systematic review. We searched Medline, Embase, and the Cochrane Controlled Trials Register from their inception to November 2013. Randomised clinical trials comparing any surgical intervention with placebo. Surgery was defined as any procedure that both changes the anatomy and requires a skin incision or use of endoscopic techniques. Three reviewers (KW, BJFD, IR) independently identified the relevant trials and extracted data on study details, outcomes, and harms from included studies. In 39 out of 53 (74%) trials there was improvement in the placebo arm and in 27 (51%) trials the effect of placebo did not differ from that of surgery. In 26 (49%) trials, surgery was superior to placebo but the magnitude of the effect of the surgical intervention over that of the placebo was generally small. Serious adverse events were reported in the placebo arm in 18 trials (34%) and in the surgical arm in 22 trials (41.5%); in four trials authors did not specify in which arm the events occurred. However, in many studies adverse events were unrelated to the intervention or associated with the severity of the condition. The existing placebo controlled trials investigated only less invasive procedures that did not involve laparotomy, thoracotomy, craniotomy, or extensive tissue dissection. Placebo controlled trial is a powerful, feasible way of showing the efficacy of surgical procedures. The risks of adverse effects associated with the placebo are small. In half of the studies, the results provide evidence against continued use of the investigated surgical procedures. Without well designed placebo controlled trials of surgery, ineffective treatment may continue unchallenged.


Control of intestinal inflammation by interleukin-10.
Twenty years ago, the observation that mice genetically deficient in IL-10 spontaneously developed severe intestinal inflammation, revealed an essential role for IL-10 in the maintenance of intestinal homeostasis. In the intervening period much has been learned about the cellular and molecular factors that are involved in IL-10-mediated regulatory pathways. Elegant experiments with conditional cell-type specific knockout strains have illustrated that IL-10 acts on both myeloid cells and T cells within the intestine to suppress innate and adaptive inflammatory responses and enhance regulatory circuits. Although several distinct cellular sources of IL-10 have been identified in the gut, CD4(+) T cells are a crucial non-redundant source of IL-10 for the regulation of intestinal inflammation. Induction of IL-10 may represent an important means through which intestinal microbiota establishes mutually beneficial commensalism with mammalian hosts, but can be exploited by certain pathogens to facilitate infection. Recent genetic studies in humans have confirmed the essential role of IL-10 in preventing deleterious inflammation in the gut. A better understanding of the molecular pathways involved in IL-10 induction and function in the intestine may facilitate the development of novel therapies for inflammatory bowel disease (IBD).


Sonographically abnormal placenta: an association with an increased risk poor pregnancy outcomes.
The placenta is a most interesting but unfortunately often ignored and misunderstood organ. Placental abnormalities, therefore, can be an "early warning system" for fetal problems. A complete prenatal sonographic examination of the placenta is an essential component as its abnormalities can have a direct effect on fetal or maternal outcomes, obstetrical management and future fertility. To determine whether any association exists between the finding of an increased thickness of placenta, abnormal placenta shape, placental calcification, placental lake and abnormal cord insertion site at 20-22 and 30-32 weeks gestation with an increased risk of uteroplacental complications or a poor pregnancy outcome. A real-time ultrasound was used at the time of detail scan (at 20-22 weeks gestation) and at 30-32 weeks gestation to look for placenta appearance, fetal growth and anomaly. The main outcome measures were risk of hypertension disease in pregnancy, fetal growth restriction and poor fetal outcomes such as low Apgar score and low cord pH. The majority of the participants were Malay (77.9%). Abnormal placenta found at both gestations were placental lakes and thickness, and only one case had marginal cord insertion. Approximately 6% of the cases were confirmed placenta previa. No abnormal shape or abnormal calcification found at both gestations. About 10% patient developed hypertensive disease in pregnancy, 15% of the fetus was found to have growth restriction and another 16% have low umbilical cord pH. Majority of them delivered at term (90%) and via vaginal delivery (81%). There was no significance between presence of abnormal placental lake and thickness at both gestations with the maternal and fetal outcome. Presence of abnormal placental thickness and lakes at 30-32 weeks scan associated with maternal hypertensive disease, fetal growth restriction and low umbilical cord pH, however these were not statistically significant.


Radical Surgery in Advanced Cervical Cancer Patients Receiving Bevacizumab-Containing Chemotherapy: A "Real Life Experience".
Platinum-based chemotherapy plus bevacizumab is the new standard of care in stage IVB cervical cancer (CC) patients. In this new scenario, radical surgery could be offered in selected cases with an optimal clinical response. Potential surgical complications related to previous bevacizumab treatment have never been described before in this type of setting. This is a single-institutional retrospective study on women with CC who received radical pelvic surgery after platinum-based chemotherapy (platinum-taxol) plus bevacizumab. Between April 2016 and September 2017, 15 patients with CC underwent radical surgery after bevacizumab-containing chemotherapy (platinum-taxol) at the Division of Gynecologic Oncology, Catholic University of the Sacred Heart in Rome, Italy.All patients received at least 3 cycles of treatment. Fourteen (93.4%) patients underwent radical hysterectomy, whereas 1 patient received pelvic anterior exenteration. Median operating time was 290 minutes (range, 220-400), and estimated blood loss was 250 mL (range, 50-1500). Median time of hospitalization was 5 days (range, 2-21). Pathological response in the cervix was complete in 3 cases and microscopic in 1 patient (26.7%). Intraoperative complications occurred in 3 cases: 2 vascular injuries and 1 ureteral injury. Three patients (20.0% of the whole series) experienced grade 3 complications.At last follow-up (median time, 9 months), the median overall survival is 13 months (range, 6-31): 7 (46.7%) women were free of disease, 8 (53.3%) patients progressed, and 3 of them died of disease. Radical surgery can be performed in women with CC previously treated with a bevacizumab-containing regimen.


Kupffer cell cytotoxicity to hepatocytes in coculture requires L-arginine.
Activated macrophages convert L-arginine to citrulline and unstable nitrogen oxides that have cytotoxic properties. We recently have shown that the inhibition of protein synthesis in Kupffer cell (KC):hepatocyte (HC) coculture, following exposure to gram-negative bacterial endotoxin (lipopolysaccharide), is due to the metabolism of L-arginine by this cytotoxic pathway. Although this finding supports a role for activated KCs and the L-arginine-dependent mechanism in the HC dysfunction seen in sepsis, it and previous studies have failed to demonstrate direct damage to HCs by adjacent KCs. The current study was undertaken to determine if KCs exposed to lipopolysaccharide could directly damage HCs and, if so, whether the damage was dependent on the metabolism of L-arginine. By using the release of aspartate aminotransferase as a marker of HC damage, it was found that a significant aspartate aminotransferase release by KC:HC cocultures in response to lipopolysaccharide occurred only if L-arginine was present. In addition, requirements for significant aspartate aminotransferase release included KC:HC ratios of 7.5:1 or greater and L-arginine concentrations of 1 mmol or more. Although the KC-induced damage was mild, these results show that in vitro HC damage in KC:HC coculture does require the metabolism of L-arginine and supports a hypothesis that toxic L-arginine metabolites may contribute to liver cell damage in patients with sepsis.


Cough and sleep in inner-city children.
This study aimed to determine whether cough at night keeps children awake, to describe the relationship between children's cough and sleep and to report parents' perceptions of their children's cough and sleep. Thirty-nine children with reported persistent cough at night (>3 weeks) were recruited and studied for 6 nights by video-recording. Coughs were counted and sleep state was coded for awake, restless sleep and quiet sleep. The relationships between cough and sleep state between subjects and within subjects were examined by correlation and regression. After night 2 the parents were asked whether their child had coughed or had disturbed sleep and after night 6 they were asked whether there had been any change. There was a weak relationship between log percentage of the night awake and log number of coughs (r=0.13, SE 0.036), and log (percentage of the night awake plus restless sleep) and log number of coughs (r=0.016, SE 0.0071). If the relationship between cough and sleep state is causal, halving the number of coughs will reduce the percentage of the night awake by 9% (95% confidence interval (CI) 4-15%) and percentage awake and restless by 1% (95% CI 0-2%). All but one parent correctly identified coughing and 82% detected change. Most could not comment on their child's sleep. Improvement in cough would result in little reduction in either the percentage of the night awake or awake and restless in the average child in the population studied. Parents could detect whether their children were coughing but not whether their sleep was disturbed.


Fatal child abuse in Georgia: the epidemiology of severe physical child abuse.
Decisions about the occurrence of child abuse are increasingly difficult to make because concepts of what qualifies as reportable child abuse may be broadening. We examined this question by comparing 51 fatal child abuse cases occurring in Georgia between July 1975 and December 1979 to non-fatal cases and to the Georgia population. Overall rates of fatal child abuse were higher for male perpetrators compared with female and black perpetrators compared with white. However, the latter finding varied with economic and geographic status. The highest child abuse fatality rates were found in poor, rural, white families (3.3/100,000 children) and in poor, urban, black families (2.4/100,000 children). Risk factors for fatal abuse included early childhood (RR 6:1), parental teenage childbearing (RR 4:1), and low socioeconomic status. These characteristics were similar to those of the severe child abuse cases noted in the early child abuse literature. Non-fatal cases did not clearly share these risk factors. Severe abuse, here represented by fatal cases, is a distinct subset of reported child abuse, but characteristics associated with it are frequently attributed to all reportable child abuse. Medical personnel should be aware that they cannot rely on the presence or absence of these characteristics in screening for risk of reportable child abuse. Child abuse research should use restricted, stated case definitions. When intervention and prevention programs are being organized, they should not generalize research findings to all forms of child abuse.


Quantitation of ER Structure and Function.
The plant endoplasmic reticulum forms a network of tubules connected by three-way junctions or sheet-like cisternae. Although the network is three-dimensional, in many plant cells, it is constrained to a thin volume sandwiched between the vacuole and plasma membrane, effectively restricting it to a 2-D planar network. The structure of the network, and the morphology of the tubules and cisternae can be automatically extracted following intensity-independent edge-enhancement and various segmentation techniques to give an initial pixel-based skeleton, which is then converted to a graph representation. Collectively, this approach yields a wealth of quantitative metrics for ER structure and can be used to describe the effects of pharmacological treatments or genetic manipulation. The software is publicly available.


De novo mutations in synaptic transmission genes including DNM1 cause epileptic encephalopathies.
Emerging evidence indicates that epileptic encephalopathies are genetically highly heterogeneous, underscoring the need for large cohorts of well-characterized individuals to further define the genetic landscape. Through a collaboration between two consortia (EuroEPINOMICS and Epi4K/EPGP), we analyzed exome-sequencing data of 356 trios with the "classical" epileptic encephalopathies, infantile spasms and Lennox Gastaut syndrome, including 264 trios previously analyzed by the Epi4K/EPGP consortium. In this expanded cohort, we find 429 de novo mutations, including de novo mutations in DNM1 in five individuals and de novo mutations in GABBR2, FASN, and RYR3 in two individuals each. Unlike previous studies, this cohort is sufficiently large to show a significant excess of de novo mutations in epileptic encephalopathy probands compared to the general population using a likelihood analysis (p = 8.2 × 10(-4)), supporting a prominent role for de novo mutations in epileptic encephalopathies. We bring statistical evidence that mutations in DNM1 cause epileptic encephalopathy, find suggestive evidence for a role of three additional genes, and show that at least 12% of analyzed individuals have an identifiable causal de novo mutation. Strikingly, 75% of mutations in these probands are predicted to disrupt a protein involved in regulating synaptic transmission, and there is a significant enrichment of de novo mutations in genes in this pathway in the entire cohort as well. These findings emphasize an important role for synaptic dysregulation in epileptic encephalopathies, above and beyond that caused by ion channel dysfunction.


Evaluation of two formulated chitin synthesis inhibitors, hexaflumuron and lufenuron against the raisin moth, Ephestia figulilella.
The raisin moth, Ephestia figulilella Gregson (Lepidoptera: Pyralidae), has a nearly cosmopolitan distribution, and causes severe quantitative and qualitative losses throughout the world. The larvae attack various drying and dried fruits, fallen figs, and damaged or moldy clusters of grapes on vines. Control of this pest in storage depends mostly on synthetic pesticides with several adverse side effects. To mitigate the adverse effects of these pesticides, investigations have focused on the development of compounds with more selectivity, and short residual life. In this research, insecticidal effects of two chitin synthesis inhibitors, hexaflumuron and lufenuron, were investigated against E. figulilella. Graded concentrations of each pesticide were prepared with distilled water. One-day-old fifth instar were sprayed by Potter's precision spray tower. Application of hexaflumuron and lufenuron on last instar larvae of E. figulilella caused not only mortality in larval stage, but also caused defects in pupal and adult stages. Larval mortality increased as concentration increased. The longevity of the fifth instars in both hexaflumuron and lufenuron treatments, in comparison with the controls, increased by more than 12 days. The longevity of adults decreased by about 10 days. Probit analysis data revealed that the sensitivity of the test insect to hexaflumuron (EC(50) = 95.38 ppm) was greater than lufenuron (EC(50)= 379.21 ppm).


Accelerated corneal graft rejection in baby rats.
Penetrating keratoplasty in infants has a very poor outcome compared to adults. It is of intrinsic interest to gain insight into the still unknown immunological mechanisms of graft failure because any form of uncorrected corneal opacity leads to amblyopia. Allogeneic keratoplasty was performed between Lewis and Fisher rats. The recipients' ages were 10 and 3 weeks, respectively. All experiments were controlled syngeneically. Survival rates were calculated and cellular infiltrates analysed histologically. Median graft survival times were 15 days in old recipients and 9 days in young recipients (p<0.01). There were fewer infiltrating cells in the younger rats than in the older ones on the day of rejection. Despite the fact that T cells dominated there were significantly more NK cells in young recipients at all time points after transplantation when compared to old recipients. An animal model has been established that shows similar rejection kinetics as in children, that is corneal graft failure occurs sooner in young rats. Already little infiltration was sufficient to reject a corneal allograft. The dominance of infiltrating NK cells and the vigorous rejection process suggest an involvement of the innate immune system in this process.


Effect of pectin feeding on obesity development and duodenal alkaline phosphatase activity in Sprague-Dawley rats fed with high-fat/high-energy diet.
Purpose The objective of this study was to evaluate whether pectin feeding would affect the small intestinal function and whether these changes would lead to obesity prevention in rats fed with high-fat diet. Three groups of weaned male rats (ad lib. fed; rats fed with diet containing 15% w/w of citrus pectin; restrictedly pair-fed rats) were fed with either a standard diet (9.5% fat) or a high-fat diet (30% fat) for 10 days. Results Our results revealed that pectin feeding led to significant decreases in body weight, energy intake and fat pad weight in rats fed with the standard as well as high-fat diet. Moreover, compared to the restrictedly pair-fed rats, in both groups of rats fed with the diet containing pectin, significant decrease in duodenal alkaline phosphatase (AP) activity was observed in histochemically stained cryostat sections. In contrast, despite their lower energy intake, restrictedly pair-fed rats showed similar fat pad deposition accompanied by unchanged values of AP activity in comparison to the controls. Conclusions Our findings indicate that daily pectin consumption could be beneficial in suppressing body weight gain and reducing probability of obesity risk in rats fed with a high-fat diet.


Impact of a posttraumatic cerebral infarction on outcome in patients with TBI: the Italian multicenter cohort INCEPT study.
Post-traumatic cerebral infarction (PTCI) is common after traumatic brain injury (TBI). It is unclear what the occurrence of a PTCI is, how it impacts the long-term outcome, and whether it adds incremental prognostic value to established outcome predictors. This was a prospective multicenter cohort study of moderate and severe TBI patients. The primary objective was to evaluate if PTCI was an independent risk factor for the 6-month outcome assessed with the Glasgow Outcome Scale (GOS). We also assessed the PTCI occurrence and if it adds incremental value to the International Mission for Prognosis and Clinical Trial design in TBI (IMPACT) core and extended models. We enrolled 143 patients, of whom 47 (32.9%) developed a PTCI. In the multiple ordered logistic regression, PTCI was retained in both the core and extended IMPACT models as an independent predictor of the GOS. The predictive performances increased significantly when PTCI was added to the IMPACT core model (AUC = 0.73, 95% C.I. 0.66-0.82; increased to AUC = 0.79, 95% CI 0.71-0.83, p = 0.0007) and extended model (AUC = 0.74, 95% C.I. 0.65-0.81 increased to AUC = 0.80, 95% C.I. 0.69-0.85; p = 0.00008). Patients with PTCI showed higher ICU mortality and 6-month mortality, whereas hospital mortality did not differ between the two groups. PTCI is a common complication in patients suffering from a moderate or severe TBI and is an independent risk factor for long-term disability. The addition of PTCI to the IMPACT core and extended predictive models significantly increased their performance in predicting the GOS. The present study was registered in ClinicalTrial.gov with the ID number NCT02430324.


Novel metabolic attributes of the genus cyanothece, comprising a group of unicellular nitrogen-fixing Cyanothece.
The genus Cyanothece comprises unicellular cyanobacteria that are morphologically diverse and ecologically versatile. Studies over the last decade have established members of this genus to be important components of the marine ecosystem, contributing significantly to the nitrogen and carbon cycle. System-level studies of Cyanothece sp. ATCC 51142, a prototypic member of this group, revealed many interesting metabolic attributes. To identify the metabolic traits that define this class of cyanobacteria, five additional Cyanothece strains were sequenced to completion. The presence of a large, contiguous nitrogenase gene cluster and the ability to carry out aerobic nitrogen fixation distinguish Cyanothece as a genus of unicellular, aerobic nitrogen-fixing cyanobacteria. Cyanothece cells can create an anoxic intracellular environment at night, allowing oxygen-sensitive processes to take place in these oxygenic organisms. Large carbohydrate reserves accumulate in the cells during the day, ensuring sufficient energy for the processes that require the anoxic phase of the cells. Our study indicates that this genus maintains a plastic genome, incorporating new metabolic capabilities while simultaneously retaining archaic metabolic traits, a unique combination which provides the flexibility to adapt to various ecological and environmental conditions. Rearrangement of the nitrogenase cluster in Cyanothece sp. strain 7425 and the concomitant loss of its aerobic nitrogen-fixing ability suggest that a similar mechanism might have been at play in cyanobacterial strains that eventually lost their nitrogen-fixing ability. The unicellular cyanobacterial genus Cyanothece has significant roles in the nitrogen cycle in aquatic and terrestrial environments. Cyanothece sp. ATCC 51142 was extensively studied over the last decade and has emerged as an important model photosynthetic microbe for bioenergy production. To expand our understanding of the distinctive metabolic capabilities of this cyanobacterial group, we analyzed the genome sequences of five additional Cyanothece strains from different geographical habitats, exhibiting diverse morphological and physiological attributes. These strains exhibit high rates of N(2) fixation and H(2) production under aerobic conditions. They can generate copious amounts of carbohydrates that are stored in large starch-like granules and facilitate energy-intensive processes during the dark, anoxic phase of the cells. The genomes of some Cyanothece strains are quite unique in that there are linear elements in addition to a large circular chromosome. Our study provides novel insights into the metabolism of this class of unicellular nitrogen-fixing cyanobacteria.


Facile assembly for fast construction of intercalation hybrids of layered double hydroxides with anionic metalloporphyrin.
Anionic manganese tetrasulfonatophenyl porphyrin (MnTSPP) has been intercalated into the interlamellar space of Mg-Al and Ni-Al layered double hydroxides (LDHs) through the exfoliation/restacking approach by using exfoliated LDH nanosheets and guest molecules as building blocks. The obtained hybrids were characterized by a variety of analytical techniques such as CHN analysis, XRD, FTIR, SEM, HRTEM, UV-vis spectroscopy and thermal analysis. Interlayer spacings determined from XRD patterns reveal a perpendicular orientation of the MnTSPP anions between the hyroxylated layers of both LDHs. The results of zeta potential measurements give information about the surface charge change of LDH nanoparticles associated with the spontaneous coassembly process. The catalytic performance of the heterogeneous catalysts MnTSPP/Mg-Al LDH2.0 and MnTSPP/Ni-Al LDH1.0 for the epoxidation of cyclohexene was investigated using molecular oxygen as an oxidant and isobutylaldehyde as a co-reductant. The intercalated hybrids appear to be promising catalysts owing to their good catalytic activity and selectivity.


Sertraline in children and adolescents with obsessive-compulsive disorder: a multicenter randomized controlled trial.
The serotonin reuptake inhibitors are the treatment of choice for patients with obsessive-compulsive disorder; however, empirical support for this assertion has been weaker for children and adolescents than for adults. To evaluate the safety and efficacy of the selective serotonin reuptake inhibitor sertraline hydrochloride in children and adolescents with obsessive-compulsive disorder. Randomized, double-blind, placebo-controlled trial. One hundred eighty-seven patients: 107 children aged 6 to 12 years and 80 adolescents aged 13 to 17 years randomized to receive either sertraline (53 children, 39 adolescents) or placebo (54 children, 41 adolescents). Twelve US academic and community clinics with experience conducting randomized controlled trials. Sertraline hydrochloride was titrated to a maximum of 200 mg/d during the first 4 weeks of double-blind therapy, after which patients continued to receive this dosage of medication for 8 more weeks. Control patients received placebo. The Children's Yale-Brown Obsessive Compulsive Scale (CY-BOCS), the National Institute of Mental Health Global Obsessive Compulsive Scale (NIMH GOCS), and the NIMH Clinical Global Impressions of Severity of Illness (CGI-S) and Improvement (CGI-I) rating scales. In intent-to-treat analyses, patients treated with sertraline showed significantly greater improvement than did placebo-treated patients on the CY-BOCS (adjusted mean, -6.8vs -3.4, respectively; P=.005), the NIMH GOCS (-2.2 vs -1.3, respectively; P=.02), and the CGI-I (2.7 vs 3.3, respectively; P=.002) scales. Significant differences in efficacy between sertraline and placebo emerged at week 3 and persisted for the duration of the study. Based on CGI-I ratings at end point, 42% of patients receiving sertraline and 26% of patients receiving placebo were very much or much improved. Neither age nor sex predicted response to treatment. The incidence of insomnia, nausea, agitation, and tremor were significantly greater in patients receiving sertraline; 12 (13%) of 92 sertraline-treated patients and 3 (3.2%) of 95 placebo-treated patients discontinued prematurely because of adverse medical events (P=.02). No clinically meaningful abnormalities were apparent on vital sign determinations, laboratory findings, or electrocardiographic measurements. Sertraline appears to be a safe and effective short-term treatment for children and adolescents with obsessive-compulsive disorder.


Epstein-Barr virus-associated peripheral T-cell lymphoma and hemophagocytic syndrome arising after liver transplantation: case report and review of the literature.
Post-transplantation lymphoproliferative disorders (PTLD) are a well-recognized complication of solid organ transplantation. The vast majority of PTLD are Epstein-Barr virus (EBV)-related infections that manifest as B-cell malignancies. We report an unusual case of an EBV-associated T-cell lymphoma in a 10-year-old boy who had previously undergone liver transplantation at age 4 years. He presented with hemophagocytic syndrome (HPS) and active EBV infection, with positive serum titers and polymerase chain reaction (PCR) for EBV in blood, colon, and antral samples.


Interventions in hospitalised patients with cancer: the importance of impending death awareness.
Burdensome and futile interventions with the aim of prolonging life should be avoided in dying patients. However, current clinical practice has hardly been investigated.We examined the number and type of diagnostic and therapeutic medical interventions in hospitalised patients with cancer in their last days of life. In addition, we investigated if physician awareness of impending death affected the use of these interventions. Questionnaire study and medical record study. Attending physicians of patients who died in a university hospital between January 2010 and June 2012 were asked whether they had been aware of the patient's impending death. The use of diagnostic and therapeutic interventions and medications was assessed by studying patients' charts. We included 131 patients. In the last 72 and 24 hours of life, 59% and 24% of the patients received one or more diagnostic interventions, respectively. Therapeutic interventions were provided to 47% and 31%. In the last 24 hours of life, patients received on average 5.8 types of medication.Awareness of a patient's impending death was associated with a significant lower use of diagnostic interventions (48% vs 69% in the last 72 hours; 11% vs 37% in the last 24 hours) and several medications that potentially prolong life (eg, antibiotics and cardiovascular medication). Many patients with cancer who die in hospital receive diagnostic and therapeutic interventions in the last days of life of which their advantages are questionable. To improve end-of-life care, medical care should be adapted.


In vivo and in vitro glucuronidation of the flavonoid diosmetin in rats.
Flavonoids form an important family of compounds widely present in plants and therefore in food, sometimes as substitutes for synthetic antioxidants. Although the excretion routes of flavonoids in animals has been explored, little is known about the details of their conjugation in the xenobiotic metabolism pathway. In this study, we investigate the metabolism of diosmetin as a model compound in the rat, particularly its level in blood after treatment (100 mg/kg, po) and the presence of its glucuronide(s) in both blood and urine. We demonstrate that after po treatment of the rat, a rapid glucuronidation takes place and that diosmetin circulates as glucuronides, whereas no free diosmetin is present in blood and urine. The glucuronides formed are present in the blood plasma at a high level (approximately 10 micrograms/ml), for at least 6 hr after the treatment and the conjugates are excreted in urine. We have detected four different glucuronides in blood and characterized the two major ones after purification by a combination of MS, NMR, and UV spectroscopy: diosmetin-7,3'-diglucuronide and diosmetin-3'-glucuronide. A brief characterization of the in vitro glucuronidation of some flavonoid compounds using liver microsomal preparations suggests that this important class of natural compounds might be conjugated by a specific isoform of UDP-glucuronosyltransferase. Thus, this work brings evidence that diosmetin is rapidly glucuronoconjugated in the rat and provides an explanation for the low po bioavailability of the unchanged compound that can be generalized to this important class of natural compounds.


Situ: Identifying and Explaining Suspicious Behavior in Networks.
Despite the best efforts of cyber security analysts, networked computing assets are routinely compromised, resulting in the loss of intellectual property, the disclosure of state secrets, and major financial damages. Anomaly detection methods are beneficial for detecting new types of attacks and abnormal network activity, but such algorithms can be difficult to understand and trust. Network operators and cyber analysts need fast and scalable tools to help identify suspicious behavior that bypasses automated security systems, but operators do not want another automated tool with algorithms they do not trust. Experts need tools to augment their own domain expertise and to provide a contextual understanding of suspicious behavior to help them make decisions. In this paper we present Situ, a visual analytics system for discovering suspicious behavior in streaming network data. Situ provides a scalable solution that combines anomaly detection with information visualization. The system's visualizations enable operators to identify and investigate the most anomalous events and IP addresses, and the tool provides context to help operators understand why they are anomalous. Finally, operators need tools that can be integrated into their workflow and with their existing tools. This paper describes the Situ platform and its deployment in an operational network setting. We discuss how operators are currently using the tool in a large organization's security operations center and present the results of expert reviews with professionals.


Meta-analysis of mismatch repair polymorphisms within the cogent consortium for colorectal cancer susceptibility.
In the last four years, Genome-Wide Association Studies (GWAS) have identified sixteen low-penetrance polymorphisms on fourteen different loci associated with colorectal cancer (CRC). Due to the low risks conferred by known common variants, most of the 35% broad-sense heritability estimated by twin studies remains unexplained. Recently our group performed a case-control study for eight Single Nucleotide Polymorphisms (SNPs) in 4 CRC genes. The present investigation is a follow-up of that study. We have genotyped six SNPs that showed a positive association and carried out a meta-analysis based on eight additional studies comprising in total more than 8000 cases and 6000 controls. The estimated recessive odds ratio for one of the SNPs, rs3219489 (MUTYH Q338H), decreased from 1.52 in the original Swedish study, to 1.18 in the Swedish replication, and to 1.08 in the initial meta-analysis. Since the corresponding summary probability value was 0.06, we decided to retrieve additional information for this polymorphism. The incorporation of six further studies resulted in around 13000 cases and 13000 controls. The newly updated OR was 1.03. The results from the present large, multicenter study illustrate the possibility of decreasing effect sizes with increasing samples sizes. Phenotypic heterogeneity, differential environmental exposures, and population specific linkage disequilibrium patterns may explain the observed difference of genetic effects between Sweden and the other investigated cohorts.


The perceived impact of location privacy: a web-based survey of public health perspectives and requirements in the UK and Canada.
The "place-consciousness" of public health professionals is on the rise as spatial analyses and Geographic Information Systems (GIS) are rapidly becoming key components of their toolbox. However, "place" is most useful at its most precise, granular scale - which increases identification risks, thereby clashing with privacy issues. This paper describes the views and requirements of public health professionals in Canada and the UK on privacy issues and spatial data, as collected through a web-based survey. Perceptions on the impact of privacy were collected through a web-based survey administered between November 2006 and January 2007. The survey targeted government, non-government and academic GIS labs and research groups involved in public health, as well as public health units (Canada), ministries, and observatories (UK). Potential participants were invited to participate through personally addressed, standardised emails. Of 112 invitees in Canada and 75 in the UK, 66 and 28 participated in the survey, respectively. The completion proportion for Canada was 91%, and 86% for the UK. No response differences were observed between the two countries. Ninety three percent of participants indicated a requirement for personally identifiable data (PID) in their public health activities, including geographic information. Privacy was identified as an obstacle to public health practice by 71% of respondents. The overall self-rated median score for knowledge of privacy legislation and policies was 7 out of 10. Those who rated their knowledge of privacy as high (at the median or above) also rated it significantly more severe as an obstacle to research (P < 0.001). The most critical cause cited by participants in both countries was bureaucracy. The clash between PID requirements - including granular geography - and limitations imposed by privacy and its associated bureaucracy require immediate attention and solutions, particularly given the increasing utilisation of GIS in public health. Solutions include harmonization of privacy legislation with public health requirements, bureaucratic simplification, increased multidisciplinary discourse, education, and development of toolsets, algorithms and guidelines for using and reporting on disaggregate data.


The clinical and immunological features of pediatric COVID-19 patients in China.
In December 2019, the corona virus disease 2019 (COVID-19) caused by novel coronavirus (SARS-CoV-2) emerged in Wuhan, China and rapidly spread worldwide. Few information on clinical features and immunological profile of COVID-19 in paediatrics. The clinical features and treatment outcomes of twelve paediatric patients confirmed as COVID-19 were analyzed. The immunological features of children patients was investigated and compared with twenty adult patients. The median age was 14.5-years (range from 0.64 to 17), and six of the patients were male. The average incubation period was 8 days. Clinically, cough (9/12, 75%) and fever (7/12, 58.3%) were the most common symptoms. Four patients (33.3%) had diarrhea during the disease. As to the immune profile, children had higher amount of total T cell, CD8+ T cell and B cell but lower CRP levels than adults (P < 0.05). Ground-glass opacity (GGO) and local patchy shadowing were the typical radiological findings on chest CT scan. All patients received antiviral and symptomatic treatment and the symptom relieved in 3-4 days after admitted to hospital. The paediatric patients showed mild symptom but with longer incubation period. Children infected with SARS-CoV-2 had different immune profile with higher T cell amount and low inflammatory factors level, which might ascribed to the mild clinical symptom. We advise that nucleic acid test or examination of serum IgM/IgG antibodies against SARS-CoV-2 should be taken for children with exposure history regardless of clinical symptom.


Ectopia lentis et pupillae in four generations caused by novel mutations in the ADAMTSL4 gene.
To identify the phenotype, genetic defect and inheritance pattern of ectopia lentis et pupillae (ELP) in a large Dutch family, previously diagnosed as presumed autosomal dominant ELP because of the occurrence of ELP in three generations. A clinical and genetic study of children and adults. Eight patients of the ELP family, including five new patients from the youngest generation. Standard ophthalmological examinations were performed. For molecular genetic analysis, the coding region of ADAMTSL4 was sequenced. Main outcome measures were the ocular phenotype of the new ELP patients, the inheritance pattern and the identification of mutations in the ADAMTSL4 gene in the family. Of the eight patients with ectopia lentis, seven fulfilled the clinical diagnostic criteria of ELP. Molecular genetic analysis of these seven patients disclosed two novel mutations in the ADAMTSL4 gene: homozygous (p.Q752X/p.Q752X) in six patients and compound heterozygous (p.Q752X/p.Q758fs) in one patient. Heterozygosity in phenotypically normal parents proved autosomal recessive (AR) inheritance. The pseudodominant inheritance pattern can be explained by high carrier frequency in this small community and/or consanguinity. Patients from a family with ELP in four generations have AR ELP caused by novel mutations in ADAMTSL4. The clinical presentation of ELP can be variable, but all patients of our study with homozygous p.Q752X mutation have ectopia lentis and pupillary dysfunction in common.


The role of B7 costimulation by murine acute myeloid leukemia in the generation and function of a CD8+ T-cell line with potent in vivo graft-versus-leukemia properties.
Relapse is more frequent after autologous than allogeneic bone marrow transplantation (BMT), due in part to lack of T-lymphocyte mediated allogeneic graft-versus-leukemia (GVL) effects. Infusions of leukemia-reactive T cells to patients after autologous BMT may be a means for providing a GVL effect. Costimulation of T cells by binding of the CD28 receptor on T cells with B7-counter receptors on antigen presenting cells amplifies antigen-specific T-cell responses. To enhance generation of leukemia reactive cytotoxic T lymphocytes (CTL), the murine B7-1- and B7-2-costimulatory molecule cDNAs were introduced into the MHC class I+, class II-, murine meyloid leukemia cell line C1498. B7-1 expression greatly enhanced the ability of the leukemia cells to generate and expand leukemia reactive CTL in vitro. A highly cytolytic and C1498 specific CD8+ CTL line was generated by B7-1 costimulation. This CTL line proliferated autonomously and produced interleukin-2 when provided B7-1 or B7-2 costimulation by C1498 leukemia cells. To test the in vivo antileukemia properties of this CTL line, irradiated syngeneic BMT recipients were given graded doses of leukemia cells on day 0, followed by CTL infusions beginning on day 1 post-BMT. Recipients of 10(7) CTL had a 3 log reduction in leukemia burden such that 100% of mice were protected from a supralethal leukemic cell dose. Sustained immune responses were detectable up to 3 months postinfusion of the CTL line. B7-1 or B7-2 costimulation in vivo did not augment antileukemia effects of infused CTL post BMT. These results suggest that B7 costimulation of leukemia reactive CTL may be important for their ex vivo generation and expansion for use in human adoptive immunotherapy of leukemia.


Intradermal skin test results correlate with atopy.
Intradermal skin testing (IDST) is performed by most allergists, but its value remains controversial. In most previous studies assessing the value of IDST, a positive result was based on a 5- to 6-mm wheal and erythema. While a subject's immediate reaction upon allergen exposure is a common endpoint to determine "allergy," additional known allergic phenomena such as the delayed or late-phase response to allergen exposure suggest that correlation only with current history or acute challenge may result in an incorrect labeling of false-positive results when, in fact, there is a physiologic response to exposure, albeit a reaction not immediately evident. We conducted a retrospective study to determine if positive IDST reactions represent nonspecific irritation or if they correlate with atopy. We retrospectively reviewed the records of 100 patients who had undergone skin prick testing (SPT) and IDST and compiled data on their age, sex, diagnosis, and number of skin and intradermal reactions. Results were analyzed according to a general linear model to see if the development of a positive IDST result correlated with atopy (defined as SPT positivity). We found statistically significant differences between SPT-positive and -negative patients with regard to diagnoses of asthma, rhinitis, or both (p=0.008). Controlling for asthma and rhinitis, we also found a significant association between atopy and a positive IDST result; among atopic (SPT+) patients, a mean of 25.9% of IDST results were positive, compared with a rate of only 6.7% IDST positivity among the nonatopic (SPT-) patients (p<0.0001). We conclude that IDST is more commonly positive in atopic (SPT+) than nonatopic (SPT-) patients, suggesting that a positive IDST represents genuine atopy and bona fide sensitization rather than nonspecific irritation.


Blood-stage challenge for malaria vaccine efficacy trials: a pilot study with discussion of safety and potential value.
There is increasing interest in malaria vaccines targeting the asexual blood stage of Plasmodium falciparum. Without accepted immunologic correlates of clinical protection, challenge studies are useful for assessing the efficacy of candidate vaccines in vivo in healthy volunteers. We report a pilot study of a safe and robust challenge protocol using a blood-stage inoculum. We have applied well-validated trial endpoints and twice daily real-time quantitative polymerase chain reaction monitoring of parasitemia to blood-stage challenge, which enabled direct comparison with sporozoite challenge. We found that greater accuracy in quantification of blood-stage growth rates can be achieved with blood-stage challenge. This finding may provide greater power to detect partial efficacy of many blood-stage candidate vaccines. We discuss the potential utility of blood-stage challenge studies in accelerating malaria vaccine development.


Knowledge, attitudes and factors associated with acceptability of human papillomavirus vaccination among undergraduate medical, dental and nursing students in South India.
Purpose: To evaluate knowledge, attitudes and factors associated with acceptability of human papillomavirus (HPV) vaccine among undergraduate medical, dental, and nursing students in South India. Methods: Using a post-test study design, a convenience sample of 988 students (age 18-26 years) were surveyed. The primary outcome was vaccine acceptability (likely to intend to receive the vaccine). Bivariate analysis using Chi-Square test of independence and multivariate binary logistic regression analysis was used to examine factors associated with vaccine acceptability. Results: Out of 988 students surveyed, majority had heard about cervical cancer (95%), HPV (89.3) or genital warts(77.5). Only 59.7% had heard of HPV vaccine prior to the survey; 65.2% intended to receive the vaccine and 68.3% were willing to recommend the vaccine to others. Participants aged <22 years were less likely to accept the vaccine (OR:0.85, CI:0.76-0.96) compared with participants aged older than 22 years. Medical students (OR:1.12, CI:1.03-1.23), students who reported alcohol use (OR:1.15, CI:1.03-1.29) and those with moderate knowledge scores were more likely to intend to receive the vaccine (OR:1.14, CI:1.04-1.24), compared to others. On multivariate analysis, only course (OR 1.366, CI 1.016-1.835) and attitude score (OR 4.17; CI 2.12-8.2) were statistically associated with intention to receive the HPV vaccine. Conclusion: Two-thirds of students intended to receive the HPV vaccine. Although the overall awareness of the HPV-related disease and prevention is good, considerable knowledge gaps exists in many areas suggesting that that more education about HPV disease and benefits of vaccination should be included in the undergraduate medical school curriculum.


Personalized oncology through integrative high-throughput sequencing: a pilot study.
Individual cancers harbor a set of genetic aberrations that can be informative for identifying rational therapies currently available or in clinical trials. We implemented a pilot study to explore the practical challenges of applying high-throughput sequencing in clinical oncology. We enrolled patients with advanced or refractory cancer who were eligible for clinical trials. For each patient, we performed whole-genome sequencing of the tumor, targeted whole-exome sequencing of tumor and normal DNA, and transcriptome sequencing (RNA-Seq) of the tumor to identify potentially informative mutations in a clinically relevant time frame of 3 to 4 weeks. With this approach, we detected several classes of cancer mutations including structural rearrangements, copy number alterations, point mutations, and gene expression alterations. A multidisciplinary Sequencing Tumor Board (STB) deliberated on the clinical interpretation of the sequencing results obtained. We tested our sequencing strategy on human prostate cancer xenografts. Next, we enrolled two patients into the clinical protocol and were able to review the results at our STB within 24 days of biopsy. The first patient had metastatic colorectal cancer in which we identified somatic point mutations in NRAS, TP53, AURKA, FAS, and MYH11, plus amplification and overexpression of cyclin-dependent kinase 8 (CDK8). The second patient had malignant melanoma, in which we identified a somatic point mutation in HRAS and a structural rearrangement affecting CDKN2C. The STB identified the CDK8 amplification and Ras mutation as providing a rationale for clinical trials with CDK inhibitors or MEK (mitogen-activated or extracellular signal-regulated protein kinase kinase) and PI3K (phosphatidylinositol 3-kinase) inhibitors, respectively. Integrative high-throughput sequencing of patients with advanced cancer generates a comprehensive, individual mutational landscape to facilitate biomarker-driven clinical trials in oncology.


Increased tetanic force and reduced myoplasmic [P(i)] following a brief series of tetani in mouse soleus muscle.
Muscle performance is improved after a brief period of exercise (warm-up). One factor that is known to strongly affect force production is the myoplasmic concentration of inorganic phosphate ([P(i)]). Improved performance after warm-up may therefore be due to a reduction of [P(i)]. Herein, we show that after a warm-up protocol (15 tetani at 2-s intervals), tetanic force is increased by approximately 6% (P < 0.05) and [P(i)] is almost halved (P < 0.05) in isolated mouse soleus muscle. A warm-up protocol with longer intervals (15 tetani at 5-s intervals) reduced tetanic force and did not alter [P(i)]. We conclude that a reduction of [P(i)] contributes to the force-potentiating effect of warm-up.


Dietary fruits and arthritis.
Arthritis is a global health concern affecting a significant proportion of the population and associated with reduced quality of life. Among the different forms of arthritis, osteoarthritis (OA) and rheumatoid arthritis (RA) are the most common and lacking a definite cure in the affected individuals. Fruits, such as berries and pomegranates are rich sources of a variety of dietary bioactive compounds, especially the polyphenolic flavonoids that have been associated with antioxidant, anti-inflammatory and analgesic effects. Emerging research demonstrates a protective role of fruits and their polyphenols in pre-clinical, clinical and epidemiological studies of OA and RA. In this context, commonly available fruits, such as blueberries, raspberries and strawberries, and pomegranates have shown promising results in reducing pain and inflammation in experimental models and in human clinical studies of arthritis. There is also some evidence on the role of specific fruit polyphenols, such as quercetin and citrus flavonoids in alleviating RA symptoms. These emerging data deserve further investigation in rigorous scientific studies to determine the mechanisms, dosing and selection of fruits and fruit extracts in arthritis management.


Huntingtin-associated protein 1 (HAP1) interacts with the p150Glued subunit of dynactin.
Huntington's disease (HD) is an inherited neurodegenerative disease caused by expansion of a polyglutamine repeat in the HD protein huntingtin. Huntingtin's localization within the cell includes an association with cytoskeletal elements and vesicles. We previously identified a protein (HAP1) which binds to huntingtin in a glutamine repeat length-dependent manner. We now report that HAP1 interacts with cytoskeletal proteins, namely the p150 Glued subunit of dynactin and the pericentriolar protein PCM-1. Structural predictions indicate that both HAP1 and the interacting proteins have a high probability of forming coiled coils. We examined the interaction of HAP1 with p150 Glued . Binding of HAP1 to p150 Glued (amino acids 879-1150) was confirmed in vitro by binding of p150 Glued to a HAP1-GST fusion protein immobilized on glutathione-Sepharose beads. Also, HAP1 co-immunoprecipitated with p150 Glued from brain extracts, indicating that the interaction occurs in vivo . Like HAP1, p150 Glued is highly expressed in neurons in brain and both proteins are enriched in a nerve terminal vesicle-rich fraction. Double label immunofluorescence experiments in NGF-treated PC12 cells using confocal microscopy revealed that HAP1 and p150 Glued partially co-localize. These results suggest that HAP1 might function as an adaptor protein using coiled coils to mediate interactions among cytoskeletal, vesicular and motor proteins. Thus, HAP1 and huntingtin may play a role in vesicle trafficking within the cell and disruption of this function could contribute to the neuronal dysfunction and death seen in HD.


Prevalence of dysthymic disorder in primary care.
Dysthymic disorder is characterised as a chronic state of depressed mood which is not otherwise attributable to physical, psychological or social events. While it can occur alone, there is increasing evidence that the majority of individuals who meet criteria for dysthymic disorder also experience more severe episodic mood disorders throughout their lifetime, and there is also an aggregation of mood disorders within their family members. Patients with dysthymic disorder are most often seen in primary care. Some researchers suggest that the majority of these individuals are never diagnosed or are not diagnosed until a more severe episodic mood disorder develops. The objective of this study was to determine the 12-month prevalence of Axis I psychiatric disorders, and in particular dysthymic disorder, in a primary care Health Service Organization in Ontario, Canada. Eligible and consenting adults registered with a primary care Health Service Organization were screened using the modified form of the University of Michigan Composite International Diagnostic Interview. Of the 6280 eligible subjects, 4327 (69%) consented to screening. Two hundred and twenty-two (5.1%) subjects screened positive for dysthymic disorder. In addition, 90% of those who screened positive for dysthymic disorder also screened positive for other Axis I disorders including major depressive disorder, panic, simple phobia, and generalized anxiety disorder. There is much potential for the primary care physician to play a pivotal role in the recognition and treatment of dysthymic disorder and associated Axis I disorders. A focus on the family as a unit for care may be especially important given the reported aggregation of dysthymic disorder within families.


Peptide-based nanoprobes for molecular imaging and disease diagnostics.
Pathological changes in a diseased site are often accompanied by abnormal activities of various biomolecules in and around the involved cells. Identifying the location and expression levels of these biomolecules could enable early-stage diagnosis of the related disease, the design of an appropriate treatment strategy, and the accurate assessment of the treatment outcomes. Over the past two decades, a great diversity of peptide-based nanoprobes (PBNs) have been developed, aiming to improve the in vitro and in vivo performances of water-soluble molecular probes through engineering of their primary chemical structures as well as the physicochemical properties of their resultant assemblies. In this review, we introduce strategies and approaches adopted for the identification of functional peptides in the context of molecular imaging and disease diagnostics, and then focus our discussion on the design and construction of PBNs capable of navigating through physiological barriers for targeted delivery and improved specificity and sensitivity in recognizing target biomolecules. We highlight the biological and structural roles that low-molecular-weight peptides play in PBN design and provide our perspectives on the future development of PBNs for clinical translation.


A microplate solid scintillation counter as a radioactivity detector for high performance liquid chromatography in drug metabolism: validation and applications.
Sensitive radioactivity detection following high performance liquid chromatography (HPLC) separation remains a challenge in many drug metabolism studies with radiolabeled compounds. In this work, solid scintillation counting (SSC) after fraction collection into 96-well plates was evaluated as an off-line radioactivity detection method, in comparison with conventional liquid scintillation counting (LSC). The impact of counting time and biological matrix on the quantification of radiolabeled metabolites and parent drug in samples from animal and human absorption, distribution, metabolism and excretion (ADME) studies was investigated. Three different approaches were used to test whether reliable quantification by off-line SSC detection, which requires an approximately constant counting yield during the entire chromatographic run, can be realized: (i) the measurement of radioactivity-spiked biological blank samples without HPLC separation as an extreme case of biological background, (ii) the measurement of radioactivity-spiked HPLC fractions of biological blank samples and (iii) the comparison of radiochromatograms obtained by off-line SSC and LSC of real samples from ADME studies with radiolabeled compounds. Situations in which variations in SSC yield during an HPLC run are likely to lead to significant errors in quantitation were identified and are discussed. However, examples from a number of animal or human ADME studies showed that in the majority of cases off-line SSC provides very similar quantitative data, compared with the reference method of off-line LSC radioactivity detection. Approaches for validation of the off-line SSC approach in critical cases are discussed. The main advantages of off-line SSC, compared with off-line LSC, are lower detection limits and a substantially higher throughput. Several applications of off-line SSC detection in ADME studies are shown.


Effects of obesogenic diet and estradiol on dorsal raphe gene expression in old female macaques.
The beneficial effects of bioidentical ovarian steroid hormone therapy (HT) during the perimenopause are gaining recognition. However, the positive effects of estrogen (E) plus or minus progesterone (P) administration to ovariectomized (Ovx) lab animals were recognized in multiple systems for years before clinical trials could adequately duplicate the results. Moreover, very large numbers of women are often needed to find statistically significant results in clinical trials of HT; and there are still opposing results being published, especially in neural and cardiovascular systems. One of the obvious differences between human and animal studies is diet. Laboratory animals are fed a diet that is low in fat and refined sugar, but high in micronutrients. In the US, a large portion of the population eats what is known as a "western style diet" or WSD that provides calories from 36% fat, 44% carbohydrates (includes 18.5% sugars) and 18% protein. Unfortunately, obesity and diabetes have reached epidemic proportions and the percentage of obese women in clinical trials may be overlooked. We questioned whether WSD and obesity could decrease the positive neural effects of estradiol (E) in the serotonin system of old macaques that were surgically menopausal. Old ovo-hysterectomized female monkeys were fed WSD for 2.5 years, and treated with placebo, Immediate E (ImE) or Delayed E (DE). Compared to old Ovx macaques on primate chow and treated with placebo or E, the WSD-fed monkeys exhibited greater individual variance and blunted responses to E-treatment in the expression of genes related to serotonin neurotransmission, CRH components in the midbrain, synapse assembly, DNA repair, protein folding, ubiquitylation, transport and neurodegeneration. For many of the genes examined, transcript abundance was lower in WSD-fed than chow-fed monkeys. In summary, an obesogenic diet for 2.5 years in old surgically menopausal macaques blunted or increased variability in E-induced gene expression in the dorsal raphe. These results suggest that with regard to function and viability in the dorsal raphe, HT may not be as beneficial for obese women as normal weight women.


Naturally occurring antibodies that recognize linear epitopes in the amino terminus of the hepatitis C virus E2 protein confer noninterfering, additive neutralization.
Chronic hepatitis C virus (HCV) infection can persist even in the presence of a broadly neutralizing antibody response. Various mechanisms that underpin viral persistence have been proposed, and one of the most recently proposed mechanisms is the presence of interfering antibodies that negate neutralizing responses. Specifically, it has been proposed that antibodies targeting broadly neutralizing epitopes located within a region of E2 encompassing residues 412 to 423 can be inhibited by nonneutralizing antibodies binding to a less conserved region encompassing residues 434 to 446. To investigate this phenomenon, we characterized the neutralizing and inhibitory effects of human-derived affinity-purified immunoglobulin fractions and murine monoclonal antibodies and show that antibodies to both regions neutralize HCV pseudoparticle (HCVpp) and cell culture-infectious virus (HCVcc) infection albeit with different breadths and potencies. Epitope mapping revealed the presence of overlapping but distinct epitopes in both regions, which may explain the observed differences in neutralizing phenotypes. Crucially, we failed to demonstrate any inhibition between these two groups of antibodies, suggesting that interference by nonneutralizing antibodies, at least for the region encompassing residues 434 to 446, does not provide a mechanism for HCV persistence in chronically infected individuals.


Obstructive sleep apnea and cancer: Epidemiologic links and theoretical biological constructs.
Sleep disorders have emerged as highly prevalent conditions in the last 50-75 y. Along with improved understanding of such disorders, the realization that perturbations in sleep architecture and continuity may initiate, exacerbate or modulate the phenotypic expression of multiple diseases including cancer has gained increased attention. Furthermore, the intermittent hypoxia that is attendant to sleep disordered breathing, has recently been implicated in increased incidence and more adverse prognosis of cancer. The unifying conceptual framework linking these associations proposes that increased sympathetic activity and/or alterations in immune function, particularly affecting innate immune cellular populations, underlie the deleterious effects of sleep disorders on tumor biology. In this review, the epidemiological evidence linking disrupted sleep and intermittent hypoxia to oncological outcomes, and the potential biological underpinnings of such associations as illustrated by experimental murine models will be critically appraised. The overarching conclusion appears supportive in the formulation of an hypothetical framework, in which fragmented sleep and intermittent hypoxia may promote changes in multiple signalosomes and transcription factors that can not only initiate malignant transformation, but will also alter the tumor microenvironment, disrupt immunosurveillance, and thus hasten tumor proliferation and increase local and metastatic invasion. Future bench-based experimental studies as well as carefully conducted and controlled clinical epidemiological studies appear justified for further exploration of these hypotheses.


Characterization of a novel interaction between transcription factor TFII-I and the inducible tyrosine kinase in T cells.
TCR signaling leads to the activation of kinases such as inducible tyrosine kinase (Itk), a key regulatory protein in T-lymphocyte activation and function. The homolog of Itk in B cells is Bruton's tyrosine kinase, previously shown to bind and phosphorylate the transcription factor TFII-I. TFII-I plays major roles in transcription and signaling. Our purpose herein was twofold: first, to identify some of the molecular determinants involved in TFII-I activation downstream of receptor crosslinking in T cells and second, to uncover the existence of Itk-TFII-I signaling in T lymphocytes. We report for the first time that TFII-I is tyrosine phosphorylated upon TCR, TCR/CD43, and TCR/CD28 co-receptor engagement in human and/or murine T cells. We show that Itk physically interacts with TFII-I and potentiates TFII-I-driven c-fos transcription. We demonstrate that TFII-I is phosphorylated upon co-expression of WT, but not kinase-dead, or kinase-dead/R29C mutant Itk, suggesting these residues are important for TFII-I phosphorylation, presumably via an Itk-dependent mechanism. Structural analysis of TFII-I-Itk interactions revealed that the first 90 residues of TFII-I are dispensable for Itk binding. Mutations within Itk's kinase, pleckstrin-homology, and proline-rich regions did not abolish TFII-I-Itk binding. Our results provide an initial step in understanding the biological role of Itk-TFII-I signaling in T-cell function.


Diseases Transmitted by Man's Best Friend: The Dog.
The relationship between dogs and humans is ancient and mutually beneficial. Dogs have served people well as companions, workmates, guides, and protectors. However, on occasion, dogs may injure humans through biting or may transmit pathogens resulting in a large number of problems ranging from a trivial rash to life-threatening bacteremia. Given that there are more than 80 million pet dogs in the United States, it is worth knowing the potential problems that can result from canine exposure. Annually, almost 5 million people in the United States suffer a dog bite. Dog bite wounds become infected up to 15% of the time. In those who have had a splenectomy, a dog bite may transmit the bacterium Capnocytophaga canimorsus, leading to life-threatening bacteremia. Other illnesses that humans can acquire from dog contact include ringworm, diarrheal disease (salmonellosis, campylobacteriosis, and intestinal parasites), leptospirosis, brucellosis, Q fever, visceral larva migrans, and echinococcosis. Evidence exists that the family dog may serve as a reservoir for uropathogenic Escherichia coli that can lead to urinary tract infections among human household contacts. In this article we discuss dog-related infectious diseases as well as measures to minimize dog-associated illness (e.g., do not disturb sleeping dogs; HIV-infected persons who wish to acquire a puppy should have the dog's stool checked for Cryptosporidium).


CD31/PECAM-1-driven chemokine-independent transmigration of human T lymphocytes.
We assessed the relative contribution of CD31/PECAM-1 (platelet-endothelial cell adhesion molecule-1) to T lymphocyte transmigration by the use of transfected murine fibroblasts stably expressing either the human CD31/PECAM-1 or the intercellular adhesion molecule-1 (CD54/ICAM-1). Unlike CD54/ICAM-1, CD31/PECAM-1 supported migration of activated T cells in the absence of chemokines: most of the migrating lymphocytes were CD31+ and displayed a phenotype corresponding to the naive subpopulation (LFA-1 dull and CD45RA+). Migration of activated T lymphocytes through CD54/ICAM-1+ transfected monolayers could be induced by creating a chemotactic gradient with the chemokine monocyte chemotactic protein-1, and the migrating cells mainly displayed a memory phenotype (LFA-1 bright CD45RO+) under these conditions. Furthermore, we found that in transfected cells CD54/ICAM-1 is uniformly distributed along the apical surface of the cells, while CD31/PECAM-1 is concentrated at the intercellular junctions, suggesting the existence of a haptotactic gradient (i.e. a gradient of substrate- or cell-bound molecules) responsible for T cell migration. This was also confirmed by the finding that monolayers of murine fibroblasts transfected with a CD31/PECAM-1 mutant lacking the cytoplasmic domain (CD31/PECAM-1-delta cyto), which has a reduced tendency to localize at cell-cell contact areas, supported efficient adhesion but were unable to induce migration of activated T cells unless a chemotactic gradient was created. We propose that in lymphocytes, homophilic CD31/PECAM-1 adhesion may be primarily involved in transmigration of naive T cells and that its role is complementary to that of CD54/ICAM-1.


Predicting potential responses to future climate in an alpine ungulate: interspecific interactions exceed climate effects.
The altitudinal shifts of many montane populations are lagging behind climate change. Understanding habitual, daily behavioural rhythms, and their climatic and environmental influences, could shed light on the constraints on long-term upslope range-shifts. In addition, behavioural rhythms can be affected by interspecific interactions, which can ameliorate or exacerbate climate-driven effects on ecology. Here, we investigate the relative influences of ambient temperature and an interaction with domestic sheep (Ovis aries) on the altitude use and activity budgets of a mountain ungulate, the Alpine chamois (Rupicapra rupicapra). Chamois moved upslope when it was hotter but this effect was modest compared to that of the presence of sheep, to which they reacted by moving 89-103 m upslope, into an entirely novel altitudinal range. Across the European Alps, a range-shift of this magnitude corresponds to a 46% decrease in the availability of suitable foraging habitat. This highlights the importance of understanding how factors such as competition and disturbance shape a given species' realised niche when predicting potential future responses to change. Furthermore, it exposes the potential for manipulations of species interactions to ameliorate the impacts of climate change, in this case by the careful management of livestock. Such manipulations could be particularly appropriate for species where competition or disturbance already strongly restricts their available niche. Our results also reveal the potential role of behavioural flexibility in responses to climate change. Chamois reduced their activity when it was warmer, which could explain their modest altitudinal migrations. Considering this behavioural flexibility, our model predicts a small 15-30 m upslope shift by 2100 in response to climate change, less than 4% of the altitudinal shift that would be predicted using a traditional species distribution model-type approach (SDM), which assumes that species' behaviour remains unchanged as climate changes. Behavioural modifications could strongly affect how species respond to a changing climate.


[Neurofeedback in the Treatment of Anorexia Nervosa: a Case Report].
Anorexia nervosa has been related to alterations in brain activity in terms of hyperactive EEG patterns. This case report illustrates the principles and results of a five-week neurofeedback treatment in a 29-year-old woman suffering from anorexia nervosa. A neurofeedback protocol to enhance alpha activity (8 - 12 Hz) was developed and conducted additionally to the standardized treatment for eating disorders in training sessions twice a week. Pre- and post-test measurements included resting state EEG measurements and a psychological test battery. The results show improvements from pre- to post-test in eating disorder psychopathology including psychological wellbeing, emotional competence, and eating behavior traits. In addition, a decrease in theta power (4 - 7 Hz), a well-known trait marker of anorexia nervosa, was measured. However, our data should be interpreted with caution because this is a single case study. Nevertheless, this report documents the practicability and method of neurofeedback as treatment adjunct in eating disorders from the clinical perspective. Although the use of neurofeedback in the treatment of anorexia nervosa is recommended in literature, empirical studies are still lacking. Randomized controlled trials to evaluate short- and long-term effects of neurofeedback are needed.


EEG alpha oscillations during the performance of verbal creativity tasks: differential effects of sex and verbal intelligence.
Task-related power changes in the EEG alpha band were analyzed in 31 participants (17 males and 14 females) during performance of two verbal creativity tasks. Participants were confronted with verbal problems that are in need of explanation (insight problems) and utopian situations that will actually never happen. In both tasks they were instructed to generate as many but also as unusual, unique or original ideas as possible. To assess brain responses that come along with highly original ideas, individual responses were divided into more and less original ideas (within each participant). Creative problem solving was generally accompanied by lower levels of cortical arousal (i.e., increases in alpha power from a pre-stimulus reference to an activation interval). Additionally, more original (vs. less original) responses were associated with a stronger task-related alpha synchronization in posterior (particularly centroparietal) cortices. Task-related alpha power changes during creative problem solving were also moderated by verbal IQ and sex.


Displacement of carbonates in Ca2UO2(CO3)3 by amidoxime-based ligands from free-energy simulations.
Amidoxime-based ligands are effective in uranium extraction by displacing carbonates in Ca2UO2(CO3)3, the dominant uranyl species in seawater. However, a detailed understanding of the displacement process has been lacking. Here we use classical molecular dynamics combined with umbrella sampling to map the complete displacement process and the free-energy profiles by the simple acetamidoximate (AO-) and the more complex glutardiamidoximate (B2- and HB-) ligands. Interestingly, we find that the two Ca2+ ions in Ca2UO2(CO3)3 can greatly facilitate the displacement of the first two carbonate groups. Displacing the third carbonate is however significantly more uphill than the first two. With the help of an additional Ca2+ ion, the third carbonate displacement can be made less uphill. Comparing AO- and B2-/HB- ligands, we find that the displacement by the latter is thermodynamically more favorable due to the chelate effect. Our free-energy simulations based on classical molecular dynamics simulations reveal key atomistic details and quantify the thermodynamic driving force during the carbonate displacement of Ca2UO2(CO3)3 by amidoxime-based ligands. These findings will be useful in understanding seawater uranium extraction by amidoxime-grafted polymeric sorbents.


High-dose chemotherapy with hematopoietic stem cell transplantation is effective for nasal and nasal-type CD56+ natural killer cell lymphomas.
CD56+ natural killer (NK) cell lymphomas occur frequently in the nasal and nasopharyngeal regions and carry a poor prognosis. We have studied seven cases with NK-cell lymphomas. These lymphomas showed the following immunophenotype: CD56+, CD2+, sCD3- and Epstein-Barr virus-encoded small RNAs (EBERs)+. Six patients had localized (stage I or II) disease involving the nasopharyngeal region, while one had stage III disease. One patient with stage I disease achieved a complete remission (CR) after treatment with involved-field irradiation, but subsequently relapsed and died. The remaining six patients received combination chemotherapy as primary treatment: five patients with localized stage I or II disease and one patient with advanced stage III disease. Responses to initial chemotherapy were generally poor. These six patients received a variety of salvage chemotherapy regimens, but never achieved a CR. Subsequently, four of six patients showed a highly aggressive clinical course and died of disseminated disease within 1 year from the diagnosis. Three of six patients received high-dose chemotherapy supported by syngeneic, autologous or allogeneic peripheral blood stem cell transplantation. Two of the three transplant patients achieved a CR and are now surviving in continuous CR. Our clinical experience suggests that myeloablative high-dose chemotherapy and bone marrow rescue by hematopoietic stem cell transplantation may be an effective salvage treatment modality for refractory NK-cell lymphomas and could be considered as a part of the initial therapy for these patients.


Family structure and dependency: early transitions to female household headship.
This article looks at the intergenerational effects of family structure on family formation and economic dependency. Three explanations are examined: the economic-deprivation, socialization, and family-stress hypotheses. Daughters living in single-parent families at some point during adolescence are more likely to become household heads and to go on welfare than offspring of two-parent families. Differences in the incomes of one- and two-parent families can account for up to 25 percent of the difference in offspring behaviors. None of the hypotheses, however, provides a complete explanation of family structure effects. The analysis is based on data from the Panel Study of Income Dynamics and uses event-history analysis to estimate transitions into female headship and economic dependence.


TIGIT Marks Exhausted T Cells, Correlates with Disease Progression, and Serves as a Target for Immune Restoration in HIV and SIV Infection.
HIV infection induces phenotypic and functional changes to CD8+ T cells defined by the coordinated upregulation of a series of negative checkpoint receptors that eventually result in T cell exhaustion and failure to control viral replication. We report that effector CD8+ T cells during HIV infection in blood and SIV infection in lymphoid tissue exhibit higher levels of the negative checkpoint receptor TIGIT. Increased frequencies of TIGIT+ and TIGIT+ PD-1+ CD8+ T cells correlated with parameters of HIV and SIV disease progression. TIGIT remained elevated despite viral suppression in those with either pharmacological antiretroviral control or immunologically in elite controllers. HIV and SIV-specific CD8+ T cells were dysfunctional and expressed high levels of TIGIT and PD-1. Ex-vivo single or combinational antibody blockade of TIGIT and/or PD-L1 restored viral-specific CD8+ T cell effector responses. The frequency of TIGIT+ CD4+ T cells correlated with the CD4+ T cell total HIV DNA. These findings identify TIGIT as a novel marker of dysfunctional HIV-specific T cells and suggest TIGIT along with other checkpoint receptors may be novel curative HIV targets to reverse T cell exhaustion.


Transfusion-Associated Circulatory Overload: A Clinical Perspective.
For 30 years, transfusion-associated circulatory overload (TACO) has been recognized as a serious transfusion complication. Currently, TACO is the leading cause of transfusion-related morbidity and mortality worldwide which occurs in 1% to 12% of at-risk populations. Despite an incomplete understanding of the underlying pathophysiology, TACO is defined as a collection of signs and symptoms of acute pulmonary edema due to circulatory overload occurring within 6 to 12 hours of transfusion. In the past decade, large observational cohort studies resulted in better insight into the associated transfusion risk factors leading to the development of TACO. In this clinical review, we critically analyze the pathogenesis of TACO, associated risk factors, clinical presentation, diagnostic modalities, and treatment options to guide clinicians with early detection of this syndrome and intervention to improve clinical outcomes. Future research should focus on better understanding of the pathogenesis to help advance the field of volume kinetics and endothelial barrier function.


SNP prioritization using a Bayesian probability of association.
Prioritization is the process whereby a set of possible candidate genes or SNPs is ranked so that the most promising can be taken forward into further studies. In a genome-wide association study, prioritization is usually based on the P-values alone, but researchers sometimes take account of external annotation information about the SNPs such as whether the SNP lies close to a good candidate gene. Using external information in this way is inherently subjective and is often not formalized, making the analysis difficult to reproduce. Building on previous work that has identified 14 important types of external information, we present an approximate Bayesian analysis that produces an estimate of the probability of association. The calculation combines four sources of information: the genome-wide data, SNP information derived from bioinformatics databases, empirical SNP weights, and the researchers' subjective prior opinions. The calculation is fast enough that it can be applied to millions of SNPS and although it does rely on subjective judgments, those judgments are made explicit so that the final SNP selection can be reproduced. We show that the resulting probability of association is intuitively more appealing than the P-value because it is easier to interpret and it makes allowance for the power of the study. We illustrate the use of the probability of association for SNP prioritization by applying it to a meta-analysis of kidney function genome-wide association studies and demonstrate that SNP selection performs better using the probability of association compared with P-values alone.


International aspirations for speech-language pathologists' practice with multilingual children with speech sound disorders: development of a position paper.
A major challenge for the speech-language pathology profession in many cultures is to address the mismatch between the "linguistic homogeneity of the speech-language pathology profession and the linguistic diversity of its clientele" (Caesar & Kohler, 2007, p. 198). This paper outlines the development of the Multilingual Children with Speech Sound Disorders: Position Paper created to guide speech-language pathologists' (SLPs') facilitation of multilingual children's speech. An international expert panel was assembled comprising 57 researchers (SLPs, linguists, phoneticians, and speech scientists) with knowledge about multilingual children's speech, or children with speech sound disorders. Combined, they had worked in 33 countries and used 26 languages in professional practice. Fourteen panel members met for a one-day workshop to identify key points for inclusion in the position paper. Subsequently, 42 additional panel members participated online to contribute to drafts of the position paper. A thematic analysis was undertaken of the major areas of discussion using two data sources: (a) face-to-face workshop transcript (133 pages) and (b) online discussion artifacts (104 pages). Finally, a moderator with international expertise in working with children with speech sound disorders facilitated the incorporation of the panel's recommendations. The following themes were identified: definitions, scope, framework, evidence, challenges, practices, and consideration of a multilingual audience. The resulting position paper contains guidelines for providing services to multilingual children with speech sound disorders (http://www.csu.edu.au/research/multilingual-speech/position-paper). The paper is structured using the International Classification of Functioning, Disability and Health: Children and Youth Version (World Health Organization, 2007) and incorporates recommendations for (a) children and families, (b) SLPs' assessment and intervention, (c) SLPs' professional practice, and (d) SLPs' collaboration with other professionals. Readers will 1. recognize that multilingual children with speech sound disorders have both similar and different needs to monolingual children when working with speech-language pathologists. 2. Describe the challenges for speech-language pathologists who work with multilingual children. 3. Recall the importance of cultural competence for speech-language pathologists. 4. Identify methods for international collaboration and consultation. 5. Recognize the importance of engaging with families and people within their local communities for supporting multilingual children in context.


[Pulmonary lesions and total parenteral nutrition in children admitted to a pediatric intensive care unit].
To describe lung injuries in autopsied pediatric patients (neonates through 15 years old) subjected or not to total parenteral nutrition, in an intensive care unit. Sections from six paraffin-embedded lung fragments, from 114 children were studied by routine staining. Demographic, clinical and therapeutic data were retrieved from the records. Statistical analysis was performed using Statistical Package for the Social Sciences. The 114 patients were divided in two groups: 50 who were treated with total parenteral nutrition with lipid emulsion and 64 who did not receive total parenteral nutrition. The two groups did not differ in gender (p = 0.654), age (p = 0.682) or body weight (p = 0.175), but duration of hospital stay (p = 0.000), prematurity (p = 0.008) and treatment with blood products (p = 0.009) were all higher in the group treated with total parenteral nutrition. All patients received mechanical ventilation during hospitalization. Chi-square comparisons showed that diffuse alveolar injury (p = 0.022), pulmonary fibrosis (p = 0.019), pneumocyte hyperplasia (p = 0.004), microthromboembolism (p = 0.047) and thrombophlebitis (0.033) all exhibited a significant relationship with total parenteral nutrition. However, a multivariate analysis by logistic regression, taking into account prematurity and duration of hospital stay, demonstrated that total parenteral nutrition was an independent factor only with respect of pulmonary fibrosis. Although lung injuries were significantly more frequent in children who had received total parenteral nutrition, it was impossible to conclude that the lipid infusion had a direct relationship with these injuries, because prematurity and duration of hospital stay were significant co-factors.


Diabetes (db/db) mutation-induced endometrial epithelial lipoapoptosis: ultrastructural and cytochemical analysis of reproductive tract atrophy.
The diabetes (db/db) mutation in C57BL/KsJ mice promotes a progressive cytolipidemia within the endometrial epithelial (EE) layer of the female reproductive tract which results in premature cellular and organ atrophy. The current studies focus on the ultrastructural and cytochemical changes which promote nuclear apoptosis and cytostructural disruption following the expression of endometrial hypercytolipidemia which promotes diabetes-associated organoinvolution and manifest infertility. Control (normal:+/+) and diabetes (db/db) genotype groups were prepared for high resolution light microscopic analysis of cytolipidemia and nuclear apoptosis (TUNEL-labeled 3'-DNA fragmentation) indices and compared to the transmission electron (TEM) microscopic analysis of endometrial tissue samples collected from 8-16 week-old groups. Compared to controls, db/db mutation expression induced a dramatic increase in EE cytolipid vacuole volume and density within the epithelial endometrial layer. TEM analysis revealed that cytolipid vacuole accumulations initially aggregated at the baso-polar regions of UEE cells in response to the systemic hyperglycemic/hypertriglyceridemic conditions which characterized the (db/db) groups. Progressive cytoplasmic movement of the lipid pools into perinuclear compartments of affected EE cells induced nuclear isolation from organelles that were displaced towards peripheral cytoplasmic compartments. Cytochemical analysis of lipid vacuole accumulations indicated attraction towards, and incorporation within, the nuclear envelope of hyperlipidemic cells. Co-localization of nuclear apoptotic 3'-DNA fragments within identified hyperlipidemic EE cells was coincident with the cytochemical and ultrastructural identification of lipid penetration through the nuclear envelope in db/db mutants. These results are the first cytochemical indication that the metabolic disturbances in db/db mutants which promote hypercytolipidemia are coincident with lipoapoptosis-induced nuclear dissolution, as denoted by DNA fragmentation analysis. The lipidemia-induced alterations in intracellular organelle and nuclear architectures suggests that the metabolic disturbances in glucose and lipid metabolic cascades in diabetes (db/db) mutants disrupts cytointegrity, culminating in nuclear disregulation (as indicated by lipoapoptosis) and eventual premature reproductive tract organoinvolution and resultant, manifest, reproductive sterility.


Strengthening the implementation of Health in All Policies: a methodology for realist explanatory case studies.
To address macro-social and economic determinants of health and equity, there has been growing use of intersectoral action by governments around the world. Health in All Policies (HiAP) initiatives are a special case where governments use cross-sectoral structures and relationships to systematically address health in policymaking by targeting broad health determinants rather than health services alone. Although many examples of HiAP have emerged in recent decades, the reasons for their successful implementation--and for implementation failures--have not been systematically studied. Consequently, rigorous evidence based on systematic research of the social mechanisms that have regularly enabled or hindered implementation in different jurisdictions is sparse. We describe a novel methodology for explanatory case studies that use a scientific realist perspective to study the implementation of HiAP. Our methodology begins with the formulation of a conceptual framework to describe contexts, social mechanisms and outcomes of relevance to the sustainable implementation of HiAP. We then describe the process of systematically explaining phenomena of interest using evidence from literature and key informant interviews, and looking for patterns and themes. Finally, we present a comparative example of how Health Impact Assessment tools have been utilized in Sweden and Quebec to illustrate how this methodology uses evidence to first describe successful practices for implementation of HiAP and then refine the initial framework. The methodology that we describe helps researchers to identify and triangulate rich evidence describing social mechanisms and salient contextual factors that characterize successful practices in implementing HiAP in specific jurisdictions. This methodology can be applied to study the implementation of HiAP and other forms of intersectoral action to reduce health inequities involving multiple geographic levels of government in diverse settings.


New approaches for the prevention of rejection and graft-vs.-host disease in clinical bone marrow transplantation.
The two major barriers to successful allogeneic bone marrow transplantation (BMT) in animals and man are graft-vs.-host disease (GVHD) and the risk of graft rejection. GVHD is the result of alloreactivity of mature donor T-lymphocytes present in the graft-vs.-host tissues and can be completely prevented by pregraft depletion of T-lymphocytes. Graft rejection results from residual host immunocompetent lymphocytes that survive heavy chemoradiotherapy prior to allogeneic BMT. Host resistance to allograft cannot be eradicated even by conventional conditioning with high-dose cyclophosphamide (120 mg/kg) and lethal whole body irradiation (1,200 rad). In the present report we have utilized two new techniques to overcome GVHD and graft rejection following allogeneic BMT. GVHD can be prevented by a new monoclonal rat antihuman lymphocyte antibody, CAMPATH-1, which binds human complement, enabling donor serum to serve as the source of complement. Prevention of rejection of T-lymphocyte-depleted marrow allografts can be achieved by the application of total lymphoid irradiation (TLI) in addition to conventional chemoradiotherapy, prior to allogeneic BMT. TLI causes potent immunosuppression with minimal side effects. A combination of TLI for overcoming host resistance to allograft, and CAMPATH-1 for overcoming GVHD, leads to a relatively smooth posttransplant outcome with no evidence of GVHD and with no need for posttransplant immunosuppression.


Pressure ulcers and pressure relief surfaces.
Pressure ulcer incidence has been reduced but not eliminated. A few patients still develop pressure ulcers despite using air-fluidized beds and other specialty beds. In the future, very high-risk patients may possibly be sent to space clinics to recuperate in zero gravity for extended periods. Another possibility is the creation of suspension devices to keep a patient off the areas of the body where pressure sores might occur. Being informed of what is available and and aware how to use it optimally is currently the best solution.