Treatment for BK virus: incidence, risk factors and outcomes for kidney transplant recipients in the United States.
There has been a notable rise of BK virus among kidney transplant recipients. Single-center reports have identified risk factors for development of BK virus. However, there has not been an assessment of risk factors and incidence of this complication at a national level. This study utilized newly collected follow-up information from the national SRTR database to investigate incidence, risk factors and outcomes for solitary kidney transplant recipients associated with treatment for BK virus (TBKV) from 2004 to 2006. Logistic and Cox models were utilized to assess risk factors and evaluate graft survival associated with TBKV. Incidence of TBKV was 1.6% at 6 months and 2.6% at 1 year following transplantation. Patients with and without TBKV at 6 months had 79% and 90% 3-year overall graft survival respectively. Risk factors included advanced donor age, pediatric, African American and male recipients, human leukocyte antigen-mismatching and tacrolimus and thymoglobulin induction as baseline immunosuppression. Acute rejection episodes were more frequent prior to and following TBKV. TBKV is a common and rising incidence, varies based on transplant characteristics and should be included as a safety endpoint in studies investigating immunosuppressive protocols. Careful monitoring and further understanding of disease etiology and treatment strategies are needed.


Angiotensin-converting enzyme inhibitors: considerations regarding proteinuria.
The effects of captopril on renal function are evaluated in hypertensive patients with impaired renal function previously entered in a worldwide surveillance study. For the overwhelming majority of patients, control of BP was accompanied by stable or improved renal function. To date, the major adverse renal reactions have been a somewhat increased risk of worsening proteinuria among these patients and the well-documented acute decrease in renal function that may occur in patients with bilateral renal artery stenosis.


Favorable impact of virological response to highly active antiretroviral therapy on survival in patients with AIDS-related lymphoma.
We have retrospectively studied the influence of highly active antiretroviral therapy (HAART) on the outcome of AIDS-related lymphomas (ARL) as well as the possible influence of the virological response to HAART on complete response (CR) rate and survival in our series of ARL treated with CHOP. Two groups of patients were studied: (1) 44 patients who did not take HAART when the lymphoma was diagnosed, and (2) 26 patients treated with HAART concomitantly and after chemotherapy. There were 4 (9%) women in group 1 versus 11 (42%) in group 2 (P = 0.01), and serum lactate dehydrogenase (LDH) level was lower in group 2. The response rate to CHOP was higher in group 2 patients (15 out of 23, 65%) than in those of group 1 (16 out of 44, 36%) (P = 0.025). The factors associated with improvement of CR in the multivariate analysis were the administration of HAART (P = 0.004) and International Prognostic Index (IPI) score < or = 2 (P = 0.006). Among group 2 patients, those with a virological response to HAART and with IPI score < or = 2 had better response rate to chemotherapy (odds ratios 9.3 and 11.8, respectively). The median (95% CI) overall survival (OS) for group 1 patients was 7 (3-11) months, whereas it has not been reached for group 2 (P = 0.002). The only parameters influencing OS in the multivariate analysis were HAART (0.003), as a protective factor and IPI score > 2 (P = 0.015) with negative influence. Among patients treated with HAART, those with virological response had higher OS probability (P = 0.004), whereas those with IPI score > 2 had an unfavorable prognosis (P = 0.014). The only variable with statistical significance for disease free survival (DFS) in the univariate and multivariate analyses was HAART (P = 0.0168 and P = 0.028, respectively). We conclude that HAART is an independent prognostic factor for CR attainment, OS and DFS in patients with ARL treated with CHOP. Those patients with virological response to HAART had a better survival.


The TRAIL of oncogenes to apoptosis.
Despite the significant advances in clinical research, surgical resection, radiotherapy and chemotherapy are still used as the primary method for cancer treatment. As compared to conventional therapies that often induce systemic toxicity and eventually contribute to tumor resistance, the TNF-related apoptosis-inducing ligand (TRAIL) is a promising anticancer agent that selectively triggers apoptosis in various cancer cells by interacting with its proapoptotic receptors DR4 and KILLER/DR5, while sparing the normal surrounding tissue. The intensive studies of TRAIL signaling pathways over the past decade have provided clues for understanding the molecular mechanisms of TRAIL-induced apoptosis in carcinogenesis and identified an array of therapeutic responses elicited by TRAIL and its receptor agonists. Analysis of its activity at the molecular level has shown that TRAIL improves survival either as monotherapies or combinatorial therapies with other mediators of apoptosis or anticancer chemotherapy. Combinatorial treatments amplify the activities of anticancer agents and widen the therapeutic window by overcoming tumor resistance to apoptosis and driving cancer cells to self-destruction. Although TRAIL sensitivity varies widely depending on the cell type, nontransformed cells are largely resistant to death mediated by TRAIL Death Receptors (DRs). Genetic alterations in cancer can contribute in tumor progression and often play an important role in evasion of apoptosis by tumor cells. Remarkably, RAS, MYC and HER2 oncogenes have been shown to sensitise tumor cells to TRAIL induced cell death. Here, we summarise the cross-talk of oncogenic and apoptotic pathways and how they can be exploited toward efficient combinatorial therapeutic protocols.


Acanthamoeba castellanii trophozoites escape killing by neutrophil extracellular traps using their 3'-nucleotidase/nuclease activity.
Acanthamoeba castellanii is a free-living amoeba that acts as an opportunistic pathogen for humans and is the pathogenic agent of Acanthamoeba keratitis (AK). A. castellanii may present as proliferative and infective trophozoites or as resistant cysts during their life cycle. The immune response against AK is still poorly explored; however, it is well established that macrophages and neutrophils play essential roles in controlling corneal infection during the disease outcome. The release of NETs is one of the innate immune strategies to prevent parasite infection, especially when neutrophils interact with microorganisms that are too large to be phagocytosed, which is the case for amoeba species. The present work demonstrated that A. castellanii trophozoites can trigger NET formation upon in vitro interaction with neutrophils. Using DNase as a control, we observed increased parasite survival after coinciding with neutrophils, which may be correlated with NET degradation. Indeed, A. castellanii trophozoites degrade the NET DNA scaffold. Molecular analysis confirmed the occurrence of a 3'-nucleotidase/nuclease (3'-NT/NU) in the A. castellanii genome. We also demonstrated that trophozoites exhibit significantly higher 3'-NT/NU activity than cysts, which cannot trigger NET release. Considering that previous studies indicated the pathological role of 3'-NT-/NU in parasite infection, we suggest that this enzyme may act as the mechanism of escape of A. castellanii trophozoites from NETs.


The relationship of cimetidine to 5-hydroxytryptamine tissue levels in stress ulceration in the rat.
We studied the effect of cimetidine on 5-hydroxytryptamine (5-HT) levels in various tissues in restrained rats. Sixty male Sprague-Dawley rats, divided into four groups, received four intraperitoneal injections at 6-hour intervals in doses determined by body weight. Group I (control) was injected with saline (1 ml/300 gm) and was not restrained. Group II was injected with saline (1 ml/300 gm) and restrained for 18 hours beginning 1 hour after the first injection. Group III was injected with cimetidine (100 mg/kg) and was not restrained, whereas Group IV was injected with cimetidine (100 mg/kg) and restrained according to the schedule for Group II. The rats were killed and their stomachs were collected and graded for ulceration. 5-Hydroxytryptamine levels in the antrum and fundus of the stomach, the colon, the liver, and rectus abdominis muscle were determined by spectrophotofluorometry. Cimetidine prevented an increase in 5-HT in the antrum and protected against ulceration. As 5-HT levels in other tissues were not altered by restraint-induced stress, the phenomenon may be specific to the antrum. The protection against ulceration and elevated 5-HT levels conferred by cimetidine was comparable to that of vagotomy in similar models.


Progression of spontaneous seizures after status epilepticus is associated with mossy fibre sprouting and extensive bilateral loss of hilar parvalbumin and somatostatin-immunoreactive neurons.
The development of spontaneous limbic seizures was investigated in a rat model in which electrical tetanic stimulation of the angular bundle was applied for up to 90 min. This stimulation produced behavioural and electrographic seizures that led to a status epilepticus (SE) in most rats (71%). Long-term EEG monitoring showed that the majority of the rats (67%) that underwent SE, displayed a progressive increase of seizure activity once the first seizure was recorded after a latent period of about 1 week. The other SE rats (33%) did not show this progression of seizure activity. We investigated whether these different patterns of evolution of spontaneous seizures could be related to differences in cellular or structural changes in the hippocampus. This was the case regarding the following changes. (i) Cell loss in the hilar region: in progressive SE rats this was extensive and bilateral whereas in nonprogressive SE rats it was mainly unilateral. (ii) Parvalbumin and somatostatin-immunoreactive neurons: in the hilar region these were almost completely eliminated in progressive SE rats but were still largely present unilaterally in nonprogressive SE rats. (iii) Mossy fibre sprouting: in progressive SE rats, extensive mossy fibre sprouting was prominent in the inner molecular layer. In nonprogressive SE rats, mossy fibre sprouting was also present but less prominent than in progressive SE rats. Although mossy fibre sprouting has been proposed to be a prerequisite for chronic seizure activity in experimental temporal lobe epilepsy, the extent of hilar cell death also appears to be an important factor that differentiates between whether or not seizure progression will occur.


Cerebellar volume as imaging outcome in progressive multiple sclerosis.
To assess whether cerebellar volumes changes could represent a sensitive outcome measure in primary-progressive MS. Changes in cerebellar volumes over one-year follow-up, estimated in 26 primary-progressive MS patients and 20 controls with Freesurfer longitudinal pipeline, were assessed using Wilcoxon test and tested for their correlation with disability worsening by a logistic regression. Clinical worsening was defined as EDSS score increase or change of >20% for 25-foot walk test or 9-hole peg test scores at follow-up. Sample sizes for given treatment effects and power were calculated. The findings were validated in an independent cohort of 20 primary-progressive MS patients. Significant changes were detected in brain T1 lesion volume (p<0.01), cerebellar T2 and T1 lesion volume (p<0.01 and p<0.05), cerebellar volume, cerebellar cortex volume, and cerebellar WM volume (p<0.001). Only cerebellar volume and cerebellar cortex volume percentage change were significantly reduced in clinically progressed patients when compared to patients who did not progress (p<0.01; respectively AUC of 0.91 and 0.96). Cerebellar volume percentage changes were consistent in the exploration and validation cohorts (cerebellar volume -1.90±1.11% vs -1.47±2.30%; cerebellar cortex volume -1.68±1.41% vs -1.56±2.23%). Based on our results the numbers of patients required to detect a 30% effect are 81 per arm for cerebellar volume and 162 per arm for cerebellar cortex volume (90% power, type 1 error alpha = 0.05). Our results suggest a role for cerebellar cortex volume and cerebellar volume as potential short-term imaging metrics to monitor treatment effect in primary-progressive MS clinical trials.


Anatomical networks reveal the musculoskeletal modularity of the human head.
Mosaic evolution is a key mechanism that promotes robustness and evolvability in living beings. For the human head, to have a modular organization would imply that each phenotypic module could grow and function semi-independently. Delimiting the boundaries of head modules, and even assessing their existence, is essential to understand human evolution. Here we provide the first study of the human head using anatomical network analysis (AnNA), offering the most complete overview of the modularity of the head to date. Our analysis integrates the many biological dependences that tie hard and soft tissues together, arising as a consequence of development, growth, stresses and loads, and motion. We created an anatomical network model of the human head, where nodes represent anatomical units and links represent their physical articulations. The analysis of the human head network uncovers the presence of 10 musculoskeletal modules, deep-rooted in these biological dependences, of developmental and evolutionary significance. In sum, this study uncovers new anatomical and functional modules of the human head using a novel quantitative method that enables a more comprehensive understanding of the evolutionary anatomy of our lineage, including the evolution of facial expression and facial asymmetry.


Concurrent intrathecal methotrexate and liposomal cytarabine for leptomeningeal metastasis from solid tumors: a retrospective cohort study.
Leptomeningeal metastasis (LM) from solid tumors is typically a late manifestation of systemic cancer with limited survival. Randomized trials comparing single agent intrathecal methotrexate to liposomal cytarabine have shown similar efficacy and tolerability. We hypothesized that combination intrathecal chemotherapy would be a safe and tolerable option in solid tumor LM. We conducted a retrospective cohort study of combination IT chemotherapy in solid tumor LM at a single institution between April 2010 and July 2012. In addition to therapies directed at active systemic disease, each subject received IT liposomal cytarabine plus IT methotrexate with dexamethasone premedication. Patient characteristics, survival outcomes and toxicities were determined by systematic chart review. Thirty subjects were treated during the study period. The most common cancer types were breast 15 (50 %), glioblastoma 6 (20 %), and lung 5 (17 %). Cytologic clearance was achieved in 6 (33 %). Median non-glioblastoma overall survival was 30.2 weeks (n = 18; range 3.9-73.4), and did not differ significantly by tumor type. Median time to neurologic progression was 7 weeks (n = 8; range 0.9-57), with 10 subjects (56 %) experiencing death from systemic disease without progression of LM. Age less than 60 was associated with longer overall survival (p = 0.01). Six (21 %) experienced grade III toxicities during treatment, most commonly meningitis 2 (7 %). Combination IT chemotherapy was feasible in this small retrospective cohort. Prospective evaluation is necessary to determine tolerability, the impact on quality of life and neurocognitive outcomes or any survival benefit when compared to single agent IT chemotherapy.


Deep RNA sequencing reveals dynamic regulation of myocardial noncoding RNAs in failing human heart and remodeling with mechanical circulatory support.
Microarrays have been used extensively to profile transcriptome remodeling in failing human heart, although the genomic coverage provided is limited and fails to provide a detailed picture of the myocardial transcriptome landscape. Here, we describe sequencing-based transcriptome profiling, providing comprehensive analysis of myocardial mRNA, microRNA (miRNA), and long noncoding RNA (lncRNA) expression in failing human heart before and after mechanical support with a left ventricular (LV) assist device (LVAD). Deep sequencing of RNA isolated from paired nonischemic (NICM; n=8) and ischemic (ICM; n=8) human failing LV samples collected before and after LVAD and from nonfailing human LV (n=8) was conducted. These analyses revealed high abundance of mRNA (37%) and lncRNA (71%) of mitochondrial origin. miRNASeq revealed 160 and 147 differentially expressed miRNAs in ICM and NICM, respectively, compared with nonfailing LV. Among these, only 2 (ICM) and 5 (NICM) miRNAs are normalized with LVAD. RNASeq detected 18 480, including 113 novel, lncRNAs in human LV. Among the 679 (ICM) and 570 (NICM) lncRNAs differentially expressed with heart failure, ≈10% are improved or normalized with LVAD. In addition, the expression signature of lncRNAs, but not miRNAs or mRNAs, distinguishes ICM from NICM. Further analysis suggests that cis-gene regulation represents a major mechanism of action of human cardiac lncRNAs. The myocardial transcriptome is dynamically regulated in advanced heart failure and after LVAD support. The expression profiles of lncRNAs, but not mRNAs or miRNAs, can discriminate failing hearts of different pathologies and are markedly altered in response to LVAD support. These results suggest an important role for lncRNAs in the pathogenesis of heart failure and in reverse remodeling observed with mechanical support.


Clinical review: Effect of vitamin D3 supplementation on improving glucose homeostasis and preventing diabetes: a systematic review and meta-analysis.
Observational studies report consistent associations between low vitamin D concentration and increased glycemia and risk of type 2 diabetes, but results of randomized controlled trials (RCTs) are mixed. The objective of the study was to systematically review RCTs that report on the effects of vitamin D supplementation on glucose homeostasis or diabetes prevention. Sources of data for the study were MEDLINE, EMBASE, SCOPUS, Cochrane Database of Systematic Reviews, Database of Abstracts of Reviews of Effects, Health Technology Assessment, and Science Citation Index from inception to June 2013. Study selection was trials that compared vitamin D3 supplementation with placebo or a non-vitamin D supplement in adults with normal glucose tolerance, prediabetes, or type 2 diabetes. Two reviewers collected data and assessed trial quality using the Cochrane Risk of Bias tool. Random-effects models were used to estimate mean differences (MDs) and odds ratios. The main outcomes of interest were homeostasis model assessment of insulin resistance, homeostasis model assessment of β-cell function, hemoglobin A1c levels, fasting blood glucose, incident diabetes, and adverse events. Thirty-five trials (43 407 patients) with variable risk of bias were included. Vitamin D had no significant effects on insulin resistance [homeostasis model assessment of insulin resistance: MD -0.04; 95% confidence interval (CI) -0.30 to 0.22, I-squared statistic (I(2)) = 45%], insulin secretion (homeostasis model of β-cell function: MD 1.64; 95% CI -25.94 to 29.22, I(2) = 40%), or hemoglobin A1c (MD -0.05%; 95% CI -0.12 to 0.03, I(2) = 55%) compared with controls. Four RCTs reported on the progression to new diabetes and found no effect of vitamin D (odds ratio 1.02; 95% CI 0.94 to 1.10, I(2) = 0%). Adverse events were rare, and there was no evidence of publication bias. Evidence from available trials shows no effect of vitamin D3 supplementation on glucose homeostasis or diabetes prevention. Definitive conclusions may be limited in the context of the moderate degree of heterogeneity, variable risk of bias, and short-term follow-up duration of the available evidence to date.


Individuality and adaptation across levels of selection: how shall we name and generalize the unit of Darwinism?
Two major clarifications have greatly abetted the understanding and fruitful expansion of the theory of natural selection in recent years: the acknowledgment that interactors, not replicators, constitute the causal unit of selection; and the recognition that interactors are Darwinian individuals, and that such individuals exist with potency at several levels of organization (genes, organisms, demes, and species in particular), thus engendering a rich hierarchical theory of selection in contrast with Darwin's own emphasis on the organismic level. But a piece of the argument has been missing, and individuals at levels distinct from organisms have been denied potency (although granted existence within the undeniable logic of the theory), because they do not achieve individuality with the same devices used by organisms and therefore seem weak by comparison. We show here that different features define Darwinian individuality across scales of size and time. In particular, species-individuals may develop few emergent features as direct adaptations. The interactor approach works with emergent fitnesses, not with emergent features; and species, as a consequence of their different mechanism for achieving individuality (reproductive exclusivity among subparts, that is, among organisms), express many effects from other levels. Organisms, by contrast, suppress upwardly cascading effects, because the organismic style of individuality (by functional integration of subparts) does not permit much competition or differential reproduction of parts from within. Species do not suppress the operation of lower levels; such effects therefore become available as exaptations conferring emergent fitness-a primary source of the different strength that species achieve as effective Darwinian individuals in evolution.


Conjugative plasmids and the degradation of arylsulfonates in Comamonas testosteroni.
Comamonas testosteroni T-2 degrades p-toluenesulfonate (TSA) via p-sulfobenzoate (PSB) and protocatechuate and degrades toluenecarboxylate via terephthalate (TER) and protocatechuate. The appropriate genes are expressed in at least five regulatory units, some of which are also found in C. testosteroni PSB-4 (F. Junker, R. Kiewitz, and A. M. Cook, J. Bacteriol. 179:919-927, 1997). C. testosteroni T-2 was found to contain two plasmids, pTSA (85 kbp) and pT2T (50 kbp); a TSA- mutant (strain TER-1) contained only plasmid pT2T. C. testosteroni PSB-4, which does not degrade TSA, contained one plasmid, pPSB (85 kbp). The type strain contained no plasmids. Conjugation experiments showed that plasmid pTSA (possibly in conjunction with pT2T) was conjugative, and the single copy of the TSA operon (tsaMBCD) with its putative regulator gene (tsaR) in strain T-2 was found on plasmid pTSA, which also carried the PSB genes (psbAC) and presumably transport for both substrates. Plasmid pTSA was assigned to the IncP1 beta group and was found to carry two copies of insertion element IS1071. Plasmid pPSB (of strain PSB-4), which could be maintained in strains with plasmid pTSA or pT2T, was also conjugative and was found to carry the PSB genes as well as to contain two copies of IS1071. In attempted conjugations with the type strain, no plasmid was recovered, but the PSB+ transconjugant carried two copies of IS1071 in the chromosome. We presume the PSB genes to be located in a composite transposon. The genes encoding the putative TER operon and degradation of protocatechuate, with the meta cleavage pathway, were attributed a chromosomal location in strains T-2 and PSB-4.


Long-term photoreceptor rescue in two rodent models of retinitis pigmentosa by adeno-associated virus delivery of Stanniocalcin-1.
Retinal degenerations, including age-related macular degeneration and the retinitis pigmentosa family of diseases, are among the leading causes of legal blindness in the United States. We previously found that Stanniocalcin-1 (STC-1) reduced photoreceptor loss in the S334ter-3 and Royal College of Surgeons rat models of retinal degeneration. The results were attributed in part to a reduction in oxidative stress. Herein, we tested the hypothesis that long-term delivery of STC-1 would provide therapeutic rescue in more chronic models of retinal degeneration. To achieve sustained delivery, we produced an adeno-associated virus (AAV) construct to express STC-1 (AAV-STC-1) under the control of a retinal ganglion cell targeting promoter human synapsin 1 (hSYN1). AAV-STC-1 was injected intravitreally into the P23H-1 and S334ter-4 rhodopsin transgenic rats at postnatal day 10. Tissues were collected at postnatal day 120 for confirmation of STC-1 overexpression and histologic and molecular analysis. Electroretinography (ERG) was performed in a cohort of animals at that time. Overexpression of STC-1 resulted in a significant preservation of photoreceptors as assessed by outer nuclear thickness in the P23H-1 (P < 0.05) and the S334ter-4 (P < 0.005) models compared to controls. Additionally, retinal function was significantly improved in the P23H-1 model with overexpressed STC-1 as assessed by ERG analysis (scotopic b-wave P < 0.005 and photopic b-wave P < 0.05). Microarray analysis identified common downstream gene expression changes that occurred in both models. Genes of interest based on their function were selected for validation by quantitative real-time PCR and were significantly increased in the S334ter-4 model.


How often do patients with psychosis fail to adhere to treatment programmes? A systematic review.
Studies that assess the rates of psychotic patients who fail to adhere to treatment programmes have generated heterogeneous results, with estimates ranging from 24 to 90%. This paper presents findings from a systematic review on adherence to treatment by patients with psychosis. Its purpose is to provide an overall estimate of treatment non-adherence in community psychiatric services, and to analyse and study patient characteristics explaining the between-study heterogeneity in rates of non-adherence. A systematic review of published studies that report rates of non-adherence with medication and scheduled appointments by psychotic patients in community settings has been undertaken. A total of 103 studies were included in this systematic review. Eighty-six of these studies were suitable for data re-analysis. The overall weighted mean rate of non-adherence, calculated in a sample of 23 796 patients, was 25.78%. A linear regression analysis of non-adherence rates on background characteristics showed that sample size was negatively associated with non-adherence rates, while first-contact cases and low-adherence cases, in comparison with ongoing cases, were associated with higher non-adherence rates. Factors associated with poor compliance included: lack of insight; positive symptoms; younger age; male gender; history of substance abuse; unemployment; and low social functioning. Approximately one in four patients with psychosis fails to adhere with treatment programmes. Preventive evidence-based clinical interventions should be routinely implemented in community settings to reduce patient non-adherence.


Evaluation of pharmacist-managed dabigatran in an inpatient setting.
Although therapeutic drug monitoring is not required for patients receiving dabigatran to determine therapeutic efficacy, there are a number of other factors to consider. Currently, there are no studies evaluating pharmacist-led management of dabigatran. The role of inpatient pharmacists related to the management of dabigatran was evaluated. All adult patients who received at least 1 dose of dabigatran with a length of stay greater than 24 hours in a single-center, community hospital between May 2011 and August 2012 were retrospectively reviewed (n = 176). Almost half of the patients (46%) required pharmacist intervention related to dabigatran management during hospital admission. Of patients receiving dabigatran prior to hospital admission, 18.4% were admitted on an inappropriate home dose. Transitioning between dabigatran and alternative anticoagulants accounted for the majority of pharmacist interventions (74.2%), with patients transitioning from unfractionated heparin to dabigatran occurring most frequently. Renal impairment requiring dose adjustment or drug discontinuation occurred in 6.3% of patients, whereas 6.8% of patients required pharmacist intervention for procedural anticoagulation. Inpatient therapy was determined to be relatively safe, with few patients (1.7%) requiring permanent discontinuation of dabigatran as a result of a bleeding complication. Pharmacists significantly contributed to the safe and appropriate use of dabigatran during hospitalization.


Effect of 7 days of bed rest on dose-response relation between plasma glucose and insulin secretion.
Physical training decreases glucose-stimulated insulin secretion. To further explore the influence of the level of daily physical activity on beta-cell secretion, the effect of 7 days of bed rest was studied in six young, healthy men by sequential hyperglycemic clamp technique (7, 11, and 20 mM glucose, each step lasting 90 min). At 11 and 20 mM glucose, insulin concentrations in plasma were higher after (87 +/- 11 and 303 +/- 63 microU/ml) than before (63 +/- 5 and 251 +/- 50 microU/ml, P less than 0.05) bed rest. Also C-peptide levels were higher after bed rest than before during glucose stimulation. The responses of other hormones, metabolites, or electrolytes influencing beta-cell secretion were not influenced by bed rest. In spite of increased insulin levels after bed rest, glucose disposal at 20 mM of glucose was significantly lower after bed rest than before. It is concluded that bed rest for 7 days increases the glucose-stimulated insulin response, at least partly due to a beta-cell adaptation increasing glucose-stimulated insulin secretion. However, the insulin secretion does not increase adequately compared with the peripheral insulin resistance induced by bed rest.


Cutaneous tuberculosis diagnosis in an inhospitable Amazonian region by means of telemedicine and molecular biology.
We report on a 13-year-old boy who displayed a chronic granulomatous inflammatory reaction of 5 years duration. The lesion was resistant to different antibiotic schemes; his routine laboratory tests and chest radiographs were normal. Teledermatologic consultation and histopathologic study of skin biopsy suggested scrofulodermal tuberculosis. Polymerase chain reaction amplification of DNA extracted from lymph node biopsy was taken as starting material for dot-blot hybridization using Mtp-40 and IS 6110 as probes for detecting either Mycobacterium tuberculosis or any mycobacteria belonging to the M tuberculosis complex, respectively. Positive results in both hybridizations were further confirmed by culturing in BACTEC MGIT 960 system. The lesion greatly diminished following isoniazid, rifampin, and ethambutol treatment. Telemedicine allowed a cutaneous tuberculosis diagnosis to be made of a patient living in a remote town located in the Amazon jungle by using molecular biology techniques.


[Incidence of urination disorders and erectile dysfunction after surgical treatment of congenital anorectal defects in children].
For assessment of the incidence and characteristics of urination disorders and erectile dysfunction after radical surgical treatment of anorectal congenital defects in children, 60 patients (12 girls, 48 boys) without urination disorders before operation, who have undergone 1-4 proctoplasties were examined. Their was Hirschsprung's disease in 20 patients, anorectal atresia in 40 patients. Iatrogenic urination disorders and erectile dysfunction were revealed in 38 (63.3%) patients, majority of them (24) underwent repeated proctoplasty. The main urination disorders were urinary incontinence (30 patients), infravesical obstruction (10), hyporeflexion of urinary bladder. Erectile dysfunction after operation was in 11 (34.4%) patients from 32. The combination of disorders was revealed in 17 (44.7%) patients. Primary radical operations in children with anorectal defects often cause iatrogenic urination disorders and erectile dysfunction. The number of complications increases progressively after repeated proctoplasty. It is necessary to improve the treatment and the methods of surgical correction bearing in mind the risk of iatrogenic disorders of genitourinary apparatus.


Laser stimulation of the chloroplast/endoplasmic reticulum nexus in tobacco transiently produces protein aggregates (boluses) within the endoplasmic reticulum and stimulates local ER remodeling.
Does the ER subdomain that associates with the chloroplast in vivo, hereafter referred to as the chloroplast/ER nexus, play a role in protein flow within the ER? In studies of tobacco cells either constitutively or transiently expressing ER-retained luminal, GFP-HDEL, or trans-membrane, YFP-RHD3, fluorescent fusion proteins, brief 405-nm (3-6-mW) laser stimulation of the nexus causes a qualitative difference in the movement and behavior of proteins in the ER. Photostimulating the nexus produces fluorescent protein punctate aggregates (boluses) within the lumen and membrane of the ER. The aggregation propagates through the membrane network throughout the cell, but within minutes can revert to normal, with disaggregation propagating back toward the originally photostimulated nexus. In the meantime, the ER grows and anastomoses around the chloroplast, forming a dense cisternal and tubular network. If this network is again photostimulated, bolus formation does not recur and, if the photostimulation results in photobleaching, fluorescence recovery after photobleaching occurs as it would typically in areas away from the nexus. Bolus propagation is not mediated by the actin cytoskeleton, but can be reversed by pre-conditioning the cells for 30 min with high, 40-45°C, temperature (heat stress). Because it is not reversed with heat stress, the reorganization of the ER at the nexus following photostimulation is a separate event.


Survival rates of patients with metastatic malignant melanoma.
Malignant melanoma (MM) is the cutaneous neoplasia with the greatest mortality rates and one of the malignancies with the highest potential of dissemination. The prognosis of patients with metastatic MM is grim, with a 5-years survival rate between 5-19%, and is dictated by the location and the number of metastases. We aimed to estimate the survival of patients with metastatic MM from our study and find out if the metastasis' location influences survival. Between 2008 and 2013, 155 patients with cutaneous MM were diagnosed in our clinic. All the patients were staged according to 2009 AJCC staging system. The median follow-up period was of 24 months. Survival was calculated by using the Kaplan-Meier method with a confidence level of 95%. 40.5% of the patients developed metastases in different organs, especially the brain. 80.6% of those with metastases died during the study. The median overall survival, estimated for the entire group of patients who developed metastases, was of 5.3 months. The influence of metastases distribution on the overall survival was examined and it was noticed that there were statistically significant differences between the risks of death of various groups of patients, depending on metastasis topography. Thus, the death probability of a patient with brain metastases is twice that of a patient with digestive metastasis, about 7 times higher than that of a patient with lung metastasis (p=0.0004) and 12 times higher than the death risk of a patient with extra-regional lymph nodes or subcutaneous metastasis (p=0.0000).


Who's at risk? Men's STD experience and condom use in southwest Nigeria.
Many STD/HIV-prevention programs worldwide assume that individuals' risk of acquiring sexually transmitted diseases, including HIV infection, is highest in the context of commercial sex. To address this assumption, research conducted in urban Southwest Nigeria combined qualitative and quantitative methods to examine men's sexual behavior, condom use, and STD experience in different types of sexual relationships (marital, casual, and commercial). Logistic regression analysis of survey data indicates that number of sexual partners and sex with sex workers are positively and significantly related to STD experience. Follow-up in-depth interviews with clients of sex workers indicate, however, that these men are actually more likely to report having contracted an STD from a casual sex partner than from a sex worker. Men are most uncertain about their vulnerability to STDs with casual partners. Men's condom use is highest in commercial sex, inconsistent in casual relationships, and lowest in marriage. STD/HIV-prevention programs need to address the range of sexual relationships and the meanings and behaviors associated with them.


Spatially constrained skin electroporation with sodium thiosulfate and urea creates transdermal microconduits.
Controlled transport of molecules through the skin's main barrier, the stratum corneum (SC), is a long standing goal of transdermal drug delivery. Traditional, needle-based injection provides delivery of almost any water soluble compound, by creating a single large aqueous pathway in the form of the hollow core of a needle, through which drug is delivered by pressure-driven flow. We extend previous work to show that SC-spanning microconduits (here with diameters of about 200 microm) can be created in vivo by skin electroporation and low-toxicity, keratolytic molecules (here sodium thiosulfate and urea). A single microconduit in isolated SC can support volumetric flow of the order of 0.01 ml s(-1) by a pressure difference of only 0.01 atm (about 10(2) Pa), demonstrating that the SC barrier has been essentially eliminated within this microscopic area.


Bispinodinium angelaceum gen. et sp. nov. (Dinophyceae), a new sand-dwelling dinoflagellate from the seafloor off Mageshima Island, Japan(1).
A new athecate dinoflagellate, Bispinodinium angelaceum N. Yamada et Horiguchi gen. et sp. nov., is described from a sand sample collected on the seafloor at a depth of 36 m off Mageshima Island, subtropical Japan. The dinoflagellate is dorsiventrally compressed and axi-symmetric along the sulcus. The morphology resembles that of the genus Amphidinium sensu lato by having a small epicone that is less than one third of the total cell length. However, it has a new type of apical groove, the path of which traces the outline of a magnifying glass. The circular component of this path forms a complete circle in the center of the epicone and the straight "handle" runs from the sulcus to the circular component. Inside the cell, a pair of elongated fibrous structure termed here the "spinoid apparatus" extends from just beneath the circular apical groove to a point near the nucleus. Each of two paired structures consists of at least 10 hyaline fibers and this is a novel structure found in dinoflagellates. Phylogenetic analyses based on the SSU and LSU RNA genes did not show any high bootstrap affinities with currently known athecate dinoflagellates. On the basis of its novel morphological features and molecular signal, we conclude that this dinoflagellate should be described as a new species belonging to a new genus.


Long-term stable vertical bone regeneration after sinus floor elevation and simultaneous implant placement with and without grafting.
Less invasive surgical approaches to regenerate bone intra-sinus and allow long-term functional implant stability are needed. To evaluate long-term vertical bone regeneration after sinus floor elevation and simultaneous implant placement with and without bone grafting. Vertical bone gains (VBG) post-sinus elevation, with and without grafting, were evaluated in thirty individuals presenting an average residual bone height (RBH) of 4.2 mm using a standardized digital technique. Measurements were taken preoperatively, and at an average of 64.6 months follow-up. Clinically, peri-implant tissues were assessed for pocket formation and presence of inflammation to evaluate established success criteria. Overall, RBH averaged 4.2 ± 1.1 mm (range: 1.8-5.8) and VBG 7.7 ± 1.6 mm (range: 6.0-12.9). Mean difference of 7.6 mm between vertical bone heights (VBH) at augmented implants sites and initial RBH, 11.8 versus 4.2 mm, (P < .0001, CI95%: 6.9-8.2) was statistically significant. RBH averaged 4.5 ± 0.8 mm and 3.8 ± 1.2 (P = .07) and VBG 6.8 ± 0.5 and 8.5 ± 1.9 mm (P = .003, CI95%: 0.6-2.7), for nongrafted and grafted individuals, respectively. The grafting group received an average graft volume of 0.35 ± 0.1 cc (range: 0.25-0.5) per implant site. Long-term follow-ups average 64.6 months (range: 36-144) and all implants met the success criteria. VBG ≥ 7 mm were 7.3 times more likely to develop on grafted sites (OR = 7.3, P = 0.02, CI95%: 1.2-46.2). None to negligible amounts of grafting material are required to regenerate substantial amounts of autogenous bone into atrophic sinus cavities after simultaneous implant placement. The regenerated VBH seems stable for functional implant stability long-term. Implant success rates were 100% at an average of 64.6 months.


[The incidence and clinical presentation of tuberculosis in HIV-positive patients].
To evaluate the features of tuberculous infection in HIV+ patients from our area we reviewed the 50 cases of seropositive patients admitted to a provincial hospital from 1985 to June 1989. TB was diagnosed in 18 of the 50 patients; mean patient age was 27 years, with a male predominance. PDA was the major risk practice. The most common localization were lymph nodes; 95% had fever and weight loss and 39% had nightly perspiration and cough; cervical lymphadenopathy and hepatosplenomegaly were found in more than 50%. Tuberculin skin test was negative in 75% of cases. The diagnosis of TB represented the diagnosis of AIDS for 66% of patients. 25 of the 50 seropositive patients met the criteria for AIDS, and 68% of these had TB. TB is a common infection in HIV+ patients from our area, and it is the first cause of the diagnosis of AIDS. The index of suspicion of TB in these patients should be high, as clinical features are often nonspecific. However, it is preventable and curable superinfection.


Mitochondrial disease in childhood: mtDNA encoded.
Since the first description of a mitochondrial DNA (mtDNA)-associated disease in the late 1980s, there have been more than 275 mutations within the mtDNA genome described causing human disease. The phenotypic expression of these disorders is vast, as disturbances of the unique physiology of mitochondria can create a wide range of clinical heterogeneity. Features of heteroplasmy, threshold effect, genetic bottleneck, mtDNA depletion, mitotic segregation, and maternal inheritance have been identified and described as a result of novel biochemical and genetic controls of mitochondrial function. We hope that as we unfold this fascinating part of clinical medicine, the reader will see how alterations in the tapestry of mitochondrial biochemistry and genetics can give rise to human illness.


Radiation-related cancer risks at low doses among atomic bomb survivors.
To clarify the information in the Radiation Effects Research Foundation data regarding cancer risks of low radiation doses, we focus on survivors with doses less than 0.5 Sv. For reasons indicated, we also restrict attention mainly to survivors within 3, 000 m of the hypocenter of the bombs. Analysis is of solid cancer incidence from 1958-1994, involving 7,000 cancer cases among 50,000 survivors in that dose and distance range. The results provide useful risk estimates for doses as low as 0.05-0.1 Sv, which are not overestimated by linear risk estimates computed from the wider dose ranges 0-2 Sv or 0-4 Sv. There is a statistically significant risk in the range 0-0.1 Sv, and an upper confidence limit on any possible threshold is computed as 0.06 Sv. It is indicated that modification of the neutron dose estimates currently under consideration would not markedly change the conclusions.


USP7 promotes non-small-cell lung cancer cell glycolysis and survival by stabilizing and activating c-Abl.
Abelson tyrosine kinase (c-Abl) is frequently mutated and highly expressed, and promotes non-small-cell lung cancer (NSCLC) survival, metastasis and tumorigenesis. c-Abl could also be modified through ubiquitination, but the underlying mechanism is not well understood. Mass spectrometry assays were performed to search c-Abl deubiquitination enzymes. The molecular mechanism was determined using Co-IP assays, pull-down assays, Western blotting upon gene knockdown or overexpression. Cell lines and animal models were used to investigate the role of c-Abl and USP7 in NSCLC. EdU staining assay and Transwell assay were performed to evaluate the proliferation and migration ability of NSCLC cells, respectively. Ubiquitin-specific protease 7 (USP7) is found to upregulate c-Abl via the deubiquitinase screen. USP7 interacts with c-Abl and decreases its K48-linked polyubiquitination, thereby increasing the stability of c-Abl. In addition to the wild-type one, c-Abl mutants can also be deubiquitinated and stabilized by USP7. Moreover, USP7 promotes c-Abl accumulation in cytoplasm by increasing its binding to 14-3-3α/β and activates the oncogenic c-Abl signalling pathway. Furthermore, the USP7/c-Abl axis promotes NSCLC cell glycolysis by direct phosphorylating and stabilizing hexokinase-2 (HK2). Knockdown of USP7 or c-Abl suppresses NSCLC cell glycolysis and reduces lactate production. Further studies revealed that overexpression of USP7 facilitates NSCLC cell growth and metastasis as well as xenograft growth in nude mice, while these activities are suppressed with USP7 or c-Abl being knocked down. USP7 is a deubiquitinase of c-Abl and upregulates its oncogenic activity. USP7 promotes NSCLC cell metabolism by activating c-Abl and HK2. Targeting the USP7/c-Abl/HK2 axis might be a potential strategy to the precision therapy of NSCLC.


[Two different outcomes after Death Cap mushroom intoxication].
Death Cap is one of the most lethal mushrooms in Denmark and may be mistaken for a non-toxic Asian mushroom. We report on two accidental cases admitted 12 and 17 hours after ingestion presenting with gastroenteritis and decline in liver function. The patient who arrived after 12 hours responded well to intensive treatment of liver failure and was discharged after 18 days. The other patient deteriorated in spite of intensive treatment and underwent liver transplantation. She was later discharged. Early diagnosis and treatment is essential.


Preoperative risk analysis in patients with adenocarcinoma or squamous cell carcinoma of the oesophagus.
The outcome after oesophagectomy depends not only on the surgical procedure itself but can be improved by the selection of patients based on a preoperative risk score. In order to compare the preoperative risk for different histological subtypes of oesophageal carcinoma, 221 patients with a squamous cell carcinoma (SCC) and 104 patients with an adenocarcinoma (AC) were enrolled in a prospective study. The preoperative assessment included socioeconomic status, alcohol and tobacco consumption, general and nutritional status, and pulmonary, cardiovascular, hepatic and renal function. More patients with an SCC were 'blue collar' and with an AC were 'white collar' workers (P < 0.001). Alcohol and tobacco consumption was higher in patients with an SCC (P < 0.01 and P < 0.05 respectively). The preoperative forced expiratory volume, arterial partial pressure of oxygen and the aminopyrine breath test showed more severely compromised pulmonary and hepatic function in patients with an SCC (P < 0.05 for all variables). Some 50 per cent of patients with an AC were overweight with a higher body mass index (P < 0.001). More patients with an AC had severely impaired cardiac function (P < 0.01). Patients with SCC and AC of the oesophagus differed regarding their socioeconomic and preoperative risk factors for surgery.


Defining the Contribution of CYP1A1 and CYP1A2 to Drug Metabolism Using Humanized CYP1A1/1A2 and Cyp1a1/Cyp1a2 Knockout Mice.
Cytochrome P450s CYP1A1 and CYP1A2 can metabolize a broad range of foreign compounds and drugs. However, these enzymes have significantly overlapping substrate specificities. To establish their relative contribution to drug metabolism in vivo, we used a combination of mice humanized for CYP1A1 and CYP1A2 together with mice nulled at the Cyp1a1 and Cyp1a2 gene loci. CYP1A2 was constitutively expressed in the liver, and both proteins were highly inducible by 2,3,7,8-tetrachlorodibenzodioxin (TCDD) in a number of tissues, including the liver, lung, kidney, and small intestine. Using the differential inhibition of the human enzymes by quinidine, we developed a method to distinguish the relative contribution of CYP1A1 or CYP1A2 in the metabolism of drugs and foreign compounds. Both enzymes made a significant contribution to the hepatic metabolism of the probe compounds 7-methoxy and 7-ehthoxyresorufin in microsomal fractions from animals treated with TCDD. This enzyme kinetic approach allows modeling of the CYP1A1, CYP1A2, and non-CYP1A contribution to the metabolism of any substrate at any substrate, inhibitor, or enzyme concentration and, as a consequence, can be integrated into a physiologically based pharmacokinetics model. The validity of the model can then be tested in humanized mice in vivo. SIGNIFICANCE STATEMENT: Human CYP1A1 and CYP1A2 are important in defining the efficacy and toxicity/carcinogenicity of drugs and foreign compounds. In light of differences in substrate specificity and sensitivity to inhibitors, it is of central importance to understand their relative role in foreign compound metabolism. To address this issue, we have generated mice humanized or nulled at the Cyp1a gene locus and, through the use of these mouse lines and selective inhibitors, developed an enzyme kinetic-based model to enable more accurate prediction of the fate of new chemicals in humans and which can be validated in vivo using mice humanized for cytochrome P450-mediated metabolism.


Abnormal alpha-synuclein interactions with rab3a and rabphilin in diffuse Lewy body disease.
The present study examines alpha-synuclein interactions with rab3a and rabphilin by antibody arrays, immunoprecipitation and pull-down methods in the entorhinal cortex of control cases and in diffuse Lewy body disease (LBD) cases. Alpha-synuclein immunoprecipitation revealed alpha-synuclein binding to rabphilin in control but not in LB cases. Immunoprecipitation with rab3a disclosed rab3a binding to rabphilin in control but not in LB cases. Moreover, rab3a interacted with high molecular weight (66 kDa) alpha-synuclein only in LB cases, in agreement with parallel studies using antibody arrays. Results were compared with pull-down assays using His(6)/Flag-tagged rab3, rab5 and rab8, and anti-Flag immunoblotting. Weak bands of 17 kDa, corresponding to alpha-synuclein, were obtained in LB and, less intensely, in control cases. In addition, alpha-synuclein-immunoreactive bands of high molecular weight (36 kDa) were seen only in LB cases after pull-down assays with rab3a, rab5 or rab8. These findings corroborate previous observations showing rab3a-rabphilin interactions in control brains, and add substantial information regarding decreased binding of rab3a to rabphilin and increased binding of rab3a to alpha-synuclein aggregates in LB cases. Since, alpha-synuclein, rab3a and rabphilin participate in the docking and fusion of synaptic vesicles, it can be suggested that exocytosis of neurotransmitters may be impaired in LB diseases.


The Added Value of Log File Analyses of the Use of a Personal Health Record for Patients With Type 2 Diabetes Mellitus: Preliminary Results.
The electronic personal health record (PHR) is a promising technology for improving the quality of chronic disease management. Until now, evaluations of such systems have provided only little insight into why a particular outcome occurred. The aim of this study is to gain insight into the navigation process (what functionalities are used, and in what sequence) of e-Vita, a PHR for patients with type 2 diabetes mellitus (T2DM), to increase the efficiency of the system and improve the long-term adherence. Log data of the first visits in the first 6 weeks after the release of a renewed version of e-Vita were analyzed to identify the usage patterns that emerge when users explore a new application. After receiving the invitation, 28% of all registered users visited e-Vita. In total, 70 unique usage patterns could be identified. When users visited the education service first, 93% of all users ended their session. Most users visited either 1 or 5 or more services during their first session, but the distribution of the routes was diffuse. In conclusion, log file analyses can provide valuable prompts for improving the system design of a PHR. In this way, the match between the system and its users and the long-term adherence has the potential to increase.


Improving pediatric asthma control among minority children participating in medicaid: providing practice redesign support to deliver a chronic care model.
Asthma, a leading chronic disease of children, currently affects about 6.2 million (8.5%) children in the United States. Despite advances in asthma research and availability of increasingly effective therapy, many children do not receive appropriate medications to control the disease, have over-reliance on reliever medication, and lack systematic follow-up care. The situation is even worse for poor inner-city and minority children who have significantly worse asthma rates, severity, and outcomes. National Asthma Education and Prevention Program Guidelines recommend a multimodal, chronic care approach. The authors assessed the effectiveness of practice redesign and computerized provider feedback in improving both practitioner adherence to National Asthma Education and Prevention Program Guidelines (NAEPP), and patient outcomes in 295 poor minority children across four Federally Qualified Health Centers (FQHC). In a nonrandomized, two-group (intervention versus comparison), two-phase trial, all sites were provided redesign support to provide quarterly well-asthma visits using structured visit forms, community health workers for outreach and follow-up, a Web-based disease registry for tracking and scheduling, and a provider education package. Intervention sites were given an additional Web-based, computerized patient-specific provider feedback system that produced a guideline-driven medication assessment prompt. Logistic regression results showed that providers at intervention sites were more than twice as likely on average to prescribe guideline-appropriate medications after exposure to our feedback system during the Phase I enrollment period than providers at comparison sites (exp(B) = 2.351, confidence interval [CI] = 1.315-4.204). In Phase II (the post-enrollment visit period), hierarchical linear models (HLMs) and latent growth curves were used to show that asthma control improved significantly by .19 (SE = .05) on average for each of the remaining four visits (about 11% of a standard deviation), and improved even more for patients at intervention sites. These results show that implementation of practice redesign support guided by a pediatric chronic care model can improve provider adherence to treatment guidelines as well as patients' asthma control. The addition of patient-specific feedback for providers results in quicker adoption of guideline recommendations and potentially greater improvements in asthma control compared to the basic practice redesign support alone.


LIPID MAPS: update to databases and tools for the lipidomics community.
LIPID MAPS (LIPID Metabolites and Pathways Strategy), www.lipidmaps.org, provides a systematic and standardized approach to organizing lipid structural and biochemical data. Founded 20 years ago, the LIPID MAPS nomenclature and classification has become the accepted community standard. LIPID MAPS provides databases for cataloging and identifying lipids at varying levels of characterization in addition to numerous software tools and educational resources, and became an ELIXIR-UK data resource in 2020. This paper describes the expansion of existing databases in LIPID MAPS, including richer metadata with literature provenance, taxonomic data and improved interoperability to facilitate FAIR compliance. A joint project funded by ELIXIR-UK, in collaboration with WikiPathways, curates and hosts pathway data, and annotates lipids in the context of their biochemical pathways. Updated features of the search infrastructure are described along with implementation of programmatic access via API and SPARQL. New lipid-specific databases have been developed and provision of lipidomics tools to the community has been updated. Training and engagement have been expanded with webinars, podcasts and an online training school.


[Gross Hematuria after kidney biopsy. A case report].
We describe the clinical case of a patient experiencing severe gross hematuria causing clotting in the renal pelvis, after undergoing a kidney biopsy.The ecocolordopper and CT angiography performed did not reveal the cause of hematuria.The kidney arteriography allowed the diagnosis, revealing an arteriovenous fistula responsible for bleeding together with a small false aneurysm in the lower pole of the biopsied kidney. Both lesions were successfully treated with superselective embolization with microcoils. We discuss about the diagnostic and therapeutic approach of these rare post-biopsy complications briefly focusing on the technical aspects and on possible risks that the transcatheter embolotherapy may result.


Signaling in morphogenesis: transport cues in morphogenesis.
Extracellular transport processes play critical roles in morphogenesis. While diffusive transport effects on morphogenesis are well illustrated in examples like blood capillary architecture and in cell morphogenetic responses to the local extracellular protein environment, the effects of fluid convection, although important in many developing and regenerating tissues, are not well understood. Convective forces are present whenever a hydrated tissue undergoes dynamic mechanical strain, and so convection could not only dominate the transport of large molecules like proteins, but might also serve as a mechanism for mechanosensing. The complex interdependence of mechanical forces, protein transport and extracellular morphogen gradients needs to be elucidated in a comprehensive way in order for the importance of transport on morphogenesis to be fully appreciated.


Effect of experience of severe stroke on subjective valuations of quality of life after stroke.
Previous work suggests that the quality of life associated with severe disability after stroke is rated very poorly by members of the public, often as being worse than death. Other evidence suggests that experience of illness alters perceptions of its severity. This was tested for severe stroke. Eleven patients with severely disabling stroke, but able to complete a standard gamble interview, 22 age and sex matched controls, and 20 health professionals participated. A standard gamble interview was carried out to determine the quality of life (utility) associated with three hypothetical scenarios representing mild, moderate, and severe stroke, and current health. A sample was retested for reliability, and comparisons were made with other measures of health status. All three subject groups showed wide variation in the utilities they attached to each of the scenarios. The control subjects' valuations were lower than those of either patients or staff members, especially for moderate stroke (median 0.30, 0.73, and 0.68 respectively). There were weak to moderate correlations between utilities and other measures of health status including the Barthel index (r=0.51) and Rivermead mobility score (r=0.24). Test retest-reliability was modest (reliability coefficient 0.75), but indicators of the internal validity of the results were good. In conclusion, it cannot be assumed that general population valuations are valid for patient groups. In clinical practice it is unsafe to make any assumption about subjective quality of life after stroke, due to the wide range of valuations given, although many people rate severe and moderate stroke at least as bad as death.


The challenging management of pediatric corneal transplantation: an overview of surgical and clinical experiences.
Pediatric keratoplasty is an immense challenge because of the technical complexity of the procedure and the high risk of rejection in young graft recipients. Our aim is to describe the major indications and appropriate timing of corneal transplantation intervention, including a report of our experience and surgical tips, in conjunction with the current literature. Review of the literature on indications for keratoplasty in pediatric patients classified as: congenital, traumatic and acquired non-traumatic opacities. We additionally explored the challenges corneal surgeons face in performing this type of surgery and review the most pressing transplant-related problems and their management. Outcomes after pediatric keratoplasty, in terms of visual development, restoration and clarity of the graft, are influenced by peri-operative local and systemic conditions and factors, and by intraoperative management of the transplantation procedure itself. Pediatric corneal transplantation is a critical tool for visual restoration and development in young patients with corneal opacities, particularly during the critical period of visual development. Successful management of the significant challenges associated with pediatric keratoplasty requires customized clinical and surgical management of each patient with particular attention paid to proper post-operative rehabilitation.


The development of an ecological approach to manage the pollution risk from highway runoff.
In the UK, the Highways Agency is responsible for operating, maintaining and improving the strategic road network in England. One focus of the Highways Agency's ongoing research into the nature and impact of highway runoff is aimed at ensuring that the Highways Agency will meet the requirements of the EU Water Framework Directive. A research programme, undertaken in partnership with the Environment Agency, is in progress to develop a better understanding of pollutants in highway runoff and their ecological impact. The paper presents the outcome of a study to: (1) monitor pollutants in highway runoff under different climate and traffic conditions; (2) develop standards to assess potential ecological risks from soluble pollutants in highway runoff; and (3) develop a model to predict pollutant concentrations in highway runoff. The model has been embedded in a design tool incorporating risk assessment procedures and receiving water standards for soluble and insoluble pollutants--the latter has been developed elsewhere in another project within the research programme. The design tool will be used to support improved guidance on where, and to what level, treatment of runoff is required for highway designers to manage the risk of ecological impact from highway runoff.


Changes in immunoreactive beta-endorphin, methionine-enkephalin and ACTH in bone marrow cells and fluid from leukemic children.
The present study demonstrates the presence of the endogenous opioid peptides, beta-endorphin (beta-EP) and methionine-enkephalin (MET-ENK), and of ACTH in cell homogenates and interstitial fluid from sternal biopsy of leukemic children. The peptides were identified by chromatography and radioimmunoassay. In leukemic children with lymphoblastic cells present in the sternal sample, concentrations of immunoreactive (ir) beta-EP in the cell homogenate, but not in the fluid, were significantly higher than in leukemic children with normal bone marrow. In contrast, ir MET-ENK and ir ACTH did not differ between the two study groups either in the cell homogenate or in the fluid. These data suggest the presence of a complex system of opioid peptides in the cells and interstitial fluid of bone marrow of leukemic children with the highest concentrations of ir beta-EP appearing in samples collected during the active phase of the disease, and may suggest a possible role of opioid peptides as immunomodulatory substances.


Hemostatic Management of Trauma-Induced Coagulopathy.
Trauma-induced coagulopathy is a primary factor in many trauma-related fatalities. Management hinges upon rapid diagnosis of coagulation abnormalities and immediate administration of appropriate hemostatic agents. Use of crystalloids and packed red blood cells has traditionally been the core of trauma resuscitation, but current massive transfusion protocols include combination therapy with fresh frozen plasma and predefined ratios of platelets to packed red blood cells, limiting crystalloid administration. Hemostatic agents such as tranexamic acid, prothrombin complex concentrate, fibrinogen concentrate, and, in cases of refractory bleeding, recombinant activated factor VIIa may also be warranted. Goal-directed resuscitation using viscoelastic tools allows specific component-centered therapy based on individual clotting abnormalities that may limit blood product use and thromboembolic risks and may lead to reduced mortality. Because of the complex management of patients with trauma-induced coagulopathy, critical care nurses must be familiar with the pathophysiology, acute diagnostics, and pharmacotherapeutic options used to treat these patients.


Chemical Recycling of Polyethylene by Tandem Catalytic Conversion to Propylene.
Although polyethylene (PE) and polypropylene (PP) are by far the world's largest volume plastics, only a tiny fraction of these energy-rich polyolefins are currently recycled. Depolymerization of PE to its constituent monomer, ethylene, is highly endothermic and conventionally accessible only through unselective, high-temperature pyrolysis. Here, we provide experimental demonstrations of our recently proposed tandem catalysis strategy, which uses ethylene to convert PE to propylene, the commodity monomer used to make PP. The approach combines rapid olefin metathesis with rate-limiting isomerization. Monounsaturated PE is progressively disassembled at modest temperatures via many consecutive ethenolysis events, resulting selectively in propylene. Fully saturated PE can be converted to unsaturated PE starting with a single transfer dehydrogenation to ethylene, which produces a small amount of ethane (1 equiv per dehydrogenation event). These principles are demonstrated using both homogeneous and heterogeneous catalysts. While selectivity under batch conditions is limited at high conversion by the formation of an equilibrium mixture of olefins, high selectivity to propylene (≥94%) is achieved in a semicontinuous process due to the continuous removal of propylene from the reaction mixture.


Strategies to Identify Patient Risks of Prescription Opioid Addiction When Initiating Opioids for Pain: A Systematic Review.
Although prescription opioid use disorder is associated with substantial harms, strategies to identify patients with pain among whom prescription opioids can be safely prescribed have not been systematically reviewed. To review the evidence examining factors associated with opioid addiction and screening tools for identifying adult patients at high vs low risk of developing symptoms of prescription opioid addiction when initiating prescription opioids for pain. MEDLINE and Embase (January 1946 to November 2018) were searched for articles investigating risks of prescription opioid addiction. Original studies that were included compared symptoms, signs, risk factors, and screening tools among patients who developed prescription opioid addiction and those who did not. Two investigators independently assessed quality to exclude biased or unreliable study designs and extracted data from higher quality studies. The Preferred Reporting Items for Systematic Reviews and Meta-analyses of Diagnostic Accuracy Studies (PRISMA-DTA) reporting guideline was followed. Likelihood ratios (LRs) for risk factors and screening tools were calculated. Of 1287 identified studies, 6 high-quality studies were included in the qualitative synthesis and 4 were included in the quantitative synthesis. The 4 high-quality studies included in the quantitative synthesis were all retrospective studies including a total of 2 888 346 patients with 4470 cases that met the authors' definitions of prescription opioid addiction. A history of opioid use disorder (LR range, 17-22) or other substance use disorder (LR range, 4.2-17), certain mental health diagnoses (eg, personality disorder: LR, 27; 95% CI, 18-41), and concomitant prescription of certain psychiatric medications (eg, atypical antipsychotics: LR, 17; 95% CI, 15-18) appeared useful for identifying patients at high risk of opioid addiction. Among individual findings, only the absence of a mood disorder (negative LR, 0.50; 95% CI, 0.45-0.52) was associated with a lower risk of opioid addiction. Despite their widespread use, most screening tools involving combinations of questions were based on low-quality studies or, when diagnostic performance was assessed among high-quality studies, demonstrated poor performance in helping to identify patients at high vs low risk. While a history of substance use disorder, certain mental health diagnoses, and concomitant prescription of certain psychiatric medications appeared useful for identifying patients at higher risk, few quality studies were available and no symptoms, signs, or screening tools were particularly useful for identifying those at lower risk.


Managing Advanced HIV Disease in a Public Health Approach.
In 2017, the World Health Organization (WHO) published guidelines for the management of advanced human immunodeficiency virus (HIV) disease within a public health approach. Recent data suggest that more than a third of people starting antiretroviral therapy (ART) do so with advanced HIV disease, and an increasing number of patients re-present to care at an advanced stage of HIV disease following a period of disengagement from care. These guidelines recommend a standardized package of care for adults, adolescents, and children, based on the leading causes of morbidity and mortality: tuberculosis, severe bacterial infections, cryptococcal meningitis, toxoplasmosis, and Pneumocystis jirovecii pneumonia. A package of targeted interventions to reduce mortality and morbidity was recommended, based on results of 2 recent randomized trials that both showed a mortality reduction associated with delivery of a simplified intervention package. Taking these results and existing recommendations into consideration, WHO recommends that a package of care be offered to those presenting with advanced HIV disease; depending on age and CD4 cell count, the package may include opportunistic infection screening and prophylaxis, including fluconazole preemptive therapy for those who are cryptococcal antigen positive and without evidence of meningitis. Rapid ART initiation and intensified adherence interventions should also be proposed to everyone presenting with advanced HIV disease.


Heartwater. The morphology of Cowdria ruminantium and its staining characteristics in the vertebrate host and in vitro.
The morphology of Cowdria ruminantium is described and its staining characteristics in the vertebrate host and in vitro are summarized. Morphologically, the organisms are characterized in the cytoplasm of endothelial cells, macrophages and reticulo-endothelial cells. Based on the morphology of the internal structure of the organisms, elementary (electron-dense), intermediate and reticulate bodies are identified. Each organism is surrounded by a double membrane and a "capsule" is evident around a few organisms in vitro. Usually, only organisms of the same form are found within a particular vacuole, although mixed colonies are described in the in vitro studies.


Propofol modulates phasic and tonic GABAergic currents in spinal ventral horn interneurones.
Surgical interventions like skin incisions trigger withdrawal reflexes which require motor neurones and local circuit interneurones in the spinal ventral horn. This region plays a key role in mediating immobilizing properties of the GABAergic anaesthetic propofol. However, it is unclear how propofol modulates GABA(A) receptors in the spinal ventral horn and whether tonic or phasic inhibition is involved. Organotypic spinal cord tissue slices were prepared from mice. Whole-cell recordings were performed for quantifying effects of propofol on GABA(A) receptor-mediated phasic transmission and tonic conductance. Propofol increased GABAergic phasic transmission by a prolongation of the decay time constant in a concentration-dependent manner. The amount of the charge transferred per inhibitory post-synaptic current, described by the area under the curve, was significantly augmented by 1 µM propofol (P<0.01). A GABA(A) receptor-mediated tonic current was not induced by 1 µM propofol but at a concentration of 5 µM (P<0.05). Propofol depresses ventral horn interneurones predominantly by phasic rather than by tonic GABA(A) receptor-mediated inhibition. However, the present results suggest that the involvement of a tonic inhibition might contribute to the efficacy of propofol to depress nociceptive reflexes at high concentrations of the anaesthetic.


Endocytic Roles of Glycans on Proteins and Lipids.
Most cell surface proteins are decorated by glycans, and the plasma membrane is rich in glycosylated lipids. The mechanisms by which the enormous complexity of these glycan structures on proteins and lipids is exploited to control glycoprotein activity by setting their cell surface residence time and the ways by which they are taken up into cells are still under active investigation. Here, two mechanisms are presented, termed galectin lattices and glycolipid-lectin (GL-Lect)-driven endocytosis, which are among the most prominent to establish a link between glycan information and endocytosis. Types of glycans on glycoproteins and glycolipids are reviewed from the angle of their interaction with glycan-binding proteins that are at the heart of galectin lattices and GL-Lect-driven endocytosis. Examples are given to show how these mechanisms affect cellular functions ranging from cell migration and signaling to vascularization and immune modulation. Finally, outstanding challenges on the link between glycosylation and endocytosis are discussed.


Subtle Changes in the Combining Site of the Chlamydiaceae-Specific mAb S25-23 Increase the Antibody-Carbohydrate Binding Affinity by an Order of Magnitude.
Murine antibodies S25-23, S25-26, and S25-5 derive from a common germ-line origin, and all bind the Chlamydiaceae family-specific epitope αKdo(2→8)αKdo(2→4)αKdo (where Kdo is 3-deoxy-α-d- manno-oct-2-ulosonic acid) with high affinity and specificity. These antibodies recognize the entire trisaccharide antigen in a linkage-dependent manner via a groove composed largely of germ-line residues. Despite sharing identical heavy and light chain genes, S25-23 binds the family-specific epitope with nanomolar affinity, which is an order of magnitude higher than that of S25-26, while S25-5 displays an affinity between those of S25-23 and S25-26. We determined the high-resolution crystal structures of S25-23 and S25-5 antigen binding fragments in complex with a pentasaccharide derived from the LPS of Chlamydia and measured the affinity of S25-5 for chlamydial LPS antigens using isothermal titration microcalorimetry. The 1.75 Å resolution structure of S25-23 shows how subtle conservative mutations Arg(L)-27E to lysine and Ser(H)-56 to threonine lead to an order of magnitude increase in affinity. Importantly, comparison between previous S25-26 structures and the 1.99 and 2.05 Å resolution liganded and unliganded structures of S25-5, respectively, shows how a Ser(L)-27E mutation results in an intermediate affinity due to the reduced enthalpic penalty associated with complex formation that would otherwise be required for arginine in this position. This strategy allows for subtle adjustments in the combining site via affinity maturation that have dramatic consequences for the affinity of an antibody for its antigen.


Determining the parameters of concentration-response relationship for histamine on the isolated portion of guinea pig stomach and guinea pig right atrium.
In our previous experiments measuring acid secretion in the isolated mouse stomach we found higher EC50 values (agonist concentration, evoking half maximal effect) for histamine than in the experiments on the isolated guinea pig right atrium (contraction frequency). The difference could be due to nonequilibrium conditions in the isolated mouse stomach preparation or to species differences. In order to check this possibility and to test the consistency of concentration-response relationship (CRR) parameter determination on an isolated secretory organ, we compared EC50 for histamine and pA2 for famotidine on the two isolated preparations from the same species, guinea pigs. Both, EC50 and pA2 values were significantly different when measuring gastric acid secretion (9.7 microM and 6.07) from the parameters, found in the isolated right atrium measuring frequency of contractions (1.6 microM and 7.06). From these results it can be concluded that the isolated portion of guinea pig stomach is suitable for CRR determination; but the parameters obtained are in the range of higher concentrations than the real ones because of kinetic particularities of the preparation.


Vasculitis of the upper airways.
Systemic vasculitides are rare and potentially life-threatening diseases. Granulomatosis with polyangiitis (GPA; formerly Wegener's granulomatosis) and Churg Strauss syndrome (possibly to be renamed eosinophilic granulomatosis with polyangiitis; EGPA) are the 2 chief systemic vasculitides which may involve the upper respiratory tract. Chronic allergic rhinitis and nasal polyposis in EGPA, and recurrent sinusitis and/or otitis in both conditions, are not specific and can thus represent real diagnostic challenges if they are the first manifestations of the disease. Nasal septum perforation, saddle nose deformity and/or subglottic stenosis (SGS), although not totally specific, are more suggestive of GPA. Because upper airway manifestations often tend to be refractory to systemic therapy and/or to linger, local treatment represents a major aspect of management of the condition, especially for patients with SGS.


Consolidation of medical groups into physician practice management organizations.
Medical groups are growing and merging to improve efficiency and bargaining leverage in the competitive managed care environment. An increasing number are affiliating with physician practice management (PPM) firms that offer capital financing, expertise in utilization management, and global capitation contracts with health insurance entities. These physician organizations provide an alternative to affiliation with a hospital system and to individual physician contracting with health plans. To describe the growth, structure, and strategy of PPM organizations that coordinate medical groups in multiple markets and contract with health maintenance organizations (HMOs). Case studies, including interviews with administrative and clinical leaders, review of company documents, and analysis of documents from investment bankers, the Securities and Exchange Commission, and industry observers. Medical groups and independent practice associations (IPAs) in California and New Jersey affiliated with MedPartners, FPA Medical Management, and UniMed. Growth in number of primary care and specialty care physicians employed by and contracting with affiliated medical groups; growth in patient enrollment from commercial, Medicare, and Medicaid HMOs; growth in capitation and noncapitation revenues; structure and governance of affiliated management service organizations and professional corporations; and contracting strategies with HMOs. Between 1994 and 1996, medical groups and IPAs affiliated with 3 PPMs grew from 3787 to 25763 physicians; 65% of employed physicians provide primary care, while the majority of contracting physicians provide specialty care. Patient enrollment in HMOs grew from 285503 to 3028881. Annual capitation revenues grew from $190 million to $2.1 billion. Medical groups affiliated with PPMs are capitated for most professional, hospital, and ancillary clinical services and are increasingly delegated responsibility by HMOs for utilization management and quality assurance. Physician practice management organizations and their affiliated medical groups face the challenge of continuing rapid growth, sustaining stock values, and improving practice efficiencies while maintaining the loyalty of physicians and patients.


Characterization of circulating and endothelial progenitor cells in patients with extreme-duration type 1 diabetes.
We characterized and correlated endothelial progenitor cells (EPCs) and circulating progenitor cells (CPCs) with lack of vascular complications in the Joslin Medalist Study in patients with type 1 diabetes for 50 years or longer. EPC and CPC levels were ascertained by flow cytometry and compared among Medalists (n = 172) with or without diabetic retinopathy (DR; n = 84 of 162), neuropathy (n = 94 of 165), diabetic nephropathy (DN; n = 18 of 172), cardiovascular disease (CVD; n = 63 of 168), age-matched controls (n = 83), type 2 diabetic patients (n = 36), and younger type 1 diabetic patients (n = 31). Mitogens, inflammatory cytokines, and oxidative markers were measured in blood or urine. Migration of cultured peripheral blood mononuclear cells (PBMCs) from Medalists and age-matched controls were compared. Medalists' EPC and CPC levels equaled those of their nondiabetic age-matched controls, were 10% higher than those in younger type 1 diabetic patients, and were 20% higher than those in age-matched type 2 diabetic patients. CPC levels were 15% higher in Medalists without CVD and nephropathy than in those affected, whereas EPC levels were significantly higher in those without peripheral vascular disease (PVD) than those with PVD. Stromal-derived factor 1 (SDF-1) levels were higher in Medalists with CVD, DN, and DR than in those not affected and their controls. IGF-I levels were lower in Medalists and correlated inversely with CPC levels. Additionally, cultured PBMCs from Medalists migrated more than those from nondiabetic controls. Normal levels of EPC and CPC in the Medalists, unlike other groups with diabetes, especially those without CVD, support the idea that endogenous factors exist to neutralize the adverse effects of metabolic abnormalities of diabetes on vascular tissues.


COVID-19 and trained immunity: the inflammatory burden of long covid.
Severe COVID-19 elicits excessive inflammation mediated by innate immune cells like monocytes. Recent evidence reveals extensive epigenetic changes in monocytes during recovery from severe COVID-19, including increased chromatin accessibility at genes related to cytokine production and leukocyte activation. These changes likely originate from the reprogramming of upstream hematopoietic stem and progenitor cells (HSPCs) and represent "trained immunity". HSPC-to-monocyte transmission of epigenetic memory may explain the persistence of these monocyte alterations despite their short lifespan. IL-6 appears pivotal for imprinting durable epigenetic modifications in monocytes during acute infection, with IL-1β potentially playing a contributory role. The poised inflammatory phenotype of monocytes post-COVID-19 may drive chronic inflammation and tissue damage, contributing to post-acute sequelae of COVID-19 symptoms. COVID-19 could also exacerbate inflammation-related diseases, such multisystem inflammatory syndromes, by altering innate immune tendencies via hematopoietic epigenetic reprogramming. Further clinical investigations quantifying inflammatory mediators and mapping epigenetic changes in HSPCs/monocytes of recovering patients are warranted. Research should also examine whether COVID-19 elicits transgenerational inheritance of epigenetic alterations. Elucidating mechanisms underlying COVID-19-induced monocyte reprogramming and developing interventions targeting key inflammatory regulators like IL-6 may mitigate the sustained inflammatory burden imposed by the aberrant trained immunity post-COVID-19.


[Relationship of social support and meaning of life to suicidal thoughts in cancer patients].
This study was done to explore the relationship of social support and meaning of life to suicidal thoughts among patients with cancer. Data were collected by questionnaires from 138 patients who had been in cancer treatment at medical clinics and 8 patients who were members of an internet cancer association. The data were collected between August and November 2009 and analyzed using SPSS 12.0. Of the participants, 47.3% reported having had suicidal thoughts and 16.4% had attempted suicide since the diagnosis of cancer. The study participants received most support from family members, but 73.3% reported experiencing an existential vacuum. The suicide attempt group had significantly higher scores according to gender, age, level of education, diagnosis, treatment modality, level of activity, caregiver and social support compare to the suicide thought group. Suicidal thoughts were negatively related to social support and meaning of life was positively associated with social support. Support from family and friends and diagnosis explained 50.0% of variance for suicidal thoughts with 36.0% of variance being explained by family support. Nurses should be able to identify risk factors for suicide in cancer patients. Prevention and intervention efforts need to be directed toward improving social support, family support in particular, and assisting patients finding meaning in life after a diagnosis of cancer.


A new genus and species of the family rhabdolaimidae (nematoda), with descriptions of two known species and taxonomic discussion.
A new genus, one new and two known species belonging to family Rhabdolaimidae are described and illustrated. Mediolaimus n. gen. is characterized by small-sized body; papilliform outer labial and cephalic sensilla; long, tubular stoma with stegostom having relatively larger dorsal and smaller sub ventral denticles at same level; cylindrical pharynx with expanded basal bulb having thickened lumen; didelphic gonad having outstretched ovaries; elongate-conoid tail with hemispherical terminus without mucro or spinneret and caudal glands obscure. Mediolaimus obtusicaudatus n. gen. n. sp. is characterized by stoma with a relatively larger dorsal denticle and two fine subventral denticles at same level; amphidelphic reproductive system with outstretched ovaries; tail elongate conoid with smooth hemispherical terminus and absence of male. Rhabdolaimus terrestrisde Man, 1880 and Udonchus tenuicaudatusCobb, 1913 are described with additional details. The inter population variations are discussed in the latter species along with a detailed note on the status of Rhabdolaimidae and its proposed placement under superfamily Rhabdolaimoidea of suborder Campydorina.


Mind wandering in anxiety disorders: A status report.
Many investigations have targeted the subject of worry in anxiety disorders. Worry can be regarded as a subtype of mind wandering (MW), which is undeliberate, perseverative, negatively-valenced, and mainly future-oriented. Nevertheless, until now only a few studies have explored the role of overall MW in the origin and course of anxiety disorders. To foster progress in this field, we briefly describe and discuss relevant studies addressing MW in subjects with anxiety disorders or symptoms or disorders associated with anxiety symptoms. Provisional synthesis suggests that: a) the overall amount of MW is positively correlated with anxiety symptoms; b) MW characteristics reflecting worry and rumination appear to be relevant in anxiety; c) comorbid depressive and ADHD symptoms may contribute to excessive MW in anxiety; d) MW-related therapeutic interventions may be useful as complementary treatments in anxiety disorders. However, more studies related to MW in anxiety disorders or symptoms are necessary to corroborate and extend these initial findings. Such investigations should ideally combine experience sampling with self-rating assessments of both MW and worry/rumination.


Ovariectomy increases RANKL protein expression in bone marrow adipocytes of C3H/HeJ mice.
Estrogen deficiency induces bone loss by increasing bone resorption, in part through upregulation of receptor activator of nuclear factor-κB ligand (RANKL). RANKL is secreted by osteoblasts and osteocytes, but more recently bone marrow (pre)adipocytes have also been shown to express RANKL. Estrogen deficiency increases bone marrow adipose tissue (BMAT). The aim of this study was to determine the effect of ovariectomy (OVX) on RANKL protein expression by bone marrow adipocytes in C3H/HeJ mice. Fourteen-week-old female C3H/HeJ mice (n = 20) were randomized to sham surgery (Sham) or OVX. After 4 wk animals were euthanized. BMAT volume fraction (BMAT volume/marrow volume) was quantified by polyoxometalate-based contrast-enhanced nano-computed tomography. The percentage of RANKL-positive bone marrow adipocytes (RANKL-positive bone marrow adipocytes/total adipocytes) and the percentage of RANKL-positive osteoblasts covering the bone surface (bone surface covered in RANKL-positive osteoblasts/total bone surface) were quantified in the distal metaphysis of immunohistochemically stained sections of the left femur. The effects of OVX were analyzed by Student's t test or Mann-Whitney U test. RANKL was detected in osteoblasts, osteocytes, and bone marrow adipocytes. OVX significantly increased mean percentage of RANKL-positive bone marrow adipocytes [mean (SD): Sham 42 (18)%; OVX 64 (12)%; P = 0.029] as well as BMAT volume/marrow volume [median (interquartile range): Sham 1.4 (4.9)%; OVX 7.2 (7.3)%; P = 0.008] compared with Sham. We show that OVX increased both the percentage of RANKL-positive bone marrow adipocytes and the total BMAT volume fraction in C3H/HeJ mice. Therefore, RANKL produced by bone marrow adipocytes could be an important contributor to OVX-induced bone loss in C3H/HeJ mice.


Alpha 1 blockers: antihypertensives whose positive metabolic profile with regard to hyperinsulinaemia and lipid metabolism cannot be ignored.
The selective alpha 1-adrenoceptor-blocking agents constitute effective monotherapy in the control of essential hypertension. The failure of well-controlled trials of other antihypertensives to achieve the expected reduction in coronary heart disease has given rise to speculation that the antihypertensives used are atherogenic because of metabolic changes induced in the patient. The selective alpha 1-adrenoceptor blockers have positive effects on carbohydrate metabolism, hyperinsulinaemia and lipid metabolism. These changes, which consist of a modest reduction in total serum cholesterol, an increase in high-density-lipoprotein cholesterol, and a decrease in low-density lipoprotein, very-low-density lipoprotein and triglyceride, accompanied by a reduction in hyperglycaemia and serum insulin levels, have been observed in hypertensive patients. Experimental studies using isolated tissue culture and intact animal systems have confirmed these observations. The changes in metabolism may represent the key to prevention of coronary heart disease in the hypertensive population.


N-Hydroxylation of 1-(2,5-demethoxy-4-methylphenyl)-2-aminopropane by rabbit liver microsomes.
Metabolic N-hydroxylation of the potent psychotomimetic amine 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane (5) by rabbit liver microsomal preparations has been investigated. Synthetic hydroxylamine 8 was obtained by sequential reduction of the corresponding nitropropene 10 with sodium borohydride followed by zinc reduction of the resulting nitropropane 11. Compound 8 in water (pH 7.4) was rapidly air oxidized to oxime 12; this oxidation was completely blocked by rabbit liver microsomes. Microsomal incubations of amine 5 or its bis(methoxy-d3)hexadeuterio analog 5-d6 resulted in the formation of 8 and 8-d6, respectively, identified as their bis(trifluoroacetyl) derivatives by GLC-MS. Quantitative estimations of metabolite formation employing selected ion monitoring with the aid of an accelerating voltage alternator were accomplished by stable isotope dilution analyses with 5-d6 as substrate and 8-d0 as internal standard. Similar analyses starting with "pseudoracemates" (R)-5-d0:(S)-5-d6 or (R)-5-d6:(S)-5-d0 as substrates established metabolite 8 to be enriched with its R enantiomer.


[Method of dynamic dosage fractionation in preoperative radiation therapy of esophageal cancer].
Dynamic dose fractionation in preoperative radiation therapy in 53 esophageal cancer patients comes down to the following: in the first 3 days a daily tumor dose was 4 Gy, then a daily dose was 2 Gy with a summary focal dose up to 36-42 Gy. Therapeutic results for 127 patients served as control using the routine fractionation schedule: 2 Gy daily (5 times a week) up to a summary focal dose of 40-45 Gy. The irradiated area included almost the entire length of the organ with the regional lymphatic collector. Good tolerance of the method of dynamic dose fractionation was noted: no increased frequency and severity of radiation reactions and complications were noted. The use of the new method made it possible to reduce the periods of preoperative irradiation and rehabilitation. Considerable tumor regression after the use of the above method was observed in 69 +/- 8.7% of the patients, in the control group in 38.9 +/- 8.2% only. A simultaneous increase in the degree of radiation injury of tumors was noted in the study group (3rd-4th degree of radiation pathomorphism in 89.5 +/- 4.5% of the tumors); in the control group in 70.4 +/- 4.4%. Resectability did not differ significantly in both groups. The use of dynamic dose fractionation brought about improved results of combined treatment in general: an increase in the 2-year survival rates by 1.5 himes (59.2 +/- 6.7% as compared to 34.7 +/- 4.2%) was noted.


The Association of GSDMB and ORMDL3 Gene Polymorphisms With Asthma: A Meta-Analysis.
ORM1-like 3 (ORMDL3) belongs to a highly conserved protein family which is anchored as transmembrane protein in the endoplasmic reticulum. Gasdermin B (GSDMB) is adjacent to ORMDL3 on chromosome 17q21.2 and belongs to the gasdermin-domain containing the protein family (GSDM family). Recent reports suggest that GSDMB and ORMDL3 are associated with asthma in several populations. However, genetic association studies that examined the association of GSDMB and ORMDL3 gene variants with asthma showed conflicting results. To assess whether combined evidence shows the association between GSDMB/ORMDL3 polymorphism and asthma. A bibliographic search from MEDLINE identified 13 original articles using the search keywords 'GSDMB', 'ORMDL3', and 'asthma'. An updated literature-based meta-analysis involving 6,691 subjects with asthma, 9,281 control individuals, and 1,360 families were conducted. Meta-odds ratios (ORs) and 95% confidence intervals (CIs) based on the fixed effects model or the random effects model depended on Cochran's Q-statistic and I(2) values. Data from case-control and TDT studies were analyzed in an allelic model using the Catmap software. We selected and identified 3 SNPs of ORMDL3 associated with asthma (rs8076131: OR=1.10; 95% CI, 1.02-1.20; P=0.012. rs12603332: OR=1.15; 95% CI, 1.05-1.25; P=0.002. rs3744246: OR=1.10; 95% CI, 1.02-1.17; P=0.008) and 1 SNP of GSDMB associated with asthma (rs7216389: OR=1.37; 95% CI, 1.27-1.47; P<0.01). Publication bias was estimated using modified Egger's linear regression test proposed by Harbordetal and revealed no evidence of biases. Furthermore, cumulative meta-analysis in chronological order showed the inclination toward significant association for rs7216389 and rs12603332 with continually adding studies, and the inclination toward null-significant association for rs3744246 and rs8076131. Moderate evidence exists for associations of the ORMDL3 rs8076131, rs12603332, and rs3744246 and GSDMB rs7216389 variants with asthma. Large sample size and representative population-based studies and TDT studies with homogeneous asthmatic patients and well-matched controls are warranted to confirm this finding.


Comparative analyses of three-dimensional models of bacterial reaction centers.
Despite the fact that the three-dimensional structure of an integral membrane protein was first determined 20 years ago, structures have been solved for very few membrane proteins. The reaction center is an exception with many mutant and modified structures available from 3 different bacterial species. In order to relate these structures to the function of the reaction center, an accurate assessment of the reliability of the structural models is required. Here we describe the quality of the structures of the bacterial reaction center based upon different criteria, such as evaluation of the geometry of the models and comparison of different models. Overall, the structures are found to be most accurate in the membrane-embedded regions with the periplasmic and cytoplasmic exposed regions having more disorder and differences among the structural models. In general, the cofactors and the surrounding protein regions are among the most accurately determined regions of the protein, except for the secondary quinone and its binding pocket that shows a large variation among structures. The limited accuracy of the secondary quinone is due to its partial occupancy as a consequence of its functional role and to the presence of surface features, including lipids and detergent molecules.