Ultrasonographic features of superficial and nodular basal cell carcinoma.
To describe the ultrasonographic findings of surface and nodular basal cell skin cancer (BCC) using high frequency ultrasonography. We examined 60 primary BCCs in different locations with the High Frequency Ultrasound (HFU) system DUB Skin Scanner using 75 MHz and 30 MHz probes. Epidermis, dermis, and depth of tumors spread in the region of interest (ROI) were measured. Visually unchanged, contralateral skin areas were examined as the control. Results: The surface BCC most often had elongated contours, clear margins and hypoechoic structure, while the nodular BCC had round or oval outlines and diffusely hypo-heterogeneous structure with clear margins. Sclerodermiform BCCs were visualized as hypoechoic areas of irregular shape penetrating in the dermis, with wavy fuzzy margins. The average thickness of the surface BCC in the US examination was 556.28±136.95 μ, the nodular BCC thickness was 2439.71±865.92 μ and the sclerodermiform thickness was 1500±325.33 μ. A statistically significant increase in the average thickness of tumors of the nodularand scleroderma forms was observed in comparison with the surface clinical variant (p<0.05). Hyperechoic inclusions were observed in 11% of the surface BCC's and in the 100% of the nodular BCC's. Their average number was 2±0.57 and 4±4.8, with the average area of 0.03±0.02 mm2 and 0.04±0.03 mm2 (p>0.05), respectively. In the surface BCC, they were mainly located along the periphery of the hypoechoic zones. In nodular BCC, the inclusions had a peripheral and combined (center and peripheral) distribution. Ultrasound allows differentiating BCC as diffuse-heterogeneous, hypoechoic, formations in the dermis with distinct contours. Depending on the clinical picture, they differ in form, depth of bedding, as well as in the quantitative ratio and distribution of the point hyperechoic structures in them.


Osteopathic manipulative treatment for inpatients with pulmonary exacerbations of cystic fibrosis: effects on spirometry findings and patient assessments of breathing, anxiety, and pain.
Osteopathic manipulative treatment (OMT) has been studied in patients with various respiratory diseases. However, to the authors' knowledge, no studies have assessed the efficacy of OMT in patients with cystic fibrosis (CF). To evaluate pulmonary function and perceptions of breathing, anxiety, and pain of CF patients who receive OMT in addition to standard inpatient management of pulmonary exacerbation. In a single-blind randomized controlled trial, we assessed adult patients with a history of CF who were admitted to the hospital because of pulmonary exacerbation. Participants were randomly assigned to receive a daily standardized protocol of OMT or sham therapy. Both groups also received standard treatment for CF. Spirometry and questionnaire data (self-assessment of breathing, pain, and anxiety level) were collected before the first OMT or sham therapy session and after the final session. A total of 33 patients were included in the study: 16 in the OMT group and 17 in the sham therapy group. Improvements in spirometric parameters were observed in both the OMT and the sham therapy groups, with no statistically significant differences found between the groups. More patients in the OMT group than in the sham therapy group had questionnaire response patterns that indicated their breathing had improved during the study period (15 of 16 vs 8 of 16, respectively). No differences were found between groups for perceived improvement of pain and anxiety. In the current study, CF patients who received OMT did not demonstrate statistically significant differences in pre- and posttreatment spirometry findings compared with CF patients who received sham therapy. Questionnaire findings suggest that OMT may affect CF patients' perception of overall quality of breathing. Additional studies are needed to assess the clinical use of OMT in patients with CF.


Social network support and harm reduction activities in a peer researcher-led pilot study, British Columbia, Canada.
People who smoke drugs (PWSD) are at high risk of both infectious disease and overdose. Harm reduction activities organized by their peers in the community can reduce risk by providing education, safer smoking supplies, and facilitate access to other services. Peers also provide a network of people who provide social support to PWSD which may reinforce harm reducing behaviors. We evaluated the numbers of supportive network members and the relationships between received support and participants' harm-reducing activities. Initial peer-researchers with past or current lived drug use experience were employed from communities in Abbotsford and Vancouver to interview ten friends from their social networks who use illegal drugs mainly through smoking. Contacts completed a questionnaire about people in their own harm reduction networks and their relationships with each other. We categorized social support into informational, emotional, and tangible aspects, and harm reduction into being trained in the use of, or carrying naloxone, assisting peers with overdoses, using brass screens to smoke, obtaining pipes from service organizations and being trained in CPR. Fifteen initial peer researchers interviewed 149 participants who provided information on up to 10 people who were friends or contacts and the relationships between them. People who smoked drugs in public were 1.46 (95% CI, 1.13-1.78) more likely to assist others with possible overdoses if they received tangible support; women who received tangible support were 1.24 (95% CI; 1.02-1.45) more likely to carry and be trained in the use of naloxone. There was no relationship between number of supportive network members and harm reduction behaviors. In this pilot study, PWSD who received tangible support were more likely to assist peers in possible overdoses and be trained in the use of and/or carry naloxone, than those who did not receive tangible support. Future work on the social relationships of PWSD may prove valuable in the search for credible and effective interventions.


Polymeric Nanoparticles' Accumulation in Atopic Dermatitis: Clinical Comparison between Healthy, Non-Lesional, and Lesional Skin.
A major limitation in the current topical treatment strategies for inflammatory skin disorders is the inability to selectively target the inflamed site with minimal exposure of healthy skin. Atopic dermatitis is one of the most prevalent types of dermatitis. The use of polymeric nanoparticles for targeting inflamed skin has been recently proposed, and therefore the aim of this proof-of-concept clinical study was to investigate the skin penetration and deposition of polymeric biodegradable nanoparticles in the atopic dermatitis lesions and compare the data obtained to the deposition of the particles into the healthy skin or lesion-free skin of the atopic dermatitis patients. For that, fluorescent PLGA nanoparticles in sizes of approximately 100 nm were prepared and applied to the skin of healthy volunteers and the lesional and non-lesional skin of atopic dermatitis patients. Skin biopsies were examined using confocal laser scanning microscopy to track the skin deposition and depth of penetration of the particles. Immunohistochemistry was performed to investigate the alteration in tight-junction protein distribution in the different types of skin. Results have shown that nanoparticles were found to have higher deposition into the atopic dermatitis lesions with minimal accumulation in healthy or non-lesional skin. This has been primarily correlated with the impaired barrier properties of atopic dermatitis lesions with the reduced production of Claudin-1. It was concluded that polymeric nanoparticles offer a potential tool for selective drug delivery to inflamed skin with minimal exposure risk to healthy skin.


Zinc finger nuclease and homing endonuclease-mediated assembly of multigene plant transformation vectors.
Binary vectors are an indispensable component of modern Agrobacterium tumefaciens-mediated plant genetic transformation systems. A remarkable variety of binary plasmids have been developed to support the cloning and transfer of foreign genes into plant cells. The majority of these systems, however, are limited to the cloning and transfer of just a single gene of interest. Thus, plant biologists and biotechnologists face a major obstacle when planning the introduction of multigene traits into transgenic plants. Here, we describe the assembly of multitransgene binary vectors by using a combination of engineered zinc finger nucleases (ZFNs) and homing endonucleases. Our system is composed of a modified binary vector that has been engineered to carry an array of unique recognition sites for ZFNs and homing endonucleases and a family of modular satellite vectors. By combining the use of designed ZFNs and commercial restriction enzymes, multiple plant expression cassettes were sequentially cloned into the acceptor binary vector. Using this system, we produced binary vectors that carried up to nine genes. Arabidopsis (Arabidopsis thaliana) protoplasts and plants were transiently and stably transformed, respectively, by several multigene constructs, and the expression of the transformed genes was monitored across several generations. Because ZFNs can potentially be engineered to digest a wide variety of target sequences, our system allows overcoming the problem of the very limited number of commercial homing endonucleases. Thus, users of our system can enjoy a rich resource of plasmids that can be easily adapted to their various needs, and since our cloning system is based on ZFN and homing endonucleases, it may be possible to reconstruct other types of binary vectors and adapt our vectors for cloning on multigene vector systems in various binary plasmids.


2q36.3 is associated with prognosis for oestrogen receptor-negative breast cancer patients treated with chemotherapy.
Large population-based registry studies have shown that breast cancer prognosis is inherited. Here we analyse single-nucleotide polymorphisms (SNPs) of genes implicated in human immunology and inflammation as candidates for prognostic markers of breast cancer survival involving 1,804 oestrogen receptor (ER)-negative patients treated with chemotherapy (279 events) from 14 European studies in a prior large-scale genotyping experiment, which is part of the Collaborative Oncological Gene-environment Study (COGS) initiative. We carry out replication using Asian COGS samples (n=522, 53 events) and the Prospective Study of Outcomes in Sporadic versus Hereditary breast cancer (POSH) study (n=315, 108 events). Rs4458204_A near CCL20 (2q36.3) is found to be associated with breast cancer-specific death at a genome-wide significant level (n=2,641, 440 events, combined allelic hazard ratio (HR)=1.81 (1.49-2.19); P for trend=1.90 × 10(-9)). Such survival-associated variants can represent ideal targets for tailored therapeutics, and may also enhance our current prognostic prediction capabilities.


Logistic random effects regression models: a comparison of statistical packages for binary and ordinal outcomes.
Logistic random effects models are a popular tool to analyze multilevel also called hierarchical data with a binary or ordinal outcome. Here, we aim to compare different statistical software implementations of these models. We used individual patient data from 8509 patients in 231 centers with moderate and severe Traumatic Brain Injury (TBI) enrolled in eight Randomized Controlled Trials (RCTs) and three observational studies. We fitted logistic random effects regression models with the 5-point Glasgow Outcome Scale (GOS) as outcome, both dichotomized as well as ordinal, with center and/or trial as random effects, and as covariates age, motor score, pupil reactivity or trial. We then compared the implementations of frequentist and Bayesian methods to estimate the fixed and random effects. Frequentist approaches included R (lme4), Stata (GLLAMM), SAS (GLIMMIX and NLMIXED), MLwiN ([R]IGLS) and MIXOR, Bayesian approaches included WinBUGS, MLwiN (MCMC), R package MCMCglmm and SAS experimental procedure MCMC.Three data sets (the full data set and two sub-datasets) were analysed using basically two logistic random effects models with either one random effect for the center or two random effects for center and trial. For the ordinal outcome in the full data set also a proportional odds model with a random center effect was fitted. The packages gave similar parameter estimates for both the fixed and random effects and for the binary (and ordinal) models for the main study and when based on a relatively large number of level-1 (patient level) data compared to the number of level-2 (hospital level) data. However, when based on relatively sparse data set, i.e. when the numbers of level-1 and level-2 data units were about the same, the frequentist and Bayesian approaches showed somewhat different results. The software implementations differ considerably in flexibility, computation time, and usability. There are also differences in the availability of additional tools for model evaluation, such as diagnostic plots. The experimental SAS (version 9.2) procedure MCMC appeared to be inefficient. On relatively large data sets, the different software implementations of logistic random effects regression models produced similar results. Thus, for a large data set there seems to be no explicit preference (of course if there is no preference from a philosophical point of view) for either a frequentist or Bayesian approach (if based on vague priors). The choice for a particular implementation may largely depend on the desired flexibility, and the usability of the package. For small data sets the random effects variances are difficult to estimate. In the frequentist approaches the MLE of this variance was often estimated zero with a standard error that is either zero or could not be determined, while for Bayesian methods the estimates could depend on the chosen "non-informative" prior of the variance parameter. The starting value for the variance parameter may be also critical for the convergence of the Markov chain.


Increased hTR expression during transition from adenoma to carcinoma is not associated with promoter methylation.
Human telomerase RNA component (hTR) expression, which increases in the majority of cancer cells with an acquisition of telomerase activity, was concomitantly evaluated with methylation status and human telomerase reverse transcriptase (hTERT) expression in colorectal cancers and precursor lesions. hTR and hTERT expressions were detected by in situ hybridization and reverse transcription following polymerase chain reaction, respectively, in 15 colonic adenomas, 19 sporadic colonic cancers at various histological stages, and 3 normal colonic mucosa samples. The methylationstatus of hTR was evaluated by methylation-specific polymerase chain reaction following restriction endonuclease digestion and direct sequencing. hTERT expression was detected in 16 of 19 cancers. hTR expression was detected in all cancers including two cases of intramucosal carcinoma. No hTR signals were detected in the normal epithelium or in the adenomas with severe atypism. CpG dinucleotides in the 5'-untranslated region of hTR were completely unmethylated from -204 to -3 and mosaically methylated from -290 to -272, irrespective of the atypism. These results suggest that hTR expression is increased at the adenoma-to-carcinoma transition stage but is not always associated with hTERT expression. Hypomethylation of the hTR promoter region is not likely to be the main mechanism regulating hTR expression.


Application of CBNAAT (Xpert MTB/RIF assay) in new smear negative pulmonary tuberculosis patients.
Presumptive pulmonary tuberculosis (PTB) patients whose sputum are detected to be smear negative for acid fast bacilli (AFB) present a significant challenge for a treating physician. Initiating these patients on anti tuberculous treatment (ATT) on empirical basis is not a good strategy as many were found to be sputum culture for tuberculosis negative on further evaluation according to many previous studies. In India due to resource limited settings and lack of knowledge about newest diagnostic modalities patients are often initiated only on the basis of characteristic clinical symptoms and chest radiographic abnormalities. This study was conducted to identify the advantage of application of sputum cartridge based nucleic acid amplification test (CBNAAT) in sputum AFB smear negative presumptive pulmonary TB patients. Our study concluded that clinical symptoms and radiological characteristics cannot differentiate TB patients from non-TB patients. Treating patients only on empirical basis would have resulted in unnecessary treatment of 41 patients.


Phase 2 clinical trial of PBI-4050 in patients with idiopathic pulmonary fibrosis.
PBI-4050 is a novel orally active small-molecule compound with demonstrated anti-fibrotic activity in several models of fibrosis, including lung fibrosis. We present results from our first clinical study of PBI-4050 in patients with idiopathic pulmonary fibrosis (IPF).This 12-week open-label study explored the safety, efficacy and pharmacokinetics of daily oral doses of 800 mg PBI-4050 alone and in combination with nintedanib or pirfenidone in patients with predominantly mild or moderate IPF. Nine patients received PBI-4050 alone, 16 patients received PBI-4050 with nintedanib and 16 patients received PBI-4050 with pirfenidone.PBI-4050 alone or in combination with nintedanib or pirfenidone was well tolerated. Pharmacokinetic profiles for PBI-4050 were similar in the PBI-4050 alone and PBI-4050+nintedanib groups but reduced in the PBI-4050+pirfenidone group, suggesting a drug-drug interaction. There were no significant changes in forced vital capacity (FVC), either in % predicted or mL, from baseline to week 12 for PBI-4050 alone or PBI-4050+nintedanib. In contrast, a statistically significant reduction (p<0.024) in FVC % pred was seen for PBI-4050+pirfenidone after 12 weeks.There were no safety concerns with PBI-4050 alone or in combination with nintedanib or pirfenidone in IPF patients. The stability of FVC between baseline and week 12 looked encouraging for PBI-4050 alone and in combination with nintedanib.


Spanning the scales of granular materials through microscopic force imaging.
If you walk on sand, it supports your weight. How do the disordered forces between particles in sand organize, to keep you from sinking? This simple question is surprisingly difficult to answer experimentally: measuring forces in three dimensions, between deeply buried grains, is challenging. Here we describe experiments in which we have succeeded in measuring forces inside a granular packing subject to controlled deformations. We connect the measured micro-scale forces to the macro-scale packing force response with an averaging, mean field calculation. This calculation explains how the combination of packing structure and contact deformations produce the observed nontrivial mechanical response of the packing, revealing a surprising microscopic particle deformation enhancement mechanism.


Combined therapies in the treatment of neurotrauma: polymers, bridges and gene therapy in visual system repair.
The mature central nervous system (CNS) has limited capacity for self-renewal and repair after injury or neurodegeneration, and therapeutic strategies are needed to promote the viability of damaged neurons and the regrowth of their axons. The retina and optic nerve (ON) are part of the CNS, and the visual system is widely used in experimental studies on injury and repair. To test various cellular and molecular approaches in attempts to replace retinal ganglion cells (RGCs) in depleted retinas or, more usually, promote the survival of endogenous injured RGCs and stimulate axonal regeneration after ON or intracranial optic tract (OT) injury. Intraocular injections of brain-derived neurotrophic factor and ciliary neurotrophic factor (CNTF) temporarily increase RGC survival after ON injury. More sustained neuroprotection is obtained using adeno-associated viral vectors to transfect RGCs with brain-derived neurotrophic factor or CNTF genes. After ON crush, intravitreal adeno-associated viral CNTF injections also increase RGC axonal regrowth. Additional protective and growth effects are obtained after intraocular elevation of cAMP and by manipulation of protein kinase signalling pathways in RGCs. Regeneration is increased by transplanting a segment of peripheral nerve onto the cut ON. Schwann cells in peripheral nerve grafts can be genetically modified using lentiviral vectors to over-express CNTF, resulting in increased regrowth of RGC axons. After OT lesions, hydrogels have been used to bridge the injury, sometimes with the incorporation of signalling peptides or cells genetically modified to express neurotrophic factors. There is now a general consensus that combinatorial approaches are needed to elicit sustained and effective regenerative responses in injured adult CNS neurons.


Clearance of the mutant androgen receptor in motoneuronal models of spinal and bulbar muscular atrophy.
Spinal and bulbar muscular atrophy (SBMA) is an X-linked motoneuron disease caused by an abnormal expansion of a tandem CAG repeat in exon 1 of the androgen receptor (AR) gene that results in an abnormally long polyglutamine tract (polyQ) in the AR protein. As a result, the mutant AR (ARpolyQ) misfolds, forming cytoplasmic and nuclear aggregates in the affected neurons. Neurotoxicity only appears to be associated with the formation of nuclear aggregates. Thus, improved ARpolyQ cytoplasmic clearance, which indirectly decreases ARpolyQ nuclear accumulation, has beneficial effects on affected motoneurons. In addition, increased ARpolyQ clearance contributes to maintenance of motoneuron proteostasis and viability, preventing the blockage of the proteasome and autophagy pathways that might play a role in the neuropathy in SBMA. The expression of heat shock protein B8 (HspB8), a member of the small heat shock protein family, is highly induced in surviving motoneurons of patients affected by motoneuron diseases, where it seems to participate in the stress response aimed at cell protection. We report here that HspB8 facilitates the autophagic removal of misfolded aggregating species of ARpolyQ. In addition, though HspB8 does not influence p62 and LC3 (two key autophagic molecules) expression, it does prevent p62 bodies formation, and restores the normal autophagic flux in these cells. Interestingly, trehalose, a well-known autophagy stimulator, induces HspB8 expression, suggesting that HspB8 might act as one of the molecular mediators of the proautophagic activity of trehalose. Collectively, these data support the hypothesis that treatments aimed at restoring a normal autophagic flux that result in the more efficient clearance of mutant ARpolyQ might produce beneficial effects in SBMA patients.


The Role of Temporal Acoustic Exaggeration in High Variability Phonetic Training: A Behavioral and ERP Study.
High variability phonetic training (HVPT) has been found to be effective in helping adult learners acquire non-native phonetic contrasts. The present study investigated the role of temporal acoustic exaggeration by comparing the canonical HVPT paradigm without involving acoustic exaggeration with a modified adaptive HVPT paradigm that integrated key temporal exaggerations in infant-directed speech (IDS). Sixty native Chinese adults participated in the training of the English /i/ and /i/ vowel contrast and were randomly assigned to three subject groups. Twenty were trained with the typical HVPT paradigm (the HVPT group), twenty were trained under the modified adaptive approach with acoustic exaggeration (the HVPT-E group), and twenty were in the control group. Behavioral tasks for the pre- and post- tests used natural word identification, synthetic stimuli identification, and synthetic stimuli discrimination. Mismatch negativity (MMN) responses from the HVPT-E group were also obtained to assess the training effects in within- and across- category discrimination without requiring focused attention. Like previous studies, significant generalization effects to new talkers were found in both the HVPT group and the HVPT-E group. The HVPT-E group, by contrast, showed greater improvement as reflected in larger progress in natural word identification performance. Furthermore, the HVPT-E group exhibited more native-like categorical perception based on spectral cues after training, together with corresponding training-induced changes in the MMN responses to within- and across- category differences. These data provide the initial evidence supporting the important role of temporal acoustic exaggeration with adaptive training in facilitating phonetic learning and promoting brain plasticity at the perceptual and pre-attentive neural levels.


Detection of polycyclic aromatic hydrocarbons from cooking oil using ultra-thin layer chromatography and surface enhanced Raman spectroscopy.
The combinatory use of on-chip ultra-thin layer chromatography (UTLC) and surface enhanced Raman spectroscopy (SERS) has been demonstrated to detect mixtures of hazardous food contaminants, polycyclic aromatic hydrocarbons (PAHs), from cooking oil samples. After a simple acetonitrile extraction step, the organic phase was directly applied to the chemically modified SERS-active silver nanorod (AgNR) substrate without further cleanup, and subjected to UTLC separation. The spectral interference from co-extracted oil residues was mitigated by UTLC, and the SERS detection limits were found to be equivalent to or lower than those found in standard PAH solutions. The interference from the oil matrix was also quantitatively assessed, and the PAH extraction procedure was optimized for UTLC-SERS. This UTLC-SERS strategy provides a simple but effective method for on-chip sample cleanup and sensing from complex sample matrices.


Tuberculosis in kindergarten and primary school, Italy, 2008-2009.
An outbreak of tuberculosis (TB) in Italy involved 19 schoolchildren with active TB and 43 with latent infection. The source of the outbreak was a school assistant born in Italy who had a family history of TB. This outbreak highlights the need for maintaining clinical and public health expertise in countries with low TB incidence.


Specific humoral immune response induced by propionibacterium acnes can prevent Actinobacillus pleuropneumoniae infection in mice.
Porcine contagious pleuropneumonia, caused by Actinobacillus pleuropneumoniae, has a major impact on economics, ecology, and animal welfare in the pig-rearing industry. Propionibacterium acnes, a facultative anaerobic Gram-positive corynebacterium, exists widely in normal healthy adult animals. We have shown previously that P. acnes can prevent A. pleuropneumoniae infections in mice and pigs. To elucidate the mechanism of this effect and to identify novel A. pleuropneumoniae vaccines, the role of anti-P. acnes antibodies in preventing infection was analyzed by indirect immunofluorescence and opsonophagocytosis assays in vitro. The role of the specific humoral immune response induced by P. acnes was confirmed in a B cell depletion mouse model. The survival rates of mice challenged with A. pleuropneumoniae exhibited a highly significant positive rank correlation with the levels of anti-P. acnes antibodies. The specific antibodies induced by P. acnes had the ability to combine with A. pleuropneumoniae and increase opsonization of A. pleuropneumoniae for phagocytosis. Furthermore, analysis in the murine B cell depletion model confirmed that the humoral immune response induced by P. acnes played an important role in resistance to A. pleuropneumoniae infection. In this study, we further elucidated the reasons that P. acnes can prevent A. pleuropneumoniae infection, which provides useful evidence for the development of heterologous vaccines for the control of porcine contagious pleuropneumonia.


Reference-unbiased copy number variant analysis using CGH microarrays.
Comparative genomic hybridization (CGH) microarrays have been used to determine copy number variations (CNVs) and their effects on complex diseases. Detection of absolute CNVs independent of genomic variants of an arbitrary reference sample has been a critical issue in CGH array experiments. Whole genome analysis using massively parallel sequencing with multiple ultra-high resolution CGH arrays provides an opportunity to catalog highly accurate genomic variants of the reference DNA (NA10851). Using information on variants, we developed a new method, the CGH array reference-free algorithm (CARA), which can determine reference-unbiased absolute CNVs from any CGH array platform. The algorithm enables the removal and rescue of false positive and false negative CNVs, respectively, which appear due to the effects of genomic variants of the reference sample in raw CGH array experiments. We found that the CARA remarkably enhanced the accuracy of CGH array in determining absolute CNVs. Our method thus provides a new approach to interpret CGH array data for personalized medicine.


Nitrous Oxide Is a Potent Inhibitor of Bacterial Reductive Dechlorination.
Organohalide-respiring bacteria are key players for the turnover of organohalogens. At sites impacted with chlorinated ethenes, bioremediation promotes reductive dechlorination; however, stoichiometric conversion to environmentally benign ethene is not always achieved. We demonstrate that nitrous oxide (N2O), a compound commonly present in groundwater, inhibits organohalide respiration. N2O concentrations in the low micromolar range decreased dechlorination rates and resulted in incomplete dechlorination of tetrachloroethene (PCE) in Geobacter lovleyi strain SZ and of cis-1,2-dichloroethene ( cDCE) and vinyl chloride (VC) in Dehalococcoides mccartyi strain BAV1 axenic cultures. Presumably, N2O interferes with reductive dechlorination by reacting with super-reduced Co(I)-corrinoids of reductive dehalogenases, which is supported by the finding that N2O did not inhibit corrinoid-independent fumarate-to-succinate reduction in strain SZ. Kinetic analyses revealed a best fit to the noncompetitive Michaelis-Menten inhibition model and determined N2O inhibitory constants, KI, for PCE and cDCE dechlorination of 40.8 ± 3.8 and 21.2 ± 3.5 μM in strain SZ and strain BAV1, respectively. The lowest KI value of 9.6 ± 0.4 μM was determined for VC to ethene reductive dechlorination in strain BAV1, suggesting that this crucial dechlorination step for achieving detoxification is most susceptible to N2O inhibition. Groundwater N2O concentrations exceeding 100 μM are not uncommon, especially in watersheds impacted by nitrate runoff from agricultural sources. Thus, dissolved N2O measurements can inform about cDCE and VC stalls at sites impacted with chlorinated ethenes.


[Using the SETQ system to evaluate and improve teaching qualities of clinical teachers].
To determine (a) the feasibility of implementing a system for the evaluation of teaching qualities (SETQ) of faculty in an academic medical centre, (b) the psychometric qualities of the questionnaires that are used for the generation of feedback and (c) how residents evaluate the teaching qualities of faculty members and how faculty rated themselves. Questionnaire study. Residents evaluated the teaching qualities of faculty members and faculty also evaluated themselves. Specialty specific questionnaires were developed for both evaluations. The psychometric qualities of the questionnaires were determined by using exploratory factor analysis and by calculating the reliability coefficients of scale constructs and item-total correlation. Mean, median and range were calculated for all teaching aspects per training programme. In the course of one year, 16 residency programmes in our academic medical centre implemented the SETQ system for the evaluation of teaching faculty. 398 faculty members and 314 residents were invited to (self-) evaluate; the response rates were 80.9% and 73.6% respectively. Residents conducted 2,520 evaluations in total. Factor analysis resulted in the definition of 5 teaching domains: 'learning climate', 'professional attitude towards residents', 'communication of learning goals', 'evaluation of residents' and 'feedback to residents'. Item-total correlation and reliability were high for both the residents' questionnaire and the self-evaluation questionnaire (Cronbach's alpha: > 0.70), except for the educational aspect 'learning climate' on the self-evaluation questionnaire (alpha: 0.67) Faculty members were rated positively, but the self-evaluation of faculty members was slightly less positive. For both groups 'professional attitude towards residents' was the highest scoring teaching domain and 'communication of learning goals' the lowest one. Implementing a system for the evaluation of teaching qualities (SETQ) of faculty in an academic hospital was proven feasible. The psychometric qualities of the underlying instruments was sufficient to good. Teaching faculty were evaluated positively, although interdepartmental variations existed.


Hepatitis C virus infection and liver steatosis.
The mechanism by which the hepatitis C virus (HCV) causes chronic, progressive liver damage is unknown. Factors other than the virus itself have been implicated. The role of liver steatosis has been recently studied. Hepatic steatosis is a common histological finding occurring in more 50% of patients with chronic hepatitis C. Both host and viral factors have been demonstrated to play an important role in its development. In those patients infected with genotype 1, steatosis appears to be due to the co-existence of Non-Alchoholic SteatoHepatitis (NASH) with HCV and associated with an increased body mass index (BMI). Some recent observations suggest that steatosis may be of viral origin and related to genotype 3. This fact raises the possibility of a direct effect of specific viral sequences on the pathogenesis of lipid accumulation. Furthermore, hepatic steatosis attributed to genotype 3 correlates directly with serum and intrahepatic titters of HCV RNA. The resolution of steatosis after successful antiviral therapy as well as steatosis being a sign of recurrent HCV infection in patients with genotype 3 add convincing evidence that steatosis is viral related. The pathogenic mechanism induced by genotype 3 is speculative. A correlation between steatosis, intrahepatic HCV RNA and core protein expression suggest a direct effect. Further support is provided by the finding that HCV core protein induces steatosis in transgenic mice. Another possibility relates to interaction with hepatic triglyceride turnover. In conclusion, for patients infected with genotype 1, BMI has a role in the pathogenesis of steatosis while in those infected with genotype 3, steatosis may be due to a virus-specific cytopathic effect. Regardless of etiology, the contribution of both to liver fibrosis progression seems accepted.


Guidelines for the management of haemodynamically stable patients with stab wounds to the anterior abdomen.
Clinical practice guidelines have been shown to improve the delivery of care. Anterior abdominal stab wounds, although uncommon, pose a challenge in both rural and urban trauma care. A multidisciplinary working party was established to assist in the development of evidence-based guidelines to answer three key clinical questions: (i) What is the ideal prehospital management of anterior abdominal stab wounds? (ii) What is the ideal management of anterior abdominal stab wounds in a rural or urban hospital without an on-call surgeon? (iii) What is the ideal emergency management of stable patients with anterior abdominal stab wounds when surgical service is available? A systematic review, using Cochrane method, was undertaken. The data were graded by level of evidence as outlined by the Australian National Health and Medical Research Council. Stable patients with anterior abdominal stab wounds should be transported to the hospital without delay. Any interventions deemed necessary in prehospital care should be undertaken en route to hospital. In rural hospitals with no on-call surgeon, local wound exploration (LWE) may be undertaken by a general practitioner if confident in this procedure. Otherwise or in the presence of obvious fascial penetration, such as evisceration, the patient should be transferred to the nearest main trauma service for further management. In urban hospitals the patient with omental or bowel evisceration or generalized peritonitis should undergo urgent exploratory laparotomy. Stable patients may be screened using LWE. Abdominal computed tomography scan and plain radiographs are not indicated. Obese and/or uncooperative patients require a general anaesthetic for laparoscopy. If there is fascial penetration on LWE or peritoneal penetration on laparoscopy, then an urgent laparotomy should be undertaken. The developed evidence-based guidelines for stable patients with anterior abdominal stab wounds may help minimize unnecessary diagnostic tests and non-therapeutic laparotomy rates.


Ten years of experience with third and fourth branchial remnants.
Third and fourth branchial remnants may result in cysts and abscesses that are in close contact with the thyroid gland. These anomalies are rare and often present diagnostic and therapeutic challenges. The charts of patients diagnosed with a branchial anomaly between July 1991 and July 2001 at the Montreal Children's Hospital were reviewed. All cases of third and fourth branchial remnants or pyriform sinus fistulae were identified. Clinical presentation, imaging, treatment, and outcome were recorded. Eight patients with a third or fourth branchial anomaly were identified and ranged in age from birth to 13 years. All anomalies were left sided. Presenting symptoms consisted of an asymptomatic cervical mass (n = 1), an infected mass (n = 5), neonatal respiratory distress (n = 1), and 1 incidental cyst found on magnetic resonance imaging. Ultrasonography was useful in suggesting the diagnosis in 7 cases. Barium swallow was performed in 3 patients with 2 positive results. Pharyngoscopy results showed the internal opening in 2 of 7 patients. A portion of the thyroid gland was resected in 6 patients. One patient has not yet undergone a definitive procedure. There was 1 recurrence in a patient whose pathology did not confirm a branchial remnant. The diagnosis and management of pyriform sinus anomalies are challenging. Ultrasound scan, computed tomography scan, barium swallow, and pharyngoscopy are all useful. The portion of thyroid involved in the fistula must be excised en bloc with the inflammatory mass, and the tract should be ligated at the level of the pharynx to minimize recurrence.


Epigenetic mechanisms elicited by nutrition in early life.
A growing number of studies focusing on the developmental origin of health and disease hypothesis have identified links among early nutrition, epigenetic processes and diseases also in later life. Different epigenetic mechanisms are elicited by dietary factors in early critical developmental ages that are able to affect the susceptibility to several diseases in adulthood. The studies here reviewed suggest that maternal and neonatal diet may have long-lasting effects in the development of non-communicable chronic adulthood diseases, in particular the components of the so-called metabolic syndrome, such as insulin resistance, type 2 diabetes, obesity, dyslipidaemia, hypertension, and CVD. Both maternal under- and over-nutrition may regulate the expression of genes involved in lipid and carbohydrate metabolism. Early postnatal nutrition may also represent a vital determinant of adult health by making an impact on the development and function of gut microbiota. An inadequate gut microbiota composition and function in early life seems to account for the deviant programming of later immunity and overall health status. In this regard probiotics, which have the potential to restore the intestinal microbiota balance, may be effective in preventing the development of chronic immune-mediated diseases. More recently, the epigenetic mechanisms elicited by probiotics through the production of SCFA are hypothesised to be the key to understand how they mediate their numerous health-promoting effects from the gut to the peripheral tissues.


Effects of long-term administration of Lactobacillus reuteri DSM-17938 on circulating levels of 5-HT and BDNF in adults with functional constipation.
Accumulated evidence shows that some probiotic strains ameliorate functional constipation (FC) via the modulation of specific gastrointestinal peptide pathways. The aims of this study were to investigate: (1) the effects of long-term administration of Lactobacillus reuteri (LR) DSM 17938 on the serum levels of serotonin (5-HT) and brain-derived neurotrophic factor (BDNF); (2) the possible link between 5-HT, BDNF, and specific constipation-related symptoms; (3) whether genetic variability at the 5-HTT gene-linked polymorphic region (5-HTTLPR) and BDNF Val66Met loci could be associated with serum 5-HT and BDNF variations. LR DSM 17938 was administered to 56 FC patients for 105 days in a randomised, double-blind manner. The fasting blood samples were collected during the randomisation visit (V1), at day 15 (induction period, V2), day 60 (intermediate evaluation, V3), and day 105 (V4) and the Constipaq questionnaire (the sum of Constipation Scoring System (CSS) and patient assessment constipation quality of life (PAC-QoL)) was administered. A group of healthy subjects was enrolled as controls (HC). At V1, the mean serum 5-HT level in the whole patient group was significantly higher (P=0.027) than in HC subjects, while serum BDNF did not. At the end of probiotic administration (V4), 5-HT and BDNF levels were significantly lower than the initial values (V1) (P=0.008 and P=0.015, respectively). 5-HT and BDNF serum concentration were significantly associated (r=0.355; P=0.007). Neither 5-HT nor BDNF serum levels correlated with the CSS item scores and with the PAC-QoL. Lastly, the regression analysis demonstrated that the presence of the S allele of the 5-HTTLPR accounted for the reduction in the 5-HT concentration at V4. In conclusion, the long-term administration of LR DSM 17938 demonstrated that such a probiotic strain could improve FC by affecting 5-HT and BDNF serum concentrations.


ANTIBODIES AGAINST INFLUENZA VIRUS TYPES A AND B IN CANADIAN SEALS.
Influenza viruses have been reported from marine mammals worldwide, particularly in pinnipeds, and have caused mass mortalities of seals in North America and Europe. Because influenza viruses in marine mammals can be zoonotic, our objective was to examine Canadian phocids for exposure to influenza A and B viruses in order to understand health risks to wild populations as well as to humans who consume or handle these animals. Blood was collected from 394 seals in eastern Canada from 1994 to 2005. Sera were screened for exposure to influenza viruses in three resident species of seals: harbour, Phoca vitulina (n=66); grey, Halichoerus grypus (n=82); ringed, Phoca hispida (n=2); and two migrant species: harp, Pagophilus groenlandica (n=206) and hooded, Cystophora cristata (n=38). Included were samples from captive grey (n=1) and harbour seals (n=8) at two aquaria. Sera were prescreened using indirect enzyme-linked immunosorbent assay (ELISA), and antibodies against influenza A virus were confirmed using a commercial competitive ELISA (IDEXX Europe B.V.). A subset of influenza A virus positive sera was used to determine common virus subtypes recognized by sera using reference strains. All positive sera in the indirect ELISA reacted with influenza A virus subtypes H3, H4, and H10 using a hemagglutination inhibition assay. Sera from harbour, grey, harp, and hooded seals had antibodies against influenza A and influenza B viruses (some cross-reactivity occurred). Overall, 33% (128/385) of wild seals were seropositive to influenza viruses, with the highest seroprevalence in harp (42%) followed by harbour (33%), grey (23%), and hooded (11%) seals. Antibodies were detected in both sexes and most age classes of wild seals. Two of eight captive harbour seals were seropositive to influenza B virus and four had cross-reactions to influenza A and B viruses. This study reports antibodies against influenza A and B viruses in four seal species from the same geographic area in eastern Canada.


Knowledge about the Scottish Mental Health Act in a general hospital setting.
Clinical experience suggests that non-psychiatrists' knowledge of mental health legislation in relation to emergency detention is inadequate. However, most non-psychiatrists will use this legislation at some point in their career. A questionnaire about emergency detention legislation was circulated to non-psychiatric medical staff to test their knowledge of the provisions relevant to the general hospital. Two hundred and seventy four medical staff replied (a response rate of 82%). Results showed that an understanding of detention procedures and applicability were markedly deficient, irrespective of grade, specialty or experience. The results indicate a need for increased education and awareness about emergency detention legislation especially in view of the implementation of the Mental Health (Scotland) Act (2003).


Adult dental epithelial stem cell-derived organoids deposit hydroxylapatite biomineral.
Ameloblasts are specialized cells derived from the dental epithelium that produce enamel, a hierarchically structured tissue comprised of highly elongated hydroxylapatite (OHAp) crystallites. The unique function of the epithelial cells synthesizing crystallites and assembling them in a mechanically robust structure is not fully elucidated yet, partly due to limitations with in vitro experimental models. Herein, we demonstrate the ability to generate mineralizing dental epithelial organoids (DEOs) from adult dental epithelial stem cells (aDESCs) isolated from mouse incisor tissues. DEOs expressed ameloblast markers, could be maintained for more than five months (11 passages) in vitro in media containing modulators of Wnt, Egf, Bmp, Fgf and Notch signaling pathways, and were amenable to cryostorage. When transplanted underneath murine kidney capsules, organoids produced OHAp crystallites similar in composition, size, and shape to mineralized dental tissues, including some enamel-like elongated crystals. DEOs are thus a powerful in vitro model to study mineralization process by dental epithelium, which can pave the way to understanding amelogenesis and developing regenerative therapy of enamel.


The role of dysfunctional attentional control networks in visual misperceptions in Parkinson's disease.
Visual misperceptions and hallucinations represent a problematic symptom of Parkinson's disease. The pathophysiological mechanisms underlying these symptoms remain poorly understood, however, a recent hypothesis has suggested that visual misperceptions and hallucinations may arise from disrupted processing across attentional networks. To test the specific predictions of this hypothesis, 22 patients with Parkinson's disease underwent 3T fMRI while performing the Bistable Percept Paradigm, a task that has previously been shown to identify patients with hallucinations. Subjects are required to study a battery of randomly assigned "monostable" and "bistable" monochromatic images for the presence or absence of a bistable percept. Those patients who scored a high percentage of misperceptions and missed images on the task were less able to activate frontal and parietal hubs of the putative Dorsal Attention Network. Furthermore, poor performance on the task was significantly correlated with the degree of decreased activation in a number of these hubs. At the group level, the difference between processing a bistable versus a monostable cue was associated with increased recruitment of the anterior insula. In addition, those patients with impaired performance on the paradigm displayed decreased resting state functional connectivity between hubs of the Ventral and Dorsal Attention Networks. These same patients had significantly decreased gray matter in the insula bilaterally. In addition, a combined analysis of the separate neuroimaging approaches revealed significant relationships across the impaired networks. These findings are consistent with specific predictions from a recently proposed hypothesis that implicates dysfunction within attentional networks in Parkinsonian hallucinations.


Continuous Estimation of Annual Committed Effective Dose of Radioactive Cesium by Market Basket Study in Japan from 2013 to 2019 after Fukushima Daiichi Nuclear Power Plant Accident.
Radionuclide contamination in foods has been a great concern after the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. To estimate time trends of daily intake and annual committed effective dose of radionuclides after the accident, radioactive cesium (r-Cs; 134Cs and 137Cs) and potassium-40 (40K) in market basket (MB) samples prepared at 6-month intervals in periods from September 2013 to March 2019 in 15 regions of Japan were analyzed using γ-ray spectrometry. The annual committed effective dose of r-Cs, calculated at non-detected radionuclide levels assumed to be half the limit of detection (LOD), appeared to decrease gradually in 11 regions close to the FDNPP that were more likely to be affected by the accident. Differences in doses among the 15 regions were large just after the accident, but gradually decreased. In particular, 134Cs has not been detected in any MB sample in any region since September 2018, and annual committed effective dose from 134Cs in all regions was mostly constant at around 0.3 μSv/year (given the respective LODs). The maximum annual committed effective dose of r-Cs in this study was decreased from 2.7 μSv/year in September 2013 to 1.0 μSv/year in March 2019. In contrast, the range of annual committed effective dose of 40K varied from approximately 150 to 200 μSv/year during that time frame and did not change much throughout the period of this study. Although annual committed effective doses of r-Cs in regions close to the FDNPP appeared to be higher than in regions far from the FDNPP, doses in all regions are remaining at a much lower levels than the intervention exemption level, 1 mSv/year, in foods in Japan.


A Precise Hip Protection System with Multi-scale Fall Warning Algorithm Based on Offset Displacement.
Falls occur frequently in daily life and the damage to the body is irreversible. Therefore, it is crucial to implement timely and effective warning and protection systems for falls to minimize the damage caused by falls. Currently, the fall warning algorithm has shortcomings such as low recognition rates for falls and fall-risk movements and insufficient lead-time, the time before the subject impacts the floor, making it difficult for falling protection devices to function effectively. In this study, a multi-scale falls warning algorithm based on offset displacement is built, and a hip protection system is designed. The performance of the algorithm and the system is validated using 150 falling and 500 fall-risk actions from 10 volunteers. The results showed that the recognition accuracy for falling actions is 98.7% and the recognition accuracy for fall-risk actions is 99.4%, with an average lead-time of 402ms. The protection rate for falling movements reached 98.7%. This proposed algorithm and hip protection system have the potential to be applied in elderly communities, hospitals, and homes to reduce the damage caused by falls.Clinical Relevance- This study provides important reference for clinicians in analyzing fall behaviors to patients at risk of falls in clinical settings, offering valuable technical support for ensuring the safety of patients in danger of falling. It also contributes to further promoting the development of falling-prevention medical devices.


The American 'Patterns of Care' Study: a model for the assessment of the quality of patient care in radiation oncology.
There are well documented manpower and equipment deficiencies in the specialty of Radiation Oncology in Australia. These deficiencies persist despite numerous reports over the last decade detailing their existence. The Patterns of Care Study (PCS) from the United States of America (USA) is a unique, comprehensive review of Radiation Oncology services in that country. It has clearly demonstrated that substantial benefits not only in survival but also in reduced treatment morbidity accrue when a full range of modern techniques and equipment is used. The present paper is a review of the PCS: this study has immense implications for patient care in particular and cancer services in general, in Australia. There seems little point in continuing to fund cancer research in the hope of gains in survival rates while continuing to ignore the fact that gains of a similar magnitude can be obtained by the maximum use of current knowledge and the provision of modern equipment.


Body composition, cardiometabolic risk factors, physical activity, and inflammatory markers in premenopausal women after a 10-year follow-up: a MONET study.
Menopausal transition and postmenopause are usually associated with changes in body composition and a decrease in physical activity energy expenditure (PAEE). This study investigated body composition, cardiometabolic risk factors, PAEE, and inflammatory markers in premenopausal women after a 10-year follow-up. In all, 102 premenopausal women participated in the 5-year observational longitudinal Montreal Ottawa New Emerging Team (MONET) study. This present substudy included 48 participants (age: 60.0 ± 1.7 years; body mass index: 23.2 ± 2.2 kg/m) 6.0 ± 0.3 years after completion of the initial MONET study. Measures included body composition, waist circumference (WC), fasting glucose and insulin levels, insulin sensitivity (QUICKI model), plasma lipid levels, PAEE, and inflammatory markers. Compared with baseline measures of the MONET study, analyses revealed no significant increase in body weight, although there were significant increases in WC, fat mass (FM), % FM, total cholesterol, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol, haptoglobin, apolipoprotein B, ferritin, adiponectin, and soluble cluster of differentiation 14 (all P < 0.001) after the 10-year follow-up. However, significant decreases were observed for fat-free mass, PAEE, fasting glucose levels, interleukin-8 levels, and soluble tumor necrosis factor receptors 1 and 2 (sTNFR-1 and sTNFR-2) levels (all P < 0.05). To determine the effect of postmenopausal years, data were restructured based on final menstrual period (FMP), and one-way analyses of variance were performed.Waist circumference, % FM, total cholesterol, high-density lipoprotein cholesterol, apolipoprotein B, ferritin, adiponectin, and soluble cluster of differentiation 14 were higher in early and late postmenopausal periods in these women. sTNFR-1 and sTNFR-2 levels were higher at the FMP and early postmenopausal years as compared with the late postmenopausal periods. Finally, interleukin-8 levels were lower in years after FMP. The number of years elapsed since the FMP can affect body composition, cardiometabolic risk factors, and inflammatory markers in healthy premenopausal women going through menopausal transition and postmenopausal periods.


Wind velocities on venus: vector determination by radio interferometry.
To determine the wind directions and speeds on Venus, as each Pioneer probe fell to the surface we tracked its motion in three dimensions using a combination of Doppler and long-baseline radio interferometric methods. Preliminary results from this tracking, coupled with results from test observations of other spacecraft, enable us to estimate the uncertainties of our eventual determinations of the velocity vectors of the probes with respect to Venus. For altitudes below about 65 kilometers and with time-averaging over 100-second intervals, all three components of the velocity should have errors of the order of 0.3 meter per second or less.


Starvation-induced MTMR13 and RAB21 activity regulates VAMP8 to promote autophagosome-lysosome fusion.
Autophagy, the process for recycling cytoplasm in the lysosome, depends on membrane trafficking. We previously identified Drosophila Sbf as a Rab21 guanine nucleotide exchange factor (GEF) that acts with Rab21 in endosomal trafficking. Here, we show that Sbf/MTMR13 and Rab21 have conserved functions required for starvation-induced autophagy. Depletion of Sbf/MTMR13 or Rab21 blocked endolysosomal trafficking of VAMP8, a SNARE required for autophagosome-lysosome fusion. We show that starvation induces Sbf/MTMR13 GEF and RAB21 activity, as well as their induced binding to VAMP8 (or closest Drosophila homolog, Vamp7). MTMR13 is required for RAB21 activation, VAMP8 interaction and VAMP8 endolysosomal trafficking, defining a novel GEF-Rab-effector pathway. These results identify starvation-responsive endosomal regulators and trafficking that tunes membrane demands with changing autophagy status.


Leveraging the Outpatient Pharmacy to Reduce Medication Waste in Pediatric Asthma Hospitalizations.
Previous local quality improvement focused on discharging patients with inhaled corticosteroids (ICS) "in-hand" decreased healthcare reutilization after hospitalization for an asthma exacerbation. However, as a result of these new processes, some patients admitted for an asthma exacerbation received more than one ICS inhaler during their admission, contributing to medication waste and potential patient confusion regarding their discharge medication regimen. We sought to decrease this waste. We conducted a quality improvement project to reduce the prescribing of multiple ICS inhalers to patients at a large academic children's hospital. Our primary outcome measure was the monthly percentage of patients admitted with an asthma exacerbation who were administered more than one ICS inhaler. A secondary outcome measure evaluated the reliability of the new process of using the hospital-based outpatient pharmacy to supply ICS "in-hand" and verify insurance coverage. After the process map review, we hypothesized a delay in the initial ICS treatment decision would allow for both a finalized discharge medication plan and a standardized process to verify outpatient insurance coverage. The mean percentage of patients receiving more than one ICS inhaler decreased from our baseline of 7.4% to 0.7%. Verification of outpatient prescription insurance coverage via the outpatient pharmacy increased from 0.7% to 50%. The average inpatient cost (average wholesale price) for ICS decreased by 62% to $90.25. Our process change to use the outpatient pharmacy to dispense and verify insurance coverage for ICS medication was associated with a reduction in medication waste during admission for an asthma exacerbation.


Life expectancy with and without cognitive impairment among Brazilian older adults.
Estimating the life expectancy with and without cognitive impairment in an older adult population is critical for understanding the burden of illness on individuals and their families, the health care system, and society at large. This paper presents and compares estimates of life expectancy with and without cognitive impairment for the noninstitutionalized population ages 60 years and older in São Paulo, Brazil, for the years 2000 and 2010. Life expectancy with and without cognitive impairment was calculated using the Sullivan method and prevalence estimates from data collection at two points (2000 and 2010) of the Health, Well-Being, and Aging (SABE) Study. Results indicate that 60-year-old men in São Paulo in 2000 could expect to live 14.8 years and women 17.9 years without cognitive impairment. By 2010, life expectancy without cognitive impairment had increased to 17.1 years for men and 20.0 years for women. Length of life with cognitive impairment differed by gender (2.3 years for men and 3.7 years for women at age 60 in 2010). However, the absolute number of years with cognitive impairment remained relatively constant with age. The results indicate a trend for improvements in life expectancy without cognitive impairment over time in São Paulo. Adults in Brazil still face many years of cognitive impairment in their older years, particularly when compared with estimates from developed countries.


TLR and B cell receptor signals to B cells differentially program primary and memory Th1 responses to Salmonella enterica.
Protective Th1 responses to Salmonella enterica do not develop in the absence of B cells. Using chimeric mice, we dissect the early (innate) and late (cognate) contributions of B cells to Th programming. B cell-intrinsic MyD88 signaling is required for primary effector Th1 development, whereas Ag-specific BCR-mediated Ag presentation is necessary for the development of memory Th1 populations. Programming of the primary T cell response is BCR/B cell MHC II independent, but requires MyD88-dependent secretion of cytokines by B cells. Chimeras in which B cells lack IFN-gamma or IL-6 genes make impaired Th1 or Th17 responses to Salmonella.


Alternative outcome measures in young total hip arthroplasty patients: a prospective cohort study.
In this prospective multicentre cohort study we studied subjects younger than 60 years of age scheduled for primary total hip arthroplasty (THA). The study assessed patients' overall satisfaction, fulfillment of preoperative expectations, the effect on socioeconomic parameters, and quality of sex-life. Questionnaires including Oxford Hip Score (OHS) and SF-36 were evaluated preoperatively and 3, 6 and 12 months postoperatively. OHS and SF-36 showed significant improvements (p<0.028, Wilcoxon Signed Rank test). THA did not affect the patients' socioeconomic status. Increased frequency of intercourse or better abilities in intercourse positions were experienced by 18 of 39 females due to reduced pain and increased range of motion. Patients sexually active before THA surgery remained active. These findings constitute important new information to young patients and surgeons during the decision making process.


Structural basis of an engineered dual-specific antibody: conformational diversity leads to a hypervariable loop metal-binding site.
To explore dual-specificity in a small protein interface, we previously generated a 'metal switch' anti-RNase A VHH antibody using a combinatorial histidine library approach. While most metal-binding sites in proteins are found within rigid secondary structure, the engineered VHH antibody (VHH(metal)), which contained three new histidine residues, possessed metal-binding residues within the flexible hypervariable loops. Here, crystal structure analysis of the free and bound states of VHH(metal) reveals the structural determinants leading to dual-function. Most notably, CDR1 is observed in two distinct conformations when adopting the metal or RNase A bound states. Furthermore, mutagenesis studies revealed that one of the engineered residues, not located in the metal-binding pocket, contributed indirectly to metal recognition, likely through influencing CDR1 conformation. Despite these changes, VHH(metal) possesses a relatively minor energetic penalty toward binding the original antigen, RNase A (~1 kcal/mol), where the engineered gain-of-function metal-binding residues are observed to possess a mix of favorable and unfavorable contributions towards RNase A recognition. Ultimately, the conformationally distinct metal-switch interface architecture reflects the robust, library-based strategy used to produce VHH(metal). These results also suggest that even small protein interfaces, such as VHH, may be structurally and energetically forgiving in adopting novel function, while maintaining original function.


Future pharmacological treatment of cystic fibrosis.
Cystic fibrosis (CF) is an autosomal recessive disorder that is caused by over 850 different mutations in the CF gene. It is useful to group these mutations according to the defect that results in the CFTR mRNA or protein. New pharmacological treatments targeted towards specific mutations that are relatively common are being developed. Class I mutations do not produce CFTR protein because of a premature stop signal in the CFTR DNA. These null mutations can be corrected by certain aminoglycosides which cause the aberrant stop signal to be skipped. Mutations leading to a CFTR protein that attains an unstable structure shortly after translation in the endoplasmic reticulum form class II. Class II mutations can be restored to the protein trafficking pathway by manipulation of chaperone protein/CFTR interactions with chemical chaperones or drugs that affect gene regulation such as the butyrates. Production of a CFTR with reduced Cl(-) transport on the basis of abnormal regulation of the chloride channel is the basis of class III. Genistein can overcome this block in regulation. Mutations that partially reduce chloride conductance through CFTR (class IV) can be stimulated with milrinone, which is a phosphodiesterase inhibitor. Finally, mutations that lead to a severe reduction in normal CFTR protein form class V. Increased levels of CFTR could be generated with the butyrates or supplemented with gene therapy. Although most of the reported mutations in CFTR are rare and unclassified, it may be possible to use genotype-phenotype correlations to determine the best approach.


Characteristics associated with genetic counseling referral and BRCA1/2 testing among women in a large integrated health system.
Evidence shows underutilization of cancer genetics services. To explore the reasons behind this underutilization, this study evaluated characteristics of women who were referred for genetic counseling and/or had undergone BRCA1/2 testing. An ovarian cancer risk perception study stratified 16,720 eligible women from the Henry Ford Health System into average-, elevated-, and high-risk groups based on family history. We randomly selected 3,307 subjects and interviewed 2,524 of them (76.3% response rate). Among the average-, elevated-, and high-risk groups, 2.3, 10.1, and 20.2%, respectively, reported genetic counseling referrals, and 0.8, 3.3, and 9.5%, respectively, reported having undergone BRCA testing. Personal breast cancer history, high risk, and perceived ovarian cancer risk were associated with both referral and testing. Discussion of family history with a doctor predicted counseling referral, whereas belief that family history influenced risk was the strongest BRCA testing predictor. Women perceiving their cancer risk as much higher than other women their age were twice as likely (95% confidence interval: 2.0-9.6) to report genetic counseling referral. In a health system with ready access to cancer genetic counseling and BRCA testing, women who were at high risk underutilized these services. There were strong associations between perceived ovarian cancer risk and genetic counseling referral, and between a belief that family history influenced risk and BRCA testing.


Description of the Southern swamp leech Philobdella floridana (Verrill, 1874) (Annelida: Hirudinea) from the Outer Banks, North Carolina, USA, with a revision of the species.
The jawed leech Philobdella floridana (Verrill, 1874) occurs widely in swamps of the southeastern United States. The discovery of a population of P. floridana in Lake Phelps, an isolated lake in the Albemarle Peninsula in the Outer Banks region of North Carolina, is by far the northernmost record for this species. Description of the Lake Phelps leech in this paper is the first comprehensive account of the internal anatomy of P. floridana. Comparable internal descriptions of P. floridana from other geographic regions are also presented for the first time. This new understanding of the internal features of this species is the basis of a taxonomic revision of P. floridana. All specimens examined in this study possessed remarkably similar jaws and teeth within the range of 22-26 per jaw, regardless of the size or origin of the leech. All mature specimens possessed external copulatory slits and pores which uniquely characterise the genus Philobdella. This study shows for the first time that these external features are attributable to a complex internal network of gastropores emanating from the crop caeca of the genital segments. Nonetheless, the reproductive anatomy of this species is by far the most rudimentary of any Hirudinidae in North America. A major finding of this study is that the Lake Phelps population differs significantly from other known P. floridana in one significant respect. Specifically, the gut has an intimate anatomical association with the nephridia, an enteronephric feature previously unknown within the Hirudinea. At least in this study, enteronephry may be confined to Lake Phelps, leaving open the possibility that this population of P. floridana may be endemic to this relict lake which has a history of endemicity.


NF1 patient missense variants predict a role for ATM in modifying neurofibroma initiation.
In Neurofibromatosis type 1, NF1 gene mutations in Schwann cells (SC) drive benign plexiform neurofibroma (PNF), and no additional SC changes explain patient-to-patient variability in tumor number. Evidence from twin studies suggests that variable expressivity might be caused by unidentified modifier genes. Whole exome sequencing of SC and fibroblast DNA from the same resected PNFs confirmed biallelic SC NF1 mutations; non-NF1 somatic SC variants were variable and present at low read number. We identified frequent germline variants as possible neurofibroma modifier genes. Genes harboring variants were validated in two additional cohorts of NF1 patients and by variant burden test. Genes including CUBN, CELSR2, COL14A1, ATR and ATM also showed decreased gene expression in some neurofibromas. ATM-relevant DNA repair defects were also present in a subset of neurofibromas with ATM variants, and in some neurofibroma SC. Heterozygous ATM G2023R or homozygous S707P variants reduced ATM protein expression in heterologous cells. In mice, genetic Atm heterozygosity promoted Schwann cell precursor self-renewal and increased tumor formation in vivo, suggesting that ATM variants contribute to neurofibroma initiation. We identify germline variants, rare in the general population, overrepresented in NF1 patients with neurofibromas. ATM and other identified genes are candidate modifiers of PNF pathogenesis.


Effect of clofibrate on cholesterol and lipid metabolism in ethanol - treated mice.
The effects of chronic clofibrate administration were studied in long-term ethanol-fed an control mice. Long-term administration of ethanol to adult-mice resulted in an increase in liver and plasma total triglycerides and hepatic cholesterol levels as compared to the corresponding controls. Cloribrate feeding resulted in about 15-25% decrease in hepatic and plasma cholesterol levels in the ethanol-fed group. The administration of ethanol led to a rise in levels of hepatic triglycerides, plasma triglycerides, plasma free fatty acids and hepatic alpha-glycerophosphate. Treatment with clofibrate partially prevented the changes in the concentration of these metabolites and resulted in a significant increase in the activity of hepatic alpha-glycerophosphate dehydrogenase. Clofibrate administration resulted in about 20% increase in the rate of 14-C-cholesterol oxidation to 14-CO2. It is concluded that while clofibrate is able to retard ethanol-induced lipid accumulation in the liver, it is unable to completely prevent hepatic lipid accumulation.


[The lobe of Zuckerkandl: an important sign of recurrent laryngeal nerve].
The accurate acquaintance of the anatomy of the thyroid gland allows reduction of complications to interventions of thyroidectomy, where for the existing topography, the nervous and vascular structures could result vulnerable. The identification of Zuckerkandl's tuberculum could reduce the lesions to the recurrent laryngeal nerve, for constant relationship between the recurrent laryngeal nerve and tuberculum. The Authors have studied 605 patients underwent to thyroid surgery for benign or malignant thyroid diseases. The aim was to verify if the incidence of recurrent nerve lesions can be reduced with the identification of the Zuckerkandl's tuberculum and with a knowledge of its anatomical relationships with vascular, nervous and glandular structures near the thyroid. The Zuckerkandls tuberculum was found in the majority of the cases, with prevalence to the right. Its identification has allowed an immediate and safe identification of recurrent laryngeal nerve, with setting of time of the operation and especially with setting of possible injury to the recurrent nerves. The lobe of Zuckerkandl is the extension of the lateral lobes of the thyroid, composed of thyroid tissue only and so it can be interested in thyroid lesions. This tubercle is considered a constant anatomical landmark for the recurrent laryngeal nerve and the superior parathyroid glands. The knowledge of the lobe of Zuckerkandl is essential to perform "safety thyroidectomy", without injury for the vascular and nervous structures. TZ identification is not always easy and/or possible but, when that happens become aware of possible, systematically, the isolation of the recurrent nerve and of the superior parathyroid gland, preserve such structures from possible lesions in surgery of the thyroid gland.


A Systematic Approach to Bacterial Phylogeny Using Order Level Sampling and Identification of HGT Using Network Science.
Reconstructing and visualizing phylogenetic relationships among living organisms is a fundamental challenge because not all organisms share the same genes. As a result, the first phylogenetic visualizations employed a single gene, e.g., rRNA genes, sufficiently conserved to be present in all organisms but divergent enough to provide discrimination between groups. As more genome data became available, researchers began concatenating different combinations of genes or proteins to construct phylogenetic trees believed to be more robust because they incorporated more information. However, the genes or proteins chosen were based on ad hoc approaches. The large number of complete genome sequences available today allows the use of whole genomes to analyze relationships among organisms rather than using an ad hoc set of genes. We present a systematic approach for constructing a phylogenetic tree based on simultaneously clustering the complete proteomes of 360 bacterial species. From the homologous clusters, we identify 49 protein sequences shared by 99% of the organisms to build a tree. Of the 49 sequences, 47 have homologous sequences in both archaea and eukarya. The clusters are also used to create a network from which bacterial species with horizontally-transferred genes from other phyla are identified.


Statistical estimates of absenteeism attributable to seasonal and pandemic influenza from the Canadian Labour Force Survey.
As many respiratory viruses are responsible for influenza like symptoms, accurate measures of the disease burden are not available and estimates are generally based on statistical methods. The objective of this study was to estimate absenteeism rates and hours lost due to seasonal influenza and compare these estimates with estimates of absenteeism attributable to the two H1N1 pandemic waves that occurred in 2009. Key absenteeism variables were extracted from Statistics Canada's monthly labour force survey (LFS). Absenteeism and the proportion of hours lost due to own illness or disability were modelled as a function of trend, seasonality and proxy variables for influenza activity from 1998 to 2009. Hours lost due to the H1N1/09 pandemic strain were elevated compared to seasonal influenza, accounting for a loss of 0.2% of potential hours worked annually. In comparison, an estimated 0.08% of hours worked annually were lost due to seasonal influenza illnesses. Absenteeism rates due to influenza were estimated at 12% per year for seasonal influenza over the 1997/98 to 2008/09 seasons, and 13% for the two H1N1/09 pandemic waves. Employees who took time off due to a seasonal influenza infection took an average of 14 hours off. For the pandemic strain, the average absence was 25 hours. This study confirms that absenteeism due to seasonal influenza has typically ranged from 5% to 20%, with higher rates associated with multiple circulating strains. Absenteeism rates for the 2009 pandemic were similar to those occurring for seasonal influenza. Employees took more time off due to the pandemic strain than was typical for seasonal influenza.


Mutation of the arginine finger in the active site of Escherichia coli DbpA abolishes ATPase and helicase activity and confers a dominant slow growth phenotype.
Escherichia coli DEAD-box protein A (DbpA) is an ATP-dependent RNA helicase with specificity for 23S ribosomal RNA. Although DbpA has been extensively characterized biochemically, its biological function remains unknown. Previous work has shown that a DbpA deletion strain is viable with little or no effect on growth rate. In attempt to elucidate a phenotype for DbpA, point mutations were made at eleven conserved residues in the ATPase active site, which have exhibited dominant-negative phenotypes in other DExD/H proteins. Biochemical analysis of these DbpA mutants shows the expected decrease in RNA-dependent ATPase activity and helix unwinding activity. Only the least biochemically active mutation, R331A, produces small colony phenotype and a reduced growth rate. This dominant slow growth mutant will be valuable to determine the cellular function of DbpA.


Reaching Adolescents for Prevention: The Role of Pediatric Emergency Department Health Promotion Advocates.
Almost 200,000 adolescents visit US emergency departments (EDs) yearly for conditions involving underage drinking but receive no follow-up referral. Other health risk behaviors resulting in sexually transmitted infections, car crashes, and assault-related injury are common among adolescents. A pediatric ED (PED) visit presents an opportunity to discuss and promote prevention. We report here on implementation of a new PED navigator/extender role, the Health Promotion Advocate (HPA). Health Promotion Advocates surveyed patients to identify health risks, stresses, and needs. A positive screen triggered a brief conversation containing the following elements: permission to discuss risks/needs; exploration of context (a typical day in your life); brief feedback (information and norms); exploration of benefits and consequences of risk behaviors; assessment of readiness to change; calling up assets, instilling hope; discussing challenges of change; negotiating a menu of options and prescription for change; referrals to primary care, community resources; and treatment services as indicated. During 2009-2013, HPAs screened 2149 PED patients aged 14 to 21 years and referred 834 for an array of services (eg, primary care, mental health, insurance, personal safety, human immunodeficiency virus testing, general education diploma (GED), employment, housing, and food pantries) to address reported health risks; 785 screened positive for at-risk substance use (53% female, 36% without primary care). Among them, 636 received a brief intervention; 546 were referred to specialized substance abuse treatment. Two case studies are presented to illustrate the engagement and referral process. Health Promotion Advocates working as PED team members can extend PED services beyond the scope of the presenting complaint.


Thermal Boundary Characteristics of Homo-/Heterogeneous Interfaces.
The interface of two solids in contact introduces a thermal boundary resistance (TBR), which is challenging to measure from experiments. Besides, if the interface is reactive, it can form an intermediate recrystallized or amorphous region, and extra influencing phenomena are introduced. Reactive force field Molecular Dynamics (ReaxFF MD) is used to study these interfacial phenomena at the (non-)reactive interface. The non-reactive interfaces are compared using a phenomenological theory (PT), predicting the temperature discontinuity at the interface. By connecting ReaxFF MD and PT we confirm a continuous temperature profile for the homogeneous non-reactive interface and a temperature jump in case of the heterogeneous non-reactive interface. ReaxFF MD is further used to understand the effect of chemical activity of two solids in contact. The selected Si/SiO 2 materials showed that the TBR of the reacted interface is two times larger than the non-reactive, going from 1 . 65 × 10 - 9 to 3 . 38 × 10 - 9 m 2 K/W. This is linked to the formation of an intermediate amorphous layer induced by heating, which remains stable when the system is cooled again. This provides the possibility to design multi-layered structures with a desired TBR.


Risk Assessment of Repetitive Movements in Olive Growing: Analysis of Annual Exposure Level Assessment Models with the OCRA Checklist.
In Italy, one of the main agricultural crops is represented by the cultivation of olive trees. Olive cultivation characterizes the Italian agricultural landscape and national agricultural economics. Italy is the world's second largest producer of olive oil. Because olive cultivation requires the largest labor force in southern Italy, the aim of this research was to assess the risk of biomechanical overload of the workers' upper limbs. The objective, therefore, was to determine the level of risk that workers are exposed to in each phase of the production process. In Calabria, the second most important region in Italy for both the production of olive oil and cultivated area, there are 113,907 olive farms (83% of all farms) and 250,000 workers. To evaluate the risk of repetitive movements, all of the work tasks performed by workers on 100 farms in Calabria were analyzed. A total of 430 workers were interviewed over the four-year research period. To evaluate the level of exposure to repetitive movements, the OCRA (occupational repetitive actions) checklist was adopted. This checklist was the primary analytical tool during the preliminary risk assessment and in a given working situation. The analysis suggested by the OCRA checklist starts with pre-assigned scores (increasing in value with intensification of risk) for each of four main risk factors and additional factors. Between 2010 and 2013, surveys were conducted using the OCRA checklist with the aim of verifying musculoskeletal risks. The results obtained from the study of 430 workers allowed us to identify the level of exposure to risk. This analysis was conducted in the workplace to examine in detail the repetitive movements performed by the workers. The research was divided into two phases: first to provide preliminary information on the different tasks performed in olive growing, and second to assign a percentage to each task of the total hours worked in a year. Based on the results, this method could well become a tool for implementing prevention measures for all workers involved in farming, not just those in olive production. Exposure to work-related musculoskeletal risks (extreme posture, repetition, high muscle loads) appears to be significant for workers in olive production. In the cultivation of olive trees, many tasks require prolonged and intense physical efforts that can increase risk. The most problematic work tasks were pruning and harvesting, while fertilization showed the lowest risk. The survey showed the need for a suitable study of olive growing to adapt prevention measures to specific aspects of agriculture as compared to other employment sectors.


Heart rate variability analysis in sheep affected by transmissible spongiform encephalopathies.
The function of the autonomic nervous system can be assessed by determining heart rate variability (HRV), which is impaired in some brainstem diseases in humans. Transmissible spongiform encephalopathies (TSEs) in sheep are diseases characterised by accumulation of disease-associated prion protein in the brainstem, including nuclei of the parasympathetic nervous system. This study was undertaken to assess whether analysis of HRV can be used as an aid in the diagnosis of TSEs in clinically affected, naturally or experimentally infected sheep. When HRV indices were compared between 41 clinical TSE cases (18 sheep infected with scrapie and 23 sheep infected with bovine spongiform encephalopathy), 11 control sheep and six sheep reported as scrapie suspects or dosed with BSE brain homogenate, which were not confirmed as TSE cases by postmortem tests, no significant differences were found between the groups. Median heart rate was significantly different but only when sheep were grouped by gender: it was higher in female TSE cases than in control sheep and higher in female than castrated male ovine classical BSE cases. HRV analysis was not useful as a diagnostic aid for TSEs of sheep.


Nitric oxide synthase inhibition attenuates cutaneous vasodilation during postmenopausal hot flash episodes.
The purpose of this study was to test the hypothesis that local inhibition of nitric oxide and prostaglandin synthesis attenuates cutaneous vasodilator responses during postmenopausal hot flash episodes. Four microdialysis membranes were inserted into the forearm skin (dorsal surface) of eight postmenopausal women (mean +/- SD age, 51 +/- 7 y). Ringer solution (control), 10 mM ketorolac (Keto) to inhibit prostaglandin synthesis, 10 mM N-l-arginine methyl ester (l-NAME) to inhibit nitric oxide synthase, and a combination of 10 mM Keto + 10 mM l-NAME were each infused at the separate sites. Skin blood flow at each site was indexed using laser-Doppler flowmetry. Cutaneous vascular conductance (CVC) was calculated as laser-Doppler flux/mean arterial blood pressure and was expressed as a percentage of the maximal calculated CVC (CVCmax) obtained after infusion of 50 mM sodium nitroprusside at all sites at the end of the study. Data from 13 hot flash episodes were analyzed. At the control site, the mean +/- SD peak increase in CVC was 15.5% +/- 6% CVCmax units. This value was not different relative to the peak increase in CVC at the Keto site (13.0% +/- 5% CVCmax units; P = 0.09). However, the peak increase in CVC during hot flash episodes were attenuated at the l-NAME and l-NAME + Keto sites (7.4% +/- 4% and 8.7% +/- 7% CVCmax units, respectively) relative to both the control and the Keto sites (P < 0.05 for both comparisons). There were no significant differences in the peak increases in sweat rate between any of the sites (P = 0.24). These data demonstrate that the mechanism for cutaneous vasodilation during hot flash episodes has a nitric oxide component. Increases in CVC despite the inhibition of prostaglandin synthesis suggest that prostaglandins do not contribute to cutaneous vasodilation during hot flash episodes.


A microfluidic mixer with grooves placed on the top and bottom of the channel.
A new microfluidic mixer is presented consisting of a rectangular channel with grooves placed in the top and bottom. This not only increases the driving force behind the lateral flow, but allows for the formation of advection patterns that cannot be created with structures on the bottom alone. Chevrons, pointing in opposite directions on the top and bottom, are used to create a pair of vortices positioned side by side. Stripes running the width of the channel generate a pair of vertically stacked vortices. Computational fluid dynamics (CFD) simulations are used to model the behavior of the systems and provide velocity maps at cross-sections within the mixer. Experiments demonstrate the mixing that results when two segregated species enter the mixer side-by-side and pass through two cycles of the mixer (i.e., two alternating sets of four stripes and four chevrons).


Purchasing health care: an opportunity for a public-private partnership.
By recommending that the federal government take a leadership role in driving quality improvement in the U.S. health care system, the Institute of Medicine has taken a bold step, one that signals a significant policy shift. Through regulations and payments, government incentives have a profound effect on the practices of hospitals, clinicians, and other institutions. Private-sector employers sponsor coverage for 165 million people, and they will want a substantial voice in government strategies and actions. This paper recommends that the two sectors formally develop a strategic partnership, mediated through two powerful advisory boards, one at the level of the HHS secretary and the other at the level of the CMS administrator.


Overexpression of EPH receptor B2 in malignant mesothelioma correlates with oncogenic behavior.
Malignant pleural mesothelioma (MM) is an aggressive asbestos-associated malignancy with limited therapeutic options. This study describes the overexpression of Ephrin B2 receptor (EPHB2) in MM cell lines and tumors, and the effect of its manipulation on proliferative and invasive qualities of the disease. Using expression arrays, we investigated EPHB2 in MM tumors compared with normal mesothelium. EPHB2 and downstream target expression were evaluated using reverse-transcriptase polymerase chain reaction and immunoblotting methods. The biological significance of EPHB2 in MM was evaluated using in vitro functional assays with and without targeting by EPHB2-short hairpin RNA or blocking peptide in two mesothelioma cell lines, HP-1 and H2595. EPHB2 is overexpressed in all MM cell lines, but not in benign mesothelial cells, and is significantly elevated in MM tumor tissue compared with matched normal peritoneum. Targeted knockdown of EPHB2 in HP-1 and H2595 cell lines reduced its expression and that of EPHB2 downstream targets such as matrix metalloproteinase (MMP-2) and vascular endothelial growth factor, whereas caspase 2 and caspase 8 had increased expression. Inhibition of EPHB2 resulted in a significant decrease in scratch closure (1.25-fold-1.8-fold), proliferation (1.5-fold), and invasion (1.7-fold-1.8-fold) compared with the controls. Most notably, however, EPHB2 silencing resulted in a significant increase in apoptotic proteins and activity. EPHB2 seems to play an important role in MM pathogenesis and these findings indicate that EPHB2 could serve as a potential novel therapeutic target for treatment of the disease.


Effectiveness and limitations of local structural entropy optimization in the thermal stabilization of mesophilic and thermophilic adenylate kinases.
Local structural entropy (LSE) is a descriptor for the extent of conformational heterogeneity in short protein sequences that is computed from structural information derived from the Protein Data Bank. Reducing the LSE of a protein sequence by introducing amino acid mutations can result in fewer conformational states and thus a more stable structure, indicating that LSE optimization can be used as a protein stabilization method. Here, we describe a series of LSE optimization experiments designed to stabilize mesophilic and thermophilic adenylate kinases (AKs) and report crystal structures of LSE-optimized AK variants. In the mesophilic AK, thermal stabilization by LSE reduction was effective but limited. Structural analyses of the LSE-optimized mesophilic AK variants revealed a strong correlation between LSE and the apolar buried surface area. Additional mutations designed to introduce noncovalent interactions between distant regions of the polypeptide resulted in further stabilization. Unexpectedly, optimizing the LSE of the thermophilic AK resulted in a decrease in thermal stability. This destabilization was reduced when charged residues were excluded from the possible substitutions during LSE optimization. These observations suggest that stabilization by LSE reduction may result from the optimization of local hydrophobic contacts. The limitations of this process are likely due to ignorance of other interactions that bridge distant regions in a given amino acid sequence. Our results illustrate the effectiveness and limitations of LSE optimization as a protein stabilization strategy and highlight the importance and complementarity of local conformational stability and global interactions in protein thermal stability.


[Treatment of primary central nervous system lymphoma: recent advances].
Primary central nervous system lymphoma is a quite infrequent disease within lymphoproliferative disorders. Moreover, it must be taken into account that, even considering the progressive spreading of ameliorated diagnostic tools and skills, histology remains the hinge of a correct diagnosis (stereotactic cerebral biopsy). Hence, randomised clinical protocols with a sufficiently large number of patients enrolled are difficult to perform and a generalized consensus on the best therapeutic options is hard to achieve. In this article, the more frequent practical and clinical aspects regarding diagnosis and staging of this rare disease are discussed. Furthermore, we analyze the different therapeutic options that were considered in a relatively recent past in which the median survival from diagnosis was not longer than a year, working down the pathway to the present and, finally, to the near future, always considering both first line and salvage therapy.The goal remains to optimize and standardize the therapeutic approach in order to achieve a generalized consensus that should lead to better overall clinical results.


[Coronary artery reoperation].
Coronary artery reoperation has been performed in 51 patients (4%) including 5 second reoperations among 1,245 CABGs during past 9 years. There were 40 males and 11 females with a mean age of 58 year old ranging from 19 to 75 year old. Extent of coronary artery disease were; single vessel disease in 2 patients, double vessel disease in 18 patients, triple vessel disease in 22 patients and left main disease in 9 patients. Preoperative ejection fraction ranged from 0.24 to 0.69 (mean 0.48) and 21 patients (41%) showed unstable angina. Interval between primary CABG and reoperation ranged from 1 month to 20 years with a mean of 6 years. There were two groups which had early reoperation at the mean of 6 months (14 patients) and late reoperation at the mean of 9 years (37 patients) after primary CABG. Causes of reoperation varied from graft failure (29 patients), progression of native CAD (2 patients), both of them (19 patients), and incomplete revascularization (1 patient). There were 32 patients who had patent old graft at the time of reoperation. Re-entry approaches used were midsternal in 41 patients and left thoracotomy in 10 patients. The ITA, GEA, IEA and SV grafts were used in 30, 38, 5 and 28 patients, respectively. There were 4 patients who underwent CABG without cardiopulmonary bypass. The mean number of bypass was 2.1 and the mean duration of aortic crossclamp and cardiopulmonary bypass were 63 minutes and 114 minutes, respectively. As a result, there were 4 early and 1 late death. Postoperative angiography revealed that patency rate of ITA, GEA, IEA and SV grafts were 100% (22/22), 97% (30/31), 75% (3/4) and 87% (20/23), respectively. In conclusion, risk of coronary artery reoperation is still high and use of arterial graft is important to obtain high patency rate and low reoperation rate.


Factors related to follicular oxidative stress in intracytoplasmic sperm injection cycles and its effects on granulosa cells.
The aim of the present study was to investigate several common conditions that may potentially be correlated with follicular oxidative status during an intracytoplasmic sperm injection (ICSI) cycle and that include the serum oestrogen level on the day of oocyte pick-up, maternal age and pregnancy outcome. Patients that were enrolled in the study were classified randomly into three groups using their numerical order. The first group were classified based on maternal age (<35 and ≥35 years) (n = 398), the second group on the serum oestradiol (E2) level on the day of human chorionic gonadotropin (hCG) administration (levels >90th percentile and ≤ 90th percentile) (n = 491) and the third group on pregnancy outcome (positive/negative) (n = 376). The groups were matched for the other variables (stimulation protocol, dose of gonadotropin, duration of stimulation, antral follicle count, body mass index, basal follicle stimulating hormone (FSH), and E2 levels and day of hCG trigger) to prevent the possible contribution of those parameters to the results. Each group was matched for other variables (stimulation protocol, dose of gonadotrophin, duration of stimulation, antral follicle count, body mass index, basal FSH and E2 levels and day of hCG trigger) that may have affected the outcome, except for the parameter under investigation. Maternal age (P = 0.044,168 r = 0.418), oestrogen level on day of hCG administration (P = 0.001, r = 0.436) and pregnancy outcome (AUC = 0.65, P = 0.071) were found to be correlated with follicular oxidative status. The results obtained will help us to shield patients from possible situations that may cause oxidative stress and therefore adverse outcomes of an ICSI cycle.