Serum total testosterone is a significant preoperative variable independently contributing to separating the prostate cancer population into prostatectomy Gleason score groups.
To investigate the potential of preoperative serum total testosterone (TT) in contributing to the definition of separate prostatectomy Gleason score (pGS) groups of the prostate cancer (PCa) population. The data of 220 patients operated on for PCa were retrospectively reviewed. No patient had previously received 5α-reductase inhibitor, luteinizing hormone-releasing analogs or testosterone replacement treatment. The patient population was grouped according to the pGS as 6 = 3+3, 7 = 3+4, 7 = 4+3 and 8-10. Eight variables were simultaneously investigated in each group: prostate-specific antigen (PSA), TT, free testosterone, age, percentage of positive prostate biopsy cores (P+), biopsy Gleason score (bGS), overall cancer volume estimated as percentage of prostate volume (V+) and prostate weight (Wi). Univariate analysis of variance (ANOVA), multivariate analysis of variance (MANOVA) and multivariate discriminant analysis (MDA) were the statistical methods used for evaluating the data. There were 89 patients in pGS 6 = 3+3, 84 in pGS 7 = 3+4, 24 in pGS 7 = 4+3 and 23 in pGS 8-10. ANOVA showed that bGS (p < 0.0001), P+ (p < 0.0001), V+ (p < 0.0001), PSA (p = 0.0001), Wi (p = 0.0002) and TT (p = 0.01) were significantly different in the four pGS groups. MANOVA tests showed that only bGS (p < 0.0001), V+ (p = 0.0003), TT (p = 0.001) and, to a lesser extent, PSA (p = 0.06) were the significant variables that individually and independently contributed a significant amount to separation of the four pGS groups of the PCa population. MDA showed that the independent variables ranked as bGS (p < 0.0001), TT (p = 0.001), V+ (p = 0.001) and PSA (p = 0.06). Serum TT is a significant preoperative variable that independently contributes to separating the PCa population into pGS score groups. Pretreatment baseline serum TT levels should be measured and their inclusion in neural networks predicting PCa natural history be considered in the patient population diagnosed with PCa.


Effect of a heavy heteroatom on triplet formation and interactions in single conjugated polymer molecules and aggregates.
Triplet formation and interactions with emissive singlet excitons are investigated in poly(3-hexylselenophene) (P3HS) using single molecule spectroscopy. P3HS is a heavy atom analog of the more commonly studied poly(3-hexylthiophene) (P3HT), a benchmark polymer for solar cells. P3HS tends to aggregate strongly which necessitates dilution to ultra-low levels within a solid inert host in order to resolve photophysical responses of single chains. Fluorescence excitation intensity modulation is performed on isolated P3HS chains using a sequence of rectangular pulses of varying intensities to probe the presence of spin-forbidden triplet excitons. Triplet population dynamics originating from singlet-triplet and triplet-triplet interactions appear as quenching of the initial fluorescence intensity to steady-state levels on characteristic time scales of ∼1-10 μs. Over 80% of all molecules studied display significant fluorescence intensity modulation (quenching depths >50%) indicative of efficient intersystem crossing and large triplet occupancies. Because triplets are highly localized and singlet-triplet and triplet-triplet annihilation rate constants are comparable to those of intersystem crossing, multiple triplets are present at any given time on single P3HS chains. Triplet lifetimes were estimated to be ∼4 μs (upper limit) determined from recovery to the ground electronic singlet state in the absence of light and, surprisingly, triplets vanish at the onset of P3HS aggregation. This result was unexpected since P3HS triplet formation takes place on time scales <30 ps making this process competitive with most accessible non-radiative deactivation pathways.


Non-invasive assessment of aortic stenosis by Doppler ultrasound.
The peak pressure drop across the aortic valve in aortic stenosis has been measured by Doppler ultrasound. Maximum velocity in the Doppler signal from the aortic jet was recorded using a maximum frequency estimator. With an angle close to zero between ultrasound beam and maximal velocity in the jet, peak pressure drop can be calculated from the maximal velocity measured; a larger angle will underestimate maximal velocity and pressure drop. In 57 of 63 patients with aortic stenosis, the aortic jet could be reached by the ultrasound beam and, in 37 of these, peak pressure drop by ultrasound was compared with that obtained at catheterisation. In patients less than 50 years of age the aortic jet was easy to find, the measurement was reproducible, and underestimation of the pressure drop obtained at catheterisation was within 25 per cent in 17 of 18 patients. In patients over 50 years Doppler signals from the aortic jet were more difficult to obtain, and pressure drop was significantly underestimated in one-third, but time of maximum velocity in systole could indicate whether moderate or severe aortic stenosis was present.


Vaccination-induced autoimmune vitiligo is a consequence of secondary trauma to the skin.
A major concern for cancer vaccines targeting self-tumor antigens is the risk of autoimmune sequelae. Although antitumor immunity correlates with autoimmune disease in some preclinical models, the mechanism(s) linking antitumor immunity and subsequent autoimmune pathology remain(s) to be determined. In the current study, we demonstrated that intradermal (i.d.) immunization with a recombinant adenovirus (Ad) expressing the murine melanoma antigen tyrosinase-related protein 2 (AdmTrp-2) results in a moderate level of tumor protection against the B16F10 murine melanoma without any vitiligo. Similar immunization with an Ad encoding human Trp-2 (AdhTrp-2) resulted in 50-fold greater protective immunity and produced vitiligo in all of the mice, suggesting that the development of autoimmunity may reflect the potency of the vaccine. Interestingly, delivery of AdhTrp-2 by i.m. injection generated protective immunity comparable with that seen in mice that received the vaccine by the i.d. route, but none of the recipients in the i.m. group developed vitiligo. The cellular and humoral responses in the i.m. immunized mice were greater than in the i.d. group; therefore, the lack of vitiligo was not caused by reduced efficacy of the vaccine. These results led us to hypothesize that vaccine-induced vitiligo was associated with local inflammatory responses. Mice immunized i.m. with AdhTrp-2 did develop vitiligo when they subsequently were injected i.d. with either a control Ad vector or complete Freund's adjuvant, suggesting that vitiligo is initiated by some form of trauma within the skin. Our data demonstrated that autoimmune pathology is not an unavoidable outcome of effective cancer vaccines directed against self-tumor antigens.


Using out-of-office blood pressure measurements in established cardiovascular risk scores: a secondary analysis of data from two blood pressure monitoring studies.
Blood pressure (BP) measurement is increasingly carried out through home or ambulatory monitoring, yet existing cardiovascular risk scores were developed for use with measurements obtained in clinics. To describe differences in cardiovascular risk estimates obtained using ambulatory or home BP measurements instead of clinic readings. Secondary analysis of data from adults aged 25-84 years in the UK and the Netherlands without prior history of cardiovascular disease (CVD) in two BP monitoring studies: the Blood Pressure in different Ethnic groups (BP-Eth) study and the Home versus Office blood pressure MEasurements: Reduction of Unnecessary treatment Study (HOMERUS). The primary comparison was Framingham risk calculated using BP measured as in the Framingham study or daytime ambulatory BP measurements. Statistical significance was determined using non-parametric tests. In 442 BP-Eth patients (mean age = 58 years, 50% female [n = 222]) the median absolute difference in 10-year Framingham cardiovascular risk calculated using BP measured as in the Framingham study or daytime ambulatory BP measurements was 1.84% (interquartile range [IQR] 0.65-3.63, P = 0.67). In 165 HOMERUS patients (mean age = 56 years, 46% female) the median absolute difference in 10-year risk for daytime ambulatory BP was 2.76% (IQR 1.19-6.39, P<0.001) and only 8 out of 165 (4.8%) of patients were reclassified. Estimates of cardiovascular risk are similar when calculated using BP measurements obtained as in the risk score derivation study or through ambulatory monitoring. Further research is required to determine if differences in estimated risk would meaningfully influence risk score accuracy.


Mental Health and School Functioning for Girls in the Child Welfare System: the Mediating Role of Future Orientation and School Engagement.
This study investigated the association between mental health problems and academic and behavioral school functioning for adolescent girls in the child welfare system and determined whether school engagement and future orientation meditated the relationship. Participants were 231 girls aged between 12 and 19 who had been involved with the child welfare system. Results indicated that 39% of girls reported depressive symptoms in the clinical range and 54% reported posttraumatic symptoms in the clinical range. The most common school functioning problems reported were failing a class (41%) and physical fights with other students (35%). Participants reported a mean number of 1.7 school functioning problems. Higher levels of depression and PTSD were significantly associated with more school functioning problems. School engagement fully mediated the relationship between depression and school functioning and between PTSD and school functioning, both models controlling for age, race, and placement stability. Future orientation was not significantly associated with school functioning problems at the bivariate level. Findings suggest that school engagement is a potentially modifiable target for interventions aiming to ameliorate the negative influence of mental health problems on school functioning for adolescent girls with histories of abuse or neglect.


Overview lecture. Hydration processes in biological and macromolecular systems.
I discuss several themes arising from the papers presented at this Faraday Discussion that are, in my view, particularly interesting and/or important. With respect to model systems, recent progress in understanding structural aspects of the hydrophobic interaction, and of through-solvent interactions in general, is highlighted, together with the need to continue to develop more sophisticated (theoretical and computational) interpretational techniques if we are to exploit to the full the power of present-day experimental techniques. The current state of our knowledge of hydration effects on the structure and dynamics of biomolecules is discussed, and the importance of being able to see how molecular-level structural effects control behaviour at the important mesoscopic level is underlined. Issues relating to recent progress in characterising solvent effects in more complex systems and processes, including those of industrial interest, are raised, and the necessity of using a range of appropriate experimental techniques when tackling such complex problems is stressed. Progress since the 1975 Royal Society Discussion is highlighted, and interesting issues ripe for fruitful discussion at this meeting are raised. A strong case can now be made that our under-standing of both the structure and dynamics of water as a function of its local environment is now sufficiently good to enable us to use water as a probe of complex system behaviour, rather than, as heretofore, an objects of study in itself.


A carving method to determine an optimal working projection using three-dimensional volume rendering digital subtraction angiography in coil embolization of cerebral aneurysms.
A working projection in coil embolization of a cerebral aneurysm is usually determined using three-dimensional volume rendering digital subtraction angiography (3D VRDSA), in which the boundary between an aneurysm and its parent artery cannot be identified simultaneously on the front and back surfaces of the vessel. A new method was devised to obtain a more optimal working projection. 83 patients with aneurysms being treated by embolization were retrospectively evaluated. An aneurysm was accurately distinguished from the parent artery by observation of front, back, proximal and distal sides of the vessel on 3D VRDSA (carving method). An optimal working projection with simultaneous identification of the front and back boundary lines was determined using a translucent vessel complex combined with the carved aneurysm. In 32 aneurysms (38.6%), the optimal working projection was consistent with the working projection that had been used during the procedure. In terminal type aneurysms, the angle difference between the optimal and actual working projections was significantly smaller than in the other types (p<0.05). Aneurysms with a maximal diameter <5 mm showed a significantly larger angle difference between the optimal and actual working projections than aneurysms with a maximal diameter ≥5 mm (p<0.05). In more than half of the patients, the actual working projection was inaccurate. The carving method might be useful to determine working projections, especially for aneurysms other than the terminal type and/or those with a maximal diameter <5 mm.


Recombination at mammalian telomeres: an alternative mechanism for telomere protection and elongation.
In human cells, homologous recombination (HR) provides an accurate mechanism for the repair of DNA double-strand breaks caused by replication fork breakdown or DNA damaging agents. HR also plays a role in the maintenance of eukaryotic telomeres; cells defective in the recombinational repair proteins RAD51D or RAD54 exhibit telomere shortening and end-to-end chromosome fusions. Here we discuss the way in which HR contributes to telomere protection and elongation in mammalian cells. Understanding the mechanisms by which HR promotes telomere maintenance has important implications for genomic stability and tumorigenesis.


Using Digital Pathology to Understand Epithelial Characteristics of Benign Breast Disease among Women Undergoing Diagnostic Image-Guided Breast Biopsy.
Delayed terminal duct lobular unit (TDLU) involution is associated with elevated mammographic breast density (MD). Both are independent breast cancer risk factors among women with benign breast disease (BBD). Prior digital analyses of normal breast tissues revealed that epithelial nuclear density (END) and TDLU involution are inversely correlated. Accordingly, we examined associations of END, TDLU involution, and MD in BBD clinical biopsies. This study included digitized images of 262 representative image-guided hematoxylin and eosin-stained biopsies from 224 women diagnosed with BBD, enrolled within the cross-sectional BREAST-Stamp project that were visually assessed for TDLU involution (TDLU count/100 mm2, median TDLU span and median acini count per TDLU). A digital algorithm estimated nuclei count per unit epithelial area, or END. Single X-ray absorptiometry of prebiopsy ipsilateral craniocaudal digital mammograms measured global and localized MD surrounding the biopsy region. Adjusted ordinal logistic regression models assessed relationships between tertiles of TDLU and END measures. Analysis of covariance examined mean differences in MD across END tertiles. TDLU measures were positively associated with increasing END tertiles [TDLU count/100 mm2, ORT3vsT1: 3.42, 95% confidence interval (CI), 1.87-6.28; acini count/TDLUT3vsT1, OR: 2.40, 95% CI, 1.39-4.15]. END was significantly associated with localized, but not, global MD. Relationships were most apparent among patients with nonproliferative BBD. These findings suggest that quantitative END reflects different but complementary information to the histologic information captured by visual TDLU and radiologic MD measures and merits continued evaluation in assessing cellularity of breast parenchyma to understand the etiology of BBD.


A modelling analysis of a new multi-stage pathway for classifying achievement of public health milestones for leprosy.
Several countries have come close to eliminating leprosy, but leprosy cases continue to be detected at low levels. Due to the long, highly variable delay from infection to detection, the relationship between observed cases and transmission is uncertain. The World Health Organization's new technical guidance provides a path for countries to reach elimination. We use a simple probabilistic model to simulate the stochastic dynamics of detected cases as transmission declines, and evaluate progress through the new public health milestones. In simulations where transmission is halted, 5 years of zero incidence in autochthonous children, combined with 3 years of zero incidence in all ages is a flawed indicator that transmission has halted (54% correctly classified). A further 10 years of only occasional sporadic cases is associated with a high probability of having interrupted transmission (99%). If, however, transmission continues at extremely low levels, it is possible that cases could be misidentified as historic cases from the tail of the incubation period distribution, although misleadingly achieving all three milestones is unlikely (less than 1% probability across a 15-year period of ongoing low-level transmission). These results demonstrate the feasibility and challenges of a phased progression of milestones towards interruption of transmission, allowing assessment of programme status. This article is part of the theme issue 'Challenges and opportunities in the fight against neglected tropical diseases: a decade from the London Declaration on NTDs'.


Beta adrenergic modulation of formyl-methionine-leucine-phenylalanine-stimulated secretion of eosinophil peroxidase and leukotriene C4.
The inhibitory effect of beta-2 adrenergic receptor stimulation on leukotriene C4 (LTC4) secretion and eosinophil peroxidase (EPO) release caused by exogenous activation with 10(-8) to 10(-6) M formyl-met-leu-phe (fMLP) + 5 micrograms/ml of cytochalasin B (Cyto B) in purified human peripheral blood eosinophils was studied. Cells from normal subjects were isolated by negative immunoselection and remained > or = 98% viable as determined by trypan blue exclusion. Duplicate aliquots of eosinophils (10(5) cells/intervention) were activated with 1) fMLP + Cyto B alone, 2) fMLP + Cyto B after pretreatment with 10(-8) M albuterol, 3) 10(-8) M albuterol + fMLP + Cyto B after pretreatment with 10(-8) M propranolol or 4) vehicle control. After incubation, the supernatants were tested for concentration of LTC4 and EPO. Concentration-related release of EPO was demonstrated for 10(-8) M fMLP + 5 micrograms/ml of Cyto B to 10(-6) M fMLP + 5 micrograms/ml of Cyto B, and the greatest concentration of fMLP was used in all subsequent studies. FMLP + Cyto B caused substantial LTC4 secretion in eosinophils (300 +/- 83.0 pg/ml) as compared to sham-activated eosinophils (3.3 +/- 1.9 pg/ml; P < .02). Similarly, maximum EPO release increased from 277 +/- 17.8 to 3956 +/- 1230 ng/10(6) cells (P < .02) after activation with fMLP + Cyto B. Treatment with albuterol decreased markedly both LTC4 secretion to 144 +/- 54.0 pg/ml (P < .05 vs. fMLP + Cyto B-activated eosinophils) and EPO release to 1993 +/- 368 ng/10(6) cells (P < .05 vs. fMLP + Cyto B-activated eosinophils).(ABSTRACT TRUNCATED AT 250 WORDS)


Vitamin E supplementation increases T helper 1 cytokine production in old mice infected with influenza virus.
Compared with young mice, old mice infected with influenza virus have significantly higher pulmonary viral titres, although these can be reduced significantly with dietary vitamin E supplementation. T helper 1 (Th1) cytokines, especially interferon-gamma (IFN-gamma), play an important role in defending against influenza infection. However, there is an age-associated loss of Th1 cytokine production. Prostaglandin E2 (PGE2) production, which increases with age, can modulate the T helper cell function by suppressing Th1 cytokine production. To investigate the mechanism of vitamin E supplementation on reduction of influenza severity in old mice, we studied the cytokine production by splenocytes, and PGE2 production by macrophages (Mphi), in young and old C57BL mice fed semipurified diets containing 30 (control) or 500 parts per million (ppm) (supplemented) vitamin E for 8 weeks, and then infected with influenza A/PC/1/73 (H3N2). Old mice fed the control diet had significantly higher viral titres than young mice; old mice fed the vitamin E-supplemented diet had significantly lower pulmonary viral titres than those fed the control diet (P = 0.02 and 0.001 for overall age and diet effect, respectively). Following influenza infection, interleukin (IL)-2 and IFN-gamma production was significantly lower in old mice than in young mice. Vitamin E supplementation increased production of IL-2 and IFN-gamma in old mice; higher IFN-gamma production was associated with lower pulmonary viral titre. Old mice fed the control diet showed significantly higher lipopolysaccharide (LPS)-stimulated Mphi PGE2 production than old mice fed the vitamin E diet or young mice fed either diet. There was no significant age difference in IL-6, IL-1beta, or tumour necrosis factor-alpha (TNF-alpha) production by splenocytes. Young mice fed the vitamin E-supplemented diet had significantly lower IL-1beta (day 7) and TNF-alpha production (day 5) compared with those fed the control diet. Old mice fed the vitamin E-supplemented diet had significantly lower TNF-alpha production (day 2) than those fed the control diet. Our results indicate that the vitamin E-induced decrease in influenza viral titre is mediated through enhancement of Th1 cytokines, which may be the result of reduced PGE2 production caused by vitamin E.


Sensitization and chronic beryllium disease at a primary manufacturing facility, part 3: exposure-response among short-term workers.
Exposure-response relations for beryllium sensitization (BeS) and chronic beryllium disease (CBD) using aerosol mass concentration have been inconsistent, although process-related risks found in most studies suggest that exposure-dependent risks exist. We examined exposure-response relations using personal exposure estimates in a beryllium worker cohort with limited work tenure to minimize exposure misclassification. The population comprised workers employed in 1999 with six years or less tenure. Each completed a work history questionnaire and was evaluated for immunological sensitization and CBD. A job-exposure matrix was combined with work histories to create individual estimates of average, cumulative, and highest-job-worked exposure for total, respirable, and submicron beryllium mass concentrations. We obtained odds ratios from logistic regression models for exposure-response relations, and evaluated process-related risks. Participation was 90.7% (264/291 eligible). Sensitization prevalence was 9.8% (26/264), with 6 sensitized also diagnosed with CBD (2.3%, 6/264). A general pattern of increasing sensitization prevalence was observed as exposure quartile increased. Both total and respirable beryllium mass concentration estimates were positively associated with sensitization (average and highest job), and CBD (cumulative). Increased sensitization prevalence was identified in metal/oxide production, alloy melting and casting, and maintenance, and for CBD in melting and casting. Lower sensitization prevalence was observed in plant-area administrative work. Sensitization was associated with average and highest job exposures, and CBD was associated with cumulative exposure. Both total and respirable mass concentrations were relevant predictors of risk. New process-related risks were identified in melting and casting and maintenance.


Mixed Lactobacillus plantarum Strains Inhibit Staphylococcus aureus Induced Inflammation and Ameliorate Intestinal Microflora in Mice.
Objective. Staphylococcus aureus is an important pathogen that causes intestinal infection. We examined the immunomodulatory function of single and mixed Lactobacillus plantarum strains, as well as their impacts on the structure of the microbiome in mice infected with Staphylococcus aureus. The experiment was divided into three groups: protection, treatment, and control. Serum IFN-γ and IL-4 levels, as well as intestinal sIgA levels, were measured during and 1 week after infection with Staphylococcus aureus with and without Lactobacillus plantarum treatment. We used 16s rRNA tagged sequencing to analyze microbiome composition. IFN-γ/IL-4 ratio decreased significantly from infection to convalescence, especially in the mixed Lactobacillus plantarum group. In the mixed Lactobacillus plantarum group the secretion of sIgA in the intestine of mice (9.4-9.7 ug/mL) was significantly higher than in the single lactic acid bacteria group. The dominant phyla in mice are Firmicutes, Bacteroidetes, and Proteobacteria. Treatment with mixed lactic acid bacteria increased the anti-inflammatory factor and the secretion of sIgA in the intestine of mice infected with Staphylococcus aureus and inhibited inflammation.


Survey of 3-alkyl-2-methoxypyrazine content of South African Sauvignon blanc wines using a novel LC-APCI-MS/MS method.
An LC-MS/MS method for the trace-level determination of 3-alkyl-2-methoxypyrazines in Sauvignon blanc wines is described. 3-Isobutyl-2-methoxypyrazine (IBMP), 3-isopropyl-2-methoxypyrazine (IPMP) and 3-sec-butyl-2-methoxypyrazine (SBMP) were analyzed by reversed phase liquid chromatography coupled to atmospheric pressure chemical ionization, as electrospray ionization was found to suffer from matrix quenching effects. A sample preparation method involving distillation of wine followed by solvent extraction and sufficient preconcentration was developed. Limits of detection and quantification for all three analytes were 0.03 ng/L and 0.10 ng/L, respectively, making the method more sensitive than gas chromatographic methods. IBMP was found to be the most abundant congener in South African Sauvignon blanc wines, with concentrations varying between 0.40 and 44 ng/L in 575 samples. IPMP and SBMP levels varied from <0.03 to 3.9 and <0.03 to 3.2 ng/L, respectively. Statistical investigation indicated no clear correlation between methoxypyrazine content and either geographical origin or vintage. The method was also successfully applied for the quantitation of IBMP in five additional South African wine varieties, including three red wine cultivars. The developed method represents a powerful new tool for the in-depth investigation of these important wine aroma constituents.


Structural brain development: A review of methodological approaches and best practices.
Continued advances in neuroimaging technologies and statistical modelling capabilities have improved our knowledge of structural brain development in children and adolescents. While this has provided an increasingly nuanced understanding of brain development, the field is still plagued by inconsistent findings. This review highlights the methodological diversity in existing longitudinal magnetic resonance imaging (MRI) studies on structural brain development during childhood and adolescence, and addresses how such variation might contribute to inconsistencies in the literature. We discuss the impact of method choices at multiple decision points across the research process, from study design and sample selection, to image processing and statistical analysis. We also highlight the extent to which different methodological considerations have been empirically examined, drawing attention to specific areas that would benefit from future investigation. Where appropriate, we recommend certain best practices that would be beneficial for the field to adopt, including greater completeness and transparency in reporting methods, in order to ultimately develop an accurate and detailed understanding of normative child and adolescent brain development.


Hyperglycemia and Cancer: A State-of-the-Science Review.
Hyperglycemia can increase the risk for adverse events and outcomes in patients undergoing treatment for cancer. The purposes of this state-of-the-science review were to explore the complexity of hyperglycemia in patients with cancer and to analyze physiologic mechanisms and outcomes in individuals with or at risk for cancer. PubMed® and the Cochrane Library databases were searched, and 95 articles were included. Findings were evaluated for their methods and analyses. Studies assessed as methodologically flawed were not included. The synthesis of the articles provided the evidence for describing normal and glycemic pathways. Hyperglycemia in patients with cancer was explored through chronic inflammatory mechanisms that lead to increased risks for adverse events and outcomes. This article discusses normal glucose regulation and hyperglycemic pathways, hyperglycemia in patients with cancer, hyperglycemia and cancer-related inflammation, and outcomes (e.g., infections, mortality, symptoms). Understanding the contributors to and consequences of hyperglycemia can guide the development of screening tools to predict which individuals are at the greatest risk for hyperglycemic episodes prior to starting cancer therapies. Research can lead to glycemic guidelines specific to patients with cancer for better outcomes.


Detection of amplified DNA sequences by reverse chromosome painting using genomic tumor DNA as probe.
A modification of "reverse chromosome painting" was carried out using genomic DNA from tumor cells as a complex probe for chromosomal in situ suppression hybridization to normal metaphase chromosome spreads. Amplified DNA sequences contained in such probes showed specific signals, revealing the normal chromosome positions from which these sequences were derived. As a model system, genomic DNAs were analyzed from three tumor cell lines with amplification units including the proto-oncogene c-myc. The smallest amplification unit was about 90 kb and was present in 16-24 copies; the largest unit was bigger than 600 kb and was present in 16-32 copies. Specific signals that co-localized with a differently labeled c-myc probe on chromosome band 8q24 were obtained with genomic DNA from each cell line. In further experiments, genomic DNA derived from primary tumor material was used in the case of a male patient with glioblastoma multiforme (GBM). Southern blot analysis using an epidermal growth factor receptor gene (EGFR) probe that maps to 7p13 indicated the amplification of sequences from this gene. Using reverse chromosome painting, signals were found both on band 7p13 and bands 12q13-q15. Notably, the signal on 12q13-q15 was consistently stronger. The weaker 7p13 signal showed co-localization with the major signal of the differently labeled EGFR probe. A minor signal of this probe was seen on 12q13, suggesting cross-hybridization to ERB3 sequences homologous to EGFR. The results indicate co-amplification of sequences from bands 12q13-q15, in addition to sequences from band 7p13. Several oncogenes map to 12q13-q15 providing candidate genes for a tumor-associated proto-oncogene amplification. Although the nature of the amplified sequences needs to be clarified, this experiment demonstrates the potential of reverse chromosome painting with genomic tumor DNA for rapidly mapping the normal chromosomal localization of the DNA from which the amplified sequences were derived. In addition, a weaker staining of chromosomes 10 and X was consistently observed indicating that these chromosomes were present in only one copy in the GBM genome. This rapid approach can be used to analyze cases where no metaphase spreads from the tumor material are available. It does not require any preknowledge of amplified sequences and can be applied to screen large numbers of tumors.


Influence of Coating Application Methods on the Postharvest Quality of Cassava.
Various modes of edible coating application vary in their coat dispersion and film formation, hence the need to determine the most effective mode of application for cassava. Edible surface coatings have been found to be effective in preserving the quality of various food products. However, there are variations in effectiveness among the different coating solutions, hence the need for optimization of the concentrations of the gums used. This study aimed at determining the most efficient coating application method on the cassava postharvest quality. Physiologically mature cassava (variety KME 1) was harvested and divided into seven portions. The various portions were coated using 1.5% xanthan gum, 1.5% xanthan/guar gum, and 2% xanthan/guar gum by both dipping and spraying method. There was no significant difference on the colour, total cyanide, ethylene production, and total phenolic content between the two application methods. The 2% xanthan/guar gum coating showed a significant difference on the dry matter content while the 1.5% xanthan gum coating had a significant difference on the respiration rate and weight loss. The 1.5 xanthan treated roots had a final dry matter content of 72.5% for the sprayed samples and 75.98% for the dipped sample while the 2% xanthan/guar gum treated roots had a final dry matter content of 64.6% and 74.1% for the dipped and sprayed root samples, respectively. The 1.5% xanthan and 2% xanthan/guar gum treated roots showed no significant difference in their action on dry matter content. The 1.5% xanthan/guar dipped and sprayed samples differed significantly on their effect on flesh firmness with final values of 35.4N and 46.1N, respectively, at 20 days after harvest. This study suggested that based on the coating solution and the parameters being observed, there generally was no varying effect of dipping and spraying methods of coating application. The choice of the efficient mode of application to use will depend on other factors such as the easiness of application.


Household survey of HIV incidence in Rwanda: a national observational cohort study.
In Rwanda, HIV prevalence among adults aged 15-49 years has been stable at 3% since 2005. The aim of this study was to characterise HIV incidence across Rwanda. We did a nationally representative, prospective HIV incidence survey for the period of 2013-14, which used two-stage sampling. We randomly selected 492 villages in the first sampling stage and 14 households per village in the second stage. Participants completed a questionnaire and 14 140 people were tested for HIV. 13 728 participants were HIV negative, and were enrolled in the incidence cohort. Participants were retested and surveyed again after 12 months. Weights were calculated as the inverse of the probability to select the villages and the households. The study period was from Nov 5, 2013, to Nov 15, 2014. Among 14 222 respondents from 6792 households, 14 140 were tested for HIV and 13 728 were HIV negative. Of 12 593 people who participated in the endpoint data collection activities, 5965 (47·4%) were men and the mean age was 30 years (SD 10·8). 11 237 (89·2%) participants lived in rural areas, 4826 (38·3%) were single, and 7140 (56·7%) were married or cohabitating. During the year, 35 participants had seroconversion, including 13 men and 22 women, resulting in an overall incidence of 0·27 per 100 person-years (95% CI 0·18-0·35). Incidence was 0·21 per 100 person-years (0·10-0·32) in men and 0·32 per 100 person-years (0·19-0·45) in women. Our findings suggested multiple breakouts, with multiple seroconversions occurring in three villages and two households. Incidence was higher in adults aged 36-45 years (0·37 per 100 person-years, 0·12-0·62; adjusted hazard ratio [aHR] 4·49, 95% CI 1·30-14·70) relative to those aged 16-25, higher in western province (0·57 per 100 person-years, 0·31-0·87; aHR 5·90, 1·33-25·28) relative to the northern province, and higher in urban areas (0·65 per 100 person-years, 0·23-1·07; aHR 3·10, 1·28-6·99) than in rural areas. The incidence of HIV in Rwanda was higher than that previously estimated from models, with outbreaks seeming to contribute to the ongoing epidemic. Characterisation of incident infections can help the national HIV programmes to plan for preventive interventions tailored to the most at risk populations. Global Fund to Fight HIV, Tuberculosis and Malaria, WHO Rwanda, UNAIDS Rwanda, and the Government of Rwanda.


Exercise-based rehabilitation for heart failure.
Previous systematic reviews and meta-analyses consistently show the positive effect of exercise-based rehabilitation for heart failure (HF) on exercise capacity; however, the direction and magnitude of effects on health-related quality of life, mortality and hospital admissions in HF remain less certain. This is an update of a Cochrane systematic review previously published in 2010. To determine the effectiveness of exercise-based rehabilitation on the mortality, hospitalisation admissions, morbidity and health-related quality of life for people with HF. Review inclusion criteria were extended to consider not only HF due to reduced ejection fraction (HFREF or 'systolic HF') but also HF due to preserved ejection fraction (HFPEF or 'diastolic HF'). We updated searches from the previous Cochrane review. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (Issue1, 2013) from January 2008 to January 2013. We also searched MEDLINE (Ovid), EMBASE (Ovid), CINAHL (EBSCO) and PsycINFO (Ovid) (January 2008 to January 2013). We handsearched Web of Science, bibliographies of systematic reviews and trial registers (Controlled-trials.com and Clinicaltrials.gov). Randomised controlled trials of exercise-based interventions with six months' follow-up or longer compared with a no exercise control that could include usual medical care. The study population comprised adults over 18 years and were broadened to include individuals with HFPEF in addition to HFREF. Two review authors independently screened all identified references and rejected those that were clearly ineligible. We obtained full-text papers of potentially relevant trials. One review author independently extracted data from the included trials and assessed their risk of bias; a second review author checked data. We included 33 trials with 4740 people with HF predominantly with HFREF and New York Heart Association classes II and III. This latest update identified a further 14 trials. The overall risk of bias of included trials was moderate. There was no difference in pooled mortality between exercise-based rehabilitation versus no exercise control in trials with up to one-year follow-up (25 trials, 1871 participants: risk ratio (RR) 0.93; 95% confidence interval (CI) 0.69 to 1.27, fixed-effect analysis). However, there was trend towards a reduction in mortality with exercise in trials with more than one year of follow-up (6 trials, 2845 participants: RR 0.88; 95% CI 0.75 to 1.02, fixed-effect analysis). Compared with control, exercise training reduced the rate of overall (15 trials, 1328 participants: RR 0.75; 95% CI 0.62 to 0.92, fixed-effect analysis) and HF specific hospitalisation (12 trials, 1036 participants: RR 0.61; 95% CI 0.46 to 0.80, fixed-effect analysis). Exercise also resulted in a clinically important improvement superior in the Minnesota Living with Heart Failure questionnaire (13 trials, 1270 participants: mean difference: -5.8 points; 95% CI -9.2 to -2.4, random-effects analysis) - a disease specific health-related quality of life measure. However, levels of statistical heterogeneity across studies in this outcome were substantial. Univariate meta-regression analysis showed that these benefits were independent of the participant's age, gender, degree of left ventricular dysfunction, type of cardiac rehabilitation (exercise only vs. comprehensive rehabilitation), mean dose of exercise intervention, length of follow-up, overall risk of bias and trial publication date. Within these included studies, a small body of evidence supported exercise-based rehabilitation for HFPEF (three trials, undefined participant number) and when exclusively delivered in a home-based setting (5 trials, 521 participants). One study reported an additional mean healthcare cost in the training group compared with control of USD3227/person. Two studies indicated exercise-based rehabilitation to be a potentially cost-effective use of resources in terms of gain in quality-adjusted life years (QALYs) and life-years saved. This updated Cochrane review supports the conclusions of the previous version of this review that, compared with no exercise control, exercise-based rehabilitation does not increase or decrease the risk of all-cause mortality in the short term (up to 12-months' follow-up) but reduces the risk of hospital admissions and confers important improvements in health-related quality of life. This update provides further evidence that exercise training may reduce mortality in the longer term and that the benefits of exercise training on appear to be consistent across participant characteristics including age, gender and HF severity. Further randomised controlled trials are needed to confirm the small body of evidence seen in this review for the benefit of exercise in HFPEF and when exercise rehabilitation is exclusively delivered in a home-based setting.


Histogram analysis of renal arterial spin labeling perfusion data reveals differences between volunteers and patients with mild chronic kidney disease.
The spatial heterogeneity of renal perfusion data was analyzed with arterial spin labeling (ASL) data sets in a cohort of subjects with moderately impaired kidney function (ie, glomerular filtration rate >30 mL/min/1.73 m) versus a cohort of healthy volunteers. The potential diagnostic value of a detailed histogram analysis of such perfusion data for detection of mild renal dysfunction was investigated. Eight healthy volunteers and 9 patients with mild renal dysfunction (chronic kidney disease stages 1-3) were included in the study. All subjects underwent ASL perfusion measurements with a 1.5-T magnetic resonance scanner using a flow-sensitive alternating inversion recovery labeling scheme with true fast imaging in steady-state precession data readout. Quantitative perfusion maps were generated using extended Bloch equations. Histogram analysis was performed to quantify the metrics of the perfusion of the renal cortex and the entire parenchyma, respectively. Mean perfusion value (μ), SD of the mean value (σ), peak height (PH), peak position (PP), skewness (s), and kurtosis (k) were computed to describe the distribution of the perfusion values. A significant difference was found in the mean perfusion values computed for the cortex and the parenchyma between healthy volunteers (cortex, 329 ± 53 mL/100 g/min; parenchyma, 301 ± 51 mL/100 g/min) and patients (cortex, 263 ± 81 mL/100 g/min; parenchyma, 244 ± 77 mL/100 g/min). The histogram analysis of the cortical perfusion values also showed a significant difference (P < 0.05) in the main histogram measures between healthy volunteers (PP = 368 ± 65 mL/100 g/min; s = -0.543 ± 0.298; k = 0.371 ± 0.590) and patients (PP = 237 ± 115 mL/100 g/min; s = -0.125 ± 0.581; k = -0.151 ± 0.561). Moderate renal dysfunction is associated with a significant change in the distribution of cortical perfusion values and a reduction of blood perfusion for both the parenchyma and the cortex. The preliminary results reported in this study suggest the importance of a regional assessment of renal perfusion. Histogram analysis of ASL data may help to detect chronic kidney disorders and to monitor their progression in a clinical setting.


Synthesis of the stationary-phase sigma factor sigma s is positively regulated by ppGpp.
Strains of Escherichia coli which lack detectable guanosine 3',5'-bispyrophosphate (ppGpp) display a pleiotropic phenotype that in some respects resembles that of rpoS (katF) mutants. This led us to examine whether ppGpp is a positive regulator of sigma s synthesis. sigma s is a stationary-phase-specific sigma factor that is encoded by the rpoS gene. We found that a ppGpp-deficient strain is defective in sigma s synthesis as cells enter stationary phase in a rich medium, as judged by immunoblots. Under more-defined conditions we found that the stimulation of sigma s synthesis following glucose, phosphate, or amino acid starvation of wild-type strains is greatly reduced in a strain lacking ppGpp. The failure of ppGpp-deficient strains to synthesize sigma s in response to these starvation regimens could indicate a general defect in gene expression rather than a specific dependence of rpoS expression on ppGpp. We therefore tested the effect of artificially elevated ppGpp levels on sigma s synthesis either with mutations that impair ppGpp decay or by gratuitously inducing ppGpp synthesis with a Ptac::relA fusion. In both instances, we observed enhanced sigma s synthesis. Apparently, ppGpp can activate sigma s synthesis under conditions of nutrient sufficiency as well as during entry into stationary phase. This finding suggests that changes in ppGpp levels function both as a signal of imminent stationary phase and as a signal of perturbations in steady-state growth.


Coupling News Sentiment with Web Browsing Data Improves Prediction of Intra-Day Price Dynamics.
The new digital revolution of big data is deeply changing our capability of understanding society and forecasting the outcome of many social and economic systems. Unfortunately, information can be very heterogeneous in the importance, relevance, and surprise it conveys, affecting severely the predictive power of semantic and statistical methods. Here we show that the aggregation of web users' behavior can be elicited to overcome this problem in a hard to predict complex system, namely the financial market. Specifically, our in-sample analysis shows that the combined use of sentiment analysis of news and browsing activity of users of Yahoo! Finance greatly helps forecasting intra-day and daily price changes of a set of 100 highly capitalized US stocks traded in the period 2012-2013. Sentiment analysis or browsing activity when taken alone have very small or no predictive power. Conversely, when considering a news signal where in a given time interval we compute the average sentiment of the clicked news, weighted by the number of clicks, we show that for nearly 50% of the companies such signal Granger-causes hourly price returns. Our result indicates a "wisdom-of-the-crowd" effect that allows to exploit users' activity to identify and weigh properly the relevant and surprising news, enhancing considerably the forecasting power of the news sentiment.


Proliferation, differentiation, and protein synthesis of human osteoblast-like cells (MG63) cultured on previously used titanium surfaces.
This study compared osteoblasts proliferation, differentiation, and protein synthesis on new and used titanium (Ti) disks to test the hypothesis that cleaning and resterilization of previously used Ti disks does not alter cell response to a particular surface. Ti disks of varying roughness were prepared by one of five different treatment regimens. Standard tissue culture plastic was used as a control. Human osteoblast-like cells (MG63) were cultured on the Ti disks and cell proliferation, cell differentiation, RNA synthesis and matrix production (collagen and noncollagen protein; proteoglycans) measured. After their first use, the disks were cleaned, resterilized by autoclaving, and MG63 cells cultured on them as before. At confluence, the same parameters were measured and cell behavior on new and used disks compared. When confluent cultures of cells on plastic were compared to those cultured on new Ti surfaces, cell number was reduced on the roughest surfaces and equivalent to plastic on the other surfaces. Cell number was further reduced when disks with the roughest surfaces were re-used; no differences in cell number could be discerned after cleaning and resterilization. Cell proliferation was inversely related to surface roughness and was less than seen on tissue culture plastic. Re-use of the Ti disks resulted in no change in cell proliferation rate. Alkaline phosphatase specific activity in isolated cells was lowest on the rougher surfaces; no differences between new and used disks were observed. Similarly, enzyme activity in the cell layer was decreased in cultures grown on rougher surfaces, with no effect of prior disk use being noted. RNA synthesis was decreased with respect to plastic in cultures on smoother surfaces and increased on rougher surfaces; prior disk use did not alter RNA synthesis. Collagen production by the cells was decreased on smoother surfaces, but was comparable to tissue culture plastic when grown on rougher surfaces. Non-collagen protein production was unaffected by culture surface and whether or not the disk had been previously used. Proteoglycan synthesis by cells was decreased on all surfaces studied and comparable on both new and used disks. The results of this study indicate that Ti implant surfaces are unaffected by cleaning and resterilization, although rougher surfaces may require more extensive cleaning than smoother ones. This suggests the possibility that implants, in the same patient, could be safely reused. In vivo studies in animals, however, need to be performed before clinical application can be considered.


Identification of myeloperoxidase, alpha-defensin and calgranulin in calcium oxalate renal stones.
In order to understand the mechanism of stone genesis, it is essential to determine the characteristics of macromolecules constituting the urinary stones. We characterized proteins from the inner core and outer matrix of calcium oxalate (CaOx) renal stones. Inner core and outer matrix of CaOx renal stones were separated and proteins were extracted with a buffer containing SDS and beta-mercaptoethanol. Proteins were analyzed and purified by SDS-PAGE and RP-HPLC respectively. The protein bands from gel and protein fractions were sequenced by MALDI TOF mass spectrometry. ELISA, western and slot blot immunoassays were performed to confirm the identity of the proteins in stones and urine of the stone formers. The potential of the identified protein as an effective promoter or inhibitor was assessed by observing their effects on CaOx crystallization using aggregometer. The inner core extract predominantly exhibited protein species in the molecular weight range of 12-14 kDa. However, a 66 kDa band, identified as osteopontin was also detected in the inner core along with outer matrix and in the urine of stone formers and non stone formers. Purification of low molecular weight proteins was carried out by reversed phase HPLC. Tandem mass spectrometry analysis identified them as myeloperoxidase chain A (MPO-A), alpha-defensin, and calgranulin. ELISA, western blot and slot-blot immuno-assays further confirmed their presence restricted to the inner core and not in the outer matrix. Turbidity assays showed that low molecular weight renal stone proteins promoted the aggregation of CaOx crystals. Persistent hyperoxaluria leads to tubular epithelial injury, resulting in the release of these anti-inflammatory proteins. These proteins could have been first adsorbed on CaOx crystals thereby become a part of nucleation process leading to inner matrix formation.


A laboratory-based study to explore the use of honey-impregnated cards to detect chikungunya virus in mosquito saliva.
Mosquito control is implemented when arboviruses are detected in patients or in field-collected mosquitoes. However, mass screening of mosquitoes is usually laborious and expensive, requiring specialized expertise and equipment. Detection of virus in mosquito saliva using honey-impregnated filter papers seems to be a promising method as it is non-destructive and allows monitoring the viral excretion dynamics over time from the same mosquito. Here we test the use of filter papers to detect chikungunya virus in mosquito saliva in laboratory conditions, before proposing this method in large-scale mosquito surveillance programs. We found that 0.9 cm2 cards impregnated with a 50% honey solution could replace the forced salivation technique as they offered a viral RNA detection until 7 days after oral infection of Aedes aegypti and Aedes albopictus mosquitoes with CHIKV.


Retinal function following transpupillary thermotherapy for occult choroidal neovascularization in age-related macular degeneration: a short-term study by focal electroretinography.
To assess short-term changes in macular function after transpupillary thermotherapy (TTT) in patients with occult subfoveal choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD), using focal electroretinography (FERG). Twenty-five patients with occult subfoveal CNV due to AMD were treated with TTT delivered using an infrared (810 nm) diode laser (spot size 3.0 mm, laser power 400-600 mW, duration 60 seconds). All patients were clinically evaluated before, 1 and 6 weeks after treatment. Snellen visual acuity (VA) was measured at each visit. Fluorescein angiography (FA) was performed at baseline and 6 weeks after TTT. Focal ERGs were recorded in all patients immediately before and 1 week after TTT in response to an 18-degree diameter, 41 Hz flickering spot (630 nm) centred on the fovea, presented on a steady background in Maxwellian view. A subgroup of 12 patients was also re-tested by FERG at 6-weeks post-TTT. No significant changes in mean FERG amplitude and phase were observed across the different recording sessions before and after TTT. One week after TTT, four patients had significant (> 2 SD from baseline variability) increases in FERG amplitude and/or phase advances, one had a decrease in amplitude and four had phase delays, compared to baseline. The remaining 15 patients had stable FERGs. Six weeks after TTT, four patients had significant increases in FERG amplitude and/or phase advances, four had decreases in amplitude and/or phase delays, and four had stable FERGs, compared to baseline. Improvement in FERG parameters after TTT was always associated with an improvement in VA and a decrease in exudation. Patients with post-TTT FERG deterioration had stable or deteriorated clinical pictures. At either 1 or 6 weeks post-TTT, the FERG amplitude increase was inversely correlated (p < 0.05) with the baseline FERG amplitude and VA. Three major conclusions can be drawn: in a short-term follow-up, TTT was not found to be associated with significant changes in macular function; FERG improvement was associated with VA improvement, and the increase in FERG amplitude was greatest in patients with the worst baseline acuity.


Fascial components of the myofascial pain syndrome.
Myofascial pain syndrome (MPS) is described as the muscle, sensory, motor, and autonomic nervous system symptoms caused by stimulation of myofascial trigger points (MTP). The participation of fascia in this syndrome has often been neglected. Several manual and physical approaches have been proposed to improve myofascial function after traumatic injuries, but the processes that induce pathological modifications of myofascial tissue after trauma remain unclear. Alterations in collagen fiber composition, in fibroblasts or in extracellular matrix composition have been postulated. We summarize here recent developments in the biology of fascia, and in particular, its associated hyaluronan (HA)-rich matrix that address the issue of MPS.


Using peer education to improve diabetes management and outcomes in a low-income setting: a randomized controlled trial.
Diabetes is an important health burden in Indonesia. However, diabetes management and treatment remain poor, with most people with diabetes in Indonesia not achieving the recommended blood glucose levels. Peer education may have particular potential in low-income settings in complementing diabetes care without being a large additional strain on the health system. This cluster randomized controlled trial aims to identify the effect of the implementation of peer education for patients with type 2 diabetes on diabetes-related outcomes in Aceh, Indonesia, which will complement the diabetes treatment provided at primary-care health posts (puskesmas). Altogether, 29 puskesmas were recruited in Banda Aceh and Aceh Besar, each of which was randomly assigned to either the control or the intervention group. Then, 534 people with diabetes were identified and recruited through their respective puskesmas. The intervention consists of up to two peer education groups per puskesmas, which are led by previously trained people with diabetes. Peer education sessions are held every month for 18 months, with follow-up data being collected 9 and 18 months after the first peer education session. The main objective is to improve diabetes management and the health behavior of participants receiving peer education to reduce their average blood glucose levels as measured by glycated hemoglobin (HbA1c) levels. Secondary outcomes are the effects of peer education on lipid levels, waist circumference, blood pressure, quality of life, treatment adherence, diabetes knowledge, physical activity, and dietary diversity. Data sources for the measurement of outcomes include patient and health facility surveys and biomarker measurements. An economic evaluation will be conducted to assess the cost-effectiveness of the intervention. This trial will contribute to the evidence on the effectiveness and cost-effectiveness of peer education in improving diabetes management in a low-income setting in Indonesia and in other comparable contexts. ISRCTN registry, ISRCTN68253014 . Registered on 18 February 2019.


Puberty and menstruation knowledge among young adolescents in low- and middle-income countries: a scoping review.
This study presents a scoping review of evidence relating to knowledge and experiences of puberty and menstruation among females aged 10-14 years in low- and middle-income countries. Forty-four items from 12 countries were identified from a systematic scoping review and screening of 8083 items. Included studies were quality assessed. A majority (40/44) of studies used school-based samples, and fifteen studies reported on interventions. Girls had inadequate knowledge about menstruation; menarche as a trigger for girls learning about menstruation was common. Adolescents struggled with menstrual hygiene. Negative emotions were associated with menarche and menstrual management. A minority of studies dealt explicitly with puberty. Most girls obtained information about menstruation and/or puberty from their mothers, although mothers were not necessarily girls' preferred source for learning about these topics. Young adolescent girls are under-prepared for puberty and menstruation. Predominantly school-based studies mean we know little about young out-of-school adolescents. The evidence base lags behind the rise in interest from practitioners as well as the development (and evaluation) of puberty and/or menstruation interventions.


Differences in DNA fragmentation following transient cerebral or decapitation ischemia in rats.
The time course of appearance of cells with DNA damage was studied in rats following transient severe forebrain ischemia. This DNA damage could be detected by in situ end-labeling on brain sections. The breaks in DNA appeared selectively by day 1 in the striatum and later in the CA1 region of the hippocampus. It was possible by double labeling to show that there was no DNA damage in astrocytes. The DNA breaks consisted of laddered DNA fragments indicative of an ordered apoptotic type of internucleosomal cleavage, which persisted without smearing for up to 7 days of reperfusion. In contrast, the DNA breaks following ischemia induced by decapitation were random and, after gel electrophoresis, consisted of smeared fragments of multiple sizes. There was some early regional cellular death, restricted to the dentate of the hippocampus, prior to the pannecrotic degeneration. It is concluded that transient forebrain ischemia leads to a type of neuronal destruction that is not random necrosis but that shares some component of the apoptotic cell death pathway.


Biology and genome of Trichinella spiralis.
Clade I nematode species in the genus Trichinella can cause infections in humans that lead to mortality and serious morbidity. There are currently eight recognized species or genotypes that comprise this genus. The species display diverse biological characteristics, the evolutionary significance of which recently has been extensively clarified. Some of that diversity translates into variable importance as zoonotic pathogens, with T. spiralis having the highest significance. Trichinellosis has re-emerged as an important zoonotic infection in various parts of the world, reminding us that control of this infection depends on persistent vigilance. Trichinella species display unique and biologically interesting complexity in interactions with host cells that they inhabit. Significant progress has been made toward understanding details of these interactions. Progress on transcriptomics, proteomics and now genomics offers exciting prospects for accelerating advances in future research. An overview of these parasites regarding biology, significance as zoonotic pathogens and selected research topics is presented here.


No effect of the farming system (organic/conventional) on the bioavailability of apple (Malus domestica Bork., cultivar Golden Delicious) polyphenols in healthy men: a comparative study.
The organic food sales have been increasing during the recent years. It has been hypothesised that organically grown fruits are healthier based on their higher content of phytochemicals. However, data on the bioavailability of phytochemicals from organically or conventionally produced plant foods are scarce. Two human intervention studies were performed to compare the bioavailability of polyphenols in healthy men after ingestion of apples from different farming systems. The administered apples were grown organically and conventionally under defined conditions and characterised regarding their polyphenol content and antioxidant capacity. No significant differences in the polyphenol content and the antioxidant capacity from the organic and conventional farming system were observed. In the short-term intervention study, six men consumed either organically or conventionally produced apples in a randomized cross-over study. After intake of 1 kg apples, phloretin (C (max) 13 + or - 5 nmol/l, t (max) 1.7 + or - 1.2 h) and coumaric acid (C (max )35 + or - 12 nmol/l, t (max) 3.0 + or - 0.8 h) plasma concentrations increased significantly (P < 0.0001) in both intervention groups, without differences between the two farming systems. In the long-term intervention study, 43 healthy volunteers consumed organically or conventionally produced apples (500 g/day; 4 weeks) or no apples in a double-blind, randomized intervention study. In this study, 24 h after the last dosing regime, the apple intake did not result in increasing polyphenol concentrations in plasma and urine compared to the control group suggesting no accumulation of apple polyphenols or degradation products in humans. Our study suggests that the two farming systems (organic/conventional) do not result in differences in the bioavailability of apple polyphenols.


An Overview of Research Opportunities to Increase the Impact of Nutrition Intervention Research in Early Childhood and Education Care Settings According to the RE-AIM Framework.
Objective: To highlight opportunities for future nutrition intervention research within early childhood and education care (ECEC) settings, with a focus on generating evidence that has applicability to real-world policy and practice. Methods: An overview of opportunities to progress the field was developed by the authors using a collaborative writing approach and informed by recent research in the field. The group developed a list of recommendations aligned with the reach, effectiveness, adoption, implementation and maintenance (RE-AIM) framework. Pairs of authors drafted individual sections of the manuscript, which were then reviewed by a separate pair. The first and senior author consolidated all sections of the manuscript and sought critical input on the draft iterations of the manuscript. Results: Interventions that employ digital platforms (reach) in ECEC settings, as well as research in the family day care setting (effectiveness) were identified as areas of opportunities. Research understanding the determinants of and effective strategies for dissemination (adoption), the implementation of nutrition programs, in addition to de-implementation (implementation) of inappropriate nutrition practices, is warranted. For maintenance, there is a need to better understand sustainability and the sustainment of interventions, in addition to undertaking policy-relevant research. Conclusions: The ECEC setting is prime for innovative and practical nutrition intervention research.


Tactile selective attention and body posture: assessing the multisensory contributions of vision and proprioception.
This study addressed the role of proprioceptive and visual cues to body posture during the deployment of tactile spatial attention. Participants made speeded elevation judgments (up vs. down) to vibrotactile targets presented to the finger or thumb of either hand, while attempting to ignore vibrotactile distractors presented to the opposite hand. The first two experiments established the validity of this paradigm and showed that congruency effects were stronger when the target hand was uncertain (Experiment 1) than when it was certain (Experiment 2). Varying the orientation of the hands revealed that these congruency effects were determined by the position of the target and distractor in external space, and not by the particular skin sites stimulated (Experiment 3). Congruency effects increased as the hands were brought closer together in the dark (Experiment 4), demonstrating the role of proprioceptive input in modulating tactile selective attention. This spatial modulation was also demonstrated when a mirror was used to alter the visually perceived separation between the hands (Experiment 5). These results suggest that tactile, spatially selective attention can operate according to an abstract spatial frame of reference, which is significantly modulated by multisensory contributions from both proprioception and vision.


Generalized tube model of biased reptation for gel electrophoresis of DNA.
A theoretical analysis of the reptational motion of DNA in a gel that includes the effects of molecular fluctuations has been used to explain the main features found in experiments involving periodic inversion of the electric field. The resonance-like decrease of the electrophoretic mobility as a function of pulse duration is related to transient "undershoots" in the orientation of the molecule, in agreement with recent experimental data. These features arise from a delicate interplay of internal and center of mass motion of the molecules under pulsed field conditions, and are important for the separation of DNA molecules in the size range 0.2 to 10 million base pairs.


[Solution focused brief therapy in psychiatric practice].
Solution focused brief therapy (SFBT) is a form of psychotherapy that reinforces clients' strengths and focuses on what clients want to replace their problems with. Research shows that SFBT is beneficial and cost-effective and makes the psychiatrist's work more satisfying. It can be used as primary psychotherapy, for instance during crisis intervention, or even as an addition to pharmacotherapy.


Pulmonary response to intratracheal instillation of ultrafine versus fine titanium dioxide: role of particle surface area.
The production and use of nanoparticles is growing rapidly due to the unique physical and chemical properties associated with their nano size and large surface area. Since nanoparticles have unique physicochemical properties, their bioactivity upon exposure to workers or consumers is of interest. In this study, the issue of what dose metric (mass dose versus surface area dose) is appropriate for toxicological studies has been addressed. Rats were exposed by intratracheal instillation to various doses of ultrafine or fine TiO2. At 1, 7, or 42 days post-exposure, inflammatory and cytotoxic potential of each particle type was compared on both a mass dosage (mg/rat) as well as an equal surface area dosage (cm2 of particles per cm2 of alveolar epithelium) basis. The findings of the study show that on a mass basis the ultrafine particles caused significantly more inflammation and were significantly more cytotoxic than the fine sized particles. However, when doses were equalized based on surface area of particles delivered, the ultrafine particles were only slightly more inflammogenic and cytotoxic when compared to the fine sized particles. Lung burden data indicate that ultrafine TiO2 appears to migrate to the interstitium to a much greater extent than fine TiO2. This study suggests that surface area of particles may be a more appropriate dose metric for pulmonary toxicity studies than mass of particles.


Electrically stimulated acupuncture increases renal blood flow through exosome-carried miR-181.
Acupuncture with low-frequency electrical stimulation (Acu/LFES) can prevent muscle atrophy by increasing muscle protein anabolism in mouse models of chronic kidney disease. During the treatment of muscle wasting, we found that Acu/LFES on the gastrocnemius muscle of the leg enhances renal blood flow. We also found that Acu/LFES increases exosome abundance and alters exosome-associated microRNA expression in the circulation. When exosome secretion was blocked using GW4869, the Acu/LFES-induced increase in renal blood flow was limited. This provided evidence that the increased renal blood flow is exosome mediated. To identify how exosomes regulate renal blood flow, we performed microRNA deep sequencing in exosomes isolated from treated and untreated mouse serum and found that the 34 microRNAs are altered by Acu/LFES. In particular, miR-181d-5p is increased in the serum exosome of Acu/LFES-treated mice. In silico searching suggested that miR-181d-5p could target angiotensinogen. Using a luciferase reporter assay, we demonstrated that miR-181 directly inhibits angiotensinogen. When Acu/LFES-treated muscle was excised and incubated in culture medium, we found that the amount of exosomes and miR-181d-5p was increased in the medium providing evidence that Acu/LFES can increase miR-181 secretion. We conclude that Acu/LFES on leg hindlimb increases miR-181 in serum exosome leading to increased renal blood flow. This study provides important new insights about the mechanism(s) by which acupuncture may regulation of muscle-organ cross talk through exosome-derived microRNA.


The expression of TIA-1+ cytolytic-type granules and other cytolytic lymphocyte-associated markers in CD30+ anaplastic large cell lymphomas (ALCL): correlation with morphology, immunophenotype, ultrastructure, and clinical features.
Anaplastic large cell lymphomas (ALCL) are a heterogeneous group of CD30+ large cell lymphomas; the most characteristic type have a T or null cell phenotype, often express epithelial membrane antigen (EMA) and cytolytic lymphocyte markers, and often possess a nonrandom t(2;5)(p23;q35) chromosomal translocation. We studied 22 (19 T, 1 null, 2 B cell) ALCL, including four primary cutaneous ALCL (PC-ALCL), for the expression of TIA-1, the cytotoxic T lymphocyte (CTL) or natural killer (NK) cell-associated antigens CD4, CD8, betaF1, TCRdelta1, CD56, and CD57, the ALCL-associated antigens p80 and EMA, and the Hodgkin's disease-associated marker CD15 to better define the relationship of these markers to histological subtype, primary site, and patient clinical characteristics. TIA-1 expression was seen in 12 of 20 (60%) T or null cell ALCLs with a cytoplasmic, granular distribution. Ultrastructural studies showed cytotoxic-type granules (dense core, multivesicular, and intermediate types) with TIA-1 localized to granules on immunogold labeling. TIA-1 staining strongly correlated with young patient age (< or = 32 years, P < .05) and EMA expression (P < .05). Excluding the four PC-ALCL cases, TIA-1 staining also correlated with p80 expression (P < .05) in all of the T cell cases. Three CD15+ cases were TIA-1-. TIA-1 expression in T or null cell ALCL was seen in all morphological subtypes (2 of 2 small cell variant, 3 of 4 monomorphic variant, and 7 of 14 pleomorphic variant) and primary tumor sites (6 of 14 nodal, 2 of 4 primary cutaneous, 2 of 2 bone, and 2 of 2 soft tissue). TIA-1+ granules were seen in all subsets: 5 of 6 CD4+, 1 of 2 CD8+, 4 of 8 CD56+, and 1 of 2 CD57+ ALCL. Of note, 4 of 10 T or null cell ALCL expressed gammadelta T-cell receptors (TCR), whereas only 1 of 10 T or null cell ALCL was alphabeta TCR+; TCR were not detected in five cases. TIA-1 was expressed by 3 of 4 gammadelta TCR+ ALCL and 1 of 1 alphabeta TCR+ ALCL. These data support a cytotoxic lymphocyte phenotype in most T or null cell ALCL and suggest that some T cell ALCL are derived from cytolytic CD4+ T cells, gammadelta T cells, or NK-like (CD56+ or CD57+) T cells.


Tobramycin and bicarbonate synergise to kill planktonic Pseudomonas aeruginosa, but antagonise to promote biofilm survival.
Increasing antibiotic resistance and the declining rate at which new antibiotics come into use create a need to increase the efficacy of existing antibiotics. The aminoglycoside tobramycin is standard-of-care for many types of Pseudomonas aeruginosa infections, including those in the lungs of cystic fibrosis (CF) patients. P. aeruginosa is a nosocomial and opportunistic pathogen that, in planktonic form, causes acute infections and, in biofilm form, causes chronic infections. Inhaled bicarbonate has recently been proposed as a therapy to improve antimicrobial properties of the CF airway surface liquid and viscosity of CF mucus. Here we measure the effect of combining tobramycin and bicarbonate against P. aeruginosa, both lab strains and CF clinical isolates. Bicarbonate synergises with tobramycin to enhance killing of planktonic bacteria. In contrast, bicarbonate antagonises with tobramycin to promote better biofilm growth. This suggests caution when evaluating bicarbonate as a therapy for CF lungs infected with P. aeruginosa biofilms. We analyse tobramycin and bicarbonate interactions using an interpolated surface methodology to measure the dose-response function. These surfaces allow more accurate estimation of combinations yielding synergy and antagonism than do standard isobolograms. By incorporating predictions based on Loewe additivity theory, we can consolidate information on a wide range of combinations that produce a complex dose-response surface, into a single number that measures the net effect. This tool thus allows rapid initial estimation of the potential benefit or harm of a therapeutic combination. Software code is freely made available as a resource for the community.


Evidence that GABA rho subunits contribute to functional ionotropic GABA receptors in mouse cerebellar Purkinje cells.
Ionotropic gamma-amino butyric acid (GABA) receptors composed of heterogeneous molecular subunits are major mediators of inhibitory responses in the adult CNS. Here, we describe a novel ionotropic GABA receptor in mouse cerebellar Purkinje cells (PCs) using agents reported to have increased affinity for rho subunit-containing GABA(C) over other GABA receptors. Exogenous application of the GABA(C)-preferring agonist cis-4-aminocrotonic acid (CACA) evoked whole-cell currents in PCs, whilst equimolar concentrations of GABA evoked larger currents. CACA-evoked currents had a greater sensitivity to the selective GABA(C) antagonist (1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid (TPMPA) than GABA-evoked currents. Focal application of agonists produced a differential response profile; CACA-evoked currents displayed a much more pronounced attenuation with increasing distance from the PC soma, displayed a slower time-to-peak and exhibited less desensitization than GABA-evoked currents. However, CACA-evoked currents were also completely blocked by bicuculline, a selective agent for GABA(A) receptors. Thus, we describe a population of ionotropic GABA receptors with a mixed GABA(A)/GABA(C) pharmacology. TPMPA reduced inhibitory synaptic transmission at interneurone-Purkinje cell (IN-PC) synapses, causing clear reductions in miniature inhibitory postsynaptic current (mIPSC) amplitude and frequency. Combined application of NO-711 (a selective GABA transporter subtype 1 (GAT-1) antagonist) and SNAP-5114 (a GAT-(2)/3/4 antagonist) induced a tonic GABA conductance in PCs; however, TPMPA had no effect on this current. Immunohistochemical studies suggest that rho subunits are expressed predominantly in PC soma and proximal dendritic compartments with a lower level of expression in more distal dendrites; this selective immunoreactivity contrasted with a more uniform distribution of GABA(A) alpha1 subunits in PCs. Finally, co-immunoprecipitation studies suggest that rho subunits can form complexes with GABA(A) receptor alpha1 subunits in the cerebellar cortex. Overall, these data suggest that rho subunits contribute to functional ionotropic receptors that mediate a component of phasic inhibitory GABAergic transmission at IN-PC synapses in the cerebellum.


Explosive type of moderate-resistance training induces functional, cardiovascular, and molecular adaptations in the elderly.
Current recommendations aimed at reducing neuromuscular and functional loss in aged muscle have identified muscle power as a key target for intervention trials, although little is known about the biological and cardiovascular systemic response in the elderly. This study investigated the effects of 12 weeks of low-frequency, moderate-intensity, explosive-type resistance training (EMRT) on muscle strength and power in old community-dwelling people (70-75 years), monitoring functional performance linked to daily living activities (ADL) and cardiovascular response, as well as biomarkers of muscle damage, cardiovascular risk, and cellular stress response. The present study provides the first evidence that EMRT was highly effective in achieving a significant enhancement in muscular strength and power as well as in functional performance without causing any detrimental modification in cardiovascular, inflammatory, and damage parameters. Moreover, trained elderly subjects showed an adaptive response at both systemic and cellular levels by modulation of antioxidant and stress-induced markers such as myeloperoxidase (MPO), heat shock protein 70 (Hsp70) and 27 (Hsp27), and thioredoxin reductase 1 (TrxR1).


Carcinoid tumor of the cecum presenting with acute appendicitis: a case report.
Acute appendicitis is the most common cause of acute abdomen. Carcinoid tumor of the appendix is a rare incidental finding that can present with the clinical picture of acute appendicitis. During open surgery for acute appendicitis, a 3 cm solid mass, not noticed externally, was palpated at the base of the appendix. The mass and the appendix were excised by en-bloc wedge resection. The histopathological examination of the lesion revealed carcinoid tumor. The aim of this presentation is to remind that neoplasms of the appendix may, although rarely, present the clinical picture of acute appendicitis, and to highlight that they, particularly those located at the base of the appendix and in cecum, may be overlooked during laparoscopy. The importance of preoperative computerized tomography ins uch cases has to be underlined.


Religious affiliation, internalized homophobia, and mental health in lesbians, gay men, and bisexuals.
Most religious environments in the United States do not affirm homosexuality. The authors investigated the relationship between exposure to nonaffirming religious environments and internalized homophobia and mental health in a sample of lesbians, gay men, and bisexuals (LGBs) in New York City. Guided by minority stress theory, the authors hypothesized that exposure to nonaffirming religious settings would lead to higher internalized homophobia, more depressive symptoms, and less psychological well-being. The authors hypothesized that Black and Latino LGBs would be more likely than White LGBs to participate in nonaffirming religious settings and would therefore have higher internalized homophobia than White LGBs. Participants were 355 LGBs recruited through community-based venue sampling and evenly divided among Black, Latino, and White race or ethnic groups and among age groups within each race or ethnic group, as well as between women and men. Results supported the general hypothesis that nonaffirming religion was associated with higher internalized homophobia. There was no main effect of nonaffirming religion on mental health, an unexpected finding discussed in this article. Latinos, but not Blacks, had higher internalized homophobia than Whites, and as predicted, this was mediated by their greater exposure to nonaffirming religion.


Organ donation after medical assistance in dying or cessation of life-sustaining treatment requested by conscious patients: the Canadian context.
In June 2016, following the decision of the Supreme Court of Canada to decriminalise assistance in dying, the Canadian government enacted Bill C-14, legalising medical assistance in dying (MAID). In 2014, the province of Quebec had passed end-of-life care legislation making MAID available as of December 2015. The availability of MAID has many implications, including the possibility of combining this practice with organ donation through the controlled donation after cardiac death (cDCD) protocol. cDCD most often occurs in cases where the patient has a severe neurological injury but does not meet all the criteria for brain death. The donation is subsequent to the decision to withdraw life-sustaining treatment (LST). Cases where patients are conscious prior to the withdrawal of LST are unusual, and have raised doubts as to the acceptability of removing organs from individuals who are not neurologically impaired and who have voluntarily chosen to die. These cases can be compared with likely scenarios in which patients will request both MAID and organ donation. In both instances, patients will be conscious and competent. Organ donation in such contexts raises ethical issues regarding respect for autonomy, societal pressure, conscientious objections and the dead-donor rule. In this article, we look at relevant policies in other countries and examine the ethical issues associated with cDCD in conscious patients who choose to die.


Association between hearing aid use and all-cause and cause-specific dementia: an analysis of the UK Biobank cohort.
Dementia and hearing loss are both highly prevalent conditions among older adults. We aimed to examine the association between hearing aid use and risk of all-cause and cause-specific dementia among middle-aged and older-aged adults, and to explore the roles of mediators and moderators in their association. We used data from the UK Biobank, a population-based cohort study, which recruited adults aged 40-69 years between 2006 and 2010 across 22 centres in England, Scotland, and Wales. We used Cox proportional hazards models to estimate hazard ratios (HRs) and 95% CIs between self-reported hearing aid use status (hearing loss with or without hearing aids) at baseline and risk of dementia (all-cause dementia, Alzheimer's disease, vascular dementia, and non-Alzheimer's disease non-vascular dementia). Dementia diagnoses were ascertained using hospital records and death-register data. We also analysed the roles of mediators (self-reported social isolation, loneliness, and mood) and moderators (self-reported education and income, smoking, morbidity, and measured APOE allele status). After the exclusion of people who did not answer the question on hearing difficulties (n=25 081 [5·0%]) and those with dementia at baseline visit (n=283 [0·1%]), we included 437 704 people in the analyses. Compared with participants without hearing loss, people with hearing loss without hearing aids had an increased risk of all-cause dementia (HR 1·42 [95% CI 1·29-1·56]); we found no increased risk in people with hearing loss with hearing aids (1·04 [0·98-1·10]). The positive association of hearing aid use was observed in all-cause dementia and cause-specific dementia subtypes (Alzheimer's disease, vascular dementia, and non-Alzheimer's disease non-vascular dementia). The attributable risk proportion of dementia for hearing loss was estimated to be 29·6%. Of the total association between hearing aid use and all-cause dementia, 1·5% was mediated by reducing social isolation, 2·3% by reducing loneliness, and 7·1% by reducing depressed mood. In people with hearing loss, hearing aid use is associated with a risk of dementia of a similar level to that of people without hearing loss. With the postulation that up to 8% of dementia cases could be prevented with proper hearing loss management, our findings highlight the urgent need to take measures to address hearing loss to improve cognitive decline. National Natural Science Foundation of China and Shandong Province, Taishan Scholars Project, China Medical Board, and China Postdoctoral Science Foundation.


Expression patterns of Bmi-1 and p16 significantly correlate with overall, disease-specific, and recurrence-free survival in oropharyngeal squamous cell carcinoma.
The objective of this study was to link expression patterns of B-cell-specific Moloney murine leukemia virus integration site 1 (Bmi-1) and p16 to patient outcome (recurrence and survival) in a cohort of 252 patients with oral and oropharyngeal squamous cell cancer (OSCC). Expression levels of Bmi-1 and p16 in samples from 252 patients with OSCC were evaluated immunohistochemically using the tissue microarray method. Staining intensity was determined by calculating an intensity reactivity score (IRS). Staining intensity and the localization of expression within tumor cells (nuclear or cytoplasmic) were correlated with overall, disease-specific, and recurrence-free survival. The majority of cancers were localized in the oropharynx (61.1%). In univariate analysis, patients who had OSCC and strong Bmi-1 expression (IRS >10) had worse outcomes compared with patients who had low and moderate Bmi-1 expression (P = .008; hazard ratio [HR], 1.82; 95% confidence interval [CI], 1.167-2.838); this correlation was also observed for atypical cytoplasmic Bmi-1 expression (P = .001; HR, 2.164; 95% CI, 1.389-3.371) and for negative p16 expression (P < .001; HR, 0.292; 95% CI, 0.178-0.477). The combination of both markers, as anticipated, had an even stronger correlation with overall survival (P < .001; HR, 8.485; 95% CI, 4.237-16.994). Multivariate analysis demonstrated significant results for patients with oropharyngeal cancers, but not for patients with oral cavity tumors: Tumor classification (P = .011; HR, 1.838; 95%CI, 1.146-2.947) and the combined marker expression patterns (P < .001; HR, 6.254; 95% CI, 2.869-13.635) were correlated with overall survival, disease-specific survival (tumor classification: P = .002; HR, 2.807; 95% CI, 1.477-5.334; combined markers: P = .002; HR, 5.386; 95% CI, 1.850-15.679), and the combined markers also were correlated with recurrence-free survival (P = .001; HR, 8.943; 95% CI, 2.562-31.220). Cytoplasmic Bmi-1 expression, an absence of p16 expression, and especially the combination of those 2 predictive markers were correlated negatively with disease-specific and recurrence-free survival in patients with oropharyngeal cancer. Therefore, the current results indicate that these may be applicable as predictive markers in combination with other factors to select patients for more aggressive treatment and follow-up.


Laboratory Medicine in Palestine.
Laboratory Medicine (LM) is one of the cornerstones of healthcare. In Palestine, 3.5% of health expenditure is allocated to clinical laboratories. The Palestinian Ministry of Health (MOH) started to invest in the development and expansion of laboratory services including the introduction of full automation in blood banks and histopathology, molecular testing in microbiology, and testing for autoimmune diseases and metabolic disorders. Improvements have not been limited to new tests but also included external quality assurance (EQA) and accreditation programs. Latest investments have cut the costs of purchasing tests from outside MOH by more than 3.6 million dollars during the last five years. Al-Quds University established a Center for External Quality Control in LM which was supported by the Palestinian MOH and the National Metrology Institute of Germany. This has led to a significant improvement in the performance of the affiliated laboratories. An accreditation unit was established within the MOH, yet the number of laboratories that have been accredited with ISO-15189 are still limited but significantly increasing.The academic institutions have been working in parallel to the MOH in improving LM in Palestine. A new academic curriculum for LM has been developed according to the quality standards of curricula-based competencies determined by the needs of the labor market and the emerging technologies. Despite the invested efforts, still, the Palestinian LM suffers from shortage of human resources which are qualified in the new emerging diagnostic approaches.


Ultraspiracle promotes the nuclear localization of ecdysteroid receptor in mammalian cells.
The heterodimer consisting of ecdysteroid receptor (EcR) and ultraspiracle (USP), both of which are members of the nuclear receptor superfamily, is considered to be the functional ecdysteroid receptor. Here we analyzed the subcellular distribution of EcR and USP fused to fluorescent proteins. The experiments were carried out in mammalian COS-7, CHO-K1 and HeLa cells to facilitate investigation of the subcellular trafficking of EcR and USP in the absence of endogenous expression of these two receptors. The distribution of USP tagged with a yellow fluorescent protein (YFP-USP) was almost exclusively nuclear in all cell types analyzed. The nuclear localization remained constant for at least 1 day after the first visible signs of expression. In contrast, the intracellular distribution of EcR tagged with a yellow fluorescent protein (YFP-EcR) varied and was dependent on time and cell type, although YFP-EcR alone was also able to partially translocate into the nuclear compartment. Coexpression of YFP-EcR with USP tagged with a cyan fluorescent protein (CFP-USP) resulted in exclusively nuclear localization of both proteins in all cell types analyzed. The USP-induced nuclear localization of YFP-EcR was stable for at least 20 hours. These experiments suggest that USP has a profound effect on the subcellular distribution of EcR.


Geographical variation in hospital admission rates: an analysis of workload in the Oxford region, England.
To measure variation in hospital admission rates between health districts in part of the English NHS, comparing a wide range of medical and surgical conditions. Retrospective analysis of interdistrict variation using linked routine hospital admission data. Comparisons were also made with levels of variation reported from the USA. Oxford Regional Health Authority, 1979-86. Six district health authorities--total study population 2.1 million people, 1.6 million hospital admissions. Age and sex standardised hospital admission rates for resident populations for individual operations and diagnoses; systematic components of variation (SCV). Of 118 standard operation groups, 38 (26% of surgical workload) showed high variation (SCV 16 or more) and 40 (36% of surgical workload) showed low variation (SCV < 4). Operations (SCV) with very low levels of variation included prostatectomy (0.1), inguinal herniorraphy (0.9), and cholecystectomy (1.3). Rates were more variable for myringotomy (3.7), hysterectomy (4.3), dilatation and curettage (5.6), and tonsillectomy (6.2). The SCV was high for only four of the 40 commonest medical causes of admission, and was low for 18 of them. Most admissions in the Oxford region were for conditions that did not show a great deal of variation in admission rates. The level of variation for many surgical procedures was less than that reported in studies from the USA. Variation was no greater for medical causes of admission than for surgical conditions. Large scale variation may not be an inevitable consequence of autonomous clinical practice.


A region in the seven-transmembrane domain of the human Ca2+ receptor critical for response to Ca2+.
Of 12 naturally occurring, activating mutations in the seven-transmembrane (7TM) domain of the human Ca2+ receptor (CaR) identified previously in subjects with autosomal dominant hypocalcemia (ADH), five appear at the junction of TM helices 6 and 7 between residue Ile819 and Glu837. After identifying a sixth activating mutation in this region, V836L, in an ADH patient, we studied the remaining residues in this region to determine whether they are potential sites for activating mutations. Alanine-scanning mutagenesis revealed five additional residues in this region that when substituted by alanine led to CaR activation. We also found that, whereas E837A did not activate the receptor, E837D and E837K mutations did. Thus, region Ile819-Glu837 of the 7TM domain represents a "hot spot" for naturally occurring, activating mutations of the receptor, and most of the residues in this region apparently maintain the 7TM domain in its inactive configuration. Unique among the residues in this region, Pro823, which is highly conserved in family 3 of the G protein-coupled receptors, when mutated to either alanine or glycine, despite good expression severely impaired CaR activation by Ca2+. Both the P823A mutation and NPS 2143, a negative allosteric modulator that acts on the 7TM through a critical interaction with Glu837, blocked activation of the CaR by various ADH mutations. These results suggest that the 7TM domain region Ile819-Glu837 plays a key role in CaR activation by Ca2+. The implications of our finding that NPS 2143 corrects the molecular defect of ADH mutations for treatment of this disease are also discussed.


Effect of lipopolysaccharide on the xenobiotic-induced expression and activity of hepatic cytochrome P450 in mice.
Infection-associated inflammation can alter the expression levels and functions of cytochrome P450s (CYPs). Cyp gene expression is regulated by the activation of several nuclear receptors, including pregnane X receptor (PXR), constitutive androstane receptor (CAR), and aryl hydrocarbon receptor (AhR). These receptors can be activated by xenobiotics, including medicines. Here, to study the xenobiotic-induced fluctuations in CYP during inflammation, we examined the effect of lipopolysaccharide (LPS) treatment on the level of mRNAs encoding hepatic CYPs induced by xenobiotic-activated nuclear receptors, in mice. Both the mRNA induction of Cyp genes and the metabolic activities of CYP proteins were examined. LPS treatment caused a significant decrease in the induced expression of the mRNAs for Cyp3a11, 2c29, 2c55, and 1a2, but not for Cyp2b10. To assess the CYP enzymatic activities, CYP3A-mediated testosterone 6β-hydroxylation and the intrinsic clearance (CL(int)) of nifedipine in liver microsomes were measured in mice treated with the xenobiotic pregnenolone-16alpha-carbonitrile (PCN) with or without LPS administration. Both assays revealed that the CYP3A activity, which was induced by PCN, declined significantly after LPS treatment, and this decline correlated with the Cyp3a11 mRNA level. In addition, we found that the mRNAs for interleukin (IL)-1β and tumor necrosis factor (TNF) α were increased after treatment with LPS plus xenobiotics. Our findings demonstrated that LPS treatment reduces the PXR- and AhR-mediated, and possibly CAR-mediated Cyp gene expression and further suggest that these decreases are dependent on inflammatory cytokines in the liver.


Early malfunction of transvenous pacemaker electrodes. A three-center study.
A 3-year study by three medical centers has revealed a 1-year electrode malfunction rate of 7.4%; most malfunctions occurred within the first 30 days. The incidence of unavoidable early malfunction (3.2%) fell within the 5% standards suggested by the committee report of the Inter-Society Committee on Heart Diseases. Incidences of obscure cause (3.2%) may be difficult to identify prospectively and may be, to a certain extent, unavoidable. The majority of the malfunctions (4.2%) showed specific clues that indicated that they were preventable. Successful repositioning was achieved on the first attempt in 80.6% of the cases with malfunction, and only 0.7% required ultimate myocardial electrode implantation. The principal clues to potentially unsatisfactory positioning included the presence of a large right ventricle with or without tricuspid insufficiency, current thresholds greater than 0.5 mA and ST-segment deviations on the intracardiac electrogram of less than 2 mV. Electrode malfunction may be more common with bipolar than with unipolar electrodes; but significant differences in the incidence of malfunction among different unipolar electrodes were observed. These data indicate that further developments in transvenous electrode design are warranted.