Momordica charantia and type 2 diabetes: from in vitro to human studies.
Type 2 diabetes is a growing health problem worldwide that is particularly severe in India and China. In these areas, bitter gourd (Momordica charantia) is a popular vegetable which is traditionally known to have health beneficial effects not only, but mainly, on diabetes. Bitter gourd could be a cheap possibility to help the poor in these and other countries to control their blood glucose levels. This review describes anti-diabetic effects of bitter gourd reported in the literature and discusses what still needs to be clarified for developing an evidence-based and safe use of the bitter gourd for diabetes. Analyses of bioactive compounds have shown that bitter gourd is rich in nutrients and phytochemicals of which some have anti-diabetic effects. Juices, powders, extracts, and isolated compounds have been tested in vitro and in vivo. Bitter gourd increases insulin secretion of the pancreas, decreases intestinal glucose uptake, and increases uptake and utilization of glucose in peripheral tissues. Although human studies with type 2 diabetics are weak in their design and/or results, some of the studies do indicate anti-diabetic effects in patients and safety for bitter gourd treatment in humans. In the future, well designed studies with rodents will help to understand what kind of bitter gourd variety, dosage, preparation, and duration of administration is optimal. Such results will help to design human studies which are necessary to prove the effectiveness of bitter gourd in patients.


Acute Undifferentiated Febrile Illness in Patients Presenting to a Tertiary Care Hospital in South India: Clinical Spectrum and Outcome.
Acute undifferentiated febrile illness (AUFI) may have similar clinical presentation, and the etiology is varied and region specific. This prospective observational study was conducted in a tertiary hospital in South India. All adult patients presenting with AUFI of 3-14 days duration were evaluated for etiology, and the differences in presentation and outcome were analyzed. The study cohort included 1258 patients. A microbiological cause was identified in 82.5% of our patients. Scrub typhus was the most common cause of AUFI (35.9%) followed by dengue (30.6%), malaria (10.4%), enteric fever (3.7%), and leptospirosis (0.6%). Both scrub typhus and dengue fever peaked during the monsoon season and the cooler months, whereas no seasonality was observed with enteric fever and malaria. The mean time to presentation was longer in enteric fever (9.9 [4.7] days) and scrub typhus (8.2 [3.2] days). Bleeding manifestations were seen in 7.7% of patients, mostly associated with dengue (14%), scrub typhus (4.2%), and malaria (4.6%). The requirement of supplemental oxygen, invasive ventilation, and inotropes was higher in scrub typhus, leptospirosis, and malaria. The overall mortality rate was 3.3% and was highest with scrub typhus (4.6%) followed by dengue fever (2.3%). Significant clinical predictors of scrub typhus were breathlessness (odds ratio [OR]: 4.96; 95% confidence interval [CI]: 3.38-7.3), total whole blood cell count >10,000 cells/mm3 (OR: 2.31; 95% CI: 1.64-3.24), serum albumin <3.5 g % (OR: 2.32; 95% CI: 1.68-3.2). Overt bleeding manifestations (OR: 2.98; 95% CI: 1.84-4.84), and a platelet count of <150,000 cells/mm3 (OR: 2.09; 95% CI: 1.47-2.98) were independent predictors of dengue fever. The similarity in clinical presentation and diversity of etiological agents demonstrates the complexity of diagnosis and treatment of AUFI in South India. The etiological profile will be of use in the development of rational guidelines for control and treatment of AUFI.


PineappleDB: an online pineapple bioinformatics resource.
A world first pineapple EST sequencing program has been undertaken to investigate genes expressed during non-climacteric fruit ripening and the nematode-plant interaction during root infection. Very little is known of how non-climacteric fruit ripening is controlled or of the molecular basis of the nematode-plant interaction. PineappleDB was developed to provide the research community with access to a curated bioinformatics resource housing the fruit, root and nematode infected gall expressed sequences. PineappleDB is an online, curated database providing integrated access to annotated expressed sequence tag (EST) data for cDNA clones isolated from pineapple fruit, root, and nematode infected root gall vascular cylinder tissues. The database currently houses over 5600 EST sequences, 3383 contig consensus sequences, and associated bioinformatic data including splice variants, Arabidopsis homologues, both MIPS based and Gene Ontology functional classifications, and clone distributions. The online resource can be searched by text or by BLAST sequence homology. The data outputs provide comprehensive sequence, bioinformatic and functional classification information. The online pineapple bioinformatic resource provides the research community with access to pineapple fruit and root/gall sequence and bioinformatic data in a user-friendly format. The search tools enable efficient data mining and present a wide spectrum of bioinformatic and functional classification information. PineappleDB will be of broad appeal to researchers investigating pineapple genetics, non-climacteric fruit ripening, root-knot nematode infection, crassulacean acid metabolism and alternative RNA splicing in plants.


Predictors of Recurrent Ischemic Stroke in Patients with Embolic Strokes of Undetermined Source and Effects of Rivaroxaban Versus Aspirin According to Risk Status: The NAVIGATE ESUS Trial.
Embolic stroke of undetermined source (ESUS) identifies patients with cryptogenic ischemic stroke presumed due to embolism from several unidentified sources. Among patients with recent ESUS, we sought to determine independent predictors of recurrent ischemic stroke during treatment with aspirin or rivaroxaban and to assess the relative effects of these treatments according to risk. Exploratory analyses of 7213 participants in the NAVIGATE ESUS international trial who were randomized to aspirin 100 mg/day or rivaroxaban 15 mg/day and followed for a median of 11 months, during which time there were 309 first recurrent ischemic strokes (4.6% per year). Baseline features were correlated with recurrent stroke by multivariate analysis. The 7 independent predictors of recurrent stroke were stroke or transient ischemic attack (TIA) prior to the qualifying stroke (hazard ratio [HR] 2.03 95% confidence internal [CI] 1.58-2.60), current tobacco user (HR 1.62, 95% CI 1.24-2.12), age (HR 1.02 per year increase, 95%CI 1.01-1.03), diabetes (HR 1.28, 95% CI 1.01-1.64), multiple acute infarcts on neuroimaging (HR 1.49, 95% CI 1.09-2.02), aspirin use prior to qualifying stroke (HR 1.34, 95% CI 1.02-1.70), and time from qualifying stroke to randomization (HR .98, 95% CI .97-.99). The rate of recurrent stroke rate was 2.6% per year for participants without any of these risk factors, and increased by an average of 45% for each independent predictor (P < .001). There were no significant interactions between treatment effects and independent stroke predictors or stroke risk status. In this large cohort of ESUS patients, several features including prior stroke or TIA, advanced age, current tobacco user, multiple acute infarcts on neuroimaging, and diabetes independently identified those with an increased risk of ischemic stroke recurrence. The relative effects of rivaroxaban and aspirin were similar across the spectrum of independent stroke predictors and recurrent stroke risk status.


Grandparent caregiving among rural African Americans in a community in the American South: challenges to health and wellbeing.
An increasing number of grandparents in rural USA are serving as primary caregivers for their grandchildren because of parental incarceration, addiction, joblessness, or illness. Low-income, African American women from the South are overrepresented in this growing population. There is a paucity of research exploring the challenges faced by rural grandparent caregivers, and past studies have not explicitly addressed the potential consequences of rural grandparent caregiving for health. The purpose of this qualitative study was to explore grandparent caregiving among rural, low-income, African American grandmothers in a community in the American South, and to identify challenges to health that arose in that context. McLeroy's social ecological model (SEM) was used to examine these challenges at multiple levels of influence. This qualitative interview-based study was conducted in a high-poverty community in rural Georgia. In-depth interviews were conducted with African American grandparent caregivers and key informants from local community-based organizations. A key informant assisted in identifying initial interview participants, and then snowball sampling was used to recruit additional participants. Interview questions were grouped under five domains (intrapersonal, interpersonal, community, organizational, and policy), according to the levels of the SEM. Iterative content analysis of interview transcripts was utilized. Transcripts were coded to identify text segments related to each domain of the SEM, which were grouped together for analysis by domain. Reflexive memo-writing aided in development of themes, and data quality was assessed using Lincoln and Guba's trustworthiness criteria. Rural African American grandparent caregivers faced a range of challenges to health. Direct physical challenges included chronic pain that interfered with sleep and daily functioning, mobility issues exacerbated by child care, and the pressure of managing their own medical conditions as well as their grandchildren's. Financial scarcity added to their vulnerability to poor health outcomes, especially when caregivers would forego purchase of medications or visits to the doctor because of expenses related to their grandchildren. In addition, lack of child care made health appointments and hospitalizations logistically difficult. Emotional strain was common as grandparent caregivers struggled to protect their grandchildren in communities where rates of drug use, HIV, and incarceration were high. Caregivers worried about their mortality and the related consequences for their grandchildren. Chronic stress, which is linked to a number of poor health outcomes, was self-reported by most rural grandparent caregivers. In this study, the challenges of rural grandparent caregiving among African American women posed multiple threats to health and wellbeing. Further research is needed, in different rural contexts and with different caregiver populations, to more thoroughly examine the health risks of grandparent caregiving. In addition, the development of multi-faceted interventions and programs will be critical to meeting the needs of rural grandparent caregivers. A few models for such programs exist, although resource shortfalls have often limited their impact.


Bioavailability of zinc from inorganic and organic sources for pigs fed corn-soybean meal diets.
Two experiments were conducted with pigs 1) to determine the effect of supplemental Zn on growth performance, bone Zn, and plasma Zn in pigs fed Zn-unsupplemented, corn-soybean meal diets and 2) to assess bioavailability of Zn from inorganic and organic Zn sources. In both experiments, weanling pigs were fed a diet with no supplemental Zn for 5 wk to deplete their Zn stores. In Exp. 1, 192 pigs were fed a corn-soybean meal diet (growing diet, 32 mg/kg of Zn; finishing diet, 27 mg/kg of Zn) supplemented with feed-grade ZnSO4.H2O to provide 0, 5, 10, 20, 40, and 80 mg/kg of supplemental Zn. Supplemental Zn did not affect weight gain, feed intake, or gain/feed during either the growing or the finishing period (P > .05). However, bone and plasma Zn concentrations increased linearly (P < .01) in response to supplemental Zn at dietary Zn levels between 27 mg/kg (basal) and 47 mg/kg (breakpoint). In Exp. 2, three levels of supplemental Zn from ZnSO4.H2O (0, 7.5, and 15 mg/kg of supplemental Zn) were used to construct a standard curve (metacarpal, coccygeal vertebrae, and plasma Zn concentrations regressed on supplemental Zn intake; R2 = .93, .89, and .82, respectively). From the standard curve, the bone and plasma Zn concentrations obtained from pigs fed 15 mg/kg of supplemental Zn from ZnO and 7.5 mg/kg of supplemental Zn from Zn-methionine (ZnMET) and Zn-lysine (ZnLYS) were used to calculate bioavailable Zn via multiple linear regression, slope-ratio analysis. The estimates of Zn bioavailability differed depending on which variable was used. Overall trends indicated the following rankings: ZnSO4.H2O > ZnMet > ZnO > ZnLys.


Ex vivo expansion and selection of human CD34+ peripheral blood progenitor cells after introduction of a mutated dihydrofolate reductase cDNA via retroviral gene transfer.
Retroviral gene transfer into human myeloid precursor cells allows introduction of marker genes as well as genes conferring resistance to chemotherapeutic drugs. We transduced a human mutant dihydrofolate reductase (DHFR) cDNA into CD34 antigen-positive peripheral blood cells from patients with breast or ovarian cancer obtained after treatment with chemotherapy and granulocyte colony-stimulating factor (G-CSF). This mutant DHFR has been shown to confer resistance to methotrexate (MTX) in murine bone marrow. We established a transduction protocol that permitted ex vivo expansion and selection of transduced early progenitor cells. The number of progenitor cells from transduced CD34-positive cells increased 50-fold after cytokine prestimulation with interleukin-1 (IL-1), c-kit ligand (KL; stem cell factor), and IL-3 and 2 weeks in liquid culture. Transduced colony-forming unit-granulocyte-macrophage (CFU-GM), assayed directly after the transduction procedure, were protected completely against 2 x 10(-8) mol/L MTX, a concentration that significantly reduced the CFU-GM detected in the control population. Gene transfer of the mutant DHFR led to a twofold selective advantage for a pre-CFU population after exposure to MTX in liquid culture (P < .001). Polybrene, in contrast with protamine, significantly inhibited the expansion of progenitors. The presence of proviral DNA was monitored by polymerase chain reaction (PCR) and was detected in greater than 80% of CFU-GM and ex vivo expanded pre-CFU. We have demonstrated that human hematopoietic precursor cells can be expanded extensively after retroviral gene transfer. The same population of early progenitors can be selected ex vivo with low-dose MTX. As long-term expression of transduced genes in human hematopoietic cells remains a problem in vivo, these results may have implications for future clinical trials, especially for the introduction of nonselectable genes.


High-resolution 3D volumetry versus conventional measuring techniques for the assessment of experimental lymphedema in the mouse hindlimb.
Secondary lymphedema is a common complication of cancer treatment characterized by chronic limb swelling with interstitial inflammation. The rodent hindlimb is a widely used model for the evaluation of novel lymphedema treatments. However, the assessment of limb volume in small animals is challenging. Recently, high-resolution three-dimensional (3D) imaging modalities have been introduced for rodent limb volumetry. In the present study we evaluated the validity of microcomputed tomography (μCT), magnetic resonance imaging (MRI) and ultrasound in comparison to conventional measuring techniques. For this purpose, acute lymphedema was induced in the mouse hindlimb by a modified popliteal lymphadenectomy. The 4-week course of this type of lymphedema was first assessed in 6 animals. In additional 12 animals, limb volumes were analyzed by μCT, 9.4 T MRI and 30 MHz ultrasound as well as by planimetry, circumferential length and paw thickness measurements. Interobserver correlation was high for all modalities, in particular for μCT analysis (r = 0.975, p < 0.001). Importantly, caliper-measured paw thickness correlated well with μCT (r = 0.861), MRI (r = 0.821) and ultrasound (r = 0.800). Because the assessment of paw thickness represents a time- and cost-effective approach, it may be ideally suited for the quantification of rodent hindlimb lymphedema.


Early detection of venous thromboembolism in patients with neuroepithelial tumor: efficacy of screening with serum D-dimer measurements and Doppler ultrasonography.
The efficacy of combined serum D-dimer level measurement and Doppler ultrasonography of the lower extremity was investigated for screening of venous thromboembolism (VTE) in patients with neuroepithelial tumor. Eighty-one patients with neuroepithelial tumor were prospectively studied. All patients underwent measurement of serum D-dimer levels and Doppler ultrasonography of the lower extremity. The serum D-dimer level was measured every week, and Doppler ultrasonography was performed two and two weeks after surgery, then every two weeks until discharge, or every two weeks for patients who did not undergo surgery. If the serum D-dimer level increased over 10.0 μg/ml, Doppler ultrasonography or computed tomography was performed to detect VTE. VTE occurred in 12 (14.8%) patients (seven males and five females; age 34-75, mean 59.0 years). Only one patient was symptomatic, whereas 11 patients identified by the screening were without symptoms. Five patients were treated with anticoagulant therapy, one with prophylactic inferior vena cava filter placement with anticoagulant therapy, and the other six were closely followed up without medication. No patient died of pulmonary embolism. Serial Doppler ultrasonography showed thrombus regression or organization and no thrombus extension. The maximum serum D-dimer value was significantly higher in patients with VTE than in those without VTE (mean 14.5 vs. 3.46 μg/ml, P < 0.001). The D-dimer cutoff value of 5.4 μg/ml could be used to identify VTE with 83% sensitivity and 84% specificity. The combination of sequential serum D-dimer measurement and Doppler ultrasonography of the lower extremity is an efficient and non-invasive procedure for identifying asymptomatic VTE in patients with neuroepithelial tumor.


Oxidation of a [Cu2S] complex by N2O and CO2: insights into a role of tetranuclearity in the CuZ site of nitrous oxide reductase.
Oxidation of a [Cu2(μ-S)] complex by N2O or CO2 generated a [Cu2(μ-SO4)] product. In the presence of a sulfur trap, a [Cu2(μ-O)] species also formed from N2O. A [Cu2(μ-CS3)] species derived from CS2 modeled initial reaction intermediates. These observations indicate that one role of tetranuclearity in the CuZ catalytic site of nitrous oxide reductase is to protect the crucial S2- ligand from oxidation.


Evaluation of a revised curriculum: a four-year qualitative study of student perceptions.
Following curricular revisions at the Virginia Commonwealth University School of Dentistry, this longitudinal study was designed to determine students' perceptions of their educational experience in the revised curriculum. A SWOT (Strengths, Weaknesses, Opportunities, and Threats) open-ended response questionnaire was administered to students in the class of 2011 (N=89) in January of each academic year, 2008 through 2011, followed by focus groups three months prior to graduation. The overall response rate for the questionnaire was 69 percent, and a total of fourteen students participated in four focus groups. Cumulatively, 1,382 responses (SWOT=984 and focus groups=398) were qualitatively analyzed, and five themes emerged: 1) early clinical experiences led to a perceived readiness for direct patient care; 2) the pace and organization of the revised condensed preclinical curriculum were perceived as hectic yet were appreciated as necessary preparation for patient care; 3) most faculty members were seen as committed to student learning, but a few were reported to have poor teaching skills and attitudes when interacting with students; 4) a perceived lack of patients led to fewer clinical experiences and a decrease in student confidence; and 5) some curricular content was seen to be redundant and irrelevant to future practice. The results indicate that the students were satisfied with aspects of their educational experience, suggesting the revised curriculum's preliminary success in meeting its goals of earlier patient care, a condensed preclinical curriculum, and a student-friendly environment. As the curriculum is adapted in response to student feedback, ongoing evaluation is necessary and should be complemented by other evaluation indicators such as faculty perceptions and student learning outcomes.


Children's use of local and global visual features for material perception.
Adults can rapidly recognize material properties in natural images, and children's performance in material categorization tasks suggests that this ability develops slowly during childhood. In the current study, we further examined the information children use to recognize materials during development by asking how the use of local versus global visual features for material perception changes in middle childhood. We recruited adults and 5- to 10-year-old children for three experiments that required participants to distinguish between shape-matched images of real and artificial food. Accurate performance in this task requires participants to distinguish between a wide range of material properties characteristic of each category, thus testing material perception abilities broadly. In two tasks, we applied distinct methods of image scrambling (block scrambling and diffeomorphic scrambling) to parametrically disrupt global appearance while preserving features in small spatial neighborhoods. In the third task, we used image blurring to parametrically disrupt local feature visibility. Our key question was whether or not participant age affected performance differently when local versus global appearance was disrupted. We found that although image blur led to disproportionately poorer performance in young children, this effect was reduced or absent when diffeomorphic scrambling was used. We interpret this outcome as evidence that the ability to recruit large-scale visual features for material perception may develop slowly during middle childhood.


Non-invasive detection of animal nerve impulses with an atomic magnetometer operating near quantum limited sensitivity.
Magnetic fields generated by human and animal organs, such as the heart, brain and nervous system carry information useful for biological and medical purposes. These magnetic fields are most commonly detected using cryogenically-cooled superconducting magnetometers. Here we present the first detection of action potentials from an animal nerve using an optical atomic magnetometer. Using an optimal design we are able to achieve the sensitivity dominated by the quantum shot noise of light and quantum projection noise of atomic spins. Such sensitivity allows us to measure the nerve impulse with a miniature room-temperature sensor which is a critical advantage for biomedical applications. Positioning the sensor at a distance of a few millimeters from the nerve, corresponding to the distance between the skin and nerves in biological studies, we detect the magnetic field generated by an action potential of a frog sciatic nerve. From the magnetic field measurements we determine the activity of the nerve and the temporal shape of the nerve impulse. This work opens new ways towards implementing optical magnetometers as practical devices for medical diagnostics.


Dynamic instability of microtubules requires dynamin 2 and is impaired in a Charcot-Marie-Tooth mutant.
Dynamin is a fission protein that participates in endocytic vesicle formation. Although dynamin was originally identified as a microtubule-binding protein, the physiological relevance of this function was unclear. Recently, mutations in the ubiquitously expressed dynamin 2 (dyn2) protein were found in patients with Charcot-Marie-Tooth (CMT) disease, which is an inherited peripheral neuropathy. In this study, we show that one of these mutations, 551Delta3, induces prominent decoration of microtubules with the mutant dyn2. Dyn2 was required for proper dynamic instability of microtubules, and this was impaired in cells expressing the 551Delta3 mutant, which showed a remarkable increase in microtubule acetylation, a marker of stable microtubules. Depletion of endogenous dyn2 with a small interfering RNA also resulted in the accumulation of stable microtubules. Furthermore, the formation of mature Golgi complexes, which depends on microtubule-dependent membrane transport, was impaired in both dyn2 knockdown cells and cells expressing the 551Delta3 mutant. Collectively, our results suggest that dyn2 regulates dynamic instability of microtubules, which is essential for organelle motility, and that this function may be impaired in CMT disease.


Estimating minutes of physical activity from the previous day physical activity recall: validation of a prediction equation.
This study was designed to develop a prediction algorithm that would allow the Previous Day Physical Activity Recall (PDPAR) to be equated with temporally matched data from an accelerometer. Participants (n = 121) from a large, school-based intervention wore a validated accelerometer and completed the PDPAR for 3 consecutive days. Physical activity estimates were obtained from PDPAR by totaling 30-minute bouts of activity coded as ≥4 METS. A regression equation was developed in a calibration sample (n = 91) to predict accelerometer minutes of moderate to vigorous physical activity (MVPA) from PDPAR bouts. The regression equation was then applied to a separate, holdout sample (n = 30) to evaluate the utility of the prediction algorithm. Gender and PDPAR bouts accounted for 36.6% of the variance in accelerometer MVPA. The regression model showed that on average boys obtain 9.0 min of MVPA for each reported PDPAR bout, while girls obtain 4.8 min of MVPA per bout. When applied to the holdout sample, predicted minutes of MVPA from the models showed good agreement with accelerometer minutes (r = .81). The prediction equation provides a valid and useful metric to aid in the interpretation of PDPAR results.


The use of an anaesthetic simulator to assess single-use laryngoscopy equipment.
To compare the view at simulated direct laryngoscopy obtained with a standard laryngoscope, with and without a disposable cover and a disposable laryngoscope blade. Randomized non-blinded comparison. The high-fidelity human patient simulator at the Scottish Clinical Simulation Centre. Thirty-two anaesthetists with between 11 months and 25 years of experience. A randomized comparison of ease of laryngoscopy with each laryngoscope option for simulated easy and difficult laryngoscopy. The best grade achievable at laryngoscopy (Cormack and Lehane grade) for each laryngoscope, for both easy and difficult laryngoscopy. For the easy setting, 34% (P = 0.001) of anaesthetists graded laryngoscopy more difficult with the covered laryngoscope, and 22% (P = 0.008) with the disposable laryngoscope considered laryngoscopy more difficult than with the standard laryngoscope. For the difficult simulator setting, 69% (P < 0.001) found laryngoscopy more difficult with the covered laryngoscope and 69% (P < 0.001) with the disposable laryngoscope, when compared with the standard laryngoscope. There was no difference between the laryngoscopy grades for the covered and disposable laryngoscope for either easy (P = 0.21) or difficult (P = 0.87) simulation. Single-use equipment, as presently recommended for tonsillectomy surgery by the UK Department of Health, makes laryngoscopy more difficult for anaesthetists.


Microwave-assisted self-doping of TiO2 photonic crystals for efficient photoelectrochemical water splitting.
In this article, we report that the combination of microwave heating and ethylene glycol, a mild reducing agent, can induce Ti(3+) self-doping in TiO2. A hierarchical TiO2 nanotube array with the top layer serving as TiO2 photonic crystals (TiO2 NTPCs) was selected as the base photoelectrode. The self-doped TiO2 NTPCs demonstrated a 10-fold increase in visible-light photocurrent density compared to the nondoped one, and the optimized saturation photocurrent density under simulated AM 1.5G illumination was identified to be 2.5 mA cm(-2) at 1.23 V versus reversible hydrogen electrode, which is comparable to the highest values ever reported for TiO2-based photoelectrodes. The significant enhancement of photoelectrochemical performance can be ascribed to the rational coupling of morphological and electronic features of the self-doped TiO2 NTPCs: (1) the periodically morphological structure of the photonic crystal layer traps broadband visible light, (2) the electronic interband state induced from self-doping of Ti(3+) can be excited in the visible-light region, and (3) the captured light by the photonic crystal layer is absorbed by the self-doped interbands.


The prognostic value of androgen receptor (AR) in HER2-enriched metastatic breast cancer.
The significance of androgen receptor (AR) in metastatic breast cancer (MBC) remains unclear, and it is still largely unknown how AR expression level influences HER2-positive tumors. This study aimed to investigate the prognostic and predictive value of AR in HER2-enriched MBC. Primary and/or paired metastatic tumors of 304 patients with pathologically confirmed HER2-enriched MBC were collected and immunohistochemically assessed for AR expression. The associations of AR and other clinicopathological characteristics were compared using the Chi-square test. Progression-free survival (PFS) and overall survival (OS) were calculated using the Kaplan-Meier method and log-rank test. Cox regression analysis was used to determine independent prognostic factors. AR-positivity with a cut-off value of 10% was observed in 237 (78.0%) cases and was associated with longer PFS, 13.2 months, as compared to that of 8.2 months (P = 0.004) in patients with AR-negativity. Moreover, a significant increase in the 5-year OS rate (65.3% vs 36.2%, P < 0.001) was also observed for patients with AR-positive tumors. Cox regression analysis identified AR-positivity as an independent prognostic factor of both PFS (hazard ratio = 0.71, P = 0.039) and OS (HR = 0.53, P = 0.013). Additionally, for those who received first-line Trastuzumab therapies, prolonged PFS (15.8 months vs 8.2 months, P = 0.005) and 5-year OS rate (66.2% vs 26.2%, P = 0.009) were observed in AR-positive tumors compared to AR-negative ones. In conclusion, AR was identified as an independent prognostic factor for favorable PFS and OS and could also predict the efficacy of first-line Trastuzumab treatment in patients with HER2-enriched MBC.


Primary care treatment of insomnia: study protocol for a pragmatic, multicentre, randomised controlled trial comparing nurse-delivered sleep restriction therapy to sleep hygiene (the HABIT trial).
Insomnia is a prevalent sleep disorder that negatively affects quality of life. Multicomponent cognitive-behavioural therapy (CBT) is the recommended treatment but access remains limited, particularly in primary care. Sleep restriction therapy (SRT) is one of the principal active components of CBT and could be delivered by generalist staff in primary care. The aim of this randomised controlled trial is to establish whether nurse-delivered SRT for insomnia disorder is clinically and cost-effective compared with sleep hygiene advice. In the HABIT (Health-professional Administered Brief Insomnia Therapy) trial, 588 participants meeting criteria for insomnia disorder will be recruited from primary care in England and randomised (1:1) to either nurse-delivered SRT (plus sleep hygiene booklet) or sleep hygiene booklet on its own. SRT will be delivered over 4 weekly sessions; total therapy time is approximately 1 hour. Outcomes will be collected at baseline, 3, 6 and 12 months post-randomisation. The primary outcome is self-reported insomnia severity using the Insomnia Severity Index at 6 months. Secondary outcomes include health-related and sleep-related quality of life, depressive symptoms, use of prescribed sleep medication, diary and actigraphy-recorded sleep parameters, and work productivity. Analyses will be intention-to-treat. Moderation and mediation analyses will be conducted and a cost-utility analysis and process evaluation will be performed. Ethical approval was granted by the Yorkshire and the Humber - Bradford Leeds Research Ethics Committee (reference: 18/YH/0153). We will publish our primary findings in high-impact, peer-reviewed journals. There will be further outputs in relation to process evaluation and secondary analyses focussed on moderation and mediation. Trial results could make the case for the introduction of nurse-delivered sleep therapy in primary care, increasing access to evidence-based treatment for people with insomnia disorder. ISRCTN42499563.


Light tunneling inhibition in array of couplers with longitudinal refractive index modulation.
We consider light tunneling inhibition in a periodic array of optical couplers due to the specially designed longitudinal and transverse modulation of the refractive index. We show that local out-of-phase longitudinal modulation of refractive index in channels of directional couplers in combination with the global refractive index modulation between adjacent couplers allow simultaneous suppression of both local and global energy tunneling inside each coupler and between adjacent couplers. This enables the localization of light in single waveguide despite the remarkable difference of corresponding local and global energy tunneling rates.


U.S. Adults' Participation in Specific Activities: Behavioral Risk Factor Surveillance System--2011.
There is little information on national estimates for participation in types of aerobic activities among U.S. adults. Current estimates are important to develop appropriate and effective interventions to promote physical activity and interpret bias for some activities measured with devices. The percentage of adults participating in specific aerobic activities was estimated overall and by demographic subgroups. The 2011 Behavioral Risk Factor Surveillance System respondents (N = 446,216) reported up to 2 aerobic activities they spent the most time doing during the past month. Overall, walking (47%) was the most common activity reported and was reported more by women (54%) than men (41%). Participation in most activities declined with increasing age (P < .006). There were a number of differences in participation between race/ethnic subgroups. Participation increased with more education (P for trend < 0.006) for all activities. Participation in most activities was different (P < .002) across BMI subgroups. Walking is the most common activity, overall and among most subgroups. Other activity profiles differ by demographic subgroup. Physical activity promotion strategies that focus on identifying and addressing personal and environmental barriers and understanding demographic subgroup differences could lead to more tailored interventions and public health programs.


Opposing roles of Dnmt1 in early- and late-stage murine prostate cancer.
Previous studies have shown that tumor progression in the transgenic adenocarcinoma of mouse prostate (TRAMP) model is characterized by global DNA hypomethylation initiated during early-stage disease and locus-specific DNA hypermethylation occurring predominantly in late-stage disease. Here, we utilized Dnmt1 hypomorphic alleles to examine the role of Dnmt1 in normal prostate development and in prostate cancer in TRAMP. Prostate tissue morphology and differentiation status was normal in Dnmt1 hypomorphic mice, despite global DNA hypomethylation. TRAMP; Dnmt1 hypomorphic mice also displayed global DNA hypomethylation, but were characterized by altered tumor phenotype. Specifically, TRAMP; Dnmt1 hypomorphic mice exhibited slightly increased tumor incidence and significantly increased pathological progression at early ages and, conversely, displayed slightly decreased tumor incidence and significantly decreased pathological progression at advanced ages. Remarkably, hypomorphic Dnmt1 expression abrogated local and distant site macrometastases. Thus, Dnmt1 has tumor suppressor activity in early-stage prostate cancer, and oncogenic activity in late stage prostate cancer and metastasis. Consistent with the biological phenotype, epigenomic studies revealed that TRAMP; Dnmt1 hypomorphic mice show dramatically reduced CpG island and promoter DNA hypermethylation in late-stage primary tumors compared to control mice. Taken together, the data reveal a crucial role for Dnmt1 in prostate cancer and suggest that Dnmt1-targeted interventions may have utility specifically for advanced and/or metastatic prostate cancer.


Effect of polyethylene glycol electrolyte balanced solution on patients with acute colonic pseudo obstruction after resolution of colonic dilation: a prospective, randomised, placebo controlled trial.
Conservative therapy for patients with acute colonic pseudo obstruction (Ogilvie's syndrome) may be successful initially but relapses are common. The aim of the present study was to evaluate the effect of polyethylene glycol (PEG) electrolyte balanced solution on the relapse rate of the syndrome after initial resolution with neostigmine or endoscopic decompression. The study was performed on 30 consecutive patients who presented with abdominal distension and radiographic evidence of colonic dilation, with a caecal diameter > or = 10 cm, that resolved conservatively. Patients then were randomised to receive daily 29.5 g of PEG (n = 15) or similar placebo (n = 15). Patients were monitored daily for a seven day period for stool and flatus evacuations, and colonic diameter on abdominal radiographs. Administration of the test solutions and assessment of patient symptoms and x rays were performed in a blinded fashion. A caecal diameter > or = 8 cm with a concomitant > or =10% increase after initial successful therapeutic intervention was considered as a relapse and these patients, after a second therapeutic intervention, were eligible to receive open label PEG. Twenty five patients received neostigmine as the initial therapeutic intervention which resulted in resolution of colonic dilation in 88% of cases. Eight patients had successful endoscopic decompression. Five (33.3%) patients in the placebo group had recurrent caecal dilation compared with none in the PEG group (p = 0.04). Therapy with PEG resulted in a significant increase in stool and flatus evacuations (p = 0.001 and 0.032, respectively) as well as in a significant decrease in the diameter of caecum, ascending and transverse colon, and abdominal circumference (p = 0.017, 0.018, 0.014, and 0.008, respectively). Administration of PEG in patients with Ogilvie's syndrome after initial resolution of colonic dilation may increase the sustained response rate after initial therapeutic intervention.


Familial breast cancer: characteristics and outcome of BRCA 1-2 positive and negative cases.
The clinical and pathological characteristics and the clinical course of patients with breast cancer and BRCA 1-2 mutation are poorly known. From 1997, patients with breast cancer and a family history of breast or ovarian cancer were offered BRCA testing. The clinical and pathological features of patients with known BRCA status were retrospectively assessed and comparisons were made between cancers arising in BRCA positive and BRCA wild type (WT) patients respectively. Type of treatment, pattern of relapse, event (local relapse, contralateral breast cancer, metastases) free and overall survival were also compared in the two groups. Out of the 210 patients tested, 125 had been treated and followed-up at our Institution and were evaluated in this study. BRCA positive patients tended to be more often premenopausal (79% vs 65%) and to have positive lymphnodes (63% vs 49%), poorly differentiated tumours (76% vs 40%--p = 0.002 at univariate analysis, not significant at multivariate analysis) and negative estrogen receptors (43% vs 29%). Treatment was not different in the two groups. In the 86 BRCA-WT patients, the first event was a local relapse in 3 (3%), metachronous contralateral breast cancer in 7 (8%) and distant metastases in 16 (19%). In the 39 BRCA positive patients, the corresponding figures were 3 (8%), 8 (21%) and 3 (8%). There was no difference in event free survival, with a median of 180 months in both groups of patients. At 20 years, projected survival was 85% for BRCA positive patients and 55% for BRCA-WT, but this difference was not statistically significant. Although BRCA positive patients have more frequently negative prognostic factors, their prognosis appears to be equal to or better than in patients with BRCA-WT.


Heavy drinking in early adulthood and outcomes at mid life.
Heavy drinking in early adulthood among Blacks, but not Whites, has been found to be associated with more deleterious health outcomes, lower labor market success and lower educational attainment at mid-life. This study analysed psychosocial pathways underlying racial differences in the impact of early heavy alcohol use on occupational and educational attainment at mid-life. Outcomes in labor market participation, occupational prestige and educational attainment were measured in early and mid-adulthood. A mixture model was used to identify psychosocial classes that explain how race-specific differences in the relationship between drinking in early adulthood and occupational outcomes in mid-life operate. Data came from Coronary Artery Risk Development in Young Adults, a longitudinal epidemiologic study. Especially for Blacks, heavy drinking in early adulthood was associated with a lower probability of being employed in mid-life. Among employed persons, there was a link between heavy drinking for both Whites and Blacks and decreased occupational attainment at mid-life. We grouped individuals into three distinct distress classes based on external stressors and indicators of internally generated stress. Blacks were more likely to belong to the higher distressed classes as were heavy drinkers in early adulthood. Stratifying the data by distress class, relationships between heavy drinking, race and heavy drinking-race interactions were overall weaker than in the pooled analysis. Disproportionate intensification of life stresses in Blacks renders them more vulnerable to long-term effects of heavy drinking.


Linearity of the Faraday-rotation-type ac magnetic-field sensor with a ferrimagnetic or ferromagnetic rotator film.
We analyze the linearity and modulation depth of ac magnetic-field sensors or current sensors, using a ferrimagnetic or ferromagnetic film as the Faraday rotator and employing the detection of only the zeroth-order optical diffraction component from the rotator. It is theoretically shown that for this class of sensor the condition of a constant modulation depth and that of a constant ratio error give an identical series of curves for the relationship between Faraday rotation angle Θ and polarizer/analyzer relative angle Φ. We give some numerical examples to demonstrate the usefulness of the result with reference to a rare-earth iron garnet film as the rotator.


A vasculature-associated niche for undifferentiated spermatogonia in the mouse testis.
Mammalian spermatogenesis produces numerous sperm for a long period based on a highly potent stem cell system, which relies on a special microenvironment, or niche, that has not yet been identified. In this study, using time-lapse imaging of green fluorescent protein-labeled undifferentiated spermatogonia (A(undiff)) and three-dimensional reconstitution, we revealed a biased localization of A(undiff) to the vascular network and accompanying Leydig and other interstitial cells, in intact testes. Differentiating spermatogonia left these niche regions and dispersed throughout the basal compartment of the seminiferous epithelium. Moreover, rearrangement of A(undiff) accompanied the vasculature alteration. We propose that the mammalian germline niche is established as a consequence of vasculature pattern formation. This is different from what is observed in Drosophila or Caenorhabditis elegans, which display developmentally specified niche structures within polarized gonads.


The Specificity of Rugby Union Training Sessions in Preparation for Match Demands.
Investigations into the specificity of rugby union training practices in preparation for competitive demands have predominantly focused on physical and physiological demands. The evaluation of the contextual variance in perceptual strain or skill requirements between training and matches in rugby union is unclear, yet holistic understanding may assist to optimize training design. This study evaluated the specificity of physical, physiological, perceptual, and skill demands of training sessions compared with competitive match play in preprofessional, elite club rugby union. Global positioning system devices, video capture, heart rate, and session ratings of perceived exertion were used to assess movement patterns, skill completions, physiologic, and perceptual responses, respectively. Data were collected across a season (training sessions n = 29; matches n = 14). Participants (n = 32) were grouped in playing positions as: outside backs, centers, halves, loose forwards, lock forwards, and front row forwards. Greater total distance, low-intensity activity, maximal speed, and meters per minute were apparent in matches compared with training in all positions (P < .02; d > 0.90). Similarly, match heart rate and session ratings of perceived exertion responses were higher than those recorded in training (P < .05; d > 0.8). Key skill completions for forwards (ie, scrums, rucks, and lineouts) and backs (ie, kicks) were greater under match conditions than in training (P < .001; d > 1.50). Considerable disparities exist between the perceptual, physiological, and key skill demands of competitive matches versus training sessions in preprofessional rugby union players. Practitioners should consider the specificity of training tasks for preprofessional rugby players to ensure the best preparation for match demands.


Newcastle-Ottawa Scale: comparing reviewers' to authors' assessments.
Lack of appropriate reporting of methodological details has previously been shown to distort risk of bias assessments in randomized controlled trials. The same might be true for observational studies. The goal of this study was to compare the Newcastle-Ottawa Scale (NOS) assessment for risk of bias between reviewers and authors of cohort studies included in a published systematic review on risk factors for severe outcomes in patients infected with influenza. Cohort studies included in the systematic review and published between 2008-2011 were included. The corresponding or first authors completed a survey covering all NOS items. Results were compared with the NOS assessment applied by reviewers of the systematic review. Inter-rater reliability was calculated using kappa (K) statistics. Authors of 65/182 (36%) studies completed the survey. The overall NOS score was significantly higher (p < 0.001) in the reviewers' assessment (median = 6; interquartile range [IQR] 6-6) compared with those by authors (median = 5, IQR 4-6). Inter-rater reliability by item ranged from slight (K = 0.15, 95% confidence interval [CI] = -0.19, 0.48) to poor (K = -0.06, 95% CI = -0.22, 0.10). Reliability for the overall score was poor (K = -0.004, 95% CI = -0.11, 0.11). Differences in assessment and low agreement between reviewers and authors suggest the need to contact authors for information not published in studies when applying the NOS in systematic reviews.


Fourier domain preconditioned conjugate gradient algorithm for atmospheric tomography.
By 'atmospheric tomography' we mean the estimation of a layered atmospheric turbulence profile from measurements of the pupil-plane phase (or phase gradients) corresponding to several different guide star directions. We introduce what we believe to be a new Fourier domain preconditioned conjugate gradient (FD-PCG) algorithm for atmospheric tomography, and we compare its performance against an existing multigrid preconditioned conjugate gradient (MG-PCG) approach. Numerical results indicate that on conventional serial computers, FD-PCG is as accurate and robust as MG-PCG, but it is from one to two orders of magnitude faster for atmospheric tomography on 30 m class telescopes. Simulations are carried out for both natural guide stars and for a combination of finite-altitude laser guide stars and natural guide stars to resolve tip-tilt uncertainty.


Targeting the HIV-1 Spike and Coreceptor with Bi- and Trispecific Antibodies for Single-Component Broad Inhibition of Entry.
Protection against acquiring human immunodeficiency virus (HIV) infection may not require a vaccine in the conventional sense, because broadly neutralizing antibodies (bNAbs) alone prevent HIV infection in relevant animal challenge models. Additionally, bNAbs as therapeutics can effectively suppress HIV replication in infected humans and in animal models. Combinations of bNAbs are generally even more effective, and bNAb-derived multivalent antibody-like molecules also inhibit HIV replication both in vitro and in vivo To expand the available array of multispecific HIV inhibitors, we designed single-component molecules that incorporate two (bispecific) or three (trispecific) bNAbs that recognize HIV Env exclusively, a bispecific CrossMAb targeting two epitopes on the major HIV coreceptor, CCR5, and bi- and trispecifics that cross-target both Env and CCR5. These newly designed molecules displayed exceptional breadth, neutralizing 98 to 100% of a 109-virus panel, as well as additivity and potency compared to those of the individual parental control IgGs. The bispecific molecules, designed as tandem single-chain variable fragments (scFvs) (10E8fv-N6fv and m36.4-PRO 140fv), displayed median 50% inhibitory concentration (IC50s) of 0.0685 and 0.0131 μg/ml, respectively. A trispecific containing 10E8-PGT121-PGDM1400 Env-specific binding sites was equally potent (median IC50 of 0.0135 μg/ml), while a trispecific molecule targeting Env and CCR5 simultaneously (10E8Fab-PGDM1400fv-PRO 140fv) demonstrated even greater potency, with a median IC50 of 0.007 μg/ml. By design, some of these molecules lacked Fc-mediated effector function; therefore, we also constructed a trispecific prototype possessing reconstituted CH2-CH3 domains to restore Fc receptor binding capacity. The molecules developed here, along with those described previously, possess promise as prophylactic and therapeutic agents against HIV.IMPORTANCE Broadly neutralizing antibodies (bNAbs) prevent HIV infection in monkey challenge models and suppress HIV replication in infected humans. Combinations of bNAbs are more effective at suppression, and antibody-like molecules engineered to have two or three bNAb combining sites also inhibit HIV replication in monkeys and other animal models. To expand the available array of multispecific HIV inhibitors, we designed single-component molecules that incorporate two (bispecific) or three (trispecific) bNAb binding sites that recognize the HIV envelope glycoprotein (Env) or the HIV coreceptor (CCR5) or that cross-target both Env and CCR5. Several of the bi- and trispecific molecules neutralized most viruses in a diverse cross-clade panel, with greater breadth and potency than those of the individual parental bNAbs. The molecules described here provide additional options for preventing or suppressing HIV infection.


Characterization of the DNA-binding properties and the transactivation activity of Schistosoma mansoni nuclear receptor fushi tarazu-factor 1alpha (SmFTZ-F1alpha).
A FTZ-F1-related orphan nuclear receptor SmFTZ-F1alpha was previously identified from Schistosoma mansoni. The deduced SmFTZ-F1alpha protein contains a highly conserved DNA binding domain (DBD, C domain), a less conserved ligand binding domain (LBD, E domain) and three highly variable regions, the N-terminal A/B domain (108 aa), a large hinge region (D domain, 1027 aa) and an F domain (220 aa). Herein, we characterize the DNA binding properties and the transactivation activity of SmFTZ-F1alpha. In in vitro assays, SmFTZ-F1alpha bound as a monomer to a response element (FF1RE: TCAAGGTCA) recognized by mammalian steroidogenic factor 1 (SF-1), and to related sequences (p14: TTAAGGTCA and SmFF1a-2: CGAAGGTCA) derived from known schistosome gene promoters. Competition assays with p14 oligonucleotides containing a single mutation at each nucleotide position defined the optimum DNA sequence required for SmFTZ-F1alpha binding. The optimal consensus sequence for SmFTZ-F1alpha binding is TN(A/G)AGGTC(A/G) (N: any base). This sequence is similar but not identical to the SF-1 response element (SFRE) consensus sequence [(T/C)CAAGG(T/C)C(A/G)]. By performing yeast one-hybrid assays, the ability of SmFTZ-F1alpha to bind productively to a p14-derived 9-base pair sequence was demonstrated in vivo. The ability of the full-length SmFTZ-F1alpha to transactivate reporter gene expression was shown to be A/B domain-dependent in a yeast system. In addition, the hinge region contained an unexpected activation function (AF) domain, termed AF-3, while no transactivation activity was detected within the E/F domain. This AF-3 region (from aa 982 to aa 1110) revealed a strong autonomous transactivation activity, which was masked when it was present in the full-length SmFTZ-F1alpha. Taken together, our results suggest that SmFTZ-F1alpha possesses the characteristic DNA binding specificity of FTZ-F1 subfamily members and the capacity to transactivate a reporter gene.


A randomized trial comparing intravitreal triamcinolone and focal/grid photocoagulation for diabetic macular edema: baseline features.
To compare baseline demographic, systemic, and ocular characteristics within age and racial subgroups among participants in this Diabetic Retinopathy Clinical Research Network clinical trial and to compare this cohort with other cohorts enrolled in phase 3 clinical trials for diabetic retinopathy. Thirty-six month, randomized, controlled, multicenter clinical trial of 693 participants with diabetic macular edema enrolled at 88 clinical sites in the United States. Participants were categorized into self-reported race/ethnicity subgroups and into one of three age groups: 18 to <60, 60 to <70, and 70 and older. Mean age of participants was 63 years, 72% were white, and median visual acuity letter score was 62 (approximately 20/63). No substantial difference was identified between racial subgroups for any baseline variable. Older participants were more likely to have Type 2 diabetes mellitus and longer duration disease. The most frequent levels of diabetic retinopathy among 840 study eyes were moderate (level 43) to moderately severe (level 47) nonproliferative disease. While the racial composition of this cohort does not differ from other cohorts in large phase 3 trials that have evaluated participants with diabetic retinopathy, the inclusion of many subjects over age 70 and a better level of glycemic control are distinguishing features.


Factors influencing outcome of surgery for stage I rectal cancer.
Stage I rectal cancer (T1N0M0, T2N0M0) carries excellent prognosis with up to 90% of long term survival rates and complete cure can be expected with curative surgery. However 10 to 15% of these patients show local recurrence and mortality seen in five years. The aim of this study is to analyze the prognostic factors that influence the overall and disease free survival in patients with stage I rectal cancer. Eighty-five patients with stage I (T1N0M0, T2N0M0) rectal cancer that had potentially curative surgery were entered into the study. The patients were evaluated according to age, sex, preoperative carcinoembryonic antigen (CEA) level, type of surgical procedure, tumor distance from anal verge, tumor size, depth of invasion, histological differentiation, presence of tumor ulceration, peritumoral vascular invasion, peritumoral lymphatic invasion and peritumoral perineural invasion. Five year overall and disease free survival rates for the patients were 88% and 74%, respectively. Multivariate analysis showed that independent predictors of recurrence were differentiation and peritumoral vascular invasion, and independent predictor of overall survival was only differentiation. Stage I rectal cancer patients with poor differentiation and peritumoral vascular invasion are at high risk for recurrence and should be evaluated for adjuvant therapies.


Proteome-based analysis of serologically defined tumor-associated antigens in cutaneous lymphoma.
Information on specificities of serological responses against tumor cells in cutaneous lymphoma patients is relatively restricted. To advance the knowledge of serological immune responses against and to assess the scope of tumor antigenicity of cutaneous lymphoma, 1- and 2-dimensional Western blot analyses with sera from patients were combined with proteomics-based protein identification. Testing sera from 87 cutaneous lymphoma patients by 1-dimensional Western blot analysis, 64 cases of seroreactivity against lymphoma cells were found. The positive responses were relatively weak, restricted to few antigens in each case, and heterogeneous. To identify the antigens, proteins of the mycosis fungoides cell line MyLa and primary tumor cells were separated by 2-dimensional gel electrophoresis, Western-blotted and probed with heterogeneous and autologous patient sera. The antigens were identified from silver-stained replica gels by MALDI-TOF mass spectrometry. 14 different antigens were assigned and identified with this proteome-serological approach. Only one, vimentin, had been reported before, the other 13 are new antigens for cutaneous lymphomas.


Collagen peptides modulate the metabolism of extracellular matrix by human dermal fibroblasts derived from sun-protected and sun-exposed body sites.
Clinical data published in recent years have demonstrated positive effects of collagen hydrolysate (CH) on skin aging clinical signs. CH use as food supplement has a long history; however, few studies have addressed the underlying purpose of CH on the cellular and molecular biology of skin cells that could elucidate clinical improvement findings. Wide diversity of characteristics has been reported for dermal fibroblasts derived from different body sites and it is unknown whether collagen peptides could modulate differently cells from chronological aged and photoaged skin areas. This study investigated the influence of CH on the extracellular matrix metabolism and proliferation of human dermal fibroblasts (HDFs) derived from chronological aged (sun-protected) and photoaged (sun-exposed) body sites. CH treatment did not affect cellular proliferation of either cell cultures, but notably modulated cell metabolism in monolayer model, increasing the content of dermal matrix precursor and main protein, procollagen I and collagen I, respectively. These effects were confirmed in the human dermal equivalent model. The increase in collagen content in the cultures was attributed to stimulation of biosynthesis and decreased collagen I metabolism through inhibition of metalloproteinase activity (MMP) 1 and 2. Modulation of CH in dermal metabolism did not differ between cells derived from sun-protected and sun-exposed areas, although lower concentrations of CH seemed to be enough to stimulate sun-exposed-derived HDFs, suggesting more pronounced effect in these cells. This study contributes to understanding the biological effects of CH on skin cells and viability of its use as a functional ingredient in food supplements.


Three hydrogen-rich solutions protect against intestinal injury in uncontrolled hemorrhagic shock.
Intestinal tissue got largely decreased blood supply in uncontrolled hemorrhagic shock, because of limited blood mainly supporting brain, heart, kidney etc. This makes intestine as the primary injury target after uncontrolled hemorrhagic shock. However, limited studies focus on how to protect intestine against hemorrhagic shock. Ringer's solution, pentoxifylline and hypertonic saline are widely used to resuscitate in haemorrhagic shock and sepsis tissue injury. Evidence showed that hydrogen inhibited inflammation and reduced oxidative damage. Here we tested the hypothesis whether hydrogen rich Ringer's, pentoxifyline and hypertonic saline solutions increase the benefit in protecting small intestine from injury in uncontrolled hemorrhagic shock rat model. We tested the anti-inflammation effect of H-Ringer's, HHES and HHSH administration. We found hydrogen-rich solutions treatment groups showed the decreased MDA, MPO, IL-6 and TNF-α levels, and increased SOD, IL-10 comparing with those of non-hydrogen solutions administration groups. Our histological results showed that these three solutions with saturation hydrogen alleviatived the intestinal injury including the intact intestinal villi and less neutrephil infiltration. Our results indicate that these three hydrogen-rich solutions can protect intestinal injure after uncontrolled hemorrhagic shock. The protective effect might be through inhibiting proinflammatory factors, promoting anti-inflammatory cytokines and reducing inflammatory cells infiltration. Our study has potential clinical importance of uncontrolled hemorrhagic shock patient's resuscitation.


Do variations in substitution rates and male mutation bias correlate with life-history traits? A study of 32 mammalian genomes.
Life-history traits vary substantially across species, and have been demonstrated to affect substitution rates. We compute genome-wide, branch-specific estimates of male mutation bias (the ratio of male-to-female mutation rates) across 32 mammalian genomes and study how these vary with life-history traits (generation time, metabolic rate, and sperm competition). We also investigate the influence of life-history traits on substitution rates at unconstrained sites across a wide phylogenetic range. We observe that increased generation time is the strongest predictor of variation in both substitution rates (for which it is a negative predictor) and male mutation bias (for which it is a positive predictor). Although less significant, we also observe that estimates of metabolic rate, reflecting replication-independent DNA damage and repair mechanisms, correlate negatively with autosomal substitution rates, and positively with male mutation bias. Finally, in contrast to expectations, we find no significant correlation between sperm competition and either autosomal substitution rates or male mutation bias. Our results support the important but frequently opposite effects of some, but not all, life-history traits on substitution rates.


Tbx5 and Tbx20 act synergistically to control vertebrate heart morphogenesis.
Members of the T-box family of proteins play a fundamental role in patterning the developing vertebrate heart; however, the precise cellular requirements for any one family member and the mechanism by which individual T-box genes function remains largely unknown. In this study, we have investigated the cellular and molecular relationship between two T-box genes, Tbx5 and Tbx20. We demonstrate that blocking Tbx5 or Tbx20 produces phenotypes that display a high degree of similarity, as judged by overall gross morphology, molecular marker analysis and cardiac physiology, implying that the two genes are required for and have non-redundant functions in early heart development. In addition, we demonstrate that although co-expressed, Tbx5 and Tbx20 are not dependent on the expression of one another, but rather have a synergistic role during early heart development. Consistent with this proposal, we show that TBX5 and TBX20 can physically interact and map the interaction domains, and we show a cellular interaction for the two proteins in cardiac development, thus providing the first evidence for direct interaction between members of the T-box gene family.


Morphological and Molecular Characteristics of the Oak Tree Canker Pathogen, Annulohypoxylon truncatum.
Cankers are localized dead areas in the bark of stems, branches or twigs of many types of trees and shrubs, and are usually caused by fungi. We observed severe canker symptoms in oak trees located in Gyeongnam province in 2011. A total 31 trees were discovered with cankers of varied size, with an average of 48.5 × 15.2 cm. Black, half-rounded globular mound shaped stromata were associated with the cankers, and the asci of the fungi associated with the cankers were cylindrical shaped with their spore-bearing parts being up to 84 µm in length. The average fungal ascospores size was 7.59 × 4.23 µm. The internal transcribed spacer sequence for the canker causing fungus showed 99% similarity to the sequence of Annulohypoxylon truncatum. In this study, the isolated fungus was precisely described and then compared with fungi of similar taxa.


Understanding the heterogeneity and functions of metabolic tissue macrophages.
Growing evidence places tissue-resident macrophages as essential gatekeepers of metabolic organ homeostasis, including the adipose tissue and the pancreatic islets. Therein, macrophages may adopt specific phenotypes and ensure local functions. Recent advances in single cell genomic analyses provide a comprehensive map of adipose tissue macrophage subsets and their potential roles are now better apprehended. Whether they are beneficial or detrimental, macrophages overall contribute to the proper adipose tissue expansion under steady state and during obesity. By contrast, macrophages residing inside pancreatic islets, which may exert fundamental functions to fine tune insulin secretion, have only started to attract attention and their cellular heterogeneity remains to be established. The present review will focus on the latest findings exploring the phenotype and the properties of macrophages in adipose tissue and pancreatic islets, questioning early beliefs and future perspectives in the field of immunometabolism.


Reframing HIV prevention for gay men in the United States.
The HIV epidemic in the United States has affected at least two generations of gay men. Despite numerous efforts to intervene on this public health crisis, HIV infections continue to escalate, especially among young men. This condition is compounded by an ever-growing number of gay men who are aging and living with HIV. We must enact an innovative and proactive vision and framework for HIV prevention that moves us beyond the undertakings rooted in social-cognitive paradigms that have informed this work for the past 25 years. A new framework for HIV prevention must give voice to gay men; must consider the totality of their lives; must delineate the underlying logic, which directs their relation to sex and HIV; and must concurrently respect their diverse life experiences. This approach should be rooted in a biopsychosocial paradigm, should be informed by both theory and practice, and should be directed by three theoretical lenses--a theory of syndemics, developmental theories, and contextual understandings of HIV disease. Taken together, these elements are a call to action for research and practice psychologists who are working to improve the lives of gay men.


Compensation of ring artefacts in synchrotron tomographic images.
Tomographic images are often superimposed by so called ring artefacts. Ring artefacts are concentric rings in the images around the center of rotation of the tomographic setup caused e.g. by differences in the individual pixel response of the detector. They complicate the post processing of the data, i.e. the segmentation of individual image information. Hence, for a quantitative analysis of the tomographic images a significant reduction of these artefacts is essential. In this paper, a simple but efficient method to eliminate such artefacts during the reconstruction is proposed.


AN ASSESSMENT OF THE CONTRACTILE PROPERTIES OF THE SHOULDER MUSCULATURE IN ELITE VOLLEYBALL PLAYERS USING TENSIOMYOGRAPHY.
In volleyball, offensive (Hitters) and defensive players (Non-Hitters) perform differing actions that vary both kinematically and in terms of intensity. This may impose contrasting demands on the musculature involved in performing these actions. Previous research has identified differences in the muscle activation and contractile properties of the lower-body musculature between positions. Additionally, asymmetries between dominant and non-dominant limbs of the upper-body musculature has been observed in athletes performing overhead movements. The aim of this study was to use Tensiomyography (TMG) to examine the contractile properties of the shoulder musculature in elite volleyball players. Cross-sectional study. Thirty-one elite volleyball players participated in this study (Age: 23 ± 2 yrs, Body Mass: 76.5 ± 9.8 kg, Stature: 181 ± 9.3 cm), 26 of which displayed right-limb dominance and five displayed left-limb dominance. Contractile properties of the shoulder musculature including the anterior deltoid (AD), biceps brachii (BB), posterior deltoid (PD), and the upper trapezius (UT) were assessed bilaterally using TMG measures on one occasion prior to any training or exercise. The contractile measures provided by TMG included the maximal displacement (Dm), contraction time (Tc), delay time (Td), sustain time (Ts), and the relaxation time (Tr). No statistically significant differences were observed between positions or limbs, except that Hitters displayed a significantly lower Ts of the left AD compared to Non-hitters (p = 0.01, ES = 1.02), and significant differences between dominant and non-dominant sides in the Td of the UT in Non-hitters were present (p = 0.05, ES = 0.8). These data suggest that irrespective of playing position and limb dominance, contractile properties of the shoulder musculature in elite volleyball players, as measured using TMG, display few significant differences. 3b.


Functional analysis of APC-Cdh1.
To maintain genomic integrity against various kinds of genotoxic stress, cells have multiple checkpoints in the cell cycle. When one of the cell cycle events, such as DNA synthesis, DNA repair, and chromosomal segregation, has not been successfully completed, checkpoints will delay progression until the step is correctly accomplished, and only then will they relieve the arrest to allow the cell to move to the next phase. Cells lacking functional checkpoints display genomic aberrations, resulting in the acquisition of phenotypic changes of cancer cells. Anaphase-promoting complex (APC) is activated by two regulatory proteins: Cdc20 and Cdh1. In yeast and Drosophila, Cdh1-dependent APC (APC-Cdh1) activity targets mitotic cyclins from the end of mitosis to the G1 phase. Loss of Cdh1 induces unscheduled accumulation of mitotic cyclins in G1, resulting in abrogation of G1 arrest caused by treatment with rapamycin, an inducer of p27Kip1. Furthermore, Cdh1-deficient DT40 cells fail to maintain DNA damage-induced G2 arrest, and Cdh1-APC is activated by X-irradiation-induced DNA damage. Thus, activation of Cdh1-APC plays a crucial role in both cdk inhibitor-dependent G1 arrest and DNA damage-induced G2 arrest. In light of the differences between normal and cancer cells, checkpoints can be ideal targets for developmental cancer therapeutics. In this chapter, we describe how to analyze three checkpoints (spindle assembly checkpoint, rapamycin-induced G1 checkpoint, and DNA damage-induced G2 checkpoint) in conjunction with cell synchronization.


Soluble HLA class I antigens in serum of healthy individuals--population study.
Variability of concentrations of s-HLA-I depending on allelic specificity substantiated realisation of research in population of 248 healthy, unrelated individuals. Defined phenotypes from tissue typing of polymorphic HLA complex enabled concentrations measurements of 1553 serum samples for HLA--A, B, and C loci determined antigens. Semi-quantitative technique of inhibition microcytotoxic reaction according to Tait et al. (1981) and Mclean et al. (1983) with usage of policlonal sera anti-HLA was applied. For most of numbers of the allelic specificity the concentration of antigen material in soluble form (s-HLA-I) in blood serum were nominally very high and high. For certain numbers of specificity e.g. HLA--A26, A29, B39, B52, B56, Cw5, Cw6 the percentage of sera, where the s-HLA concentrations were decreased was observed. The results were presented as mean values of inhibition microcytotoxic reaction--according to NIH classification. Authors point on usefulness of results for s-HLA comparative analysis of particular HLA allelic specificity, specific for certain diseases e.g. Cw7 for SNHL patients and B27 for ankylosing spongilitis.


An experience of ultrasound-guided hydrostatic reduction of intrussusception at a tertiary care centre.
Intussusception is an important and one of the most commonly encountered diagnoses of intestinal obstruction in the paediatric age group. Ultrasound-guided hydrostatic reduction is an effective, nonoperative treatment modality for this condition and is associated with a high success rate. In addition, it is simple and safe as the entire procedure is carried out with real-time ultrasound, without the hazard of radiation. The aim of this study was to evaluate the effi cacy and safety of ultrasound-guided hydrostatic reduction in the management of intussusceptions in the paediatric age group. A case study was carried out on 89 patients diagnosed with intussusception using high-resolution ultrasonography over a period of two years, spanning February 2012 to January 2014. Ultrasound-guided hydrostatic reduction was performed in 78 of these patients, and 11 patients were excluded owing to clinical contraindications. Follow-up ultrasound was performed after 24 hours to rule out recurrence. The disease was most prevalent in the age group 6-24 months. The ileocolic type was the most common. Mean duration (hours) was 17.02 ± 20.81 for time to presentation. Complete therapeutic reduction was achieved in 70 of the 78 cases, with a success rate of 90%. Two recurrences occurred in the following 24 hours, which were successfully reduced on the second attempt. Complications and mortality did not occur secondary to the procedure. Our study found that ultrasound-guided hydrostatic reduction is a simple, safe and effective nonoperative treatment for intussusceptions in the paediatric age group, and should be the fi rst line of management in appropriate patients.


E-Cigarettes and Other Electronic Nicotine Delivery Systems (ENDS).
Electronic nicotine delivery systems and vaping devices are now the most commonly used forms of tobacco consumed by youth and young adults. A variety of chemicals and toxicants released during inhalation have harmful systemic effects and effects on the lung. The known and potential health consequences are particularly alarming. There is concern that use of these devices will create a new generation of life-long smokers with nicotine and polysubstance addiction. Beyond the concerning chronic health effects of regular use, there is a recent epidemic of severe, acute lung disease termed e-cigarette, or vaping product use-associated lung injury, (EVALI), associated with high morbidity and mortality. These issues demand immediate public health attention. Healthcare providers play key roles in limiting the exposure of youth and young adults to these products by providing evidence-based counseling to patients and families, and by advocating for regulations to protect against childhood initiation and continuation of use.


Biodistribution in tumour-bearing mice of two 90Y-labelled biotins using three-step tumour targeting.
Tumour pretargeting techniques based on a three-step approach with the avidin/biotin system and associated with the high-energy beta-emitter yttrium-90 (90Y) have been applied with encouraging results in cancer therapy. While in-vivo stable binding of 90Y through a chelating agent like DOTA is essential for directly labelled monoclonal antibodies in the labelling of a fast clearing molecule like biotin, this may not be so important, since the time for catabolic processes to occur is drastically reduced. In this study, the biodistribution of 90Y bound to biotin through DTPA or DOTA was evaluated. Tumour-bearing mice received biotinylated antibody on day 1, avidin on day 2 and 90Y-labelled biotin on day 3. The biodistribution was determined 4 and 24 h after the injection of radiolabelled biotin. The majority of the injected dose was recovered in the urine 4 h after injection of both radiolabelled compounds. Higher bone uptake of 90Y was observed in animals administered 90Y-DTPA-biotin. In general, all organs including tumour showed higher accumulation of activity following the injection of 90Y-DAPA-biotin. Despite the lower levels of accumulation in the tumour after injection of 90Y-DOTA-biotin, the tumour-to-liver and tumour-to-bone ratios for the DOTA ligand were higher compared to DTPA. The higher tumour uptake observed after injection of 90Y-DTPA-biotin may be attributed to the presence of two biotin molecules in its dimeric structure. Our results indicate that the ideal 90Y-labelled biotin derivative for therapy studies should be formed from a DOTA core with two non-cleavable spacers, each ending with a biotin molecule.


Electronic Nicotine Delivery Systems.
Electronic nicotine delivery systems (ENDS) are rapidly growing in popularity among youth. ENDS are handheld devices that produce an aerosolized mixture from a solution typically containing concentrated nicotine, flavoring chemicals, and propylene glycol to be inhaled by the user. ENDS are marketed under a variety of names, most commonly electronic cigarettes and e-cigarettes. In 2014, more youth reported using ENDS than any other tobacco product. ENDS pose health risks to both users and nonusers. Nicotine, the major psychoactive ingredient in ENDS solutions, is both highly addictive and toxic. In addition to nicotine, other toxicants, carcinogens, and metal particles have been detected in solutions and aerosols of ENDS. Nonusers are involuntarily exposed to the emissions of these devices with secondhand and thirdhand aerosol. The concentrated and often flavored nicotine in ENDS solutions poses a poisoning risk for young children. Reports of acute nicotine toxicity from US poison control centers have been increasing, with at least 1 child death reported from unintentional exposure to a nicotine-containing ENDS solution. With flavors, design, and marketing that appeal to youth, ENDS threaten to renormalize and glamorize nicotine and tobacco product use. There is a critical need for ENDS regulation, legislative action, and counter promotion to protect youth. ENDS have the potential to addict a new generation of youth to nicotine and reverse more than 50 years of progress in tobacco control.


Assembly and performance of a 6.4 T cryogen-free dynamic nuclear polarization system.
We report on the assembly and performance evaluation of a 180-GHz/6.4 T dynamic nuclear polarization (DNP) system based on a cryogen-free superconducting magnet. The DNP system utilizes a variable-field superconducting magnet that can be ramped up to 9 T and equipped with cryocoolers that can cool the sample space with the DNP assembly down to 1.8 K via the Joule-Thomson effect. A homebuilt DNP probe insert with top-tuned nuclear magnetic resonance coil and microwave port was incorporated into the sample space in which the effective sample temperature is approximately 1.9 K when a 180-GHz microwave source is on during DNP operation. 13 C DNP of [1-13 C] acetate samples doped with trityl OX063 and 4-oxo-TEMPO in this system have resulted in solid-state 13 C polarization levels of 58 ± 3% and 18 ± 2%, respectively. The relatively high 13 C polarization levels achieved in this work have demonstrated that the use of a cryogen-free superconducting magnet for 13 C DNP is feasible and in fact, relatively efficient-a major leap to offset the high cost of liquid helium consumption in DNP experiments.


Cortisone-induced recrudescence of Giardia lamblia infections in gerbils.
The course of infection with Giardia lamblia in gerbils was studied after oral inoculation with either cysts isolated from humans or cultured trophozoites. Primary infections occurred in 87% of exposed gerbils; cyst release peaked during the second and third weeks post-infection; cysts and trophozoites were observed for up to 49 and 87 days post-infection, respectively. Gerbils were refractory to challenge infection with infective human source cysts at 5 months post-infection. Overall, 50% of animals given subcutaneous injections of hydrocortisone acetate, during or following the late stages of their primary infection, showed recrudescence of infection as evidenced by passage of cysts; percentages showing recrudescence were: 80% at 50 days post-infection, 55% at 70 days post-infection, 27% at 7 months post-infection, and 50% (2 of 4) in gerbils challenged unsuccessfully at 5 months post-infection and subsequently injected with hydrocortisone acetate at 7 months post-infection. Both infected animals and uninfected controls injected with hydrocortisone acetate were immunocompromised as evidenced by significantly reduced plaque forming cell responses to sheep red blood cells. We conclude that infections with G. lamblia in gerbils can last longer than standard examinations for cysts would indicate, and that long-term immunity to this organism can be nonsterile.


First-pass metabolism of gentisamide: influence of intestinal metabolism on hepatic formation of conjugates. Studies in the once-through vascularly perfused rat intestine-liver preparation.
Gentisamide (GAM) metabolism was studied in a once-through in situ perfused rat intestine-liver preparation in a manner which mimicked the first-pass effect. GAM (2.56 to 392 microM) was delivered into the intestine via the superior mesenteric artery at a flow of 7.5 ml/min. The intestinal venous outflow into the portal vein and the hepatic arterial flow (2.5 ml/min; without drug) served as dual inflows into the liver. The steady state intestinal extraction ratios (E1, 0.33 to 0.06) and hepatic extraction ratios (EH, 0.84 to 0.37) were highly concentration dependent, decreasing with increasing GAM concentrations. Gentisamide-5-glucuronide was found to be the major intestinal metabolite, and gentisamide-2-sulfate was also formed, albeit to a much lesser extent. However, hepatic metabolism of GAM revealed 5-sulfation and glucuronidation as the predominant pathways, with GAM 2-sulfation as a minor pathway. About 1 to 3% and 7% of dose was excreted into lumen and bile, respectively, mostly as GAM-5G. From the metabolic data, the fitted parameters confirmed that more enzymes exist in the liver than in intestine for the processing of GAM. The Km for 2-sulfation by the intestine was higher than the apparent Kms observed for 2- and 5-sulfation in liver. However, the Kms for 5-glucuronidation for both the intestine and liver were similar. The interrelationship between intestine and liver formation of first-pass metabolites showed that, at low GAM input concentration entering the IL preparation, the intestine, being the anterior organ, regulated the available substrate for hepatic elimination, and hence reduced the contribution of hepatic metabolism in the overall first-pass effect.(ABSTRACT TRUNCATED AT 250 WORDS)


Factors affecting cadence choice during submaximal cycling and cadence influence on performance.
Cadence choice during cycling has been of considerable interest among cyclists, coaches, and researchers for nearly 100 years. The present review examines and summarizes the current knowledge of factors affecting the freely chosen cadence during submaximal cycling and of the influence of cadence choice on performance. In addition, suggestions for future research are given along with scientifically based, practical recommendations for those involved in cycling. Within the past 10 years, a number of papers have been published that have brought novel insight into the subject. For example, under the influence of spinal central pattern generators, a robust innate voluntary motor rhythm has been suggested as the primary basis for freely chosen cadence in cycling. This might clarify the cadence paradox in which the freely chosen cadence during low-to-moderate submaximal cycling is considerably higher and thereby less economical than the energetically optimal cadence. A number of factors, including age, power output, and road gradient, have been shown to affect the choice of cadence to some extent. During high-intensity cycling, close to the maximal aerobic power output, cyclists choose an energetically economical cadence that is also favorable for performance. In contrast, the choice of a relatively high cadence during cycling at low-to-moderate intensity is uneconomical and could compromise performance during prolonged cycling.


Non-null annular subaperture stitching interferometry for steep aspheric measurement.
A non-null annular subaperture stitching interferometry (NASSI), combining the subaperture stitching ideal and non-null test method, is proposed for steep aspheric testing. Compared with standard annular subaperture stitching interferometry (ASSI), a partial null lens (PNL) is employed as an alternative to the transmission sphere, to generate different aspherical wavefronts as the references. The coverage subaperture number would be reduced greatly for the better performance of aspherical wavefronts in matching the local slope of aspheric surfaces. In this way, relatively large overlapping areas can be obtained for adjustment errors correction while the error accumulation would be decreased. With the reverse optimization reconstruction (ROR) method for retrace error correction, the figure error of each subaperture can be retrieved accurately. Therefore, the testing accuracy and efficiency are thus increased. The dynamic test range is extended as well. A numerical simulation exhibits the comparison of the performance of the NASSI and standard ASSI, which demonstrates the high accuracy of the NASSI in testing steep aspheric. Experimental results of NASSI are shown to be in good agreement with those of the Zygo interferometer.


Epidemiological data and genome sequencing reveals that nosocomial transmission of SARS-CoV-2 is underestimated and mostly mediated by a small number of highly infectious individuals.
Despite robust efforts, patients and staff acquire SARS-CoV-2 infection in hospitals. We investigated whether whole-genome sequencing enhanced the epidemiological investigation of healthcare-associated SARS-CoV-2 acquisition. From 17-November-2020 to 5-January-2021, 803 inpatients and 329 staff were diagnosed with SARS-CoV-2 infection at four Oxfordshire hospitals. We classified cases using epidemiological definitions, looked for a potential source for each nosocomial infection, and evaluated genomic evidence supporting transmission. Using national epidemiological definitions, 109/803(14%) inpatient infections were classified as definite/probable nosocomial, 615(77%) as community-acquired and 79(10%) as indeterminate. There was strong epidemiological evidence to support definite/probable cases as nosocomial. Many indeterminate cases were likely infected in hospital: 53/79(67%) had a prior-negative PCR and 75(95%) contact with a potential source. 89/615(11% of all 803 patients) with apparent community-onset had a recent hospital exposure. Within 764 samples sequenced 607 genomic clusters were identified (>1 SNP distinct). Only 43/607(7%) clusters contained evidence of onward transmission (subsequent cases within ≤ 1 SNP). 20/21 epidemiologically-identified outbreaks contained multiple genomic introductions. Most (80%) nosocomial acquisition occurred in rapid super-spreading events in settings with a mix of COVID-19 and non-COVID-19 patients. Current surveillance definitions underestimate nosocomial acquisition. Most nosocomial transmission occurs from a relatively limited number of highly infectious individuals.


High-resolution electrophysiology on a chip: Transient dynamics of alamethicin channel formation.
Microstructured planar substrates have been shown to be suitable for patch clamp recording from both whole cells and isolated patches of membrane, as well as for measurements from planar lipid bilayers. Here, we further explore this technology with respect to high-resolution, low noise single-channel recording. Using solvent-free lipid bilayers from giant unilamellar vesicles obtained by electro-swelling, we recorded channels formed by the peptaibol alamethicin, a well-studied model system for voltage-dependent channels, focusing on the transient dynamics of single-channel formation upon application of a voltage step. With our setup, we were able to distinctly resolve dwell times well below 100 mus and to perform a thorough statistical analysis of alamethicin gating. Our results show good agreement with models that do not rely on the existence of non-conducting preaggregate states. Microstructured apertures in glass substrates appear promising with respect to future experiments on cellular ion channels reconstituted in suspended lipid membranes.


Nuclear DNA content and p53 overexpression in stage I squamous cell carcinoma of the tongue compared with advanced tongue carcinomas.
To evaluate the predictive value of the nuclear DNA content (image cytometry) and p53 overexpression (immuno-histochemistry using antibody CM-1) in uniformly treated stage I carcinomas of the mobile tongue. Also, to compare stage I carcinomas with advanced tongue carcinomas (stages II-IV). Archival formalin fixed, paraffin wax embedded tumour specimens from 54 patients with stage I squamous cell carcinoma and 37 patients with advanced squamous cell carcinoma were analysed. Mean follow up time of the stage I carcinomas was 71 months (median, 62.5; range, 6-175). Twenty three patients (stage I) had recurring disease: 10 had local recurrence (in the tongue) and 13 had regional recurrence (cervical metastases). Locally recurring stage I carcinomas had a more pronounced DNA deviation than the other stage I carcinomas and this degree of deviation was comparable with the DNA content of advanced carcinomas. Stage I carcinomas that developed regional recurrences overexpressed p53 more frequently. In Cox multivariate regression analysis of time to recurrence, DNA deviation was a significant parameter in tumours that recurred locally (p = 0.032). p53 overexpression was the only parameter close to significance for regional recurrence (p = 0.065). Nuclear DNA content and p53 immunostaining are of value for the prediction of recurrence of stage I squamous cell carcinomas of the mobile tongue. Stage I tongue carcinomas that are prone to local recurrence show the same DNA content as do advanced tongue carcinomas.


Directional asymmetry in the volume of the human habenula.
Brain asymmetry is a conserved feature in vertebrates. The dorsal diencephalic habenular complex shows conspicuous structural and functional asymmetries in a wide range of species, yet it is unclear if this condition is also present in humans. Addressing this possibility becomes relevant in light of recent findings presenting the habenula as a novel target for therapeutic intervention of affective disorders through deep brain stimulation. Here we performed volumetric analyses in postmortem diencephalic samples of male and female individuals, and report for the first time, the presence of directional asymmetries in the volume of the human habenula. The habenular volume is larger on the left side in both genders, a feature that can be explained by an enlargement of the left lateral habenula compared to the right counterpart. In contrast, the volume of the medial habenula shows no left-right directional bias in either gender. It is remarkable that asymmetries involve the lateral habenula, which in humans is particularly enlarged compared to other vertebrates and plays relevant roles in aversive processing and aversively motivated learning. Our findings of structural asymmetries in the human habenula are consistent with recent observations of lateral bias in activation, metabolism and damage of the human habenula, highlighting a potential role of habenular laterality in contexts of health and illness.


Neighborhood disparities in stroke and myocardial infarction mortality: a GIS and spatial scan statistics approach.
Stroke and myocardial infarction (MI) are serious public health burdens in the US. These burdens vary by geographic location with the highest mortality risks reported in the southeastern US. While these disparities have been investigated at state and county levels, little is known regarding disparities in risk at lower levels of geography, such as neighborhoods. Therefore, the objective of this study was to investigate spatial patterns of stroke and MI mortality risks in the East Tennessee Appalachian Region so as to identify neighborhoods with the highest risks. Stroke and MI mortality data for the period 1999-2007, obtained free of charge upon request from the Tennessee Department of Health, were aggregated to the census tract (neighborhood) level. Mortality risks were age-standardized by the direct method. To adjust for spatial autocorrelation, population heterogeneity, and variance instability, standardized risks were smoothed using Spatial Empirical Bayesian technique. Spatial clusters of high risks were identified using spatial scan statistics, with a discrete Poisson model adjusted for age and using a 5% scanning window. Significance testing was performed using 999 Monte Carlo permutations. Logistic models were used to investigate neighborhood level socioeconomic and demographic predictors of the identified spatial clusters. There were 3,824 stroke deaths and 5,018 MI deaths. Neighborhoods with significantly high mortality risks were identified. Annual stroke mortality risks ranged from 0 to 182 per 100,000 population (median: 55.6), while annual MI mortality risks ranged from 0 to 243 per 100,000 population (median: 65.5). Stroke and MI mortality risks exceeded the state risks of 67.5 and 85.5 in 28% and 32% of the neighborhoods, respectively. Six and ten significant (p < 0.001) spatial clusters of high risk of stroke and MI mortality were identified, respectively. Neighborhoods belonging to high risk clusters of stroke and MI mortality tended to have high proportions of the population with low education attainment. These methods for identifying disparities in mortality risks across neighborhoods are useful for identifying high risk communities and for guiding population health programs aimed at addressing health disparities and improving population health.


Modeling and experimental verification of optical materials formed by stacked nanostrips.
The effective plasma frequency fp of periodic metallic wires whose characteristic dimensions are comparable to their skin depth has been analyzed. And a relevant analytic model is constructed by considering the skin effect and making a reasonable shape approximation, which is suitable for the case that the cross section of the wire is noncircular. To verify this model, a wires array with rectangle cross section is designed and the corresponding stacked Au-SiO(2) nanostrips are fabricated. The experimental and simulational transmittances of the metamaterial have been evaluated with a good agreement, although the presence of quartz substrate and structural imperfections in experiment will have an impact, which validates that the multilayer Au-SiO(2) nanostrips could function similarly to a natural bulk metal with discrepancies of fp values less than 8%. It could be confirmed that the theoretic formula is trustworthy in predicting fp for designing and realizing a controllable artificial metal in optical region.


Circular economy through waste reverse logistics under extended producer responsibility in Finland.
Extended producer responsibility (EPR) is commonly implemented as a strategy in waste management. The core of the concept itself is a waste reverse logistics (WRL), which dictates how the collection, inspection and processing of end-of-life products are performed. Existing studies of EPR mainly focused on single products instead of using broader perspective on national level. Its contribution towards circular economy through slowing and closing the loops also has not been widely discussed. This study examined the system architecture of the policy instruments used in the EPR and the similarities of the WRL networks across different products. A case study was used to investigate six products: portable batteries and accumulators, paper, packaging, vehicles, electrical and electronic equipment (EEE) and tyres. The study generated a WRL framework. It is also observed that closing the loop through recycling is the primary circular strategy and is found in all products, whereas closing and slowing the loop strategy through reuse/repair, remanufacture and repurposing is found in packaging, tyres, vehicles and EEE. This study shows that EPR can close the material loop, although improvement in design for the environment is necessary. It creates challenges and opportunities for the government, producer responsibility organization and producers to improve existing conditions by implementing new initiatives such as design for the environment indicators, standardization, tax and subsidy systems and tariffs for disposal fees.


Kinetic Basis for DNA Target Specificity of CRISPR-Cas12a.
Class 2 CRISPR-Cas nucleases are programmable genome editing tools with promising applications in human health and disease. However, DNA cleavage at off-target sites that resemble the target sequence is a pervasive problem that remains poorly understood mechanistically. Here, we use quantitative kinetics to dissect the reaction steps of DNA targeting by Acidaminococcus sp Cas12a (also known as Cpf1). We show that Cas12a binds DNA tightly in two kinetically separable steps. Protospacer-adjacent motif (PAM) recognition is followed by rate-limiting R-loop propagation, leading to inevitable DNA cleavage of both strands. Despite functionally irreversible binding, Cas12a discriminates strongly against mismatches along most of the DNA target sequence. This result implies substantial reversibility during R-loop formation-a late transition state-and defies common descriptions of a "seed" region. Our results provide a quantitative basis for the DNA cleavage patterns measured in vivo and observations of greater reported target specificity for Cas12a than for the Cas9 nuclease.