Intravenous synthetic peptide MBP8298 delayed disease progression in an HLA Class II-defined cohort of patients with progressive multiple sclerosis: results of a 24-month double-blind placebo-controlled clinical trial and 5 years of follow-up treatment.
MBP8298 is a synthetic peptide with a sequence corresponding to amino acid residues 82-98 of human myelin basic protein (DENPVVHFFKNIVTPRT). It represents the immunodominant target for both B cells and T cells in multiple sclerosis (MS) patients with HLA haplotype DR2. Its administration in accordance with the principle of high dose tolerance results in long-term suppression of anti-myelin basic protein (MBP) autoantibody levels in the cerebrospinal fluid (CSF) of a large fraction of progressive MS patients. MBP8298 was evaluated in a 24-month placebo-controlled double-blinded Phase II clinical trial in 32 patients with progressive MS. The objective was to assess the clinical efficacy of 500 mg of MBP8298 administered intravenously every 6 months, as measured by changes in Expanded Disability Status Scale (EDSS) scores. Contingency analysis for all patients at 24 months showed no significant difference between MBP8298 and placebo-treatments (n = 32, P = 0.29). Contingency analysis in an HLA Class II defined subgroup showed a statistically significant benefit of MBP8298 treatment compared with placebo in patients with HLA haplotypes DR2 and/or DR4 (n = 20, P = 0.01). Long-term follow-up treatment and assessment of patients in this responder group showed a median time to progression of 78 months for MBP8298 treated patients compared with 18 months for placebo-treatment (Kaplan-Meier analysis, P = 0.004; relative rate of progression = 0.23). Anti-MBP autoantibody levels in the CSF of most MBP8298 treated patients were suppressed, but antibody suppression was not predictive of clinical benefit. Anti-MBP autoantibodies that reappeared in the CSF of one patient at 36 months, whilst under treatment with MBP8298, were not reactive with the MBP8298 peptide in vitro. The identification of a responder subgroup (62.5% of the patients in this study) enables a more efficient design of a large confirmatory clinical trial of MBP8298. The probability that patients with other less common HLA-DR haplotypes will respond to this treatment should not be ignored.


A National Analysis of Treatment Patterns and Outcomes for Patients 80 Years or Older With Esophageal Cancer.
The purpose of this study was to evaluate practice patterns and outcomes for patients 80 years or older with esophageal cancer using a nationwide cancer data base. Practice patterns for patients 80 years or older with stage I-IV esophageal cancer in the National Cancer Data Base from 2004 to 2014 were analyzed. Overall survival associated with different treatment strategies were evaluated using the Kaplan-Meier method and multivariable Cox proportional hazard models. In the study period, 40.5% and 46.2% of patients with stage I adenocarcinoma and squamous cell carcinoma, respectively, did not receive any treatment at all. Less than 11% (196/1,865) of patients with stage I-II disease underwent esophagectomy, even though surgery was associated with a better 5-year survival compared to no treatment (stage I: 47.3% [95% confidence interval [CI] 36.2-57.6%] vs 14.9% [95% CI: 11.2-19.1%]; stage II: 29.3% [95% CI 20.1-39.1%] vs 1.2% [95% CI: 0.1-5.5%]). Of the 1,596 (37.7%) patients with stage III disease who received curative-intent treatment (surgery or chemoradiation), the 5-year survival was significantly better than that of patients who received no treatment (11.9% [95% CI: 9.7-14.4% vs 4.3% [95% CI: 1.9-8.3%]). In this national analysis of patients 80 years and older with esophageal cancer, over 40% of patients with stage I disease did not receive treatment. Patients with stage I-III disease had better survival and risks and benefits of treatment for elderly patients should be discussed in a multidisciplinary setting.


Intestinal epithelial cells and T cells differentially recognize and respond to Candida albicans yeast and hypha.
Inflammatory bowel diseases (IBD) are a multifactorial disorder. Our understanding of the role of bacteria in the pathogenesis of IBD has increased substantially; however, only scarce data exist regarding the role of commensal fungi in maintaining intestinal homeostasis and triggering IBD. Candida albicans (C. albicans) is a member of the intestinal mycobiome and proposed to contribute to IBD pathogenesis. We aimed to investigate the influence of the two morphologies of C. albicans, yeast and hypha, on epithelial cells and T cells from IBD patients versus healthy controls. We found that C. albicans was recognized by both epithelial cells lines and T cells. In the intestinal epithelial cell line, Caco-2, response to hypha was different than to yeast cells, and this was mimicked by synthetic β-glucans and Pam3CSK4. Unstimulated T cells exhibited increased activation and pro-inflammatory cytokine secretion upon exposure, while there was no effect on apoptosis or proliferation. In contrast, C. albicans-challenged CD3-stimulated T-cells exhibited decreased activation, cytokine secretion, apoptosis, and proliferation, suggesting reciprocal responsiveness to C. albicans. Glycans alone did not mimic abovementioned influences on T cells, suggesting alternative modes of recognition. In conclusion, we provide evidence for glycan dependent and independent recognition of C. albicans by epithelial cells and T cells.


Prevalence of dementia and factors associated with dementia in rural Bangladesh: data from a cross-sectional, population-based study.
There are currently no published reports of dementia prevalence or factors associated with dementia occurrence in Bangladesh. The aims are to report the prevalence of definite and questionable dementia in rural Bangladesh, and examine factors potentially associated with dementia occurrence, including sociodemographic, clinical, social, and nutritional factors. We used data from a population-based, cross-sectional study from Matlab, in rural Bangladesh, on 471 persons aged 60+ years. Participants underwent a clinical examination including diagnosis of somatic disorders, and a structured interview including questions about sociodemographic and social factors. Nutritional status was measured with the Mini Nutritional Assessment, and blood tests were conducted to assess a range of nutritional and clinical aspects. Age- and sex-specific dementia prevalence was calculated. Crude and adjusted logistic regression was used to examine associations between dementia and clinical, social, and nutritional factors. Dementia was diagnosed using a two-step procedure by physicians according to DSM-IV criteria. The prevalence of questionable dementia was 11.5% and definite dementia was 3.6%. Dementia prevalence increased with increasing years of age (adjusted OR: 1.04; 95% CI = 1.002-1.1) and decreased with more years of education (adjusted OR: 0.8; 95% CI = 0.6-0.99). Being malnourished increased the odds of dementia almost six-fold (adjusted OR: 5.9; 95% CI = 1.3-26.3), while frequent participation in social activities was associated with a decreased odds (adjusted OR: 0.5; 95% CI = 0.2-0.9). The prevalence of dementia in rural Bangladesh is similar to other countries in the South Asia region, but lower than reports from other world regions. Malnutrition is strongly associated with dementia occurrence, and is a relevant area for future research within low-income countries.


Structure-based design of Taq DNA polymerases with improved properties of dideoxynucleotide incorporation.
The Taq DNA polymerase is the most commonly used enzyme in DNA sequencing. However, all versions of Taq polymerase are deficient in two respects: (i) these enzymes incorporate each of the four dideoxynucleoside 5' triphosphates (ddNTPs) at widely different rates during sequencing (ddGTP, for example, is incorporated 10 times faster than the other three ddNTPs), and (ii) these enzymes show uneven band-intensity or peak-height patterns in radio-labeled or dye-labeled DNA sequence profiles, respectively. We have determined the crystal structures of all four ddNTP-trapped closed ternary complexes of the large fragment of the Taq DNA polymerase (Klentaq1). The ddGTP-trapped complex structure differs from the other three ternary complex structures by a large shift in the position of the side chain of residue 660 in the O helix, resulting in additional hydrogen bonds being formed between the guanidinium group of this residue and the base of ddGTP. When Arg-660 is mutated to Asp, Ser, Phe, Tyr, or Leu, the enzyme has a marked and selective reduction in ddGTP incorporation rate. As a result, the G track generated during DNA sequencing by these Taq polymerase variants does not terminate prematurely, and higher molecular-mass G bands are detected. Another property of these Taq polymerase variants is that the sequencing patterns produced by these enzymes are remarkably even in band-intensity and peak-height distribution, thus resulting in a significant improvement in the accuracy of DNA sequencing.


Using the lives saved tool (LiST) to model mHealth impact on neonatal survival in resource-limited settings.
While the importance of mHealth scale-up has been broadly emphasized in the mHealth community, it is necessary to guide scale up efforts and investment in ways to help achieve the mortality reduction targets set by global calls to action such as the Millennium Development Goals, not merely to expand programs. We used the Lives Saved Tool (LiST)--an evidence-based modeling software--to identify priority areas for maternal and neonatal health services, by formulating six individual and combined interventions scenarios for two countries, Bangladesh and Uganda. Our findings show that skilled birth attendance and increased facility delivery as targets for mHealth strategies are likely to provide the biggest mortality impact relative to other intervention scenarios. Although further validation of this model is desirable, tools such as LiST can help us leverage the benefit of mHealth by articulating the most appropriate delivery points in the continuum of care to save lives.


Creating Anti-icing Surfaces via the Direct Immobilization of Antifreeze Proteins on Aluminum.
Cryoprotectants such as antifreeze proteins (AFPs) and sugar molecules may provide a solution for icing problems. These anti-icing substances protect cells and tissues from freezing by inhibiting ice formation. In this study, we developed a method for coating an industrial metal material (aluminum, Al) with AFP from the Antarctic marine diatom, Chaetoceros neogracile (Cn-AFP), to prevent or delay ice formation. To coat Al with Cn-AFP, we used an Al-binding peptide (ABP) as a conjugator and fused it with Cn-AFP. The ABP bound well to the Al and did not considerably change the functional properties of AFP. Cn-AFP-coated Al (Cn-AFP-Al) showed a sufficiently low supercooling point. Additional trehalose coating of Cn-AFP-Al considerably delayed AFP denaturation on the Al without affecting its antifreeze activity. This metal surface-coating method using trehalose-fortified AFP can be applied to other metals important in the aircraft and cold storage fields where anti-icing materials are critical.


An association between the acute phase response and patterns of antigen induced T cell proliferation in juvenile idiopathic arthritis.
The aim of this research was to determine whether all memory T cells have the same propensity to migrate to the joint in patients with juvenile idiopathic arthritis. Paired synovial fluid and peripheral blood mononuclear cell proliferative responses to a panel of antigens were measured and the results correlated with a detailed set of laboratory and clinical data from 39 patients with juvenile idiopathic arthritis. Two distinct patterns of proliferative response were found in the majority of patients: a diverse pattern, in which synovial fluid responses were greater than peripheral blood responses for all antigens tested; and a restricted pattern, in which peripheral blood responses to some antigens were more vigorous than those in the synovial fluid compartment. The diverse pattern was generally found in patients with a high acute phase response, whereas patients without elevated acute phase proteins were more likely to demonstrate a restricted pattern. We propose that an association between the synovial fluid T cell repertoire and the acute phase response suggests that proinflammatory cytokines may influence recruitment of memory T cells to an inflammatory site, independent of their antigen specificity. Additionally, increased responses to enteric bacteria and the presence of alphaEbeta7 T cells in synovial fluid may reflect accumulation of gut associated T cells in the synovial compartment, even in the absence of an elevated acute phase response. This is the first report of an association between the acute phase response and the T cell population recruited to an inflammatory site.


Algorithm for the evaluation of imperfections in auto bodywork using profiles from a retroreflective image.
Nowadays the automobile industry is becoming more and more demanding as far as quality is concerned. Within the wide variety of processes in which this quality must be ensured, those regarding the squeezing of the auto bodywork are especially important due to the fact that the quality of the resulting product is tested manually by experts, leading to inaccuracies of all types. In this paper, an algorithm is proposed for the automated evaluation of the imperfections in the sheets of the bodywork after the squeezing process. The algorithm processes the profile signals from a retroreflective image and characterizes an imperfection. It is based on a convergence criterion that follows the line of the maximum gradient of the imperfection and gives its geometrical characteristics as a result: maximum gradient, length, width, and area.


Pertussis toxin treatment blocks hyperpolarization by muscarinic agonists in chick atrium.
Atrial and ventricular adenylate cyclase activity and atrial membrane potentials were measured in hearts from hatched chicks at 2-3 days after intravenous administration of pertussis toxin (0.5-1.0 micrograms, total) or saline. Both in atrium and ventricle, treatment with pertussis toxin antagonized inhibition by carbachol of basal and isoproterenol-stimulated adenylate cyclase activity without changing either basal or isoproterenol-stimulated adenylate cyclase. In atria from pertussis toxin-treated animals (5.4 mM potassium), carbachol hyperpolarized the resting membrane by 0.3 +/- 0.3 mV (n = 9) and did not increase resting potassium conductance. In contrast, carbachol hyperpolarized the resting membrane by 4.5 +/- 0.8 mV (n = 11) and increased resting potassium conductance more than 4-fold in saline-treated animals. Carbachol did not significantly affect the atrial action potential peak or duration at 50% repolarization of pertussis toxin-treated animals. This muscarinic agonist reduced action potential peak by 7.8 +/- 1.2 mV and the duration at 50% repolarization by 22.1 +/- 3.0 msec in atria from saline-treated animals. Pertussis toxin treatment also prevented the negative inotropic effect and the inhibition of calcium-dependent action potentials caused by carbachol in atrial muscle. Neither the affinity nor the maximal specific binding of [3H]quinuclidinyl benzilate in ventricular homogenates was changed by pertussis toxin treatment. The apparent affinity of carbachol for muscarinic receptor was slightly (approximately 2-fold) diminished in pertussis toxin-treated animals. The inhibition of carbachol-induced hyperpolarization by pertussis toxin treatment implicates a guanosine 5'-triphosphate-dependent protein (Ni or a similar protein) as an essential link that permits muscarinic receptor to regulate atrial potassium channels.


Considerations and Implications in the Purification of Extracellular Vesicles - A Cautionary Tale.
Extracellular vesicles (EVs) are nano-sized particles constitutively released from cells into all biological fluids. Interestingly, these vesicles contain genetic cargoes including proteins, RNA and bioactive lipids that can be functionally delivered and affect recipient cells. As a result, there is growing interest in studying EVs in pathological conditions, including central nervous system (CNS)-related diseases, as EVs may be used for diagnostic purposes or as therapeutic agents. However, one major bottleneck is the need for better EV purification strategies when considering complex biological sources such as serum/protein-rich media or plasma. In this study, we have performed a systematic comparison study between the current gold-standard method: ultracentrifugation, to an alternative: size-exclusion chromatography (LC), using induced pluripotent stem cell (iPSC) derived complex media as a model system. We demonstrate that LC allows for derivation of purer EVs from iPSCs, which was previously impossible with the original UC method. Importantly, our study further highlights the various drawbacks when using the conventional UC approach that lead to misinterpretation of EV data. Lastly, we describe novel data on our iPSC-EVs; how they could relate to stem cell biology and discuss their potential use as EV therapeutics for CNS diseases.


Management of severe malaria.
The case fatality of WHO-defined 'severe falciparum malaria' remains unacceptably high, at 10-20%. However, a gradual decline in case fatality in adults and children treated in hospitals may reflect use of improved regimens of antimalarial chemotherapy and increased awareness of important complications of the disease. The development of severe, perhaps inevitably-fatal, malaria might be prevented by early appropriate chemotherapy of uncomplicated disease. At the most peripheral levels of the health service, suppository formulations of artemisinin derivatives can be administered even to patients who are vomiting or prostrated. At dispensaries, clinics or hospitals, where intramuscular or intravenous administration of antimalarial drugs is possible, quinine and artemisinin derivatives are the treatments of choice. There is growing evidence of the safety and efficacy of the quinine loading dose and of the use of artemether and artesunate, based on large, randomised, controlled clinical studies. No safe and effective form of prophylactic ancillary treatment has yet emerged. Results of studies of antipyretics, anticonvulsants (phenobarbitone), anticytokine/anti-inflammatory agents (anti-TNF antibodies, pentoxifylline, dexamethasone), iron chelators and hyperimmune sera have been disappointing. Only blood transfusion and treatment of respiratory, circulatory and renal failure are of obvious benefit. New ideas are needed, based on what is known of the pathophysiology of severe disease.


Aggregatibacter aphrophilus pyogenic liver abscess in an immunocompetent young woman.
Aggregatibacter aphrophilus (formerly Haemophilus aphrophilus/paraphrophilus) is a small Gram-negative coccobacillus with fastidious growth requirements. It is a normal commensal of the human oropharynx and upper respiratory tract, and it can infrequently cause invasive human diseases, including bone and joint infections and subacute infective endocarditis. Cases of liver abscess caused by Aggregatibacter aphrophilus have been sparsely recorded in the English-language literature, but have not yet been reported in Taiwan. Here we present a case of Aggregatibacter aphrophilus pyogenic liver abscess in an immunocompetent young woman. She recovered uneventfully after repeated percutaneous abscess aspiration and antibiotic treatment for 5 weeks.


A phylogeny of caenorhabditis reveals frequent loss of introns during nematode evolution.
Since introns were discovered 26 years ago, people have wondered how changes in intron/exon structure occur, and what role these changes play in evolution. To answer these questions, we have begun studying gene structure in nematodes related to Caenorhabditis elegans. As a first step, we cloned a set of five genes from six different Caenorhabditis species, and used their amino acid sequences to construct the first detailed phylogeny of this genus. Our data indicate that nematode introns are lost at a very high rate during evolution, almost 400-fold higher than in mammals. These losses do not occur randomly, but instead, favor some introns and do not affect others. In contrast, intron gains are far less common than losses in these genes. On the basis of the sequences at each intron site, we suggest that several distinct mechanisms can cause introns to be lost. The small size of C. elegans introns should increase the rate at which each of these types of loss can occur, and might account for the dramatic difference in loss rate between nematodes and mammals.


Integration of metabolic and gene regulatory networks modulates the C. elegans dietary response.
Expression profiles are tailored according to dietary input. However, the networks that control dietary responses remain largely uncharacterized. Here, we combine forward and reverse genetic screens to delineate a network of 184 genes that affect the C. elegans dietary response to Comamonas DA1877 bacteria. We find that perturbation of a mitochondrial network composed of enzymes involved in amino acid metabolism and the TCA cycle affects the dietary response. In humans, mutations in the corresponding genes cause inborn diseases of amino acid metabolism, most of which are treated by dietary intervention. We identify several transcription factors (TFs) that mediate the changes in gene expression upon metabolic network perturbations. Altogether, our findings unveil a transcriptional response system that is poised to sense dietary cues and metabolic imbalances, illustrating extensive communication between metabolic networks in the mitochondria and gene regulatory networks in the nucleus.


Impact of gender on patient preferences for technology-based behavioral interventions.
Technology-based interventions offer an opportunity to address high-risk behaviors in the emergency department (ED). Prior studies suggest behavioral health strategies are more effective when gender differences are considered. However, the role of gender in ED patient preferences for technology-based interventions has not been examined. The objective was to assess whether patient preferences for technology-based interventions varies by gender. This was a secondary analysis of data from a systematic survey of adult (≥18 years of age), English-speaking patients in a large urban academic ED. Subjects were randomly selected during a purposive sample of shifts. The iPad survey included questions on access to technology, preferences for receiving health information, and demographics. We defined "technology-based" as web, text message, e-mail, social networking, or DVD; "non-technology-based" was defined as in-person, written materials, or landline. We calculated descriptive statistics and used univariate tests to compare men and women. Gender-stratified multivariable logistic regression models were used to examine associations between other demographic factors (age, race, ethnicity, income) and technology-based preferences for information on specific risky behaviors. Of 417 participants, 45.1% were male. There were no significant demographic differences between men and women. Women were more likely to use computers (90.8% versus 81.9%; p=0.03), Internet (66.8% versus 59.0%; p=0.03), and social networks (53.3% versus 42.6%; p=0.01). 89% of men and 90% of women preferred technology-based formats for at least type of health information; interest in technology-based for individual health topics did not vary by gender. Concern about confidentiality was the most common barrier to technology-based use for both genders. Multivariate analysis showed that for smoking, depression, drug/alcohol use, and injury prevention, gender modified the relationship between other demographic factors and preference for technology-based health information; e.g., older age decreases interest in technology-based information for smoking cessation in women but not in men (aOR 0.96, 95% CI 0.93-0.99 versus aOR 1.00, 95% CI 0.97-1.03). Our findings suggest ED patients' gender may affect technology preferences. Receptivity to technology-based interventions may be a complex interaction between gender and other demographic factors. Considering gender may help target ED patient populations most likely to be receptive to technology-based interventions.


Evidence for a major gene for bone mineral density/content in human pedigrees identified via probands with extreme bone mineral density.
Bone mineral content (BMC) and/or bone mineral density (BMD, i.e. BMC scaled by bone size) are major determinants for osteoporosis, which is a serious health problem. The major determinant of variation in BMD/BMC is genetic. The few studies now available are inconsistent in the identification and/or even in the existence of major gene(s) for BMD/BMC. In 51 human pedigrees with 941 individuals (526 measured for phenotypes) identified via probands with extreme BMD values, we performed complex segregation analyses to test the existence of a genetic locus with a major effect on BMD/BMC variation. We analyzed BMD and BMC at the spine, hip and wrist jointly by employing, as the study phenotype, factor scores (FS) of the principle component that explains approximately 75% of the total BMD/BMC variation at the three sites. The results indicate that a major gene exists with a codominant effect that is responsible for approximately 16% of the FS variation when adjusted for significant effects of sex, body weight and age. A significant genotype-x-sex-x-age interaction was found, which may explain approximately 14% of the FS variation after adjusting for body weight. Testing of various models did not provide support for shared familial environmental effects but suggested the existence of residual polygenic effects, which may explain approximately 50% of the FS variation when adjusting for sex, body weight and age. This study indicates a promising aspect of studies to identify a major gene for BMD/BMC variation in our pedigrees identified via extreme probands.


Type C bovine botulism outbreak due to carcass contaminated non-acidified silage.
The first reported bovine botulism outbreak in Finland is described. Nine out of 90 cattle on a dairy farm died after being fed non-acidified silage contaminated by animal carcasses. Type C botulinum neurotoxin gene was detected in one heifer by polymerase chain reaction (PCR) and the neurotoxin was detected by the mouse bioassay. Clostridium botulinum type C was isolated from liver samples. The isolated strain was identified with amplified fragment length polymorphism (AFLP) analysis as group III C. botulinum. To our knowledge, this is the first time that a type C bovine botulism outbreak has been diagnosed by PCR and confirmed by subsequent isolation and AFLP identification of the disease strain. The importance of the acidification process in silage production to inhibit C. botulinum toxin production in silage and thus to prevent further botulism outbreaks is emphasized. Nevertheless, preformed toxin in the carcass is not destroyed by acid.


Cognitive enhancement, cheating, and accomplishment.
An ethics of enhancement should not rest on blanket judgments; it should ask us to distinguish between the kinds of activities we want to enhance. Both students and academics have turned to cognition-enhancing drugs in significant numbers--but is their enhancement a form of cheating? The answer should hinge on whether the activity subject to enhancement is zero-sum or non-zero-sum, and whether one is more concerned with excellence in process or excellence in outcome. Cognitive enhancement should be especially tolerated when the activities at stake are non-zero-sum and when the importance of process is outweighed by the importance of outcome. The use of cognition-enhancing drugs does not unnaturally cheapen accomplishments achieved under their influence; instead, cognitive enhancement is in line with well-established conceptions of collaborative authorship, which shift the locus of praise and blame from individual creators to the ultimate products of their efforts.


NiO and Co3O4 nanoparticles induce lung DTH-like responses and alveolar lipoproteinosis.
Lung exposure to metal oxide nanoparticles (NPs) comprising soluble metal haptens may produce T-helper cell type 1 (Th1)- and Th17-associated delayed-type hypersensitivity (DTH) responses and pulmonary alveolar proteinosis (PAP). In order to study this, haptenic metal oxide NPs (NiO, Co(3)O(4), Cr(2)O(3) and CuO) were instilled into the lungs of female Wistar rats, and the immunoinflammatory responses were assessed at 24 h and 4 weeks post-instillation. Primary culture of alveolar macrophages from Wistar rats was used to evaluate the effect of the NPs on the ability to clear surfactant. NiO NPs induced chronic interstitial inflammation and pro-inflammatory Th1 and Th17 immune responses characterised by increases in the cytokines monocyte chemotactic protein (MCP)-1/CCL2, interleukin (IL)-12 p40, interferon-γ and IL-17A, whilst similar pathological responses induced by Co(3)O(4) NPs were associated with increases in MCP-1/CCL2 and IL-12 p40. However, neither Cr(2)O(3) nor CuO NPs elicited immunoinflammatory reactions. PAP was induced by both NiO and Co(3)O(4) NPs during the chronic phase. PAP was associated with over-production of surfactant by proliferation of type II cells and impaired clearance of surfactant by macrophages. These findings have implications for the risk management of occupational NP exposure and provide evidence that haptenic metal oxide NPs can induce chronic progressive lung immune responses via a DTH-like mechanism.


Chronic tibiofibular syndesmosis injury of ankle: evaluation with contrast-enhanced fat-suppressed 3D fast spoiled gradient-recalled acquisition in the steady state MR imaging.
To retrospectively determine the accuracy of coronal contrast material-enhanced fat-suppressed three-dimensional (3D) fast spoiled gradient-recalled acquisition in the steady state (SPGR) magnetic resonance (MR) imaging, as compared with that of routine transverse MR imaging, in the assessment of distal tibiofibular syndesmosis injury, with arthroscopy as the reference standard. The review board of the College of Medicine in Yonsei University approved this study; informed consent was waived. The study group comprised 45 patients (26 men, 19 women; mean age, 32.1 years; range, 18-58 years) with a chronic ankle injury who had undergone MR imaging and arthroscopic surgery. Three independent readers retrospectively reviewed the two sets of MR images (one set of gadolinium-enhanced 3D fast SPGR images and one set of routine T1-, T2-, and intermediate-weighted images). Scores from 1 to 5 in increasing order of the probability of injury were assigned to both sets. Arthroscopy was the reference standard. Syndesmotic recess height was measured on contrast-enhanced images. The two sets of images were compared for diagnostic performance with receiver operating characteristic (ROC) analysis. Dissection and histologic examination of six cadaveric ankles was performed to assess the syndesmotic area and ascertain the enhancing structure at MR imaging. At arthroscopy, syndesmotic injury was found in 24 ankles but not in 21 ankles. Areas under the ROC curve were significantly higher for the contrast-enhanced images (P<.05). The contrast-enhanced set showed higher accuracy, sensitivity, and specificity compared with the routine set for the assessment of syndesmosis injury. Mean syndesmotic recess height was significantly greater (P<.05) in patients with syndesmotic injury. Dissection and histologic examination revealed a highly vascular synovial fold in the syndesmotic area that is expected to enhance at MR imaging. In the assessment of chronic syndesmosis injury, coronal gadolinium-enhanced fat-suppressed 3D fast SPGR MR images were more sensitive, specific, and accurate than routine MR images.


Impact on health and provision of healthcare services during the COVID-19 lockdown in India: a multicentre cross-sectional study.
The COVID-19 pandemic resulted in a national lockdown in India from midnight on 25 March 2020, with conditional relaxation by phases and zones from 20 April. We evaluated the impact of the lockdown in terms of healthcare provisions, physical health, mental health and social well-being within a multicentre cross-sectional study in India. The SMART India study is an ongoing house-to-house survey conducted across 20 regions including 11 states and 1 union territory in India to study diabetes and its complications in the community. During the lockdown, we developed an online questionnaire and delivered it in English and seven popular Indian languages (Hindi, Tamil, Marathi, Telegu, Kannada, Bengali, Malayalam) to random samples of SMART-India participants in two rounds from 5 May 2020 to 24 May 2020. We used multivariable logistic regression to evaluate the overall impact on health and healthcare provision in phases 3 and 4 of lockdown in red and non-red zones and their interactions. A total of 2003 participants completed this multicentre survey. The bivariate relationships between the outcomes and lockdown showed significant negative associations. In the multivariable analyses, the interactions between the red zones and lockdown showed that all five dimensions of healthcare provision were negatively affected (non-affordability: OR 1.917 (95% CI 1.126 to 3.264), non-accessibility: OR 2.458 (95% CI 1.549 to 3.902), inadequacy: OR 3.015 (95% CI 1.616 to 5.625), inappropriateness: OR 2.225 (95% CI 1.200 to 4.126) and discontinuity of care: OR 6.756 (95% CI 3.79 to 12.042)) and associated depression and social loneliness. The impact of COVID-19 pandemic and lockdown on health and healthcare was negative. The exaggeration of income inequality during lockdown can be expected to extend the negative impacts beyond the lockdown.


Dual photonic-electrochemical lab on a chip for online simultaneous absorbance and amperometric measurements.
A dual lab on a chip (DLOC) approach that enables simultaneous optical and electrochemical detection working in a continuous flow regime is presented. Both detection modes are integrated for the first time into a single detection volume and operate simultaneously with no evidence of cross-talk. The electrochemical cell was characterized amperometrically by measuring the current in ferrocyanide solutions at +0.4 V vs gold pseudoreference electrode, at a flow rate of 200 μL min(-1). The experimental results for ferrocyanide concentrations ranging from 0.005 to 2 mM were in good agreement with the values predicted by the Levich equation for a microelectrode inside a rectangular channel, with a sensitivity of 2.059 ± 0.004 μA mM(-1) and a limit of detection (LoD) of (2.303 ± 0.004) × 10(-3) mM. Besides, optical detection was evaluated by measuring the absorbance of ferricyanide solutions at 420 nm. The results obtained therein coincide with those predicted by the Beer-Lambert law for a range of ferricyanide concentrations from 0.005 to 0.3 mM and showed an estimated LoD of (0.553 ± 0.001) × 10(-3) mM. The DLOC was finally applied to the analysis of L-lactate via a bienzymatic reaction involving lactate oxidase (LOX) and horseradish peroxidase (HRP). Here, the consumption of the reagent of the reaction (ferrocyanide) was continuously monitored by amperometry whereas the product of the reaction (ferricyanide) was recorded by absorbance. The DLOC presented good performance in terms of sensitivity and limit of detection, comparable to other fluidic systems found in the literature. Additionally, the ability to simultaneously quantify enzymatic reagent consumption and product generation confers the DLOC a self-verifying capability which in turn enhances its robustness and reliability.


Differences in reported medical conditions between myogenous and arthrogenous TMD patients and its relevance to the general practitioner.
The aim of this study was to compare self-reported medical conditions between a group of myogenous and arthrogenous temporomandibular disorder (TMD) patients and highlight its relevance to the general practitioner. The patient population consisted of 274 consecutive patients (14.6% male, 85.4% female; mean age 39.6 ± 14.2 years) diagnosed with either myogenous or arthrogenous TMD according to the Research Diagnostic Criteria for Temporomandibular Disorders. Self-reported medical conditions were derived from a standardized medical health questionnaire that patients completed during their initial examination. Data were compared between the two groups by means of chi-square tests, t tests and Mann-Whitney U tests. The level of significance was set at α = .05. Patients with myogenous TMD reported a greater number of medical conditions compared to arthrogenous TMD patients from the following broad categories: neurologic, gastrointestinal, musculoskeletal, psychologic, and "other". The following nine specific conditions were reported significantly more often in the myogenous group: severe headaches, fainting/dizzy spells, gastric acid reflux, fibromyalgia, anxiety, depression, psychiatric treatment, phobias, and frequent sore throats. The myogenous group reported pain to be significantly more severe than the arthrogenous group. Pain duration did not differ between the two groups. Patients with myogenous TMD self-reported significantly more comorbid disorders and more severe pain than patients with arthrogenous TMD. Understanding the differences between these two groups of patients will allow for more appropriate and targeted care for these populations. Future studies may focus on determining subgroups that are more likely to be indicative of a larger widespread pain syndrome to help guide individualized management strategies.


Some properties of the mammalian locomotory and respiratory systems in relation to body mass.
Video and ciné films of mammals running at the trot-gallop transition were analysed to measure breathing frequencies. Breathing frequency at the trot-gallop transition (fb, in Hz) was shown to decrease with increasing body mass (M, in kg) and was described by the equation fb = 5.08 M-0.14. The stiffness of the thorax and diaphragm of mice, rats, rabbits and wallabies was calculated and this, together with the mass of the viscera, was used to calculate the natural frequency of the system (nft, in Hz). The relationship between nft and body mass can be described by the equation nft = 5.02M-0.18. The significance of these results is discussed in relation to models of mechanical linkage between respiratory and locomotory movements.


New insights into repeat instability: role of RNA•DNA hybrids.
Expansion of tandem repeat sequences is responsible for more than 20 human diseases. Several cis elements and trans factors involved in repeat instability (expansion and contraction) have been identified. However no comprehensive model explaining large intergenerational or somatic changes of the length of the repeating sequences exists. Several lines of evidence, accumulated from different model studies, indicate that transcription through repeat sequences is an important factor promoting their instability. The persistent interaction between transcription template DNA and nascent RNA (RNA•DNA hybrids, R loops) was shown to stimulate genomic instability. Recently, we demonstrated that cotranscriptional RNA•DNA hybrids are preferentially formed at GC-rich trinucleotide and tetranucleotide repeat sequences in vitro as well as in human genomic DNA. Additionally, we showed that cotranscriptional formation of RNA•DNA hybrids at CTG•CAG and GAA•TTC repeats stimulate instability of these sequences in both E. coli and human cells. Our results suggest that persistent RNA•DNA hybrids may also be responsible for other downstream effects of expanded trinucleotide repeats, including gene silencing. Considering the extent of transcription through the human genome as well as the abundance of GC-rich and/or non-canonical DNA structure forming tandem repeats, RNA•DNA hybrids may represent a common mutagenic conformation. Hence, R loops are potentially attractive therapeutic target in diseases associated with genomic instability.


Detection of promoter methylation of tumor suppressor genes in serum DNA of breast cancer cases and benign breast disease controls.
Tumors are capable of shedding DNA into the blood stream. This shed DNA may be recovered from serum or plasma. The objective of this study was to evaluate whether pyrosequencing promoter DNA in a panel of 12 breast cancer-related genes (APC, BRCA1, CCND2, CDH1, ESR1, GSTP1, HIN1, P16, RARβ, RASSF1, SFRP1 and TWIST) to measure the degree of methylation would lead to a useful serum-based marker of breast cancer. Serum was obtained from women who were about to undergo a breast biopsy or mastectomy at three hospitals from 1977 to 1987 in Grand Rapids, MI USA. We compared the methylation status of 12 genes in serum DNA obtained from three groups of postmenopausal women (mean age at blood collection: 63.0 y; SD 9.9; range 35-91): breast cancer cases with lymph node-positive disease (n = 241); breast cancer cases with lymph node-negative disease (n = 63); and benign breast disease control subjects (n = 234). Overall, median levels of promoter methylation were low, typically below 5%, for all genes in all study groups. For all genes, median levels of methylation were higher (by 3.3 to 47.6%) in lymph node-positive breast cancer cases than in the controls. Comparing mean methylation level between lymph-node positive cases and controls, the most statistically significant findings, after adjustment of the false-positive rate (q-value), were for TWIST (p = 0.04), SFRP1 (p = 0.16), ESR1 (p = 0.17), P16 (p = 0.19) and APC (p = 0.19). For two of these four genes (TWIST, P16), the median methylation level was also highest in lymph-node positive cases, intermediate in lymph node-negative cases and lowest in the controls. The percent of study subjects with mean methylation scores ≥ 5% was higher among lymph node-positive cases than controls for ten genes, and significantly higher for HIN1 and TWIST (22.0 vs. 12.2%, p = 0.04 and 37.9 vs. 24.5%, p = 0.004, respectively). Despite relatively consistent variation in methylation patterns among groups, these modest differences did not provide sufficient ability to distinguish between cases and controls in a clinical setting.


Large-scale recovery of an endangered amphibian despite ongoing exposure to multiple stressors.
Amphibians are one of the most threatened animal groups, with 32% of species at risk for extinction. Given this imperiled status, is the disappearance of a large fraction of the Earth's amphibians inevitable, or are some declining species more resilient than is generally assumed? We address this question in a species that is emblematic of many declining amphibians, the endangered Sierra Nevada yellow-legged frog (Rana sierrae). Based on >7,000 frog surveys conducted across Yosemite National Park over a 20-y period, we show that, after decades of decline and despite ongoing exposure to multiple stressors, including introduced fish, the recently emerged disease chytridiomycosis, and pesticides, R. sierrae abundance increased sevenfold during the study and at a rate of 11% per year. These increases occurred in hundreds of populations throughout Yosemite, providing a rare example of amphibian recovery at an ecologically relevant spatial scale. Results from a laboratory experiment indicate that these increases may be in part because of reduced frog susceptibility to chytridiomycosis. The disappearance of nonnative fish from numerous water bodies after cessation of stocking also contributed to the recovery. The large-scale increases in R. sierrae abundance that we document suggest that, when habitats are relatively intact and stressors are reduced in their importance by active management or species' adaptive responses, declines of some amphibians may be partially reversible, at least at a regional scale. Other studies conducted over similarly large temporal and spatial scales are critically needed to provide insight and generality about the reversibility of amphibian declines at a global scale.


Effect of imidazole on renal function in unilateral ureteral-obstructed rat kidneys.
Imidazole has been proposed to reverse renal vasoconstriction following unilateral obstruction, presumably through blockade of thromboxane A2 (TXA2) synthesis. We examined this hypothesis in rats subjected to unilateral ureteral obstruction for 24 h by 1) performing renal function studies before and during imidazole infusion, and 2) measuring TXB2 and prostaglandin E2 (PGE2) in urine collected before and during imidazole infusion and the profile of products generated by metabolism of arachidonic acid with renal microsomes in vitro. Imidazole infusion was associated with only a bicarbonaturia in the postobstructed kidney; in contrast, clearance of PAH and inulin, fractional sodium excretion, and bicarbonate excretion were all increased in the contralateral kidney. In the postobstructed and contralateral kidneys, TXB2 excretion was diminished and PGE2 excretion was variable not only following imidazole infusion but after saline infusion as well. The profile of products generated by renal microsomal metabolism of arachidonic acid was similar among obstructed, contralateral, and normal kidneys. These results do not support the proposal that TXA2 is the mediator of renal vasoconstriction following unilateral ureteral obstruction.


A novel intraperitoneal metastatic xenograft mouse model for survival outcome assessment of esophageal adenocarcinoma.
Esophageal adenocarcinoma (EAC) has become the dominant type of esophageal cancer in United States. The 5-year survival rate of EAC is below 20% and most patients present with locally advanced or widespread metastatic disease, where current treatment is largely ineffective. Therefore, new therapeutic approaches are urgently needed. Improvement of EAC patient outcome requires well-characterized animal models in which to evaluate novel therapeutics. In this study we aimed to establish a peritoneal dissemination xenograft mouse model of EAC that would support survival outcome analyses. To find the best candidate cell line from 7 human EAC cell lines of different origin named ESO26, OE33, ESO51, SK-GT-2, OE19, OACM5.1C and Flo-1 were injected intraperitoneally/subcutaneously into SCID mice. The peritoneal/xenograft tumor formation and mouse survival were compared among different groups. All cell lines injected subcutaneously formed tumors within 3 months at variable rates. All cell lines except OACM5.1C formed intraperitoneal tumors within 3 months at variable rates. Median animal survival with peritoneal dissemination was 108 days for ESO26 cells (5X106), 65 days for OE33 cells (5X106), 88 days for ESO51 cells (5X106), 76 days for SK-GT-2 cells (5X106), 55 days for OE19 cells (5X106), 45 days for OE19 cells (10X106) and 82 days for Flo-1 cells (5X106). Interestingly, only in the OE19 model all mice (7/7 for 5X106 and 5/5 for10X106) developed bloody ascites with liver metastasis after intraperitoneal injection. The median survival time of these animals was the shortest (45 days for 10X106 cells). In addition, median survival was significantly increased after paclitaxel treatment compared with the control group (57 days versus 45 days, p = 0.0034) along with a significant decrease of the relative subcutaneous tumor volume (p = 0.00011). Thus peritoneal dissemination mouse xenograft model for survival outcome assessment after intraperitoneal injection of OE19 cells will be very useful for the evaluation of cancer therapeutics.


Energy Harvesting Using Thermocouple and Compressed Air.
In this paper, we describe the possibility of using the energy of a compressed air flow, where cryogenic temperatures are achieved within the flow behind the nozzle, when reaching a critical flow in order to maximize the energy gained. Compared to the energy of compressed air, the energy obtained thermoelectrically is negligible, but not zero. We are therefore primarily aiming to maximize the use of available energy sources. Behind the aperture separating regions with a pressure difference of several atmospheres, a supersonic flow with a large temperature drop develops. Based on the Seebeck effect, a thermocouple is placed in these low temperatures to create a thermoelectric voltage. This paper contains a mathematical-physical analysis for proper nozzle design, controlled gas expansion and ideal placement of a thermocouple within the flow for best utilization of the low temperature before a shockwave formation. If the gas flow passes through a perpendicular shockwave, the velocity drops sharply and the gas pressure rises, thereby increasing the temperature. In contrast, with a conical shockwave, such dramatic changes do not occur and the cooling effect is not impaired. This article also contains analyses for proper forming of the head shape of the thermocouple to avoid the formation of a detached shockwave, which causes temperature stagnation resulting in lower thermocouple cooling efficiency.


Hemocompatibility-Related Outcomes in the MOMENTUM 3 Trial at 6 Months: A Randomized Controlled Study of a Fully Magnetically Levitated Pump in Advanced Heart Failure.
The HeartMate 3 (HM3) Left Ventricular Assist System (LVAS) (Abbott) is a centrifugal, fully magnetically levitated, continuous-flow blood pump engineered to enhance hemocompatibility and reduce shear stress on blood components. The MOMENTUM 3 trial (Multicenter Study of MagLev Technology in Patients Undergoing Mechanical Circulatory Support Therapy With HeartMate 3) compares the HM3 LVAS with the HeartMate II (HMII) LVAS (Abbott) in advanced heart failure refractory to medical management, irrespective of therapeutic intention (bridge to transplant versus destination therapy). This investigation reported its primary outcome in the short-term cohort (n=294; 6-month follow-up), demonstrating superiority of the HM3 for the trial primary end point (survival free of a disabling stroke or reoperation to replace the pump for malfunction), driven by a reduced need for reoperations. The aim of this analysis was to evaluate the aggregate of hemocompatibility-related clinical adverse events (HRAEs) between the 2 LVAS. We conducted a secondary end point evaluation of HRAE (survival free of any nonsurgical bleeding, thromboembolic event, pump thrombosis, or neurological event) in the short-term cohort (as-treated cohort n=289) at 6 months. The net burden of HRAE was also assessed by using a previously described hemocompatibility score, which uses 4 escalating tiers of hierarchal severity to derive a total score for events encountered during the entire follow-up experience for each patient. In 289 patients in the as-treated group (151 the HM3 and 138 the HMII), survival free of any HRAE was achieved in 69% of the HM3 group and in 55% of the HMII group (hazard ratio, 0.62; confidence interval, 0.42-0.91; P=0.012). Using the hemocompatibility score, the HM3 group demonstrated less pump thrombosis requiring reoperation (0 versus 36 points, P<0.001) or medically managed pump thrombosis (0 versus 5 points, P=0.02), and fewer nondisabling strokes (6 versus 24 points, P=0.026) than the control HMII LVAS. The net hemocompatibility score in the HM3 in comparison with the HMII patients was 101 (0.67±1.50 points/patient) versus 137 (0.99±1.79 points/patient) (odds ratio, 0.64; confidence interval, 0.39-1.03; P=0.065). In this secondary analysis of the MOMENTUM 3 trial, the HM3 LVAS demonstrated greater freedom from HRAEs in comparison with the HMII LVAS at 6 months. URL: http://clinicaltrials.gov. Unique identifier: NCT02224755.


Non-invasive Markers of Liver Fibrosis: Adjuncts or Alternatives to Liver Biopsy?
Liver fibrosis reflects sustained liver injury often from multiple, simultaneous factors. Whilst the presence of mild fibrosis on biopsy can be a reassuring finding, the identification of advanced fibrosis is critical to the management of patients with chronic liver disease. This necessity has lead to a reliance on liver biopsy which itself is an imperfect test and poorly accepted by patients. The development of robust tools to non-invasively assess liver fibrosis has dramatically enhanced clinical decision making in patients with chronic liver disease, allowing a rapid and informed judgment of disease stage and prognosis. Should a liver biopsy be required, the appropriateness is clearer and the diagnostic yield is greater with the use of these adjuncts. While a number of non-invasive liver fibrosis markers are now used in routine practice, a steady stream of innovative approaches exists. With improvement in the reliability, reproducibility and feasibility of these markers, their potential role in disease management is increasing. Moreover, their adoption into clinical trials as outcome measures reflects their validity and dynamic nature. This review will summarize and appraise the current and novel non-invasive markers of liver fibrosis, both blood and imaging based, and look at their prospective application in everyday clinical care.


A prospective population-based cohort study of the prevalence, incidence and impact of obsessive-compulsive symptomatology.
Prospective longitudinal studies of obsessive-compulsive disorder (OCD) and sub-diagnostic obsessive-compulsive symptomatology in the non-clinical population, using age-defined cohorts, are rare. This study aimed to investigate the effect of OC symptoms on distress and psychosocial function and the effect of early-onset OC symptoms. 591 subjects drawn from the general population of Zurich, Switzerland were interviewed seven times between 1979 (at age 20/21) and 2008 (age 49/50). Data for socio-demographic variables and psychosocial impairment was also collected and compared with a control-group without OC symptoms drawn from the same population. The unweighted cumulative one-year rate of OCD in this sample was 5.1% and 21.7% reported some degree of clinically-relevant OC symptomatology (OCD or OCS). OCD appeared more prevalent in females whereas OCS and OC symptoms were more prevalent in males. The weighted cumulative prevalence rates, representative of the general population, for OCD, OCS and OC symptoms were 3.5%, 9.7%, and 11.2%, respectively. We could not identify OCD occurring before the age of 10 years, though sub-threshold cases were reported as early as age 2 years, whereas by the age of 22 years, around two thirds of OCD cases had emerged and no new cases developed OCD after around 37 years. Males were statistically significantly younger than females at onset of any OC symptomatology. Clinically-relevant OC syndromes start early and are associated with substantial distress, treatment-seeking activity and in the case of OCD, functional disability.


Active invasion of Porphyromonas gingivalis and infection-induced complement activation in ApoE-/- mice brains.
Periodontal disease is a polymicrobial inflammatory disease that leads to chronic systemic inflammation and direct infiltration of bacteria/bacterial components, which may contribute to the development of Alzheimer's disease. ApoE-/- mice were orally infected (n = 12) with Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Fusobacterium nucleatum as mono- and polymicrobial infections. ApoE-/- mice were sacrificed following 12 and 24 weeks of chronic infection. Bacterial genomic DNA was isolated from all brain tissues except for the F. nucleatum mono-infected group. Polymerase chain reaction was performed using universal 16 s rDNA primers and species-specific primer sets for each organism to determine whether the infecting pathogens accessed the brain. Sequencing amplification products confirmed the invasion of bacteria into the brain during infection. The innate immune responses were detected using antibodies against complement activation products of C3 convertase stage and the membrane attack complex. Molecular methods demonstrated that 6 out of 12 ApoE-/- mice brains contained P. gingivalis genomic DNA at 12 weeks (p = 0.006), and 9 out of 12 at 24 weeks of infection (p = 0.0001). Microglia in both infected and control groups demonstrated strong intracellular labeling with C3 and C9, due to on-going biosynthesis. The pyramidal neurons of the hippocampus in 4 out of 12 infected mice brains demonstrated characteristic opsonization with C3 activation fragments (p = 0.032). These results show that the oral pathogen P. gingivalis was able to access the ApoE-/- mice brain and thereby contributed to complement activation with bystander neuronal injury.


Prevalence of musculoskeletal disorders among dental healthcare providers: A systematic review and meta-analysis.
Background: Work-related musculoskeletal disorders (MSD) are common in dentistry due to the prolonged static work involved during patient care, making dental health care personnel vulnerable to musculoskeletal complaints. We aimed to pool the prevalence estimates of MSD among various dental healthcare providers, including dentists, dental students, dental hygienists, and auxiliaries. Methods: A systematic search of five databases was performed (Scopus, Embase, CINAHL, Web of Science, Dentistry & Oral Sciences Source). The studies that reported the prevalence of MSD among dental healthcare workers and those written in English were selected. Screening and data extraction were performed by two review authors independently. Discrepencies were resolved by another review author. Risk of bias assessment was done using a nine-item questionnaire developed by Hoy et al. Pooled estimates were calculated using meta-analysis of proportions (random effects model). Results: Among the 3090 publications screened, 234 publications were included for full-text screening. Meta-analysis was performed for 89 estimates from 88 publications. Females showed significantly higher prevalence [OR = 1.42 (95% CI = 1.09-1.84); I 2 = 66.02; N = 32]. The analysis yielded a pooled estimate of 78.4% (95% CI = 74.8-82). The meta-regression showed similar prevalence over the years (Coefficient: 0.001; P-value: 0.762). Conclusions: A high prevalence of MSD was noted among dental healthcare providers, with about seven out of ten having experienced MSD in the past. This emphasizes the need for awareness and adoption of appropriate ergonomic postures by dental healthcare providers from early in their careers to minimize work-related MSD.


The early nodulin gene MtN6 is a novel marker for events preceding infection of Medicago truncatula roots by Sinorhizobium meliloti.
MtN6 belongs to a series of cDNA clones representing Medicago truncatula genes transcriptionally activated during nodulation by Sinorhizobium meliloti (P. Gamas, F. de Carvalho Niebel, N. Lescure, and J. V. Cullimore, Mol. Plant-Microbe Interact. 9:233-242, 1996). We show here by in situ hybridization that MtN6 transcripts specifically accumulate first at very localized regions in the outer root cell layers, corresponding to outer cortical cells containing preinfection threads. At later stages, MtN6 expression is observed ahead of growing infection threads, including in the infection zone of mature root nodules. Interestingly, regulation of MtN6 is clearly distinct from that of other early nodulins expressed in the same region of the nodule, in terms of response to bacterial symbiotic mutants and to purified Nod factors. We thus suggest that MtN6 represents the first specific marker of a pathway involved in preparation to infection, which is at least partly controlled by Nod factors. Finally, we discuss the intriguing sequence homology shown by MtN6 to a protein from Emericella (Aspergillus) nidulans, FluG, that plays a key role in controlling the organogenesis of conidiophores (B. N. Lee and T. H. Adams, Genes Dev. 8:641-651, 1994).


Effects of initial overlap on the propagation of optical solitons at different wavelengths.
Effects of initial overlap between two solitons at different carrier wavelengths are studied theoretically and numerically as a function of the degree of overlap and the frequency separation of the two carriers. When the two solitons are fully overlapped, the carrier frequencies of the emerging two solitons are shifted toward each other by an amount proportional to the initial power and inversely proportional to the initial frequency separation with little effect on their amplitudes provided that the frequency separation is sufficiently larger than the spectral width of the solitons.


Trends in Irritable Bowel Syndrome Incidence among Taiwanese Adults during 2003-2013: A Population-Based Study of Sex and Age Differences.
No population-based irritable bowel syndrome (IBS) incidence data among Taiwanese adults are available. Whether IBS is associated with risk of organic colonic diseases remains unanswered. We investigated 1) the sex- and age-stratified trends in the annual incidence of IBS, and 2) the risk of selected organic diseases in patients with IBS compared with those without IBS among Taiwanese adults during 2003-2013. Medical claims data for 1 million randomly selected beneficiaries were obtained and analyzed. Patients with IBS were considered eligible for enrollment if they aged between 20 and 100 and had at least two medical encounters with IBS codes within 1 year. To test whether there was a linear secular trend in IBS incidence over time, multivariate Poisson regression with generalized estimating equation model was conducted. The risk of selected organic diseases associated with IBS was examined using multivariate Cox proportional hazard regression. From 2003 to 2013, the incidence of IBS significantly decreased over time [adjusted incidence rate ratio (IRR) = 0.97, p< 0.001]; the incidence of IBS significantly increased with age (adjusted IRR = 1.03, p < 0.001) and was significantly higher in women than in men (adjusted IRR = 1.14, p< 0.001). IBS significantly associated with increased risk of microscopic colitis, inflammatory bowel disease, and colorectal cancer during a 10-year follow-up period. The incidence of IBS increased with age and was slightly higher in women than in men among Taiwanese adults. During 2003-2013, IBS incidence gradually decreased over time. IBS may increase risk of several colonic organic diseases.


Rare cell isolation and analysis in microfluidics.
Rare cells are low-abundance cells in a much larger population of background cells. Conventional benchtop techniques have limited capabilities to isolate and analyze rare cells because of their generally low selectivity and significant sample loss. Recent rapid advances in microfluidics have been providing robust solutions to the challenges in the isolation and analysis of rare cells. In addition to the apparent performance enhancements resulting in higher efficiencies and sensitivity levels, microfluidics provides other advanced features such as simpler handling of small sample volumes and multiplexing capabilities for high-throughput processing. All of these advantages make microfluidics an excellent platform to deal with the transport, isolation, and analysis of rare cells. Various cellular biomarkers, including physical properties, dielectric properties, as well as immunoaffinities, have been explored for isolating rare cells. In this Focus article, we discuss the design considerations of representative microfluidic devices for rare cell isolation and analysis. Examples from recently published works are discussed to highlight the advantages and limitations of the different techniques. Various applications of these techniques are then introduced. Finally, a perspective on the development trends and promising research directions in this field are proposed.


Self-reconstructing all-optical poling in polymer fibers.
Self-sustained all-optical poling second-harmonic generation (SHG) experiments are conducted in single-core and multicore dye-doped poly(methyl methacrylate) optical fibers. By tuning the polarization of the fundamental beam, the SHG signal is degraded and is reconstructed spontaneously up to its initial level. We found a new situation in which the photo-induced self-organization of azo polymers creates a well-ordered periodic structure.


Ku is important for telomere maintenance, but not for differential expression of telomeric VSG genes, in African trypanosomes.
Trypanosome antigenic variation, involving differential expression of variant surface glycoprotein (VSG) genes, has a strong association with telomeres and with DNA recombination. All expressed VSGs are telomeric, and differential activation involves recombination into the telomeric environment or silencing/activation of subtelomeric promoters. A number of pathogen contingency gene systems associated with immune evasion involve telomeric loci, which has prompted speculation that chromosome ends provide conditions conducive for the operation of rapid gene switching mechanisms. Ku is a protein associated with eukaryotic telomeres that is directly involved in DNA recombination and in gene silencing. We have tested the hypothesis that Ku in trypanosomes is centrally involved in differential VSG expression. We show, via the generation of null mutants, that trypanosome Ku is closely involved in telomere length maintenance, more so for a transcriptionally active than an inactive telomere, but exhibits no detectable influence on DNA double strand break repair. The absence of Ku and the consequent great shortening of telomeres had no detectable influence either on the rate of VSG switching or on the silencing of the telomeric promoters of the VSG subset that is expressed in the tsetse fly.


Time-resolved renal contrast-enhanced MRA in normal dogs.
In humans, contrast-enhanced magnetic resonance angiography (CE-MRA) is a documented method to quantitatively and qualitatively evaluate renal vessels. It offers a safer alternative to computed tomography angiography. The aim of this study was to establish a renal MRA protocol in dogs using a 3D multiphase Fast Spoiled Gradient Recalled echo CE-MRA sequence (3D FSPGR CE-MRA). We used an elliptical centric ordering of k-space to acquire the contrast-sensitive data faster. Four to five consecutive 3D dorsal slabs, encompassing both kidneys, could be acquired in 40-65 s. The renal arterial and venous phases were obtained separately during phases 1 or 2, and 2 or 3, respectively without the need for any contrast medium bolus timing. The renal arteries and veins were clearly visualized. In conclusion, multiphase FSPGR 3D CE-MRA with an elliptical centric k-space ordering allows distinguishing renal arterial and venous phases in dogs, and can be used as a noninvasive diagnostic imaging alternative to map renal vessels in this species. Potential applications include the screening of renal donors in renal transplantation programs, and the pretreatment evaluation of animals with invasive renal neoplasia or renal vascular anomalies when surgery or embolization are contemplated.


[Effects of night shift on plasma concentrations of melatonin, LH, FSH and prolactin, and menstrual irregularity].
To examine the effect of night shift on the ovarian function, 122 teachers, 67 office workers, 377 nurses, 133 factory workers and 67 barmaids were surveyed. The incidence of irregular menstrual cycle was 13.1% in teachers, 14.9% in office workers, 24.9% in nurses, 36.8% in factory workers and 40.3% in barmaids. The incidence was significantly higher in women working at night than women working during the day. Plasma concentrations of melatonin, LH, FSH and prolactin were determined at 2200 h and 0200 h in 5 nurses working at night and in 6 nurses resting in their quarters. Plasma concentrations of melatonin and prolactin at 0200 h were significantly lower in nurses of the working group than others of the resting group, but plasma concentrations of LH and FSH did not differ between the two groups. These results indicate that night shift suppresses the ovarian function by affecting the circadian rhythm of melatonin and prolactin.


Advance care directive prevalence among older Australians and associations with person-level predictors and quality indicators.
Advance care planning (ACP) conversations may result in preferences for medical care being documented. To explore the uptake and quality of advance care directives (ACDs) among older Australians accessing health and aged care services, by overall ACP documentation prevalence, person-level predictors and ACD quality indicators. National multi-centre health record audit in general practices (GP), hospitals and residential aged care facilities (RACF). A total of 4187 people aged ≥65 years attending their GP (n = 676), admitted to hospital (n = 1122) or residing in a RACF (n = 2389). ACP documentation prevalence by setting and type including person-completed ACDs and non-ACD documents (completed by a health professional or someone else); person-level predictors and quality indicators of ACDs. Overall ACP documentation prevalence was 46.5% (29.2% weighted). ACD prevalence was 25.3% (14.2% weighted). Unweighted ACD prevalence was higher in RACFs (37.7%) than in hospitals (11.1%) and GPs (5.5%). 35.8% of ACP documentation was completed by a health professional (9.7% weighted), and 18.1% was completed by someone else (10.6% weighted). Having an ACD was positively associated with being female, older, having two or more medical conditions, receiving palliative care, being divorced/separated and being in a RACF. Only 73% of ACDs included full name, signature, document date and witnessing. Low ACP documentation prevalence and a lack of accessible, person-completed and quality ACDs represent an important ACP implementation issue. Low prevalence is complicated by poor document quality and a higher prevalence of documents being completed by someone other than the person.


Burnout Syndrome During COVID-19 Second Wave on ICU Caregivers.
The main objective of this article is to evaluate the prevalence of burnout syndrome (BOS) among the Intensive Care Unit (ICU) healthcare workers. The COVID-impact study is a study conducted in 6 French intensive care units. Five units admitting COVID patient and one that doesn't admit COVID patients. The survey was conducted between October 20th and November 20th, 2020, during the second wave in France. A total of 208 professionals responded (90% response rate). The Maslach Burnout Inventory scale, the Hospital Anxiety and Depression Scale and the Impact of Event Revisited Scale were used to study the psychological impact of the COVID-19 Every intensive care unit worker. The cohort includes 208 professionals, 52.4% are caregivers. Almost 20% of the respondents suffered from severe BOS. The professionals who are particularly affected by BOS are women, engaged people, nurses or reinforcement, not coming willingly to the intensive care unit and professionals with psychological disorders since COVID-19, those who are afraid of being infected, and people with anxiety, depression or post-traumatic stress disorder. Independent risk factors isolated were being engaged and being a reinforcement. Being a volunteer to come to work in ICU is protective. 19.7% of the team suffered from severe BOS during the COVID-19 pandemic in our ICU. The independent risk factors for BOS are: being engaged (OR = 3.61 (95% CI, 1.44; 9.09), don't working in ICU when it's not COVID-19 pandemic (reinforcement) (OR = 37.71 (95% CI, 3.13; 454.35), being a volunteer (OR = 0.10 (95% CI, 0.02; 0.46). Our study demonstrates the value of assessing burnout in health care teams. Prevention could be achieved by training personnel to form a health reserve in the event of a pandemic.


Effects of dilution on a six-element synthetic aperture.
An investigation was made of the effects of dilution on a six-element synthetic aperture. The characteristics of both the spread function and the transfer function were obtained for several degrees of dilution. It is shown that small degrees of dilution result in a significant gain over the performance of a single element, whereas more dilute apertures are inferior for direct imagery.


The evolution of autistic-like and schizotypal traits: a sexual selection hypothesis.
In this paper we present a new hypothesis on the evolution of autistic-like and schizotypal personality traits. We argue that autistic-like and schizotypal traits contribute in opposite ways to individual differences in reproductive and mating strategies, and have been maintained - at least in part - by sexual selection through mate choice. Whereas positive schizotypy can be seen as a psychological phenotype oriented to high-mating effort and good genes displays in both sexes, autistic-like traits in their non-pathological form contribute to a male-typical strategy geared toward high parental investment, low-mating effort, and long-term resource allocation. At the evolutionary-genetic level, this sexual selection hypothesis is consistent with Crespi and Badcock's "imprinted brain" theory of autism and psychosis; the effect of offspring mating behavior on resource flow within the family connects sexual selection with genomic imprinting in the context of human biparental care. We conclude by presenting the results of an empirical study testing one of the predictions derived from our hypothesis. In a sample of 199 college students, autistic-like traits predicted lower interest in short-term mating, higher partner-specific investment, and stronger commitment to long-term romantic relations, whereas positive schizotypy showed the opposite pattern of effects.


Diversity in antioxidant response enzymes in progressive stages of human nonalcoholic fatty liver disease.
Nonalcoholic fatty liver disease (NAFLD), which occurs in approximately 17 to 40% of Americans, encompasses progressive stages of liver damage ranging from steatosis to nonalcoholic steatohepatitis (NASH). Inflammation and oxidative stress are known characteristics of NAFLD; however, the precise mechanisms occurring during disease progression remain unclear. The purpose of the current study was to determine whether the expression or function of enzymes involved in the antioxidant response, NAD(P)H:quinone oxidoreductase 1 (NQO1), glutathione transferase (GST), and glutamate cysteine ligase, are altered in the progression of human NAFLD. Human livers staged as normal, steatotic, NASH (fatty), and NASH (not fatty) were obtained from the Liver Tissue Cell Distribution System. NQO1 mRNA, protein, and activity tended to increase with disease progression. mRNA levels of the GST isoforms A1, A2, A4, M3, and P1 increased with NAFLD progression. Likewise, GST A and P protein increased with progression; however, GST M protein levels tended to decrease. Of interest, total GST activity toward the substrate 1-chloro-2,4-dinitrobenzene decreased with NAFLD progression. GSH synthesis does not seem to be significantly dysregulated in NAFLD progression; however, the GSH/oxidized glutathione redox ratio seemed to be reduced with disease severity, indicating the presence of oxidative stress and depletion of GSH throughout progression of NAFLD. Malondialdehyde concentrations were significantly increased with disease progression, further indicating the presence of oxidative stress. Nuclear immunohistochemical staining of nuclear factor E2-related factor 2 (Nrf2), an indicator of activation of the transcription factor, was evident in all stages of NAFLD. The current data suggest that Nrf2 activation occurs in response to disease progression followed by induction of specific Nrf2 targets, whereas functionality of specific antioxidant defense enzymes seems to be impaired as NAFLD progresses.


Plasma renin activity in diabetes mellitus.
1. The plasma renin activity (PRA) was measured in 76 diabetic patients who were attending an outpatients clinic. Of these patients 16 had untreated hypertension and 28 had diabetic complications, which ranged from microaneurysms to renal failure and blindness. 2. Compared with age- and sex-matched normotensive control subjects, both normotensive and hypertensive diabetic patients had significantly higher PRA (P less than 0.001). 3. Hypertensive diabetic patients also showed a higher PRA than matched hypertensive control subjects (P less than 0.005). There were no significant differences between diabetic patients with hypertension or complications compared with those without these features. 4. Although this elevation of PRA could be due to a change in another component of the renin-angiotensin system, hypersecretion of renin is the most likely explanation.


Diagonal Acceleration for Covariance Matrix Adaptation Evolution Strategies.
We introduce an acceleration for covariance matrix adaptation evolution strategies (CMA-ES) by means of adaptive diagonal decoding (dd-CMA). This diagonal acceleration endows the default CMA-ES with the advantages of separable CMA-ES without inheriting its drawbacks. Technically, we introduce a diagonal matrix D that expresses coordinate-wise variances of the sampling distribution in DCD form. The diagonal matrix can learn a rescaling of the problem in the coordinates within a linear number of function evaluations. Diagonal decoding can also exploit separability of the problem, but, crucially, does not compromise the performance on nonseparable problems. The latter is accomplished by modulating the learning rate for the diagonal matrix based on the condition number of the underlying correlation matrix. dd-CMA-ES not only combines the advantages of default and separable CMA-ES, but may achieve overadditive speedup: it improves the performance, and even the scaling, of the better of default and separable CMA-ES on classes of nonseparable test functions that reflect, arguably, a landscape feature commonly observed in practice. The article makes two further secondary contributions: we introduce two different approaches to guarantee positive definiteness of the covariance matrix with active CMA, which is valuable in particular with large population size; we revise the default parameter setting in CMA-ES, proposing accelerated settings in particular for large dimension. All our contributions can be viewed as independent improvements of CMA-ES, yet they are also complementary and can be seamlessly combined. In numerical experiments with dd-CMA-ES up to dimension 5120, we observe remarkable improvements over the original covariance matrix adaptation on functions with coordinate-wise ill-conditioning. The improvement is observed also for large population sizes up to about dimension squared.


Identification of compound heterozygous KCNJ1 mutations (encoding ROMK) in a kindred with Bartter's syndrome and a functional analysis of their pathogenicity.
A multiplex family was identified with biochemical and clinical features suggestive of Bartter's syndrome (BS). The eldest sibling presented with developmental delay and rickets at 4 years of age with evidence of hypercalciuria and hypokalemia. The second sibling presented at 1 year of age with urinary tract infections, polyuria, and polydipsia. The third child was born after a premature delivery with a history of polyhydramnios and neonatal hypocalcemia. Following corrective treatment she also developed hypercalciuria and a hypokalemic metabolic alkalosis. There was evidence of secondary hyperreninemia and hyperaldosteronism in all three siblings consistent with BS. Known BS genes were screened and functional assays of ROMK (alias KCNJ1, Kir1.1) were carried out in Xenopus oocytes. We detected compound heterozygous missense changes in KCNJ1, encoding the potassium channel ROMK. The S219R/L220F mutation was segregated from father and mother, respectively. In silico modeling of the missense mutations suggested deleterious changes. Studies in Xenopus oocytes revealed that both S219R and L220F had a deleterious effect on ROMK-mediated potassium currents. Coinjection to mimic the compound heterozygosity produced a synergistic decrease in channel function and revealed a loss of PKA-dependent stabilization of PIP2 binding. In conclusion, in a multiplex family with BS, we identified compound heterozygous mutations in KCNJ1. Functional studies of ROMK confirmed the pathogenicity of these mutations and defined the mechanism of channel dysfunction.


Linking antimicrobial prescribing to antimicrobial resistance in the ICU: before and after an antimicrobial stewardship program.
Antimicrobials are an effective treatment for many types of infections, but their overuse promotes the spread of resistant microorganisms that defy conventional treatments and complicate patient care. In 2009, an antimicrobial stewardship program was implemented at Mount Sinai Hospital (MSH, Toronto, Canada). Components of this program were to alter the fraction of patients prescribed antimicrobials, to shorten the average duration of treatment, and to alter the types of antimicrobials prescribed. These components were incorporated into a mathematical model that was compared to data reporting the number of patients colonized with Pseudomonas aeruginosa and the number of patients colonized with antimicrobial-resistant P. aeruginosa first isolates before and after the antimicrobial stewardship program. Our analysis shows that the reported decrease in the number of patients colonized was due to treating fewer patients, while the reported decrease in the number of patients colonized with resistant P. aeruginosa was due to the combined effect of treating fewer patients and altering the types of antimicrobials prescribed. We also find that shortening the average duration of treatment was unlikely to have produced any noticeable effects and that further reducing the fraction of patients prescribed antimicrobials would most substantially reduce P. aeruginosa antimicrobial resistance in the future. The analytical framework that we derive considers the effect of colonization pressure on infection spread and can be used to interpret clinical antimicrobial resistance data to assess different aspects of antimicrobial stewardship within the ecological context of the intensive care unit.


Occupational therapy for community dwelling elderly people: a systematic review.
Occupational therapy might play an important role in maintaining independent living for community dwelling elderly people. The aim of this systematic review is to determine whether occupational therapy improves outcome for people who are >/=60 years and are living independently. An extensive search in MEDLINE, CINAHL, EMBASE, AMED and SCISEARCH until July 2002 was performed. Studies with controlled and uncontrolled designs were included. Six intervention categories were distinguished and individually analysed using a best-evidence synthesis, based on the type of design, the methodological quality, type of outcome measures and statistical significance of findings. 17 studies were included, ten of which were randomised clinical trials. Six randomised clinical trials had a high methodological quality. Strong evidence is present for the efficacy of advising on assistive devices as part of a home hazards assessment on functional ability. There is some evidence for the efficacy of training of skills combined with a home hazard assessment in decreasing the incidence of falls in elderly people at high risk of falling. Some evidence is available for the efficacy of comprehensive occupational therapy on functional ability, social participation and quality of life. Insufficient evidence is present for the efficacy of counselling the primary caregiver of dementia patients about maintaining the patient's functional abilities. This review shows that occupational therapy interventions for community dwelling elderly people results in positive outcomes. Future research in the efficacy of occupational therapy in elderly patient groups such as people with dementia is recommended. Furthermore, research into tailoring interventions to the needs of elderly patients is recommended.


Effect of Posterior Dynamic Instrumentation on High-Intensity Zone in Lumbar Degenerative Disc Disease.
To investigate high-intensity zone (HIZ) changes after lumbar posterior dynamic instrumentation. Our study included 53 patients: 27 patients in group 1, 26 patients in group 2. All patients had one or two levels of degenerative disc disease with an HIZ confirmed by magnetic resonance imaging (MRI). Group 1 underwent one- or two-level dynamic lumbar posterior instrumentation. Group 2 was treated conservatively with an exercise program. Patients were evaluated using MRI, a numerical visual analog pain scale (VAS), and the Oswestry Disability Index (ODI) at baseline, at 1 year after surgery, and at final follow-up evaluation. The mean duration of follow-up was 49.3 months in group 1 and 47.19 months in group 2. The baseline VAS and ODI scores were similar for both groups. The mean VAS score of group 1 was significantly improved at 1 year after surgery and at final follow-up. The mean ODI value was lower in group 1 than in group 2 at 1 year and at final follow-up. Pfirrmann grades in group 1 significantly differed at 1 year and at final follow-up but did not change in group 2. The number of HIZs significantly decreased in from baseline to 1 year and from baseline to final follow-up in group 1 but did not differ in group 2. Dynamic lumbar stabilization systems are promising. Observations such as Pfirrmann grade improvements and disappearance of HIZs are concordant with improvements in VAS and ODI scores demonstrate that dynamic stabilization systems may provide an environment for regeneration.


Uranium exposure to the tropical duckweed Lemna aequinoctialis and pulmonate snail Amerianna cumingi: fate and toxicity.
The discharge of catchment-management water from the Ranger uranium (U) mine into Magela Creek upstream of the Ramsar-listed Magela Floodplain in Kakadu National Park is an important part of the mine's water-management system. Because U is one of the primary toxicants associated with this water, a receiving-water trigger value (TV), based on chronic toxicity data from five local native species, was derived for U. To strengthen the data set underpinning the derivation of the TV, the chronic toxicity of U to two additional tropical freshwater species, duckweed Lemna aequinoctialis (96-hour growth rate), and pulmonate gastropod, Amerianna cumingi (96-hour reproduction), was determined. The fate of U within the test systems was an important component of the study because analysis of U concentrations during the snail tests indicated that a substantial proportion of U (approximately 25%) was being lost from the test solutions when integrated during the entire test duration. Analysis of the snails and their food for U indicated that only a small proportion that was lost from solution was being taken up by the snails. Therefore, the majority of U that was lost was considered unavailable to the snails, and thus the exposure concentrations used to calculate the toxicity estimates were adjusted downward. Integrating the loss of U from the L. aequinoctialis test solutions over time showed that only a small proportion (6% to 13%) was lost during the test: Of that, almost half (2-5%) was taken up by the plants (constituting exposure). Uranium was only moderately toxic to L. aequinoctialis, with no observed-effect concentrations, lowest observed-effect concentrations, and inhibition concentrations causing 10% and 50% effects (IC10 and IC50) values of 226, 404, 207, and 1435 microg/l, respectively. A. cumingi was found to be more sensitive to U than L. aequinoctialis, with NOEC, LOEC, IC10, and IC50 values of 60, 61, 15, and 278 microg/l, respectively. The data for these two additional species will be used to revise the current TV for U in Magela Creek.


An antibody that locks HER3 in the inactive conformation inhibits tumor growth driven by HER2 or neuregulin.
HER2/HER3 dimerization resulting from overexpression of HER2 or neuregulin (NRG1) in cancer leads to HER3-mediated oncogenic activation of phosphoinositide 3-kinase (PI3K) signaling. Although ligand-blocking HER3 antibodies inhibit NRG1-driven tumor growth, they are ineffective against HER2-driven tumor growth because HER2 activates HER3 in a ligand-independent manner. In this study, we describe a novel HER3 monoclonal antibody (LJM716) that can neutralize multiple modes of HER3 activation, making it a superior candidate for clinical translation as a therapeutic candidate. LJM716 was a potent inhibitor of HER3/AKT phosphorylation and proliferation in HER2-amplified and NRG1-expressing cancer cells, and it displayed single-agent efficacy in tumor xenograft models. Combining LJM716 with agents that target HER2 or EGFR produced synergistic antitumor activity in vitro and in vivo. In particular, combining LJM716 with trastuzumab produced a more potent inhibition of signaling and cell proliferation than trastuzumab/pertuzumab combinations with similar activity in vivo. To elucidate its mechanism of action, we solved the structure of LJM716 bound to HER3, finding that LJM716 bound to an epitope, within domains 2 and 4, that traps HER3 in an inactive conformation. Taken together, our findings establish that LJM716 possesses a novel mechanism of action that, in combination with HER2- or EGFR-targeted agents, may leverage their clinical efficacy in ErbB-driven cancers.


Significant Differences Characterise the Correlation Coefficients between Biocide and Antibiotic Susceptibility Profiles in Staphylococcus aureus.
There is a growing concern by regulatory authorities for the selection of antibiotic resistance caused by the use of biocidal products. We aimed to complete the detailed information on large surveys by investigating the relationship between biocide and antibiotic susceptibility profiles of a large number of Staphylococcus aureus isolates using four biocides and antibiotics commonly used in clinical practice. The minimal inhibitory concentration (MIC) for most clinically-relevant antibiotics was determined according to the standardized methodology for over 1600 clinical S. aureus isolates and compared to susceptibility profiles of benzalkonium chloride, chlorhexidine, triclosan, and sodium hypochlorite. The relationship between antibiotic and biocide susceptibility profiles was evaluated using non-linear correlations. The main outcome evidenced was an absence of any strong or moderate statistically significant correlation when susceptibilities of either triclosan or sodium hypochlorite were compared for any of the tested antibiotics. On the other hand, correlation coefficients for MICs of benzalkonium chloride and chlorhexidine were calculated above 0.4 for susceptibility to quinolones, beta-lactams, and also macrolides. Our data do not support any selective pressure for association between biocides and antibiotics resistance and furthermore do not allow for a defined risk evaluation for some of the compounds. Importantly, our data clearly indicate that there does not involve any risk of selection for antibiotic resistance for the compounds triclosan and sodium hypochlorite. These data hence infer that biocide selection for antibiotic resistance has had so far a less significant impact than feared.


Factors Associated With Noncompletion During the Run-In Period Before Randomization and Influence on the Estimated Benefit of LCZ696 in the PARADIGM-HF Trial.
The 8442 patients randomized in the Prospective Comparison of Angiotensin Receptor-Neprilysin Inhibitor With an Angiotensin-Converting Enzyme Inhibitor to Determine Impact on Global Mortality and Morbidity in Heart Failure (PARADIGM-HF) trial, in which sacubitril/valsartan (LCZ696) reduced both death and HF hospitalization more than enalapril, were a subset of 10 521 patients entering sequential, single-blind run-in periods (enalapril 10 mg twice daily for 2 weeks followed by LCZ696 200 mg twice daily for 4 to 6 weeks) to ensure short-term tolerability of the 2 study medications. We identified the predictors of run-in noncompletion and estimated the implications of noncompletion for the overall study result. Patient factors associated with run-in noncompletion were defined in multivariable logistic regression models. The effectiveness of LCZ696 in a broader cohort approximating the full run-in population was estimated by weighting randomized patients according to the inverse probability of run-in completion; 2079 (19.8%) subjects discontinued the study during the run-in period, including 1102 (10.5%) during the enalapril phase and 977 (9.3%) during the LCZ696 phase. In multivariable models, lower systolic blood pressure, lower estimated glomerular filtration rate, higher N-terminal pro-B-type natriuretic peptide, and ischemic cause of heart failure were associated with higher risk for run-in noncompletion. Repeat analysis of the effect of randomized treatment giving greater weight to randomized patients resembling those who did not complete the run-in did not alter the hazard ratio favoring LCZ696 over enalapril for the primary end point of cardiovascular death or heart failure hospitalization, or the additional key end points of cardiovascular death and all-cause mortality. Patients with lower blood pressure, lower glomerular filtration rate, and more severe heart failure were at higher risk for noncompletion during the run-in period of PARADIGM-HF. Weighted analysis of key study outcomes accounting for the effect of run-in noncompletion did not alter the benefit of LCZ696 over enalapril. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01035255.


Turnover of beta 1- and beta 2-adrenergic receptors after down-regulation or irreversible blockade.
The turnover of beta 1- and beta 2-adrenergic receptors was measured after both isoproterenol-induced down-regulation and irreversible blockade of receptors. Changes in the density of receptors were quantified using the radioligands 125I-iodopindolol and 125I-iodocyanopindolol. Treatment of intact L6 myoblasts or C6 glioma cells with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) inactivated beta-adrenergic receptors on membranes prepared from these cells. At a concentration of 100 microM EEDQ, more than 90% of beta 1- and beta 2-adrenergic receptors were inactivated within 2 hr of treatment. Recovery of beta-adrenergic receptors on intact cells after inactivation by EEDQ required more than 24 hr and was prevented by cycloheximide, an inhibitor of protein synthesis. The kinetics of recovery of the density of receptors were analyzed in terms of a model that allows estimation of the rate constants for receptor appearance in and disappearance from the membrane, assuming that the rate of appearance of receptors is constant and the rate of disappearance of receptors is proportional to the number of receptors. Beta 2-Adrenergic receptors on L6 myoblasts were incorporated into the membrane at a rate of 28 fmol/mg of protein/hr and had a half-life of 12.6 hr. On C6 glioma cells, Beta 1- and beta 2-adrenergic receptors appeared at rates of 13.3 and 6.6 fmol/mg of protein/hr, respectively, with half-lives of 9.4 and 6.4 hr. Recovery of receptors on C6 cells after isoproterenol-induced down-regulation was inhibited by cycloheximide. The rate of recovery of beta 1- and beta 2-adrenergic receptors was reduced after treatment with isoproterenol for 8 hr when compared to recovery after treatment with EEDQ. The major effect of treatment with isoproterenol was a persistent decrease in the rate of appearance of beta 1- and beta 2-adrenergic receptors (rate of synthesis and insertion into the membrane after treatment with isoproterenol = 4.0 fmol/mg of protein/hr). Since treatment with isoproterenol did not alter the rate of cell division or total protein synthesis, the isoproterenol-induced alteration was probably a specific effect on the rate of synthesis of beta-adrenergic receptors.


Does a Water Protocol Improve the Hydration and Health Status of Individuals with Thin Liquid Aspiration Following Stroke? A Randomized Controlled Trial.
The benefit of water protocols for individuals with thin liquid aspiration remains controversial, with mixed findings from a small number of randomized controlled trials (RCTs). This study aimed to contribute to the evidence of the effectiveness of water protocols with a particular emphasis on health outcomes, especially hydration. An RCT was conducted with patients with known thin liquid aspiration post stroke randomized to receiving thickened liquids only or a water protocol. For the 14 participants in rehabilitation facilities whose data proceeded to analysis, there was no difference in the total amount of beverages consumed between the water protocol group (mean = 1103 ml per day, SD = 215 ml) and the thickened liquids only group (mean = 1103 ml, SD = 247 ml). Participants in the water protocol group drank on average 299 ml (SD 274) of water but offset this by drinking less of the thickened liquids. Their hydration improved over time compared with participants in the thickened liquids only group, but differences between groups were not significant. Twenty-one percent of the total sample was diagnosed with dehydration, and no participants in either group were diagnosed with pneumonia. There were significantly more diagnoses of urinary tract infection in the thickened liquids only group compared to the water protocol group (χ (2) = 5.091, p = 0.024), but no differences between groups with regard to diagnoses of dehydration (χ (2) = 0.884, p = 0.347) or constipation (χ (2) = 0.117, p = 0.733). The findings reinforce evidence about the relative safety of water protocols for patients in rehabilitation post stroke and provide impetus for future research into the potential benefits for hydration status and minimizing adverse health outcomes.