Point-of-contact Interactive Record Linkage (PIRL): A software tool to prospectively link demographic surveillance and health facility data.
Linking a health and demographic surveillance system (HDSS) to data from a health facility that serves the HDSS population generates a research infrastructure for directly observed data on access to and utilization of health facility services. Many HDSS sites, however, are in areas that lack unique national identifiers or suffer from data quality issues, such as incomplete records, spelling errors, and name and residence changes, all of which complicate record linkage approaches when applied retrospectively. We developed Point-of-contact Interactive Record Linkage (PIRL) software that is used to prospectively link health records from a local health facility to an HDSS in rural Tanzania. This prospective approach to record linkage is carried out in the presence of the individual whose records are being linked, which has the advantage that any uncertainty surrounding their identity can be resolved during a brief interaction, whereby extraneous information (e.g., household membership) can be referred to as an additional criterion to adjudicate between multiple potential matches. Our software uses a probabilistic record linkage algorithm based on the Fellegi-Sunter model to search and rank potential matches in the HDSS data source. Key advantages of this software are its ability to perform multiple searches for the same individual and save patient-specific notes that are retrieved during subsequent clinic visits. A search on the HDSS database (n=110,000) takes less than 15 seconds to complete. Excluding time spent obtaining written consent, the median duration of time we spend with each patient is six minutes. In this setting, a purely automated retrospective approach to record linkage would have only correctly identified about half of the true matches and resulted in high linkage errors; therefore highlighting immediate benefit of conducting interactive record linkage using the PIRL software.


Mechanistic roles of autophagy in hematopoietic differentiation.
Autophagy is increasingly recognized for its active role in development and differentiation. In particular, its role in the differentiation of hematopoietic cells has been extensively studied, likely because blood cells are accessible, easy to identify and purify, and their progenitor tree is well defined. This review aims to discuss the mechanisms by which autophagy impacts on differentiation, using hematopoietic cell types as examples. Autophagy's roles include the remodeling during terminal differentiation, the maintenance of a long-lived cell type, and the regulation of the balance between self-renewal and quiescence in stem-like cells. We discuss and compare the mechanistic roles of autophagy, such as prevention of apoptosis, supply of energy metabolites and metabolic adaption, and selective degradation of organelles and of regulatory factors.


[Method for the study of pulmonary function in laryngectomized patients].
The aim of the study is to develop a instrumental method capable to achieve pulmonary air function test (spirometry) by assembling the spirometer mouthpiece to the laryngectomized patient's stoma. Our study was carried out in 33 laryngectomized patients (all male). The spirometer tests were done with a Datospir 92 by Sibelmed equipment, which consists of a dry Fleish pneumotacographer with flow and volume chart register. We have made a stoma-spirometer adapter with a cardboard tube, an adhesive and silicone disc. The whole sample achieved excellent outcome with the stoma-spirometer adapter. Nor air leak neither high resistence were measured while spirometry was performed. We consider that the facts studied may enable us to add, pragmatically, new resources to the more effective understanding of the respiratory handicap in the laryngectomized population.


The Promise of the Dual Prevention Pill: A Framework for Development and Introduction.
Women of reproductive age need multipurpose prevention technology (MPT) products to address two overlapping health risks: unintended pregnancy and HIV. Currently, condoms are the only available MPT, however male condoms are not within the control of a woman, and the use of female condoms has been limited by low acceptability and cost. Oral pre-exposure prophylaxis (PrEP) is highly effective for HIV prevention, yet uptake and adherence among women have been low to date. Women globally need more options for HIV and pregnancy prevention. Several MPTs for simultaneous HIV and pregnancy prevention are in various stages of development and clinical testing, although most are many years away from market launch. A dual prevention pill (DPP), a daily oral pill combining oral contraceptives and PrEP, both of which are licensed, approved products in many low- and middle-income countries (LMIC), is likely to be the fastest route to getting an MPT product into the hands of women. The DPP is one option that could enhance method choice, particularly for women who are already using oral contraceptives. By leveraging the oral contraceptive market and reaching women currently using condoms or with an unmet need for contraception, the DPP has the potential to increase the uptake of PrEP. The successful rollout of the DPP will require careful consideration of user-, provider-, and product-centered factors during product development and introduction. Early attention to these interrelated factors can help ensure that the DPP has the ideal characteristics for maximum product acceptability, that effective and quality services are designed and implemented, and that users can make informed choices, demand the product, and use it effectively. The proposed framework outlines key considerations for the effective development and introduction of the DPP, which could also facilitate integration models for future MPTs.


Motor recovery following capsular stroke. Role of descending pathways from multiple motor areas.
The functional anatomy of motor recovery was studied by assessing motor function quantitatively in 23 patients following capsular or striatocapsular stroke. While selective basal ganglia lesions (caudate and/or putamen exclusively) did not affect voluntary movements of the extremities, lesions of the anterior (plus caudate/putamen) or posterior limb of the internal capsule led to an initially severe motor impairment followed by excellent recovery, hand function included. In contrast, lesions of the posterior limb of the internal capsule in combination with damage to lateral thalamus compromised motor outcome. In experimental tracing of the topography of the internal capsule in macaque monkeys, we found axons of primary motor cortex passing through the middle third of the posterior limb of the internal capsule. Axons of premotor cortex (dorsolateral and post-arcuate area 6) passed through the capsular genu, and those of supplementary motor area (mesial area 6) through the anterior limb. Small capsular lesion can therefore disrupt the output of functionally and anatomically distinct motor areas selectively. The clinically similar motor deficits with a similar course of functional restitution following disruption of these different descending motor pathways indicate a parallel operation of cortical motor areas. They may have the further capability of substituting each other functionally in the process of recovery from hemiparesis.


Drug targeting using thermally responsive polymers and local hyperthermia.
We report a new thermal targeting method in which a thermally responsive drug carrier selectively accumulates in a solid tumor that is maintained above physiological temperature by externally applied, focused hyperthermia. We synthesized two thermally responsive polymers that were designed to exhibit a lower critical solution temperature (LCST) transition slightly above physiological temperature: (1) a genetically engineered elastin-like polypeptide (ELP) and (2) a copolymer of N-isopropylacrylamide (NIPAAm) and acrylamide (AAm). The delivery of systemically injected polymer-rhodamine conjugates to solid tumors was investigated by in vivo fluorescence video microscopy of ovarian tumors implanted in dorsal skin fold window chambers in nude mice, with and without local hyperthermia. When tumors were heated to 42 degrees C, the accumulation of a thermally responsive ELP with a LCST of 40 degrees C was approximately twofold greater than the concentration of the same polymer in tumors that were not heated. Similar results were also obtained for a thermally responsive poly(NIPAAM-co-AAm), though the enhanced accumulation of this carrier in heated tumors was lower than that observed for the thermally responsive ELP. These results suggest that enhanced delivery of drugs to solid tumors can be achieved by conjugation to thermally responsive polymers combined with local heating of tumors.


MASTR-MS: a web-based collaborative laboratory information management system (LIMS) for metabolomics.
An increasing number of research laboratories and core analytical facilities around the world are developing high throughput metabolomic analytical and data processing pipelines that are capable of handling hundreds to thousands of individual samples per year, often over multiple projects, collaborations and sample types. At present, there are no Laboratory Information Management Systems (LIMS) that are specifically tailored for metabolomics laboratories that are capable of tracking samples and associated metadata from the beginning to the end of an experiment, including data processing and archiving, and which are also suitable for use in large institutional core facilities or multi-laboratory consortia as well as single laboratory environments. Here we present MASTR-MS, a downloadable and installable LIMS solution that can be deployed either within a single laboratory or used to link workflows across a multisite network. It comprises a Node Management System that can be used to link and manage projects across one or multiple collaborating laboratories; a User Management System which defines different user groups and privileges of users; a Quote Management System where client quotes are managed; a Project Management System in which metadata is stored and all aspects of project management, including experimental setup, sample tracking and instrument analysis, are defined, and a Data Management System that allows the automatic capture and storage of raw and processed data from the analytical instruments to the LIMS. MASTR-MS is a comprehensive LIMS solution specifically designed for metabolomics. It captures the entire lifecycle of a sample starting from project and experiment design to sample analysis, data capture and storage. It acts as an electronic notebook, facilitating project management within a single laboratory or a multi-node collaborative environment. This software is being developed in close consultation with members of the metabolomics research community. It is freely available under the GNU GPL v3 licence and can be accessed from, https://muccg.github.io/mastr-ms/.


Identification of fruit yield loci controlling the salt tolerance conferred by solanum rootstocks.
The rootstock effect on the fruit yield of a grafted tomato variety was genetically analyzed under salinity using as rootstock two populations of F(9) lines developed from a salt sensitive genotype of Solanum lycopersicum var. cerasiforme, as female parent, and two salt tolerant lines, as male parents, from S. pimpinellifolium, the P population (123 lines), and S. cheesmaniae, the C population (100 lines). There were rootstock lines from the two populations (up to 65% in the P population) that raised the fruit yield of the commercial hybrid under saline conditions. It is shown that this salt tolerance rootstock effect is a heritable trait (h (2) near 0.3), governed by at least eight QTLs. The most relevant component was the number of fruits. Thus most detected QTLs correspond to this component. In general, QTL gene effects are medium-sized, with contributions from 8.5 up to 15.9% at most, and the advantageous allele comes from the wild, salt tolerant species. Only two fruit yield QTLs on chromosomes P9 and C11 might correspond to fruit yield QTLs of the non-grafted lines indicating their root system dependence. A fruit yield QTL on chromosome 3 is acting epistatically in both populations. The epistatic interactions found were dominant and they were unveiled using the associated marker as cofactor in the composite interval mapping methodology. Therefore, an efficient and profitable utilization of wild germplasm can be carried out through the improvement of rootstocks that confer salt tolerance in terms of fruit yield to the grafted variety.


Transactional processes in child disruptive behavior and maternal depression: a longitudinal study from early childhood to adolescence.
Although much has been written about the utility of applying Sameroff and Chandler's transactional perspective to the study of child psychopathology, relatively few researchers have used such an approach to trace the emergence of child problem behavior from infancy to adolescence. Using a sample of 289 male toddlers from predominantly low-income families, the current study examined associations between various forms of early child disruptive behavior, subsequent trajectories of maternal depressive symptoms over the course of 8 years, and adolescent problem behavior. Results indicated that early child noncompliance was the most robust predictor of more chronic and elevated trajectories of maternal depression, which in turn discriminated teacher and youth reports of adolescent antisocial behavior but not internalizing symptoms. The findings were consistent with transactional perspectives of developmental psychopathology that have emphasized the dynamic interplay between child and parent characteristics.


THE EFFECT OF TAX PREFERENCES ON HEALTH SPENDING.
In this paper, we estimate the effect of the tax preference for health insurance on health care spending using data from the Medical Expenditure Panel Surveys from 1996-2005. We use the fact that Social Security taxes are only levied on earnings below a statutory threshold to identify the impact of the tax preference. Because employer-sponsored health insurance premiums are excluded from Social Security payroll taxes, workers who earn just below the Social Security tax threshold receive a larger tax preference for health insurance than workers who earn just above it. We find a significant effect of the tax preference, consistent with previous research.


Isolation and Polyphasic Characterization of Desulfuromonas versatilis sp. Nov., an Electrogenic Bacteria Capable of Versatile Metabolism Isolated from a Graphene Oxide-Reducing Enrichment Culture.
In this study, a novel electrogenic bacterium denoted as strain NIT-T3 of the genus Desulfuromonas was isolated from a graphene-oxide-reducing enrichment culture that was originally obtained from a mixture of seawater and coastal sand. Strain NIT-T3 utilized hydrogen and various organic acids as electron donors and exhibited respiration using electrodes, ferric iron, nitrate, and elemental sulfur. The strain contained C16:1ω7c, C16:0, and C15:0 as major fatty acids and MK-8, 9, and 7 as the major respiratory quinones. Strain NIT-T3 contained four 16S rRNA genes and showed 95.7% similarity to Desulfuromonasmichiganensis BB1T, the closest relative. The genome was 4.7 Mbp in size and encoded 76 putative c-type cytochromes, which included 6 unique c-type cytochromes (<40% identity) compared to those in the database. Based on the physiological and genetic uniqueness, and wide metabolic capability, strain NIT-T3 is proposed as a type strain of 'Desulfuromonas versatilis' sp. nov.


Physical activity and cancer survivorship.
There has been an increase in the cancer survivor population in the United States over the past several decades primarily due to improvements in early detection of first malignancies and effective treatment modalities. A wealth of evidence has demonstrated that regular physical activity is associated with a lower risk of death, all-cause mortality, cancer recurrence, and several chronic diseases, including type 2 diabetes and cardiovascular disease, common comorbid conditions in people who have survived cancer. Physical activity also is a central component of weight management. This review summarizes the current physical activity recommendations and the evidence linking physical activity to improvements in weight management, physiological effects, and psychological health outcomes for cancer survivors. The available literature suggests physical activity is safe and is positively associated with weight management, cardiorespiratory fitness, muscular strength and endurance, quality of life, fatigue, and other psychosocial factors in cancer survivors. Yet relationships related to specific cancer diagnoses, treatments, and underlying cardiometabolic mechanisms associated with survival have not been thoroughly examined in randomized controlled trials. Furthermore, factors that influence adherence to physical activity behaviors must be identified to develop effective exercise programs. The use of objective measures of physical activity and the standardization of reporting outcome measures within intervention trials are needed to complement this effort. Healthcare providers should consider individual differences among cancer survivors and tailor physical activity programs to meet the individual needs of the patient to assist in the adoption and maintenance of a physically active lifestyle.


Management of chronic nonmalignant pain: a needs assessment in an internal medicine resident continuity clinic.
The authors hypothesized that Internal Medicine (IM) residents experience a lack of preparation, confidence, and reward when managing patients with chronic nonmalignant pain (CNMP) in their continuity clinic and that they exhibit deficiencies in CNMP management practices, particularly when opioids are prescribed. As part of a quality improvement project in the IM resident continuity clinic, the authors performed a needs assessment through a self-administered resident questionnaire and a retrospective chart review. Fifty-seven percent of respondents rated their CNMP preparation as "fair" or "poor," 89 percent reported that their experience was "much less" or "somewhat less" rewarding than managing patients with other chronic conditions, and 58 percent reported that CNMP management "negatively" or "very negatively" affected their view of primary care as a career. Twenty-eight charts of patients receiving opioids during a 1-year study period were reviewed. Although residents were likely to document pain diagnoses (93 percent) and pain scores (82 percent) as well as utilize medication agreements (82 percent), they were less likely to document illicit substance use (39 percent), document legal history (32 percent), or obtain prior medical records (39 percent). Few urine drug screens were ordered (18 percent) and 25 percent of patients had fewer than four face-to-face visits during the year. The questionnaire indicated that IM residents lack preparation in managing CNMP, which results in lack of confidence and reward. The chart review revealed management practice deficiencies in risk assessment and prescription drug misuse monitoring. As a result, the authors have implemented curricular interventions, integrated a pain clinic within the continuity clinic, optimized residency program clinic scheduling, and developed tools for consistency in management practices.


Role of inflammation in atrial fibrillation pathophysiology and management.
Atrial fibrillation (AF) is the most common clinically relevant arrhythmia, but the methods available for treating AF and its complications (of which the most important is thrombogenesis), as well as for assessing AF risk and underlying pathophysiology, are largely limited. Emerging evidence suggests a significant role of inflammation in the pathogenesis of AF. That evidence includes elevated serum levels of inflammatory biomarkers in AF subjects, the expression of inflammatory markers in cardiac tissues of AF patients and animal models of AF, and beneficial effects of anti-inflammatory drugs in experimental AF paradigms. Inflammation is suggested to be linked to various pathological processes, such as oxidative stress, apoptosis, and fibrosis, that promote AF substrate formation. Inflammation has also been associated with endothelial dysfunction, platelet activation, and coagulation cascade activation, leading to thrombogenesis. Thus, inflammation may contribute to both the occurrence/maintenance of AF and its thromboembolic complications. Here, we review the evidence for a role of inflammation and inflammatory biomarkers in the risk management and treatment of AF. We also summarize the current knowledge of inflammation-dependent cellular and molecular mechanisms in AF pathophysiology and their potential as therapeutic targets.


Treatment of cryptosporidiosis in immunocompromised individuals: systematic review and meta-analysis.
Cryptosporidium is a common cause of gastroenteritis and is associated with severe life-threatening illness among immunocompromised individuals. This review aimed to assess the efficacy of interventions for the treatment and prevention of cryptosporidiosis among immunocompromised patients. A search of Medline, Embase and other electronic databases was carried out up to August 2005. Two reviewers independently extracted data and assessed study quality. The relative risk for each intervention was calculated. Seven trials involving 169 participants were included. Nitazoxanide and paramomycin were associated with a relative risk (RR) of reduction in the duration and frequency of diarrhoea of 0.83 [95% confidence interval (CI) 0.36, 1.94] and 0.74 (95% CI 0.42, 1.31), respectively, showing no evidence of effectiveness. Nitazoxanide led to significant evidence of oocyst clearance compared with placebo with a RR of 0.52 (95% CI 0.30, 0.91). The effect was not significant for HIV-seropositive participants (RR 0.71, 95% CI 0.36, 1.37). HIV-seronegative participants on nitazoxanide had a significantly higher relative risk of achieving parasitological clearance of 0.26 (95% CI 0.09, 0.80) based on a single study. No other intervention was associated with either a reduction in diarrhoea, mortality or a significant parasitological response. This review confirms the absence of evidence for effective agents in the management of cryptosporidiosis. The results indicate that nitaxozanide reduces load of parasites and may be useful in immunocompetent individuals. The absence of effective therapy highlights the importance of preventive interventions in this group of patients.


Use of a corneal impression membrane and PCR for the detection of herpes simplex virus type-1.
Purpose. To investigate the use of a corneal impression membrane (CIM) for the detection of herpes simplex virus type 1 (HSV-1) in suspected herpes simplex keratitis (HSK).Methodology. In the laboratory study, swabs and CIMs made from polytetrafluoroethylene were spiked with different concentrations of HSV-1. DNA was extracted and real-time PCR undertaken using two sets of primers. In the clinical study, consecutive patients presenting with suspected HSK were included. For each patient, samples were collected from corneal lesions with a swab and a CIM in random order. Clinical details were collected using a standardized clinical form and patients were categorized into probable, presumed and possible HSK.Results. There was no difference in the performance of both primer sets for all HSV-1 dilutions (P=0.83) using a CIM or between a CIM and a swab (P=0.18). In total, 110 patients were included. Overall, 73 patients (66.4 %) had probable, 20 patients (18.2 %) presumed and 17 patients (15.5 %) possible HSV-1 keratitis. The HSV-1 detection rate was significantly higher using a CIM (40/110, 36.4 %) than a swab (28/110, 25.5 %) (P=0.004). In the probable HSV keratitis group, the detection rate using a CIM was 43.8 % compared to 27.4 % for a swab (P=0.004). The cycle threshold values obtained for the conjunctival swabs were higher than those obtained for the CIMs (P<0.001).Conclusions. In suspected HSK, a CIM is a useful alternative to a swab and more likely to detect the presence of HSV-1.


Long-term pathological consequences of prenatal infection: beyond brain disorders.
Prenatal immunological adversities such as maternal infection have been widely acknowledged to contribute to an increased risk of neurodevelopmental brain disorders. In recent years, epidemiological and experimental evidence has accumulated to suggest that prenatal exposure to immune challenges can also negatively affect various physiological and metabolic functions beyond those typically associated with primary defects in CNS development. These peripheral changes include excessive accumulation of adipose tissue and increased body weight, impaired glycemic regulation and insulin resistance, altered myeloid lineage development, increased gut permeability, hyperpurinergia, and changes in microbiota composition. Experimental work in animal models further suggests that at least some of these peripheral abnormalities could directly contribute to CNS dysfunctions, so that normalization of peripheral pathologies could lead to an amelioration of behavioral deficits. Hence, seemingly unrelated central and peripheral effects of prenatal infection could represent interrelated pathological entities that emerge in response to a common developmental stressor. Targeting peripheral abnormalities may thus represent a valuable strategy to improve the wide spectrum of behavioral abnormalities that can emerge in subjects with prenatal infection histories.


Neuroanatomical and functional analysis of neural tube formation in notochordless Xenopus embryos; laterality of the ventral spinal cord is lost.
Notochordless Xenopus embryos were produced by u.v. irradiation of the uncleaved fertilized egg. The spinal cords were examined using intermediate filament staining for glial cells, retrograde HRP staining for neuronal morphology and an anti-glycinergic antibody to reveal commissural cells and axons. The floorplate cells of the normal cord appear to be absent and their position along the ventral midline of the cord is occupied by motor neurones, Kolmer-Agduhr cells, radial glial cells and a ventrally placed marginal zone containing the longitudinal axons. Motor neurone number is reduced to 15% of control values, and the sensory extramedullary cell number is increased twentyfold. Commissural axons are still able to cross the ventral cord but do so at abnormal angles and some commissural axons continue to grow circumferentially up the contralateral side of the cord rather than turning to grow longitudinally. Extracellular electrophysiological recordings from motor axons reveal that the normal alternation of locomotor activity on the left and right side of the embryo is lost in notochordless animals. These results suggest that the notochord and/or the normal floor plate structure are important for the development of the laterality of spinal cord connections and may influence motor neurone proliferation or differentiation.


A Global Mitigation Hierarchy for Nature Conservation.
Efforts to conserve biodiversity comprise a patchwork of international goals, national-level plans, and local interventions that, overall, are failing. We discuss the potential utility of applying the mitigation hierarchy, widely used during economic development activities, to all negative human impacts on biodiversity. Evaluating all biodiversity losses and gains through the mitigation hierarchy could help prioritize consideration of conservation goals and drive the empirical evaluation of conservation investments through the explicit consideration of counterfactual trends and ecosystem dynamics across scales. We explore the challenges in using this framework to achieve global conservation goals, including operationalization and monitoring and compliance, and we discuss solutions and research priorities. The mitigation hierarchy's conceptual power and ability to clarify thinking could provide the step change needed to integrate the multiple elements of conservation goals and interventions in order to achieve successful biodiversity outcomes.


The early diastolic myocardial velocity: a marker of increased risk in patients with coronary heart disease.
Tissue Doppler imaging (TDI) is a promising echocardiographic modality allowing quantification of myocardial performance. However, the prognostic potential of TDI in patients with acute myocardial infarction (AMI) is not yet investigated. We sought to explore the ability of TDI in identifying patients at risk for new cardiovascular events after AMI. One hundred and nineteen patients with AMI were recruited prospectively (mean age 61 years; range 32-81 years of age). Patients with diabetes mellitus (DM) were excluded. Echocardiography was performed 3-12 months after AMI. Two-dimensional (2-D) and TDI variables were recorded. The patients were followed during a mean period of 4·6 years (range 1-8 years). The primary end-point was defined as any of the following: death from any cause, non-fatal reinfarction or stroke, unstable angina pectoris, congestive heart failure requiring hospitalization and coronary revascularization procedure. Thirty patients had some form of cardiovascular events during follow-up. Seven patients had cardiovascular death, 13 patients had reinfarction and four patients had a stroke. New angina or unstable angina was recorded in 21 patients. Of these patients, 13 underwent percutaneous coronary angioplasty (PCI) or coronary artery bypass grafting (CABG). The early diastolic myocardial velocity (E(m)) emerged as the only echocardiographic variable that offered a clear differentiation between patients that presented with new cardiovascular (CV) events as compared to the corresponding group without any CV events at follow-up (P<0·05). In multivariate statistical analysis and after adjustment for age, sex, total cholesterol, body mass index (BMI) and other baseline characteristics, Em remained as independent predictors of CV events (HR, 1·18, 95% CI, 1·02-1·36; P<0·05). However, none of the investigated variables evolved as an independent predictor of cardiovascular morbidity and mortality. E(m) appears to be a sensitive echocardiographic index in identifying non-diabetic patients with AMI at risk of new cardiovascular events.


Encapsulating metal nanoparticles inside carbon nanoflakes: a stable absorbent designed from free-standing sandwiched composites.
Combination of carbon and metal materials is a good strategy to develop lightweight microwave absorbents. Taking advantage of the encapsulation property of carbon nanoflakes, a stable microwave absorber could be achieved. In this study, we managed to achieve sandwich-shaped Ni/C nanoflakes via a simple route involving a one-step hydrothermal method towards Ni(OH)2 nanosheets, followed by a calcination procedure. These sandwiched carbon nanoflakes not only increased the interfacial polarization, but also resulted in the absolute stability of metal nanoparticles. Considering the higher possibility of some support plates to be wrinkled, which would bring about imperfect sandwiched structures, the free-standing sandwiched composites were in a relatively good shape. Furthermore, the intensive conductive loss could be highly responsible for better microwave absorption properties by adjusting the carbonization temperature. The minimum reflection loss (RL) value of Ni/C composites that were obtained with a 25% loading filler ratio could reach -24.3 dB with a matching thickness of only 1.5 mm. Moreover, the effective microwave absorption bandwidth (<-10 dB) could reach to 4.0 GHz with thicknesses of both 1.3 mm and 1.4 mm. Therefore, the sandwich-shaped Ni/C nanoflakes could be exploited as an effective and lightweight microwave absorber.


Evaluation of a novel liquid embolic agent (precipitating hydrophobic injectable liquid (PHIL)) in an animal endovascular embolization model.
The choice of the embolic agent and the embolization technique can have a significant impact on the success of endovascular embolization. To evaluate a novel iodinated copolymer-based liquid embolic agent (precipitating hydrophobic injectable liquid (PHIL)) in the porcine rete mirabile (RM), serving as an endovascular embolization model. Onyx, as an established liquid embolic agent, served as comparator. Sixteen embolization procedures were performed using PHIL (n=8) or Onyx (n=8) as liquid embolic agent. Waiting time between injections was set to 30 or 60 s (n=4 per study group). Survival time after intervention was 2 hours or 7 days. Embolization characteristics (eg, procedure times, number of injections and volume of embolic agent) and embolization extent (percentage of embolized RM in post-interventional x-ray) were assessed. Post-interventional CT and histopathological analyses were performed. Embolization characteristics and embolization extent were not significantly different for PHIL and Onyx, including subgroups (eg, embolization extent 44% vs 69% (medians); p=0.101). For PHIL, extension of the waiting time from 30 to 60 s led to a significantly higher embolization extent (24% vs 72% (medians); p=0.035). Moderate disintegration and mild inflammation of the embolized blood vessels were present for both embolic agents. PHIL is feasible for transarterial embolization in an acute and subacute endovascular embolization model. In this preliminary experimental in vivo study, embolization characteristics, embolization extent, and biocompatibility seem to be similar to those of Onyx.


Bacterial Prostatitis Enhances 2-Amino-1-Methyl-6-Phenylimidazo[4,5-b]Pyridine (PhIP)-Induced Cancer at Multiple Sites.
Dietary carcinogens, such as 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), and chronic inflammation have each been implicated as etiologic agents in prostate cancer. We hypothesized that bacterial prostatitis would accelerate PhIP-induced preinvasive lesions in the rat prostate. Male Fischer 344 rats were assigned into 4 groups: Control (untreated), PhIP (200 ppm in the diet for 20 weeks), Escherichia coli (E. coli, prostatic inoculation in week 10), or PhIP + E. coli. Study animals were monitored for a total of 52 weeks and were euthanized as necessary based on strict criteria for health status and tumor burden. Animals treated with E. coli initially developed acute and chronic inflammation in all lobes of the prostate, whereas inflammation was observed predominantly in the ventral lobe at time of death. PhIP + E. coli-treated animals exhibited a marked decrease in survival compared with PhIP-alone-treated animals as a result of an increase in the number of invasive cancers that developed at multiple sites, including the skin, small intestine, and Zymbal's gland. Despite their earlier mortality, PhIP + E. coli-treated animals developed an increased average number of precancerous lesions within the prostate compared with PhIP-treated animals, with a significantly increased Ki-67 index. Multiplexed serum cytokine analysis indicated an increase in the level of circulating IL6 and IL12 in PhIP + E. coli-treated animals. Elevated serum IL6 levels correlated with the development of precancerous lesions within the prostate. These results suggest that bacterial infections and dietary carcinogens, two conceivably preventable cancer risk factors, may synergistically promote tumorigenesis.


Heterogeneous kinetics and pharmacology of synaptic inhibition in the chick auditory brainstem.
Identification of shared features between avian and mammalian auditory brainstem circuits has provided much insight into the mechanisms underlying early auditory processing. However, previous studies have highlighted an apparent difference in inhibitory systems; synaptic inhibition is thought to be slow and GABAergic in birds but to have fast kinetics and be predominantly glycinergic in mammals. Using patch-clamp recordings in chick brainstem slices, we found that this distinction is not exclusively true. Consistent with previous work, IPSCs in nucleus magnocellularis (NM) were slow and mediated by GABA(A) receptors. However, IPSCs in nucleus laminaris (NL) and a subset of neurons in nucleus angularis (NA) had rapid time courses twofold to threefold faster than those in NM. Furthermore, we found that IPSCs in NA were mediated by both glycine and GABA(A) receptors, demonstrating for the first time a role for fast glycinergic transmission in the avian auditory brainstem. Although NM, NL, and NA have unique roles in auditory processing, the majority of inhibitory input to each nucleus arises from the same source, ipsilateral superior olivary nucleus (SON). Our results demonstrate remarkable diversity of inhibitory transmission among the avian brainstem nuclei and suggest that differential glycine and GABA(A) receptor activity tailors inhibition to the specific functional roles of NM, NL, and NA despite common SON input. We additionally observed that glycinergic/GABAergic activity in NA was usually depolarizing and could elicit spiking activity in NA neurons. Because NA projects to SON, these excitatory effects may influence the recruitment of inhibitory activity in the brainstem nuclei.


Delayed graft function is a syndrome rather than a diagnosis.
Delayed graft function is a significant prognostic indicator after renal transplantation. We hypothesized that delayed graft function is not a single entity, and different patterns of delayed graft function reflect various underlying pathological processes. An analysis of 762 renal transplants was performed, showing serum creatinine was charted serially for the first 30 days after transplant. Measurements were obtained: time on hemodialysis; time to peak creatinine; time for creatinine to half; time for creatinine to within 10% of baseline. Four patterns of delayed graft function were identified. There was no association between pattern of delayed graft function, and 1-year graft survival or serum creatinine at 1 year. Time for creatinine to > 15 days was associated with a higher creatinine level at 1 year than it was with patients with time for creatinine to half < 5 days (300.6 ± 54.3 vs 211.3 ± 26.0 μmol/L; P < .01). Patients with 1-year creatinine concentrations > 180 μmol/L had longer time on hemodialysis and time for creatinine to half than did those with 1-year creatinine concentrations ≤ 180 μmol/L (9.2 ± 1.3 μmol/L vs 7.0 ± 0.7 μmol/L; P = .03; and 11.6 ± 1.7 μmol/L vs 6.0 ± 0.4 μmol/L; P < .001). Time for creatinine to half of 6.5 days (sensitivity 67.3%; specificity 79.4%; area under the curve, 0.70) was the best predictor of a 1-year creatinine concentration ≤ 180 μmol/L. Delayed graft function is not a single entity; rather; it is the most common presentation of a heterogeneous variety of pathologies. Its rate of resolution of renal function is the best predictor of long-term graft outcome.


Operative stabilization of flail chest injuries: review of literature and fixation options.
Flail chest injuries cause significant morbidity, especially in multiply injured patients. Standard treatment is typically focused on the underlying lung injury and involves pain control and positive pressure ventilation. Several studies suggest improved short- and long-term outcomes following operative stabilization of the flail segments. Despite these studies, flail chest fixation remains a largely underutilized procedure. This article reviews the relevant literature concerning flail chest fixation and describes the different implants and techniques available for fixation. Additionally, an illustrative case example is provided for description of the surgical approach. Two prospective randomized studies, five comparative studies, and a number of case series documented benefits of operative treatment of flail chest injuries, including a decreased in ventilation duration, ICU stay, rates of pneumonia, mortality, residual chest wall deformity, and total cost of care. Historically, rib fractures have been stabilized with external plates or intramedullary implants. The use of contemporary, anatomically contoured rib plates reduced the need for intraoperative plate bending. Intramedullary rib splints allowed less-invasive fixation of posterior fractures where access for plating was limited. Operative treatment can provide substantial benefits to patients with flail chest injuries and respiratory compromise requiring mechanical ventilation. The use of anatomically contoured rib plates and intramedullary splints greatly simplifies the procedure of flail chest fixation.


Effects of floral neighborhood on seed set and degree of outbreeding in a high-alpine cushion plant.
Plants flowering together may influence each other's pollination and fecundity over a range of physical distances. Their effects on one another can be competitive, neutral, or facilitative. We manipulated the floral neighborhood of the high-alpine cushion plant Eritrichium nanum in the Swiss Alps and measured the effects of co-flowering neighbors on both the number of seeds produced and the degree of inbreeding and outbreeding in the offspring, as deduced from nuclear microsatellite markers. Seed set of E. nanum did not vary significantly with the presence or absence of two Saxifraga species growing as near neighbors, but it was higher in E. nanum cushions growing at low conspecific density than in those growing at high density. In addition, floral neighborhood had no detectable effect on the degree of selfing of E. nanum, but seeds from cushions growing at low conspecific density were more highly outbred than seeds from cushions at high density. Thus, there was no evidence of either competition or facilitation between E. nanum and Saxifraga spp. as mediated by pollinators at the spatial scale of our experimental manipulation. In contrast, the greater fecundity of E. nanum cushions at low density was consistent with reduced intraspecific competition for pollinators and might also represent a beneficial effect of highly outbred seeds as brought about by more long-distance pollinator flights under low-density conditions.


Integrative physiology: defined novel metabolic roles of osteocalcin.
The prevailing model of osteology is that bones constantly undergo a remodeling process, and that the differentiation and functions of osteoblasts are partially regulated by leptin through different central hypothalamic pathways. The finding that bone remodeling is regulated by leptin suggested possible endocrinal effects of bones on energy metabolism. Recently, a reciprocal relationship between bones and energy metabolism was determined whereby leptin influences osteoblast functions and, in turn, the osteoblast-derived protein osteocalcin influences energy metabolism. The metabolic effects of bones are caused by the release of osteocalcin into the circulation in an uncarboxylated form due to incomplete gamma-carboxylation. In this regard, the Esp gene encoding osteotesticular protein tyrosine phosphatase is particularly interesting because it may regulate gamma-carboxylation of osteocalcin. Novel metabolic roles of osteocalcin have been identified, including increased insulin secretion and sensitivity, increased energy expenditure, fat mass reduction, and mitochondrial proliferation and functional enhancement. To date, only a positive correlation between osteocalcin and energy metabolism in humans has been detected, leaving causal effects unresolved. Further research topics include: identification of the osteocalcin receptor; the nature of osteocalcin regulation in other pathways regulating metabolism; crosstalk between nutrition, osteocalcin, and energy metabolism; and potential applications in the treatment of metabolic diseases.


IGF-1 Deficiency Promotes Pathological Remodeling of Cerebral Arteries: A Potential Mechanism Contributing to the Pathogenesis of Intracerebral Hemorrhages in Aging.
Clinical and experimental studies show that age-related decline in circulating insulin-like growth factor-1 (IGF-1) levels promotes the pathogenesis of intracerebral hemorrhages, which critically contribute to the development of vascular cognitive impairment and disability in older adults. Yet, the mechanisms by which IGF-1 deficiency compromises structural integrity of the cerebral vasculature are not completely understood. To determine the role of IGF-1 deficiency in pathological remodeling of middle cerebral arteries (MCAs), we compared alterations in vascular mechanics, morphology, and remodeling-related gene expression profile in mice with liver-specific knockdown of IGF-1 (Igf1f/f + TBG-Cre-AAV8) and control mice with or without hypertension induced by angiotensin-II treatment. We found that IGF-1 deficiency resulted in thinning of the media and decreased wall-to-lumen ratio in MCAs. MCAs of control mice exhibited structural adaptation to hypertension, manifested as a significant increase in wall thickness, vascular smooth muscle cell (VSMC) hypertrophy, decreased internal diameter and up-regulation of extracellular matrix (ECM)-related genes. IGF-1 deficiency impaired hypertension-induced adaptive media hypertrophy and dysregulated ECM remodeling, decreasing elastin content and attenuating adaptive changes in ECM-related gene expression. Thus, circulating IGF-1 plays a critical role in maintenance of the structural integrity of cerebral arteries. Alterations of VSMC phenotype and pathological remodeling of the arterial wall associated with age-related IGF-1 deficiency have important translational relevance for the pathogenesis of intracerebral hemorrhages and vascular cognitive impairment in elderly hypertensive patients.


Biochemically defined HIV-1 envelope glycoprotein variant immunogens display differential binding and neutralizing specificities to the CD4-binding site.
HIV-1 gp120 binds the primary receptor CD4. Recently, a plethora of broadly neutralizing antibodies to the gp120 CD4-binding site (CD4bs) validated this region as a target for immunogen design. Here, we asked if modified HIV-1 envelope glycoproteins (Env) designed to increase CD4 recognition might improve recognition by CD4bs neutralizing antibodies and more efficiently elicit such reactivities. We also asked if CD4bs stabilization, coupled with altering the Env format (monomer to trimer or cross-clade), might better elicit neutralizing antibodies by focusing the immune response on the functionally conserved CD4bs. We produced monomeric and trimeric Envs stabilized by mutations within the gp120 CD4bs cavity (pocket-filling; PF2) or by appending heterologous trimerization motifs to soluble Env ectodomains (gp120/gp140). Recombinant glycoproteins were purified to relative homogeneity, and ligand binding properties were analyzed by ELISA, surface plasmon resonance, and isothermal titration microcalorimetry. In some formats, the PF2 substitutions increased CD4 affinity, and importantly, PF2-containing proteins were better recognized by the broadly neutralizing CD4bs mAbs, VRC01 and VRC-PG04. Based on this analysis, we immunized selected Env variants into rabbits using heterologous or homologous regimens. Analysis of the sera revealed that homologous inoculation of the PF2-containing, variable region-deleted YU2 gp120 trimers (ΔV123/PF2-GCN4) more rapidly elicited CD4bs-directed neutralizing antibodies compared with other regimens, whereas homologous trimers elicited increased neutralization potency, mapping predominantly to the gp120 third major variable region (V3). These results suggest that some engineered Env proteins may more efficiently direct responses toward the conserved CD4bs and be valuable to elicit antibodies of greater neutralizing capacity.


Amyloid fibril formation from crude protein mixtures.
Amyloid fibrils have potential as bionanomaterials. A bottleneck in their commercial use is the cost of the highly purified protein typically needed as a starting material. Thus, an understanding of the role of heterogeneity in the mixtures from which amyloid fibrils are formed may inform production of these structures from readily available impure starting materials. Insulin, a very well understood amyloid-forming protein, was modified by various reagents to explore whether amyloid fibrils could still form from a heterogeneous mixture of insulin derivatives. Aggregates were characterized by thioflavin T fluorescence and transmission electron microscopy. Using acetylation, reduction carboxymethylation, reduction pyridylethylation, trypsin digestion and chymotrypsin digestion, it was shown that amyloid fibrils can form from heterogeneous mixtures of modified insulin. The modifications changed both the rate of reaction and the yield of the final product, but led to fibrillar structures, some with interesting morphologies. Well defined, long, unbranched fibrils were observed in the crude reduced carboxymethylated insulin mixture and the crude reduced pyridylethylated insulin revealed the formation of "wavy" fibrils, compared with the straighter native insulin amyloid fibrils. Although trypsin digestion inhibited fibrils formation, chymotrypsin digestion of insulin produced a mixture of long and short fibrils under the same conditions. We conclude that amyloid fibrils may be successfully formed from heterogeneous mixtures and, further, that chemical modification may provide a simple means of manipulating protein fibril assembly for use in bionanotechnological applications, enabling some design of overall morphology in the bottom-up assembly of higher order protein structures from amyloid fibrils.


Safety and decision support in oncology.
The management of patients with cancer involves the administration of complex treatment protocols with frequent monitoring of the effects of treatment on the malignant disease as well as on the general health of the patient. The number and wide variety of protocols used in treatment trials, and the amount of clinical data generated suggest the need for computer-based support. The toxicity of many of the treatments used and the severity of the disease itself underline the safety-critical nature of all decisions made by oncologists, the physicians and surgeons who treat cancer patients. This paper presents recent work on the analysis of safety issues arising from the design and implementation of a prototype decision-support system for oncologists. It illustrates the benefits of combining both informal and formal approaches to the analysis and representation of safety, firmly based on a thorough and detailed study of the domain in cooperation with oncologists, pharmacists and medical informaticians.


Composite-dissolving microneedle patches for chemotherapy and photothermal therapy in superficial tumor treatment.
A combination of chemotherapy and photothermal therapy (PTT) has emerged as a promising strategy for cancer therapy. To ensure that the chemotherapeutic drug and photothermal agent can be simultaneously delivered to the tumor site to exert their synergistic effects, a safe and efficient delivery system is needed. Herein, we fabricated doxorubicin hydrochloride (DOX)- and indocyanine green (ICG)-loaded microneedle (MN) patches (PVP@DOX/MSN@ICG) using a two-step casting process. Mesoporous silica nanoparticles (MSNs) were used to improve the ICG stability and avoid reducing its PTT efficiency in vivo. The MN patches exhibited a good skin penetration ability, and the tips of the MN patches were dissolved by the interstitial fluid to release DOX and ICG at the tumor sites. Under 808 nm laser irradiation within 2 min, the local temperature in the tumor quickly reached 48 °C at a low power of 0.34 W cm-2. A combination of chemotherapy and PTT for PVP@DOX/MSN@ICG MN patches may maximally induce human osteosarcoma MG-63 cells in vitro. Moreover, the in vivo results showed that PVP@DOX/MSN@ICG MN patches had the best antitumor effects because of synergistic chemotherapy and PTT. Therefore, the composite-dissolving MN patch is a promising strategy for enhancing the antitumor effect of chemotherapy alone and shows the potential for the synergistic therapy of superficial tumors.


Magnetic source imaging guidance of gamma knife radiosurgery for the treatment of epilepsy.
The purpose of this study was to determine if magnetic source (MS) imaging could provide useful information in the planning and performance of gamma knife radiosurgery (GKS) for epilepsy. Magnetic source imaging of interictal epileptiform dipoles was studied in 53 epilepsy surgery candidates. All patients underwent volumetric magnetic resonance (MR) imaging. Subsequently, magnetoencephalography (MEG) was performed using single or dual 37-channel units. The MR images and MEG recordings were then coregistered to produce the MS imaging data. Magnetic source imaging epileptiform data were reviewed in a blinded fashion and spatial distributions were classified as focal, regional, multiple, scattered, or none. Postresection operative photographs were compared with MS image results to determine whether extensive or partial/no resection of the MS image focus had been accomplished. Magnetoencephalography dipoles were identified in 47 patients (89%), in 46 of whom the lesions were resected. This included 20 (80%) of 25 anterior temporal lobe (ATL) cases, and 26 (93%) of 28 extratemporal lobe (ETL) cases. Of the six patients who underwent extensive ATL resections, three (50%) were seizure free. Of 14 patients who underwent partial/no resection of the ATL, seven (50%) were seizure free. There was no clear relation between MS image spatial distribution and surgery-related outcome. Of the seven ATL cases with hippocampal atrophy, five patients (71%) were seizure free. Of 12 ETL cases (three lesional), 10 patients (83%) were seizure free. Of 14 patients who underwent partial/no ETL resections (three lesional), two (14%) were seizure free. Of five nonlesional ETL cases with focal MS image dipoles, four patients (80%) were seizure free. Of five nonlesional ETL cases with regional dipoles, three patients (60%) were seizure free. Of eight ETL cases with multiple MS image dipoles, two patients (25%) were seizure free. Spatial agreement of MS imaging and electrographic data had no apparent effect on outcome of either ATL or ETL cases. Nonlesional ETL cases with focal (and in some cases multiple or regional) epileptiform MS image dipole distributions benefit significantly from inclusion of the MS image epileptiform focus in the resections. Nonlesional ETL cases suitable for GKS may similarly benefit from including the MS image focus in the irradiated area.


Positive replication and linkage disequilibrium mapping of the chromosome 21q22.1 malaria susceptibility locus.
Four cytokine receptor genes are located on Chr21q22.11, encoding the alpha and beta subunits of the interferon-alpha receptor (IFNAR1 and IFNAR2), the beta subunit of the interleukin 10 receptor (IL10RB) and the second subunit of the interferon-gamma receptor (IFNGR2). We previously reported that two variants in IFNAR1 were associated with susceptibility to malaria in Gambians. We now present an extensive fine-scale mapping of the associated region utilizing 45 additional genetic markers obtained from public databases and by sequencing a 44 kb region in and around the IFNAR1 gene in 24 Gambian children (12 cases/12 controls). Within the IFNAR1 gene, a newly studied C --> G single-nucleotide polymorphism (IFNAR1 272354c-g) at position -576 relative to the transcription start was found to be more strongly associated with susceptibility to severe malaria. Association was observed in three populations: in Gambian (P=0.002), Kenyan (P=0.022) and Vietnamese (P=0.005) case-control studies. When all three studies were combined, using the Mantel-Haenszel test, the presence of IFNAR1 -576G was associated with a substantially elevated risk of severe malaria (N=2444, OR=1.38, 95% CI: 1.17-1.64; P=1.7 x 10(-4)). This study builds on previous work to further highlight the importance of the type-I interferon pathway in malaria susceptibility and illustrates the utility of typing SNPs within regions of high linkage disequilibrium in multiple populations to confirm initial positive associations.


Crystal structure of a self-spliced group II intron.
Group II introns are self-splicing ribozymes that catalyze their own excision from precursor transcripts and insertion into new genetic locations. Here we report the crystal structure of an intact, self-spliced group II intron from Oceanobacillus iheyensis at 3.1 angstrom resolution. An extensive network of tertiary interactions facilitates the ordered packing of intron subdomains around a ribozyme core that includes catalytic domain V. The bulge of domain V adopts an unusual helical structure that is located adjacent to a major groove triple helix (catalytic triplex). The bulge and catalytic triplex jointly coordinate two divalent metal ions in a configuration that is consistent with a two-metal ion mechanism for catalysis. Structural and functional analogies support the hypothesis that group II introns and the spliceosome share a common ancestor.


The International Universities Walking Project: development of a framework for workplace intervention using the Delphi technique.
This paper aimed to use the Delphi technique to develop a consensus framework for a multinational, workplace walking intervention. Ideas were gathered and ranked from eight recognized and emerging experts in the fields of physical activity and health, from universities in Australia, Canada, England, The Netherlands, Northern Ireland, and Spain. Members of the panel were asked to consider the key characteristics of a successful campus walking intervention. Consensus was reached by an inductive, content analytic approach, conducted through an anonymous, three-round, e-mail process. The resulting framework consisted of three interlinking themes defined as "design, implementation, and evaluation." Top-ranked subitems in these themes included the need to generate research capacity (design), to respond to group needs through different walking approaches (implementation), and to undertake physical activity assessment (evaluation). Themes were set within an underpinning domain, referred to as the "institution" and sites are currently engaging with subitems in this domain, to provide sustainable interventions that reflect the practicalities of local contexts and needs. Findings provide a unique framework for designing, implementing, and evaluating walking projects in universities and highlight the value of adopting the Delphi technique for planning international, multisite health initiatives.


Role of small-fiber afferents in pain mechanisms with implications on diagnosis and treatment.
Numerous mechanisms are implicated in the perception of pain. Although many anatomical, molecular, and functional components have been identified, a comprehensive and integrated theory of pain perception has yet to be firmly established that fits the diverse clinical experience. Acute pain involves the activation of several varieties of small primary sensory neurons, collectively termed nociceptors, which have small-caliber unmyelinated or myelinated axons (C and Adelta fibers, respectively) that innervate all body tissues. They are stimulated by noxious stimuli that activate ion channels on the endings either directly or through the release of cytokines from damaged or stressed tissues. A variety of drugs successfully treats acute pain by targeting these ion channels or cytokine interactions. Paradoxically, several chronic neuropathic pain conditions are associated with a loss of small-caliber axons and have an unpredictable and poor response to current drugs, especially at doses that do not cause severe side effects. In an attempt to further an integrated theory of pain perception, this review focuses upon the presumed role of small-caliber innervation, particularly to the epidermis and cutaneous vasculature, and the clinical manifestations, diagnosis, and treatment of pathologies of this innervation.


Dendritic cells pulsed with pancreatic cancer total tumor RNA generate specific antipancreatic cancer T cells.
RNA-based dendritic cell immunotherapy with the use of total tumor RNA provides the potential to generate a polyclonal immune response to multiple known and unknown tumor antigens without HLA restriction. Our study evaluated this approach as potential immunotherapy for patients with pancreatic cancer. Dendritic cells were generated using adherent monocytes isolated from peripheral blood of patients with pancreatic cancer and evaluated phenotypically by flow cytometry to determine whether dendritic cells could be generated from the blood of patients with pancreatic cancer. Immature dendritic cells were transfected with mRNA encoding full-length carcinoembryonic antigen (CEA) or pancreatic cancer total tumor messenger RNA, and then matured. Matured dendritic cell phenotypes were also analyzed by flow cytometry. Transfected, matured dendritic cells were used to stimulate autologous T cells, and the resultant antigen-specific effector T cells were analyzed by interferon-gamma Elispot assay. Immature dendritic cells with characteristic phenotypic markers CD40, CD80, and CD86 were successfully isolated from the blood of patients with pancreatic cancer. Incubation with maturation agents increased expression of CD80 and CD83, demonstrating the induction of a mature antigen-presenting phenotype. Dendritic cells transfected with a pancreatic cancer-associated antigen (CEA) generated antigen-specific T cells (P<0.05). Dendritic cells transfected with autologous total tumor pancreatic cancer RNA generated T cells that specifically recognized HLA-matched pancreatic cancer cell lines (P<0.05 compared to control cell lines). Dendritic cells from patients with pancreatic cancer maintain the ability to translate and process transfected RNA and serve as mature antigen-presenting cells. These RNA-transfected dendritic cells from pancreatic cancer patients successfully generate specific T cells against the pancreatic cancer-associated antigen CEA as well as T cells that specifically recognize pancreatic cancer cells. These data suggest that total tumor RNA-pulsed dendritic cells may have potential as an adjuvant immunotherapy for patients with pancreatic cancer.


Reticuline exposure to invertebrate ganglia increases endogenous morphine levels.
Given the presence of morphine, its metabolites and precursors in mammalian and invertebrate tissues, it became important to determine if exposing tissues to an opiate alkaloid precursor, reticuline, would result in increasing endogenous morphine levels. Endogenous morphine levels were determined by high pressure liquid chromatography coupled to electrochemical detection and radioimmunoassay following incubation of Mytilus edulis pedal ganglia with reticuline. Nitric oxide (NO) release was determined in real-time via an amperometric probe. Mu opiate receptor affinity for opiate alkaloid precursors was determined by a receptor displacement assay. Morphine is present in the pedal ganglia of Mytilus edulis (1.43 +/- 0.41 ng/mg +/- SEM ganglionic wet weight). Ganglia incubated with 50 ng of reticuline, a morphine precursor in plants, for 1 hour exhibited a statistical increase in their endogenous morphine levels (6.7 +/- 0.7 ng/mg tissue wet weight; P<0.01). This phenomenon is concentration dependent. The increase in ganglionic morphine levels occurs gradually over the 60 min incubation period, beginning 10 minutes post reticuline addition. We show that reticuline (10(-6) M) does not stimulate ganglionic NO release in a manner resembling that of morphine (10(-6) M), which releases NO seconds after its exposure to the ganglia and lasts for 5 minutes. With reticuline, there is a 3 minute delay, which is followed by an extended release period. Furthermore, in binding displacement experiments both reticuline and salutaridine (another morphine precursor) exhibit no binding affinity for the pedal ganglia mu opiate receptor subtype. This finding is further substantiated using the positive control of human monocytes where the mu3 opiate receptor subtype has been cloned. Taken together, we surmise that the morphine's precursors are being converted to morphine. The experiments strongly indicate that pedal ganglia can synthesize morphine from reticuline.


Polarization discrimination of coherently propagating light in turbid media.
We describe the use of degree of polarization to discriminate unscattered and weakly scattered light from multiply scattered light in an optically turbid material. We use spatially resolved measurements of the degree of polarization to compare how well linearly and circularly polarized light survives in a sample. Experiments were performed on common tissue phantoms consisting of polystyrene and Intralipid microsphere suspensions and on adipose and arterial tissue. The results indicate that polarization is maintained even after unpolarized irradiance through each sample has been extinguished by several orders of magnitude. The results also show that polarized light propagation in common tissue phantoms is distinctly different from polarized light propagation in the two tissues investigated. Further, these experiments illustrate when polarization is an effective discrimination criterion and when it is not. The potential of a polarization-based discrimination scheme to image through the biological and nonbiological samples investigated here is also discussed.


Glucose regulates the effects of leptin on hypothalamic POMC neurons.
Leptin is believed to exert its potent appetite-suppressing effects via stimulation of hypothalamic anorexigenic proopiomelanocortin (POMC)-containing neurons and inhibition of orexigenic agouti-related protein (AgRP) neurons. We show here that at 11 mM glucose leptin excites POMC cells. At 5 mM glucose, however, leptin hyperpolarizes POMC neurons and suppresses action potential firing, by producing a greater decrease in excitatory synaptic tone than inhibitory tone. These results argue that when glucose is low (5 mM or less) AgRP neurons will be more important for mediating the anorectic effects of leptin than POMC cells. However, at high glucose concentrations (11 mM), activation of POMC cells may contribute to the appetite-suppressing effects of leptin. Our data also suggest the regulation of neuropeptide efficacy as a novel function of hypothalamic glucose sensing.


Initial corrosion of amalgams in vitro.
The amounts of copper, mercury, silver or zinc released from two brands of freshly prepared, lathe cut amalgams and from one brand of dispersed phase type amalgam into artificial saliva have been measured. Samples were immersed in the solution a few minutes after the end of trituration and exposed statically for periods of up to about 4 days. The initial mercury release from such specimens could exceed the long term mercury release from old amalgams by more than two orders of magnitude. The measurements indicate that during the first day after insertion of two amalgam fillings, each with an assumed surface area of 1 cm2 and under presumably static conditions, mercury at the level of more than twice the mercury food and drink intake could be released in the oral cavity. This is, however, a situation which appears infrequently, e.g. following dental treatment once a year. The amounts of copper and zinc released initially were considerably lower than the corresponding food and drink intake values, while silver might be on the same level. The measurements were conducted using nuclear tracer techniques.


On the evolution and optimality of mood States.
Moods can be regarded as fluctuating dispositions to make positive and negative evaluations. Developing an evolutionary approach to mood as an adaptive process, we consider the structure and function of such states in guiding behavioural decisions regarding the acquisition of resources and the avoidance of harm in different circumstances. We use a drift diffusion model of decision making to consider the information required by individuals to optimise decisions between two alternatives, such as whether to approach or withdraw from a stimulus that may be life enhancing or life threatening. We show that two dimensions of variation (expectation and preparedness) are sufficient for such optimal decisions to be made. These two dispositional dimensions enable individuals to maximize the overall benefits of behavioural decisions by modulating both the choice made (e.g., approach/withdraw) and decision speed. Such a structure is compatible with circumplex models of subjectively experienced mood and core affect, and provides testable hypotheses concerning the relationships that occur between valence and arousal components of mood in differing ecological niches. The paper is therefore a useful step toward being able to predict moods (and the effect of moods) using an optimality approach.


A pilot of audio computer-assisted self-interview for youth reproductive health research in Vietnam.
Several recent adolescent health studies in Vietnam have shown low levels of premarital sex among youth compared to neighboring countries and other regions of the world. One possible explanation for these findings is that adolescents in Vietnam are less willing to reveal their true behaviors. This study aims to assess the level of reporting of sensitive behaviors/events using three methods of survey data collection: face-to-face interviewer-administered (IA), paper-and-pencil self-administered (SA) and AudioComputerAssisted Self Interview (ACASI). A randomized experiment was undertaken in Gialam, a suburb of Hanoi, among a sample of 2,394 youth ages 15 to 24 years. Respondents were randomly assigned to one of three interviewing methods, with females and males evaluated separately. ACASI showed certain advantages with regard to respondent attitudes and perceptions of sensitive topics. ACAI also revealed higher prevalence rates for sensitive and stigmatized behaviors. Among those in the pencil and paper survey group it is estimated that 12.9% of unmarried males and 3.4% of unmarried females have had premarital sex. The rate found by using ACASI is higher at 17.1% in males (95% CI: 13.5-21.4) and 4.5% in females (95% CI: 2.7-7.3). Using ACASI, unmarried males also reported higher levels of risky sexual relations. For example, 7.8% confirmed visiting sex workers compared with only 1.2% in SA group and 3.9% in IA group. Additionally, ACASI respondents reported having had more sex partners by age group, gender and marital status. When coupled with the emerging data from around the world, the present findings suggest that researchers should consider using ACASI for future studies dealing with sensitive and stigmatized topics.


Health and economic burden of HPV-related diseases in Singapore.
To assess the health and economic burden of human papillomavirus (HPV)-related diseases (cervical cancer, cervical intraepithelial neoplasia (CIN) 1/2/3, and genital warts) in Singapore over a period of 25 years beginning in 2008. Incidence-based modeling was used to estimate the incidence cases and associated economic burden, with the assumption that age-stratified incidence rates will remain the same throughout the period of 25 years. The incidence rates in 2008 were projected based on data obtained from the National Cancer Registry for cervical cancer, and from a combination of published data and hospital registry review for CIN1/2/3 and genital warts. The population growth rate was factored into the projection of incidence cases over time. Direct cost data per cervical cancer and per CIN1/2/3 case were obtained from the financial database of large local hospitals while cost data for genital warts were obtained from the National Skin Center; these costs were multiplied by the number of incidence cases to produce an aggregate estimate of the economic burden over the 25-year period (in 2008 Singapore dollars) using a 3% discount rate. The total number of incidence cases of HPV-disease over 25 years beginning in 2008 was estimated to be 60,183, including 8,078 for cervical cancer, 11,685 for CIN 2/3, 8,849 for CIN1, and 31,572 for genital warts. The estimated total direct cost was 83.2 million Singapore Dollars over 25 years: 57.6 million attributable to cervical cancer, 13.0 million to CIN2/3, 6.83 million to CIN1, and 5.70 million to genital warts. HPV-related diseases are expected to impose significant health and economic burden on the Singapore healthcare resources in the next 25 years.


Objective Behavioral Measures in Children before, during, and after the COVID-19 Lockdown in Israel.
Studies using questionnaires report that COVID-19 restrictions resulted in children spending significantly less time outdoors. This study used objective measures to assess the impact of pandemic-related restrictions on children's behavior. A total of 19 healthy 8-12-year-old boys were observed before and during social restriction periods. Of these, 11 boys were reassessed after restrictions were lifted. For each session, Actiwatches were dispensed for measures of time outdoors, activity, and sleep. Changes overall and by school status were assessed using signed-rank test and Wilcoxon rank sum tests. During restrictions, children spent significantly less time outdoors (p = 0.001), were less active (p = 0.001), and spent less time engaged in moderate-to-vigorous physical activity (p = 0.004). Sleep duration was not significantly different between sessions (p > 0.99), but bedtime and wake time shifted to a later time during restrictions (p < 0.05 for both). Time outdoors and activity returned close to pre-pandemic levels after restrictions were lifted (p > 0.05 for both). Children's behaviors significantly changed during the COVID-19 pandemic. The reduction in outdoor light exposure is of importance due to the role of light in the etiology of myopia and vitamin D production. The reduction in physical activity may have negative health effects in terms of obesity and depression, although further research is required to ascertain the long-term effects.


Redox regulation of electrophilic signaling by reactive persulfides in cardiac cells.
Maintaining a redox balance by means of precisely controlled systems that regulate production, and elimination, and metabolism of electrophilic substances (electrophiles) is essential for normal cardiovascular function. Electrophilic signaling is mainly regulated by endogenous electrophiles that are generated from reactive oxygen species, nitric oxide, and the derivative reactive species of nitric oxide during stress responses, as well as by exogenous electrophiles including compounds in foods and environmental pollutants. Among electrophiles formed endogenously, 8-nitroguanosine 3',5'-cyclic monophosphate (8-nitro-cGMP) has unique cell signaling functions, and pathways for its biosynthesis, signaling mechanism, and metabolism in cells have been clarified. Reactive persulfide species such as cysteine persulfides and polysulfides that are endogenously produced in cells are likely to be involved in 8-nitro-cGMP metabolism. These new aspects of redox biology may stimulate innovative and multidisciplinary research in cardiovascular physiology and pathophysiology. In our review, we focus on the redox-dependent regulation of electrophilic signaling via reduction and metabolism of electrophiles by reactive persulfides in cardiac cells, and we include suggestions for a new therapeutic strategy for cardiovascular disease.


Spectroscopic evidence of uranium immobilization in acidic wetlands by natural organic matter and plant roots.
Biogeochemistry of uranium in wetlands plays important roles in U immobilization in storage ponds of U mining and processing facilities but has not been well understood. The objective of this work was to study molecular mechanisms responsible for high U retention by Savannah River Site (SRS) wetland sediments under varying redox and acidic (pH = 2.6-5.8) conditions using U L3-edge X-ray absorption spectroscopy. Uranium in the SRS wetland sediments existed primarily as U(VI) bonded as a bidentate to carboxylic sites (U-C bond distance at ∼2.88 Å), rather than phenolic or other sites of natural organic matter (NOM). In microcosms simulating the SRS wetland processes, U immobilization on roots was 2 orders of magnitude higher than on the adjacent brown or more distant white sands in which U was U(VI). Uranium on the roots were both U(IV) and U(VI), which were bonded as a bidentate to carbon, but the U(VI) may also form a U phosphate mineral. After 140 days of air exposure, all U(IV) was reoxidized to U(VI) but remained as a bidentate bonding to carbon. This study demonstrated NOM and plant roots can highly immobilize U(VI) in the SRS acidic sediments, which has significant implication for the long-term stewardship of U-contaminated wetlands.


The Iws1:Spt6:CTD complex controls cotranscriptional mRNA biosynthesis and HYPB/Setd2-mediated histone H3K36 methylation.
Many steps in gene expression and mRNA biosynthesis are coupled to transcription elongation and organized through the C-terminal domain (CTD) of the large subunit of RNA polymerase II (RNAPII). We showed recently that Spt6, a transcription elongation factor and histone H3 chaperone, binds to the Ser2P CTD and recruits Iws1 and the REF1/Aly mRNA export adaptor to facilitate mRNA export. Here we show that Iws1 also recruits the HYPB/Setd2 histone methyltransferase to the RNAPII elongation complex and is required for H3K36 trimethylation (H3K36me3) across the transcribed region of the c-Myc, HIV-1, and PABPC1 genes in vivo. Interestingly, knockdown of either Iws1 or HYPB/Setd2 also enhanced H3K27me3 at the 5' end of the PABPC1 gene, and depletion of Iws1, but not HYPB/Setd2, increased histone acetylation across the coding regions at the HIV-1 and PABPC1 genes in vivo. Knockdown of HYPB/Setd2, like Iws1, induced bulk HeLa poly(A)+ mRNAs to accumulate in the nucleus. In vitro, recombinant Spt6 binds selectively to a stretch of uninterrupted consensus repeats located in the N-terminal half of the CTD and recruits Iws1. Thus Iws1 connects two distinct CTD-binding proteins, Spt6 and HYPB/Setd2, in a megacomplex that affects mRNA export as well as the histone modification state of active genes.


Diffuse Staining for Activated NOTCH1 Correlates With NOTCH1 Mutation Status and Is Associated With Worse Outcome in Adenoid Cystic Carcinoma.
NOTCH1 is frequently mutated in adenoid cystic carcinoma (ACC). To test the idea that immunohistochemical (IHC) staining can identify ACCs with NOTCH1 mutations, we performed IHC for activated NOTCH1 (NICD1) in 197 cases diagnosed as ACC from 173 patients. NICD1 staining was positive in 194 cases (98%) in 2 major patterns: subset positivity, which correlated with tubular/cribriform histology; and diffuse positivity, which correlated with a solid histology. To determine the relationship between NICD1 staining and NOTCH1 mutational status, targeted exome sequencing data were obtained on 14 diffusely NICD1-positive ACC specimens from 11 patients and 15 subset NICD1-positive ACC specimens from 15 patients. This revealed NOTCH1 gain-of-function mutations in 11 of 14 diffusely NICD1-positive ACC specimens, whereas all subset-positive tumors had wild-type NOTCH1 alleles. Notably, tumors with diffuse NICD1 positivity were associated with significantly worse outcomes (P=0.003). To determine whether NOTCH1 activation is unique among tumors included in the differential diagnosis with ACC, we performed NICD1 IHC on a cohort of diverse salivary gland and head and neck tumors. High fractions of each of these tumor types were positive for NICD1 in a subset of cells, particularly in basaloid squamous cell carcinomas; however, sequencing of basaloid squamous cell carcinomas failed to identify NOTCH1 mutations. These findings indicate that diffuse NICD1 positivity in ACC correlates with solid growth pattern, the presence of NOTCH1 gain-of-function mutations, and unfavorable outcome, and suggest that staining for NICD1 can be helpful in distinguishing ACC with solid growth patterns from other salivary gland and head and neck tumors.


Modeling Long-Term Corn Yield Response to Nitrogen Rate and Crop Rotation.
Improved prediction of optimal N fertilizer rates for corn (Zea mays L.) can reduce N losses and increase profits. We tested the ability of the Agricultural Production Systems sIMulator (APSIM) to simulate corn and soybean (Glycine max L.) yields, the economic optimum N rate (EONR) using a 16-year field-experiment dataset from central Iowa, USA that included two crop sequences (continuous corn and soybean-corn) and five N fertilizer rates (0, 67, 134, 201, and 268 kg N ha-1) applied to corn. Our objectives were to: (a) quantify model prediction accuracy before and after calibration, and report calibration steps; (b) compare crop model-based techniques in estimating optimal N rate for corn; and (c) utilize the calibrated model to explain factors causing year to year variability in yield and optimal N. Results indicated that the model simulated well long-term crop yields response to N (relative root mean square error, RRMSE of 19.6% before and 12.3% after calibration), which provided strong evidence that important soil and crop processes were accounted for in the model. The prediction of EONR was more complex and had greater uncertainty than the prediction of crop yield (RRMSE of 44.5% before and 36.6% after calibration). For long-term site mean EONR predictions, both calibrated and uncalibrated versions can be used as the 16-year mean differences in EONR's were within the historical N rate error range (40-50 kg N ha-1). However, for accurate year-by-year simulation of EONR the calibrated version should be used. Model analysis revealed that higher EONR values in years with above normal spring precipitation were caused by an exponential increase in N loss (denitrification and leaching) with precipitation. We concluded that long-term experimental data were valuable in testing and refining APSIM predictions. The model can be used as a tool to assist N management guidelines in the US Midwest and we identified five avenues on how the model can add value toward agronomic, economic, and environmental sustainability.


In vivo mapping of tissue- and subcellular-specific proteomes in Caenorhabditis elegans.
Multicellular organisms are composed of tissues that have distinct functions requiring specialized proteomes. To define the proteome of a live animal with tissue and subcellular resolution, we adapted a localized proteomics technology for use in the multicellular model organism Caenorhabditis elegans. This approach couples tissue- and location-specific expression of the enzyme ascorbate peroxidase (APX), which enables proximity-based protein labeling in vivo, and quantitative proteomics to identify tissue- and subcellular-restricted proteomes. We identified and localized more than 3000 proteins from strains of C. elegans expressing APX in either the nucleus or cytoplasm of the intestine, epidermis, body wall muscle, or pharyngeal muscle. We also identified several hundred proteins that were specifically localized to one of the four tissues analyzed or specifically localized to the cytoplasm or the nucleus. This approach resulted in the identification both of proteins with previously characterized localizations and of those not known to localize to the nucleus or cytoplasm. Further, we confirmed the tissue- and subcellular-specific localization of a subset of identified proteins using green fluorescent protein tagging and fluorescence microscopy, validating our in vivo proximity-based proteomics technique. Together, these results demonstrate a new approach that enables the tissue- and subcellular-specific identification and quantification of proteins within a live animal.


High prevalence of unusual genotypes of Toxoplasma gondii infection in pork meat samples from Erechim, Southern Brazil.
Toxoplasmosis is the most common cause of infectious uveitis in Brazil, with a higher frequency in the South of the country. We have collected samples from porcine tongue and diaphragm obtained in both large and small abattoirs and used molecular biological technique to determine the prevalence of infection and RFLP analysis to type the parasites. Seventeen out of 50 (34%) samples from the diaphragm and 33 out of 50 (66%) samples from the tongue demonstrated a positive PCR reaction for T. gondii and restriction analysis of four of the positive samples revealed that all had a type I genotype at SAG2. However, when other unlinked loci were analyzed, these strains had a type III genotype at markers BTUB, SAG3, and GRA6. One of the strains (8T) had a type II allele at SAG3, indicating it has a combination of alleles normally seen in the clonal lineages. Our sampling indicates a high prevalence of infection and suggests that unusual genotypes of T. gondii are found in Brazil even among domesticated pigs.


Double outlet right ventricle. Study of 27 cases.
Out of 1610 children's hearts with congenital malformations there were 27 specimens showing double outlet right ventricle. Cases with dextrocardia, situs inversus, or l-venticular loop were excluded. Anatomical examination was performed with particular reference to the infundibular region, the great vessels, and the ventricular septum. The commonest associated malformations were ventricular septal defect and pulmonary stenosis. Aortic stenosis was the predominant finding in those cases dying in the neonatal period. An aortic conus was associated with pulmonary stenosis, ventricular septal defect, and d-transposition, a pulmonary conus with ventricular septal defect and a double conus with stenosis of either great vessel. The anterior vessel always had a muscular conus and the posterior vessel was commonly stenotic.


Association of Fluid Resuscitation Initiation Within 30 Minutes of Severe Sepsis and Septic Shock Recognition With Reduced Mortality and Length of Stay.
We evaluate the association of intravenous fluid resuscitation initiation within 30 minutes of severe sepsis or septic shock identification in the emergency department (ED) with inhospital mortality and hospital length of stay. We also compare intravenous fluid resuscitation initiated at various times from severe sepsis or septic shock identification's association with the same outcomes. This was a review of a prospective, observational cohort of all ED severe sepsis or septic shock patients during 13 months, captured in a performance improvement database at a single, urban, tertiary care facility (90,000 ED visits/year). The primary exposure was initiation of a crystalloid bolus at 30 mL/kg within 30 minutes of severe sepsis or septic shock identification. Secondary analysis compared intravenous fluid initiated within 30, 31 to 60, or 61 to 180 minutes, or when intravenous fluid resuscitation was initiated at greater than 180 minutes or not provided. Of 1,866 subjects, 53.6% were men, 72.5% were white, mean age was 72 years (SD 16.6 years), and mean initial lactate level was 2.8 mmol/L. Eighty-six percent of subjects were administered intravenous antibiotics within 180 minutes; 1,193 (64%) had intravenous fluid initiated within 30 minutes. Mortality was lower in the within 30 minutes group (159 [13.3%] versus 123 [18.3%]; 95% confidence interval [CI] 1.4% to 8.5%), as was median hospital length of stay (6 days [95% CI 6 to 7] versus 7 days [95% CI 7 to 8]). In multivariate regression that included adjustment for age, lactate, hypotension, acute organ dysfunction, and Emergency Severity Index score, intravenous fluid within 30 minutes was associated with lower mortality (odds ratio 0.63; 95% CI 0.46 to 0.86) and 12% shorter length of stay (hazard ratio=1.14; 95% CI 1.02 to 1.27). In secondary analysis, mortality increased with later intravenous fluid resuscitation initiation: 13.3% (≤30 minutes) versus 16.0% (31 to 60 minutes) versus 16.9% (61 to 180 minutes) versus 19.7% (>180 minutes). Median hospital length of stay also increased with later intravenous fluid initiation: 6 days (95% CI 6 to 7 days) versus 7 days (95% CI 6 to 7 days) versus 7 days (95% CI 6 to 8 days) versus 8 days (95% CI 7 to 9 days). The time of intravenous fluid resuscitation initiation was associated with improved mortality and could be used as an easier obtained alternative to intravenous fluid completion time as a performance indicator in severe sepsis and septic shock management.


Predominant intracellular expression of CXCR4 and CCR5 in purified primary trophoblast cells from first trimester and term human placentae.
The aim of the present study was to define the expression of CXCR4 and CCR5 on non-cultured non-stimulated primary human trophoblast cells (TCs) immediately after their immunopurification. We have evaluated by flow cytometric analysis and immunofluorescence, highly purified primary TCs prepared from first trimester (8.2 +/- 0.3 weeks, n = 15) and term (Caesarean section, n = 10) placentae for the cell surface and intracellular expression of CXCR4 and CCR5. There was a high level of individual variability for CXCR4 and CCR5 expression between trophoblast batches. In first trimester and term placentae TCs, we found a greater number of TCs preparations expressing intracellular CXCR4 than CCR5 (P < 0.05). Both receptors were predominantly localized in the intracellular compartment of TCs, whatever if isolated from first trimester or term placentae. The functional consequences of the predominance of CXCR4 expression and of cellular addressing are briefly discussed.


Metabolism of nicotine by rat liver cytochromes P-450. Assessment utilizing monoclonal antibodies to nicotine and cotinine.
Because of the prevalence of cigarette smoking in the general population and because studies suggest that a large percentage of nicotine is metabolized to cotinine in humans, it is important to study the enzymes responsible for nicotine metabolism. The cytochromes P-450 have long been implicated in the first step in the conversion of nicotine to nicotine delta 1'(5')-iminium ion. We demonstrate here that rat liver P-450IIB1 is able to convert nicotine to cotinine in the presence of cytosol with a Km of 5-7 microM. A constitutive form of P-450 is also implicated in nicotine metabolism, while purified P-450IA1 and P-450IIC6 show no detectable activity. The lack of P-450IA1 activity substantiates work by others who also failed to observe an increase in the efficiency of nicotine metabolism to cotinine by microsomes from rats that had been pretreated with benzanthracene. This result is in contrast to work with purified rabbit liver enzymes, in which P-450IA1 exhibited low but measurable activity. Our results support the notion that nicotine metabolism to cotinine by P-450 enzymes is highly species dependent. Thus, it is unwise in some cases to extrapolate results obtained by animal model study to the possible role of specific forms of the P-450 enzymes in nicotine metabolism in humans.


Testosterone replacement therapy for the treatment of neurological and neuropsychiatric disorders.
There is an accumulating body of evidence in the literature suggesting that testosterone may be neuroprotective and therefore have therapeutic value in the treatment of neurodegenerative diseases. In this article, the potential mechanisms of action of testosterone on the central nervous system are discussed, as well as the current evidence supporting the effect of testosterone on neurological impairment. Lower endogenous testosterone levels appear to be associated with some neurological disabilities. Conversely, a therapeutic benefit of exogenous testosterone supplementation on some cognitive abilities has been shown in pilot trials involving healthy elderly men as well as patients with multiple sclerosis and Alzheimer's disease. Further study of the therapeutic potential of testosterone is warranted in neurological and neuropsychiatric diseases.


The effects of contraceptive education on method use at first intercourse.
Despite long-standing public support for sex education in the schools, it has been difficult to show concrete effects of sex education on sexual and contraceptive behavior. Data from the 1988 National Survey of Family Growth indicate that exposure to a formal contraceptive education program increases the likelihood that a teenage woman will use a contraceptive method at first intercourse. According to the results of a multivariate analysis, the odds that a young woman will use any method and the odds that she will use a condom increase by about one-third following instruction about birth control; the effect on the likelihood of pill use, however, is nonsignificant. If contraceptive education occurs in the same year that a teenager becomes sexually active, the odds of any method use and of condom use are increased by 70-80%, and the odds of pill use are more than doubled. The results also suggest that with greater educational efforts, the proportion of teenagers who use condoms at first intercourse could increase from 52% to 59%, while the proportion using no method might decrease from 41% to 33%.


Model for developing context-sensitive responses to vulnerability in research: managing ethical dilemmas faced by frontline research staff in Kenya.
Health research in low-resource settings often involves individuals and populations defined as 'vulnerable'. There is growing attention in the literature to the ethical dilemmas that frontline research staff face while conducting such research. However, there is little documented as to how research staff might support one another in identifying and handling these dilemmas in different contexts. Over the course of conducting empirical ethics research embedded in the Childhood Acute Illness & Nutrition Network, we developed an approach to examine and respond to the ethical issues and dilemmas faced by the study teams, particularly frontline staff. In this paper we describe the specific tools and approach we developed, which centred on regular cross-team ethics reflection sessions, and share lessons learnt. We suggest that all studies involving potentially vulnerable participants should incorporate activities and processes to support frontline staff in identifying, reflecting on and responding to ethical dilemmas, throughout studies. We outline the resources needed to do this and share piloted tools for further adaptation and evaluation. Such initiatives should complement and feed into-and certainly not in any way replace or substitute for-strong institutional ethics review, safeguarding and health and safety policies and processes, as well broader staff training and career support initiatives.


Osthole enhances glucose uptake through activation of AMP-activated protein kinase in skeletal muscle cells.
AMP-activated protein kinase (AMPK) is an energy sensor that regulates cellular metabolism. Activation of AMPK in skeletal muscles, the liver, and adipose tissues results in a favorable metabolic milieu for preventing and treating type 2 diabetes, i.e., decreased levels of circulating glucose, plasma lipids, and ectopic fat accumulation and enhanced insulin sensitivity. Osthole was extracted from a Chinese herbal medicine, and we found that it had glucose lowering activity in our previous study. However, the detailed glucose lowering mechanisms of osthole are still unclear. In this study, we used skeletal muscle cells to examine the underlying molecular mechanisms of osthole's glucose lowering activity. A Western blot analysis revealed that osthole significantly induced phosphorylation of AMPK and acetyl-CoA carboxylase (ACC). Next, we found that osthole significantly increased the level of translocation of glucose transporter 4 (GLUT4) to plasma membranes and glucose uptake in a dose-dependent manner. Osthole-induced glucose uptake was reversed by treatment with Compound C, an AMPK inhibitor, suggesting that osthole-induced glucose uptake was mediated in an AMPK-dependent manner. The increase in the AMP:ATP ratio was involved in osthole's activation of AMPK. Finally, we found that osthole counteracted hyperglycemia in mice with streptozotocin-induced diabetes. These results suggest that the increase in the AMP:ATP ratio by osthole triggered activation of the AMPK signaling pathway and led to increases in plasma membrane GLUT4 content and glucose uptake level. Therefore, osthole might have potential as an antidiabetic agent for treating diabetes.


Social acknowledgement as a predictor of post-traumatic stress and complicated grief after witnessing assisted suicide.
In Switzerland, right-to-die organizations such as Exit Deutsche Schweiz offer suicide assistance to their members. However, there is limited knowledge of the impact that witnessing assisted suicide has on the post-traumatic stress severity or the grief process of family members. Low perceived social acknowledgement may affect mental health. A cross-sectional survey of 85 family members who were present at an assisted suicide was conducted in December 2007. The Inventory of Complicated Grief and the Impact of Event Scale were used to assess symptoms of post-traumatic stress disorder (PTSD) and complicated grief (CG) at 14 to 24 months post-loss. Further, the Social Acknowledgement Questionnaire was used to examine the impact that the social environment's acknowledgement of the end-of-life decision had on respondents' mental health. As expected, social acknowledgement as a survivor was related to PTSD symptoms and CG. In particular, perceived general disapproval was strongly correlated with all outcome measures, whereas recognition was not significantly related to PTSD or CG (intrusion and avoidance). Family members of patients who use assisted suicide may hesitate to disclose the manner of death, and the community and societal environment may express strong views concerning the end-of-life decision. This can lead to increased levels of PTSD and CG.


Honing the Double-Edged Sword: Improving Human iPSC-Microglia Models.
Human induced Pluripotent Stem Cell (hiPSC) models are a valuable new tool for research into neurodegenerative diseases. Neuroinflammation is now recognized as a key process in neurodegenerative disease and aging, and microglia are central players in this. A plethora of hiPSC-derived microglial models have been published recently to explore neuroinflammation, ranging from monoculture through to xenotransplantation. However, combining physiological relevance, reproducibility, and scalability into one model is still a challenge. We examine key features of the in vitro microglial environment, especially media composition, extracellular matrix, and co-culture, to identify areas for improvement in current hiPSC-microglia models.