Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease.
Intestinal mesenchymal cells play essential roles in epithelial homeostasis, matrix remodeling, immunity, and inflammation. But the extent of heterogeneity within the colonic mesenchyme in these processes remains unknown. Using unbiased single-cell profiling of over 16,500 colonic mesenchymal cells, we reveal four subsets of fibroblasts expressing divergent transcriptional regulators and functional pathways, in addition to pericytes and myofibroblasts. We identified a niche population located in proximity to epithelial crypts expressing SOX6, F3 (CD142), and WNT genes essential for colonic epithelial stem cell function. In colitis, we observed dysregulation of this niche and emergence of an activated mesenchymal population. This subset expressed TNF superfamily member 14 (TNFSF14), fibroblastic reticular cell-associated genes, IL-33, and Lysyl oxidases. Further, it induced factors that impaired epithelial proliferation and maturation and contributed to oxidative stress and disease severity in vivo. Our work defines how the colonic mesenchyme remodels to fuel inflammation and barrier dysfunction in IBD.


Determinants of Patient Satisfaction in Internal Medicine Resident Continuity Clinics: Findings of the Educational Innovations Project Ambulatory Collaborative.
Many internal medicine programs have reorganized their resident continuity clinics to improve the ambulatory care experience for residents. The effect of this redesign on patient satisfaction is largely unknown. Our multi-institutional, cross-sectional study included 569 internal medicine residents from 11 programs participating in the Educational Innovations Project Ambulatory Collaborative. An 11-item patient satisfaction survey from the Consumer Assessment of Healthcare Providers and Systems was used to assess patient satisfaction, comparing patient satisfaction in traditional models of weekly continuity clinic with 2 new clinic models. We then examined the relationship between patient satisfaction and other practice variables. Patient satisfaction responses related to resident listening and communication skills, knowledge of medical history, perception of adequate visit time, overall rating, and willingness to refer to family and friends were significantly better in the traditional and block continuity models than the combination model. Higher ambulatory workload was associated with reduced patient perception of respect shown by the physician. The percentage of diabetic patients with glycated hemoglobin < 8% was positively correlated with number of visits, knowledge of medical history, perception of respect, and higher scores for recommending the physician to others. The percentage of diabetic patients with low density lipoprotein < 100 mg/dL was positively correlated with the physician showing respect. Patient satisfaction was similar in programs using block design and traditional models for continuity clinic, and both outperformed the combination model programs. There was a delicate balance between workload and patient perception of the physician showing respect. Care outcome measures for diabetic patients were associated with aspects of patient satisfaction.


Prolonged rod dark adaptation in retinitis pigmentosa.
The time course of rod dark adaptation was measured in 12 patients and carriers of retinitis pigmentosa (RP). In contrast to previous studies the rod absolute threshold was determined prior to any exposure to the bleaching light. For seven of the patients and carriers the recovery of rod sensitivity to the prebleach level was prolonged, with the delay ranging from mild to severe. The prolongation appeared to be limited to the late phase of bleaching recovery; the early portion of rod dark adaptation, measured following a weak bleach, was normal. These results suggest a selective impairment in the processes underlying rod dark adaptation in some patients with RP.


Potato agriculture, late blight science, and the molecularization of plant pathology.
By the mid-1980s nucleic-acid based methods were penetrating the farthest reaches of biological science, triggering rivalries among practitioners, altering relationships among subfields, and transforming the research front. This article delivers a "bottom up" analysis of that transformation at work in one important area of biological science, plant pathology, by tracing the "molecularization" of efforts to understand and control one notorious plant disease -- the late blight of potatoes. It mobilizes the research literature of late blight science as a tool through which to trace the changing typography of the research front from 1983 to 2003. During these years molecularization intensified the traditional fragmentation of the late blight research community, even as it dramatically integrated study of the causal organism into broader areas of biology. In these decades the pathogen responsible for late blight, the oomycete "Phytophthora infestans," was discovered to be undergoing massive, frightening, and still largely unexplained genetic diversification -- a circumstance that lends the episode examined here an urgency that reinforces its historiographical significance as a case-study in the molecularization of the biological sciences.


Twelve ways to confirm targets of activity-based probes in plants.
Activity-based probes are powerful tools to interrogate the functional proteome. Their covalent and often irreversible labeling of proteins facilitates the purification, identification and quantification of labeled proteins. However, the detection of labeled proteins often requires a confirmation, especially when unexpected proteins are identified, or to unravel fluorescent activity profiles. Here, we review twelve approaches towards target confirmation, grouped in approaches by direct target detection, target expression or target depletion. We discuss their proper use and limitations and illustrate these approaches with examples from plant science.


Toxoplasmic encephalitis relapse rates with pyrimethamine-based therapy: systematic review and meta-analysis.
Toxoplasmic encephalitis (TE) is caused by Toxoplasma gondii infection and can be a life-threatening disease in immunocompromised patients. This study evaluated the rate of relapse associated with pyrimethamine-based maintenance therapy (i.e. secondary prophylaxis) in patients with human immunodeficiency virus (HIV) or AIDs treated prior to and after the common use (i.e. 1996) of highly active antiretroviral therapy (HAART) (pre-HAART and post-HAART, respectively). PubMed, Google Scholar, and Cochrane databases were searched to 6 June 2016 using search terms: pyrimethamine, Daraprim, Fansidar, Metakelfin, Fansimef, 5-(4-chlorophenyl)-6-ethyl-2,4-pyrimidinediamine, encephalitis, cerebral, toxoplasmosis, toxoplasmic, and gondii. Single-arm cohort, retrospective, and randomized studies were included. Twenty-six studies with 1,596 patients were included in the analysis; twenty pre-HAART (n = 1,228) studies and six post-HAART (n = 368) were performed. Pooled proportions test for pyrimethamine-based therapy from pre-HAART studies indicated a relapse rate of 19.2% and 18.9% from the fixed-effects and random-effects models, respectively. The relapse rate in the post-HAART studies was 11.1% (fixed and random effects). Continuous therapy was suggestive of lower incidence of relapse compared with intermittent therapy in the pre-HAART era (range, 18.7 to 17.3% vs. 20.9 to 25.6%, respectively). These findings indicate that the likelihood of relapse associated with pyrimethamine-based therepy in patients with HIV and TE decreased after the introduction of HAART to approximately 11%. The findings have important implications as relapse may affect a patient's disease severity and prognosis, increase utilization of health care resources, and result in additional health care expenditure.


Urinary-cell mRNA profile and acute cellular rejection in kidney allografts.
The standard test for the diagnosis of acute rejection in kidney transplants is the renal biopsy. Noninvasive tests would be preferable. We prospectively collected 4300 urine specimens from 485 kidney-graft recipients from day 3 through month 12 after transplantation. Messenger RNA (mRNA) levels were measured in urinary cells and correlated with allograft-rejection status with the use of logistic regression. A three-gene signature of 18S ribosomal (rRNA)-normalized measures of CD3ε mRNA and interferon-inducible protein 10 (IP-10) mRNA, and 18S rRNA discriminated between biopsy specimens showing acute cellular rejection and those not showing rejection (area under the curve [AUC], 0.85; 95% confidence interval [CI], 0.78 to 0.91; P<0.001 by receiver-operating-characteristic curve analysis). The cross-validation estimate of the AUC was 0.83 by bootstrap resampling, and the Hosmer-Lemeshow test indicated good fit (P=0.77). In an external-validation data set, the AUC was 0.74 (95% CI, 0.61 to 0.86; P<0.001) and did not differ significantly from the AUC in our primary data set (P=0.13). The signature distinguished acute cellular rejection from acute antibody-mediated rejection and borderline rejection (AUC, 0.78; 95% CI, 0.68 to 0.89; P<0.001). It also distinguished patients who received anti-interleukin-2 receptor antibodies from those who received T-cell-depleting antibodies (P<0.001) and was diagnostic of acute cellular rejection in both groups. Urinary tract infection did not affect the signature (P=0.69). The average trajectory of the signature in repeated urine samples remained below the diagnostic threshold for acute cellular rejection in the group of patients with no rejection, but in the group with rejection, there was a sharp rise during the weeks before the biopsy showing rejection (P<0.001). A molecular signature of CD3ε mRNA, IP-10 mRNA, and 18S rRNA levels in urinary cells appears to be diagnostic and prognostic of acute cellular rejection in kidney allografts. (Funded by the National Institutes of Health and others.).


Organization of neuronal microtubules in the nematode Caenorhabditis elegans.
We have studied the organization of microtubules in neurons of the nematode Caenorhabditis elegans. Six neurons, which we call the microtubule cells, contain bundles of darkly staining microtubules which can be followed easily in serial-section electron micrographs. Reconstruction of individual microtubules in these cells indicate that most, if not all, microtubules are short compared with the length of the cell process. Average microtubule length varies characteristically with cell type. The arrangement of microtubules gives an overall polarity to each bundle: the distal ends of the microtubles are on the outside of the bundle, whereas the proximal ends are preferentially inside. The distal and proximal ends each have a characteristic appearance indicating that these microtubules may have a polarity of their own. Short microtubules in processes of other neurons in C. elegans have also been observed.


CD36 is important for fatty acid and cholesterol uptake by the proximal but not distal intestine.
CD36, a membrane protein that facilitates fatty acid uptake, is highly expressed in the intestine on the luminal surface of enterocytes. Cd36 null (Cd36(-/-)) mice exhibit impaired chylomicron secretion but no overall lipid absorption defect. Because chylomicron production is most efficient proximally we examined whether CD36 function is important for proximal lipid absorption. CD36 levels followed a steep decreasing gradient along three equal-length, proximal to distal intestinal segments (S1-S3). Enterocytes isolated from the small intestines of Cd36(-/-) mice, when compared with wild type counterparts, exhibited reduced uptake of fatty acid (50%) and cholesterol (60%) in S1. The high affinity fatty acid uptake component was missing in Cd36(-/-) cells. Fatty acid incorporation into triglyceride and triglyceride secretion were also reduced in Cd36(-/-) S1 enterocytes. In vivo, proximal absorption was monitored using mass spectrometry from oleic acid enrichment of S1 lipids, 90 min (active absorption) and 5 h (steady state) after intragastric olive oil (70% triolein). Oleate enrichment was 50% reduced at 90 min in Cd36(-/-) tissue consistent with defective uptake whereas no differences were measured at 5 h. In Cd36(-/-) S1, mRNA for L-fabp, Dgat1, and apoA-IV was reduced. Protein levels for FATP4, SR-BI, and NPC1L1 were similar, whereas those for apoB48 and apoA-IV were significantly lower. A large increase in NPC1L1 was observed in Cd36(-/-) S2 and S3. The findings support the role of CD36 in proximal absorption of dietary fatty acid and cholesterol for optimal chylomicron formation, whereas CD36-independent mechanisms predominate in distal segments.


Pyruvate protects pathogenic spirochetes from H2O2 killing.
Pathogenic spirochetes cause clinically relevant diseases in humans and animals, such as Lyme disease and leptospirosis. The causative agent of Lyme disease, Borrelia burgdorferi, and the causative agent of leptospirosis, Leptospria interrogans, encounter reactive oxygen species (ROS) during their enzootic cycles. This report demonstrated that physiologically relevant concentrations of pyruvate, a potent H2O2 scavenger, and provided passive protection to B. burgdorferi and L. interrogans against H2O2. When extracellular pyruvate was absent, both spirochetes were sensitive to a low dose of H2O2 (≈0.6 µM per h) generated by glucose oxidase (GOX). Despite encoding a functional catalase, L. interrogans was more sensitive than B. burgdorferi to H2O2 generated by GOX, which may be due to the inherent resistance of B. burgdorferi because of the virtual absence of intracellular iron. In B. burgdorferi, the nucleotide excision repair (NER) and the DNA mismatch repair (MMR) pathways were important for survival during H2O2 challenge since deletion of the uvrB or the mutS genes enhanced its sensitivity to H2O2 killing; however, the presence of pyruvate fully protected ΔuvrB and ΔmutS from H2O2 killing further demonstrating the importance of pyruvate in protection. These findings demonstrated that pyruvate, in addition to its classical role in central carbon metabolism, serves as an important H2O2 scavenger for pathogenic spirochetes. Furthermore, pyruvate reduced ROS generated by human neutrophils in response to the Toll-like receptor 2 (TLR2) agonist zymosan. In addition, pyruvate reduced neutrophil-derived ROS in response to B. burgdorferi, which also activates host expression through TLR2 signaling. Thus, pathogenic spirochetes may exploit the metabolite pyruvate, present in blood and tissues, to survive H2O2 generated by the host antibacterial response generated during infection.


Striatal Associative Learning Signals Are Tuned to In-groups.
An important feature of adaptive social behavior is the ability to flexibly modify future actions based on the successes or failures of past experiences. The ventral striatum (VS) occupies a central role in shaping behavior by using feedback to evaluate actions and guide learning. The current studies tested whether feedback indicating the need to update social knowledge would engage the VS, thereby facilitating subsequent learning. We also examined the sensitivity of these striatal signals to the value associated with social group membership. Across two fMRI studies, participants answered questions testing their knowledge about the preferences of personally relevant social groups who were high (in-group) or low (out-group) in social value. Participants received feedback indicating whether their responses were correct or incorrect on a trial-by-trial basis. After scanning, participants were given a surprise memory test examining memory for the different types of feedback. VS activity in response to social feedback correlated with subsequent memory, specifying a role for the VS in encoding and updating social knowledge. This effect was more robust in response to in-group than out-group feedback, indicating that the VS tracks variations in social value. These results provide novel evidence of a neurobiological mechanism adaptively tuned to the motivational relevance of the surrounding social environment that focuses learning efforts on the most valuable social outcomes and triggers adjustments in behavior when necessary.


Lymphatic vessels develop during tubulointerstitial fibrosis.
Recent progress with specific markers of lymphatic vessel endothelium allowed recognition of lymphangiogenic events in various disease states; however, there is little information concerning this process in human chronic renal diseases. To determine this we measured expression of the lymphatic marker D2-40 and vascular endothelial growth factor-C (VEGF-C), a major growth factor in lymphangiogenesis, in 124 human renal biopsy specimens. In the kidneys of control subjects and in uninjured areas of pathologic specimens, lymphatic vessels were detected only around the arcuate and interlobular arteries. An increase in the number of lymphatic vessels was found at the site of tubulointerstitial lesions correlating with the degree of tissue damage and more strongly correlating with areas of fibrosis than inflammation. On serial sections, lymphatic vessel proliferation was found in the tubulointerstitial area at the site of tuft adhesions to Bowman's capsule. Lymphatic growth was associated with VEGF-C expression in inflammatory mononuclear cells and tubular epithelial cells, mainly of proximal tubules. Lymphangiogenesis and VEGF-C expression was elevated in diabetic nephropathy in comparison to other renal diseases. Our results indicate that lymphangiogenesis is a common feature in the progression of the tubulointerstitial fibrosis.


Choroidal thickness predicts progression of myopic maculopathy in high myopes: a 2-year longitudinal study.
To prospectively determine the impact of choroidal thickness (CT) on the myopic maculopathy progression. This is a prospective, longitudinal, observational study. In total, 434 participants aged 7-70 years with bilateral high myopia (≤-6 D spherical error, range, -6 to -27.0 D) completed follow-up visits for 2 years. The baseline CT centred on the fovea was measured using a swept-source optical coherence tomography (OCT). Myopic maculopathy progression was determined by fundus photography. Logistic model was used to examine the impact of CT at baseline on the myopic maculopathy progression. Likelihood ratio test was adopted for model comparison. The mean baseline age, spherical equivalence and subfoveal CT (SFCT) of the participants were 23.2±12.5 years, -10.50±3.18 D and 153.20±72.76 μm, respectively. Over 2-year's follow-up, 74 of 434 eyes (17.1%) had myopic maculopathy progression. Baseline SFCT was thinner in eyes with myopic maculopathy progression than those without (67.26±37.67 μm vs 170.95±65.45 μm; mean difference, 99.31 μm; 95% CI 83.61 to 115.01 μm; p<0.001). The same patterns of differences were observed in 7-18 years, 19-39 years and 40-70 years. In multivariate logistic regression model, SFCT was a significant risk factor (adjusted OR=0.97, p<0.005) when age, gender, axial length and baseline myopic maculopathy category were adjusted for. The addition of SFCT significantly improved the predictive discrimination of myopic maculopathy progression in comparison with that included established risk factors alone (area under the receiver operating characteristic curve, 0.899 vs 0.942, p<0.001). CT is an independent predictor for myopic maculopathy progression.


The role of masculinity and distinctiveness in judgments of human male facial attractiveness.
Masculinity and distinctiveness have been found to influence the attractiveness of human male faces. The relationship between masculinity and distinctiveness, however, has received little attention. In Expt 1, we examine how current averaging techniques and manipulated sexual dimorphism influence ratings of attractiveness, masculinity, and distinctiveness. In agreement with previous studies, composite faces were found to be more attractive than individual faces. Averaging resulted in increased ratings of attractiveness but decreased ratings of masculinity and distinctiveness. This supports both that attractiveness is related to averageness and findings showing a preference for feminine traits in male faces. When controlling for attractiveness, no significant relationship was found between masculinity and distinctiveness. Manipulating sexual dimorphism did not alter distinctiveness ratings, indicating that feminized and masculinized faces are equally distinctive. These results are suggestive that masculinity and distinctiveness are separable components in face perception. In Expt 2, we look to improve on previous studies utilizing composite faces by examining how averaging in texture-only or shape-only changes perceptions of attractiveness, masculinity, and distinctiveness. Averaging in both shape and texture were found to increase attractiveness independently, showing that the increased attractiveness of composites is due to the combined action of these two manipulations.


Contributions of phenotypic plasticity to differences in thermogenic performance between highland and lowland deer mice.
Small mammals face especially severe thermoregulatory challenges at high altitude because the reduced O2 availability constrains the capacity for aerobic thermogenesis. Adaptive enhancement of thermogenic performance under hypoxic conditions may be achieved via physiological adjustments that occur within the lifetime of individuals (phenotypic plasticity) and/or genetically based changes that occur across generations, but their relative contributions to performance differences between highland and lowland natives are unclear. Here, we examined potentially evolved differences in thermogenic performance between populations of deer mice (Peromyscus maniculatus) that are native to different altitudes. The purpose of the study was to assess the contribution of phenotypic plasticity to population differences in thermogenic performance under hypoxia. We used a common-garden deacclimation experiment to demonstrate that highland deer mice have enhanced thermogenic capacities under hypoxia, and that performance differences between highland and lowland mice persist when individuals are born and reared under common-garden conditions, suggesting that differences in thermogenic capacity have a genetic basis. Conversely, population differences in thermogenic endurance appear to be entirely attributable to physiological plasticity during adulthood. These combined results reveal distinct sources of phenotypic plasticity for different aspects of thermogenic performance, and suggest that thermogenic capacity and endurance may have different mechanistic underpinnings.


Facile growth of a single-crystal pattern: a case study of HKUST-1.
In order to fabricate metal-organic framework (MOF) based devices, it is desirable to precisely position high-quality and mono-sized MOF crystals on supports. In this work, we demonstrate a facile solution procedure for the fabrication of oriented and monodispersed single-crystal MOF pattern. We expect that such capability will expand the scope of applications of MOFs to advanced fields.


Markov Boundary Discovery with Ridge Regularized Linear Models.
Ridge regularized linear models (RRLMs), such as ridge regression and the SVM, are a popular group of methods that are used in conjunction with coefficient hypothesis testing to discover explanatory variables with a significant multivariate association to a response. However, many investigators are reluctant to draw causal interpretations of the selected variables due to the incomplete knowledge of the capabilities of RRLMs in causal inference. Under reasonable assumptions, we show that a modified form of RRLMs can get "very close" to identifying a subset of the Markov boundary by providing a worst-case bound on the space of possible solutions. The results hold for any convex loss, even when the underlying functional relationship is nonlinear, and the solution is not unique. Our approach combines ideas in Markov boundary and sufficient dimension reduction theory. Experimental results show that the modified RRLMs are competitive against state-of-the-art algorithms in discovering part of the Markov boundary from gene expression data.


A meta-analysis of clinical improvements of general well-being by a standardized Lycium barbarum.
Four randomized, blind, placebo-controlled clinical trials were pooled to study the general effects of oral consumption of Lycium barbarum at 120 mL/day, as a standardized juice, GoChi(®) (FreeLife International, Phoenix, AZ, USA). A questionnaire consisting of symptoms graded 0-5 was given to the participants. For each question, the score changes in the questionnaire between pre- and postintervention were summarized by the standardized mean difference and associated SE to perform the meta-analysis. The change was also characterized into a binary outcome, improved or not, to derive odds ratio (OR) and associated SE derived by a binary outcome using the Mantel-Haenszel method. The meta-analysis and heterogeneity were evaluated with the R program using the rmeta package. Statistical significance was set at 5%. In total, 161 participants (18-72 years old) were included in the meta-analysis. Compared with the placebo group (n=80), the active group (n=81) showed significant improvements in weakness, stress, mental acuity, ease of awakening, shortness of breath, focus on activity, sleep quality, daydreaming, and overall feelings of health and well-being under a random effects model. A fixed effects model showed additional improvements in fatigue, depression, circulation, and calmness. The OR indicated significantly higher chance to improve fatigue, dizziness, and sleep quality. Three studies had statistically significant heterogeneity in procrastination, shoulder stiffness, energy, and calmness. The present meta-analysis confirmed the various health effects of L. barbarum polysaccharides-standardized L. barbarum intake found in the previous randomized, double-blind, placebo-controlled human clinical trials and revealed it resulted in statistically significant improvements in neurological/psychological performance and overall feelings of health and well-being compared with the placebo group under both the fixed and the random effects models of the R program.


Effect of relevant environmental stresses on survival of enterohemorrhagic Escherichia coli in dry-fermented sausage.
Dry-fermented sausages (DFSs) have been linked to several serious foodborne outbreaks of enterohemorrhagic Escherichia coli (EHEC). The ability of pathogens to utilize adaptive responses to different stressful conditions intended to control their growth in foods, food preparation and production processes may enhance their survival. In certain cases, induced tolerance to one type of stress may lead to enhanced resistance to the applied stress as well as to other stresses. We exposed two EHEC strains, MF3582 of serotype O157:H- and MF5554 of serogroup O145, to different stresses commonly encountered during a production process. The two EHEC strains, previously shown to have different abilities to survive DFS production process conditions, were subjected to low temperatures (4°C and 12°C), 5% NaCl or 1% lactic acid for 6days prior to being added to sausage batters. Survival of EHEC was recorded in salami of two recipes, fermented at two temperatures (20°C and 30°C). The results showed that recipe type had the largest impact on EHEC reductions where Moderate recipe (MR) salami batters containing increased levels of NaCl, glucose and NaNO2 provided enhanced EHEC reductions in salami (2.6 log10) compared to Standard recipe (SR) salami (1.7 log10). Effects of pre-exposure stresses were dependent both on strain and recipe. While acid adaptation of MF5554 provided enhanced log10 reductions from 2.0 to 3.0 in MR sausages, adaptation to a combination of acid and salt stress showed the opposite effect in SR sausages with reductions of only 1.1 log10 as compared to the average of 1.8 log10 for the other SR sausages. Otherwise, the salt and acid adaptation single stresses had relatively small effects on EHEC survival through the DFS production process and subsequent storage and freeze/thaw treatments. Growing cells and cells frozen in batter survived poorly in MR sausages with an average reduction of 3.4 and 3.2 log10, respectively. The reductions of EHEC after storage of DFS increased with higher temperature and storage time. Up to 3.7 log10 additional reduction was obtained when MF3582 was stored for 2months at 20°C. In conclusion, adaptation of EHEC to acid, salt and low temperatures prior to being introduced in a DFS production process has limited, but strain dependent effects on EHEC reductions. Producers should avoid conditions leading to acid and salt adapted cells that can contaminate the sausage batter. Recipe parameters had the largest impact on EHEC reductions while storage at 20°C is effective for enhanced reductions in finished products.


Dynamic whole-body PET parametric imaging: II. Task-oriented statistical estimation.
In the context of oncology, dynamic PET imaging coupled with standard graphical linear analysis has been previously employed to enable quantitative estimation of tracer kinetic parameters of physiological interest at the voxel level, thus, enabling quantitative PET parametric imaging. However, dynamic PET acquisition protocols have been confined to the limited axial field-of-view (~15-20 cm) of a single-bed position and have not been translated to the whole-body clinical imaging domain. On the contrary, standardized uptake value (SUV) PET imaging, considered as the routine approach in clinical oncology, commonly involves multi-bed acquisitions, but is performed statically, thus not allowing for dynamic tracking of the tracer distribution. Here, we pursue a transition to dynamic whole-body PET parametric imaging, by presenting, within a unified framework, clinically feasible multi-bed dynamic PET acquisition protocols and parametric imaging methods. In a companion study, we presented a novel clinically feasible dynamic (4D) multi-bed PET acquisition protocol as well as the concept of whole-body PET parametric imaging employing Patlak ordinary least squares (OLS) regression to estimate the quantitative parameters of tracer uptake rate Ki and total blood distribution volume V. In the present study, we propose an advanced hybrid linear regression framework, driven by Patlak kinetic voxel correlations, to achieve superior trade-off between contrast-to-noise ratio (CNR) and mean squared error (MSE) than provided by OLS for the final Ki parametric images, enabling task-based performance optimization. Overall, whether the observer's task is to detect a tumor or quantitatively assess treatment response, the proposed statistical estimation framework can be adapted to satisfy the specific task performance criteria, by adjusting the Patlak correlation-coefficient (WR) reference value. The multi-bed dynamic acquisition protocol, as optimized in the preceding companion study, was employed along with extensive Monte Carlo simulations and an initial clinical (18)F-deoxyglucose patient dataset to validate and demonstrate the potential of the proposed statistical estimation methods. Both simulated and clinical results suggest that hybrid regression in the context of whole-body Patlak Ki imaging considerably reduces MSE without compromising high CNR. Alternatively, for a given CNR, hybrid regression enables larger reductions than OLS in the number of dynamic frames per bed, allowing for even shorter acquisitions of ~30 min, thus further contributing to the clinical adoption of the proposed framework. Compared to the SUV approach, whole-body parametric imaging can provide better tumor quantification, and can act as a complement to SUV, for the task of tumor detection.


The character strengths of class clowns.
Class clowns traditionally were studied as a type concept and identified via sociometric procedures. In the present study a variable-centered approach was favored and class clown behaviors were studied in the context of character strengths, orientations to happiness and satisfaction with life. A sample of 672 Swiss children and adolescents filled in an 18 item self-report instrument depicting class clown behaviors. A hierarchical model of class clown behaviors was developed distinguishing a general factor and the four positively correlated dimensions of "identified as a class clown," "comic talent," "disruptive rule-breaker," and "subversive joker." Analysis of the general factor showed that class clowns were primarily male, and tended to be seen as class clowns by the teacher. Analyses of the 24 character strengths of the VIA-Youth (Park and Peterson, 2006) showed that class clowns were high in humor and leadership, and low in strengths like prudence, self-regulation, modesty, honesty, fairness, perseverance, and love of learning. An inspection of signature strengths revealed that 75% of class clowns had humor as a signature strength. Furthermore, class clown behaviors were generally shown by students indulging in a life of pleasure, but low life of engagement. The four dimensions yielded different character strengths profiles. While all dimensions of class clowns behaviors were low in temperance strengths, the factors "identified as the class clown" and "comic talent" were correlated with leadership strengths and the two negative factors ("disruptive rule-breaker," "subversive joker") were low in other directed strengths. The disruptive rule breaking class clown was additionally low in intellectual strengths. While humor predicted life satisfaction, class clowning tended to go along with diminished satisfaction with life. It is concluded that different types of class clowns need to be kept apart and need different attention by teachers.


Whole-exome sequencing of pancreatic cancer defines genetic diversity and therapeutic targets.
Pancreatic ductal adenocarcinoma (PDA) has a dismal prognosis and insights into both disease etiology and targeted intervention are needed. A total of 109 micro-dissected PDA cases were subjected to whole-exome sequencing. Microdissection enriches tumour cellularity and enhances mutation calling. Here we show that environmental stress and alterations in DNA repair genes associate with distinct mutation spectra. Copy number alterations target multiple tumour suppressive/oncogenic loci; however, amplification of MYC is uniquely associated with poor outcome and adenosquamous subtype. We identify multiple novel mutated genes in PDA, with select genes harbouring prognostic significance. RBM10 mutations associate with longer survival in spite of histological features of aggressive disease. KRAS mutations are observed in >90% of cases, but codon Q61 alleles are selectively associated with improved survival. Oncogenic BRAF mutations are mutually exclusive with KRAS and define sensitivity to vemurafenib in PDA models. High-frequency alterations in Wnt signalling, chromatin remodelling, Hedgehog signalling, DNA repair and cell cycle processes are observed. Together, these data delineate new genetic diversity of PDA and provide insights into prognostic determinants and therapeutic targets.


Congenital myasthenic syndrome due to mutations in MUSK suggests that the level of MuSK phosphorylation is crucial for governing synaptic structure.
MUSK encodes the muscle-specific receptor tyrosine kinase (MuSK), a key component of the agrin-LRP4-MuSK-DOK7 signaling pathway, which is essential for the formation and maintenance of highly specialized synapses between motor neurons and muscle fibers. We report a patient with severe early-onset congenital myasthenic syndrome and two novel missense mutations in MUSK (p.C317R and p.A617V). Functional studies show that MUSK p.C317R, located at the frizzled-like cysteine-rich domain of MuSK, disrupts an integral part of MuSK architecture resulting in ablated MuSK phosphorylation and acetylcholine receptor (AChR) cluster formation. MUSK p.A617V, located at the kinase domain of MuSK, enhances MuSK phosphorylation resulting in anomalous AChR cluster formation. The identification and evidence for pathogenicity of MUSK mutations supported the initiation of treatment with β2-adrenergic agonists with a dramatic improvement of muscle strength in the patient. This work suggests uncharacterized mechanisms in which control of the precise level of MuSK phosphorylation is crucial in governing synaptic structure.


Hedgehog signaling blockade delays hepatocarcinogenesis induced by hepatitis B virus X protein.
The hepatitis B virus (HBV) encoded X protein (HBx) contributes centrally to the pathogenesis of hepatocellular carcinoma (HCC). Aberrant activation of the Hedgehog (Hh) pathway has been linked to many tumor types including HCC. Thus, experiments were designed to test the hypothesis that HBx promotes HCC via activation of Hh signaling. HBx expression correlated with an upregulation of Hh markers in human liver cancer cell lines, in liver samples from HBV infected patients with HCC, and in the livers of HBx transgenic mice (HBxTg) that develop hepatitis, steatosis, and dysplasia, culminating in the appearance of HCC. The findings in human samples provide clinical validation for the in vitro results and those in the HBxTg. Blockade of Hh signaling inhibited HBx stimulation of cell migration, anchorage-independent growth, tumor development in HBxTg, and xenograft growth in nude mice. Results suggest that the ability of HBx to promote cancer is at least partially dependent upon the activation of the Hh pathway. This study provides biologic evidence for the role of Hh signaling in the pathogenesis of HBV-mediated HCC and suggests cause and effect for the first time. The observation that inhibition of Hh signaling partially blocked the ability of HBx to promote growth and migration in vitro and tumorigenesis in two animal models implies that Hh signaling may represent an "oncogene addiction" pathway for HBV-associated HCC. This work could be central to designing specific treatments that target early development and progression of HBx-mediated HCC.


Peri-operative mortality in the anaesthetic service at Tygerberg Hospital.
This study reports on the process of peer review of peri-operative mortality at Tygerberg Hospital. The peri-operative mortality rate for the past 3.5 years was 11.9/10,000. The departmental evaluation committee thought that 3.4/10,000 of these deaths were anaesthetic-related. In 2.3/10,000, the anaesthetic contributed to the death of the patient and in 10 cases (total case load = 94,945; i.e. 1.1/10,000) anaesthesia was responsible for the death of the patient. The majority of the peri-operative deaths (8.5/10,000) were caused by a combination of trauma and haemorrhagic shock.


AQP5 Variants Affect Tumoral Expression of AQP5 and Survival in Patients with Early Breast Cancer.
Our previous study showed the association of AQP5 upregulation with cancer proliferation and migration in breast cancer cell lines and with unfavorable prognosis in patients with early breast cancer (EBC). In the current study, we analyzed the association of AQP5 variants or their haplotypes with AQP5 expression and their prognostic impact for survival in patients with EBC. Three AQP5 polymorphisms (rs74091166, rs3736309, and rs1964676) were selected based on the SNP database and genotyped using the Sequenom MassARRAY in 374 out of 447 patients with EBC in whom AQP5 expression had been investigated in our previous study. The allele frequencies of the selected variants in the current study were similar to those from Asian data previously reported. In a univariate analysis, both rs74091166 and rs1964676 were statistically associated with survival as a dominant model of minor allele. Moreover, a multivariate survival analysis revealed that the CC genotype of rs1964676 is an independent prognostic marker of survival in EBC patients, regardless of stage, tumor subtype, and adjuvant treatment [hazard ratio = 0.399, 0.384, and 0.205; p = 0.021, 0.027, and 0.016 for disease-free survival (DFS), distant DFS, and disease-specific survival, respectively]. In particular, the CT/TT genotype of rs1964676 showed an association with strong expression of AQP5 (58.6 vs. 26.0%; p = 0.001), without any associations with clinical or pathological characteristics including tumor subtype, stage, or histologic grade. The current study suggests AQP5 rs1964676 as a new potential prognostic marker in patients with EBC involved in AQP5 expression.


Secreted Gaussia luciferase as a sensitive reporter gene for in vivo and ex vivo studies of airway gene transfer.
The cationic lipid GL67A is one of the more efficient non-viral gene transfer agents (GTAs) for the lungs, and is currently being evaluated in an extensive clinical trial programme for cystic fibrosis gene therapy. Despite conferring significant expression of vector-specific mRNA following transfection of differentiated human airway cells cultured on air liquid interfaces (ALI) cultures and nebulisation into sheep lung in vivo we were unable to detect robust levels of the standard reporter gene Firefly luciferase (FLuc). Recently a novel secreted luciferase isolated from Gaussia princeps (GLuc) has been described. Here, we show that (1) GLuc is a more sensitive reporter gene and offers significant advantages over the traditionally used FLuc in pre-clinical models for lung gene transfer that are difficult to transfect, (2) GL67A-mediated gene transfection leads to significant production of recombinant protein in these models, (3) promoter activity in ALI cultures mimics published in vivo data and these cultures may, therefore, be suitable to characterise promoter activity in a human ex vivo airway model and (4) detection of GLuc in large animal broncho-alveolar lavage fluid and serum facilitates assessment of duration of gene expression after gene transfer to the lungs. In summary, we have shown here that GLuc is a sensitive reporter gene and is particularly useful for monitoring gene transfer in difficult to transfect models of the airway and lung. This has allowed us to validate that GL67A, which is currently in clinical use, can generate significant amounts of recombinant protein in fully differentiated human air liquid interface cultures and the ovine lung in vivo.


Disparities in outcomes for African American and Latino subjects in the Flexible Initial Retrovirus Suppressive Therapies (FIRST) trial.
To benefit maximally from antiretroviral therapy, patients with HIV infection must enter care before their disease is advanced and adhere to care. We sought to determine if and where on this continuum of care racial/ethnic disparities were evident. Data from the Flexible Initial Retrovirus Suppressive Therapies (FIRST) trial, which evaluated three strategies for initial HIV therapy, were compared for White, African American, and Latino subjects. Outcomes included progression of disease and death, HIV viral suppression, and change in CD4(+) cell count. Multivariate Cox proportional hazard models adjusted for known predictors of survival. There were 1357 subjects, including 368 non-Latino white, 751 non-Latino African American, and 238 Latino subjects. At baseline, the two latter groups were more likely to have had AIDS and had lower CD4(+) cell counts than white subjects. In follow-up, African American subjects had lower self-reported adherence to therapy, lower CD4(+) cell count increases, and lower odds of viral suppression. African American and Latino subjects had unadjusted hazard ratios of progression of disease or death of 1.57 (1.17, 2.10; p = 0.0025) and 1.57 (1.09, 2.26; p = 0.02), respectively. Adjusting for baseline differences and differences in adherence, CD4(+) cell count change, and viral suppression accounted for the disparities in outcomes. Opportunities to reduce disparities in outcomes for African American and Latino patients exist along the continuum of HIV care. Efforts to promote access to HIV testing and care and to improve adherence have the potential to reduce racial/ethnic disparities in outcomes of patients with HIV infection.


Misexpression of a chloroplast aspartyl protease leads to severe growth defects and alters carbohydrate metabolism in Arabidopsis.
The crucial role of carbohydrate in plant growth and morphogenesis is widely recognized. In this study, we describe the characterization of nana, a dwarf Arabidopsis (Arabidopsis thaliana) mutant impaired in carbohydrate metabolism. We show that the nana dwarf phenotype was accompanied by altered leaf morphology and a delayed flowering time. Our genetic and molecular data indicate that the mutation in nana is due to a transfer DNA insertion in the promoter region of a gene encoding a chloroplast-located aspartyl protease that alters its pattern of expression. Overexpression of the gene (oxNANA) phenocopies the mutation. Both nana and oxNANA display alterations in carbohydrate content, and the extent of these changes varies depending on growth light intensity. In particular, in low light, soluble sugar levels are lower and do not show the daily fluctuations observed in wild-type plants. Moreover, nana and oxNANA are defective in the expression of some genes implicated in sugar metabolism and photosynthetic light harvesting. Interestingly, some chloroplast-encoded genes as well as genes whose products seem to be involved in retrograde signaling appear to be down-regulated. These findings suggest that the NANA aspartic protease has an important regulatory function in chloroplasts that not only influences photosynthetic carbon metabolism but also plastid and nuclear gene expression.


Inherited determinants of ovarian cancer survival.
Due to variation of outcome among cases, we sought to examine whether overall survival in ovarian cancer was associated with common inherited variants in 227 candidate genes from ovarian cancer-related pathways including angiogenesis, inflammation, detoxification, glycosylation, one-carbon transfer, apoptosis, cell cycle regulation, and cellular senescence. Blood samples were obtained from 325 women with invasive epithelial ovarian cancer diagnosed at the Mayo Clinic from 1999 to 2006. During a median follow-up of 3.8 years (range, 0.1-8.6 years), 157 deaths were observed. Germline DNA was analyzed at 1,416 single nucleotide polymorphisms (SNP). For all patients, and for 203 with serous subtype, we assessed the overall significance of each gene and pathway, and estimated risk of death via hazard ratios (HR) and 95% confidence intervals (CI), adjusting for known prognostic factors. Variation within angiogenesis was most strongly associated with survival time overall (P = 0.03) and among patients with serous cancer (P = 0.05), particularly for EIF2B5 rs4912474 (all patients HR, 0.69; 95% CI, 0.54-0.89; P = 0.004), VEGFC rs17697305 (serous subtype HR, 2.29; 95% CI, 1.34-3.92; P = 0.003), and four SNPs in VHL. Variation within the inflammation pathway was borderline significant (all patients, P = 0.09), and SNPs in CCR3, IL1B, IL18, CCL2, and ALOX5 which correlated with survival time are worthy of follow-up. An extensive multiple-pathway assessment found evidence that inherited differences may play a role in outcome of ovarian cancer patients, particularly in genes within the angiogenesis and inflammation pathways. Our work supports efforts to target such mediators for therapeutic gain.


Cyclosporine A in juvenile idiopathic arthritis. Results of the PRCSG/PRINTO phase IV post marketing surveillance study.
To investigate the clinical use patterns, clinical effect and safety of cyclosporine A (CSA) in juvenile idiopathic arthritis (JIA) in the setting of routine clinical care. An open-ended, phase IV post marketing surveillance study was conducted among members of the Pediatric Rheumatology Collaborative Study Group (PRCSG) and of the Paediatric Rheumatology International Trials Organisation (PRINTO) to identify patients with polyarticular course JIA who had received CSA during the course of their disease. A total of 329 patients, half of whom had systemic JIA, were collected in 21 countries. Data were collected during 1240 routine clinic visits. CSA was started at a mean of 5.8 years after disease onset and was given at a mean dose of 3.4 mg/kg/day. The drug was administered in combination with MTX in 61% and along with prednisone in 65% of the patients who were still receiving CSA. Among patients who were still receiving CSA therapy at the last reported visit, remission was documented in 9% of the patients, whereas in 61% of the patients the disease activity was rated as moderate or severe. The most frequent reason for discontinuation of CSA was insufficient therapeutic effect (61% of the patients); only 10% of the patients stopped CSA because of remission. In 17% of the patients, side effects of therapy was given as the primary reason for discontinuation. This survey suggests that CSA may have a less favourable efficacy profile than MTX and etanercept, whereas the frequency of side effects may be similar. The exact place of CSA in the treatment of JIA can only be established via controlled clinical trial.


Determination of monolayer-protected gold nanoparticle ligand-shell morphology using NMR.
It is accepted that the ligand shell morphology of nanoparticles coated with a monolayer of molecules can be partly responsible for important properties such as cell membrane penetration and wetting. When binary mixtures of molecules coat a nanoparticle, they can arrange randomly or separate into domains, for example, forming Janus, patchy or striped particles. To date, there is no straightforward method for the determination of such structures. Here we show that a combination of one-dimensional and two-dimensional NMR can be used to determine the ligand shell structure of a series of particles covered with aliphatic and aromatic ligands of varying composition. This approach is a powerful way to determine the ligand shell structure of patchy particles; it has the limitation of needing a whole series of compositions and ligands' combinations with NMR peaks well separated and whose shifts due to the surrounding environment can be large enough.


Evaluation of the vertebrobasilar system in thoracic aortic surgery.
We evaluated the probability of vertebrobasilar system malperfusion due to occlusion of the left subclavian artery as assessed by preoperative magnetic resonance angiography in patients scheduled to undergo thoracic aortic surgery. (Study 1) From January 2000 through March 2009, we studied variations of vertebral arteries in 301 patients scheduled to undergo thoracic aortic surgery. We classified vertebral artery variations into 3 categories according to the findings on preoperative magnetic resonance angiography: connection type, interrupted right vertebral artery, and interrupted left vertebral artery. (Study 2) From February 2007 through January 2010, we evaluated the cerebral complication in 41 patients who had occlusion of the left subclavian artery with a stent graft. (Study 1) On preoperative magnetic resonance angiography, the vertebral artery was classified as connection type in 247 patients, interrupted right vertebral artery in 34, and interrupted left vertebral artery in 20. (Study 2) We performed subclavian obstruction test, left-right subclavian artery bypass, or left subclavian artery-left common carotid artery bypass to the 3 patients with interrupted right vertebral artery, respectively. Forty patients (98%) out of 41 patients had no complication after occlusion of the left subclavian artery. Preoperative magnetic resonance angiography is useful for detection of the patients with high risk of vertebrobasilar system malperfusion due to occlusion of the left subclavian artery.


Pertussis toxin inhibits neutrophil recruitment to delay antibody-mediated clearance of Bordetella pertussis.
Whooping cough is considered a childhood disease, although there is growing evidence that children are infected by adult carriers. Additionally, increasing numbers of vaccinated adults are being diagnosed with Bordetella pertussis disease. Thus it is critical to understand how B. pertussis remains endemic even in highly vaccinated or immune populations. Here we used the mouse model to examine the nature of sterilizing immunity to B. pertussis. Antibodies were necessary to control infection but did not rapidly clear B. pertussis from the lungs. However, antibodies affected B. pertussis after a delay of at least a week by a mechanism that involved neutrophils and Fc receptors, suggesting that neutrophils phagocytose and clear antibody-opsonized bacteria via Fc receptors. B. pertussis blocked migration of neutrophils and inhibited their recruitment to the lungs during the first week of infection by a pertussis toxin-dependent (PTx-dependent) mechanism; a PTx mutant of B. pertussis induced rapid neutrophil recruitment and was rapidly cleared from the lungs by adoptively transferred antibodies. Depletion of neutrophils abrogated the defects of the PTx mutant. Together these results indicate that PTx inhibits neutrophil recruitment, which consequently allows B. pertussis to avoid rapid antibody-mediated clearance and therefore successfully infect immune hosts.


Human PIF helicase is cell cycle regulated and associates with telomerase.
The evolutionarily conserved PIF1 DNA helicase family is important for the maintenance of genome stability in the yeast, Saccharomyces cerevisiae. There are two PIF1 family helicases in S. cerevisiae, Pif1p and Rrm3p that both possess 5'-->3' DNA helicase activity but maintain unique functions in telomerase regulation and semi-conservative DNA replication. Database analysis shows that the PIF1 helicase family is represented by a single homologue in higher eukaryotes. To analyze the function of PIF1 homologues in mammals, we cloned the full length human PIF (hPIF) cDNA. Comparison of hPIF with its S. cerevisiae homologues showed that human PIF is equally similar to Pif1p and Rrm3p. Human PIF was expressed at low levels in a variety of tissues and immunofluorescence analysis showed that ectopic hPIF was localized to nuclear foci. hPIF was expressed in late S/G2 phase of the cell cycle and this cell cycle regulated abundance was conferred by both cell cycle regulated mRNA accumulation and ubiquitin-mediated degradation. Furthermore, hPIF is likely a target of the anaphase promoting complex/cyclosome as its abundance was decreased when an activator of the APC/C was overexpressed. Finally, antibodies against hPIF immunoprecipitated telomerase activity from human cell lines, and we have observed a physical interaction between hPIF and the catalytic subunit of telomerase, hTERT. Our data suggest that human PIF, like S. cerevisiae Pif1p, plays a role in telomerase regulation.


Climate Change Impact: The Experience of the Coastal Areas of Bangladesh Affected by Cyclones Sidr and Aila.
Bangladesh is considered one of the countries most at risk to the effects of climate change and its coastal area is most vulnerable. This study tries to explore the experiences of cyclones Sidr and Aila affected people living in the coastal areas of Bangladesh. This study was conducted in the cyclone Sidr affected Amtali Upazila of Barguna District and in the cyclone Aila affected Koyra Upazila of Khulna District. Primary data collection was done using Focus Group Interview and then a thematic analysis approach was used for analysis. Three core themes emerged from the analysis and they are, firstly, impacts of climate change on the socioeconomic condition of the people, secondly, the impact on the health status of the population, and finally the impact on vulnerable people. Findings show that the effects of climate change have serious consequences on the livelihood patterns of the affected population and on their overall health status. As a result, the unfavorable health condition of these affected people makes them more vulnerable to various emerging diseases.


Genome-wide association study of brain amyloid deposition as measured by Pittsburgh Compound-B (PiB)-PET imaging.
Deposition of amyloid plaques in the brain is one of the two main pathological hallmarks of Alzheimer's disease (AD). Amyloid positron emission tomography (PET) is a neuroimaging tool that selectively detects in vivo amyloid deposition in the brain and is a reliable endophenotype for AD that complements cerebrospinal fluid biomarkers with regional information. We measured in vivo amyloid deposition in the brains of ~1000 subjects from three collaborative AD centers and ADNI using 11C-labeled Pittsburgh Compound-B (PiB)-PET imaging followed by meta-analysis of genome-wide association studies, first to our knowledge for PiB-PET, to identify novel genetic loci for this endophenotype. The APOE region showed the most significant association where several SNPs surpassed the genome-wide significant threshold, with APOE*4 being most significant (P-meta = 9.09E-30; β = 0.18). Interestingly, after conditioning on APOE*4, 14 SNPs remained significant at P < 0.05 in the APOE region that were not in linkage disequilibrium with APOE*4. Outside the APOE region, the meta-analysis revealed 15 non-APOE loci with P < 1E-05 on nine chromosomes, with two most significant SNPs on chromosomes 8 (P-meta = 4.87E-07) and 3 (P-meta = 9.69E-07). Functional analyses of these SNPs indicate their potential relevance with AD pathogenesis. Top 15 non-APOE SNPs along with APOE*4 explained 25-35% of the amyloid variance in different datasets, of which 14-17% was explained by APOE*4 alone. In conclusion, we have identified novel signals in APOE and non-APOE regions that affect amyloid deposition in the brain. Our data also highlights the presence of yet to be discovered variants that may be responsible for the unexplained genetic variance of amyloid deposition.


Appropriateness of healthcare interventions: concepts and scoping of the published literature.
This report is a scoping review of the literature with the objective of identifying definitions, conceptual models and frameworks, as well as the methods and range of perspectives, for determining appropriateness in the context of healthcare delivery. To lay groundwork for future, intervention-specific research on appropriateness, this work was carried out as a scoping review of published literature since 1966. Two reviewers, with two screens using inclusion/exclusion criteria based on the objective, focused the research and articles chosen for review. The first screen examined 2,829 abstracts/titles, with the second screen examining 124 full articles, leaving 37 articles deemed highly relevant for data extraction and interpretation. Appropriateness is defined largely in terms of net clinical benefit to the average patient and varies by service and setting. The most widely used method to assess appropriateness of healthcare services is the RAND/UCLA Model. There are many related concepts such as medical necessity and small-areas variation. A broader approach to determining appropriateness for healthcare interventions is possible and would involve clinical, patient and societal perspectives.


Fronto-temporal dysregulation in asymptomatic bipolar I patients: a paired associate functional MRI study.
Bipolar disorder is associated with persistent declarative memory disturbances, but the neural basis of these deficits is not well understood. We used fMRI to investigate brain activity during performance on a face-name paired associate task, which allows for the dissociation of encoding and recall-related memory processes. Fifteen clinically remitted bipolar I disorder patients and 24 demographically matched healthy comparison subjects were scanned during task performance. At the voxel level, bipolar patients showed reduced cortical activation, relative to controls, in multiple task-related brain regions during encoding. During recognition, bipolar patients under-activated left hippocampal and parahippocampal regions, despite adequate task performance. Region of interest analyses indicated that, during encoding, bipolar patients had greater bilateral dorsolateral prefrontal (DLPFC) activity than healthy subjects. In contrast, during recognition patients showed hypo-activation relative to controls in the right, but not the left, DLPFC. Although hippocampal activity did not differ between groups during encoding, bipolar patients failed to activate hippocampal regions to the same extent as healthy subjects during recognition. Finally, while better task performance was associated with recognition-related hippocampal activity in healthy subjects, bipolar patients showed an inverse relationship between task performance and hippocampal activity. Remitted bipolar patients over-engaged dorsolateral prefrontal regions when learning face-name pairs, but relative hypoactivation in both prefrontal and medial temporal regions during recognition. These findings suggest a neural basis for the long-term memory deficits consistently observed in patients with bipolar disorder; further, as these patterns appear in symptomatically remitted patients, they are unlikely to be an artifact of mood symptoms.


Vibratory Dynamics of Four Types of Excised Larynx Phonations.
There are four types of signals that are typical representations of vocal fold vibratory patterns. Type 1 signals are nearly periodic, type 2 signals contain subharmonic properties, type 3 signals are chaotic, and type 4 signals are characterized as white noise. High-speed imaging allows detailed observation of these vocal fold vibratory patterns. Therefore, high-speed imaging can explore the vibratory mechanism behind each of the four types of signals. The glottal area time series of the four types of vocal fold vibrations were calculated from high-speed images of 10 excised canine larynges. Nonlinear dynamic parameters of correlation dimension (D2) and Kolmogorov entropy (K2) were used to quantify the characteristics of the glottal areas and acoustical signals for each voice signal type. The correlation dimension and Kolmogorov entropy of the glottal areas and acoustical signals for type 1, 2, and 3 voice signals were consistent with the results of previous studies. Interestingly, there was a difference between the glottal area and acoustical signals of type 4 voice signals (P < 0.001). Both the correlation dimension and Kolmogorov entropy of the type 4 glottal area were close to 0. In contrast, the type 4 acoustical signals had an infinite correlation dimension and a Kolmogorov entropy that was close to 1. Turbulence in the vocal tract creates high-frequency breathiness, causing noise in the acoustical signal of type 4 voice, proving that the acoustical signal does not represent the motion mechanism behind type 4 voice. The results of this study demonstrate that high-speed imaging can provide a more accurate representation of the type 4 vocal fold vibratory pattern, and a more effective method to explore the mechanism of type 4 signals.


Selective determinants of inositol 1,4,5-trisphosphate and adenophostin A interactions with type 1 inositol 1,4,5-trisphosphate receptors.
Adenophostin A (AdA) is a potent agonist of inositol 1,4,5-trisphosphate receptors (IP(3) R). AdA shares with IP(3) the essential features of all IP(3) R agonists, namely structures equivalent to the 4,5-bisphosphate and 6-hydroxyl of IP(3) , but the basis of its increased affinity is unclear. Hitherto, the 2'-phosphate of AdA has been thought to provide a supra-optimal mimic of the 1-phosphate of IP(3) . We examined the structural determinants of AdA binding to type 1 IP(3) R (IP(3) R1). Chemical synthesis and mutational analysis of IP(3) R1 were combined with (3) H-IP(3) binding to full-length IP(3) R1 and its N-terminal fragments, and Ca(2+) release assays from recombinant IP(3) R1 expressed in DT40 cells. Adenophostin A is at least 12-fold more potent than IP(3) in functional assays, and the IP(3) -binding core (IBC, residues 224-604 of IP(3) R1) is sufficient for this high-affinity binding of AdA. Removal of the 2'-phosphate from AdA (to give 2'-dephospho-AdA) had significantly lesser effects on its affinity for the IBC than did removal of the 1-phosphate from IP(3) (to give inositol 4,5-bisphosphate). Mutation of the only residue (R568) that interacts directly with the 1-phosphate of IP(3) decreased similarly (by ~30-fold) the affinity for IP(3) and AdA, but mutating R504, which has been proposed to form a cation-π interaction with the adenine of AdA, more profoundly reduced the affinity of IP(3) R for AdA (353-fold) than for IP(3) (13-fold). The 2'-phosphate of AdA is not a major determinant of its high affinity. R504 in the receptor, most likely via a cation-π interaction, contributes specifically to AdA binding.


Assessing the effect of lexical variables in backward recall.
In a recent study, Bireta et al. (2010) suggested that when participants are required to recall lists of items in the reverse order, more attention is devoted to the recall of order at the expense of item information, leading to the abolition of item-based phenomena (the item and order trade-off hypothesis). In order to test the item and order trade-off hypothesis, we manipulated 4 lexical factors that are well known to influence item retention. The effects of word frequency, of lexicality, of semantic similarity, and of imageability were tested in forward and backward recall. All 4 phenomena were maintained in backward recall, which contradicts the item and order trade-off hypothesis. Instead, we suggest that backward recall might rely on semantic retrieval strategies.


Evidence for disseminated Mycoplasma fermentans in New Jersey residents with antecedent tick attachment and subsequent musculoskeletal symptoms.
Mycoplasma species are one of nature's most abundant groups of microbes. These bacteria inhabit a wide diversity of insect, plant, and animal species, including humans. Certain mycoplasma species have been identified in blood-sucking arthropods, including Ixodes ticks. Frequent human exposure to this genus of ticks led us to explore the possibility of tick-mediated transmission of these bacteria. We evaluated 7 residents of central New Jersey who developed fatigue, musculoskeletal symptoms, and cognitive disturbance after tick attachment. All 7 of these patients lacked both serological evidence and erythema migrans skin lesions characteristic of Lyme disease. We were able to amplify and quantitate Mycoplasma fermentans-specific DNA from their peripheral blood lymphocytes. After antimicrobial therapy, symptoms subsided, and M. fermentans DNA could no longer be detected in their blood specimens. These findings suggest that a subset of disseminated M. fermentans infections may be a vector-mediated process in humans and should be considered in patients with puzzling musculoskeletal presentations.


Randomly amplified polymorphic DNA analysis of Erysipelothrix spp.
The usefulness of randomly amplified polymorphic DNA method (RAPD) to identify each species of genus Erysipelothrix and for epidemiological analysis of this genus was studied. Eighty-one strains and 18 random primers were tested. Among the tested primers, the primers NK51 (GGTGGTGGTATC) and NK6 (CCCGCGCCCC) produced noticeable results. The primer NK51 revealed four species-specific RAPD patterns. Of the 66 strains of E. rhusiopathiae, 64 had the same unique band of 884 bp. Of the 12 strains of E. tonsillarum, 11 produced a 1,265-bp band. In addition, two strains, previously thought to be E. rhusiopathiae, produced the 1,265-bp band, suggesting that they had been misclassified. One strain of E. tonsillarum produced the 884-bp band, suggesting that it too was E. rhusiopathiae. The E. rhusiopathiae strain of serovar 13 produced a 650-bp band, and the strain of serovar 18 produced a clear 420-bp band as well as three weak bands of 1,265, 918, and 444 bp. The primer NK6 revealed 14 RAPD patterns that were not serovar specific. However, different patterns were produced among strains of the same serovar showing that the RAPD method is able to identify the genetic variations of strains of this genus and can rapidly and easily differentiate strains of the same serovar. Based on these results, we concluded that the RAPD method with primers NK51 and NK6 is a rapid and reliable method to identify the species of this genus; we also concluded that this method might be a useful tool for the epidemiological analysis of the Erysipelothrix species.


The role of the hippocampus in solving the Morris water maze.
We suggest that the hippocampus plays two roles that allow rodents to solve the hidden-platform water maze: self-localization and route replay. When an animal explores an environment such as the water maze, the combination of place fields and correlational (Hebbian) long-term potentiation produces a weight matrix in the CA3 recurrent collaterals such that cells with overlapping place fields are more strongly interconnected than cells with nonoverlapping fields. When combined with global inhibition, this forms an attractor with coherent representations of position as stable states. When biased by local view information, this allows the animal to determine its position relative to the goal when it returns to the environment. We call this self-localization. When an animal traces specific routes within an environment, the weights in the CA3 recurrent collaterals become asymmetric. We show that this stores these routes in the recurrent collaterals. When primed with noise in the absence of sensory input, a coherent representation of position still forms in the CA3 population, but then that representation drifts, retracing a route. We show that these two mechanisms can coexist and form a basis for memory consolidation, explaining the anterograde and limited retrograde amnesia seen following hippocampal lesions.


Percutaneous sclerotherapy for venous malformations using polidocanol under fluoroscopy.
This retrospective study evaluated the safety and efficacy of using polidocanol with X-ray fluoroscopy for percutaneous sclerotherapy of venous malformations of the limbs, head, and neck. The subjects were 16 of 18 patients who presented to our department with venous malformations. Two patients were excluded because they were unlikely to benefit from the treatment. Of the 16 included in the study, 1 could not be treated because of inaccessibility, and another was lost to follow-up. Among the 14 cases that we were able to follow-up, 11 cases had had pain as their primary symptom. Following treatment, this symptom remained unchanged in 1 patient, was improved in 4, and had disappeared in 6; however, there was a recurrence of pain for 3 of these patients. Two patients had sought treatment for cosmetic purposes; following treatment, the lesion disappeared in one and showed a significant reduction in the other. The remaining patient presented with a primary symptom of mouth bleeding, which disappeared following treatment. There were no critical complications. Percutaneous sclerotherapy of venous malformations using polidocanol is safe and effective, and permits repeat treatments. The efficacy is especially good for resolving pain, and complications are minor. It is desirable to use fluoroscopy for these procedures


Evaluating the Effectiveness of an Ultrasonic Acoustic Deterrent for Reducing Bat Fatalities at Wind Turbines.
Large numbers of bats are killed by wind turbines worldwide and minimizing fatalities is critically important to bat conservation and acceptance of wind energy development. We implemented a 2-year study testing the effectiveness of an ultrasonic acoustic deterrent for reducing bat fatalities at a wind energy facility in Pennsylvania. We randomly selected control and treatment turbines that were searched daily in summer and fall 2009 and 2010. Estimates of fatality, corrected for field biases, were compared between treatment and control turbines. In 2009, we estimated 21-51% fewer bats were killed per treatment turbine than per control turbine. In 2010, we determined an approximate 9% inherent difference between treatment and control turbines and when factored into our analysis, variation increased and between 2% more and 64% fewer bats were killed per treatment turbine relative to control turbines. We estimated twice as many hoary bats were killed per control turbine than treatment turbine, and nearly twice as many silver-haired bats in 2009. In 2010, although we estimated nearly twice as many hoary bats and nearly 4 times as many silver-haired bats killed per control turbine than at treatment turbines during the treatment period, these only represented an approximate 20% increase in fatality relative to the pre-treatment period for these species when accounting for inherent differences between turbine sets. Our findings suggest broadband ultrasound broadcasts may reduce bat fatalities by discouraging bats from approaching sound sources. However, effectiveness of ultrasonic deterrents is limited by distance and area ultrasound can be broadcast, in part due to rapid attenuation in humid conditions. We caution that an operational deterrent device is not yet available and further modifications and experimentation are needed. Future efforts must also evaluate cost-effectiveness of deterrents in relation to curtailment strategies to allow a cost-benefit analysis for mitigating bat fatalities.


The emerging role of metformin in gestational diabetes mellitus.
Metformin use during pregnancy is controversial and there is disparity in the acceptance of metformin treatment in women with gestational diabetes mellitus (GDM) in Australia. Despite short term maternal and neonatal safety measures, the placental transfer of metformin during GDM treatment and the absence of long-term safety data in offspring has regulators and prescribers cautious about its use. To determine the current role in GDM management, this literature review describes the physiological changes that occur in GDM and other forms of diabetes in pregnancy (DIP) and international changes in guidelines for GDM diagnosis. Management options are considered, with a focus on the evolving evidence for metformin, its mechanism of action, the maternal, foetal and neonatal outcomes associated with its use and benefit vs risk when compared with the current gold standard, insulin. Investigation reveals a favourable balance of evidence to support the safety and long-term benefits, to mother and child, of using metformin as an alternate to insulin for treatment of GDM. Recent findings of the gastrointestinal-directed action of metformin are at least as important as the hepatic effect and the availability of a novel delayed-release metformin dose form to exploit this new information provides a product and therapeutic strategy ideally suited to the use of metformin in GDM.


Age-Related Oxidative Changes in Primary Porcine Fibroblasts Expressing Mutated Huntingtin.
Huntington's disease (HD) is a devastating neurodegenerative disorder caused by CAG triplet expansions in the huntingtin gene. Oxidative stress is linked to HD pathology, although it is not clear whether this is an effect or a mediator of disease. The transgenic (TgHD) minipig expresses the N-terminal part of human-mutated huntingtin and represents a unique model to investigate therapeutic strategies towards HD. A more detailed characterization of this model is needed to fully utilize its potential. In this study, we focused on the molecular and cellular features of fibroblasts isolated from TgHD minipigs and the wild-type (WT) siblings at different ages, pre-symptomatic at the age of 24-36 months and with the onset of behavioural symptoms at the age of 48 months. We measured oxidative stress, the expression of oxidative stress-related genes, proliferation capacity along with the expression of cyclin B1 and D1 proteins, cellular permeability, and the integrity of the nuclear DNA (nDNA) and mitochondrial DNA in these cells. TgHD fibroblasts isolated from 48-month-old animals showed increased oxidative stress, which correlated with the overexpression of SOD2 encoding mitochondrial superoxide dismutase 2, and the NEIL3 gene encoding DNA glycosylase involved in replication-associated repair of oxidized DNA. TgHD cells displayed an abnormal proliferation capacity and permeability. We further demonstrated increased nDNA damage in pre-symptomatic TgHD fibroblasts (isolated from animals aged 24-36 months). Our results unravel phenotypic alterations in primary fibroblasts isolated from the TgHD minipig model at the age of 48 months. Importantly, nDNA damage appears to precede these phenotypic alterations. Our results highlight the impact of fibroblasts from TgHD minipigs in studying the molecular mechanisms of HD pathophysiology that gradually occur with age.


Epigenetic regulation of microRNAs in acute lymphoblastic leukemia.
To identify microRNAs (miRNAs) epigenetically regulated in acute lymphoblastic leukemia (ALL). We first examined ALL-derived cell lines for the presence of abnormal levels of two different histone modifications (trimethylation of H3 lysine 4 [K4H3me3] and dimethylation of H3 lysine 9 [K9H3me2]) in the 5'UTR regions around CpG islands of 78 miRNAs by chromatin immunoprecipitation (ChIP)-on-ChIP analysis. Methylation status (methylation-specific polymerase chain reaction [PCR]) and expression (quantitative PCR) of miRNAs showing a pattern of histone modifications linked to a closed chromatin structure were analyzed in a panel of six ALL cell lines and in 353 ALL patients. CpG islands around 13 miRNAs disclosed high levels of K9H3me2 and/or low levels of K4H3me3, a pattern of histone modifications underlying a closed chromatin structure associated with repressive gene expression. Complete consistency in the correlation between both histone marks, the presence of DNA methylation around these miRNAs, and their expression patterns was confirmed in the six ALL cell lines. Treatment with 5-Aza-2'-deoxycytidine upregulated the expression levels of these genes, suggesting that epigenetic mechanisms deregulate the expression of these miRNAs. A total of 65% of the ALL samples had at least one miRNA methylated (methylated group). Estimated disease-free survival (DFS) and overall survival (OS) at 14 years were 78% and 71% for nonmethylated patients and 24% and 28% for methylated patients (P = .00001 for both). Multivariate analysis demonstrated that methylation profile was an independent prognostic factor for predicting DFS (P = .0001) and OS (P = .0001). Aberrant miRNA methylation is a common phenomenon in ALL that affects the clinical outcome of these patients.


Associations of a Breast Cancer Polygenic Risk Score With Tumor Characteristics and Survival.
A polygenic risk score (PRS) consisting of 313 common genetic variants (PRS313) is associated with risk of breast cancer and contralateral breast cancer. This study aimed to evaluate the association of the PRS313 with clinicopathologic characteristics of, and survival following, breast cancer. Women with invasive breast cancer were included, 98,397 of European ancestry and 12,920 of Asian ancestry, from the Breast Cancer Association Consortium (BCAC), and 683 women from the European MINDACT trial. Associations between PRS313 and clinicopathologic characteristics, including the 70-gene signature for MINDACT, were evaluated using logistic regression analyses. Associations of PRS313 (continuous, per standard deviation) with overall survival (OS) and breast cancer-specific survival (BCSS) were evaluated with Cox regression, adjusted for clinicopathologic characteristics and treatment. The PRS313 was associated with more favorable tumor characteristics. In BCAC, increasing PRS313 was associated with lower grade, hormone receptor-positive status, and smaller tumor size. In MINDACT, PRS313 was associated with a low risk 70-gene signature. In European women from BCAC, higher PRS313 was associated with better OS and BCSS: hazard ratio (HR) 0.96 (95% CI, 0.94 to 0.97) and 0.96 (95% CI, 0.94 to 0.98), but the association disappeared after adjustment for clinicopathologic characteristics (and treatment): OS HR, 1.01 (95% CI, 0.98 to 1.05) and BCSS HR, 1.02 (95% CI, 0.98 to 1.07). The results in MINDACT and Asian women from BCAC were consistent. An increased PRS313 is associated with favorable tumor characteristics, but is not independently associated with prognosis. Thus, PRS313 has no role in the clinical management of primary breast cancer at the time of diagnosis. Nevertheless, breast cancer mortality rates will be higher for women with higher PRS313 as increasing PRS313 is associated with an increased risk of disease. This information is crucial for modeling effective stratified screening programs.


Apg7p/Cvt2p: A novel protein-activating enzyme essential for autophagy.
In the yeast Saccharomyces cerevisiae, the Apg12p-Apg5p conjugating system is essential for autophagy. Apg7p is required for the conjugation reaction, because Apg12p is unable to form a conjugate with Apg5p in the apg7/cvt2 mutant. Apg7p shows a significant similarity to a ubiquitin-activating enzyme, Uba1p. In this article, we investigated the function of Apg7p as an Apg12p-activating enzyme. Hemagglutinin-tagged Apg12p was coimmunoprecipitated with c-myc-tagged Apg7p. A two-hybrid experiment confirmed the interaction. The coimmunoprecipitation was sensitive to a thiol-reducing reagent. Furthermore, a thioester conjugate of Apg7p was detected in a lysate of cells overexpressing both Apg7p and Apg12p. These results indicated that Apg12p interacts with Apg7p via a thioester bond. Mutational analyses of Apg7p suggested that Cys507 of Apg7p is an active site cysteine and that both the ATP-binding domain and the cysteine residue are essential for the conjugation of Apg7p with Apg12p to form the Apg12p-Apg5p conjugate. Cells expressing mutant Apg7ps, Apg7pG333A, or Apg7pC507A showed defects in autophagy and cytoplasm-to-vacuole targeting of aminopeptidase I. These results indicated that Apg7p functions as a novel protein-activating enzyme necessary for Apg12p-Apg5p conjugation.


Demethylation of H3K27 regulates polycomb recruitment and H2A ubiquitination.
Methylation of histone H3 lysine 27 (H3K27) is a posttranslational modification that is highly correlated with genomic silencing. Here we show that human UTX, a member of the Jumonji C family of proteins, is a di- and trimethyl H3K27 demethylase. UTX occupies the promoters of HOX gene clusters and regulates their transcriptional output by modulating the recruitment of polycomb repressive complex 1 and the monoubiquitination of histone H2A. Moreover, UTX associates with mixed-lineage leukemia (MLL) 2/3 complexes, and during retinoic acid signaling events, the recruitment of the UTX complex to HOX genes results in H3K27 demethylation and a concomitant methylation of H3K4. Our results suggest a concerted mechanism for transcriptional activation in which cycles of H3K4 methylation by MLL2/3 are linked with the demethylation of H3K27 through UTX.


Patients with IDH1 wild type anaplastic astrocytomas exhibit worse prognosis than IDH1-mutated glioblastomas, and IDH1 mutation status accounts for the unfavorable prognostic effect of higher age: implications for classification of gliomas.
WHO grading of human brain tumors extends beyond a strictly histological grading system by providing a basis predictive for the clinical behavior of the respective neoplasm. For example, patients with glioblastoma WHO grade IV usually show a less favorable clinical course and receive more aggressive first-line treatment than patients with anaplastic astrocytoma WHO grade III. Here we provide evidence that the IDH1 status is more prognostic for overall survival than standard histological criteria that differentiate high-grade astrocytomas. We sequenced the isocitrate dehydrogenase 1 gene (IDH1) at codon 132 in 382 patients with anaplastic astrocytoma and glioblastoma from the NOA-04 trial and from a prospective translational cohort study of the German Glioma Network. Patients with anaplastic astrocytomas carried IDH1 mutations in 60%, and patients with glioblastomas in 7.2%. IDH1 was the most prominent single prognostic factor (RR 2.7; 95% CI 1.6-4.5) followed by age, diagnosis and MGMT. The sequence from more favorable to poorer outcome was (1) anaplastic astrocytoma with IDH1 mutation, (2) glioblastoma with IDH1 mutation, (3) anaplastic astrocytoma without IDH1 mutation and (4) glioblastoma without IDH1 mutation (p < 0.0001). In this combined set of anaplastic astrocytomas and glioblastomas both, IDH1 mutation and IDH1 expression status were of greater prognostic relevance than histological diagnosis according to the current WHO classification system. Our data indicate that much of the prognostic significance of patient age is due to the predominant occurrence of IDH1 mutations in younger patients. Immunohistochemistry using a mutation-specific antibody recognizing the R132H mutation yielded similar results. We propose to complement the current WHO classification and grading of high-grade astrocytic gliomas by the IDH1 mutation status and to use this combined histological and molecular classification in future clinical trials.


Reverse Immunology Approach to Define a New HIV-gp41-Neutralizing Epitope.
The design of immunogens susceptible to elicit potent and broadly neutralizing antibodies against the human immunodeficiency virus type 1 (HIV-1) remains a veritable challenge in the course of vaccine development. Viral envelope proteins adopt different conformational states during the entry process, allowing the presentation of transient neutralizing epitopes. We focused on the highly conserved 3S motif of gp41, which is exposed to the surface envelope in its trimeric prefusion state. Vaccination with a W614A-modified 3S peptide induces in animals neutralizing anti-HIV-1 antibodies among which we selected clone F8. We used F8 as bait to select for W614A-3S phage-peptide mimics. Binding and molecular docking studies revealed that F8 interacts similarly with W614A-3S and a Mim_F8-1 mimotope, despite their lack of sequence homology, suggesting structural mimicry. Finally, vaccination of mice with the purified Mim_F8-1 phage elicited HIV-1-neutralizing antibodies that bound to the cognate W614A-3S motif. Collectively, our findings provide new insights into the molecular design of immunogens to elicit antibodies with neutralizing properties.


Do gastrotomies require repair after endoscopic transgastric peritoneoscopy? A controlled study.
The optimal method for closing gastrotomies after transgastric instrumentation has yet to be determined. To compare gastrotomy closure with endoscopically delivered bioabsorbable plugs with no closure. Prospective, controlled study. Animal laboratory. Twenty-three dogs undergoing endoscopic transgastric peritoneoscopy between July and August 2007. Endoscopic anterior wall gastrotomies were performed with balloon dilation to allow passage of the endoscope into the peritoneal cavity. The plug group (n = 12) underwent endoscopic placement of a 4 x 6-cm bioabsorbable mesh plug in the perforation, whereas the no-treatment group (n = 11) did not. Animals underwent necropsy 2 weeks after the procedure. Complications related to gastrotomy closure, gastric burst pressures, relationship of burst perforation to gastrotomy, and the degree of adhesions and inflammation at the gastrotomy site. After the gastrotomy, all dogs survived without any complications. At necropsy, burst pressures were 77 +/- 11 mm Hg and 76 +/- 15 mm Hg (P = .9) in the plug group and no-treatment group, respectively. Perforations occurred at the site of the gastrotomy in 2 of 12 animals in the plug group and in none of the 11 dogs in the no-treatment group (P = .5). Finally, there were minimal adhesions in all dogs (11/11) in the no-treatment group and minimal adhesions in 3 and moderate adhesions or inflammatory masses in 9 of the 12 animals in the plug group (P = .004). Small number of subjects, animal model, no randomization. Gastrotomy trauma during short peritoneoscopy may not be applicable to longer procedures. After endoscopic gastrotomy, animals that were left untreated did not show any clinical ill effects and demonstrated adequate healing, with fewer adhesions and less inflammation compared with those treated with a bioabsorbable plug.


Coarse particles and mortality in three Chinese cities: the China Air Pollution and Health Effects Study (CAPES).
Evidence concerning the health risks of coarse particles (PM(10-2.5)) is limited. There have been no multi-city epidemiologic studies of PM(10-2.5) in developing Asian countries. We examine the short-term association between PM(10-2.5) and daily mortality in three Chinese cities: Beijing, Shanghai, and Shenyang. PM(10-2.5) concentrations were estimated by subtracting PM(2.5) from PM(10) measurements. Data were analyzed using the over-dispersed generalized linear Poisson models. The average daily concentrations of PM(10-2.5) were 101 μg/m(3) for Beijing (2007-2008), 50 μg/m(3) for Shanghai (2004-2008), and 49 μg/m(3) for Shenyang (2006-2008). In the single-pollutant models, the three-city combined analysis showed significant associations between PM(10-2.5) and daily mortality from both total non-accidental causes and from cardiopulmonary diseases. A 10-μg/m(3) increase in 1-day lagged PM(10-2.5) was associated with a 0.25% (95% CI: 0.08 to 0.42) increase in total mortality, 0.25% (95% CI: 0.10 to 0.40) increase in cardiovascular mortality, and 0.48% (95% CI: 0.20 to 0.76) increase in respiratory mortality. However, these associations became statistically insignificant after adjustment for PM(2.5). PM(2.5) was significantly associated with mortality both before and after adjustment for PM(10-2.5). In conclusion, there were no statistically significant associations between PM(10-2.5) and daily mortality after adjustment for PM(2.5) in the three Chinese cities.


Outcome of multimodality treatment of Ewing's sarcoma of the extremities.
The management of Ewing's sarcoma family of tumors (ESFT, Ewing's sarcoma/primitive neuroectodermal tumor) has been established as a multimodality treatment. Advances in imaging and diagnostics, chemotherapy, surgical techniques, radiotherapy and prosthetic technology have resulted in drastic changes in the outcome of this disease, with most of the recent studies having 5-year survival rates of more than 60%. The Indian patients present at a more advanced stage and the compliance of treatment is suboptimal. While there is plenty of data in the world literature on the outcome of Ewing's sarcoma, there is paucity of data in Indian patients. Therefore, we conducted the present study to analyze the outcome of multimodality treatment of ESFT of the extremities at a tertiary nonprofit institute over a decade. 34 patients who had histopathologically proven diagnosis of Ewing's sarcoma of the extremities and had received treatment at our institute from 1997 through 2007 were included for analysis. The majority of patients had involvement of the femur (35%), followed by tibia (17%), fibula and foot (15% each), humerus (12%) and soft tissue of thigh (6%). Twenty-nine patients presented with localized disease (Enneking stage II B) while five patients presented with metastases (Enneking stage III). All patients received Vincristine, Actinomycin D, Cyclofosfamide + Ifosfamide and Etoposide (VAC+IE)-based chemotherapy and local treatment was offered to all but three patients having multicentric disease. The local treatment offered were, radiation (n= 15), surgery (n= 12) both surgery and radiation (n=4). All patients were analyzed for oncological outcome (event-free and overall survival, local and systemic relapses) by clinical and imaging evaluation and functional outcome by using the musculoskeletal tumor society (MSTS) score. These outcomes were correlated with age, sex, size of tumor, stage at presentation, modality of local treatment and site of relapse. At the final follow-up (mean, 26 months; median, 17 months; range, 3-97 months), the overall and event-free survivals were 47 ± 12% and 34 ± 9%, respectively. Sixty-two percent of the patients presented with a tumor size more than 8 cm. On correlation with age, sex, size of tumor, stage at presentation, modality of local treatment and site of relapse, no correlation of survival was seen with any of the variables except event-free survival with size of the tumor. The functional outcome of all the patients was satisfactory (MSTS score >16 out of 30). No patient underwent amputation. Although the demographic profile, stage at presentation and the local and systemic treatment regimen followed in our study was similar to the world literature, the outcome of Ewing's sarcoma in Indian patients were found to be inferior to that reported in the western literature. Larger multicentric studies with longer follow-up are required to exactly determine the key areas crucial in improving this outcome.


The Gynecologic Health Consequences of Chlamydia trachomatis Infection in Military Servicewomen.
Chlamydia trachomatis is the most common sexually transmitted bacterial infection in the United States. Within the U.S. military, the age- and race-adjusted chlamydia infection rates among female service members are consistently higher than civilian rates, with a 20% annual acquisition rate among young active-duty women. The sequelae of chlamydia disproportionately impact women in terms of severity and cost. Untreated chlamydia progresses to pelvic inflammatory disease in 40% of cases, and is a leading cause of fallopian tube damage and pelvic adhesive disease resulting in ectopic pregnancy, tubal infertility, and acute and chronic pelvic pain. Tubal infertility is among the leading indications for in vitro fertilization (IVF) nationally and rates among couples undergoing IVF at military treatment centers are double the national average. Collectively, chlamydia infection represents a significant resource burden to the military health care system and, in view of the serious gynecologic health sequelae, a significant threat to the readiness of servicewomen. In this review, we discuss the gynecologic impact of chlamydia infection within the military, the critical gaps for research funding, and opportunities for intervention.


Effects of lithium on platelet ionic intracellular calcium concentration in patients with bipolar (manic-depressive) disorder and healthy controls.
Lithium is an effective drug in the treatment of both manic and depressive episodes of bipolar disorder. Lithium has been shown to block the metabolism of the intracellular second messenger inositol-1,4,5-trisphosphate which is involved in the rise in ionic intracellular calcium [( Ca++]i) which triggers neurotransmitter release and other cellular changes in secretory cells. We have measured the effect of lithium on [Ca++]i dynamics in platelets from bipolar patients stabilized with lithium treatment, and from healthy controls. Both resting [Ca++]i and the thrombin stimulated increase in [Ca++]i were higher in bipolar patients than in controls. Lithium added in vitro tended to increase the thrombin-stimulated rise in [Ca++]i. The use of the fluorescent Ca++ probe fura-2 in human platelets provides a useful method to investigate the mechanism of lithium's action in bipolar disorder and to study Ca++ related systems which may be abnormal in bipolar disorders.


Comparative phenotypic and functional analyses of the effects of autologous plasma and recombinant human macrophage-colony stimulating factor (M-CSF) on porcine monocyte to macrophage differentiation.
Porcine monocyte-derived macrophages (moMΦ) have been employed as a model cell in numerous studies of the porcine immune system. However, the lack of a standardized method for moMΦ differentiation hampers the comparison of results coming from the use of different laboratory protocols. In this study we compared the use of varying concentrations of autologous plasma (10, 20 and 30% v/v) or recombinant human macrophage-colony stimulating factor (hM-CSF; 50, 100, and 200ng/ml) to differentiate porcine monocytes into macrophages. Changes in cell morphology and surface marker expression were assessed by confocal microscopy and flow cytometry. Macrophage differentiation was evaluated by analysing TNF-α response to LPS stimulation and determining cytokine secretion patterns under both basal conditions and after classical and alternative activation. The effects of the differentiation methods on metabolic activity and susceptibility to infection with the myelotropic African swine fever virus (ASFV) were also evaluated. Monocytes cultured using the different culture conditions tested augmented in dimension and cellular complexity, but increasing porcine plasma concentrations resulted in a dose dependent enhancement in granularity and a marked pleomorphism. As expected, CD163, MHC class II DR and CD203a expression were up-regulated in both hM-CSF (M-CSF-moMΦ) and autologous plasma cultured macrophages (AP-moMΦ), although a lower percentage of CD163+ cells were found following differentiation with high percentages of porcine plasma. We observed enhanced number of viable cells using high concentration of hM-CSF compared to porcine plasma, suggesting a proliferative effect. Irrespective of differentiation conditions, monocyte differentiation into macrophages resulted in an increased susceptibility to ASFV and yielded larger amounts of LPS-induced TNF-α. AP-moMΦ showed a higher basal release of IL-1RA compared to those cultured with hM-CSF and displayed a reduced ability to respond to classical activation, suggesting that the use of high percentages of porcine plasma led to the acquisition of a M2-like phenotype. We conclude that all the protocols tested in this study can be considered as suitable to produce porcine moMΦ, although the use of hM-CSF provides high responsiveness to M1 polarization. Since a higher phenotypic and functional inter-animal variability was observed in AP-moMΦ, we propose that the use of low concentration of hM-CSF should be adopted as the method of choice to provide a better reproducibility between experiments.