An inexpensive device for freeze drying and plastic embedding tissues at low temperatures.
The preparation of biological tissues for electron microscopy by rapid freezing retains the original localization of ions and molecules. A reproducible freezing regime was established by quenching tissues in liquid propane according to the method of Elder et al. (1981). Tissue was thereafter freeze dried in a custom built freeze drying device with a liquid nitrogen cooled stage to prevent ice recrystallization during drying. The device was also designed to allow the vacuum embedding of tissue in low temperature resin such as Lowicryl and polymerization in situ. This paper describes the design of the device and an example of its use in the freeze drying of cartilage. The results show that minimal ice damage occurs to the chondrocytes and that intracellular organelles are clearly visible. The regime described may prove a useful and pragmatic alternative to cutting tissue in the frozen state. Translocation of elements is unlikely except perhaps in the case of very labile elements such as Na and K, but this remains to be fully elucidated.


Toxicokinetics of 2,4- and 2,6-toluenediamine in hydrolysed urine and plasma after occupational exposure to 2,4- and 2,6- toluene diisocyanate.
To assess the toxicokinetics of 2,4- and 2,6- toluenediisocyanate (TDI) in chronically exposed subjects. Blood and urine, from 11 workers at two flexible foam polyurethane production plants, were sampled. By gas chromatography-mass spectrometry (GC-MS) 2,4- and 2,6-toluene diamine (TDA) were measured as pentafluoropropionic anhydride (PFPA) derivatives after acidic hydrolysis of plasma (P-TDA, ng/ml) and urine (U-TDA, microgram/h). In one of the plants the P-2,4-TDA concentrations were 0.4-1 ng/ml before a four to five week holiday and 0.2-0.5 ng/ml afterwards. The corresponding values for P-2,6-TDA were 2-6 and 0.5-2 ng/ml respectively. In the other plant the P-2,4-TDA concentrations were 2-23 ng/ml before the holiday and 0.5-6 ng/ml afterwards and the P-2,6-TDA concentrations were 7-24 ng/ml before and 3-6 ng/ml afterwards. The P-2,4-TDA concentrations were 2-24 ng/ml before a 12 day holiday, and 1-14 ng/ml afterwards. The corresponding values for P-2,6-TDA were 12-29 and 8-17 ng/ml, respectively. The urinary elimination rates (U-TDA, microgram/h) for 2,4-TDA before the holiday were 0.04-0.54 and 0.02-0.18 microgram/h afterwards. The corresponding values for 2,6-TDA were 0.18-0.76 microgram/h before and 0.09-0.27 microgram/h after the holiday. The half life in urine ranged between 5.8 and 11 days for 2,4- and 2,6-TDA. The differences in exposure were reflected by the P-TDA concentrations. The mean half life in plasma was 21 (range 14-34) days for 2,4-TDA and 21 (16-26) days for 2,6-TDA. The TDI air concentrations varied between 0.4 and 4 micrograms/m3 in one plant and in the other between 10 and 120 micrograms/m3. The half life in plasma of chronically exposed workers for 2,4-and 2,6-TDA was twice as long as for volunteers with short term exposure. An indication of a two phase elimination pattern in urine was found. The first phase was related to the more recent exposure and the second, much slower one was probably related to release of TDA in urine from TDI adducts in the body.


A case series of rare neurological and cardio-pulmonary manifestations of thiamine deficiency in pregnancy and lactation.
Thiamine (vitamin B1) is a sulfur-containing, water-soluble vitamin that plays an essential role in energy metabolism and the tricarboxylic acid cycle. There is an increased need for vitamin B1 (1-1.2 mg daily) during pregnancy and lactation. Hyperemesis during pregnancy can cause severe polyneuropathy (Wernicke's encephalopathy) as a result of thiamine deficiency. Thiamine deficiency has also been associated with beriberi. A number of atypical cases with reversible right heart failure and severe pulmonary hypertension have also been reported in non-pregnant individuals, but have never been reported in pregnancy. Here we present five such cases of thiamine deficiency with neurological and cardio-pulmonary manifestations.


Role of tobacco warning labels in informing smokers about risks of smoking among bus drivers in Mangalore, India.
Smoking tobacco is considered as a leading cause of preventable death, mostly in developing countries like India. One of the primary goals of international tobacco control is to educate smokers about the risks associated with tobacco consumption. Tobacco warning labels (TWLs) on cigarette packages are one of the most common statutory means to communicate health risks of smoking to smokers, with the hope that once educated, they will be more likely to quit the habit. The present survey was conducted to assess the effectiveness of TWLs in communicating health risks of tobacco usage among 263 adult smokers working as bus drivers in Karnataka State Road Transport Corporation (KSRTC), Mangalore, India. Information was collected on demographic details, exposure and response to health warnings on tobacco products, intention to quit and nicotine dependency. The majority (79.5%) of the respondents revealed negative intentions towards quitting smoking. Nearly half of the participants had a 'low' nicotine dependency (47.5%) and 98.1% of the respondents had often noticed warning labels on tobacco packages. These health warnings made 71.5% of the respondents think about quitting smoking. Respondents who noticed advertisement or pictures about dangers of smoking had better knowledge, with respect to lung cancer and impotence as a consequence of tobacco. A higher exposure to warning labels was significantly associated with lower nicotine dependency levels of smokers among the present study population. A significantly higher number of respondents who noticed advertisement or pictures about the dangers of smoking thought about the risks of smoking and were more inclined to think about quitting smoking. As exposure increased, an increase in the knowledge and response of participants was also observed. Exposure to tobacco warning labels helps to educate smokers about health risks of tobacco smoking. It may be possible to promote oral health among bus drivers by developing strategies to educate them about these risk factors.


Dynamics of virus-specific T cell immunity in pediatric liver transplant recipients.
Immunosuppression following solid organ transplantation (SOT) has a deleterious effect on cellular immunity leading to frequent and prolonged viral infections. To better understand the relationship between posttransplant immunosuppression and circulating virus-specific T cells, we prospectively monitored the frequency and function of T cells directed to a range of latent (CMV, EBV, HHV6, BK) and lytic (AdV) viruses in 16 children undergoing liver transplantation for up to 1 year posttransplant. Following transplant, there was an immediate decline in circulating virus-specific T cells, which recovered posttransplant, coincident with the introduction and subsequent routine tapering of immunosuppression. Furthermore, 12 of 14 infections/reactivations that occurred posttransplant were successfully controlled with immunosuppression reduction (and/or antiviral use) and in all cases we detected a temporal increase in the circulating frequency of virus-specific T cells directed against the infecting virus, which was absent in 2 cases where infections remained uncontrolled by the end of follow-up. Our study illustrates the dynamic changes in virus-specific T cells that occur in children following liver transplantation, driven both by active viral replication and modulation of immunosuppression.


SIMON, an Automated Machine Learning System, Reveals Immune Signatures of Influenza Vaccine Responses.
Machine learning holds considerable promise for understanding complex biological processes such as vaccine responses. Capturing interindividual variability is essential to increase the statistical power necessary for building more accurate predictive models. However, available approaches have difficulty coping with incomplete datasets which is often the case when combining studies. Additionally, there are hundreds of algorithms available and no simple way to find the optimal one. In this study, we developed Sequential Iterative Modeling "OverNight" (SIMON), an automated machine learning system that compares results from 128 different algorithms and is particularly suitable for datasets containing many missing values. We applied SIMON to data from five clinical studies of seasonal influenza vaccination. The results reveal previously unrecognized CD4+ and CD8+ T cell subsets strongly associated with a robust Ab response to influenza Ags. These results demonstrate that SIMON can greatly speed up the choice of analysis modalities. Hence, it is a highly useful approach for data-driven hypothesis generation from disparate clinical datasets. Our strategy could be used to gain biological insight from ever-expanding heterogeneous datasets that are publicly available.


Analysing teamwork in health care: What matters when clinicians negotiate the continuity of clinical tasks and care responsibilities?
This paper considers the ways in which clinicians enact 'being a team', by analysing how they inform one another about critical patient information. The process where this information exchange happens is known as 'clinical handover'. The study that informs this paper spanned ten months of data collection in four hospitals, involving 150 clinicians and five patients. The analysis presented here draws on data collected at one of the hospital sites: an emergency department at a regional tertiary teaching hospital. Our analysis reveals how central clinical handover is to 'being a team' in health care, and how deficiencies in handover weaken clinical teams' ability to provide continuous and safe care for their patients. We further discuss how clinical practitioners' own responses to the footage foregrounded different issues compared to those revealed by formal analysis; namely, issues centring on relationships, and on practical steps to ensure these relationships could be improved. We reflect on the differences between our own formal analysis of the footage, and practitioners' 'lived response' to the footage, and the implications of these differences for how we as analysts conceptualise 'teamness' in organisations.


Family history of breast cancer in relation to tumor characteristics and mortality in a population-based study of young women with invasive breast cancer.
Inherited predisposition may be associated with distinctive breast cancer phenotypes and/or mortality. Past studies have had inconsistent results and little is known about the contributions of screening and treatment. Within a population-based cohort of 1,260 women diagnosed with invasive breast cancer before age 46, we assessed how family history of breast cancer relates to mortality and tumor characteristics. Analyses were repeated excluding BRCA1/BRCA2 carriers. Medical records were reviewed for treatment history and tumors were centrally reviewed and tested. Cox proportional hazard modeling was used to assess the risk of dying in relation to family history; logistic regression was used to assess the association of family history to tumor characteristics. Compared with women with no family history, women with first-degree family history of breast cancer had a 40% reduction (95% CI: 0.5-0.8) in the risk of dying. Mortality in women with only a second-degree family history was similar to those with no family history. The risk of dying was further reduced in those with a greater number of affected relatives. These relationships did not seem to be attributable to differences in screening, detection method, or treatment. Tumors in women with a first-degree family history had generally more favorable prognostic profiles. Our findings suggest that breast cancer patients with a first-degree family history, compared with their counterparts without such a profile, may have a better prognosis. These findings support the need for future research directed at replicating these results and identifying factors underlying this possible relationship.


Acupuncture for spasticity after stroke: a systematic review and meta-analysis of randomized controlled trials.
The aim of this systematic review was to determine how effective acupuncture or electroacupuncture (acupuncture with electrical stimulation) is in treating poststroke patients with spasticity. We searched publications in Medline, EMBASE, and the Cochrane Library in English, 19 accredited journals in Korean, and the China Integrated Knowledge Resources Database in Chinese through to July 30, 2013. We included randomized controlled trials (RCTs) with no language restrictions that compared the effects of acupuncture or electroacupuncture with usual care or placebo acupuncture. The two investigators assessed the risk of bias and statistical analyses were performed. Three RCTs in English, 1 in Korean, and 1 in Chinese were included. Assessments were performed primarily with the Modified Ashworth Scale (MAS). Meta-analysis showed that acupuncture or electroacupuncture significantly decreased spasticity after stroke. A subgroup analysis showed that acupuncture significantly decreased wrist, knee, and elbow spasticity in poststroke patients. Heterogeneity could be explained by the differences in control, acupoints, and the duration after stroke occurrence. In conclusion, acupuncture could be effective in decreasing spasticity after stroke, but long-term studies are needed to determine the longevity of treatment effects.


Decision Aid Implementation among Left Ventricular Assist Device Programs Participating in the DECIDE-LVAD Stepped-Wedge Trial.
Background. Despite demonstrated efficacy, patient decision aids (DAs) are rarely used in clinical practice in the absence of coverage mandates. Deciding whether to pursue a left ventricular assist device (LVAD) is a major, preference-sensitive decision-ideal for exploring implementation of a DA. Methods. We conducted a type II effectiveness-implementation hybrid trial at 6 LVAD programs using a stepped-wedge cluster-randomized design. Using the RE-AIM framework, we collected both quantitative and qualitative outcomes, including a checklist collected by study staff for each enrolled patient regarding DA use and interviews with LVAD program clinicians preintervention, 6 months postintervention, and at the conclusion of the study. Results. From June 2015 to January 2017, 248 patients and their caregivers were enrolled. A total of 69 interviews were conducted with 48 clinicians at 3 time points. The DA reached 95% of eligible patients. Adoption was 100%, as all sites approached agreed to participate in the trial. Interviews revealed several themes related to the implementation of the DA: clinicians had a strong desire to ensure patients were informed and embraced the DA. Despite this, they reported communication challenges among their team that impeded implementation. Five of the 6 sites have maintained use of the DA following the trial; 1 site reported concerns about decreased procedural volume with use of the DA as a reason for discontinuation. Conclusions. In this hybrid trial, a DA for patients considering LVADs and their caregivers demonstrated high reach. Adoption and implementation were facilitated by a strong desire to ensure that patients were well informed. Future dissemination research and practice should attend to concerns about procedure volume and coverage mandates and facilitate ongoing communication at sites using the DA.


Tumor microenvironment in focus: LA-ICP-MS bioimaging of a preclinical tumor model upon treatment with platinum(IV)-based anticancer agents.
The selection of drug candidates for entering clinical development relies on in vivo testing in (solid) tumor animal models. However, the heterogeneity of tumor tissue (e.g. in terms of drug uptake or tissue composition) is rarely considered when testing novel drug candidates. Therefore, we used the murine colon cancer CT-26 tumor model to study the spatially-resolved drug distribution in tumor tissue upon repetitive treatment of animals over two weeks with three investigational platinum(IV)-based anticancer agents, oxaliplatin or satraplatin. A quantitative laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) imaging method revealed a heterogeneous platinum distribution, which correlated well with the histologic features of the tumor and surrounding tissue at the microscopic level. In most of the cases, higher amounts of intratumoral platinum were found in the surrounding tissue than in the malignant parts of the sample. This indicates that determination of average platinum amounts (e.g. by microwave-assisted digestion of the sample followed by analysis with ICP-MS) might overestimate the drug uptake in tumor tissue causing misleading conclusions. In addition, we studied the platinum distribution in the kidneys of treated animals to probe if accumulation in the cortex and medulla predict potential nephrotoxicity. A 10-fold increase of platinum in the cortex of the kidney over the medulla was observed for oxaliplatin and satraplatin. Although these findings are similar to those in the platinum distribution of the nephrotoxic anticancer drug cisplatin, treatment with the compounds of our study did not show signs of nephrotoxicity in clinical use or clinical trials (oxaliplatin, satraplatin) and did not result in the alteration of renal structures. Thus, predicting the side effects based on bioimaging data by LA-ICP-MS should be considered with caution. To the best of our knowledge, this is the first LA-ICP-MS study on spatially-resolved platinum accumulation in tissues after repetitive platinum-based anticancer drug treatment of mice bearing a preclinical tumor model.


Correlation between cervical HPV DNA detection and HPV16 seroreactivity measured with L1-only and L1+L2 viral capsid antigens.
Introduction. Persistent human papillomavirus (HPV) type 16 infection is the main causal agent of cervical cancer. Most HPV infections clear spontaneously within 1-2 years. Although not all infected women develop detectable HPV antibodies, about 60-70 % seroconvert and retain their antibodies at low levels.Aim. We investigated if cervical HPV16 DNA positivity was associated with HPV16 seroreactivity measured with two different antigen formulations. We assessed if associations were influenced by co-infection with other HPV types and HPV16 viral load.Methodology. We used baseline data for women participating in the Ludwig-McGill cohort, a longitudinal investigation of the natural history of HPV infection and cervical neoplasia. The study enrolled 2462 Brazilian women from 1993 to 1997 (pre-vaccination). ELISA assays were based on L1-only or L1+L2 virus-like particles (VLPs). Seroreactivity was expressed as normalized absorbance ratios. HPV genotyping and viral load were evaluated by PCR protocols. Pearson's r was used to measure correlations between interval-scaled variables. Serological accuracy in HPV16 DNA detection was assessed using receiver operating characteristic (ROC) curves. We analysed the association between HPV DNA positivity and HPV16 seroreactivity by linear regression.Results. Correlations between L1+L2 and L1-only VLPs for detection of HPV16 were poor (r=0.43 and 0.44 for dilutions 1 : 10 and 1 : 50, respectively). The protocol with the best accuracy was L1+L2 VLPs at serum dilution 1 : 10 (ROC area=0.73, 95 % CI: 0.65-0.85). HPV16 DNA positivity was correlated with HPV16 seroreactivity and was not influenced by co-infection or viral load. To a lesser degree, HPV16 seroreactivity was correlated with infection by other Alpha-9 papillomavirus species.Conclusion. HPV16 DNA positivity and HPV16 seroreactivity are strongly correlated. L1+L2 VLPs perform better than L1-only VLPs for detecting IgG antibodies to HPV16 in women infected with HPV16 or other Alpha-9 HPV species. This study advances our understanding of humoral immune responses against HPV16 by providing insights about the influence of VLP antigen composition to measure humoral immune response against naturally acquired HPV infection.


[Declared diseases, observed diseases and health priorities in a Benin rural district].
A project of a humanitarian action was preceded by a health diagnosis of a population in the rural district Bonou in Bénin. A diagnosis of perception was set up by the interviews among the population and the local health professionals. That diagnosis was completed by an objectified diagnosis originating from the analysis of 460 consultations performed in Bénin end 1998- beginning 1999. The cross-sorting between the diagnosis of perception and the objectified diagnosis pointed out four public health problems: malaria, bronchial asthma, infectious diarrhoeas and Buruli ulcer with respective prevalences of 9%, 6%, 3% and 0.9%. The prevalence of bronchial asthma fluctuates from 4% in dry areas to 8% in wet areas (Chi 2 = 3.50; p = 0.06). The role of house dust mites is suspected because of those ecological arguments to which is added clinical reasoning. Bronchial asthma was chosen as a priority health problem because of the feasibility and acceptance of a survey which aims at pointing out the allergic etiology. The confirmation of that etiology will enable in that case the decrease of the allergenic pressure, particularly well adapted solution in the Southern countries. Therefore a humanitarian action is foreseen to apprehend the etiologies of bronchial asthma in the rural district Bonou in Bénin in 2000.


Effect of anakinra on mortality in patients with COVID-19: a systematic review and patient-level meta-analysis.
Anakinra might improve the prognosis of patients with moderate to severe COVID-19 (ie, patients requiring oxygen supplementation but not yet receiving organ support). We aimed to assess the effect of anakinra treatment on mortality in patients admitted to hospital with COVID-19. For this systematic review and individual patient-level meta-analysis, a systematic literature search was done on Dec 28, 2020, in Medline (PubMed), Cochrane, medRxiv, bioRxiv, and the ClinicalTrials.gov databases for randomised trials, comparative studies, and observational studies of patients admitted to hospital with COVID-19, comparing administration of anakinra with standard of care, or placebo, or both. The search was repeated on Jan 22, 2021. Individual patient-level data were requested from investigators and corresponding authors of eligible studies; if individual patient-level data were not available, published data were extracted from the original reports. The primary endpoint was mortality after 28 days and the secondary endpoint was safety (eg, the risk of secondary infections). This study is registered on PROSPERO (CRD42020221491). 209 articles were identified, of which 178 full-text articles fulfilled screening criteria and were assessed. Aggregate data on 1185 patients from nine studies were analysed, and individual patient-level data on 895 patients were provided from six of these studies. Eight studies were observational and one was a randomised controlled trial. Most studies used historical controls. In the individual patient-level meta-analysis, after adjusting for age, comorbidities, baseline ratio of the arterial partial oxygen pressure divided by the fraction of inspired oxygen (PaO2/FiO2), C-reactive protein (CRP) concentrations, and lymphopenia, mortality was significantly lower in patients treated with anakinra (38 [11%] of 342) than in those receiving standard of care with or without placebo (137 [25%] of 553; adjusted odds ratio [OR] 0·32 [95% CI 0·20-0·51]). The mortality benefit was similar across subgroups regardless of comorbidities (ie, diabetes), ferritin concentrations, or the baseline PaO2/FiO2. In a subgroup analysis, anakinra was more effective in lowering mortality in patients with CRP concentrations higher than 100 mg/L (OR 0·28 [95% CI 0·17-0·47]). Anakinra showed a significant survival benefit when given without dexamethasone (OR 0·23 [95% CI 0·12-0·43]), but not with dexamethasone co-administration (0·72 [95% CI 0·37-1·41]). Anakinra was not associated with a significantly increased risk of secondary infections when compared with standard of care (OR 1·35 [95% CI 0·59-3·10]). Anakinra could be a safe, anti-inflammatory treatment option to reduce the mortality risk in patients admitted to hospital with moderate to severe COVID-19 pneumonia, especially in the presence of signs of hyperinflammation such as CRP concentrations higher than 100 mg/L. Sobi.


In vitro investigation of anticancer and ACE-inhibiting activity, α-amylase and α-glucosidase inhibition, and antioxidant activity of camel milk fermented with camel milk probiotic: A comparative study with fermented bovine milk.
This study aimed to investigate in vitro the health-promoting benefits (anticancer activity, α-amylase and α-glucosidase inhibition, angiotensin-converting-enzyme (ACE)-inhibition, antioxidant and proteolytic activity) of camel milk fermented with indigenous probiotic strains of Lactobacillus spp., compared with fermented bovine milk. The three camel milk probiotic strains Lb. reuteri-KX881777, Lb. plantarum-KX881772, Lb. plantarum-KX881779 and a control strain Lb. plantarum DSM2468 were employed to ferment camel and bovine milks separately. The proteolytic and antioxidant activity of water soluble extracts (WSEs) from all fermented camel milks were higher than those of fermented bovine milk. α-Amylase inhibition of WSEs were >34% in both milk types fermented with all strains during storage periods, except the WSE of camel milk fermented by Lp.K772. The highest ACE-inhibition of the WSE from camel milk fermented by Lr.K777 was >80%. The proliferations of Caco-2, MCF-7 and HELA cells were more inhibited when treated with the WSE of fermented camel milk.


Barriers to insecticide-treated mosquito net possession 2 years after a mass free distribution campaign in Luangwa District, Zambia.
Roll Back Malaria set the goal of 100% of households in malaria endemic countries in Africa owning an insecticide-treated mosquito net (ITN) by 2010. Zambia has used mass free distribution campaigns and distribution through antenatal care (ANC) clinics to achieve high coverage. We conducted a probability survey of 801 households in 2008 to assess factors associated with households that lacked an ITN after mass distribution. Community perceptions of barriers to ITN access were also obtained from in-depth interviews with household heads that reported not owning an ITN. Nearly 74% of households in Luangwa district reported owning ≥1 ITN. Logistic regression showed households without a child <5 years old during the ITN distribution campaigns were twice as likely to not have an ITN as those with a child <5 during distribution (Adjusted odds ratio (AOR) = 2.43; 95% confidence interval (CI): 1.67-3.55). Households without a woman who attended an ANC in the past 2 years were more likely to be without ITNs compared to households with a woman who attended an ANC in the past 2 years (AOR = 1.52; 95% CI: 1.04-2.21). In-depth interviews with heads of households without an ITN revealed that old age was a perceived barrier to receiving an ITN during distribution, and that ITNs wore out before they could be replaced. Delivery of a large number of ITNs does not translate directly into 100% household coverage. Due to their design, current ITN distribution strategies may miss households occupied by the elderly and those without children or ANC access. ITN distribution strategies targeting the elderly, those with limited access to distribution points, and others most likely to be missed are necessary if 100% ITN coverage of households is to be achieved.


Nonlinear non-negative component analysis algorithms.
In this paper, general solutions for nonlinear non-negative component analysis for data representation and recognition are proposed. Motivated by a combination of the non-negative matrix factorization (NMF) algorithm and kernel theory, which has lead to a recently proposed NMF algorithm in a polynomial feature space, we propose a general framework where one can build a nonlinear non-negative component analysis method using kernels, the so-called projected gradient kernel non-negative matrix factorization (PGKNMF). In the proposed approach, arbitrary positive definite kernels can be adopted while at the same time it is ensured that the limit point of the procedure is a stationary point of the optimization problem. Moreover, we propose fixed point algorithms for the special case of Gaussian radial basis function (RBF) kernels. We demonstrate the power of the proposed methods in face and facial expression recognition applications.


The Drosophila sanpodo gene controls sibling cell fate and encodes a tropomodulin homolog, an actin/tropomyosin-associated protein.
Notch signaling is required in many invertebrate and vertebrate cells to promote proper cell fate determination. Mutations in sanpodo cause many different neuronal peripheral nervous system precursor cells to generate two identical daughter neurons, instead of a neuron and sibling cell. This phenotype is similar to that observed when Notch function is lost late in embryonic development and opposite to the numb loss-of-function phenotype. Genetic interaction studies show that sanpodo is epistatic to numb. sanpodo encodes a homolog of tropomodulin, an actin/tropomyosin-associated protein. Loss of sanpodo leads to an aberrant F-actin distribution and causes differentiation defects of actin-containing sensory structures. Our data suggest that an actin-based process is involved in Notch signaling.


The expression of bipolar spectrum psychopathology in daily life.
Bipolar psychopathology has traditionally been defined by categorical diagnoses. However, these disorders may simply reflect the extremes of a broader spectrum of clinical and subclinical bipolar psychopathology. The present study examined the validity of the Hypomanic Personality Scale (HPS) as a measure of bipolar spectrum psychopathology in 305 young adults using experience sampling methodology. The participants completed the HPS and were signaled randomly eight times daily for seven days to complete brief questionnaires on their current experiences. High HPS scores were associated with elevated energetic-enthusiasm, irritability, dysphoria, flight of ideas, mild grandiose beliefs, and risky behavior, as well as increased variability in affect in daily life. High HPS scores were also associated with greater reactivity in negative affect and behavior in response to viewing themselves as unsuccessful in their activities. It is not clear to what extent the participants had diagnosable bipolar disorders. The findings support a broader spectrum of bipolar psychopathology and the validity of the HPS as a measure of this construct.


Living in Biological Darkness: Objective Sleepiness and the Pupillary Light Responses Are Affected by Different Metameric Lighting Conditions during Daytime.
Nighttime melatonin suppression is the most commonly used method to indirectly quantify acute nonvisual light effects. Since light is the principal zeitgeber in humans, there is a need to assess its strength during daytime as well. This is especially important since humans evolved under natural daylight but now often spend their time indoors under artificial light, resulting in a different quality and quantity of light. We tested whether the pupillary light response (PLR) could be used as a marker for nonvisual light effects during daytime. We also recorded the wake electroencephalogram to objectively determine changes in daytime sleepiness between different illuminance levels and/or spectral compositions of light. In total, 72 participants visited the laboratory 4 times for 3-h light exposures. All participants underwent a dim-light condition and either 3 metameric daytime light exposures with different spectral compositions of polychromatic white light (100 photopic lux, peak wavelengths at 435 nm or 480 nm, enriched with longer wavelengths of light) or 3 different illuminances (200, 600, and 1200 photopic lux) with 1 metameric lighting condition (peak wavelength at 435 nm or 480 nm; 24 participants each). The results show that the PLR was sensitive to both spectral differences between metameric lighting conditions and different illuminances in a dose-responsive manner, depending on melanopic irradiance. Objective sleepiness was significantly reduced, depending on melanopic irradiance, at low illuminance (100 lux) and showed fewer differences at higher illuminance. Since many people are exposed to such low illuminance for most of their day-living in biological darkness-our results imply that optimizing the light spectrum could be important to improve daytime alertness. Our results suggest the PLR as a noninvasive physiological marker for ambient light exposure effects during daytime. These findings may be applied to assess light-dependent zeitgeber strength and evaluate lighting improvements at workplaces, schools, hospitals, and homes.


Pancreatic cancer surgery: past, present, and future.
The history of pancreatic cancer surgery, though fraught with failure and setbacks, is punctuated by periods of incremental progress dependent upon the state of the art and the mettle of the surgeons daring enough to attempt it. Surgical anesthesia and the aseptic techniques developed during the latter half of the 19(th) century were instrumental in establishing a viable setting for pancreatic surgery to develop. Together, they allowed for bolder interventions and improved survival through the post-operative period. Surgical management began with palliative procedures to address biliary obstruction in advanced disease. By the turn of the century, surgical pioneers such as Alessandro Codivilla and Walther Kausch were demonstrating the technical feasibility of pancreatic head resections and applying principles learned from palliation to perform complicated anatomical reconstructions. Allen O. Whipple, the namesake of the pancreaticoduodenectomy (PD), was the first to take a systematic approach to refining the procedure. Perhaps his greatest contribution was sparking a renewed interest in the surgical management of periampullary cancers and engendering a community of surgeons who advanced the field through their collective efforts. Though the work of Whipple and his contemporaries legitimized PD as an accepted surgical option, it was the establishment of high-volume centers of excellence and a multidisciplinary approach in the later decades of the 20(th) century that made it a viable surgical option. Today, pancreatic surgeons are experimenting with minimally invasive surgical techniques, expanding indications for resection, and investigating new methods for screening and early detection. In the future, the effective management of pancreatic cancer will depend upon our ability to reliably detect the earliest cancers and precursor lesions to allow for truly curative resections.


Comparative Analysis of Fecal Microbiota in Grasscutter (Thryonomys swinderianus) and Other Herbivorous Livestock in Ghana.
The grasscutter (also known as the greater cane rat; Thryonomys swinderianus) is a large rodent native to West Africa that is currently under domestication process for meat production. However, little is known about the physiology of this species. In the present study, aiming to provide information about gut microbiota of the grasscutter and better understand its physiology, we investigated the intestinal microbiota of grasscutters and compared it with that of other livestock (cattle, goat, rabbit, and sheep) using 16S rRNA metagenomics analysis. Similar to the other herbivorous animals, bacteria classified as Bacteroidales, Clostridiales, Ruminococcaceae, and Lachnospiraceae were abundant in the microbiome of grasscutters. However, Prevotella and Treponema bacteria, which have fiber fermentation ability, were especially abundant in grasscutters, where the relative abundance of these genera was higher than that in the other animals. The presence of these genera might confer grasscutters the ability to easily breakdown dietary fibers. Diets for grasscutters should be made from ingredients not consumed by humans to avoid competition for resources and the ability to digest fibers may allow the use of fiber-rich feed materials not used by humans. Our findings serve as reference and support future studies on changes in the gut microbiota of the grasscutter as domestication progresses in order to establish appropriate feeding methods and captivity conditions.


Selective alterations of white matter associated with visuospatial and sensorimotor dysfunction in turner syndrome.
Turner syndrome (TS) is a neurogenetic disorder characterized by impaired spatial, numerical, and motor functioning but relatively spared verbal ability. Results from previous neuroimaging studies suggest that gray matter alterations in parietal and frontal regions may contribute to atypical visuospatial and executive functioning in TS. Recent findings in TS also indicate variations in the shape of parietal gyri and white matter microstructural anomalies of the temporal lobe. Diffusion tensor imaging and structural imaging methods were used to determine whether 10 females with TS and 10 age- and gender-matched control subjects exhibited differences in fractional anisotropy, white matter density, and local brain shape. Relative to controls, females with TS had lower fractional anisotropy (FA) values in the deep white matter of the left parietal-occipital region extending anteriorly along the superior longitudinal fasciculus into the deep white matter of the frontal lobe. In addition, decreased FA values were located bilaterally in the internal capsule extending into the globus pallidus and in the right prefrontal region. Voxel-based morphometry (VBM) analysis showed corresponding white matter density differences in the internal capsules and left centrum semiovale. Tensor-based morphometry analysis indicated that the FA and VBM results were not attributable to differences in the local shape of brain structures. Compared with controls, females with TS had increases in FA values and white matter density in language-related areas of the inferior parietal and temporal lobes. These complementary analyses provide evidence for alterations in white matter pathways that subserve affected and preserved cognitive functions in TS.


Deep Bite Malocclusion: Exploration Of The Skeletal And Dental Factors.
Correction of deep bite is crucial for maintenance of dental hard and soft tissue structures and for prevention of temporomandibular joint disorders. Exploration of underlying skeletal and dental factors is essential for efficient and individualized treatment planning. To date etiological factors of dental and skeletal deep bite have not been explored in Pakistani orthodontic patients. The objectives of this study were to explore frequencies of dental and skeletal etiological factors in deep bite patients and to determine correlations amongst dental and skeletal etiological factors of deep bite. The study included a total of 113 subjects (males=35; females=78) with no craniofacial syndromes or prior orthodontic treatment. Pre-treatment orthodontic records were used to evaluate various dental and skeletal parameters. Descriptive statistics of each parameter were calculated. The various study parameters were correlated using Pearson's Correlation. Deep curve of Spee was most frequently seen factor of dental deep bite (72.6%), followed by increased coronal length of upper incisors (28.3%), retroclined upper incisors (17.7%), retroclined lower incisors (8%) and increased coronal length of lower incisors (5.3%). Decreased gonial angle was most commonly found factor of skeletal deep bite (43.4%), followed by decreased mandibular plane angle (27.4%) and maxillary plane's clockwise rotation (26.5%). Frankfort mandibular plane angle and gonial angle showed a strong positive correlation (r=0.66, p=0.000). Reduced gonial angle is most frequently seen skeletal factor, signifying the importance of angulation and growth of ramus in development of deep bite. Deep curve of Spee is most frequently seen dental etiological component in deep bite subjects, hence signifying the importance of intruding the lower anterior teeth.


A common new species of Inocybe in the Pacific Northwest with a diagnostic PDAB reaction.
A species of Inocybe common in Washington, Oregon and British Columbia is documented and described as new. The species, I. chondroderma, is characterized by these features: pileus with a fulvous disk and ochraceous to chamois margin, presence of a cortina, densely mycelioid stipe base, smooth spores and fall phenology. The most reliable and distinctive feature of the species is a blue-green or turquoise reaction in response to application of a solution of p-dimethylaminobenzaldehyde (PDAB), indicating the presence of what is most likely an indole alkaloid. PDAB use provides a quick and diagnostic character easily implemented in a laboratory setting. ITS sequences from recent collections of I. chondroderma and from materials collected in the 1940s in Washington and Oregon fully match numerous mislabeled sequences from specimens in British Columbia and Oregon. The species is most closely related to an unclarified taxon from Colorado and Japan (I. cf. chondroderma) and a rare European species, I. subnudipes. Nine different species names in Inocybe and one in Hebeloma attributed to I. chondroderma based on GenBank BLASTN searches of the ITS locus match with 99-100% similarity, reinforcing concerns about taxonomic inaccuracies in public DNA sequence databases. A complete morphological description, illustrations and phylogenetic assessment are provided.


Should we end military recruiting in high schools as a matter of child protection and public health?
Recruiters for the various US armed forces have free access to our nation's high schools, as mandated by the No Child Left Behind Act. Military recruiter behaviors are disturbingly similar to predatory grooming. Adults in the active military service are reported to experience increased mental health risks, including stress, substance abuse, and suicide, and the youngest soldiers consistently show the worst health effects, suggesting military service is associated with disproportionately poor health for this population. We describe the actions of a high school parent teacher student association in Seattle, Washington, which sought to limit the aggressive recruitment of children younger than 18 years into the military.


Context and time in causal learning: contingency and mood dependent effects.
Defining cues for instrumental causality are the temporal, spatial and contingency relationships between actions and their effects. In this study, we carried out a series of causal learning experiments that systematically manipulated time and context in positive and negative contingency conditions. In addition, we tested participants categorized as non-dysphoric and mildly dysphoric because depressed mood has been shown to affect the processing of all these causal cues. Findings showed that causal judgements made by non-dysphoric participants were contextualized at baseline and were affected by the temporal spacing of actions and effects only with generative, but not preventative, contingency relationships. Participants categorized as dysphoric made less contextualized causal ratings at baseline but were more sensitive than others to temporal manipulations across the contingencies. These effects were consistent with depression affecting causal learning through the effects of slowed time experience on accrued exposure to the context in which causal events took place. Taken together, these findings are consistent with associative approaches to causal judgement.


Time to change weight gain recommendations for pregnant women with obesity.
Obesity during pregnancy is a major health problem in the United States. In this issue of the JCI, Most et al. fill an important gap in our understanding of energy homeostasis in pregnancy. The researchers measured energy intake, energy expenditure, and body composition in obese pregnant women. They demonstrated that energy intake need not increase in order for obese women to gain the recommended amounts of weight during pregnancy. Additionally, all of the gestational weight gain scenarios (inadequate, recommended, or excess) resulted in similar maternal and fetal perinatal outcomes. This evidence should guide new recommendations on this important topic.


TLR agonists abrogate costimulation blockade-induced prolongation of skin allografts.
Costimulation blockade protocols are effective in prolonging allograft survival in animal models and are entering clinical trials, but how environmental perturbants affect graft survival remains largely unstudied. We used a costimulation blockade protocol consisting of a donor-specific transfusion and anti-CD154 mAb to address this question. We observed that lymphocytic choriomeningitis virus infection at the time of donor-specific transfusion and anti-CD154 mAb shortens allograft survival. Lymphocytic choriomeningitis virus 1) activates innate immunity, 2) induces allo-cross-reactive T cells, and 3) generates virus-specific responses, all of which may adversely affect allograft survival. To investigate the role of innate immunity, mice given costimulation blockade and skin allografts were coinjected with TLR2 (Pam3Cys), TLR3 (polyinosinic:polycytidylic acid), TLR4 (LPS), or TLR9 (CpG) agonists. Costimulation blockade prolonged skin allograft survival that was shortened after coinjection by TLR agonists. To investigate underlying mechanisms, we used "synchimeric" mice which circulate trace populations of anti-H2b transgenic alloreactive CD8+ T cells. In synchimeric mice treated with costimulation blockade, coadministration of all four TLR agonists prevented deletion of alloreactive CD8+ T cells and shortened skin allograft survival. These alloreactive CD8+ T cells 1) expressed the proliferation marker Ki-67, 2) up-regulated CD44, and 3) failed to undergo apoptosis. B6.TNFR2-/- and B6.IL-12R-/- mice treated with costimulation blockade plus LPS also exhibited short skin allograft survival whereas similarly treated B6.CD8alpha-/- and TLR4-/- mice exhibited prolonged allograft survival. We conclude that TLR signaling abrogates the effects of costimulation blockade by preventing alloreactive CD8+ T cell apoptosis through a mechanism not dependent on TNFR2 or IL-12R signaling.


Graft rejection-like reactions in the early postoperative period after deep anterior lamellar keratoplasty for keratoconus: a retrospective study.
Deep anterior lamellar keratoplasty (DALK) is indicated to correct high astigmatism in patients with keratoconus (KC) and no evidence of Descemet's membrane rupture. However, some patients with KC experience graft rejection-like inflammatory reactions within 2 months (usually in the first week) after DALK. The aim of this study was to identify the characteristics and influencing factors of these reactions by reviewing the records of patients who underwent DALK or penetrating keratoplasty (PKP) for KC or other corneal problems. We retrospectively reviewed the medical records of patients who underwent DALK for KC (DALK/KC), PKP for KC (PKP/KC), or DALK for other corneal problems (DALK/non-KC) at Ban Buntane Hotokukai Hospital between January 2006 and December 2015 and who were followed for more than 1 year. We collected data on the characteristics and incidence of severe inflammatory graft reactions in the early postoperative phase (ie, within 2 months after keratoplasty) and visual outcomes after these inflammatory reactions. Postoperative inflammatory reactions characterized by persistent epithelial defects, loose suture with infiltration, and vessel invasion occurred in seven of eleven DALK/KC patients, three of 50 DALK/non-KC patients, and none of five PKP/KC patients. These reactions were nonresponsive to topical steroids, and suture removal was required. Although a clear corneal graft in the pupillary area was obtained and best-corrected visual acuity was good after the resolution of inflammation, a risk of corneal astigmatism remained. The incidence of these reactions in the early postoperative period is high after DALK for KC. These reactions do not respond well to topical steroids, and suture removal may be required, which may cause high astigmatism after the inflammation subsides. Lamellar keratoplasty should be considered carefully for patients with KC.


Performance of the Breast Cancer Risk Assessment Tool Among Women Age 75 Years and Older.
The Breast Cancer Risk Assessment Tool (BCRAT, "Gail model") is commonly used for breast cancer prediction; however, it has not been validated for women age 75 years and older. We used Nurses' Health Study (NHS) data beginning in 2004 and Women's Health Initiative (WHI) data beginning in 2005 to compare BCRAT's performance among women age 75 years and older with that in women age 55 to 74 years in predicting five-year breast cancer incidence. BCRAT risk factors include: age, race/ethnicity, age at menarche, age at first birth, family history, history of benign breast biopsy, and atypia. We examined BCRAT's calibration by age by comparing expected/observed (E/O) ratios of breast cancer incidence. We examined discrimination by computing c-statistics for the model by age. All statistical tests were two-sided. Seventy-three thousand seventy-two NHS and 97 081 WHI women participated. NHS participants were more likely to be non-Hispanic white (96.2% vs 84.7% in WHI, P < .001) and were less likely to develop breast cancer (1.8% vs 2.0%, P = .02). E/O ratios by age in NHS were 1.16 (95% confidence interval [CI] = 1.09 to 1.23, age 57-74 years) and 1.31 (95% CI = 1.18 to 1.45, age ≥ 75 years, P = .02), and in WHI 1.03 (95% CI = 0.97 to 1.09, age 55-74 years) and 1.10 (95% CI = 1.00 to 1.21, age ≥ 75 years, P = .21). E/O ratio 95% confidence intervals crossed one among women age 75 years and older when samples were limited to women who underwent mammography and were without significant illness. C-statistics ranged between 0.56 and 0.58 in both cohorts regardless of age. BCRAT accurately predicted breast cancer for women age 75 years and older who underwent mammography and were without significant illness but had modest discrimination. Models that consider individual competing risks of non-breast cancer death may improve breast cancer risk prediction for older women.


Entrapped in cage (EiC) scaffolds of 3D-printed polycaprolactone and porous silk fibroin for meniscus tissue engineering.
The meniscus has critical functions in the knee joint kinematics and homeostasis. Injuries of the meniscus are frequent, and the lack of a functional meniscus between the femur and tibial plateau can cause articular cartilage degeneration leading to osteoarthritis development and progression. Regeneration of meniscus tissue has outstanding challenges to be addressed. In the current study, novel Entrapped in cage (EiC) scaffolds of 3D-printed polycaprolactone (PCL) and porous silk fibroin were proposed for meniscus tissue engineering. As confirmed by micro-structural analysis the entrapment of silk fibroin was successful, and all scaffolds had excellent interconnectivity (≥99%). The EiC scaffolds had more favorable micro-structure compared with the PCL cage scaffolds by improving the pore size while keeping the interconnectivity almost the same. When compared with the PCL cage, the entrapment of porous silk fibroin into the PCL cage decreased the high compressive modulus in a favorable matter in the wet state thanks to the silk fibroin's high swelling properties. The in vitro studies with human stem cells or meniscocytes seeded constructs, demonstrated that the EiC scaffolds had superior cell adhesion, metabolic activity, and proliferation compared to the PCL cage scaffolds. Upon subcutaneous implantation of scaffolds in nude mice, all groups were free of adverse incidents, and mildly invaded by inflammatory cells with neovascularization, while the EiC scaffolds showed better tissue infiltration. The results of this work indicated that the EiC scaffolds of PCL and silk fibroin are favorable for meniscus tissue engineering, and the findings are encouraging for further studies using a larger animal model.


Hong-Ou-Mandel Interference between Two Deterministic Collective Excitations in an Atomic Ensemble.
We demonstrate deterministic generation of two distinct collective excitations in one atomic ensemble, and we realize the Hong-Ou-Mandel interference between them. Using Rydberg blockade we create single collective excitations in two different Zeeman levels, and we use stimulated Raman transitions to perform a beam-splitter operation between the excited atomic modes. By converting the atomic excitations into photons, the two-excitation interference is measured by photon coincidence detection with a visibility of 0.89(6). The Hong-Ou-Mandel interference witnesses an entangled NOON state of the collective atomic excitations, and we demonstrate its two times enhanced sensitivity to a magnetic field compared with a single excitation. Our work implements a minimal instance of boson sampling and paves the way for further multimode and multiexcitation studies with collective excitations of atomic ensembles.


Greater three-dimensional ventricular lead tip separation is associated with improved outcome after cardiac resynchronization therapy.
Effective cardiac resynchronization therapy (CRT) is more likely with widely separated left ventricular (LV) and right ventricular (RV) pacing leads tips. We hypothesized that lead separation is an important factor in determining the clinical response to CRT. A retrospective study of 86 consecutive patients age 71 ± 10 years, male (74%), coronary disease (71%), atrial fibrillation (23%), LV ejection fraction (22 ± 9%), QRS duration (160 ± 27 ms), New York Heart Association (NYHA) class III (81%), NYHA class IV (19%) undergoing CRT from January 2006 to September 2008. The median follow-up was 12 months and clinical response to CRT was defined as reduction of NYHA class by one or more. The three-dimensional separation between RV and LV pacing lead tips was calculated using measurements obtained from orthogonal posteroanterior and lateral chest radiographs performed the day after implantation. Fifty-nine patients (69%) responded to CRT. There was a statistically significant association between increased three-dimensional lead separation and clinical response to CRT (P= 0.005). Stronger association was obtained when lead separation was corrected for cardiac size (P= 0.001). A significantly higher response rate of 88% was achieved in patients with QRS duration of 160 ms or more, and lead separation of 100 mm or more compared with 60% when lead separation was less than 100 mm and QRS duration remained the same (P = 0.027). Greater three-dimensional separation of LV-to-RV leads is associated with improved response to CRT. A prospective multicenter trial is needed to assess lead separation as a predictor for response.


Variable fate of virus-specific CD4(+) T cells during primary HIV-1 infection.
Impairment of CD4(+) T lymphocyte responses to human immunodeficiency virus (HIV)-derived antigens is the classic immunological defect observed during the chronic phase of HIV-1 infection. Early intervention with potent antiretroviral therapy (ART) can preserve HIV-specific CD4(+) T lymphocyte reactivity, providing indirect evidence that such responses are mounted during primary infection and subsequently lost in the majority of infected individuals. Here, we demonstrate early and dramatic expansions of functional HIV-specific CD4(+) T lymphocyte frequencies directly ex vivo. These responses are initially of broad specificity, and can disappear rapidly during the natural course of primary infection. This process of loss is variable, such that the rapidity and extent of functional compromise differs between individuals. Institution of ART during these early phases of HIV-1 infection preserves patterns of functional reactivity within the HIV-specific CD4(+) T lymphocyte population. However, there was no evidence for the restoration of deleted responses. These findings indicate that, in some individuals at least, ART must be administered within a narrow window of opportunity during primary HIV-1 infection to effect substantial immune preservation.


A preliminary survey of the median artery in human cadavers of South Indian origin.
The median artery is a transitory vessel that represents the arterial axis of the forearm during early embryonic life. It normally regresses in the second embryonic month. Its persistence in the human adult has been recorded in 2 different patterns: as a large, long vessel (palmar type) which reaches the hand; or as a small and short vessel (antebrachial type) which ends before reaching the wrist joint. The palmar type is of major clinical significance. This study was undertaken to investigate the incidence and course of the palmar type of the median artery in South Indian cadavers. 25 upper limbs of South Indian cadavers were taken to study the median artery. The occurrence of median artery was 8%; of which 4% was on the right side and the other 4% was on the left side. On both sides, the artery originated from the ulnar artery. On the right side, the artery was involved in the formation of superficial palmar arch, whereas the artery on the left side did not join the arch; it terminated as 1st and 2nd common palmar digital arteries. Persistent median artery is closely related to the anterior interosseous nerve, it is possible that the artery may compress the anterior interosseous nerve and cause the anterior interosseous nerve syndrome (Fig. 2, Ref. 17).


Improvement in Vision Parameters for Participants Treated With Alternative Therapies in a 3-day Program.
Eye conditions that are considered progressive and degenerative and for which the causation is generally poorly understood or not understood within conventional medicine can respond to natural therapeutic interventions that result in arrest and/or improvement of morbidity, with enhanced functional results. Because many of the treated conditions are age related, a delay of disease progression for 5 or even 10 y can mean an additional decade of independence for seniors. The 11 included ocular conditions are ordinarily considered incurable by any method except surgery and, even with surgery, the outcomes can be variable and/or transient. The research intended to demonstrate the effectiveness of alternative modalities-intravenous (IV) nutrition, oxidative therapy, microcurrent stimulation, and syntonic light therapy-in improving vision in chronic eye conditions, even when administered for a short period. The study was a retrospective, open-label, single-group design. All participants in the 3-d conference during the period covered were selected. The setting was ophthalmologist Edward Kondrot's Healing the Eye and Wellness Center near Tampa, FL, USA. The participants in this study were all patients attending 1 of 11 CAM treatment events at the author's center within 2 y. Each session lasted 3 d and the number of participants in each session ranged from 5-15 (mean = 13). The cohort numbered 152 patients who were diagnosed with ≥1 of 11 types of eye disease. Seventy-eight percent of the patients had either age-related macular degeneration (ARMD) or glaucoma, which, taken together, are the leading cause of blindness in persons >65 y. Each of 4 alternative modalities was provided at least once to each participant: (1) IV nutrition, (2) oxidative therapy, (3) microcurrent stimulation, and (4) syntonic light therapy. On the first day, a detailed treatment plan for each participant was developed. Each day consisted of 2 therapeutic eye programs, a stress reduction program, and a detoxification program. Also included were daily lectures and instructions on the methods and use of the equipment. To measure outcomes, changes from baseline were documented through comparison with postprogram results. Pre- and postprogram testing included the following measures: (1) Early Treatment Diabetic Retinopathy Study (ETDRS) eye chart; (2) Lighthouse Letter Contrast Sensitivity test; (3) campimetry; (4) pursuits, saccade, and fixation tests; (5) pupillary examination; (6) external examination; (7) examination of the anterior segment; (8) intraocular-pressure test; and (9) dilated examination. Additional tests, if necessary, included (1) ocular coherence tomography, (2) infrared thermography, (3) 6-hour urine collection for heavy-metal toxicity, and (4) nocturnal oximetry. All participants remained in the study for the duration of the program. Following the administration of the protocol, significant improvement in acuity, contrast, and visual field resulted in the majority of participants. None of the interventions was toxic or painful, and all likely contributed to an improved, overall health status for participants. These treatment protocols should be considered part of a treatment program for all ocular disease processes. Eye health needs to be repositioned within an assessment of general health with the understanding that, with the exception of congenital disorders or accidents, vision decline represents a general diminishment in overall health and results directly from toxicity from both external sources such as air and water, and the internal accumulation of toxic metals; poor nutrition; and other life exposures and habits. Long-term follow-up studies are now in process.


Muscle hypertrophy response to resistance training in older women.
We conducted a 12-wk resistance training program in elderly women [mean age 69 +/- 1.0 (SE) yr] to determine whether increases in muscle strength are associated with changes in cross-sectional fiber area of the vastus lateralis muscle. Twenty-seven healthy women were randomly assigned to either a control or exercise group. The program was satisfactorily completed and adequate biopsy material obtained from 6 controls and 13 exercisers. After initial testing of baseline maximal strength, exercisers began a training regimen consisting of seven exercises that stressed primary muscle groups of the lower extremities. No active intervention was prescribed for the controls. Increases in muscle strength of the exercising subjects were significant compared with baseline values (28-115%) in all muscle groups. No significant strength changes were observed in the controls. Cross-sectional area of type II muscle fibers significantly increased in the exercisers (20.1 +/- 6.8%, P = 0.02) compared with baseline. In contrast, no significant change in type II fiber area was observed in the controls. No significant changes in type I fiber area were found in either group. We conclude that a program of resistance exercise can be safely carried out by elderly women, such a program significantly increases muscle strength, and such gains are due, at least in part, to muscle hypertrophy.


Irradiated sporozoite vaccine induces HLA-B8-restricted cytotoxic T lymphocyte responses against two overlapping epitopes of the Plasmodium falciparum sporozoite surface protein 2.
Vaccines designed to protect against malaria by inducing CD8+ cytotoxic T lymphocytes (CTL) in individuals of diverse HLA backgrounds must contain multiple conserved epitopes from various preerythrocytic-stage antigens. Plasmodium falciparum sporozoite surface protein 2 (PfSSP2) is considered an important antigen for inclusion in such vaccines, because CD8+ CTL against the P. yoelii SSP2 protect mice against malaria by eliminating infected hepatocytes. To develop PfSSP2 as a component of malaria vaccines, we investigated the presence of anti-PfSSP2 CTL in two HLA-B8+ volunteers immunized with irradiated P. falciparum sporozoites and characterized their CTL responses using PfSSP2-derived 15-amino acid peptides bearing the HLA-B8-binding motif. Peripheral blood mononuclear cells from both volunteers stimulated with recombinant vaccinia expressing PfSSP2 displayed antigen-specific, genetically restricted, CD8+ T cell-dependent CTL activity against autologous target cells expressing PfSSP2. Of the five HLA-B8 motif-bearing 15-mers identified in the PfSSP2 sequence, two peptides sharing a 10-amino acid overlap sensitized HLA-B8-matched target cells from both volunteers for lysis by peptide-stimulated effectors. The CTL activity was HLA-B8 restricted and dependent on CD8+ T cells. Analysis of the three shorter peptides representing HLA-B8 motif-bearing sequences within the two positive peptides for their ability to bind to HLA-B8 in vitro, and to sensitize target cells for lysis by effectors stimulated with the 15-mers, identified two overlapping HLA-B8-restricted CTL epitopes. Available data indicate that the sequence of one CTL epitope is conserved and the other is variant among P. falciparum isolates. Circulating activated CTL against the conserved epitope could be directly identified in one of the two volunteers. The identification of two HLA-B8-restricted CTL epitopes on PfSSP2 provides data critical to developing an epitope-based anti-liver stage malaria vaccine.


Parallel visual search and rapid animal detection in natural scenes.
Human observers are capable of detecting animals within novel natural scenes with remarkable speed and accuracy. Recent studies found human response times to be as fast as 120 ms in a dual-presentation (2-AFC) setup (H. Kirchner & S. J. Thorpe, 2005). In most previous experiments, pairs of randomly chosen images were presented, frequently from very different contexts (e.g., a zebra in Africa vs. the New York Skyline). Here, we tested the effect of background size and contiguity on human performance by using a new, contiguous background image set. Individual images contained a single animal surrounded by a large, animal-free image area. The image could be positioned and cropped in such a manner that the animal could occur in one of eight evenly spaced positions on an imaginary circle (radius 10-deg visual angle). In the first (8-Choice) experiment, all eight positions were used, whereas in the second (2-Choice) and third (2-Image) experiments, the animals were only presented on the two positions to the left and right of the screen center. In the third experiment, additional rectangular frames were used to mimic the conditions of earlier studies. Average latencies on successful trials differed only slightly between conditions, indicating that the number of possible animal locations within the display does not affect decision latency. Detailed analysis of saccade targets revealed a preference toward both the head and the center of gravity of the target animal, affecting hit ratio, latency, and the number of saccades required to reach the target. These results illustrate that rapid animal detection operates scene-wide and is fast and efficient even when the animals are embedded in their natural backgrounds.


Spiropyrimidinetrione DNA Gyrase Inhibitors with Potent and Selective Antituberculosis Activity.
New antibiotics with either a novel mode of action or novel mode of inhibition are urgently needed to overcome the threat of drug-resistant tuberculosis (TB). The present study profiles new spiropyrimidinetriones (SPTs), DNA gyrase inhibitors having activity against drug-resistant Mycobacterium tuberculosis (Mtb), the causative agent of TB. While the clinical candidate zoliflodacin has progressed to phase 3 trials for the treatment of gonorrhea, compounds herein demonstrated higher inhibitory potency against Mtb DNA gyrase (e.g., compound 42 with IC50 = 2.0) and lower Mtb minimum inhibitor concentrations (0.49 μM for 42). Notably, 42 and analogues showed selective Mtb activity relative to representative Gram-positive and Gram-negative bacteria. DNA gyrase inhibition was shown to involve stabilization of double-cleaved DNA, while on-target activity was supported by hypersensitivity against a gyrA hypomorph. Finally, a docking model for SPTs with Mtb DNA gyrase was developed, and a structural hypothesis was built for structure-activity relationship expansion.


Pediatric antimicrobial susceptibility trends across the United States.
Antimicrobial susceptibility patterns across US pediatric healthcare institutions are unknown. A national pooled pediatric antibiogram (1) identifies nationwide trends in antimicrobial resistance, (2) allows across-hospital benchmarking, and (3) provides guidance for empirical antimicrobial regimens for institutions unable to generate pediatric antibiograms. In January 2012, a request for submission of pediatric antibiograms between 2005 and 2011 was sent to 233 US hospitals. A summary antibiogram was compiled from participating institutions to generate proportions of antimicrobial susceptibility. Temporal and regional comparisons were evaluated using χ(2) tests and logistic regression, respectively. Of 200 institutions (85%) responding to our survey, 78 (39%) reported generating pediatric antibiograms, and 55 (71%) submitted antibiograms. Carbapenems had the highest activity against the majority of gram-negative organisms tested, but no antibiotic had more than 90% activity against Pseudomonas aeruginosa. Approximately 50% of all Staphylococcus aureus isolates were methicillin resistant. Western hospitals had significantly lower proportions of S. aureus that were methicillin resistant compared with all other regions tested. Overall, 21% of S. aureus isolates had resistance to clindamycin. Among Enterococcus faecium isolates, the prevalence of susceptibility to ampicillin (25%) and vancomycin (45%) was low but improved over time (P< .01), and 8% of E. faecium isolates were resistant to linezolid. Southern hospitals reported significantly higher prevalence of E. faecium with susceptibilities to ampicillin, vancomycin, and linezolid compared with the other 3 regions (P< .01). A pooled, pediatric antibiogram can identify nationwide antimicrobial resistance patterns for common pathogens and might serve as a useful tool for benchmarking resistance and informing national prescribing guidelines for children.


Simulating the impact of long-term care policy on family eldercare hours.
To understand the effect of current and future long-term care (LTC) policies on family eldercare hours for older adults (60 years of age and older) in Singapore. The Social Isolation Health and Lifestyles Survey, the Survey on Informal Caregiving, and the Singapore Government's Ministry of Health and Department of Statistics. An LTC Model was created using system dynamics methodology and parameterized using available reports and data as well as informal consultation with LTC experts. In the absence of policy change, among the elderly living at home with limitations in their activities of daily living (ADLs), the proportion of those with greater ADL limitations will increase. In addition, by 2030, average family eldercare hours per week are projected to increase by 41 percent from 29 to 41 hours. All policy levers considered would moderate or significantly reduce family eldercare hours. System dynamics modeling was useful in providing policy makers with an overview of the levers available to them and in demonstrating the interdependence of policies and system components.


A larger brain confers a benefit in a spatial mate search learning task in male guppies.
Brain size varies dramatically among vertebrates, and selection for increased cognitive abilities is thought to be the key force underlying the evolution of a large brain. Indeed, numerous comparative studies suggest positive relationships between cognitively demanding aspects of behavior and brain size controlled for body size. However, experimental evidence for the link between relative brain size and cognitive ability is surprisingly scarce and to date stems from a single study on brain size selected guppies (Poecilia reticulata), where large-brained females were shown to outperform small-brained females in a numerical learning assay. Because the results were inconclusive for males in that study, we here use a more ecologically relevant test of male cognitive ability to investigate whether or not a relatively larger brain increases cognitive ability also in males. We compared mate search ability of these artificially selected large- and small-brained males in a maze and found that large-brained males were faster at learning to find a female in a maze. Large-brained males decreased the time spent navigating the maze faster than small-brained males and were nearly twice as fast through the maze after 2 weeks of training. Our results support that relatively larger brains are better also for males in some contexts, which further substantiates that variation in vertebrate brain size is generated through the balance between energetic costs and cognitive benefits.


Quantitative ultrasound in relation to risk factors for low bone mineral density in South African pre-menopausal women.
SUMMARY: The study describes the association between risk factors and quantitative ultrasound bone measures in black and mixed-race pre-menopausal South African women. Despite some differences between the two study groups, the findings generally lend support to the use of ultrasound for epidemiological studies of bone mass in resource-limited settings. INTRODUCTION: Quantitative ultrasound at the calcaneus is a convenient and inexpensive method of estimating bone strength well suited to community-based research in countries with limited resources. This study determines, in a large sample of pre-menopausal South African women, whether characteristics associated with quantitative ultrasound measures are similar to those shown to be associated with bone mineral density as measured by dual X-ray absorptiometry. METHODS: This cross-sectional study included 3,493 women (1,598 black and 1,895 mixed race), aged 18-44 living in Cape Town. Study nurses administered structured interviews on reproductive history, lifestyle factors, and measured height and weight. Calcaneus quantitative ultrasound measurements were obtained using the Sahara device. Adjusted means of ultrasound measures according to categories of risk factors were obtained using multivariable regression analysis. RESULTS: Associations between quantitative ultrasound measures and age, body mass index, age at menarche, parity, and primary school physical activity were similar to those known for bone mineral density as measured by dual X-ray absorptiometry. There were no clear associations between quantitative ultrasound measures and educational level, alcohol use, cigarette smoking, and current calcium intake. CONCLUSION: The data give qualified support to the use of quantitative ultrasound as an epidemiological tool in large studies of bone strength in pre-menopausal women.


Switching the Post-translational Modification of Translation Elongation Factor EF-P.
Tripeptides with two consecutive prolines are the shortest and most frequent sequences causing ribosome stalling. The bacterial translation elongation factor P (EF-P) relieves this arrest, allowing protein biosynthesis to continue. A seven amino acids long loop between beta-strands β3/β4 is crucial for EF-P function and modified at its tip by lysylation of lysine or rhamnosylation of arginine. Phylogenetic analyses unveiled an invariant proline in the -2 position of the modification site in EF-Ps that utilize lysine modifications such as Escherichia coli. Bacteria with the arginine modification like Pseudomonas putida on the contrary have selected against it. Focusing on the EF-Ps from these two model organisms we demonstrate the importance of the β3/β4 loop composition for functionalization by chemically distinct modifications. Ultimately, we show that only two amino acid changes in E. coli EF-P are needed for switching the modification strategy from lysylation to rhamnosylation.


Association of Baseline Visual Acuity and Retinal Thickness With 1-Year Efficacy of Aflibercept, Bevacizumab, and Ranibizumab for Diabetic Macular Edema.
Comparisons of the relative effect of 3 anti-vascular endothelial growth factor agents to treat diabetic macular edema warrant further assessment. To provide additional outcomes from a randomized trial evaluating 3 anti-vascular endothelial growth factor agents for diabetic macular edema within subgroups based on baseline visual acuity (VA) and central subfield thickness (CST) as evaluated on optical coherence tomography. Post hoc exploratory analyses were conducted of randomized trial data on 660 adults with diabetic macular edema and decreased VA (Snellen equivalent, approximately 20/32 to 20/320). The original study was conducted between August 22, 2012, and August 28, 2013. Analysis was conducted from January 7 to June 2, 2015. Repeated 0.05-mL intravitreous injections of 2.0 mg of aflibercept (224 eyes), 1.25 mg of bevacizumab (218 eyes), or 0.3 mg of ranibizumab (218 eyes) as needed per protocol. One-year VA and CST outcomes within prespecified subgroups based on both baseline VA and CST thresholds, defined as worse (20/50 or worse) or better (20/32 to 20/40) VA and thicker (≥400 µm) or thinner (250 to 399 µm) CST. In the subgroup with worse baseline VA (n = 305), irrespective of baseline CST, aflibercept showed greater improvement than bevacizumab or ranibizumab for several VA outcomes. In the subgroup with better VA and thinner CST at baseline (61-73 eyes across 3 treatment groups), VA outcomes showed little difference between groups; mean change was +7.2, +8.4, and +7.6 letters in the aflibercept, bevacizumab, and ranibizumab groups, respectively. However, in the subgroup with better VA and thicker CST at baseline (31-43 eyes), there was a suggestion of worse VA outcomes in the bevacizumab group; mean change from baseline to 1 year was +9.5, +5.4, and +9.5 letters in the aflibercept, bevacizumab, and ranibizumab groups, respectively, and VA letter score was greater than 84 (approximately 20/20) in 21 of 33 (64%), 7 of 31 (23%), and 21 of 43 (49%) eyes, respectively. The adjusted differences and 95% CIs were 39% (17% to 60%) for aflibercept vs bevacizumab, 25% (5% to 46%) for ranibizumab vs bevacizumab, and 13% (-8% to 35%) for aflibercept vs ranibizumab. These post hoc secondary findings suggest that for eyes with better initial VA and thicker CST, some VA outcomes may be worse in the bevacizumab group than in the aflibercept and ranibizumab groups. Given the exploratory nature of these analyses and the small sample size within subgroups, caution is suggested when using the data to guide treatment considerations for patients. clinicaltrials.gov Identifier: NCT01627249.


REGEN: Ancestral Genome Reconstruction for Bacteria.
Ancestral genome reconstruction can be understood as a phylogenetic study with more details than a traditional phylogenetic tree reconstruction. We present a new computational system called REGEN for ancestral bacterial genome reconstruction at both the gene and replicon levels. REGEN reconstructs gene content, contiguous gene runs, and replicon structure for each ancestral genome. Along each branch of the phylogenetic tree, REGEN infers evolutionary events, including gene creation and deletion and replicon fission and fusion. The reconstruction can be performed by either a maximum parsimony or a maximum likelihood method. Gene content reconstruction is based on the concept of neighboring gene pairs. REGEN was designed to be used with any set of genomes that are sufficiently related, which will usually be the case for bacteria within the same taxonomic order. We evaluated REGEN using simulated genomes and genomes in the Rhizobiales order.


Descriptive epidemiology of missed cases of phenylketonuria and congenital hypothyroidism.
We conducted a structured telephone survey of state public health laboratory directors of neonatal screening programs to determine the extent of the problem of missed cases of phenylketonuria (PKU) and congenital hypothyroidism. A total of 76 missed cases were reported--43 PKU and 33 congenital hypothyroidism. We looked at the following characteristics of the missed cases: the stage at which the miss occurred, which included specimen collection, laboratory procedures, or follow-up; the size of the program; the type of screening program; the age of the infant at the time of screening; and any legal action that resulted from the miss. The 76 missed cases probably represent an underascertainment of the true number, yet we believe that our data provide an overview of some of the problems associated with mass neonatal screening. There was one missed case of PKU for every 70 cases detected, and one missed case of congenital hypothyroidism for every 120 cases detected; in other words, two congenital hypothyroidism cases were missed for every 1 million infants screened. Regarding the stage of screening in which the miss occurred, 14% occurred during specimen collection, 45% during the laboratory procedures stage, 16% during follow-up, 11% were the result of biologic variation, and in 14% the stage could not be identified. We conclude that neonatal screening programs have been highly successful but that there may be additional safeguards to be developed, tested, and implemented when practical.


Versatility and addiction in gaming: the number of video-game genres played is associated with pathological gaming in male adolescents.
This study tested the predictive power of gaming versatility (i.e., the number of video game genres engaged in) on game addiction in male adolescents, controlling for time spent on gaming. Participants were 701 male adolescents attending high school (Mage=15.6 years). Analyses showed that pathological gaming was predicted not only by higher time spent on gaming, but also by participation in a greater number of video game genres. Specifically, the wider the array of video game genres played, the higher were the negative consequences caused by gaming. Findings show that versatility can be considered as one of the behavioral risk factors related to gaming addiction, which may be characterized by a composite and diversified experience with video games. This study suggests that educational efforts designed to prevent gaming addiction among youth may also be focused on adolescents' engagement in different video games.


Twenty-seven years of liver transplantation at Addenbrooke's Hospital, Cambridge.
Twenty-seven years ago, liver transplantation at Addenbrooke's Hospital was a very experimental procedure. It is now recognized as an accepted mode of treatment for end-stage liver failure. In the coming year, over 600 liver transplants will take place in the United Kingdom. All western European countries now have an active liver transplantation program and more than 17,000 liver transplants have already been performed. As results improve and the procedure becomes more readily and widely accepted, the donor shortage is likely to get worse, and will be only partially met by the introduction of split livers and living-related donors.


Comparative efficacy of two microdoses of a potentized homeopathic drug, arsenicum album, to ameliorate toxicity induced by repeated sublethal injections of arsenic trioxide in mice.
To evaluate the efficacy of 2 potentized homeopathic remedies of Arsenicum Album (Ars Alb)--6C and 30C--in combating chronic arsenic toxicity induced by repeated sublethal injections in mice (Mus musculus). Mice were randomized and divided into sets: (1) normal (control 1); (2) normal + succussed alcohol (control 2); (3) As(2)O(3) (0.016%) injected at 1 ml/100 g body weight every 7 days (treated); (4) As(2)O(3) injected + succussed alcohol (positive control); (5) As(2)O(3) injected + Ars Alb 6C (drug-fed); (6) As(2)O(3) injected + Ars Alb 30C (drug-fed). Cytogenetical endpoints like chromosome aberrations, micronuclei, mitotic index, sperm head abnormality and biochemical protocols like acid and alkaline phosphatases, aspartate and alanine aminotransferases, reduced glutathione, lipid peroxidation, catalase and succinate dehydrogenase were studied at 30, 60, 90 and 120 days. Compared to controls, chromosome aberrations, micronuclei, sperm head abnormality frequencies and activities of acid and alkaline phosphatases, aspartate and alanine aminotransferases and lipid peroxidation were reduced in both drug-fed series, while mitotic index and activities of glutathione, catalase and succinate dehydrogenase were increased. Ars Alb 30C showed marginally better efficacy than Ars Alb 6C. Both remedies indicated potentials of use against arsenic intoxication.


Epidemiological burden of postmenopausal osteoporosis in the UK from 2010 to 2021: estimations from a disease model.
This article describes the adaptation of a model estimating the burden of postmenopausal osteoporosis (PMO) to the UK. The purpose of this study was to estimate the present and future epidemiology of PMO in the UK. For each year of the study, the 'incident cohort' (women experiencing a first osteoporotic fracture) was identified and run through a Markov model using 1-year cycles until 2020. Health states were based on the number of fractures and death. Fracture by site was tracked for each health state. Transition probabilities reflected fracture site-specific risk of death and subsequent fractures. Assuming that the rate of incident fractures by age is constant over time, the model estimated the total number of PMO fractures at 224,219 in 2010, including 51,927 hip and 30,994 clinical vertebral fractures. These estimates were predicted to increase by 17.2%, 16.6% and 17.9%, respectively, by 2020. The number of postmenopausal women living with osteoporosis was predicted to increase from 1.8 million in 2010 to 2.1 million in 2020 (+16.5%). A sensitivity analysis demonstrated that the estimated number of fractures is most sensitive to the assumption made on the trends in the rate of incidence. The PMO disease model, first developed for Sweden, was adapted to the UK. Due to demographic changes, the burden of osteoporosis is expected to increase by almost a fifth by 2020. Due to the lack of country-specific data, these results rely on several assumptions regarding the incidence of non-hip fractures, trends in BMD and rate of incidence over time.


Direct estimation of inhomogeneous Markov interval models of spike trains.
A necessary ingredient for a quantitative theory of neural coding is appropriate "spike kinematics": a precise description of spike trains. While summarizing experiments by complete spike time collections is clearly inefficient and probably unnecessary, the most common probabilistic model used in neurophysiology, the inhomogeneous Poisson process, often seems too crude. Recently a more general model, the inhomogeneous Markov interval model (Berry & Meister, 1998 ; Kass & Ventura, 2001 ), was considered, which takes into account both the current experimental time and the time from the last spike. Several techniques were proposed to estimate the parameters of these models from data. Here we propose a direct method of estimation that is easy to implement, fast, and conceptually simple. The method is illustrated with an analysis of sample data from the cat's superior colliculus.


Growth phase-dependent production of a cell wall-associated elastinolytic cysteine proteinase by Staphylococcus epidermidis.
Staphylococcus epidermidis, a Gram-positive, coagulase-negative bacterium is a predominant inhabitant of human skin and mucous membranes. Recently, however, it has become one of the most important agents of hospital-acquired bacteriemia, as it has been found to be responsible for surgical wound infections developed in individuals with indwelling catheters or prosthetic devices, as well as in immunosupressed or neutropenic patients. Despite their medical significance, little is known about proteolytic enzymes of S. epidermidis and their possible contribution to the bacterium's pathogenicity; however, it is likely that they function as virulence factors in a manner similar to that proposed for the proteases of Staphylococcus aureus. Here we describe the purification of a cell wall-associated cysteine protease from S. epidermidis, its biochemical properties and specificity. A homology search using N-terminal sequence data revealed similarity to staphopain A (ScpA) and staphopain B (SspB), cysteine proteases from S. aureus. Moreover, the gene encoding S. epidermidis cysteine protease (Ecp) and a downstream gene coding for a putative inhibitor of the protease form an operon structure which resembles that of staphopain A in S. aureus. The active cysteine protease was detected on the bacterial cell surface as well as in the culture media and is apparently produced in a growth phase-dependent manner, with initial expression occurring in the mid-logarithmic phase. This enzyme, with elastinolytic properties, as well as the ability to cleave alpha1PI, fibrinogen and fibronectin, may possibly contribute to the invasiveness and pathogenic potential of S. epidermidis.


Pediatric liver transplantation: 10 years of experience at a single center in Brazil.
To evaluate the first 10 years' experience of the liver transplantation department at the Alfa Institute, Hospital das Clínicas da Universidade Federal de Minas Gerais, Brazil. A descriptive study, based on a retrospective analysis of 84 children and adolescents enrolled on a liver transplantation waiting list, from March 1995 to January 2006, based on the following variables: age, etiology of underlying liver disease, Child-Pugh, Malatack, model for end-stage liver disease (MELD) and pediatric end-stage liver disease (PELD) scores, time on waiting list, complications and survival after the procedure. Forty children had 42 liver transplants. Twenty six died while on the waiting list. Biliary atresia was the most frequent indication for transplant. The median age was 6.6 years (ranging from 1.9 to 16.8 years). Post liver transplant mortality was 32.5% (13 of 40 children). The median time on the liver transplant waiting list was 291 days. Complications related to the graft occurred in 24 of 42 transplants (57.1%), including vascular complications (30.8%), with thrombosis of the hepatic artery being the most frequent (16.6%); acute rejection occurred in 16.6%. The overall results are similar to what can be found in the literature with relation to indications and post-transplant survival. However, there were elevated rates of complications unrelated to the graft and of complications involving the hepatic artery.