Establishment of a large semi-field system for experimental study of African malaria vector ecology and control in Tanzania.
Medical entomologists increasingly recognize that the ability to make inferences between laboratory experiments of vector biology and epidemiological trends observed in the field is hindered by a conceptual and methodological gap occurring between these approaches which prevents hypothesis-driven empirical research from being conducted on relatively large and environmentally realistic scales. The development of Semi-Field Systems (SFS) has been proposed as the best mechanism for bridging this gap. Semi-field systems are defined as enclosed environments, ideally situated within the natural ecosystem of a target disease vector and exposed to ambient environmental conditions, in which all features necessary for its life cycle completion are present. Although the value of SFS as a research tool for malaria vector biology is gaining recognition, only a few such facilities exist worldwide and are relatively small in size (< 100 m2). The establishment of a 625 m2 state-of-the-art SFS for large-scale experimentation on anopheline mosquito ecology and control within a rural area of southern Tanzania, where malaria transmission intensities are amongst the highest ever recorded, is described. A greenhouse frame with walls of mosquito netting and a polyethylene roof was mounted on a raised concrete platform at the Ifakara Health Institute. The interior of the SFS was divided into four separate work areas that have been set up for a variety of research activities including mass-rearing for African malaria vectors under natural conditions, high throughput evaluation of novel mosquito control and trapping techniques, short-term assays of host-seeking behaviour and olfaction, and longer-term experimental investigation of anopheline population dynamics and gene flow within a contained environment that simulates a local village domestic setting. The SFS at Ifakara was completed and ready for use in under two years. Preliminary observations indicate that realistic and repeatable observations of anopheline behaviour are obtainable within the SFS, and that habitat and climatic features representative of field conditions can be simulated within it. As work begins in the SFS in Ifakara and others around the world, the major opportunities and challenges to the successful application of this tool for malaria vector research and control are discussed.


The prostate: an increasing medical problem.
The prostate gland is the major site of medical problems in the U.S. male. Surgery for benign prostatic hyperplasia is the most common surgical procedure performed in males, and the total cost for this surgery exceeds 1 billion dollars per year. The incidence of and mortality from prostate cancer are increasing yearly. Prostate cancer now exceeds lung cancer as the most commonly diagnosed cancer in males and is the second leading cause of male cancer deaths. Of all tumors, the prevalence of prostate cancer increases the most rapidly with age. A shift in age distribution favoring an older population will lead to an increase in the number of patients diagnosed with prostate cancer in the United States. It is estimated that by the year 2000 there will be a 37% increase in prostate cancer deaths per year and a 90% increase in prostate cancer cases per year. Although the factors responsible for the development and progression of prostate cancer to a clinically manifest form are not known, there is evidence that environmental factors may play a role. It will be important for the future to focus on the etiological factors that may lead to a decrease in the prevalence of prostatic tumors.


Bullying victimization among 13 to 15-year-old school children: results from two comparative studies in 66 countries and regions.
to examine the prevalence of bullying victimization in 66 countries and territories from five continents based on data from two large international surveys: the 2001/2 Health Behavior in School-aged Children survey (HBSC) and the Global School-based Students Health Survey (GSHS). The surveys provide nationally representative, cross-sectional information on 13-15-year-old school children (N = 218,104). Bullying victimization, once or more within the past 2 months (HBSC)/30 days (GSHS). On average, 32.1% of the children were bullied at school at least once within the past 2 months in countries involved in the HBSC study and 37.4% of children were bullied at least one day within the past 30 days in countries involved in the GSHS study. In both surveys, a large variation in prevalence was found across countries. The lowest prevalence in the GSHS survey was observed in Tajikistan (7.1% for both boys and girls) and among HBSC countries in Sweden (16.0% for boys and 14.6% for girls) and the Czech Republic (17.1% for boys and 14.2% for girls). The highest prevalence for boys in the GSHS survey was found in Zimbabwe (Bulawayo) (70.2%) and for girls in Zambia (67.1%). In the HBSC survey, Lithuania had the highest prevalence of bullying victimization (62.7% for boys and 64.4% for girls). Victimization to bullying violates the fundamental rights of children; our study suggests that although many countries signed the United Nations Convention on the Rights of the Child, many have to work harder to fulfill the goals of the Convention.


ATP-sensitive K+ channel knockout compromises the metabolic benefit of exercise training, resulting in cardiac deficits.
Exercise training elicits a metabolic and cardiovascular response that underlies fitness. The molecular mechanisms that orchestrate this adaptive response and secure the wide-ranging gains of a regimented exercise program are poorly understood. Formed through association of the Kir6.2 pore and the sulfonylurea receptor, the stress-responsive ATP-sensitive K(+) channels (K(ATP) channels), with their metabolic-sensing capability and broad tissue expression, are potential candidates for integrating the systemic adaptive response to repetitive exercise. Here, the responses of mice lacking functional Kir6.2-containing K(ATP) channels (Kir6.2-KO) were compared with wild-type controls following a 28-day endurance swimming protocol. While chronic aquatic training resulted in lighter, leaner, and fitter wild-type animals, the Kir6.2-KO manifested less augmentation in exercise capacity and lacked metabolic improvement in body fat composition and glycemic handling with myocellular defects. Moreover, the repetitive stress of swimming unmasked a survival disadvantage in the Kir6.2-KO, associated with pathologic calcium-dependent structural damage in the heart and impaired cardiac performance. Thus, Kir6.2-containing K(ATP) channel activity is required for attainment of the physiologic benefits of exercise training without injury.


Risk factors for adverse life outcomes in fetal alcohol syndrome and fetal alcohol effects.
Clinical descriptions of patients with Fetal Alcohol Syndrome (FAS) and Fetal Alcohol Effects (FAE) suggest major problems with adaptive behavior. Five operationally defined adverse outcomes and 18 associated risk/protective factors were examined using a Life History Interview with knowledgeable informants of 415 patients with FAS or FAE (median age 14 years, range 6-51; median IQ 86, range 29-126). Eighty percent of these patients were not raised by their biological mothers. For adolescents and adults, the life span prevalence was 61% for Disrupted School Experiences, 60% for Trouble with the Law, 50% for Confinement (in detention, jail, prison, or a psychiatric or alcohol/drug inpatient setting), 49% for Inappropriate Sexual Behaviors on repeated occasions, and 35% for Alcohol/Drug Problems. The odds of escaping these adverse life outcomes are increased 2- to 4-fold by receiving the diagnosis of FAS or FAE at an earlier age and by being reared in good stable environments.


Host orientation using volatiles in the phoretic nematode Caenorhabditis japonica.
Host orientation is the most important step in host-searching nematodes; however, information on direct cues from hosts to evoke this behaviour is limited. Caenorhabditis japonica establishes a species-specific phoresy with Parastrachia japonensis. Dauer larvae (DL), the non-feeding and phoretic stage of C. japonica, are predominantly found on female phoretic hosts, but the mechanisms underlying the establishment of this phoresy remain unknown. To determine whether C. japonica DL are able to recognize and orient themselves to a host using a volatile cue from the host, we developed a Y-tube olfactory assay system in which C. japonica DL were significantly attracted to the air from P. japonensis but not to the air from three other insects or to CO2. These results demonstrated that C. japonica DL utilize volatiles for host recognition and orientation and that the presence of a specific volatile kairomone released by the host attracts C. japonica DL.


Randomised trial of endoscopic versus percutaneous stent insertion in malignant obstructive jaundice.
Patients with biliary obstruction due to malignant disease, and judged unfit for open operation, were randomised to have a biliary stent inserted either endoscopically via the papilla of Vater or percutaneously. Analysis after 75 patients had been entered showed that the endoscopic method had a significantly higher success rate for relief of jaundice (81% versus 61%, p = 0.017) and a significantly lower 30-day mortality (15% versus 33%, p = 0.016). The higher mortality after percutaneous stents was due to complications associated with liver puncture (haemorrhage and bile leaks). When stenting is indicated in elderly and frail patients the endoscopic method should be tried first.


Environmental Risk Assessment Strategy for Nanomaterials.
An Environmental Risk Assessment (ERA) for nanomaterials (NMs) is outlined in this paper. Contrary to other recent papers on the subject, the main data requirements, models and advancement within each of the four risk assessment domains are described, i.e., in the: (i) materials, (ii) release, fate and exposure, (iii) hazard and (iv) risk characterisation domains. The material, which is obviously the foundation for any risk assessment, should be described according to the legislatively required characterisation data. Characterisation data will also be used at various levels within the ERA, e.g., exposure modelling. The release, fate and exposure data and models cover the input for environmental distribution models in order to identify the potential (PES) and relevant exposure scenarios (RES) and, subsequently, the possible release routes, both with regard to which compartment(s) NMs are distributed in line with the factors determining the fate within environmental compartment. The initial outcome in the risk characterisation will be a generic Predicted Environmental Concentration (PEC), but a refined PEC can be obtained by applying specific exposure models for relevant media. The hazard information covers a variety of representative, relevant and reliable organisms and/or functions, relevant for the RES and enabling a hazard characterisation. The initial outcome will be hazard characterisation in test systems allowing estimating a Predicted No-Effect concentration (PNEC), either based on uncertainty factors or on a NM adapted version of the Species Sensitivity Distributions approach. The risk characterisation will either be based on a deterministic risk ratio approach (i.e., PEC/PNEC) or an overlay of probability distributions, i.e., exposure and hazard distributions, using the nano relevant models.


Strength and Diversity of Inhibitory Signaling Differentiates Primate Anterior Cingulate from Lateral Prefrontal Cortex.
The lateral prefrontal cortex (LPFC) and anterior cingulate cortex (ACC) of the primate play distinctive roles in the mediation of complex cognitive tasks. Compared with the LPFC, integration of information by the ACC can span longer timescales and requires stronger engagement of inhibitory processes. Here, we reveal the synaptic mechanism likely to underlie these differences using in vitro patch-clamp recordings of synaptic events and multiscale imaging of synaptic markers in rhesus monkeys. Although excitatory synaptic signaling does not differ, the level of synaptic inhibition is much higher in ACC than LPFC layer 3 pyramidal neurons, with a significantly higher frequency (∼6×) and longer duration of inhibitory synaptic currents. The number of inhibitory synapses and the ratio of cholecystokinin to parvalbumin-positive inhibitory inputs are also significantly higher in ACC compared with LPFC neurons. Therefore, inhibition is functionally and structurally more robust and diverse in ACC than in LPFC, resulting in a lower excitatory: inhibitory ratio and a greater dynamic range for signal integration and network oscillation by the ACC. These differences in inhibitory circuitry likely underlie the distinctive network dynamics in ACC and LPC during normal and pathological brain states.SIGNIFICANCE STATEMENT The lateral prefrontal cortex (LPFC) and anterior cingulate cortex (ACC) play temporally distinct roles during the execution of cognitive tasks (rapid working memory during ongoing tasks and long-term memory to guide future action, respectively). Compared with LPFC-mediated tasks, ACC-mediated tasks can span longer timescales and require stronger engagement of inhibition. This study shows that inhibitory signaling is much more robust and diverse in the ACC than in the LPFC. Therefore, there is a lower excitatory: inhibitory synaptic ratio and a greater dynamic range for signal integration and oscillatory behavior in the ACC. These significant differences in inhibitory synaptic transmission form an important basis for the differential timing of cognitive processing by the LPFC and ACC in normal and pathological brain states.


Urea-induced unfolding of the alpha subunit of tryptophan synthase: evidence for a multistate process.
The urea-induced unfolding of the alpha subunit of tryptophan synthase from E. coli was monitored by optical spectroscopy and by urea-gradient gel electrophoresis. Three independent lines of evidence support the conclusion that one or more stable intermediates are present in this process: (i) Satisfactory fits of the equilibrium unfolding transitions obtained from difference spectroscopy at 286 nm and circular dichroism spectroscopy at 222 nm require a model which involves a stable intermediate in addition to the native and unfolded forms. (ii) Kinetic studies of the change in the extinction coefficient at 286 nm show that while the unfolding is well described by a single exponential change the refolding kinetics are complex. The nature of the dependence of the refolding kinetics on the initial concentration of urea supports the conclusion that at least one stable intermediate exists. (iii) The patterns obtained from urea-gradient gel electrophoresis experiments on the alpha subunit show that at least one and possibly two stable intermediates are involved; the intermediates have markedly different degrees of compactness. A kinetic model for the folding of the alpha subunit, consistent with all of these results, can be formulated.


Accuracy and Reliability of a Handheld, Nonmydriatic Fundus Camera for the Remote Detection of Optic Disc Edema.
Optic disc edema can be an important indicator of serious neurological disease, but is poorly detected using the direct ophthalmoscope. Portable fundus photography may overcome this difficulty. The purpose of this study was to determine the sensitivity and specificity of a handheld, nonmydriatic fundus camera for the detection of optic disc edema. Retrospective review of nonmydriatic optic disc photographs taken with a portable fundus camera (Pictor Plus; Volk Optical, Mentor, OH) from the University of Michigan Neuro-Ophthalmology Clinics. We included 103 consecutive eyes with optic disc edema and 103 consecutive eyes without optic disc edema of 109 patients. Four masked neuro-ophthalmologists graded a single photograph of each optic disc presented in randomized order and documented the presence of optic disc edema. Sensitivity and specificity of graders' photographic interpretation was compared with clinical examinations. Reliability of assessments within and between graders was determined using kappa statistics. The sensitivity and specificity for detection of optic disc edema were 71.8-92.2% and 81.6-95.2%, respectively. Photos were found to be ungradable in 0-8.3% of cases. The intergrader reliabilities ranged from 0.60 [95% confidence interval (CI): 0.52-0.67] to 0.72 (95% CI: 0.66-0.77). Intragrader reliability ranged from 0.76 (95% CI: 0.63-0.92) to 0.82 (95% CI: 0.69-0.95). Photographs taken with portable, nonmydriatic technology met threshold sensitivity and specificity for remote screening for optic disc edema when performed by most, but not all graders. Reliability between graders was moderate-strong and strong within individual providers. Portable photography holds promise for use in remote screening of optic disc edema.


The effect of gibberellic acid alone and when combined with [2-chloroethyl]-trimethyl ammonium chloride on the growth and alkaloid content of Solanum laciniatum aiton.
Solanum laciniatum Aiton treated with Gibberellic acid (GA3), [2-chloroethyl]-trimethyl ammonium chloride (CCC), and their combinations at early and late stages of growth showed that early application of 2000 ppm CCC produced the greatest stem, leaves, and whole plant dry weight, followed by the combination of early application of 1000 ppm CCC and late application of 100 ppm GA3; whereas all GA3 treatments decreased the dry weight production compared with the controls. Regarding the glyco-alkaloids, the highest percentage was obtained from the whole plant by early applications of both strengths of GA3 compared with other treatments or the controls. On the other hand, early application of 1000 ppm CCC and also late application of 50 ppm GA3 when combined with early application of either 1000 ppm or 2000 ppm CCC produced greater alkaloid percentage yields in stem, leaves, and whole plants more than other treatments or the controls. However, early application of 2000 ppm CCC produced the highest content of alkaloids in leaves and whole plants; this was followed by yields from early application of 2000 ppm CCC and 50 ppm GA3; yields from early application of 100 ppm GA3 and 1000 ppm CCC; and yields from early application of 1000 ppm CCC +50 ppm GA3.


Meta-analysis for genome-wide association study identifies multiple variants at the BIN1 locus associated with late-onset Alzheimer's disease.
Recent GWAS studies focused on uncovering novel genetic loci related to AD have revealed associations with variants near CLU, CR1, PICALM and BIN1. In this study, we conducted a genome-wide association study in an independent set of 1034 cases and 1186 controls using the Illumina genotyping platforms. By coupling our data with available GWAS datasets from the ADNI and GenADA, we replicated the original associations in both PICALM (rs3851179) and CR1 (rs3818361). The PICALM variant seems to be non-significant after we adjusted for APOE e4 status. We further tested our top markers in 751 independent cases and 751 matched controls. Besides the markers close to the APOE locus, a marker (rs12989701) upstream of BIN1 locus was replicated and the combined analysis reached genome-wide significance level (p = 5E-08). We combined our data with the published Harold et al. study and meta-analysis with all available 6521 cases and 10360 controls at the BIN1 locus revealed two significant variants (rs12989701, p = 1.32E-10 and rs744373, p = 3.16E-10) in limited linkage disequilibrium (r² = 0.05) with each other. The independent contribution of both SNPs was supported by haplotype conditional analysis. We also conducted multivariate analysis in canonical pathways and identified a consistent signal in the downstream pathways targeted by Gleevec (P = 0.004 in Pfizer; P = 0.028 in ADNI and P = 0.04 in GenADA). We further tested variants in CLU, PICALM, BIN1 and CR1 for association with disease progression in 597 AD patients where longitudinal cognitive measures are sufficient. Both the PICALM and CLU variants showed nominal significant association with cognitive decline as measured by change in Clinical Dementia Rating-sum of boxes (CDR-SB) score from the baseline but did not pass multiple-test correction. Future experiments will help us better understand potential roles of these genetic loci in AD pathology.


Treatment of a chronically infected nasal silicone prosthesis with continuous antibiotic irrigation and gentamicin-impregnated polymethylmethacrylate beads.
Infected nasal alloplasts in revision rhinoplasty can be a complex problem, as timing between implant removal and reconstruction is the major limiting factor. Delaying reconstruction can result in loss of mechanical support, a constricted nose, and in severe cases, complete nasal airway collapse and respiratory compromise. In this case report, we describe a novel surgical approach for the management of a chronically infected nasal implant combining techniques used to treat biomaterial-associated infections: antibiotic-impregnated polymethymethacrylate beads and a continuous catheter-based antibiotic irrigation system. We report a case of a chronic alloplastic-associated infection following nasal reconstruction using a silicone implant. We utilized a two-staged approach. The involved nasal implant was removed and replaced temporarily with gentamicin-impregnated polymethymethacrylate beads and a continuous closed irrigation and drainage system with local and parenteral delivery of antibiotics. Both modalities allowed for complete eradication of the infection. In addition, the gentamicin beads provided sufficient mechanical support in order to minimize the risk of skin contracture. Twelve days after her initial surgery, nasal reconstruction was performed using a cadaver bone graft. The patient was followed for two years postoperatively and has shown good results with no evidence of skin contracture or recurrent infection. This technique may allow for shorter delay in revision surgery and reduce the risk of long-term complications without compromising functional and aesthetic outcomes.


Comparison of quartz and sapphire optical chambers for infrared laser sealing of vascular tissues using a reciprocating, side-firing optical fiber: Simulations and experiments.
Infrared (IR) lasers are being tested as an alternative to radiofrequency (RF) and ultrasonic (US) surgical devices for hemostatic sealing of vascular tissues. In previous studies, a side-firing optical fiber with elliptical IR beam output was reciprocated, producing a linear IR laser beam pattern for uniform sealing of blood vessels. Technical challenges include limited field-of-view of vessel position within the metallic device jaws, and matching fiber scan length to variable vessel sizes. A transparent jaw may improve visibility and enable custom treatment. Quartz and sapphire square optical chambers (2.7 × 2.7 × 25 [mm3 ] outer dimensions) were tested, capable of fitting into a 5-mm-OD laparoscopic device. A 1470 nm laser was used for optical transmission studies. Razor blade scans and an IR beam profiler acquired fiber (550-µm-core/0.22NA) output beam profiles. Thermocouples recorded peak temperatures and cooling times on internal and external chamber surfaces. Optical fibers with angle polished distal tips delivered 94% of light at a 90° angle. Porcine renal arteries with diameters of 3.4 ± 0.7 mm (n = 13) for quartz and 3.2 ± 0.7 mm (n = 14) for sapphire chambers (p > 0.05), were sealed using 30 W for 5 s. Reflection losses at material/air interfaces were 3.3% and 7.4% for quartz and sapphire. Peak temperatures on the external chamber surface averaged 74 ± 8°C and 73 ± 10°C (p > 0.05). Times to cool down to 37°C measured 13 ± 4 s and 27 ± 7 s (p < 0.05). Vessel burst pressures (BP) averaged 883 ± 393 mmHg and 412 ± 330 mmHg (p < 0.05). For quartz, 13/13 (100%) vessels were sealed (BP > 360 mmHg), versus 9/14 (64%) for sapphire. Computer simulations for the quartz chamber yielded peak temperatures (78°C) and cooling times (16 s) similar to experiments. Quartz is an inexpensive material for use in a laparoscopic device jaw, providing more consistent vessel seals and faster cooling times than sapphire and current RF and US devices.


[Effect of neurotropin on chronic headaches in children].
We report the cases of 2 girls, aged 13 and 15 years, who experienced exacerbation of migraine at 2-3 years after its onset. The chronic headaches were refractory to various anti-migraine and analgesic agents, persisted for 3-6 months in each patient, and rendered them unable to attend school. These headaches commonly manifested with symptoms of orthostatic dysregulation and paroxysmal back/limb pain. In addition, T2-weighted magnetic resonance images showed dots of highly intense signals in the bilateral cerebral white matter. Neurotropin, a non-protein extract isolated from the dermis of rabbits and inoculated with the vaccinia virus, induced prompt significant effects on the headache and back/limbs pain in both the girls. The symptom of orthostatic dysregulation was also gradually ameliorated after the resolution of the headache. Neurotropin has an analgesic action that is not common to other drugs since it enhances the descending pain inhibitory systems, and its effect has been reported in cases of post-herpetic and other neuralgia as well as in cases of primary headaches in adulthood. This agent may also be promising for the treatment of intractable primary headaches during childhood.


Better Retention of Abatacept Is Associated with High Rheumatoid Factor: A Five-Year Follow-Up Study of Patients with Rheumatoid Arthritis.
Recently, biological disease-modifying antirheumatic drugs (bDMARDs) have revolutionized the treatment of rheumatoid arthritis (RA) and provided patients with a higher chance of achieving clinical remission. Among them, abatacept (ABT), which selectively inhibits T cell activation through blocking costimulation signal, has been reported efficacious in controlling disease activity. Previous studies have shown that ABT has a high retention rate of up to three years with tolerable adverse events; however, it remains unclear whether this is maintained in the longer term. Here we conducted a retrospective five-year follow-up study to explore prognostic factors concerning better retention. In total, 98 patients who were treated with ABT from May 2011 to July 2019 in Osaki Citizen Hospital were enrolled, including 73 female patients (74.5%). The Kaplan-Meier method was used to estimate the retention rate of ABT. The mean age of ABT initiation was 72.1 years. Concomitant methotrexate was prescribed for 39 patients, and ABT was used as the first-line bDMARD for 65 patients. Rheumatoid factor (RF) was positive in 79 patients. One-, three-, and five-year retention rates of ABT were 83.3%, 66.2%, and 62.7%, respectively. Approximately two-thirds of discontinuation resulted from an inadequate response. Multivariate logistic regression analysis revealed that positive RF was associated with better drug retention. Receiver operating characteristics analysis showed that patients with high RF (≥ 45 IU/mL) had better retention rate of ABT. In conclusion, ABT shows high retention rate among patients with positive RF. The present study may provide better insights when selecting bDMARDs.


State Olmstead litigation and the Affordable Care Act.
Over the past two decades, major efforts have been undertaken to expand access to Medicaid home and community-based services (HCBS) for the elderly and disabled. Despite this, many states still have long waiting lists for HCBS. Using data collected, this study examined the trends in Olmstead and related cases against states between 1999 and 2011. The findings show there were 131 cases filed during the period, and 90 cases were resolved through court rulings and settlements. These court cases have played an important role in encouraging states to expand access to HCBS programs and to transfer individuals out of institutions.


The relationship between familial resemblance and sexual attraction: an update on Westermarck, Freud, and the incest taboo.
Foundational principles of evolutionary theory predict that inbreeding avoidance mechanisms should exist in all species--including humans--in which close genetic relatives interact during periods of sexual maturity. Voluminous empirical evidence, derived from diverse taxa, supports this prediction. Despite such results, Fraley and Marks claim to provide evidence that humans are sexually attracted to close genetic relatives and that such attraction is held in check by cultural taboos. Here, the authors show that Fraley and Marks, in their search for an alternate explanation of inbreeding avoidance, misapply theoretical constructs from evolutionary biology and social psychology, leading to an incorrect interpretation of their results. The authors propose that Fraley and Marks's central findings can be explained in ways consistent with existing evolutionary models of inbreeding avoidance. The authors conclude that appropriate application of relevant theory and stringent experimental design can generate fruitful investigations into sexual attraction, inbreeding avoidance, and incest taboos.


Beneficial effects of yohimbine on posthypoxic recovery of cardiac function and myocardial metabolism in isolated perfused rabbit hearts.
The present study was undertaken to elucidate the possible actions of yohimbine on cardiac function and metabolism in the hypoxic and subsequently reoxygenated myocardium. For this purpose, rabbit hearts were perfused for 20 min under hypoxic conditions, followed by 45 min reoxygenated perfusion, and their functional and metabolic alterations with and without yohimbine treatment were examined. Hypoxia induced cessation of cardiac contractile force, rise in resting tension and depletion of tissue high-energy phosphates, which were poorly recovered by subsequent reoxygenation. Hypoxia also induced release of creatine kinase and ATP metabolites from perfused hearts and increases in tissue calcium and sodium contents, which were further enhanced upon subsequent reoxygenation. When hypoxic hearts were treated with 3 to 30 microM yohimbine, several beneficial effects were observed in a concentration-dependent manner. This included enhancement of posthypoxic recovery of contractile function and suppression of the hypoxia- and reoxygenation-induced rise in resting tension. Hypoxia/reoxygenation-induced release of ATP metabolites was inhibited and restoration of myocardial high-energy phosphates enhanced. Inhibition of reoxygenation-induced rise in tissue calcium and sodium and creatine kinase release were also noted. The findings suggest that suppression of transmembrane flux of ions, substrates and enzymes during hypoxia/reoxygenation plays a role in the posthypoxic functional and metabolic recovery. Yohimbine (3-30 microM) significantly depressed the maximal stimulus frequency the left atria could follow. These results suggest a close relationship between depression in the maximal driving frequency of atria and enhancement of the posthypoxic contractile and metabolic recovery of perfused hearts.(ABSTRACT TRUNCATED AT 250 WORDS)


Adverse childhood experiences of low-income urban youth.
Current assessments of adverse childhood experiences (ACEs) may not adequately encompass the breadth of adversity to which low-income urban children are exposed. The purpose of this study was to identify and characterize the range of adverse childhood experiences faced by young adults who grew up in a low-income urban area. Focus groups were conducted with young adults who grew up in low-income Philadelphia neighborhoods. Using the nominal group technique, participants generated a list of adverse childhood experiences and then identified the 5 most stressful experiences on the group list. The most stressful experiences identified by participants were grouped into a ranked list of domains and subdomains. Participants identified a range of experiences, grouped into 10 domains: family relationships, community stressors, personal victimization, economic hardship, peer relationships, discrimination, school, health, child welfare/juvenile justice, and media/technology. Included in these domains were many but not all of the experiences from the initial ACEs studies; parental divorce/separation and mental illness were absent. Additional experiences not included in the initial ACEs but endorsed by our participants included single-parent homes; exposure to violence, adult themes, and criminal behavior; personal victimization; bullying; economic hardship; and discrimination. Gathering youth perspectives on childhood adversity broadens our understanding of the experience of stress and trauma in childhood. Future work is needed to determine the significance of this broader set of adverse experiences in predisposing children to poor health outcomes as adults.


Neurocysticercosis: relationship between the developmental stage of metacestode present and the titre of specific IgG in the cerebrospinal fluid.
In double-blind, immunological assays, samples of cerebrospinal fluid (CSF) from 141 patients with neurocysticercosis (NCC) or other neurological disorders were tested for IgG reacting with the excretory/secretory (E/S) antigens of Taenia solium metacestodes. The results for the cases of NCC were then correlated with the developmental stage of the metacestodes present in each case, as assessed by computerized tomography and magnetic-resonance imaging. In the ELISA first used, the samples of CSF from most (88%) of the patients with the vesicular stage of NCC (some of whom also had the degenerate and/or calcified metacestodes) were found to contain the specific IgG. In electro-immunotransfer blot (EITB) assays, three of the E/S antigens, of 95, 49 and 29 kDA, were recognized by 86%-100% of the ELISA-positive CSF. When these three antigens were isolated and tested, as a pool, against all the CSF samples in double-blind ELISA, almost all (96.6%) of the CSF samples from patients with metacestodes at the vesicular stage were recognized. In the detection of individuals with vesicular metacestodes, the assay based on the three isolated antigens was significantly more sensitive than that based on the crude extract of E/S antigens (P < 0.05). In EITB assays based on the three antigens, the isolated proteins were again recognized by IgG in the CSF samples from those with vesicular metacestodes, but without the background 'noise' seen with the crude extract. In every assay employed, none of the CSF samples from NCC cases who only harboured degenerative and/or calcified metacestodes and none of those from patients who had other neurological disorders gave a positive result. The use in ELISA and EITB of antigens purified from crude extracts of metacestode E/S proteins could improve the immunodiagnosis of the vesicular stage of NCC, and allow better evaluation of NCC cases both pre- and post-treatment.


Deletion of the muscle enriched lncRNA Oip5os1 induces atrial dysfunction in male mice with diabetes.
Long ncRNAs (lncRNAs) have been shown to play a biological and physiological role in various tissues including the heart. We and others have previously established that the lncRNA Oip5os1 (1700020I14Rik, OIP5-AS1, Cyrano) is enriched in striated muscles, and its deletion in mice leads to defects in both skeletal and cardiac muscle function. In the present study, we investigated the impact of global Oip5os1 deletion on cardiac function in the setting of streptozotocin (STZ)-induced diabetes. Specifically, we studied male WT and KO mice with or without diabetes for 24 weeks, and phenotyped animals for metabolic and cardiac endpoints. Independent of genotype, diabetes was associated with left ventricular diastolic dysfunction based on a fall in E'/A' ratio. Deletion of Oip5os1 in a setting of diabetes had no significant impact on ventricular function or ventricular weight, but was associated with left atrial dysfunction (reduced fractional shortening) and myopathy which was associated with anesthesia intolerance and premature death in the majority of KO mice tested during cardiac functional assessment. This atrial phenotype was not observed in WT diabetic mice. The most striking molecular difference was a reduction in the metabolic regulator ERRalpha in the atria of KO mice compared with WT mice. There was also a trend for a reduction in Serca2a. These findings highlight Oip5os1 as a gene of interest in aspects of atrial function in the setting of diabetes, highlighting an additional functional role for this lncRNA in cardiac pathological settings.


Stable gene selection from microarray data via sample weighting.
Feature selection from gene expression microarray data is a widely used technique for selecting candidate genes in various cancer studies. Besides predictive ability of the selected genes, an important aspect in evaluating a selection method is the stability of the selected genes. Experts instinctively have high confidence in the result of a selection method that selects similar sets of genes under some variations to the samples. However, a common problem of existing feature selection methods for gene expression data is that the selected genes by the same method often vary significantly with sample variations. In this work, we propose a general framework of sample weighting to improve the stability of feature selection methods under sample variations. The framework first weights each sample in a given training set according to its influence to the estimation of feature relevance, and then provides the weighted training set to a feature selection method. We also develop an efficient margin-based sample weighting algorithm under this framework. Experiments on a set of microarray data sets show that the proposed algorithm significantly improves the stability of representative feature selection algorithms such as SVM-RFE and ReliefF, without sacrificing their classification performance. Moreover, the proposed algorithm also leads to more stable gene signatures than the state-of-the-art ensemble method, particularly for small signature sizes.


Synergistic Action of a Metalloprotease and a Serine Protease from Fusarium oxysporum f. sp. lycopersici Cleaves Chitin-Binding Tomato Chitinases, Reduces Their Antifungal Activity, and Enhances Fungal Virulence.
As part of their defense strategy against fungal pathogens, plants secrete chitinases that degrade chitin, the major structural component of fungal cell walls. Some fungi are not sensitive to plant chitinases because they secrete chitin-binding effector proteins that protect their cell wall against these enzymes. However, it is not known how fungal pathogens that lack chitin-binding effectors overcome this plant defense barrier. Here, we investigated the ability of fungal tomato pathogens to cleave chitin-binding domain (CBD)-containing chitinases and its effect on fungal virulence. Four tomato CBD chitinases were produced in Pichia pastoris and were incubated with secreted proteins isolated from seven fungal tomato pathogens. Of these, Fusarium oxysporum f. sp. lycopersici, Verticillium dahliae, and Botrytis cinerea were able to cleave the extracellular tomato chitinases SlChi1 and SlChi13. Cleavage by F. oxysporum removed the CBD from the N-terminus, shown by mass spectrometry, and significantly reduced the chitinase and antifungal activity of both chitinases. Both secreted metalloprotease FoMep1 and serine protease FoSep1 were responsible for this cleavage. Double deletion mutants of FoMep1 and FoSep1 of F. oxysporum lacked chitinase cleavage activity on SlChi1 and SlChi13 and showed reduced virulence on tomato. These results demonstrate the importance of plant chitinase cleavage in fungal virulence.


Nonradioactive iodide effectively induces apoptosis in genetically modified lung cancer cells.
We assessed a nonradioactive approach to induce apoptosis in non-small cell lung cancer by a novel iodide uptake and retention mechanism. To enhance tumor apoptosis, we transduced non-small cell lung cancer cells with retroviral vectors containing the sodium iodide symporter (NIS) and thyroperoxidase (TPO) genes. Expression of NIS and TPO facilitated concentration of iodide in tumors. As a consequence of the marked increase in intracellular levels of iodide, apoptosis was seen in >95% of NIS/TPO-modified lung cancer cells. Intraperitoneal injection of potassium iodide resulted in significant tumor volume reduction in NIS/TPO-modified tumor xenografts without apparent adverse effects in SCID mice. Iodide induced an increase in the level of reactive oxygen species. Iodide-induced apoptosis is sensitive to N-acetylcysteine inhibition, suggesting an important role by reactive oxygen species in this apoptotic process. In addition, iodide-induced apoptosis is associated with overexpression of CDKN1A (p21/Waf1)and down-regulation of survivin at both mRNA and protein levels. This is the first report demonstrating that a therapeutic dose of nonradioactive iodide has potent efficacy and high selectivity against lung cancer when used in combination with genetic modification of cancer cells to express the NIS/TPO genes.


Floquet analysis for vibronically modulated electron tunneling.
Electron tunneling provides the primary reaction channel for electron transfer (ET) in many molecular systems. The analysis of such systems therefore requires the consideration of electronic coherence and interference effects. A model system for which tunneling may be either symmetry forbidden or allowed is considered here in the presence of a driving infrared (IR) field. It was previously shown that inelastic tunneling allows ET in the symmetry forbidden system via vibronic interactions. We show here that explicit considerations of IR interactions with these systems further changes the ET kinetics. Analysis in the framework of Floquet theory reveals that interaction with an IR field may increase the probability of inelastic tunneling and thus enhance the ET rate for a system in which elastic ET is forbidden. It is shown that IR driving of a nuclear oscillator promotes the oscillator into excited states that couple more strongly to the tunneling electron. Furthermore, it is shown that IR driving may suppress the ET rate in this same system, depending on system energetics. In a model where elastic tunneling is symmetry allowed, we examine vibronic modulation of ET in the Floquet framework. ET rates are computed for symmetry allowed and forbidden model systems, and vibronic interactions are found to suppress or enhance ET in both systems. The inelastic ET rate may be enhanced over 4 orders of magnitude in the symmetry disfavored case or suppressed by 15% in the same system. Effects of IR on ET rates in the symmetry-allowed system are weaker with enhancements up to 34% over the undriven rate and suppression of about 3% with IR driving present. This study is the first theoretical and computational exploration of ET rate control by IR irradiation of the bridge.


Sequencing and genetic variation of multidrug resistance plasmids in Klebsiella pneumoniae.
The development of multidrug resistance is a major problem in the treatment of pathogenic microorganisms by distinct antimicrobial agents. Characterizing the genetic variation among plasmids from different bacterial species or strains is a key step towards understanding the mechanism of virulence and their evolution. We applied a deep sequencing approach to 206 clinical strains of Klebsiella pneumoniae collected from 2002 to 2008 to understand the genetic variation of multidrug resistance plasmids, and to reveal the dynamic change of drug resistance over time. First, we sequenced three plasmids (70 Kb, 94 Kb, and 147 Kb) from a clonal strain of K. pneumoniae using Sanger sequencing. Using the Illumina sequencing technology, we obtained more than 17 million of short reads from two pooled plasmid samples. We mapped these short reads to the three reference plasmid sequences, and identified a large number of single nucleotide polymorphisms (SNPs) in these pooled plasmids. Many of these SNPs are present in drug-resistance genes. We also found that a significant fraction of short reads could not be mapped to the reference sequences, indicating a high degree of genetic variation among the collection of K. pneumoniae isolates. Moreover, we identified that plasmid conjugative transfer genes and antibiotic resistance genes are more likely to suffer from positive selection, as indicated by the elevated rates of nonsynonymous substitution. These data represent the first large-scale study of genetic variation in multidrug resistance plasmids and provide insight into the mechanisms of plasmid diversification and the genetic basis of antibiotic resistance.


High proportion of fluoroquinolone-resistant Mycobacterium tuberculosis isolates with novel gyrase polymorphisms and a gyrA region associated with fluoroquinolone susceptibility.
Fluoroquinolone resistance in Mycobacterium tuberculosis can be conferred by mutations in gyrA or gyrB. The prevalence of resistance mutations outside the quinolone resistance-determining region (QRDR) of gyrA or gyrB is unclear, since such regions are rarely sequenced. M. tuberculosis isolates from 1,111 patients with newly diagnosed culture-confirmed tuberculosis diagnosed in Tennessee from 2002 to 2009 were screened for phenotypic ofloxacin resistance (>2 μg/ml). For each resistant isolate, two ofloxacin-susceptible isolates were selected: one with antecedent fluoroquinolone exposure and one without. The complete gyrA and gyrB genes were sequenced and compared with M. tuberculosis H37Rv. Of 25 ofloxacin-resistant isolates, 11 (44%) did not have previously reported resistance mutations. Of these, 10 had novel polymorphisms: 3 in the QRDR of gyrA, 1 in the QRDR of gyrB, and 6 outside the QRDR of gyrA or gyrB; 1 did not have any gyrase polymorphisms. Polymorphisms in gyrA codons 1 to 73 were more common in fluoroquinolone-susceptible than in fluoroquinolone-resistant strains (20% versus 0%; P = 0.016). In summary, almost half of fluoroquinolone-resistant M. tuberculosis isolates did not have previously described resistance mutations, which has implications for genotypic diagnostic tests.


Correlation of parenting style and pediatric behavior guidance strategies in the dental setting: preliminary findings.
The aim of this study was to investigate the influence of parenting style on the choice of proper behavior guidance strategies in pedodontics. Seventy-two children aged between 4 and 6 years (mean 5.12 years) with carious primary mandibular molars were selected. The Primary Caregivers' Practices Report (PCPR) was used to quantify authoritarian, permissive and authoritative aspects of the caregivers' parenting style. After inferior alveolar nerve block, carious lesions were removed and the teeth were restored using amalgam. The children's behavior during operation was assessed according to the sound, eye, and motor (SEM) scale. Communicative guidance, advance behavior guidance, parental separation, and deferred treatment were used for behavior management. The dominant authoritative score was observed in 50% of parents, permissive in 37.5%, and authoritarian in 12.5%. The mean SEM score in children belonging to authoritative parents was significantly lower than in children of permissive and of authoritarian parents (p<0.05). Communicative guidance was necessary for all patients regardless of parenting style. Advanced behavior guidance (protective stabilization) was applied in 16.7% of cases in the authoritative category and in 100% in the permissive and authoritarian categories. The use of restrictive devices (7.4%) and sedation (3.7%) was limited to the permissive category. Parental separation (40.7%) and deferred treatment (3.7%) were performed only in the permissive category. This study provides preliminary evidence that a child's reaction to restorative dental procedures is influenced by the nature of the caregiver's parenting style.


Somatic influences on subjective well-being and affective disorders: the convergence of thermosensory and central serotonergic systems.
Current theories suggest that the brain is the sole source of mental illness. However, affective disorders, and major depressive disorder (MDD) in particular, may be better conceptualized as brain-body disorders that involve peripheral systems as well. This perspective emphasizes the embodied, multifaceted physiology of well-being, and suggests that afferent signals from the body may contribute to cognitive and emotional states. In this review, we focus on evidence from preclinical and clinical studies suggesting that afferent thermosensory signals contribute to well-being and depression. Although thermoregulatory systems have traditionally been conceptualized as serving primarily homeostatic functions, increasing evidence suggests neural pathways responsible for regulating body temperature may be linked more closely with emotional states than previously recognized, an affective warmth hypothesis. Human studies indicate that increasing physical warmth activates brain circuits associated with cognitive and affective functions, promotes interpersonal warmth and prosocial behavior, and has antidepressant effects. Consistent with these effects, preclinical studies in rodents demonstrate that physical warmth activates brain serotonergic neurons implicated in antidepressant-like effects. Together, these studies suggest that (1) thermosensory pathways interact with brain systems that control affective function, (2) these pathways are dysregulated in affective disorders, and (3) activating warm thermosensory pathways promotes a sense of well-being and has therapeutic potential in the treatment of affective disorders.


[Medication administration errors at an emergency service: knowing to decrease risk].
TThere is a great potential risk of errors in the emergency services and the studies carried out present heterogeneous results and methodology. The aim was to calculate the prevalence and characteristics, types and consequences for the patient, of medications administration errors in patients who attended the emergency department (ED) of a tertiary hospital, and to propose improvement strategies to prevent them. A cross-sectional study of direct observation in the emergency areas of internal medicine and observation of adult patients was conducted in September 2016. Data collection was conducted by 4, nursing professional's observers from the ED. Medication errors reported by professionals to the hospital's voluntary reporting system were also analysed. The number of medication administration errors observed was 150 (19%) versus the number of medication administration errors reported to the hospital's notification system, which was 14, from 2013 to 2016. The most frequent type of error was incorrect administration rate (67%), followed by preparation errors (16%) and incorrect time administration (14%). The medications that accumulated the most errors were furosemide and methylprednisolone. None of the errors involved damage to the patient. Descriptive statistics was used to summarize the results of the study. The prevalence of medication administration errors in the ED is high. Knowing about them is effective to undertake specific improvement actions that may influence prevention, increasing patient safety.


IL-18-induced HIF-1α in ILC3s ameliorates the inflammation of C. rodentium-induced colitis.
Group 3 innate lymphoid cells (ILC3s) are vital for defending tissue barriers from invading pathogens. Hypoxia influences the production of intestinal ILC3-derived cytokines by activating HIF. Yet, the mechanisms governing HIF-1α in ILC3s and other innate RORγt+ cells during in vivo infections are poorly understood. In our study, transgenic mice with specific Hif-1a gene inactivation in innate RORγt+ cells (RAG1KO HIF-1α▵Rorc) exhibit more severe colitis following Citrobacter rodentium infection, primarily due to the inability to upregulate IL-22. We find that HIF-1α▵Rorc mice have impaired IL-22 production in ILC3s, while non-ILC3 innate RORγt+ cells, also capable of producing IL-22, remain unaffected. Furthermore, we show that IL-18, induced by Toll-like receptor 2, selectively triggers IL-22 in ILC3s by transcriptionally upregulating HIF-1α, revealing an oxygen-independent regulatory pathway. Our results highlight that, during late-stage C. rodentium infection, IL-18 induction in the colon promotes IL-22 through HIF-1α in ILC3s, which is crucial for protection against this pathogen.


Induction of metamorphosis in the sea urchin Holopneustes purpurascens by a metabolite complex from the algal host Delisea pulchra.
Most benthic invertebrates have complex life cycles with planktonic larvae that return to the substratum to settle and metamorphose into a benthic stage. Although naturally produced chemical cues have long been thought to be important for the settlement or metamorphosis of invertebrate larvae, few ecologically relevant chemical cues have been clearly identified. The marine echinoid Holopneustes purpurascens has a complex life cycle, with a planktonic, nonfeeding dispersive larva that metamorphoses into a benthic stage that lives in the canopy of subtidal benthic algae such as the red alga Delisea pulchra and the kelp Ecklonia radiata. Recently recruited juveniles are found primarily on D. pulchra, and we hypothesized that this was in response to a chemical cue produced by this alga. Competent larvae metamorphosed in the presence of D. pulchra, or seawater surrounding this alga, but not in response to the presence of E. radiata or its extracts. A cue for metamorphosis was isolated and characterized from D. pulchra and found to be a water-soluble complex of the sugar floridoside and isethionic acid in a 1:1 molar ratio. The floridoside-isethionic acid complex also triggered settlement in H. purpurascens; however, this response was less specific than metamorphosis and was reversible. Larvae of H. purpurascens also metamorphosed in the presence of several other species of red, but not brown or green, algae from their habitat. Floridoside is found only in red algae, suggesting that the floridoside-isethionic acid complex may be acting as a cue for metamorphosis in other red algae as well as in D. pulchra.


Functional hydrophobin-coating of thermally hydrocarbonized porous silicon microparticles.
Porous silicon (PSi) particles have been widely used in modulating the dissolution rate of various types of drugs loaded within its mesopores. This material can be surface treated in order to vary its hydrophobicity and several other properties, such as drug loading degree and release rate. Hydrophobins are a family of self-assembling proteins of fungal origin which have the ability to form layers on hydrophobic materials. This type of protein layer can modify the characteristics and control the binding properties of the surface on which it assembles. In this study, we have developed a procedure to coat thermally hydrocarbonized-PSi microparticles with hydrophobin II (HFBII) in order to modify the particles' hydrophobicity and to improve their biocompatibility, while maintaining intact the advantageous drug releasing properties of the PSi. The HFBII content adsorbed onto the particles was successfully quantified by a protein assay. Drug dissolution and permeation across Caco-2 cell monolayers were also conducted, together with viability studies in AGS, Caco-2 and HT-29 cells. The characterization and coating stability assessment showed that the HFBII-coating desorbs partially from the particles' surface as the pH increases. The HFBII coating also improved the biocompatibility of the particles without compromising the enhanced drug permeation or release.


Esthetic Rehabilitation of Primary Anterior Teeth using Temporization Material: A Novel Approach.
Pediatric dentists should always aim for esthetic and functional rehabilitation of decayed/traumatized primary teeth. The most common method for restoring such teeth involves the use of "strip crowns" with composites, while the recent trend is toward using other extracoronal restorations including preve-neered stainless steel crowns and zirconia crowns. All these restorative options have shown good success rates, but also have some limitations. This case series depicts novel clinical technique of using a temporization material for full-coronal restoration(s) in primary anterior teeth. This included the chair-side custom fabrication of full-coronal restoration using temporization material, which has resulted in good immediate esthetics and might be a cost-effective alternative for restoring primary anterior teeth in future. Gugnani N, Pandit IK, Gupta M, Nagpal J. Esthetic Rehabilitation of Primary Anterior Teeth using Temporization Material: A Novel Approach. Int J Clin Pediatr Dent 2017;10(1):111-114.


Cross-sectional relations of digital vascular function to cardiovascular risk factors in the Framingham Heart Study.
Digital pulse amplitude augmentation in response to hyperemia is a novel measure of peripheral vasodilator function that depends partially on endothelium-derived nitric oxide. Baseline digital pulse amplitude reflects local peripheral arterial tone. The relation of digital pulse amplitude and digital hyperemic response to cardiovascular risk factors in the community is unknown. Using a fingertip peripheral arterial tonometry (PAT) device, we measured digital pulse amplitude in Framingham Third Generation Cohort participants (n=1957; mean age, 40+/-9 years; 49% women) at baseline and in 30-second intervals for 4 minutes during reactive hyperemia induced by 5-minute forearm cuff occlusion. To evaluate the vascular response in relation to baseline, adjusting for systemic effects and skewed data, we expressed the hyperemic response (called the PAT ratio) as the natural logarithm of the ratio of postdeflation to baseline pulse amplitude in the hyperemic finger divided by the same ratio in the contralateral finger that served as control. The relation of the PAT ratio to cardiovascular risk factors was strongest in the 90- to 120-second postdeflation interval (overall model R(2)=0.159). In stepwise multivariable linear regression models, male sex, body mass index, ratio of total to high-density lipoprotein cholesterol, diabetes mellitus, smoking, and lipid-lowering treatment were inversely related to PAT ratio, whereas increasing age was positively related to PAT ratio (all P<0.01). Reactive hyperemia produced a time-dependent increase in fingertip pulse amplitude. Digital vasodilator function is related to multiple traditional and metabolic cardiovascular risk factors. Our findings support further investigations to define the clinical utility and predictive value of digital pulse amplitude.


Modeling of Human Hepatic and Gastrointestinal Ethanol Metabolism with Kinetic-Mechanism-Based Full-Rate Equations of the Component Alcohol Dehydrogenase Isozymes and Allozymes.
Alcohol dehydrogenase (ADH) is the principal enzyme responsible for the metabolism of ethanol. Human ADH constitutes a complex family of isozymes and allozymes with striking variation in kinetic properties and tissue distribution. The liver and the gastrointestinal tract are the major sites for first-pass metabolism (FPM). The quantitative contributions of ADH isozymes and ethnically distinct allozymes to cellular ethanol metabolism remain poorly understood. To address this issue, kinetic mechanism and the steady-state full-rate equations for recombinant human class I ADH1A, ADH1B (including allozymes ADH1B1, ADH1B2, and ADH1B3), ADH1C (including allozymes ADH1C1 and ADH1C2), class II ADH2, and class IV ADH4 were determined by initial velocity, product inhibition, and dead-end inhibition experiments in 0.1 M sodium phosphate at pH 7.5 and 25 °C. Models of the hepatic and gastrointestinal metabolisms of ethanol were constructed by linear combination of the numerical full-rate equations of the component isozymes and allozymes in target organs. The organ simulations indicate that in homozygous ADH1B*1/*1 livers, a representative genotype among ethnically distinct populations due to high prevalence of the allele, major contributors at 1 to 10 mM ethanol are ADH1B1 (45% to 24%) and the ADH1C allozymes (54% to 40%). The simulated activities at 1 to 50 mM ethanol for the gastrointestinal tract (total mucosae of ADH1C*1/*1-ADH4 stomach and the ADH1C*1/*1-ADH2 duodenum and jejunum) account for 0.68%-0.76% of that for the ADH1B*1/*1-ADH1C*1/*1 liver, suggesting gastrointestinal tract plays a relatively minor role in the human FPM of ethanol. Based on the flow-limited sinusoidal perfusion model, the simulated hepatic Kmapp, Vmaxapp, and Ci at a 95% clearance of ethanol for ADH1B*1/*1-ADH1C*1/*1 livers are compatible to that documented in hepatic vein catheterization and pharmacokinetic studies with humans that controlled for the genotypes. The model simulations suggest that slightly higher or similar ethanol elimination rates for ADH1B*2/*2 and ADH1B*3/*3 individuals compared with those for ADH1B*1/*1 individuals may result from higher hepatocellular acetaldehyde.


Laparoscopic gastric surgery in a Japanese institution: analysis of the initial 100 procedures.
Although endoscopic surgical procedures are popular in various fields, reports on its use in gastric surgical procedures are limited. This study was designed to review our initial experience with laparoscopic gastric surgical techniques to evaluate indications and surgical results. We undertook a retrospective analysis of 100 patients (66 men and 34 women, mean age 63 years) who underwent laparoscopic gastric surgical procedures between 1995 and 2001. Procedures performed were distal gastrectomy (n = 76), wedge resection (n = 20), and intragastric surgical procedures (n = 4). Patients were divided into two groups according to the date of the procedure, from the earliest to the most recent. There were 85 patients with gastric cancers, 14 submucosal tumors, and 1 duodenal ulcer. In 8 cases conversion was made to an open surgical procedure. Operation times required for distal gastrectomy, wedge resection, and intragastric surgical procedures were 330 +/- 69, 144 +/- 34, and 298 +/- 106 min, and blood loss was 354 +/- 251, 56 +/- 94, and 33 +/- 58 g, respectively. Complications included transient anastomotic stenosis (n = 5), leakage (n = 4), and bleeding (n = 1) after distal gastrectomy, and bleeding (n = 1) after intragastric surgical procedures. There were no complications after wedge resection. Comparing the first and second halves of the series, the percentage of distal gastrectomy significantly increased from 66% to 86% (p = 0.02) and the number of dissected lymph nodes at this procedure increased from 20 +/- 13 to 33 +/- 17 (p < 0.01). Laparoscopic gastric surgical procedures are safe and feasible for early gastric cancers and submucosal tumors. Technical advances in lymph node dissection have made distal gastrectomy a leading and increasingly popular laparoscopic procedure for early gastric cancer.


Organ weight changes associated with body mass index determined from a medical autopsy population.
Existing organ weight charts used by pathologists for patients undergoing medical autopsy do not illustrate the effect of obesity and age on organ weights among a general population of older individuals with multiple comorbidities. We retrospectively reviewed 300 medical autopsy reports to extract data to analyze the effect of obesity and age on organ weights. In both men and women, there were statistically significant increases in organ weights with body mass index (BMI) but decreases with age for liver, spleen, and kidneys. In men, increased age was associated with increased left ventricular wall thickness, whereas increased BMI was associated with increased heart weight. In women, only BMI was associated with changes in all 3 anatomic cardiac parameters (heart weight and thickness of the right and left ventricular walls). Age effects were not observed for heart parameters in women. Thyroid weight increased with BMI in men but not in women. The findings demonstrate changes in organ weights/sizes with obesity and age in a population of patients with multiple comorbidities. The differential effects of age and BMI on the heart between men and women raise the possibility that increased BMI in women may have a greater impact on cardiovascular causes of death than that in men.


The osmotic demyelination syndrome: the resilience of thalamic neurons is verified with transmission electron microscopy.
The development of a murine model of osmotic demyelinating syndrome (ODS) allowed to study changes incurred in extrapontine zones of the CNS and featured neuron and glial cell changes in the relay thalamic ventral posterolateral (VPL) and ventral posteromedial (VPM) nuclei before, during and after ODS induction, and characterized without immune response. There, the neuron Wallerian-type deteriorations were verified with fine structure modifications of the neuron cell body, including some nucleus topology and its nucleolus changes. Morphologic analyses showed a transient stoppage of transcriptional activities while myelinated axons in the surrounding neuropil incurred diverse damages, previously reported. Even though the regional thalamus myelin deterioration was clearly recognized with light microscopy 248 h after osmotic recovery of ODS, ultrastructure analyses demonstrated that, at that time, the same damaged parenchyma regions contained nerve cell bodies that have already reactivated nucleus transcriptions and neuroplasm translations because peculiar accumulations of fibro-granular materials, similar to those detected in restored ODS astrocytes, were revealed in these restructuring nerve cell bodies. Their aspects suggested to be accumulations of ribonucleoproteins. The findings suggested that progressive neural function's recovery in the murine model could imitate some aspects of human ODS recovery cases.


[Regenerative capacity in multiple sclerosis].
Disability in multiple sclerosis is related to demyelination and axonal injury. Remyelination has been shown to occur in the different forms of the disease. In addition to restoring nerve conduction, myelin repair has a major role in preventing neurodegeneration. However, although sometimes extensive, remyelination is usually insufficient. Our understanding of the cellular and molecular mechanisms underlying the success or failure of myelin repair has benefited from lesion analysis in multiple sclerosis, and from in vivo and in vitro models of myelination and remyelination. This also provides potential targets for therapeutic intervention aimed at promoting endogenous remyelination.


Effects of a phytoestrogen-containing soy extract on the growth-inhibitory activity of ICI 182 780 in an experimental model of estrogen-dependent breast cancer.
The study reported here was designed to determine whether a phytoestrogen-containing soy extract (SSE) could negate/overwhelm the inhibitory effects of ICI 182 780 on the growth of estrogen-sustained human breast cancer xenografts (MCF-7), in ovariectomized athymic mice. As expected, estradiol-supplemented tumors did not grow over the study period in ICI 182 780-treated females; concomitant administration of 50 mg/kg per day SSE slightly potentiated the inhibitory activity of the drug, while at 100 mg/kg per day, SSE partially negated ICI 182 780 activity. In keeping with these in vivo outcomes, we observed that the level of cyclin D1 (and progesterone receptor) in MCF-7 xenografts was considerably reduced by ICI 182 780, an effect enhanced by concomitant treatment with 50 SSE, but reduced by the higher dosage (i.e. 100 mg/kg per day). Thrombospondin-1 (TSP-1) and kallikrein 6 (KLK6) levels were also reduced following ICI 182 780, although to a lesser degree; again, combined anti-estrogen and SSE produced a dose-dependent regulation in TSP-1 and KLK6 tumor level, with a further reduction in the mRNA gene expression at 50 SSE (compared with ICI 182 780) and a partial reversion of the drug-induced down-regulation at 100 mg/kg per day. No modulation was detected in the serum concentration of IGF-1 (a potent mitogen for estrogen receptor-positive breast cancer cell lines) either upon treatment with ICI 182 780 or concomitant administration of the anti-estrogen with SSE. In conclusion, results from this study raise concerns about the consumption of isoflavone supplements in conjunction with ICI 182 780 therapy, in postmenopausal women with estrogen-dependent breast cancer.


[Skeletal musculature of rats during modeling of the physiological effects of weightlessness (morphological study)].
In the experiment rats were suspended in the head-down position (at -15 to -20 degrees) for 21 days. To study the effect of this exposure on skeletal muscles of their fore- and hindlimbs, morphological and histochemical methods were used. Changes in the hindlimbs were similar to those seen in real weightlessness of similar duration. The two antigravitational muscles - soleus and gastrocnemius muscles - showed greatest changes, i.e. atrophic and metabolic shifts. The biceps brachii muscle that changes insignificantly in the weightless state developed a destructive process (with degradation of myofibers) that was later replaced with a reparative process. The changes in this muscle are assumed to be associated with hemodynamic disorders. It is recognized that the method of rat suspension is adequate for a ground-based study of morphological effects of weightlessness on hindlimbs and is unacceptable for that of forelimbs where the exposure induces microcirculation disorders.


How then should we die?: California's "Death with Dignity" Act.
The cultural significance of recent changes in medicine and advances in biotechnology can hardly be overstated. Such have stirred fresh interest in the moral foundations of ethical decision-making and lively debate has ensued as well over the basis of human dignity. Largely divorced from the distinctive moral and ethical commitments that once informed and directed medical practice, modern secular notions of bioethics collapse frequently into human philosophical models of rights and justice. The project of Western medicine now continues within the cultural framework of a radical postmodern agenda that calls for the critical deconstruction and absolute relativizing of all knowledge, and the thorough secularization of the public square. Truth, once understood as a fixed expression of a fundamental reality, has been eschewed in favor of personal preference, subjective experience, private interpretation, and radical perspectivism. Perhaps the greatest challenge facing contemporary culture is that of arriving at a clear and convincing consensus on what constitutes moral surety as well as agreement on what knowledge can serve as an adequate foundation for living the moral life. Having abandoned the reality of divine involvement in the creation and sustenance of human life, contemporary culture now toys with what it means to be human without God. Increasingly popular is the view that whether one possesses dignity or not turns on the question of suffering. Under modern parlance it is simply undignified to suffer. Suffering, somehow, is believed to reduce a person to a state that is incompatible with dignity. Therefore, it should come as no surprise that one of the most pressing social issues today involves the effort to legalize physician-assisted suicide--a project based upon the view that people ought to "die with dignity." In California this view is embodied in Assembly Bill 1592, The "Death with Dignity Act." It is intended to establish California as the second state in the United States to legalize physician-assisted suicide. Patterned closely after Oregon's groundbreaking law, the California bill provides for a terminally ill patient to end his or her own life when certain conditions exist. The purpose of this paper is to review the historical development of California Assembly Bill 1592. An evaluation of both the content of this bill and the process by which it was introduced to the California State Assembly will be offered. The student of bioethics will here find that the usual arguments in support of physician-assisted suicide in the United States, as well as euthanasia in the Netherlands, have been employed in support of AB1592; namely suffering and autonomy. Given the rather predictable pattern of argument advanced by advocates of physician-assisted suicide and euthanasia, this discussion will prove useful for those who wish to offer an informed and sensitive Christian response to similar legislative efforts in other states and countries.


Prolonged lung retention of iodine-123-MIBG in diabetic patients.
Iodine-123-MIBG has been introduced as a biochemical marker in assessing pulmonary endothelial cell integrity and myocardial beta-adrenergic sympathetic innervation. The aim of this study was to evaluate 123I-MIBG lung uptake in diabetic patients with and without coronary artery disease. Forty-four nonsmoking patients with normal respiratory function tests were included: 12 diabetics, 11 diabetics with coronary artery disease, 14 nondiabetic patients with coronary artery disease and 7 age-matched controls were imaged with 123I-MIBG and 201Tl scintigraphy. The lung retention of 123I-MIBG, cardiac sympathetic innervation (heart-to-upper mediastinum ratio of 123I-MIBG) and 201Tl lung-to-heart ratio were determined in all cases. In diabetics with coronary artery disease, significantly prolonged lung retention and decreased cardiac uptake of 123I-MIBG were found. The lung retention of 123I-MIBG was inversely correlated with the heart-to-upper mediastinum ratio in this group. Lung-to-heart ratios of 201Tl were significantly increased in patients with coronary artery disease but there was no significant difference between diabetics and nondiabetics. Prolonged lung retention of 123I-MIBG was associated with decreased cardiac sympathetic innervation in diabetic coronary artery disease patients. It seems that passive pulmonary congestion or cardiac dysfunction itself did not influence 123I-MIBG lung uptake. Increased lung extraction of 123I-MIBG is highly suggestive of ongoing pulmonary endothelial dysfunction together with ischemic events in diabetics.


Long-term live-cell imaging of mammalian preimplantation development and derivation process of pluripotent stem cells from the embryos.
Mammalian fertilization is a process in which two highly specialized haploid gametes unite and endow totipotency to the resulting diploid zygote. This is followed by cell proliferation and the onset of differentiation during the brief period leading up to implantation. In these processes, a number of cellular components and structures are regulated spatially and temporally, as seen in repeated cell division, cell cycle progression, and epigenetic reprogramming. In mammals, the numbers of oocytes and embryos that can be collected are very limited. Therefore, analyses of molecular mechanisms are hampered because of difficulties in conducting biochemical analyses on such limited material. Furthermore, immunostaining methods require cell fixation and are insufficient for understanding ontogeny, because the processes observed in fertilization and early embryonic development progress in time-dependent manners and each phenomenon is connected with others by cause-and-effect relationships. Consequently, it is important to develop an experimental system that enables molecular imaging without affecting embryonic development. To achieve the above advantages, especially retrospective and prospective analyses, we have established a live-cell imaging system that enables observations under minimally invasive conditions. Using this approach, we have succeeded in visualizing and predicting the developmental potential of embryos after various perturbations. We also succeeded in imaging embryonic stem (ES) cell derivation in natural conditions. In this review, we describe a brief history of embryonic imaging and detailed protocols. We also discuss promising aspects of imaging in the fields of developmental and stem cell biology.


Temporal invariance and clonal uniformity of brain and cerebrospinal IgG, IgA, and IgM in multiple sclerosis.
Elevated cerebrospinal fluid (CSF) IgG and oligoclonal IgG bands on electrophoresis are valuable clinical markers for B cell proliferation in the brains of patients with multiple sclerosis (MS). Using two-dimensional electrophoresis, (2DE) we have established that the humoral immune response in MS brain is characterized by finite clonal complexity for the major Ig classes. An important question is whether this immune response is clonally stable or varies with time, related to the development of new lesions and random entry of B cells into the MS brain. To investigate this, we performed serial electrophoretic studies on CSF obtained from 19 patients with MS; the intervals ranged from 7 to 12 yr, with a mean of 8 yr. These analyses included studies of IgG, IgA, and IgM, and revealed that the humoral immune response in MS is clonally stable over long periods. Spontaneous fluctuations or reduction in CSF IgG levels by drugs did not qualitatively affect B cell clonal proliferation in MS brain, in that dominant bands and spots were not obliterated. It has been asserted that IgG synthesis in MS is nonsense antibody because the spectotypes of IgG isolated from different regions of MS brains differ. Factors other than clonal heterogeneity could account for differences found using one-dimensional analysis. B cell clonal products resolve into unique and well-resolved spots by 2DE; the method is uniquely suitable for analysis of restricted immune responses. Therefore, Ig were isolated from 11 regions of three MS brains and the 2DE patterns were compared. The similarity of the 2DE patterns indicate unequivocally that major clones are distributed uniformly although some clones are more prominent in some brain areas. IgA and IgM isolated from the same areas also showed similar patterns. Furthermore, the patterns of light and heavy chains in brain regions differed from serum but were similar to the autologous CSF, providing new evidence that CSF IgG in MS derives from synthesis in situ. Our results indicate that, once initiated, B cell clonal proliferation persists indefinitely and is little altered qualitatively at a clonal level over time, even when CSF IgG levels change or are altered by drugs. Our results are consistent with allotype and idiotype analysis of Ig production in MS and conflict with nonsense antibody proposals of the origin and nature of in situ synthesized Ig in MS.


Expression of neurotrophin-4/5 in Streptococcus pneumoniae meningitis.
This study aims to investigate the expression of neurotrophin-4/5 (NT-4/5) in the pallium and hippocampus in juvenile rats with intraventricular injection of Streptococcus pneumoniae. We used 40 SPF SD rats (3 weeks old, regardless of gender) in this study. We drew 50 μL cerebrospinal fluid, and then, we injected 50 μL normal saline and S. pneumoniae suspension (10(8) CFU/mL) in the brain pool (normal control group and infection group, respectively). After 24 h, the cerebrospinal fluid was collected for bacterial culture and white blood cell count. The immunohistochemical staining was conducted at the day 1, 2, and 5, and the expression of NT-4/5 in rat brain tissue was observed. Compared with the normal control group, NT-4/5 expression in the pallium and hippocampus of rats in the 24-h infection group was significantly increased (both P < 0.05). NT-4/5 expression in the pallium and hippocampus in the 5-day infection group was significantly lower than that in the 24-h infection group (P < 0.05), and they were significantly higher than the normal control group (P < 0.05). After intraventricular injection of S. pneumoniae, the expression of NT-4/5 in the pallium and hippocampus in juvenile rats was increased, especially during early disease course.


Development of genodynamic metrics for exploring the biophysics of DNA polymorphisms.
Single nucleotide polymorphisms (SNPs) represent an important type of dynamic sites within the human genome. These common variants often locally correlate within more complex multi-SNP haploblocks that are maintained throughout generations in a stable population. Information encoded in the structure of SNPs and SNP haploblock variation can be characterized through a normalized information content metric. Genodynamics is being developed as the analogous "thermodynamics" characterizing the state variables for genomic populations that are stable under stochastic environmental stresses. Since living systems have not been found to develop in the absence of environmental influences, this paper describes the analogous genomic free energy metrics in a given environment. SNP haploblocks were constructed by Haploview v4.2 for five chromosomes from phase III HapMap data, and the genomic state variables for each chromosome were calculated. An in silico analysis was performed on SNP haploblocks with the lowest genomic energy measures. Highly favorable genomic energy measures were found to correlate with highly conserved SNP haploblocks. Moreover, the most conserved haploblocks were associated with an evolutionarily conserved regulatory element and domain.


Patchouli alcohol ameliorates dextran sodium sulfate-induced experimental colitis and suppresses tryptophan catabolism.
Despite the increased morbidity of ulcerative colitis (UC) in recent years, available treatments remain unsatisfactory. Pogostemon cablin has been widely applied to treat a variety of gastrointestinal disorders in clinic for centuries, in which patchouli alcohol (PA, C15H26O) has been identified as the major active component. This study attempted to determine the bioactivity of PA on dextran sulfate sodium (DSS)-induced mice colitis and clarify the mechanism of action. Acute colitis was induced in mice by 3% DSS for 7 days. The mice were then given PA (10, 20 and 40mg/kg) or sulfasalazine (SASP, 200mg/kg) as positive control via oral administration for 7 days. At the end of study, animals were sacrificed and samples were collected for pathological and other analysis. In addition, a metabolite profiling and a targeted metabolite analysis, based on the Ultra-Performance Liquid Chromatography coupled with mass spectrometry (UPLC-MS) approach, were performed to characterize the metabolic changes in plasma. The results revealed that PA significantly reduced the disease activity index (DAI) and ameliorated the colonic injury of DSS mice. The levels of colonic MPO and cytokines involving TNF-α, IFN-γ, IL-1β, IL-6, IL-4 and IL-10 also declined. Furthermore, PA improved the intestinal epithelial barrier by enhancing the level of colonic expression of the tight junction (TJ) proteins, for instance ZO-1, ZO-2, claudin-1 and occludin, and by elevating the levels of mucin-1 and mucin-2 mRNA. The study also demonstrated that PA inhibited the DSS-induced cell death signaling by modulating the apoptosis related Bax and Bcl-2 proteins and down-regulating the necroptosis related RIP3 and MLKL proteins. By comparison, up-regulation of IDO-1 and TPH-1 protein expression in DSS group was suppressed by PA, which was in line with the declined levels of kynurenine (Kyn) and 5-hydroxytryptophan (5-HTP) in plasma. The therapeutic effect of PA was evidently reduced when Kyn was given to mice. In summary, the study successfully demonstrated that PA ameliorated DSS-induced mice acute colitis by suppressing inflammation, maintaining the integrity of intestinal epithelial barrier, inhibiting cell death signaling, and suppressing tryptophan catabolism. The results provided valuable information and guidance for using PA in treatment of UC.


Combining smoking information and molecular markers improves prognostication in patients with urothelial carcinoma of the bladder.
Tissue-based markers improve the accuracy of prediction models in urothelial carcinoma of the bladder (UCB). Current smoking status and cumulative exposure also affect outcomes. To evaluate whether the combination of molecular markers and smoking features further improved the prognostication of patients who underwent radical cystectomy (RC) for UCB. A total of 588 patients underwent RC and bilateral lymphadenectomy for UCB from 1995 to 2005. Immunohistochemistry for p53, p21, pRB, p27, Ki-67, and survivin was performed on tissue microarrays from the RC specimen. Smoking features were routinely assessed at diagnosis. Multivariable Cox regression models assessed time to disease recurrence and cancer-specific mortality. Of the 588 patients, 128 were never (22%), 283 former (48%), and 177 current smokers (30%). In total, 227 patients experienced disease recurrence, whereas 190 died of UCB. Smoking status was independently associated with both outcomes (hazard ratio [HR] = 1.48 and 2.62, for former and current vs. never smokers, respectively, P<0.001). All markers were significantly associated with both outcomes (P<0.05) except for survivin. The combination of the 4 cell cycle markers p53, p21, pRB, and p27 increased the discrimination of clinicopathologic model for former and current vs. never smokers with c-indices 0.779 and 0.780, respectively (base model c-indices of 0.741 and 0.740 for former and current vs. never smokers, respectively). The further addition of smoking features and biomarker status improved the discrimination of the model (c-indices of 0.783 and 0.786 for former and current vs. never smokers, respectively). We confirmed that smoking information and tissue markers status improve prognostication of UCB outcomes after RC; the combination of both reaching the highest level of discrimination.


Role of renal metabolism and excretion in 5-nitrofuran-induced uroepithelial cancer in the rat.
5-Nitrofurans have been used in the study of chemical carcinogenesis. There is substantial evidence that N-[4-(5-nitro-2-furyl)-2-thiazolyl] formamide (FANFT) is deformylated to 2-amino-4-(5-nitro-2-furyl)thiazole (ANFT) in the process of FANFT-induced bladder cancer. Paradoxically, ANFT is less potent as a uroepithelial carcinogen than FANFT when fed to rats. Feeding aspirin with FANFT to rats decreases the incidence of bladder cancer. Isolated kidneys were perfused with 5-nitrofurans to determine renal clearances and whether aspirin acts to decrease urinary excretion of the carcinogen. In FANFT-perfused kidneys, FANFT was deformylated to ANFT and excreted (1.06 +/- 0.22 nmol/min) at a rate eightfold higher than excretion of FANFT. In kidneys perfused with equimolar ANFT, excretion of ANFT was 0.25 +/- 0.05 nmol/min, which suggests a coupling of renal deformylation of FANFT to excretion of ANFT in FANFT-perfused kidneys. Neither aspirin nor probenecid altered the urinary excretion or half-life of FANFT or ANFT. In rats fed 0.2% FANFT as part of their diet, coadministration of aspirin (0.5%) increased urinary excretion of ANFT during a 12-wk feeding study, which suggests decreased tissue binding or metabolism of ANFT. Kidney perfusion with acetylated ANFT (NFTA), a much less potent uroepithelial carcinogen, resulted in no ANFT excretion or accumulation, which indicates the specificity of renal deformylase. Renal deformylase activity was found in broken cell preparations of rat and human kidney. These data describe a unique renal metabolic/excretory coupling for these compounds that appears to explain the differential carcinogenic potential of the 5-nitrofurans tested. These results are consistent with the hypothesis that aspirin decreases activation of ANFT by inhibiting prostaglandin H synthase.


Determination of electroosmotic flow in nonaqueous capillary electrophoresis.
Mobility of the electroosmotic flow (mu(EOF)) in fused-silica capillaries strongly depends on the nature of the background electrolyte. In this study, 27 solvent systems were investigated, namely water, methanol, ethanol, 2-propanol, 1-butanol, acetonitrile (MeCN), formamide, N-methylformamide (NMF), N,N-dimethylformamide and dimethyl sulfoxyde, as well as 8 hydroorganic and 9 organic mixtures. For each system, six mu(EOF) were determined at a different ionic strength in basic conditions, and an absolute electroosmotic flow mobility (mu(EOF,0)) was extrapolated according to the Debye-Huckel Onsager model. The obtained mu(EOF,0) values were correlated with the solvent's relative permittivity (epsilon) and viscosity (eta). A good correlation (r2=0.867) between mu(EOF,0) and the solvent's epsilon/eta ratio was demonstrated, except for two solvents (MeCN and NMF). Furthermore, the donor number (DN) of a solvent took into account the possible zeta potential modification in the electric double layer near the capillary wall. Consequently, the relationship between mu(EOF,0) and epsilon/(eta x DN) was superior, with a r2 of 0.943 for 10 pure solvents.


Stronger findings from mass spectral data through multi-peak modeling.
Mass spectrometry-based metabolomic analysis depends upon the identification of spectral peaks by their mass and retention time. Statistical analysis that follows the identification currently relies on one main peak of each compound. However, a compound present in the sample typically produces several spectral peaks due to its isotopic properties and the ionization process of the mass spectrometer device. In this work, we investigate the extent to which these additional peaks can be used to increase the statistical strength of differential analysis. We present a Bayesian approach for integrating data of multiple detected peaks that come from one compound. We demonstrate the approach through a simulated experiment and validate it on ultra performance liquid chromatography-mass spectrometry (UPLC-MS) experiments for metabolomics and lipidomics. Peaks that are likely to be associated with one compound can be clustered by the similarity of their chromatographic shape. Changes of concentration between sample groups can be inferred more accurately when multiple peaks are available. When the sample-size is limited, the proposed multi-peak approach improves the accuracy at inferring covariate effects. An R implementation and data are available at http://research.ics.aalto.fi/mi/software/peakANOVA/.


Molecular mobility in biodegradable poly(ε-caprolactone)/poly(hydroxyethyl acrylate) networks.
Poly(ε-caprolactone)/poly(hydroxyethyl acrylate) networks have been investigated by thermally stimulated depolarization currents (TSDC) and differential scanning calorimetry (DSC). The introduction of hydrophilic units (HEA) in the system aiming at tailoring the hydrophilicity of the system results in a series of copolymer networks with microphase separation into hydrophobic/hydrophilic domains. Polycaprolactone (PCL) crystallization is prevented by the topological constraints HEA units imposed in such heterogeneous domains. Moreover, the mobility of the amorphous PCL chains is enhanced as revealed by the main relaxation process which becomes faster. The glass transition of PHEA-rich domains shifts to lower temperatures, as the total amount of PCL in the copolymer increases, due to the presence of PCL units within the same region. The behaviour of the copolymer networks swollen with different content of water has been investigated to analyze the interaction between water molecules and hydrophobic/hydrophilic domains and provide further insights into the molecular structure of the system.


Air-Puff-Induced Dynamics of Ocular Components Measured with Optical Biometry.
To analyze the dynamics of all optical components of the eye and the behavior of the eyeball under air-puff conditions in vivo. To determine the impact of the intraocular pressure (IOP) on the air-puff-induced deformation of the eye. Twenty eyes of 20 healthy subjects were included in this study. The dynamics of the ocular components, such as the cornea, the crystalline lens, and the retina, was measured by a prototype swept source optical coherence tomography biometer integrated with the air-puff system. The system allows to acquire a series of axial scans at the same location as a function of time with no transverse scanning. Several parameters were extracted from optical coherence tomography data. The IOP was measured using a Goldmann applanation tonometry. The measurements of the eyes were performed before and 2 hours after administration of IOP-reducing drops, namely, 0.2 % brimonidine tartrate. There is a statistically significant correlation of corneal thickness, vitreous depth, and eye length with IOP. The deformation amplitudes of the cornea and the crystalline lens are inversely proportional to the IOP, but statistical significance is achieved only for the cornea. The crystalline lens is displaced without compression, and the return has the form of wobbling. The reduction of IOP level induces corresponding changes in the extracted parameters. Optical biometry combined with air puff provides comprehensive information on the in vivo behavior of all ocular components, including the crystalline lens. Measurement of the axial length dynamics of during deformation enables correcting the deformation for eye retraction.


Expression analysis of extracellular proteins from Phanerochaete chrysosporium grown on different liquid and solid substrates.
White-rot fungi secret a large number of hydrolytic and oxidative enzymes for degradation of lignocellulosic material. The sequencing of the genome of the white-rot fungus Phanerochaete chrysosporium has facilitated the characterization of its complete extracellular proteome. P. chrysosporium was grown on liquid medium, containing glucose, cellulose or wood chips as the carbon source, and also in solid substrate fermentation bags. For liquid-grown cultures, the extracellular protein fraction was separated by 2D gel electrophoresis. Protein spots were analysed by in-gel digestion and liquid chromatography (LC)/MS/MS. A total of 18 additional protein spots from the 2D gels yielded hits from blast searches. From solid substrate cultures in which the fungus was grown in bags, the proteins were resolved by SDS-PAGE, subjected to in-gel digestion and then identified by LC/MS/MS. An additional 16 proteins yielded hits on blast searches. Enzymes involved in cellulose, hemicellulose, lignin and protein degradation were identified. Expression patterns were very similar between cellulose-grown cultures and wood-grown cultures. In addition to enzymes which act on lignocellulosic material, proteases were also found, indicating the need of fungi to scavenge for nitrogen in wood.


Expansion and Activation of CD103(+) Dendritic Cell Progenitors at the Tumor Site Enhances Tumor Responses to Therapeutic PD-L1 and BRAF Inhibition.
Large numbers of melanoma lesions develop resistance to targeted inhibition of mutant BRAF or fail to respond to checkpoint blockade. We explored whether modulation of intratumoral antigen-presenting cells (APCs) could increase responses to these therapies. Using mouse melanoma models, we found that CD103(+) dendritic cells (DCs) were the only APCs transporting intact antigens to the lymph nodes and priming tumor-specific CD8(+) T cells. CD103(+) DCs were required to promote anti-tumoral effects upon blockade of the checkpoint ligand PD-L1; however, PD-L1 inhibition only led to partial responses. Systemic administration of the growth factor FLT3L followed by intratumoral poly I:C injections expanded and activated CD103(+) DC progenitors in the tumor, enhancing responses to BRAF and PD-L1 blockade and protecting mice from tumor rechallenge. Thus, the paucity of activated CD103(+) DCs in tumors limits checkpoint-blockade efficacy and combined FLT3L and poly I:C therapy can enhance tumor responses to checkpoint and BRAF blockade.


The predominant WT1 isoform (+KTS) encodes a DNA-binding protein targeting the planar cell polarity gene Scribble in renal podocytes.
WT1 encodes a tumor suppressor first identified by its inactivation in Wilms' Tumor. Although one WT1 splicing variant encodes a well-characterized zinc finger transcription factor, little is known about the function of the most prevalent WT1 isoform, whose DNA binding domain is disrupted by a three-amino acid (KTS) insertion. Using cells that conditionally express WT1(+KTS), we undertook a genome-wide chromatin immunoprecipitation and cloning analysis to identify candidate WT1(+KTS)-regulated promoters. We identified the planar cell polarity gene Scribble (SCRB) as the first WT1(+KTS) target gene in podocytes of the kidney. WT1 and SCRB expression patterns overlap precisely in developing renal glomeruli of mice, and WT1(+KTS) binds to a 33-nucleotide region within the Scribble promoter in mouse and human cell lines and kidneys. Together, our results support a role for the predominant WT1(+KTS) isoform in transcriptional regulation and suggest a link between the WT1-dependent tumor suppressor pathway and a key component of the planar cell polarity pathway.


Chaos synchronization and communication in closed-loop semiconductor lasers subject to common chaotic phase-modulated feedback.
We propose and demonstrate a closed-loop chaos system composed of external-cavity semiconductor lasers subject to common chaotic phase-modulated optical feedback (CCPMOF). The efficient-bandwidth and time-delay signature (TDS) characteristics of the chaotic carrier, the properties of chaos synchronization, as well as the performance and security of chaos communication are systematically investigated. The numerical results demonstrate that wideband chaotic carrier with effective TDS suppression can be easily obtained, high-quality chaos synchronization with considerable mismatch robustness, frequency detuning tolerance, and phase fluctuation tolerance can be achieved in a wide operation range, and high-speed chaos communication is available. With respect to the conventional closed-loop systems, the bandwidth and complexity of chaotic carrier is greatly enhanced, and the performances of chaos synchronization and communication are obviously improved.


Agenda for patient-centered care research in Brazil.
Patient-centered care is an incipient movement and its practice still faces obstacles in the Brazilian health system, where it is not extensively identified as healthcare quality realm. Thus, this paper aims to establish a patient/person-centered care research agenda to support its implementation in the country's healthcare services. A panel was held with nine experts to grasp different views on the subject. The face-to-face discussion was supported by a document systematizing an initial agenda proposal and a brief presentation of the patient-centered care concept and theoretical elements that underpin its practice. Panel participants defined a set of items to be explored in studies to identify implementation and to strengthen and to measure strategies for patient-centered care in the Brazilian context.