NGT-3D: a simple nematode cultivation system to study Caenorhabditis elegans biology in 3D.
The nematodeCaenorhabditiselegansis one of the premier experimental model organisms today. In the laboratory, they display characteristic development, fertility, and behaviors in a two dimensional habitat. In nature, however,C. elegansis found in three dimensional environments such as rotting fruit. To investigate the biology ofC. elegansin a 3D controlled environment we designed a nematode cultivation habitat which we term the nematode growth tube or NGT-3D. NGT-3D allows for the growth of both nematodes and the bacteria they consume. Worms show comparable rates of growth, reproduction and lifespan when bacterial colonies in the 3D matrix are abundant. However, when bacteria are sparse, growth and brood size fail to reach levels observed in standard 2D plates. Using NGT-3D we observe drastic deficits in fertility in a sensory mutant in 3D compared to 2D, and this defect was likely due to an inability to locate bacteria. Overall, NGT-3D will sharpen our understanding of nematode biology and allow scientists to investigate questions of nematode ecology and evolutionary fitness in the laboratory.


Layered host-guest long-afterglow ultrathin nanosheets: high-efficiency phosphorescence energy transfer at 2D confined interface.
Tuning and optimizing the efficiency of light energy transfer play an important role in meeting modern challenges of minimizing energy loss and developing high-performance optoelectronic materials. However, attempts to fabricate systems giving highly efficient energy transfer between luminescent donor and acceptor have achieved limited success to date. Herein, we present a strategy towards phosphorescence energy transfer at a 2D orderly crystalline interface. We first show that new ultrathin nanosheet materials giving long-afterglow luminescence can be obtained by assembling aromatic guests into a layered double hydroxide host. Furthermore, we demonstrate that co-assembly of these long-lived energy donors with an energy acceptor in the same host generates an ordered arrangement of phosphorescent donor-acceptor pairs spatially confined within the 2D nanogallery, which affords energy transfer efficiency as high as 99.7%. Therefore, this work offers an alternative route to develop new types of long-afterglow nanohybrids and efficient light transfer systems with potential energy, illumination and sensor applications.


[Modulation of pyramidal responses of the sensomotor cortex after combined stimulation of the lateral hypothalamus and the sensomotor cortex of the rat].
Periodical pyramidal tract (PT) stimulation in freely moving rats has been shown to be accompanied by statistically significant increased functional activity of neuronal populations of the sensorimotor cortex (SMC) manifesting as potentiation of the primary positive phase of the pyramidal-cortical response (PCR). When periodical PT stimulation is combined with the lateral hypothalamus (LH) stimulation of the same periodicity, a statistically significant increase in the magnitude of potentiation of the PCR positive phase compared to that occurring without the LH activation is observed. When the periodical PT stimulation is combined with the LH and SMC stimulation of the same periodicity, the potentiation of the PSR positive phase increases significantly in comparison with the potentiation occurring without the SMC stimulation.


Epidermal growth factor receptor as a molecular determinant of glioblastoma response to dopamine receptor D2 inhibitors.
There are ongoing clinical trials exploring the efficacy of dopamine receptor D2 (DRD2) inhibition against glioblastomas, the most common primary brain tumor. Here we examine potential molecular determinants of this efficacy. The Cancer Genome Atlas glioblastoma database and other published mRNA profiles were used to analyze the DRD2 and epidermal growth factor receptor (EGFR) expression pattern. In vitro and in vivo responses to DRD2 inhibitors were determined using patient-derived xenograft (PDX) glioblastoma models. Immunohistochemical studies were performed on clinically annotated glioblastoma samples derived from patients treated with ONC201. Analysis of clinical glioblastoma specimens derived from independent patient cohorts revealed an inverse correlation between EGFR and DRD2 mRNA expression, with implication that signaling mediated by these proteins shares overlapping functions. In independent panels of PDX glioblastoma lines, high EGFR expression was associated with poor in vitro and in vivo response to DRD2 inhibitors, including haloperidol and ONC201. Moreover, ectopic expression of a constitutively active EGFR, variant (v)III, suppressed glioblastoma sensitivity to ONC201. DRD2 expression positively correlated with expression of rate-limiting enzymes for dopamine synthesis as well as dopamine secretion, suggesting contribution of autocrine DRD2 signaling. Analysis of specimens from patients treated with ONC201 (n = 15) showed an inverse correlation between the intensity of EGFR staining and clinical response. The median overall survival for patients with high and low EGFR staining was 162 and 373 days, respectively (0.037). High EGFR expression is a determinant of poor glioblastoma response to DRD2. This finding should inform future clinical trial designs.


Education for new role development: the Community Matron in England.
This paper is a report of an evaluation of the English national pilot education programme preparing Community Matrons to fulfil their role. Investment in community services has been important in introducing new ways of working in the United Kingdom National Health Service, particularly for patients with complex long-term conditions. Development of the Community Matron role in England is an exemplar in the creation of modern nursing careers that are fit for purpose. A mixed methods approach to data collection was adopted. This included documentary analysis of a range of sources used in the development and evaluation of the programme. In addition, during 2006, a self-administered questionnaire was sent to all Community Matrons undertaking the programme (n = 70), with a response rate of 67% (n = 47). Individual telephone interviews were conducted with 17 students and six mentors. A focus group was also undertaken with the education programme Development Team (n = 5). Quantitative data were analysed using SPSS and qualitative data analysed using content and thematic analyses. The majority of students (n = 25) believed that the programme had met their expectations and had helped them to achieve the functions of the Community Matron role as defined in national competence statements. However, 17 students experienced difficulties in the level of organisational readiness to support them in their role, including, for example, lack of facilitation of their work-based learning. In spite of the successful programme design, there is a need for organisations to develop their infrastructure to support new roles as well as offering protected time to learn in practice.


Physician Beliefs about Physical and Mental Competency of Patients Applying for Concealed Weapon Permits.
Law enforcement officials have asked health care providers to evaluate patient applications for concealed weapon permits. The current study was designed to examine physician beliefs regarding competency to carry a concealed weapon for patients with specific physical and mental conditions. Among 222 North Carolina physicians who participated in this survey (40% response rate), large variation and uncertainty existed for determining competency. Physicians most frequently chose mild dementia, post-traumatic stress disorder, and recent depression as conditions that would render a patient not competent to carry a concealed weapon. Male physicians and those owning a gun were more likely to deem a patient competent. Almost a third of physicians were unsure about competence for most conditions. Physicians asked to assess competency of patients to carry a concealed weapon have quite disparate views on competency and little confidence in their decisions. If physicians are expected to assess patient competence to carry a concealed weapon, more objective criteria and training are needed.


Glucosylated cholesterol in mammalian cells and tissues: formation and degradation by multiple cellular β-glucosidases.
The membrane lipid glucosylceramide (GlcCer) is continuously formed and degraded. Cells express two GlcCer-degrading β-glucosidases, glucocerebrosidase (GBA) and GBA2, located in and outside the lysosome, respectively. Here we demonstrate that through transglucosylation both GBA and GBA2 are able to catalyze in vitro the transfer of glucosyl-moieties from GlcCer to cholesterol, and vice versa. Furthermore, the natural occurrence of 1-O-cholesteryl-β-D-glucopyranoside (GlcChol) in mouse tissues and human plasma is demonstrated using LC-MS/MS and (13)C6-labeled GlcChol as internal standard. In cells, the inhibition of GBA increases GlcChol, whereas inhibition of GBA2 decreases glucosylated sterol. Similarly, in GBA2-deficient mice, GlcChol is reduced. Depletion of GlcCer by inhibition of GlcCer synthase decreases GlcChol in cells and likewise in plasma of inhibitor-treated Gaucher disease patients. In tissues of mice with Niemann-Pick type C disease, a condition characterized by intralysosomal accumulation of cholesterol, marked elevations in GlcChol occur as well. When lysosomal accumulation of cholesterol is induced in cultured cells, GlcChol is formed via lysosomal GBA. This illustrates that reversible transglucosylation reactions are highly dependent on local availability of suitable acceptors. In conclusion, mammalian tissues contain GlcChol formed by transglucosylation through β-glucosidases using GlcCer as donor. Our findings reveal a novel metabolic function for GlcCer.


Ignore growing patient interest in alternative medicine at your peril, MDs warned.
Canada now has an institute to study alternative medicine and seek evidence concerning it. The founder, endocrinologist Wah Jun Tze, says most physicians appreciate that the institute will seek to find evidence for unproven therapies. It recently named a research director, and expects to have protocols for randomized, controlled studies in place by the new year. Herbal remedies are one likely candidate for study.


Aquatic ecotoxicological indicators in life-cycle assessment.
This paper compares available options for the aquatic ecotoxicological effect factor component in life-cycle assessment (LCA). The effect factor is expressed here as the change in risk per unit change in cumulative exposure, delta effect/delta exposure. The comparison is restricted to approaches linked, implicitly as well as explicitly, to species-sensitivity distributions (SSDs). This draws on recent insights for chemical mixtures and identifies the implications of different model choices. In spite of the many options, assumptions, and areas for further research, it is concluded that a single effect factor basis represents the best available practice for use in LCA at this time, deltaPAF(ms)/deltaC = 0.5/HC50, where deltaPAF(ms) is the change in the (potentially affected) fraction (PAF) of species that experiences an increase in exposure above a specified effect level, accounting for the presence of complex background mixtures (ms), deltaC is the change in cumulative exposure concentration of the chemical of interest, and HC50 is the median, chronic hazardous concentration for regional, multiple-species systems. The resultant aquatic effect factors are risk-based and can be estimated readily for many chemicals using available methods, without the need to describe the entire SSDs and without the need for additional data. For example, the octanol-water partitioning coefficient provides a sufficient estimation basis for about 50% of existing chemicals that have a narcosis mode of action. This also is relevant in LCA for chemicals that are at low concentrations in the environment, concentrations below the biological thresholds at which more specific modes of action would be of relevance.


Effects of magnetic cell separation on monocyte adhesion to endothelial cells under flow.
Studies on monocyte adhesion are frequently limited by spontaneous changes of CD11b and CD62L during cell purification. Most isolation protocols for flow cytometric analysis that overcome this problem cannot be used when large numbers of living cells are needed for functional adhesion assays. This study investigated whether magnetic cell separation of monocytes with a paramagnetic bead against CD33 is a feasible method combining high yield with a low degree of spontaneous activation. As determined by flow cytometry, isolation of magnetically tagged monocytes at 4 degrees C did not alter the expression of CD11b and CD62L when compared to whole blood controls. Warming the cells slowly to room temperature immediately before starting the adhesion assay in a parallel plate flow chamber at 37 degrees C prevented further upregulation of adhesion molecules due to rewarming. When adhesion of magnetically tagged monocytes was compared with untouched monocytes that had been isolated via depletion of contaminating leukocytes, videomicroscopy showed that labelling CD33 neither affected rolling nor firm adhesion to human umbilical venous endothelial cells under flow. Finally, the subsequent upregulation of tissue factor expression on adherent monocytes indicates that magnetically separated monocytes responded properly to activating stimuli during cell adhesion. We conclude that magnetic cell separation via CD33 represents a feasible method for cell separation whenever large numbers of non-activated monocytes are needed for adhesion assays under flow.


The origin of cell surface immunoglobulin of marrow-derived and thymus-derived lymphocytes of the rat.
The origin of immunoglobulin on the surface of TDL in the rat has been studied by comparing the binding of purified alloantibodies recognizing the Ig-1a allotype of rat light chain, with that of rabbit antirat Fab antibodies. Both reagents labeled all TDL from rats of the DA strain (Ig-1a) with two categories of cells being detected; one binding 100-2,000 molecules of antibody, the other 10,000-100,000 molecules. These categories were likely to be synonomous with T and B cells, respectively. The [(125)I]antiallotype antibodies did not bind to TDL from rats of the PVG/c strain (Ig-1b). When the binding to TDL from (PVG/c x DA)F(1) animals was studied it was found that allelic exclusion occurred in the heavily labeled cells, but not in the lightly labeled ones. Furthermore, when lymphocytes of one allotype were transferred to irradiated recipients of the opposite allotype and recovered from the TDL or spleen of the recipient 20-30 h later, the immunoglobulin on heavily labeled cells was of the donor type, while that of lightly labeled ones bore the recipient marker. Thus heavily labeled cells (B lymphocytes) had synthesized their own immunoglobulin while lightly labeled cells (T lymphocytes) had acquired theirs passively by adsorption. The class of immunoglobulin on lightly labeled cells was also studied and it was found that [(125)I]anti-IgM antibodies bound to an extent approaching the [(125)I]anti-Fab binding, while [(125)I]anti-IgG(2a+2b) antibodies gave much less binding.


A multi-institutional survey on calvarial vault remodeling techniques for sagittal synostosis and outcomes analysis for patients treated at 12 months and older.
Surgical treatment of sagittal craniosynostosis is challenging in older patients. This study aimed to assess the effect of increasing age on open surgical technique selection and patient outcomes using the multi-institutional Synostosis Research Group (SynRG) collaboration. Surgeons in SynRG were surveyed for key influences on their preferred open calvarial vault remodeling techniques at various patient ages: < 6, 6-12, and > 12 months. The SynRG database was then queried for open repairs of nonsyndromic sagittal craniosynostosis performed for patients older than 12 months of age. Perioperative measures, complications, and preoperative and postoperative cephalic indices were reviewed. All surgeons preferred to treat patients at an earlier age, and most (89%) believed that less-optimal outcomes were achieved at ages older than 12 months. The modified pi procedure was the dominant technique in those younger than 12 months, while more involved open surgical techniques were performed for older patients, with a wide variety of open calvarial vault remodeling techniques used. Forty-four patients met inclusion criteria, with a mean (± SD) age at surgery of 29 ± 16 months. Eleven patients underwent parietal reshaping, 10 parietal-occipital switch, 9 clamshell craniotomy, 7 geometric parietal expansion, 6 modified pi procedure, and 1 parietal distraction. There were no readmissions, complications, or mortality within 30 days postoperatively. Patients' cephalic indices improved a mean of 6.4% ± 4.0%, with a mean postoperative cephalic index of 74.2% ± 4.9%. Differences in postoperative cephalic index (p < 0.04) and hospital length of stay (p = 0.01) were significant between technique cohorts. Post hoc Tukey-Kramer analysis identified the parietal reshaping technique as being significantly associated with a reduced hospital length of stay. Patient age is an important driver in technique selection, with surgeons selecting a more involved calvarial vault remodeling technique in older children. A variety of surgical techniques were analyzed, with the parietal reshaping technique being significantly associated with reduced length of stay; however, multiple perioperative factors may be contributory and require further analysis. When performed at high-volume centers by experienced pediatric neurosurgeons and craniofacial surgeons, open calvarial vault techniques can be a safe method for treating sagittal craniosynostosis in older children.


Visit-to-Visit Office Blood Pressure Variability and Cardiovascular Outcomes in SPRINT (Systolic Blood Pressure Intervention Trial).
Studies of visit-to-visit office blood pressure (BP) variability (OBPV) as a predictor of cardiovascular events and death in high-risk patients treated to lower BP targets are lacking. We conducted a post hoc analysis of SPRINT (Systolic Blood Pressure Intervention Trial), a well-characterized cohort of participants randomized to intensive (<120 mm Hg) or standard (<140 mm Hg) systolic BP targets. We defined OBPV as the coefficient of variation of the systolic BP using measurements taken during the 3-,6-, 9-, and 12-month study visits. In our cohort of 7879 participants, older age, female sex, black race, current smoking, chronic kidney disease, and coronary disease were independent determinants of higher OBPV. Use of thiazide-type diuretics or dihydropyridine calcium channel blockers was associated with lower OBPV whereas angiotensin-converting enzyme inhibitors or angiotensin receptor blocker use was associated with higher OBPV. There was no difference in OBPV in participants randomized to standard or intensive treatment groups. We found that OBPV had no significant associations with the composite end point of fatal and nonfatal cardiovascular events (n=324 primary end points; adjusted hazard ratio, 1.20; 95% confidence interval, 0.85-1.69, highest versus lowest quintile) nor with heart failure or stroke. The highest quintile of OBPV (versus lowest) was associated with all-cause mortality (adjusted hazard ratio, 1.92; confidence interval, 1.22-3.03) although the association of OBPV overall with all-cause mortality was marginal (P=0.07). Our results suggest that clinicians should continue to focus on office BP control rather than on OBPV unless definitive benefits of reducing OBPV are shown in prospective trials. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01206062.


Interspecific cell markers and lineage in mammals.
Study of cell lineage in the mammalian embryo has relied heavily on the use of chimeras to follow the fate of genetically marked cells in later development. Such studies have often been limited by the types of genetic markers available; there are very few markers that allow analysis of the spatial distribution of individual cells at all stages of development. We have developed a marker system that is based on the identification of cells of Mus musculus origin in M. musculus-M. caroli chimeras by in situ DNA-DNA hybridization using a cloned probe to M. musculus satellite DNA. This provides the first ubiquitous in situ cell marker system for mammalian chimeras. We have recently refined the system by the use of biotin-labelled probes and detection of hybridization by streptavidin-peroxidase binding. This increases both the speed and the resolution of the assay. We have used the marker for cell lineage analysis in both embryonic and adult chimeras and results from analysis of the derivatives of early cell lineages in later development and study of coherent growth versus cell mixing in the postimplantation embryo are presented. The importance of understanding embryonic cell lineages as a prelude to molecular studies is emphasized.


Management Principles and Current Debates Surrounding Common Pediatric Elbow Fractures.
Elbow fractures are among the most common fractures sustained in pediatric patients. A specific set of pediatric elbow fractures (olecranon, radial neck, and lateral condyle fractures) comprises the ones that occur most often. It is important to review commonly accepted principles in the evaluation and treatment of these injuries as well as highlight some debates that exist within the literature regarding the optimal management of these injuries. Although management of pediatric olecranon, radial neck, and lateral condyle fractures has been well described, controversy persists among orthopaedic surgeons regarding the surgical indications and preferred fixation techniques for these injuries.


A Bayesian spatial multimarker genetic random-effect model for fine-scale mapping.
Multiple markers in linkage disequilibrium (LD) are usually used to localize the disease gene location. These markers may contribute to the disease etiology simultaneously. In contrast to the single-locus tests, we propose a genetic random effects model that accounts for the dependence between loci via their spatial structures. In this model, the locus-specific random effects measure not only the genetic disease risk, but also the correlations between markers. In other words, the model incorporates this relation in both mean and covariance structures, and the variance components play important roles. We consider two different settings for the spatial relations. The first is our proposal, relative distance function (RDF), which is intuitive in the sense that markers nearby are likely to correlate with each other. The second setting is a common exponential decay function (EDF). Under each setting, the inference of the genetic parameters is fully Bayesian with Markov chain Monte Carlo (MCMC) sampling. We demonstrate the validity and the utility of the proposed approach with two real datasets and simulation studies. The analyses show that the proposed model with either one of two spatial correlations performs better as compared with the single locus analysis. In addition, under the RDF model, a more precise estimate for the disease locus can be obtained even when the candidate markers are fairly dense. In all simulations, the inference under the true model provides unbiased estimates of the genetic parameters, and the model with the spatial correlation structure does lead to greater confidence interval coverage probabilities.


Diffusion-weighted MRI findings of treated and untreated retroperitoneal fibrosis.
We aimed to evaluate diffusion-weighted imaging (DWI) findings in patients with treated and untreated retroperitoneal fibrosis (RPF). We analyzed magnetic resonance imaging examinations of 44 RPF patients (36 male, 8 female), of which 15 were untreated and 29 were under therapy. Qualitative DWI and T1 postcontrast signal intensities and the largest perivascular extent of RPF were compared between treated and untreated groups and correlated to erythrocyte sedimentation rate and C-reactive protein values. Quantitative DWI signal intensities and apparent-diffusion-coefficients were calculated in regions-of-interest, together with a relative index between signal intensities of RPF and psoas muscle in 15 untreated patients and 14 patients under treatment with remaining perivascular fibrosis of more than 5 mm. The extent of RPF in untreated patients was significantly larger compared with the extent of RPF in treated patients (P <0.0001). DWI signal intensities were significantly higher in untreated patients than in patients under therapy (mean, 27 s/mm2 vs. 20 s/mm2; P = 0.009). The calculated DWI-index was significantly higher in untreated patients than in patients under therapy (P = 0.003). Our data show significant differences in the DWI findings (b800 signal intensities and relative DWI-index) of patients with treated and untreated RPF. DWI is a promising technique in the assessment of disease activity and the selection of patients suitable for medical therapy.


Comparative proteomic analysis of extracellular proteins expressed by various clonal types of Staphylococcus aureus and during planktonic growth and biofilm development.
Staphylococcus aureus is well known for its biofilm formation with rapid emergence of new clones circulating worldwide. The main objectives of the study were (1) to identify possible differences in protein expression among various and closely related clonal types of S. aureus, (2) to establish the differences in protein expression in terms of size of protein spots and its intensities between bacteria which are grown statically (biofilm formation) with that of under aeration and agitation, and (3) to compare the differences in protein expression as a function of time (in hours). In this study, we selected six clinical isolates comprising two similar (MRSA-527 and MRSA-524) and four different (MRSA-139, MSSA-12E, MSSA-22d, and MSSA-10E) types identified by spa typing, MLST and SCCmec typing. We performed 2D gel migration comparison. Also, two MRSA isolates (527 and 139) were selected to determine quantitative changes in the level of extracellular proteins at different biofilm growth time points of 12, 24, and 48 h. The study was done using a strategy that combines 2-DGE and LC-MS/MS analysis for absolute quantification and identification of the extracellular proteins. The 2DGE revealed that the proteomic profiles for the isolates belonging to the similar spa, MLST, and SCCmec types were still quite different. Among the extracellular proteins secreted at different time points of biofilm formation, significant changes in protein expression were observed at 48 h incubation as compared to the exponential growth at 12 h incubation. The main conclusion of the work is that the authors do observe differences among isolates, and growth conditions do influence the protein content at different time points of biofilm formation.


Advances in first-principles modelling of point defects in UO2: f electron correlations and the issue of local energy minima.
Over the last decade, a significant amount of work has been devoted to point defect behaviour in UO2 using approximations beyond density functional theory (DFT), in particular DFT + U and hybrid functionals for correlated electrons. We review the results of these studies from calculations of bulk UO2 properties to the more recent determination of activation energies for self-diffusion in UO2, as well as a comparison with their experimental counterparts. We also discuss the efficiency of the three known methods developed to circumvent the presence of metastable states, namely occupation matrix control, U-ramping and quasi-annealing.


Symptoms of oropharyngeal dysphagia, efficiency, and safety of swallowing in patients after treatment for head and neck cancer.
The aim of this study is to investigate the relation between symptoms and signs of oropharyngeal dysphagia after treatment for head and neck cancer. An observational analytical study with retrospective and prospective components was carried out including 25 adult and elderly patients with head and neck cancer who had completed the treatment at least 3 months before data collection. Data from the Eating Assessment Tool (EAT-10) protocol were analyzed, as well as from the videofluoroscopic swallowing exams. Based on videofluoroscopy, the degree of oropharyngeal dysphagia was classified, as well as the safety and efficiency grade using the DIGEST (Dynamic Imaging Grade of Swallowing Toxicity) scale. Pearson's correlation test was applied, adopting a significance level of 5%. There was a significant correlation between question 4 of the EAT-10 (swallowing solids takes extra effort) and the efficiency profile (p = 0.004), as well as between question 4 and the DIGEST score (p = 0.002). No significant relation was found between the DIGEST score and EAT-10 total score (p = 0.180) and not even between EAT-10 total score and efficiency (p = 0.129) or safety grade (p = 0.878). In conclusion, no relation was found between most of the dysphagia signs and symptoms investigated, demonstrating that the individual's perception of the swallowing function may not be consistent with the findings of the instrumental evaluation after long-term treatment for head and neck cancer.


Use of adipose derived stem cells accelerates the healing process in third-degree burns.
Burns are defined as a traumatic injury, usually of thermal origin, that affects the epithelial and adjacent tissue and is classified according to the depth reached. Tissue repair involved in this type of injury is often a challenge both due to its severity and the multiplicity of complications. Regenerative medicine has focused on the use of low-level laser photobiomodulation therapy (LLLT) and adipose-derived stem cells (ADSC), especially in the early stages of the process, to promote better healing and shorten repair time. Therefore, aim of this study was to evaluate the action of LLLT (660 nm) and ADSC in the repair process of burned skin tissue and investigate the association of the techniques (LLLT and ADSC). An in vivo study was carried out using 96 rats (Wister) with a scald burn model at a temperature of 95ºC, exposing the animal's back for 14 s. Animals were randomized into seven groups and three periods, five, 14 and 21 days. The groups included GC: Control group, ADSC-: Group treated with CD49d negative cells, ADSC+ : Group treated with positive CD49d cells, CULT: Group treated with conventional isolation cells, LLLT: Group treated only with LLLT Low Power Laser, ADSC-LLLT: Group treated with CD49d negative cells and LLLT. ADSC+LLLT: Group treated with positive CD49d cells and LLLT. The groups treated with LLLT (660 nm; 5 J/cm2) received irradiation three times a week, on alternate days for five, 14 and 21 days, according to the time of biopsy. ADSC-treated groups received one to three applications of the cells in a total volume of 1000 μL starting soon after the surgical debridement of the burn. Photographic monitoring was carried out at 5, 14 and 21 days after the beginning of the experiment to assess the degree of lesion contraction. Macroscopic, morphometric and histopathological analyzes were performed. We showed significant re-epithelialization as well as an improvement in the healing process in the ADSC+, LLLT and ADSC+LLLT groups. We observed effects in the reduction of the inflammatory phase, increase in angiogenesis, decrease in oedema, greater collagen deposition, and better organization of the extracellular matrix compared to the other treatments. Moreover, the immunomagnetic separation of ADSC cells through the expression of the CD49d protein proved to be a useful means to obtain a more homogeneous population of cells with a role in tissue regeneration compared to the ADSC- and CULT groups. In conclusion, the association of ADSC+ with LLLT was effective in accelerating the burn repair process, stimulating cell proliferation and formation of more normal skin tissue.


Genetic markers at the leptin (OB) locus are not significantly linked to hypertension in African Americans.
Increased body mass index (BMI) has been correlated with increased blood pressure in human populations. To examine the role of the leptin gene (OB) in essential hypertension in African Americans, we performed affected sib pair analysis on a set of 103 hypertensive African American sibships using four highly polymorphic markers at the human leptin locus. No evidence of linkage was detected between these markers and the phenotype of essential hypertension either in these sibships or in a severely obese subset of 46 sibships in which each sibling had a BMI > or = 85th percentile for the US population. Using BMI rather than hypertension as a quantitative trait, we found significant linkage for the marker D7S504 (P=0.029) but not for the other markers. Significance strengthened in the overweight subset of sibships for this marker (P=0.001), and there was a trend of lower P values for the other three markers. However, multipoint analysis with the use of all four markers simultaneously to estimate linkage between BMI and the leptin locus did not demonstrate a statistically significant relationship. Analysis of the coding region of the leptin gene (exons 2 and 3) by single-strand conformational polymorphism revealed a rare Ile-Val polymorphism at amino acid 45 but revealed no other alterations. These results suggest that the OB gene is not a major contributor to the phenotype of essential hypertension in African Americans, although a minor contribution to the phenotype of extreme obesity in this group cannot be ruled out.


Swabbing students: should universities be allowed to facilitate educational DNA testing?
Recognizing the profound need for greater patient and provider familiarity with personalized genomic medicine, many university instructors are including personalized genotyping as part of their curricula. During seminars and lectures students run polymerase chain reactions on their own DNA or evaluate their experiences using direct-to-consumer genetic testing services subsidized by the university. By testing for genes that may influence behavioral or health-related traits, however, such as alcohol tolerance and cancer susceptibility, certain universities have stirred debate on the ethical concerns raised by educational genotyping. Considering the potential for psychosocial harm and medically relevant outcomes, how far should university-facilitated DNA testing be permitted to go? The analysis here distinguishes among these learning initiatives and critiques their approaches to the ethical concerns raised by educational genotyping.


Eif4enif1 haploinsufficiency disrupts oocyte mitochondrial dynamics and leads to subfertility.
Infertility affects couples worldwide. Premature ovarian insufficiency (POI) refers to loss of ovarian function before 40 years of age and is a contributing factor to infertility. Several case studies have reported dominant-inherited POI symptoms in families with heterozygous EIF4ENIF1 (4E-T) mutations. However, the effects of EIF4ENIF1 haploinsufficiency have rarely been studied in animal models to reveal the underlying molecular changes related to infertility. Here, we demonstrate that Eif4enif1 haploinsufficiency causes mouse subfertility, impairs oocyte maturation and partially arrests early embryonic development. Using dual-omic sequencing, we observed that Eif4enif1 haploinsufficiency significantly altered both transcriptome and translatome in mouse oocytes, by which we further revealed oocyte mitochondrial hyperfusion and mitochondria-associated ribonucleoprotein domain distribution alteration in Eif4enif1-deficient oocytes. This study provides new insights into the molecular mechanisms underlying clinical fertility failure and new avenues to pursue new therapeutic targets to address infertility.


Mode of selection and experimental evolution of antifungal drug resistance in Saccharomyces cerevisiae.
We show that mode of selection, degree of dominance of mutations, and ploidy are determining factors in the evolution of resistance to the antifungal drug fluconazole in yeast. In experiment 1, yeast populations were subjected to a stepwise increase in fluconazole concentration over 400 generations. Under this regimen, two mutations in the same two chromosomal regions rose to high frequency in parallel in three replicate populations. These mutations were semidominant and additive in their effect on resistance. The first of these mutations mapped to PDR1 and resulted in the overexpression of the ABC transporter genes PDR5 and SNQ2. These mutations had an unexpected pleiotropic effect of reducing the residual ability of the wild type to reproduce at the highest concentrations of fluconazole. In experiment 2, yeast populations were subjected to a single high concentration of fluconazole. Under this regimen, a single recessive mutation appeared in each of three replicate populations. In a genome-wide screen of approximately 4700 viable deletion strains, 13 were classified as resistant to fluconazole (ERG3, ERG6, YMR102C, YMR099C, YPL056C, ERG28, OSH1, SCS2, CKA2, SML1, YBR147W, YGR283C, and YLR407W). The mutations in experiment 2 all mapped to ERG3 and resulted in the overexpression of the gene encoding the drug target ERG11, but not PDR5 and SNQ2. Diploid hybrids from experiments 1 and 2 were less fit than the parents in the presence of fluconazole. In a variation of experiment 2, haploids showed a higher frequency of resistance than diploids, suggesting that degree of dominance and ploidy are important factors in the evolution of antifungal drug resistance.


An AFLP-based genome-wide mapping strategy.
To efficiently determine the chromosomal location of phenotypic mutants, we designed a genome-wide mapping strategy that can be used in any crop for which a dense AFLP (Amplified Fragment Length Polymorphism) map is available or can be made. The AFLP technique is particularly suitable to initiate map-based cloning projects because it detects many markers per reaction. First a standard set of AFLP primer combinations that results in a framework of AFLP markers well dispersed over the genome is selected. These primer combinations are applied to a limited number of mutant individuals from a segregating population to register linkage and non-linkage of the AFLP markers to the gene-of-interest. Further delineation of the area of interest is accomplished by analyzing the remaining recombinants and additional mutant individuals with AFLP markers that lie within the identified region. We illustrate the usefulness of the method by mapping three rotunda ( ron) leaf-form mutant loci of Arabidopsis thaliana and show that in the initial phase of map-based cloning projects a 400-600 kb interval can be identified for the average mutant locus within a few weeks. Once such an area is identified and before initiating the more time-consuming fine-mapping procedure, it is essential to examine publicly available databases for candidate genes and known mutants in the identified region. The 390-kb interval on chromosome 4 that harbors the ron2 mutation, also carries a known flower mutant, leunig ( lug); upon crossing, the two mutants appeared to be allelic. When no such candidates are found, the mapping procedure should be continued. We present a strategy to efficiently select recombinants that can be used for fine mapping.


Novel dipeptidyl peptidase IV resistant analogues of glucagon-like peptide-1(7-36)amide have preserved biological activities in vitro conferring improved glucose-lowering action in vivo.
Although the incretin hormone glucagon-like peptide-1 (GLP-1) is a potent stimulator of insulin release, its rapid degradation in vivo by the enzyme dipeptidyl peptidase IV (DPP IV) greatly limits its potential for treatment of type 2 diabetes. Here, we report two novel Ala(8)-substituted analogues of GLP-1, (Abu(8))GLP-1 and (Val(8))GLP-1 which were completely resistant to inactivation by DPP IV or human plasma. (Abu(8))GLP-1 and (Val(8))GLP-1 exhibited moderate affinities (IC(50): 4.76 and 81.1 nM, respectively) for the human GLP-1 receptor compared with native GLP-1 (IC(50): 0.37 nM). (Abu(8))GLP-1 and (Val(8))GLP-1 dose-dependently stimulated cAMP in insulin-secreting BRIN BD11 cells with reduced potency compared with native GLP-1 (1.5- and 3.5-fold, respectively). Consistent with other mechanisms of action, the analogues showed similar, or in the case of (Val(8))GLP-1 slightly impaired insulin releasing activity in BRIN BD11 cells. Using adult obese (ob/ob) mice, (Abu(8))GLP-1 had similar glucose-lowering potency to native GLP-1 whereas the action of (Val(8))GLP-1 was enhanced by 37%. The in vivo insulin-releasing activities were similar. These data indicate that substitution of Ala(8) in GLP-1 with Abu or Val confers resistance to DPP IV inactivation and that (Val(8))GLP-1 is a particularly potent N-terminally modified GLP-1 analogue of possible use in type 2 diabetes.


Generation by a widely applicable approach of a hybrid dioxygenase showing improved oxidation of polychlorobiphenyls.
Recently, a sequence-based approach has been developed for the fast isolation and characterization of class II aryl-hydroxylating dioxygenase activities (S. Kahl and B. Hofer, Microbiology 149:1475-1481, 2003). It comprises the PCR amplification of segments of alpha subunit genes of unknown sequence that encode the catalytic center and their fusion with sequences of the bphA gene cluster of Burkholderia xenovorans LB400. One of the resulting chimeric enzymes, harboring the core segment of a dioxygenase from Pseudomonas sp. strain B4-Magdeburg, has now been characterized with respect to the oxidation of chlorobiphenyls (CBs). Its substrate and product specificities differed favorably from those of the parental dioxygenase of strain LB400. The hybrid possessed a higher regiospecificity and yielded less unproductive dioxygenations at meta and para carbons. It attacked ortho-, meta-, and para-chlorinated rings with comparable efficiencies. It gave significantly higher yields in ortho,meta-dioxygenation of recalcitrant congeners containing a doubly ortho-chlorinated ring. While the parental enzyme yielded mainly unproductive meta, para dioxygenation of 2,5,4'-CB, the hybrid predominantly converted this congener into an ortho,meta-dioxygenated product. The subsequent enzymes of the LB400 catabolic pathway were able to transform most of the metabolites formed by the novel dioxygenase, indicating that the substrate ranges of these biocatalysts are not adapted to that of their initial pathway enzyme. Some of the catabolites, however, were identified as problematic for further degradation. Our results demonstrate that the outlined approach can successfully be applied to obtain novel dioxygenase specificities that favorably complement or supplement known ones.


Generalizing movement patterns following shoulder fixation.
Generalizing newly learned movement patterns beyond the training context is challenging for most motor learning situations. Here we tested whether learning of a new physical property of the arm during self-initiated reaching generalizes to new arm configurations. Human participants performed a single-joint elbow reaching task and/or countered mechanical perturbations that created pure elbow motion with the shoulder joint free to rotate or locked by the manipulandum. With the shoulder free, we found activation of shoulder extensor muscles for pure elbow extension trials, appropriate for countering torques that arise at the shoulder due to forearm rotation. After locking the shoulder joint, we found a partial reduction in shoulder muscle activity, appropriate because locking the shoulder joint cancels the torques that arise at the shoulder due to forearm rotation. In our first three experiments, we tested whether and to what extent this partial reduction in shoulder muscle activity generalizes when reaching in different situations: 1) different initial shoulder orientation, 2) different initial elbow orientation, and 3) different reach distance/speed. We found generalization for the different shoulder orientation and reach distance/speed as measured by a reliable reduction in shoulder activity in these situations but no generalization for the different elbow orientation. In our fourth experiment, we found that generalization is also transferred to feedback control by applying mechanical perturbations and observing reflex responses in a distinct shoulder orientation. These results indicate that partial learning of new intersegmental dynamics is not sufficient for modifying a general internal model of arm dynamics.NEW & NOTEWORTHY Here we show that partially learning to reduce shoulder muscle activity following shoulder fixation generalizes to other movement conditions, but it does not generalize globally. These findings suggest that the partial learning of new intersegmental dynamics is not sufficient for modifying a general internal model of the arm's dynamics.


Changes in EMG power spectrum (high-to-low ratio) with force fatigue in humans.
During and following high-load fatiguing voluntary contractions, the force response of skeletal muscle to electrical stimulation is altered so that the frequency-force curve is moved to the right. Fatiguing contractions also result in a shift to the left of the electromyographic (EMG) power spectrum. In the quadriceps muscle and the diaphragm of normal subjects the change in the force response to electrical stimulation has been correlated with the EMG changes. After repeated submaximal contractions in the quadriceps and diaphragm, the forces produced by electrical stimulation at low frequencies were reduced, indicating low-frequency fatigue. This type of fatigue persisted for several hours but did not result in any change in the EMG high-to-low ratio. Low-frequency fatigue is probably an important aspect of the failure of skeletal muscle to generate adequate force, and the EMG high-to-low ratio may not recognize this type of fatigue.


The chemistry behind antioxidant capacity assays.
This review summarizes the multifaceted aspects of antioxidants and the basic kinetic models of inhibited autoxidation and analyzes the chemical principles of antioxidant capacity assays. Depending upon the reactions involved, these assays can roughly be classified into two types: assays based on hydrogen atom transfer (HAT) reactions and assays based on electron transfer (ET). The majority of HAT-based assays apply a competitive reaction scheme, in which antioxidant and substrate compete for thermally generated peroxyl radicals through the decomposition of azo compounds. These assays include inhibition of induced low-density lipoprotein autoxidation, oxygen radical absorbance capacity (ORAC), total radical trapping antioxidant parameter (TRAP), and crocin bleaching assays. ET-based assays measure the capacity of an antioxidant in the reduction of an oxidant, which changes color when reduced. The degree of color change is correlated with the sample's antioxidant concentrations. ET-based assays include the total phenols assay by Folin-Ciocalteu reagent (FCR), Trolox equivalence antioxidant capacity (TEAC), ferric ion reducing antioxidant power (FRAP), "total antioxidant potential" assay using a Cu(II) complex as an oxidant, and DPPH. In addition, other assays intended to measure a sample's scavenging capacity of biologically relevant oxidants such as singlet oxygen, superoxide anion, peroxynitrite, and hydroxyl radical are also summarized. On the basis of this analysis, it is suggested that the total phenols assay by FCR be used to quantify an antioxidant's reducing capacity and the ORAC assay to quantify peroxyl radical scavenging capacity. To comprehensively study different aspects of antioxidants, validated and specific assays are needed in addition to these two commonly accepted assays.


The delayed rod afterimage.
A flashed background, presented to a dark-adapted eye, can saturate the rod system, making an incremental test patch invisible. But as the afterimage decays, the test can be distinguished. Increment thresholds measured within the decaying afterimage exhibit Weber's law over a wide range. The Penn and Hagins model of rod kinetics correctly predicts Weber's law, but makes incorrect predictions of the latency for the detection to occur. A new model, involving two exponential decays, is able to accommodate the latency data, as well as Weber's law. The model also makes good predictions of the results when the stimulus duration is increased from 100 msec to 1 sec.


Survival case of Vibrio vulnificus infection with multiple endocrine neoplasia type I.
A 35-year-old female with malabsorption syndrome who underwent a pancreatoduodenectomy for multiple endocrine adenomatosis 13 years prior was admitted to our hospital with diarrhea, general fatigue, high fever, and eruption in the lower legs. The patient had consumed raw shrimp a few days before onset and presented systemic inflammatory response syndrome at the time of hospitalization. Vibrio vulnificus was isolated from a blood culture performed before admission to the intensive care unit. We excised necrotizing tissue in the legs after improvement of her general condition. During the treatment process, glucose, catecholamine, and appropriate antibiotics were administered for hypoglycemia, hypotension, and high fever, respectively. The patient was discharged 107 days after contracting the disease. Of 18 septic patients with V. vulnificus infection admitted to our hospital, this was the first to develop septicemia in the absence of a previous liver dysfunction. In order to prevent this type of fatal infection, public education for immuno-compromised individuals as well as those with liver disease is essential. For early diagnosis and appropriate treatment, more effective strategies are required, such as the establishment of a network system where family physicians and emergency hospital staff could discuss information regarding high-risk patients.


D3 Receptors Regulate Excitability in a Unique Class of Prefrontal Pyramidal Cells.
The D3 dopamine receptor, a member of the Gi-coupled D2 family of dopamine receptors, is expressed throughout limbic circuits affected in neuropsychiatric disorders, including prefrontal cortex (PFC). These receptors are important for prefrontal executive function because pharmacological and genetic manipulations that affect prefrontal D3 receptors alter anxiety, social interaction, and reversal learning. However, the mechanisms by which D3 receptors regulate prefrontal circuits and whether D3 receptors regulate specific prefrontal subnetworks remains unknown. Here, we combine dopamine receptor reporter lines, anatomical tracing techniques, and electrophysiology to show that D3 receptor expression defines a novel subclass of layer 5 glutamatergic pyramidal cell in mouse PFC (either sex). D3-receptor-expressing pyramidal neurons are electrophysiologically and anatomically separable from neighboring neurons expressing D1 or D2 receptors based on their dendritic morphology and subthreshold and suprathreshold intrinsic excitability. D3-receptor-expressing neurons send axonal projections to intratelencephalic (IT) targets, including contralateral cortex, nucleus accumbens, and basolateral amygdala. Within these neurons, D3 receptor activation was found to regulate low-voltage-activated CaV3.2 calcium channels localized to the axon initial segment, which suppressed action potential (AP) excitability, particularly when APs occurred at high frequency. Therefore, these data indicate that D3 receptors regulate the excitability of a unique, IT prefrontal cell population, thereby defining novel circuitry and cellular actions for D3 receptors in PFC.SIGNIFICANCE STATEMENT The D3 dopamine receptor, a member of the Gi-coupled D2 family of dopamine receptors, are expressed throughout limbic circuits, including prefrontal cortex (PFC). They are of broad interest as a site for therapeutic intervention in serious mental illness, yet we know very little about their distribution or function within PFC. Here, we show that D3 receptors define a unique population of glutamatergic principal cells in mouse PFC that largely lack expression of D1 or D2 receptors. Within these cells, we find that D3 receptors regulate the ability to generate high-frequency action potential bursts through mechanisms not supported by other dopamine receptors. These results define unique circuitry and cellular actions for D3 receptors in regulating PFC networks.


Preparing facilitators from community-based organizations for evidence-based intervention training in Second Life.
A major barrier to the use and scale-up of evidence-based interventions are challenges related to training and capacity building. A cost-effective and highly interactive multi-user virtual environment, Second Life (SL) is a promising alternative for comprehensive face-to-face facilitator training. The purpose of this study was to examine the feasibility of using SL to train facilitators from community-based organizations to use ¡Cuídate! (Take Care of Yourself), one of the few evidence-based interventions developed and tested with Latino youth to reduce sexual risk behaviors. We recruited 35 participants from community-based organizations throughout the United States to participate in the SL ¡Cuídate! Training of Facilitators. Preparation to use SL consisted of four phases: (1) recruitment and computer capacity screening, (2) enrollment, (3) orientation to the SL program, and (4) technical support throughout the synchronous training sessions. Technical difficulties, the associated cause, and the mitigation strategy implemented were recorded during each session. Participants completed evaluations including perceptions of self-efficacy and confidence to complete the necessary skills to participate in SL training. Overall, participants reported high levels of self-efficacy for all skills necessary to participate in SL training. Based on an 11-point scale (0-10), self-efficacy to download and access the software was rated the highest: mean 8.29 (SD 2.19). Interacting with items in SL had the lowest mean score: mean 7.49 (SD 2.89). The majority of technical difficulties experienced by participants were related to inadequate Internet connections or computer malfunctions. Our findings support the feasibility of using SL for the ¡Cuídate! Training of Facilitators. The process used in this study to prepare participants to use SL can be used as a basis for other evidence-based intervention training in SL. This study is an important contribution to developing cost-effective and accessible training options for evidence-based interventions.


Hepatocyte turnover and regeneration: virtually a virtuoso performance.
The liver and exocrine pancreas share a common structure, with functioning units (hepatic plates and pancreatic acini) connected to the ductal tree. Here we show that Sox9 is expressed throughout the biliary and pancreatic ductal epithelia, which are connected to the intestinal stem-cell zone. Cre-based lineage tracing showed that adult intestinal cells, hepatocytes and pancreatic acinar cells are supplied physiologically from Sox9-expressing progenitors. Combination of lineage analysis and hepatic injury experiments showed involvement of Sox9-positive precursors in liver regeneration. Embryonic pancreatic Sox9-expressing cells differentiate into all types of mature cells, but their capacity for endocrine differentiation diminishes shortly after birth, when endocrine cells detach from the epithelial lining of the ducts and form the islets of Langerhans. We observed a developmental switch in the hepatic progenitor cell type from Sox9-negative to Sox9-positive progenitors as the biliary tree develops. These results suggest interdependence between the structure and homeostasis of endodermal organs, with Sox9 expression being linked to progenitor status.


Molecular and metabolic basis for the metabolic disorder normotriglyceridemic abetalipoproteinemia.
We have previously described a disorder, normotriglyceridemic abetalipoproteinemia, that is characterized by the virtual absence of plasma low density lipoproteins and complete absence of apoB-100, but with apparently normal secretion of triglyceride-rich lipoproteins containing apoB-48. The patient's plasma lipoproteins were shown on polyacrylamide gels and by antibody mapping to have a new truncated apoB variant, apoB-50, circulating along with her apoB-48. We have found this individual to be homozygous for a single C-to-T nucleotide substitution at apoB codon 2252, which produces a premature in-frame stop codon. Thus, this is a rare example of homozygous hypobetalipoproteinemia. Electron photomicrographs revealed that the diameters of particles in the d less than 1.006 g/ml lipoprotein fraction, in both the postprandial and postabsorptive state, are bimodally distributed. The molar ratio of apoE to apoB in these particles is 3.5:1, similar to normal VLDL. The plasma LDL interval contains both spherical and cuboidal particles. Autologous reinfusion of labeled d less than 1.006 g/ml lipoproteins showed exponential disappearance from plasma, with an apparent half-removal time of 50 min, somewhat slower than for normal chylomicrons but within the normal range for VLDL. The calculated production rate for apoB was within the normal range in this subject. A very small amount of label was found briefly in the IDL fraction, but none at any time in LDL or HDL. Therefore, because LDL particles that contain apoB-50 lack the putative ligand domain of the LDL receptor, we conclude that the very low level of LDL is due to the rapid removal of the abnormal VLDL particles before their conversion to LDL can take place.


A kinetic model of the evolution of a protein interaction network.
Known protein interaction networks have very particular properties. Old proteins tend to have more interactions than new ones. One of the best statistical representatives of this property is the node degree distribution (distribution of proteins having a given number of interactions). It has previously been shown that this distribution is very close to the sum of two distinct exponential components. In this paper, we asked: What are the possible mechanisms of evolution for such types of networks? To answer this question, we tested a kinetic model for simplified evolution of a protein interactome. Our proposed model considers the emergence of new genes and interactions and the loss of old ones. We assumed that there are generally two coexisting classes of proteins. Proteins constituting the first class are essential only for ecological adaptations and are easily lost when ecological conditions change. Proteins of the second class are essential for basic life processes and, hence, are always effectively protected against deletion. All proteins can transit between the above classes in both directions. We also assumed that the phenomenon of gene duplication is always related to ecological adaptation and that a new copy of a duplicated gene is not essential. According to this model, all proteins gain new interactions with a rate that preferentially increases with the number of interactions (the rich get richer). Proteins can also gain interactions because of duplication. Proteins lose their interactions both with and without the loss of partner genes. The proposed model reproduces the main properties of protein-protein interaction networks very well. The connectivity of the oldest part of the interaction network is densest, and the node degree distribution follows the sum of two shifted power-law functions, which is a theoretical generalization of the previous finding. The above distribution covers the wide range of values of node degrees very well, much better than a power law or generalized power law supplemented with an exponential cut-off. The presented model also relates the total number of interactome links to the total number of interacting proteins. The theoretical results were for the interactomes of A. thaliana, B. taurus, C. elegans, D. melanogaster, E. coli, H. pylori, H. sapiens, M. musculus, R. norvegicus and S. cerevisiae. Using these approaches, the kinetic parameters could be estimated. Finally, the model revealed the evolutionary kinetics of proteome formation, the phenomenon of protein differentiation and the process of gaining new interactions.


Amplification of the c-myc oncogene in non-small cell lung cancer.
Fresh non-small cell lung carcinoma surgical specimens were taken from 17 patients and 3 controls and screened for genetic abnormalities of the c-myc oncogene. Southern blot hybridization analysis demonstrated two- to fivefold amplification of the c-myc gene in 10 cases, i.e., 7 of 13 epidermoid lung carcinomas, 2 of 3 adenocarcinomas and 1 of 1 osteogenic sarcoma metastatic to the lung. Two- to fivefold amplification was observed in tissues from stage III and IV epidermoid carcinomas and adenocarcinomas of the lung. A correlation between cancer stage and c-myc gene amplification was found.


Terminal axon branching is regulated by the LKB1-NUAK1 kinase pathway via presynaptic mitochondrial capture.
The molecular mechanisms underlying the axon arborization of mammalian neurons are poorly understood but are critical for the establishment of functional neural circuits. We identified a pathway defined by two kinases, LKB1 and NUAK1, required for cortical axon branching in vivo. Conditional deletion of LKB1 after axon specification or knockdown of NUAK1 drastically reduced axon branching in vivo, whereas their overexpression was sufficient to increase axon branching. The LKB1-NUAK1 pathway controls mitochondria immobilization in axons. Using manipulation of Syntaphilin, a protein necessary and sufficient to arrest mitochondrial transport specifically in the axon, we demonstrate that the LKB1-NUAK1 kinase pathway regulates axon branching by promoting mitochondria immobilization. Finally, we show that LKB1 and NUAK1 are necessary and sufficient to immobilize mitochondria specifically at nascent presynaptic sites. Our results unravel a link between presynaptic mitochondrial capture and axon branching.


Motor speech signature of behavioral variant frontotemporal dementia: Refining the phenotype.
To provide a comprehensive description of motor speech function in behavioral variant frontotemporal dementia (bvFTD). Forty-eight individuals (24 bvFTD and 24 age- and sex-matched healthy controls) provided speech samples. These varied in complexity and thus cognitive demand. Their language was assessed using the Progressive Aphasia Language Scale and verbal fluency tasks. Speech was analyzed perceptually to describe the nature of deficits and acoustically to quantify differences between patients with bvFTD and healthy controls. Cortical thickness and subcortical volume derived from MRI scans were correlated with speech outcomes in patients with bvFTD. Speech of affected individuals was significantly different from that of healthy controls. The speech signature of patients with bvFTD is characterized by a reduced rate (75%) and accuracy (65%) on alternating syllable production tasks, and prosodic deficits including reduced speech rate (45%), prolonged intervals (54%), and use of short phrases (41%). Groups differed on acoustic measures derived from the reading, unprepared monologue, and diadochokinetic tasks but not the days of the week or sustained vowel tasks. Variability of silence length was associated with cortical thickness of the inferior frontal gyrus and insula and speech rate with the precentral gyrus. One in 8 patients presented with moderate speech timing deficits with a further two-thirds rated as mild or subclinical. Subtle but measurable deficits in prosody are common in bvFTD and should be considered during disease management. Language function correlated with speech timing measures derived from the unprepared monologue only.


Cell-Membrane-Derived Nanoparticles with Notch-1 Suppressor Delivery Promote Hypoxic Cell-Cell Packing and Inhibit Angiogenesis Acting as a Two-Edged Sword.
Cell-cell interactions regulate intracellular signaling via reciprocal contacts of cell membranes in tissue regeneration and cancer growth, indicating a critical need of membrane-derived tools in studying these processes. Hence, cell-membrane-derived nanoparticles (CMNPs) are produced using tonsil-derived mesenchymal stem cells (TMSCs) from children owing to their short doubling time. As target cell types, laryngeal cancer cells are compared to bone-marrow-derived MSCs (BMSCs) because of their cartilage damaging and chondrogenic characteristics, respectively. Treating spheroids of these cell types with CMNPs exacerbates interspheroid hypoxia with robust maintenance of the cell-cell interaction signature for 7 days. Both cell types prefer a hypoxic environment, as opposed to blood vessel formation that is absent in cartilage but is required for cancer growth. Hence, angiogenesis is inhibited by displaying the Notch-1 aptamer on CMNPs. Consequently, laryngeal cancer growth is suppressed efficiently in contrast to improved chondroprotection observed in a series of cell and animal experiments using a xenograft mouse model of laryngeal cancer. Altogether, CMNPs execute a two-edged sword function of inducing hypoxic cell-cell packing, followed by suppressing angiogenesis to promote laryngeal cancer death and chondrogenesis simultaneously. This study presents a previously unexplored therapeutic strategy for anti-cancer and chondroprotective treatment using CMNPs.


Optical waveguides formed by thermal migration of ions in glass.
We present our results on optical waveguides formed by thermal diffusion of ions in glass. It was found that the peak of the ion-exchanged region can be shifted into the substrate interior by limiting the diffusion process. We also found that low loss films (<0.1 dB/cm) can be fabricated using this process and that the modal losses in these films do not agree with those losses predicted by existing theories. Also, the ion-exchange process has proved to be a simple means for fabricating tapered-edge couplers.


Frequency entrainment in long chains of oscillators with random natural frequencies in the weak coupling limit.
We study oscillator chains of the form phi; (k) = omega(k) +K[Gamma( phi(k-1) - phi(k) )+Gamma( phi(k+1) - phi(k) )], where phi(k) epsilon[0,2pi) is the phase of oscillator k. In the thermodynamic limit where the number of oscillators goes to infinity, for suitable choices of Gamma(x), we prove that there is a critical coupling strength K(c), above which a stable frequency-entrained state exists, but below which the probability is zero to have such a state. It is assumed that the natural frequencies are random with finite bandwidth. A crucial condition on Gamma(x) is that it is nonodd, i.e., mid R: Gamma(x)+Gamma(-x) mid R: not equal 0. The interest in the results comes from the fact that any chain of limit-cycle oscillators can be described by equations of the above form in the limits of weak coupling and narrow distribution of natural frequencies.


Susceptibility to chronic pain following nerve injury is genetically affected by CACNG2.
Chronic neuropathic pain is affected by specifics of the precipitating neural pathology, psychosocial factors, and by genetic predisposition. Little is known about the identity of predisposing genes. Using an integrative approach, we discovered that CACNG2 significantly affects susceptibility to chronic pain following nerve injury. CACNG2 encodes for stargazin, a protein intimately involved in the trafficking of glutamatergic AMPA receptors. The protein might also be a Ca(2+) channel subunit. CACNG2 has previously been implicated in epilepsy. Initially, using two fine-mapping strategies in a mouse model (recombinant progeny testing [RPT] and recombinant inbred segregation test [RIST]), we mapped a pain-related quantitative trait locus (QTL) (Pain1) into a 4.2-Mb interval on chromosome 15. This interval includes 155 genes. Subsequently, bioinformatics and whole-genome microarray expression analysis were used to narrow the list of candidates and ultimately to pinpoint Cacng2 as a likely candidate. Analysis of stargazer mice, a Cacng2 hypomorphic mutant, provided electrophysiological and behavioral evidence for the gene's functional role in pain processing. Finally, we showed that human CACNG2 polymorphisms are associated with chronic pain in a cohort of cancer patients who underwent breast surgery. Our findings provide novel information on the genetic basis of neuropathic pain and new insights into pain physiology that may ultimately enable better treatments.


Medicare patient access to posthospital skilled nursing facility care.
The willingness of nursing homes to accept any Medicare admissions (Medicare participation) and the number of patients they serve (Medicare utilization) affect the access of Medicare SNF patients to posthospital care. Characteristics of facilities, their market areas, and state Medicaid reimbursement were found to affect both participation and utilization. Most important, when Medicaid and private nursing home market conditions support high-intensity care, facilities are more likely to serve Medicare patients and admit more of them. SNFs in states that require Medicare certification are more likely to accept Medicare admissions, suggesting that the cost and effort of achieving certification are a barrier to service to Medicare SNF patients. For-profit facilities, hospital-based SNFs, and larger nursing homes have higher rates of participation and serve more Medicare patients when they serve any.


CHFR promoter methylation indicates poor prognosis in stage II microsatellite stable colorectal cancer.
Data on the prognostic significance of promoter CpG island methylation in colorectal cancer (CRC) are conflicting, possibly due to associations between methylation and other factors affecting survival such as genetic alterations and use of adjuvant therapy. Here, we examine the prognostic impact of promoter methylation in patients with CRC treated with surgery alone in the context of microsatellite instability (MSI), BRAF and KRAS mutations. One hundred and seventy-three CRCs were analyzed for promoter methylation of 19 tumor suppressor and DNA repair genes, the CpG island methylator phenotype (CIMP), MSI, the exon 15 V600E BRAF mutation and KRAS codon 12 and 13 mutations. Unsupervised hierarchical clustering based on methylation status of 19 genes revealed three subgroups: cluster 1 [CL1, 57% (98/173) of CRCs], cluster 2 [CL2, 25% (43/173) of CRCs], and cluster 3 [CL3, 18% (32/173) of CRCs]. CL3 had the highest methylation index (0.25, 0.49, and 0.69, respectively, P = <0.01) and was strongly associated with CIMP (P < 0.01). Subgroup analysis for tumor stage, MSI, and BRAF status showed no statistically significant differences in survival between CL1, CL2, and CL3 nor between CIMP and non-CIMP CRCs. Analyzing genes separately revealed that CHFR promoter methylation was associated with a poor prognosis in stage II, microsatellite stability (MSS), BRAF wild-type (WT) CRCs: multivariate Cox proportional HR = 3.89 [95% confidence interval (CI), 1.58-9.60, P < 0.01; n = 66] and HR = 2.11 (95% CI, 0.95-4.69, P = 0.068, n = 136) in a second independent population-based study. CHFR promoter CpG island methylation, which is associated with MSI, also occurs frequently in MSS CRCs and is a promising prognostic marker in stage II, MSS, BRAF WT CRCs.


H2 clearance measurement of blood flow: a review of technique and polarographic principles.
H2 clearance is a powerful method for monitoring blood flow. Simple and inexpensive to implement, the method allows multiple in situ determinations of blood flow from any tissue in which a small electrode can be implanted. There is, however, evidence to suggest that H2 clearance is neither as accurate nor as local a measure of blood flow as generally supposed. Both in theory and practice, it probably cannot accurately determine blood flow rates greater than 100 ml/100 gm/min or localize blood flow to tissue volumes of less than 5 ml. Moreover, its experimental application is complicated by many technical problems hitherto largely ignored by workers in the field. Some of these problems arise from the limitations of the steady state polarographic technique used to measure tissue H2 concentrations. Other problems stem from the failure to consider possible sources of error in H2 clearance monitoring; these induce interference with the H2 signal by spurious electrode and tissue currents, and contributions from tissue ascorbate and O2. Nevertheless, with the appropriate safeguards and qualifications, H2 clearance is a valid and important approach to measuring blood


Substitution across professions within the home care sector: an investigation of nursing and allied health services.
The service type offered by a home care agency contracted by the Queensland government is not based on the qualifications of the worker providing the service, but the service itself. This allows agencies to substitute certain levels and categories of labour in order to provide a service to meet their contract obligations. This study investigated evidence of labour substitution between nursing and allied health services. Correlation and regression analysis was performed on the data collected from 218 clients of a branch of a community-based service agency operating nationally in Brisbane, Australia, during April, May and June 2005. The results of the regression analysis revealed that when either allied health or nursing time rose by 10%, all else held constant, it was predicted that the other would fall by 4%. The subcategories, registered/enrolled nursing and physiotherapy, appeared to drive the inverse relationship between nursing and allied health service time. Registered/enrolled nursing service time was more sensitive to changes in physiotherapy rather than the other way around. The higher labour turnover among allied health staff compared with the nursing staff reported by the agency implies a substitution of labour between the two professions to ensure that the needs of clients are met. Health policy makers and health care professional educators need to acknowledge that workforce shortages will inevitably reshape professional boundaries. Leaving labour-force substitution undiscussed and unplanned may compromise the quality and safety of care.


High-throughput high-resolution class I HLA genotyping in East Africa.
HLA, the most genetically diverse loci in the human genome, play a crucial role in host-pathogen interaction by mediating innate and adaptive cellular immune responses. A vast number of infectious diseases affect East Africa, including HIV/AIDS, malaria, and tuberculosis, but the HLA genetic diversity in this region remains incompletely described. This is a major obstacle for the design and evaluation of preventive vaccines. Available HLA typing techniques, that provide the 4-digit level resolution needed to interpret immune responses, lack sufficient throughput for large immunoepidemiological studies. Here we present a novel HLA typing assay bridging the gap between high resolution and high throughput. The assay is based on real-time PCR using sequence-specific primers (SSP) and can genotype carriers of the 49 most common East African class I HLA-A, -B, and -C alleles, at the 4-digit level. Using a validation panel of 175 samples from Kampala, Uganda, previously defined by sequence-based typing, the new assay performed with 100% sensitivity and specificity. The assay was also implemented to define the HLA genetic complexity of a previously uncharacterized Tanzanian population, demonstrating its inclusion in the major East African genetic cluster. The availability of genotyping tools with this capacity will be extremely useful in the identification of correlates of immune protection and the evaluation of candidate vaccine efficacy.


Atomically deviated Pd-Te nanoplates boost methanol-tolerant fuel cells.
The methanol crossover effect in direct methanol fuel cells (DMFCs) can severely reduce cathodic oxygen reduction reaction (ORR) performance and fuel efficiency. As a result, developing efficient catalysts with simultaneously high ORR activity and excellent antipoisoning methanol capability remains challenging. Here, we report a class of Pd-Te hexagonal nanoplates (HPs) with a Pd20Te7 phase that simultaneously overcome the activity and methanol-tolerant issues in alkaline DMFC. Because of the specific arrangement of Pd atoms deviated from typical hexagonal close-packing, Pd-Te HPs/C displays extraordinary methanol tolerance with high ORR performance compared with commercial Pt/C. DFT calculations reveal that the high performance of Pd-Te HPs can be attributed to the breakthrough of the linear relationship between OOH* and OH* adsorption, which leaves sufficient room to improve the ORR activity but suppresses the methanol oxidation reaction. The concurrent high ORR activity and excellent methanol tolerance endow Pd-Te HPs as practical electrocatalysts for DMFC and beyond.


Food allergy needs assessment, training curriculum, and knowledge assessment for child care.
More than half of preschool-aged children are enrolled in child care in the United States. Roughly 8% of children between 3 and 5 years of age have a food allergy. Child care center workers (CCCWs) are important caregivers who frequently encounter food allergies, but little is known about their education and understanding of food allergy and anaphylaxis. To perform a food allergy and anaphylaxis educational needs assessment, provide a training curriculum for CCCWs, and assess the effectiveness of the training curriculum. An online educational needs assessment and live training curriculum addressing food allergy recognition, treatment, and food labeling with pretests and posttests were created, and content and face validity were obtained. A needs assessment survey was sent to centers in Dallas and Tarrant counties. The training curriculum was performed at continuing education conferences. Seventy-three workers responded to the needs assessment, with 46% reporting prior food allergy training. They reported information sources as parents (73%), self (54%), educational curricula (21%), and conferences (19%). Most believed they have a high or moderately high proficiency in food allergy management. Forty-five workers participated in the training curriculum. Total scores improved from 54% correct on the pretest to 83% correct on the posttest (P < .001). Categorical subanalysis reveals similar results, with statistically significant improvement in all areas. CCCWs have diverse educational backgrounds and infrequently experience standardized training about food allergies. There is a significant lack of knowledge regarding food allergies and anaphylaxis. The curriculum was successful at increasing food allergy knowledge among CCCWs.


Mechanistic mathematical models: An underused platform for HPV research.
Health economic modeling has become an invaluable methodology for the design and evaluation of clinical and public health interventions against the human papillomavirus (HPV) and associated diseases. At the same time, relatively little attention has been paid to a different yet complementary class of models, namely that of mechanistic mathematical models. The primary focus of mechanistic mathematical models is to better understand the intricate biologic mechanisms and dynamics of disease. Inspired by a long and successful history of mechanistic modeling in other biomedical fields, we highlight several areas of HPV research where mechanistic models have the potential to advance the field. We argue that by building quantitative bridges between biologic mechanism and population level data, mechanistic mathematical models provide a unique platform to enable collaborations between experimentalists who collect data at different physical scales of the HPV infection process. Through such collaborations, mechanistic mathematical models can accelerate and enhance the investigation of HPV and related diseases.


Effect of ventilation technique and airway diameter on bronchial lumen to pulmonary artery diameter ratios in clinically normal beagle dogs.
In dogs, a mean broncho-arterial ratio of 1.45 ± 0.21 has been previously defined as normal. These values were obtained in dogs under general inhalational anesthesia using a single breath-hold technique. The purpose of the study was to determine whether ventilation technique and bronchial diameter have an effect on broncho-arterial ratios. Four healthy Beagle dogs were scanned twice, each time with positive-pressure inspiration and end expiration. For each ventilation technique, broncho-arterial ratios were grouped into those obtained from small or large bronchi using the median diameter of the bronchi as the cutoff value. Mean broncho-arterial ratios obtained using positive-pressure inspiration (1.24 ± 0.23) were statistically greater than those obtained at end expiration (1.11 ± 0.20) P = 0.005. There was a strong positive correlation between bronchial diameter and broncho-arterial ratios for both ventilation techniques (positive-pressure inspiration rs = .786, P < 0.0005 and end expiration rs = .709, P < 0.0005). Mean broncho-arterial ratio for the large bronchi obtained applying positive-pressure inspiration was 1.39 cm ± 0.20 and during end expiration was 1.22 cm ± 0.20. Mean broncho-arterial ratio for the small bronchi obtained during positive-pressure inspiration was 1.08 cm ± 0.13 and during end expiration was 1.01 cm ± 0.13. There was a statistically significant difference between these groups (F = 248.60, P = 0.005). Findings indicated that reference values obtained using positive-pressure inspiration or from the larger bronchi may not be applicable to dogs scanned during end expiration or to the smaller bronchi.


Unilateral keratoderma in a mother and her son.
Keratoderma is a group of conditions characterized by hyperkeratosis affecting the skin on the soles of the feet and palms of the hands bilaterally. The classification of keratodermas depends on whether it is inherited or acquired and on its clinical features, including diffuse or focal involvement of the skin and the morphology of lesions present. We describe the rare case of a 54-year-old female who presented with a nearly 40-year history of punctate keratoderma on her right palm and sole. History taking revealed that her biologic son also has unilateral left-sided keratoderma. The clinical presentation of unilateral keratoderma has been reported only four times in the literature.


Setting the scene: a scoping review of gambling research in Ghana.
Ghana, like many other African countries, has seen a rapid expansion in gambling industry activity in the last decade with sports betting becoming a popular pastime among young Ghanaian males. The proliferation of land based, and online gambling operations presents a significant public health threat to the citizenry. Yet little is known about gambling participation and harms, the size of the gambling market and the role of the gambling industry in influencing and perpetuating gambling behaviour in Ghana. The aim of this study was to contribute to understanding the extent of gambling research in Ghana by mapping out what the current literature indicates. Six databases were searched for peer-reviewed journal articles that focused on gambling in Ghana. The findings from this review demonstrate gambling research in Ghana is in a nascent state with a predominant focus on individual factors and limited consideration of public health. The implications for future research are highlighted, including the requirement for a nationally representative study to understand the prevalence of gambling participation and harm, and the role of the gambling industry. Public health action grounded in sound evidence is urgently required to address the individual and community level problems associated with this harmful commodity.


[Determination of the number of spermatozoa per 1 ml of ejaculate in a stained smear of the semen].
Using statistical and morphometric methods, the authors validate a new method for assessing the quantity of spermatozoa per ml of native ejaculate in a fixed and stained smear. The proposed method is regarded as an alternative to the routine technique.


Combining test-taking intervention with course remediation: effects on National Board subtest performance.
Effects of test-taking skills instruction over a five-year period were assessed by examining retest performances of second-year medical students who, because of unsatisfactory performance, were required to repeat National Board Part I subtests and who all participated in a summer review program. Two groups of students were involved in the study: an intervention group consisting of 14 students who participated in a special intervention program involving test-taking skills instruction and a comparison group that included 13 nonparticipants. The intervention group's mean retest performance was significantly greater (F = 9.38, P < .005). The results suggest that intervention programs can have a significant impact on academic success, and deserve serious consideration by medical students who demonstrate low performances on examinations because of comparatively poor test-taking skills and for students undergoing academic remediation.


[Assessment of time-dependent effects of long-acting altiazem in patients with hypertension stage II].
A chronodiagnostic examination covered 27 patients with hypertension stage II of indifferent chronotype. The patients were divided into 3 groups by the time of intake of a single daily dose of altiazem--at 8 a.m., 14 p.m., 20 p.m. Chronological organization of the cardiovascular system was studied with autorhythmometry for 3 days in 4-h intervals including night before and in the end of the treatment. Echo-CG, doppler echocardiography and rheoencephalography investigated hemodynamics once daily also before and in the end of the treatment. The findings show advantage of chronotherapeutic prescription of altiazem PP in a mean dose 220 mg/day at evening hours in non-chronodependent patients with hypertension stage II because such a regimen brought about more adequate restoration of the cardiovascular system time organization both by the spectrum of significant rhythms and their parameters; by better improvement of left ventricular diastolic function and pulse cerebral flow. Faster achievement of the therapeutic effect in the evening intake of altiazem supports its chronotherapeutic administration. This leads to reduction of the drug course dose and, therefore, treatment duration.


Postoperative changes in cerebral metabolism in temporal lobe epilepsy.
Fludeoxyglucose F 18 positron emission tomography ((18)F-FDG-PET) can detect focal metabolic abnormalities ipsilateral to the seizure focus in 80% of patients with temporal lobe epilepsy (TLE). Regions outside the epileptogenic zone can also be affected. We hypothesized that these remote regions might show altered metabolism, tending to return toward normal values, after surgery. Interictal preoperative and postoperative (18)F-FDG-PET metabolism were compared in patients with refractory TLE. Based on pathological findings, disease was classified in the following 3 groups: mesial temporal sclerosis, mass lesions, and no pathological diagnosis. Quantitative PET data analysis was performed using the region-of-interest template previously described. Global normalization was used to adjust for the effect of antiepileptic medication changes. Data were analyzed by Wilcoxon signed rank test and analysis of variance. The Clinical Epilepsy Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health. Twenty-two patients with refractory TLE. Preoperatively, in all groups, cerebral metabolic rate for glucose was decreased ipsilateral to the resection site in inferior lateral temporal, inferior mesial temporal, and inferior frontal areas and thalamus. Postoperatively, in all groups, cerebral metabolic rate for glucose increased in ipsilateral inferior frontal area and thalamus. In the mesial temporal sclerosis group, we found a statistically significant increase in the contralateral thalamus. Temporal lobe epilepsy is associated with extensive preoperative decreased metabolism in inferior lateral temporal, inferior mesial temporal, and inferior frontal areas and thalamus. Postoperatively, we found increased IF and thalamic metabolism. Seizures may have a reversible effect on brain areas connected with, but remote from, the epileptogenic cortex. Arch Neurol. 2000;57:1447-1452


ER stress and the unfolded protein response in neurodegeneration.
The clinical manifestation of neurodegenerative diseases is initiated by the selective alteration in the functionality of distinct neuronal populations. The pathology of many neurodegenerative diseases includes accumulation of misfolded proteins in the brain. In physiological conditions, the proteostasis network maintains normal protein folding, trafficking and degradation; alterations in this network - particularly disturbances to the function of endoplasmic reticulum (ER) - are thought to contribute to abnormal protein aggregation. ER stress triggers a signalling reaction known as the unfolded protein response (UPR), which induces adaptive programmes that improve protein folding and promote quality control mechanisms and degradative pathways or can activate apoptosis when damage is irreversible. In this Review, we discuss the latest advances in defining the functional contribution of ER stress to brain diseases, including novel evidence that relates the UPR to synaptic function, which has implications for cognition and memory. A complex concept is emerging wherein the consequences of ER stress can differ drastically depending on the disease context and the UPR signalling pathway that is altered. Strategies to target specific components of the UPR using small molecules and gene therapy are in development, and promise interesting avenues for future interventions to delay or stop neurodegeneration.


Antinociceptive and Anti-inflammatory Effects of Pistacia vera LeafExtract in Mice.
Pistacia vera L., a member of Anacardiaceae family, has been used for sedation and analgesia in traditional medicine. In this study, the antinociceptive and anti-inflammatory effects as well as acute toxicity of the aqueous and ethanolic extracts of P. vera leaves were investigated in mice. The antinociceptive activity was studied using hot plate and writhing tests. The effect of the extracts against acute inflammation was determined using xylene-induced ear edema and the activity of the extracts, against chronic inflammation, was assessed using the cotton pellet test. The LD50 values of the infusion and maceration extracts were 0.8 g/Kg and 0.79 g/Kg, respectively. The aqueous and ethanolic maceration extracts of the P. vera leaves at the doses of 0.4 g/Kg and 0.5 g/Kg (IP), respectively, showed antinociceptive effects. The pretreatment of naloxone (2 mg/Kg, SC) inhibited the activities of extracts in hot plate test, but naloxone at the same dose could not inhibit the antinociceptive activity in writhing test. The extracts also showed anti-inflammatory effects in acute and chronic anti-inflammatory tests. The ethanolic extract was as effective as diclofenac in both inflammatory tests. The aqueous and ethanolic extracts of P. vera leaves demonstrated central and peripheral antinociceptive activities dose-dependently and the central effect may be mediated by opioid system. The extracts also demonstrated anti-inflammatory effects against acute and chronic inflammation.


[The management in clinical trials of pharmacological agents].
The author analyzes the experience gained by foreign countries in the creation of a new management system in all spheres of public health, including clinical trials and use of drugs. A term "High-Quality Clinical Practice" is used in the European Community, reflecting the standard or the norm of clinical studies and designed as a complex of regulations for the organization and implementation of clinical studies. High-Quality Clinical Practice implies all reasonable measures providing the accuracy of experimental data and guarantees the rights of the participants in the trials. Studies carried out in conformity with the requirements of High-Quality Clinical Practice bring about reliable results and permit the pharmaceutical companies and public health organs use these data.


Pharmacological dissection of pyloric network of the lobster stomatogastric ganglion using picrotoxin.
1. Picrotoxin (PTX) (10(-7)-10(-6) M) completely blocked most inhibitory synapses in the pyloric pattern generator of the lobster (Panulirus interruptus) stomatogastric ganglion. The sensitivity of synapses from most classes of identified neurons was examined. Blockade was at least partly reversible with prolonged washing. 2. The synapses from pyloric dilator (PD) neurons were the only inhibitory synapses that picrotoxin failed to block completely. 3. A correlation is derived that brief, fast-rise inhibitory postsynaptic potentials (IPSPs) are picrotoxin sensitive, whereas a slow rounded component of IPSPs from PD neurons is not picrotoxin sensitive. 4. Picrotoxin caused specific changes in the pattern of the motor rhythm produced by the 16-cell pyloric network. This sheds some light on the functional role of particular synapses in the pyloric generator. 5. The endogenously bursting neurons (PD and anterior burster (AB)), which drive the pyloric rhythm, kept a similar burst rate. 6. Under picrotoxin, the pyloric "follower" neurons all moved to later phase relative to the "driver" group. Some normally antagonistic cells, related by reciprocal inhibitor connections, became in-phase. These and other pattern changes could be related to blockade of particular synapses. 7. The pyloric rhythm was still quite recognizable under picrotoxin despite the drastically altered circuitry of the synaptic network. This supports the idea that periodic inhibition from the PD driver neurons plays a primary role in creating the pyloric pattern.


Animal health and greenhouse gas intensity: the paradox of periparturient parasitism.
Here we provide the first known direct measurements of pathogen challenge impacts on greenhouse gas production, yield and intensity. Twin-rearing ewes were ad libitum fed pelleted lucerne from day -32 to 36 (day 0 is parturition), and repeatedly infected with 10,000 Teladorsagia circumcincta infective larvae (n=16), or sham-dosed with water (n=16). A third group of 16 ewes were fed at 80% of uninfected ewes' feed intake during lactation. Methane emissions were measured in respiration chambers (day 30-36) whilst total tract apparent nutrient digestibility around day 28 informed calculated manure methane and nitrous oxide emissions estimates. Periparturient parasitism reduced feed intake (-9%) and litter weight gain (-7%) and doubled maternal body weight loss. Parasitism reduced daily enteric methane production by 10%, did not affect the methane yield per unit of dry matter intake but increased the yield per unit of digestible organic matter intake by 14%. Parasitism did not affect the daily calculated manure methane and nitrous oxide production, but increased the manure methane and nitrous oxide yields per unit of dry matter intake by 16% and 4%, respectively, and per unit of digestible organic matter intake by 46% and 31%, respectively. Accounting for increased lucerne input for delayed weaning and maternal body weight loss compensation, parasitism increased the calculated greenhouse gas intensity per kg of lamb weight gain for enteric methane (+11%), manure methane (+32%) and nitrous oxide (+30%). Supplemented with the global warming potential associated with production of pelleted lucerne, we demonstrated that parasitism increased calculated global warming potential per kg of lamb weight gain by 16%, which was similar to the measured impact of parasitism on the feed conversion ratio. Thus, arising from a pathogen-induced feed efficiency reduction and modified greenhouse gas emissions, we demonstrated that ovine periparturient parasitism increases greenhouse gas intensity. This implies that ewe worm control can not only improve production efficiency but also reduce the environmental footprint of sheep production systems.


Exciplex Organic Light-Emitting Diodes with Nearly 20% External Quantum Efficiency: Effect of Intermolecular Steric Hindrance between the Donor and Acceptor Pair.
Exciplex emitters have emerged as an important class of thermally activated delayed fluorescence (TADF) materials for highly efficient OLEDs. A TADF exciplex emitter requires an intermolecular donor/acceptor pair. We have synthesized a bipolar donor-type material, DPSTPA, which was used to pair with known acceptor materials (2CzPN, 4CzIPN, or CzDBA). The OLEDs based on the exciplex emitters, DPSTPA/X, where X = 2CzPN and CzDBA, give green and orange-red colors with record-high external quantum efficiencies (EQEs) of 19.0 ± 0.6 and 14.6 ± 0.4%, respectively. In contrast, the exciplex pair DPSTPA/4CzIPN gave a very low photoluminescence quantum yield (PLQY) and a very low EQE value of the device. The DFT calculations indicate that the intermolecular distance between the donor and the acceptor plays a key factor for the PLQY and EQE. The observed low PLQY and the poor device performance for the DPSTPA/4CzIPN pair are probably because of the relatively long distance between the DPSTPA and 4CzIPN in the thin film caused by the four congested carbazole (Cz) groups of 4CzIPN, which effectively block the interaction of the nitrile acceptor with the triphenylamino donor of DPSTPA.