Schwannoma (Neurilemmoma) on the Base of the Tongue: A Rare Clinical Case.
Schwannomas are slow-growing benign tumors. They can arise from any peripheral nerve, including the cranial nerves (except the olfactory and optic nerves), spinal nerves, and autonomic nerves. Schwannomas of the head and neck account for 25-40% of all cases. However, intra-oral schwannomas account for only 1% of all head and neck tumors. Complete surgical excision is the treatment of choice. Malignant transformation and recurrence following this treatment are rare. A 20-year-old woman presented with a slow-growing mass over the back of her tongue first noticed 8 months before. Examination of the oral cavity exposed a 4 × 4 cm mass over the posterior aspect of the tongue. The remaining oral cavity examination was normal, with no cervical lymph node enlargement. The patient underwent excisional biopsy by the trans-oral approach under general anesthesia. Histopathological reports discovered features of schwannoma. The patient was followed up for 1 year; she had an uneventful recovery and no evidence of recurrence. We report a case of schwannoma over the base of the tongue, a rare location for this type of tumor. In this article we report a case of schwannoma over the base of the tongue. Despite the rarity of this condition, physicians should consider schwannoma as a differential diagnosis for a mass over the tongue, as there can be a favorable outcome and prognosis for the patient when this condition is correctly identified.


Anti-atherogenic effects of seabuckthorn (Hippophaea rhamnoides) seed oil.
Seabuckthorn (SBT) seed oil is a rich source of unsaturated fatty acids, phytosterols, carotenoids and flavonoids, which are known to have significant anti-atherogenic and cardioprotective activity. The anti-atherogenic activity of supercritical CO(2) extracted SBT seed oil was evaluated in white albino rabbits fed on high cholesterol diet for 60 days. The study was performed on 20 male healthy rabbits divided into four groups of 5 animals each. Group I - control, group II - SBT seed oil, group III - cholesterol (1%) for 60 days, group IV - cholesterol+SBT seed oil. After 30 days of high cholesterol diet, group IV rabbits received 1 ml of SBT seed oil daily for 30 days. Blood total cholesterol (TC), LDL-cholesterol (LDL-C), HDL-cholesterol (HDL-C) and triglyceride (TG) levels were measured before and after the administration of SBT seed oil. The vasorelaxant activity of the seed oil was studied in vitro using aortic ring model technique and changes in isometric force were recorded using a polygraphic recording system. Accumulation of cholesterol in the aorta was studied using Sudan-IV staining technique. SBT seed oil feeding to normal rabbits for 18 days caused a significant decline in plasma cholesterol, LDL-C, atherogenic index (AI) and LDL/HDL ratio. The HDL-C levels, HDL-C/TC ratio (HTR) and vasorelaxant activity of the aorta were significantly increased. In cholesterol-fed animals the TC, TG, LDL-C and AI were significantly increased and showed a decline following seed oil administration. The increase in HDL-C was more marked in seed oil treated hypercholesterolemic animals. The acetylcholine-induced vasorelaxant activity was significantly decreased in cholesterol-fed animals and could be restored to that of normal values by seed oil administration. These observations suggest that supercritical CO(2) extracted SBT seed oil has significant anti-atherogenic and cardioprotective activity.


Effect of atrazine on metamorphosis and sexual differentiation in Xenopus laevis.
There is a growing international concern that commonly used environmental contaminants have the potential to disrupt the development and functioning of the reproductive system in amphibians. One such chemical of interests is the herbicide atrazine. Effects of atrazine on sex differentiation were studied using wild-type Xenopus laevis tadpoles and all-ZZ male cohorts of X. laevis tadpoles, produced by mating wild-type ZZ male to sex-reversed ZZ male (female phenotype). Stage 49 tadpoles were exposed to 0.1-100 ppb atrazine or 0.27 ppb (1 nM) 17beta-estradiol (E(2)) until all larvae completed metamorphosis (stage 66). Metamorphosis, gonadal morphology and histology, CYP19 (P450 aromatase) mRNA induction, and hepatic vitellogenin (VTG) induction were investigated. Effects of atrazine on VTG-induction were also assessed in vitro in primary-cultured X. laevis hepatocytes. Atrazine had no effect on metamorphosis of developing wild-type or all-male X. laevis larvae. Statistical increase in female ratios was observed in 10 and 100 ppb atrazine groups in comparison with control group. While no hermaphroditic froglet was observed in all atrazine groups. In ZZ males, sex reversal was induced by 0.27 ppb E(2), but not by atrazine at concentrations of 0.1 and 1.0 ppb. In addition, neither P450 aromatase mRNA in the gonad nor hepatic VTG were induced by atrazine. Furthermore, VTG was not induced by 1000 ppb atrazine in primary-cultured hepatocytes. Our results indicate that female ratios in developing X. laevis tadpoles were increased by 10 and 100 ppb atrazine under the present experimental conditions. While the other endpoints showed no effect in the range of 0.1-100 ppb atrazine. These results suggest that effect of atrazine on sexual differentiation was not caused by estrogenic action and has no induction ability of P450 aromatase gene in gonad.


Left-right asymmetric expression of BbPtx, a Ptx-related gene, in a lancelet species and the developmental left-sidedness in deuterostomes.
The long-standing question of how asymmetric development or asymmetric body structures in lancelets (amphioxus) are phylogenetically related to the body plan of other animals is still untouched. Three anterior structures, the preoral pit, club-shaped gland and mouth, are remarkable asymmetric features in developing lancelets that all open on the left side of the body. A Ptx-related gene, BbPtx is the first identified transcription factor gene with an asymmetrical expression pattern in lancelets similar to that in vertebrates, and thus it may provide a clue for the above question. Expression of the BbPtx gene is first detected at the dorsal margin of the blastopore in early mid-gastrulae and then becomes restricted to the left anterodorsal wall of the primitive gut and to the developing left somitocoelomic system. Expression continues on the left side in the developing preoral pit, club-shaped gland and mouth as well as in the mesoderm at the caudal end. Unlike D-Ptx1 in Drosophila, BbPtx is not coexpressed with a fork head gene in lancelets; instead the two genes are expressed in a complementary fashion on the left side of the embryo. The expression pattern of BbPtx is not compatible with the calcichordate hypothesis of Jefferies, in which the proposed ancestor of chordates rotated its tail 90 degrees counterclockwise in relation to the head/trunk. The expression of both BbPtx and vertebrate Pitx2 in tissues derived from the coelom implies that the left-right asymmetric development has a common origin between cephalochordates and vertebrates. Considering the development of the coelom in deuterostomes, however, left-right asymmetric development involving Pitx2-related genes is rather likely to be a primitive character shared among deuterostomes.


A randomized controlled trial of direct access for laparoscopic sterilization.
The aim of this study was to establish and evaluate a direct access service for laparoscopic sterilization. A pragmatic randomized controlled trial was carried out on a total of 232 women referred to Aberdeen Royal Infirmary for sterilization between 1 June 1996 and 31 March 1997, from 57 general practices around Aberdeen, Scotland, comprising 75 from general practices that had received criteria for direct access, and 157 from control practices. The main outcome measures were: patients' waiting times to appointments and operation, satisfaction, short-term regret, operative complications and costs; conjoint analysis of patient preferences; GPs' adherence to referral criteria; GP and gynaecologist satisfaction; and NHS costs. Analysis was by intention to treat. There were no inappropriate direct referrals. Waiting time was lower in the intervention group, but there were more visits post-operatively to the GP. Patient and doctor satisfaction was equally high in both groups. There was no difference in operative complication rate, nor in total cost to patients or to the NHS. GPs and gynaecologists strongly supported direct access, but women preferred to meet both a gynaecologist and a GP before sterilization (routine referral). Other attributes of care important to patients included written information, although waiting time was not important. Of all women referred for sterilization during the study period, only 31% were suitable for direct referral. Patients preferred routine referral, and there were no differences in patient costs or satisfaction. There was, however, strong medical support for direct referral. Restrictive direct referral criteria may limit the uptake. These findings are important for future planning of direct referral services. Important methodological lessons were also learnt about the conduct of trials involving the primary-secondary care interface.


Influence of fiber posts on the fracture resistance of endodontically treated premolars with different dental defects.
This study aimed to evaluate the influence of quartz fiber post placement on the fracture resistance of endodontically treated premolars with different dental defects under dynamic loading. Fifty extracted single-rooted mandibular premolars were randomized into five groups. Each group was prepared according to numbers of residual walls ranged from 0 to 4. Then each group was divided into two subgroups with one restored with quartz fiber posts and the other without posts. In no-post groups, gutta percha point 2 mm below cemento-enamel junction was removed. Composite resin was adapted to the well and used to shape the core directly. Each tooth was restored with a complete metal crown. Dynamic loading was carried out in a masticatory simulator with a nominal load of 50 N at 2 Hz for 300 000 loading cycles. Then a quasi-statically load was applied in a universal testing machine 30° to the long axis with a crosshead speed of 1 mm⋅min(-1) until fracture. Data were analyzed with one-way analysis of variance and pairwise comparison (P<0.05). No specimens failed during dynamic loading. The fracture resistance enhanced with the increase of numbers of coronal walls and the differences were significant (P<0.05). Placement of fiber posts had a significant effect when fewer than two walls remained (P<0.05), but it had no significant influence in groups with two, three or four walls (P>0.05). Fiber post did not change failure mode, and the fracture pattern was mainly favorable. More dentin walls need to be retained in clinic. When no less than two walls remained, a fiber post is not always necessary.


Mortality from lymphohematopoietic malignancies among workers in formaldehyde industries: the National Cancer Institute Cohort.
Formaldehyde exposure is associated with leukemia in some epidemiological studies. In the National Cancer Institute's formaldehyde cohort, previously followed through December 31, 1979, and updated through December 31, 1994, formaldehyde exposure was associated with an increased risk for leukemia, particularly myeloid leukemia, that increased with peak and average intensity of exposure. We extended follow-up through December 31, 2004 (median follow-up = 42 years), for 25 619 workers employed at one of 10 formaldehyde-using or formaldehyde-producing plants before 1966. We used Poisson regression to calculate relative risk (RR) estimates and 95% confidence intervals (CIs) to examine associations between quantitative formaldehyde exposure estimates (peak exposure, average intensity and cumulative exposure) and death from lymphohematopoietic malignancies. All statistical tests were two-sided and considered to be significant at P = .05. When follow-up ended in 2004, there were statistically significant increased risks for the highest vs lowest peak formaldehyde exposure category (> or =4 parts per million [ppm] vs >0 to <2.0 ppm) and all lymphohematopoietic malignancies (RR = 1.37; 95% CI = 1.03 to 1.81, P trend = .02) and Hodgkin lymphoma (RR = 3.96; 95% CI = 1.31 to 12.02, P trend = .01). Statistically nonsignificant associations were observed for multiple myeloma (RR = 2.04; 95% CI = 1.01 to 4.12, P trend > .50), all leukemia (RR = 1.42; 95% CI = 0.92 to 2.18, P trend = .12), and myeloid leukemia (RR = 1.78; 95% CI = 0.87 to 3.64, P trend = .13). There was little evidence of association for any lymphohematopoietic malignancy with average intensity or cumulative exposure at the end of follow-up in 2004. However, disease associations varied over time. For peak exposure, the highest formaldehyde-related risks for myeloid leukemia occurred before 1980, but trend tests attained statistical significance in 1990 only. After the mid-1990s, the formaldehyde-related risk of myeloid leukemia declined. Evaluation of risks over time suggests a possible link between formaldehyde exposure and lymphohematopoietic malignancies, particularly myeloid leukemia but also perhaps Hodgkin lymphoma and multiple myeloma. Observed patterns could be due to chance but are also consistent with a causal association within the relatively short induction-incubation periods characteristic of leukemogenesis. Further epidemiological study and exploration of potential molecular mechanisms are warranted.


Depression and its Relationship to Work Status and Income Among HIV Clients in Uganda.
Despite high levels of depression among persons living with HIV (PLWHIV), little research has investigated the relationship of depression to work status and income in PLWHIV in sub-Saharan Africa, which was the focus of this analysis. Baseline data from a prospective longitudinal cohort of 798 HIV patients starting antiretroviral therapy in Kampala, Uganda were examined. In separate multivariate analyses, we examined whether depressive severity and symptom type [as measured by the Patient Health Questionnaire (PHQ-9)] and major depression [diagnosed with the Mini International Neuropsychiatric Interview (MINI)] were associated with work status and income, controlling for demographics, physical health functioning, work self-efficacy, social support and internalized HIV stigma. 14% of the sample had Major Depression and 66% were currently working. Each measure of depression (PHQ-9 total score, somatic and cognitive subscales; Major Depression diagnosis) was associated with not working and lower average weekly income in bivariate analysis. However, none of the depression measures remained associated with work and income in multivariate analyses that controlled for other variables associated with these economic outcomes. These findings suggest that while depression is related to work and income, its influence may only be indirect through its relationship to other factors such as work self-efficacy and physical health functioning.


Clinical evaluation of a novel bipolar radiofrequency ablation system for renal masses.
What's known on the subject? and What does the study add? With the advancement of minimally invasive surgery, the management of small renal masses (SRM) has dramatically changed. Ablative technology such as radiofrequency ablation (RFA) and cryoablation have emerged as viable alternative modalities to extirpative surgery. RFA is one of the most studied and applied energy-based, needle-ablative treatment modalities, with encouraging mid- and long-term oncological outcomes. Monopolar devices have several shortcomings. The electrodes are susceptible to the cooling effect of nearby blood vessels that act as a 'heat sink', limiting the extent of tissue ablation and forming lesions with asymmetric borders and 'skip lesions'. Therefore, it is difficult to monitor and accurately predict the size of ablated lesions. A novel bipolar radiofrequency ablation (BRFA) device has been recently developed to address concerns with monopolar systems (Trod Medical, Paris, France). The BRFA system addresses the limitations of monopolar RFA, in terms of lesion size, targeting, consistency and concerns about cell death in the ablated area. We evaluated the BRFA device in 10 patients undergoing laparoscopic partial or radical nephrectomy. The present study demonstrates the safety and efficacy of a novel BRFA device. A BRFA device can produce a defined reproducible lesion with a precise transition zone to normal tissue. The area of ablated tissue exhibited completely devitalized cells and precise transition zone. With these characteristics, the potential advantages of this new technology during RFA ablation of SRM include less collateral damage and more complete ablation without skip lesions. This has the potential to lower rates of local recurrence and reduce incidence of skin burns. Further follow-up studies are necessary to determine its oncological efficacy. To evaluate a novel bipolar radiofrequency ablation (BRFA) system for the destruction of kidney tumours in patients. Bipolar radiofrequency ablation (BRFA) was used to ablate renal masses in 10 patients undergoing laparoscopic radical or partial nephrectomy. The probe was placed percutaneously and laparoscopically guided into the tumour after routine laparoscopic exposure. The electrical current was continuously adjusted by the generator to overcome disruption from increasing impedance created from desiccated tissue. The specimens were then excised in routine fashion and analysed by a single pathologist. Lesion size and shape, and size of the transition zone to viable tissue were measured via nicotinamide adenine dinucleotide (NADH) staining. Ablation was successful in all 10 tumours. Mean time to set up and place the probe was between 2 and 4 min. Duration of ablation was 200 s. None of the ablated tissue showed signs of viable cells by histological examination and NADH staining. The mean size of the ablation zone was 6.26 cm(3), with regular borders and a tapered cylindrical shape similar to the shape of the outer coil. The width of the transition zone, or area spanning complete tissue ablation to the first viable cells, ranged from 10 to 60 µm. There were no complications noted due to the ablation. A BRFA device can produce a defined reproducible lesion with a precise transition zone to normal tissue. The area of ablated tissue exhibited completely devitalized cells and precise transition zone.


[Autism and hyperlexia].
Hyperlexia is described in children who present alterations or retardation in development and is frequently characterised by their teaching themselves to read at an early age. This ability to decode words does not correlate with their intellectual level and can course with different degrees of mental retardation. It is always accompanied by difficulty in establishing social relationships. Autism is a disorder that is prototypical of the autistic spectrum disorders (ASD), which are essentially characterised by qualitative alterations in social interaction, in communication and language development, and in the presence of a limited repertory of interests, accompanied by stereotyped or peculiar responses. The presence of hyperlexia in autistic children is more frequent than in other development disorders. This phenomenon is linked with the increased skills involving visual memory, visual discrimination and motivation/interest towards visually represented material that is to be found in people with autism. However, not all autistic individuals present hyperlexia and not all children with hyperlexia present autism, although evidence shows that hyperlexia is a phenomenon that is observed with greater frequency in autism and in ASD. Hyperlexia, understood as meaning a little island of ability in children with autism and ASD, poses a number of questions and represents an important challenge in neuropsychological research in this population.


Identification of novel sweet protein for nutritional applications.
The prevalence of obesity and diabetes has increased exponentially in recent years around the globe, especially in India. Sweet proteins have the potential to substitute the sugars, by acting as natural, good and low calorie sweeteners. They also do not trigger a demand for insulin in diabetic patients unlike sucrose. In humans, the sweet taste perception is mainly due to taste-specific G protein-coupled heterodimeric receptors T1R2-T1R3. These receptors recognize diverse natural and synthetic sweeteners such as monelin, brazzein, thaumatin, curculin, mabinlin, miraculin and pentadin. Structural modeling of new sweetener proteins will be a great leap in further advancement of knowledge and their utility as sweeteners. We have explored the fingerprints of sweetness by studying the aminoacid composition and structure properties of the above proteins. The structural analysis of monellin revealed that the individual A or B chains of monellin are not contributing to its sweetness. However, the native conformation and ionic interaction between AspB7 of monellin with active site of T1R2-T1R3 receptor, along with hydrogen bonding stability of IleB6 and IleB8 are responsible for the sweet taste. Based on structural similarity search, we found a new hypothetical protein from Shewanella loihica, which has the presence of Asp(32) with adjacent isoleucine residues. Further, we examined the lead protein by two-step docking for the study of interaction of functionally conserved residues with receptors. The identified protein showed similar ionic and hydrophobic interactions with monelin. This gives a promising opportunity to explore this protein for potential health application in the low calorie sweetener industry viz., soft drinks, snacks, food, chocolate industries etc.


Predominance of CD8+ T lymphocytes in the inflammatory lesions of mice with acute Trypanosoma cruzi infection.
Using glycol methacrylate in conjunction with avidin-biotin-peroxidase complex techniques, we studied the contribution of T cell subsets to tissue inflammation during acute Trypanosoma cruzi infection. Two mouse/parasite model systems whose parasitology and pathology behaved differently were used. In C57Bl/6J mice infected with the T. cruzi Brazil strain, the levels of parasitism in blood and tissue (myocardial and skeletal muscle) reached a maximum at week 6 and decreased rapidly thereafter. Inflammatory responses in tissue corresponded with the parasitism, but decreased in intensity more gradually than that of parasitism. The T lymphocytes (Thy 1.2+) were found to be the major lymphocyte population in inflammatory cardiac and skeletal muscles (64.6-81.2%) at both three and six weeks postinfection. Among T cells, CD8+ cells (47.0-58.9%) significantly outnumbered CD4+ cells (9.3-18.6%). The number of B cells (0-1.0%) and macrophages was low. Experiments using C3H/HeSnJ mice infected with the Sylvio X10/4 clone of T. cruzi at 30 days postinfection resulted in similar findings except for a higher CD8+:CD4+ ratio. The primary finding of this study is that Thy 1.2+CD8+CD4- T lymphocytes are the major cell population in both heart and skeletal muscle in acute murine T. cruzi infection. The predominance of CD8+ T cells coincident with the decrease in the tissue parasite burden suggests a role for CD8+ T cells in the control of T. cruzi at the level of the infected cell.


Community tobacco control leaders' perceptions of harm reduction.
To investigate community tobacco control leaders' attitudes toward harm reduction approaches to tobacco use, in order to assess benefits and risks associated with these strategies. Cross sectional design involving qualitative outcomes from nine structured focus groups. 47 community tobacco control leaders in Minnesota working in the areas of public policy, clinical treatment of nicotine dependence and youth development participated. Participants discussed definitions of harm reduction; benefits and risks of harm reduction methods; and how funds for tobacco control research and programmes should be allocated. Results indicated inconsistency about the definition of harm reduction: most groups included a broad range of strategies that extended beyond those typically referenced in the scientific literature. Many participants stated that harm reduction might be beneficial, particularly for smokers who could not or would not quit. However, most also expressed concern about a number of risks, including delivering a mixed message about tobacco, inadvertently benefiting the tobacco industry, and causing unanticipated negative health effects. Participants were inclined to suggest public policy measures (for example, smoking bans, increased taxes) as means for reducing harm. Results indicate that even among tobacco control leaders there is a need for common terminology to describe harm reduction approaches and that public policy approaches to harm reduction are considered more dependable than strategies that involve pharmaceutical treatment or rely on the tobacco industry, such as product modification.


Systemic administration of human adipose-derived stem cells reverts nociceptive hypersensitivity in an experimental model of neuropathy.
Over the last decade, it has been proved that mesenchymal stem cells (MSCs) elicit anti-inflammatory effects. MSCs from adipose tissue (hASCs) differentiate into cells of the mesodermal lineage and transdifferentiate into ectodermal-origin cells. Although there are various etiologies to chronic pain, one common feature is that painful states are associated with increased inflammation. We believe in hASCs as a therapeutic tool also in pathologies involving neuroinflammation and neuronal tissue damage. We have investigated the effect of hASCs injected in a model of neuropathic pain [(mouse sciatic nerve chronic constriction injury (CCI)]. hASCs from 5 donors were characterized, and no major differences were depicted. hASCs were cryopreserved and grown on demand. About 1×10(6), 3×10(6), and 6×10(6) hASCs were intravenously injected into normal immunocompetent mice. No mouse died, and no macroscopic toxicity or behavioral changes were observed, confirming the safety of hASCs. hASCs, intravenously (i.v.) injected into C57BL/6 mice when the neuropathic pain was already established, induced a significant reduction in mechanical allodynia and a complete reversion of thermal hyperalgesia in a dose-response fashion, already 1 day after administration. Moreover, the hASCs effect can be boosted by repeated administrations, allowing a prolonged therapeutic effect. Treatment decreased the level of the CCI-induced proinflammatory cytokine interleukin (IL)-1β and activated the anti-inflammatory cytokine IL-10 in the lesioned nerve. hASCs treatment also restored normal inducible nitric oxide synthase expression in the spinal cord of CCI animals. Our data suggest that hASCs are worthy of further studies as an anti-inflammatory therapy in the treatment of neuropathic pain or chronic inflammatory diseases.


A metagenomic survey of microbes in honey bee colony collapse disorder.
In colony collapse disorder (CCD), honey bee colonies inexplicably lose their workers. CCD has resulted in a loss of 50 to 90% of colonies in beekeeping operations across the United States. The observation that irradiated combs from affected colonies can be repopulated with naive bees suggests that infection may contribute to CCD. We used an unbiased metagenomic approach to survey microflora in CCD hives, normal hives, and imported royal jelly. Candidate pathogens were screened for significance of association with CCD by the examination of samples collected from several sites over a period of 3 years. One organism, Israeli acute paralysis virus of bees, was strongly correlated with CCD.


Effect of Stabilizers on Encapsulation Efficiency and Release Behavior of Exenatide-Loaded PLGA Microsphere Prepared by the W/O/W Solvent Evaporation Method.
The aim of this study was to investigate the effects of various stabilizers on the encapsulation efficiency and release of exenatide-loaded PLGA (poly(lactic-co-glycolic acid)) microspheres prepared by the water-in-oil-in-water (W/O/W) solvent evaporation (SE) method. It was shown that the stabilizers affected exenatide stability in aqueous solutions, at water/dichloromethane interfaces, on PLGA surfaces, or during freeze-thawing and freeze-drying procedures. Sucrose predominantly reduces instability generated during freeze-thawing and freeze-drying. Phenylalanine prevents the destabilization at the water-dichloromethane (DCM) interface through decreased adsorption. Poloxamer 188 enhances stability in aqueous solutions and prevents adsorption to PLGA. Proline and lysine decrease adsorption on PLGA surfaces. Fourier transform infra-red spectroscopy (FT-IR) was used to find the molecular interaction of additives with exenatide or PLGA. Additives used in stability assessments were then added stepwise into the inner or outer water phase of the W/O/W double emulsion, and exenatide-loaded microspheres were prepared using the solvent evaporation method. The effect of each stabilizer on the encapsulation efficiency and release behavior of microspheres correlated well with the stability assessment results, except for the negative effect of poloxamer 188. Particle size analysis using laser diffractometry, scanning electron microscopy (SEM), water vapor sorption analysis, differential scanning calorimetry (DSC), and circular dichroism (CD) spectroscopy were also employed to characterize the prepared exenatide-loaded PLGA microsphere. This study demonstrated that an adequate formulation can be obtained by the study about the effect of stabilizers on peptide stability at the preformulation step. In addition, it can help to overcome various problems that can cause the destabilization of a peptide during the microsphere-manufacturing process and sustained drug release.


Temporally-regulated retinoic acid depletion produces specific neural crest, ocular and nervous system defects.
Both retinoid receptor null mutants and classic nutritional deficiency studies have demonstrated that retinoids are essential for the normal development of diverse embryonic structures (e.g. eye, heart, nervous system, urogenital tract). Detailed analysis of retinoid-modulated events is hampered by several limitations of these models, including that deficiency or null mutation is present throughout gestation, making it difficult to isolate primary effects, and preventing analysis beyond embryolethality. We developed a mammalian model in which retinoid-dependent events are documented during distinct targeted windows of embryogenesis. This was accomplished through the production of vitamin A-depleted (VAD) female rats maintained on sufficient oral retinoic acid (RA) for growth and fertility. After mating to normal males, these RA-sufficient/VAD females were given oral RA doses which allowed for gestation in an RA-sufficient state; embryogenesis proceeded normally until retinoids were withdrawn dietarily to produce a sudden, acute retinoid deficiency during a selected gestational window. In this trial, final RA doses were administered on E11.5, vehicle at E12.5, and embryos analyzed on E13.5; during this 48 hour window, the last RA dose was metabolized and embryos progressed in a retinoid-deficient state. RA-sufficient embryos were normal. Retinoid-depleted embryos exhibited specific malformations of the face, neural crest, eyes, heart, and nervous system. Some defects were phenocopies of those seen in null mutant mice for RXR alpha(-/-), RXR alpha(-/-)/RAR alpha(-/-), and RAR alpha(-/-)/RAR gamma(-/-), confirming that RA transactivation of its nuclear receptors is essential for normal embryogenesis. Other defects were unique to this deficiency model, showing that complete ligand 'knock-out' is required to see those retinoid-dependent events previously concealed by receptor functional redundancy, and reinforcing that retinoid receptors have separate yet overlapping contributions in the embryo. This model allows for precise targeting of retinoid form and deficiency to specific developmental windows, and will facilitate studies of distinct temporal events.


Phosphate monoester hydrolysis by trinuclear alkaline phosphatase; DFT study of transition States and reaction mechanism.
Alkaline phosphatase (AP) is a trinuclear metalloenzyme that catalyzes the hydrolysis of a broad range of phosphate monoesters to form inorganic phosphate and alcohol (or phenol). In this paper, by using density functional theory with a model based on a crystal structure, the AP-catalyzed hydrolysis of phosphate monoesters is investigated by calculating two substrates, that is, methyl and p-nitrophenyl phosphates, which represent alkyl and aryl phosphates, respectively. The calculations confirm that the AP reaction employs a "ping-pong" mechanism involving two chemical displacement steps, that is, the displacement of the substrate leaving group by a Ser102 alkoxide and the hydrolysis of the phosphoseryl intermediate by a Zn2-bound hydroxide. Both displacement steps proceed via a concerted associative pathway no matter which substrate is used. Other mechanistic aspects are also studied. Comparison of our calculations with linear free energy relationships experiments shows good agreement.


Biomechanical comparison of augmentation techniques for insufficiency fractures.
Biomechanical laboratory research. To compare 4 techniques for vertebral body insufficiency fracture repair by analyzing restoration of height, strength, and stiffness, and quantifying cement leakage. Current techniques for insufficiency fracture repair, restore prefracture strength, and all potentially have the complication of cement extrusion. To our knowledge, there is no information directly comparing different augmentation techniques. There were 4 osteoporotic/osteopenic fresh cadaveric spines' (T6-L5) vertebral bodies divided into the 4 groups and by levels (thoracic, thoracolumbar, lumbar). They were compressed to 25% of their initial height to simulate insufficiency fractures. Fixation techniques were vertebroplasty, osteoplasty (Interpore Cross Int., Irvine, CA), the Cavity Creation System (Synthes, Paoli, PA), and kyphoplasty (Kyphon, Sunnyvale, CA). Fractures were fixed using 3 cc of bone cement and recompressed. No significant difference was found in strength, height restoration, or stiffness among techniques (P < 0.05). Stiffness was significantly different in the lumbar levels between initial and post-fixation measurements (P < 0.05). Cement extrusion of 1 mL was seen in 1 vertebroplasty specimen. The 4 techniques equally restored strength and stiffness. No difference was seen in height restoration. Cement extrusion was significant in only 1 vertebral body, which may be attributable to the cement quantity.


Somatic and vicarious pain are represented by dissociable multivariate brain patterns.
Understanding how humans represent others' pain is critical for understanding pro-social behavior. 'Shared experience' theories propose common brain representations for somatic and vicarious pain, but other evidence suggests that specialized circuits are required to experience others' suffering. Combining functional neuroimaging with multivariate pattern analyses, we identified dissociable patterns that predicted somatic (high versus low: 100%) and vicarious (high versus low: 100%) pain intensity in out-of-sample individuals. Critically, each pattern was at chance in predicting the other experience, demonstrating separate modifiability of both patterns. Somatotopy (upper versus lower limb: 93% accuracy for both conditions) was also distinct, located in somatosensory versus mentalizing-related circuits for somatic and vicarious pain, respectively. Two additional studies demonstrated the generalizability of the somatic pain pattern (which was originally developed on thermal pain) to mechanical and electrical pain, and also demonstrated the replicability of the somatic/vicarious dissociation. These findings suggest possible mechanisms underlying limitations in feeling others' pain, and present new, more specific, brain targets for studying pain empathy.


Characterization and reactivity of monoclonal antibodies to the Miller strain of transmissible gastroenteritis virus of swine.
Hybridomas secreting monoclonal (MAB) to transmissible gastroenteritis virus (TGEV) were produced by fusion of SP2/0 myeloma cells and splenic lymphocytes of BALB/c mice immunized with the virulent cell-passaged Miller strain of TGEV. The MAB secreted by these hybridomas were partially characterized; 4 of them (MA4, MA5, MH11, MB2) had high-neutralization titer for TGEV. The remaining 7 (MC6, MD9, ME5, MG5, MF2, ME9, MG7) did not neutralize TGEV at 1:25 dilution. All 4 neutralizing and 2 of the nonneutralizing MAB reacted with the E2 protein of TGEV in a radioimmunoprecipitation assay. The remaining 5 MAB reacted with the E1 protein of TGEV. Reactivity of the MAB was tested in an indirect immunofluorescent assay with 3 cell culture-adapted strains of TGEV (Miller, Purdue, and Illinois) and 13 wild-type isolates of TGEV. Neutralizing MAB reacted with all 13 wild-type isolates and the 3 cell culture-adapted strains of TGEV. In contrast, nonneutralizing MAB that reacted with the Miller strain of TGEV varied in their reactivity with the wild-type TGEV isolates. Reactivity of neutralizing MAB was also tested, using plaque-reduction neutralization assays with Miller, Purdue, and Illinois strains and 5 wild-type isolates. All 4 neutralizing MAB neutralized the 8 virus isolates, but the neutralization titer was higher with the homologous virus than with the heterologous virus isolates. However, neutralization titers of the 4 neutralizing MAB were 4 to 16 times higher for the homologous Miller strain of TGEV than for the heterologous Illinois and Purdue strains, and were 4 to 1,000 times higher than for the wild-type isolates.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of wettability on two-phase quasi-static displacement: Validation of two pore scale modeling approaches.
Understanding of pore-scale physics for multiphase flow in porous media is essential for accurate description of various flow phenomena. In particular, capillarity and wettability strongly influence capillary pressure-saturation and relative permeability relationships. Wettability is quantified by the contact angle of the fluid-fluid interface at the pore walls. In this work we focus on the non-trivial interface equilibria in presence of non-neutral wetting and complex geometries. We quantify the accuracy of a volume-of-fluid (VOF) formulation, implemented in a popular open-source computational fluid dynamics code, compared with a new formulation of a level set (LS) method, specifically developed for quasi-static capillarity-dominated displacement. The methods are tested in rhomboidal packings of spheres for a range of contact angles and for different rhomboidal configurations and the accuracy is evaluated against the semi-analytical solutions obtained by Mason and Morrow (1994). While the VOF method is implemented in a general purpose code that solves the full Navier-Stokes (NS) dynamics in a finite volume formulation, with additional terms to model surface tension, the LS method is optimized for the quasi-static case and, therefore, less computationally expensive. To overcome the shortcomings of the finite volume NS-VOF system for low capillary number flows, and its computational cost, we introduce an overdamped dynamics and a local time stepping to speed up the convergence to the steady state, for every given imposed pressure gradient (and therefore saturation condition). Despite these modifications, the methods fundamentally differ in the way they capture the interface, as well as in the number of equations solved and in the way the mean curvature (or equivalently capillary pressure) is computed. This study is intended to provide a rigorous validation study and gives important indications on the errors committed by these methods in solving more complex geometry and dynamics, where usually many sources of errors are interplaying.


Characteristics of anterior cruciate ligament injuries in Australian football.
Anterior cruciate ligament (ACL) injuries are the most costly injuries in football at both professional and amateur levels (Orchard J, Seward H, McGivern J, Hood S. Intrinsic and extrinsic risk factors for anterior cruciate ligament injury in Australian footballers. Am J Sports Med 2001;29:196-200.). In this study video analysis of 34 ACL injuries in Australian football was performed to investigate the causes of these injuries. Factors that may have contributed to the cause of the injury were analysed, rated and reported. The factors analysed were: type of manoeuvre, direction the knee 'gave way', running speed, knee angle, cutting angle and if the player was accelerating or decelerating. The majority of the injuries analysed occurred in non-contact situations (56%). Of these 37% occurred during sidestepping manoeuvres, 32% in landing, 16% land and step, 10% stopping/slowing and 5% crossover cut manoeuvres. Ninety-two percent of the non-contact injuries occurred at extended knee angles of 30 degrees or less, which is also commonly known to place stress on the ACL and reduce the protective role of hamstrings. Over half (54%) of non-contact injuries occurred whilst decelerating. It would be expected that greater speed and angle cut too would increase the frequency of ACL injury. The results could not confirm this with most injuries occurring at running speeds of slow jogging to running and equal number of injuries occurred at cutting to angles of the ranges 15-45 degrees and 45-75 degrees. These results give greater understanding into potential causes or contributors of ACL injury and information to assist in the development of knee injury prevention programs.


Occurrence and characteristics of extended-spectrum β-lactamase (ESBL) producing Enterobacteriaceae in food producing animals, minced meat and raw milk.
The impact of food animals as a possible reservoir for extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae, and the dissemination of such strains into the food production chain need to be assessed. In this study 334 fecal samples from pigs, cattle, chicken and sheep were investigated at slaughter. Additionally, 100 raw milk samples, representing bulk tank milk of 100 different dairy farms, 104 minced meat (pork and beef) samples and 67 E. coli isolates from cattle E. coli mastitis were analyzed. As many as 15.3% of the porcine, 13.7% of the bovine, 8.6% of the sheep and 63.4% of the chicken fecal samples yielded ESBL producers after an enrichment step. In contrast, none of the minced meat, none of the bulk tank milk samples and only one of the mastitis milk samples contained ESBL producing strains. Of the total of 91 isolates, 89 were E. coli, one was Citrobacter youngae and one was Enterobacter cloacae. PCR analysis revealed that 78 isolates (85.7%) produced CTX-M group 1 ESBLs while six isolates (6.6%) produced CTX-M group 9 enzymes. Five detected ESBLs (5.5%) belonged to the SHV group and 2 isolates (2.2%) contained a TEM-type enzyme. A total of 27 CTX-M producers were additionally PCR-positive for TEM-beta-lactamase. The ESBL-encoding genes of 53 isolates were sequenced of which 34 produced CTX-M-1, 6 produced CTX-M-14, 5 produced CTX-M-15 and also 5 produced SHV-12. Two isolates produced TEM-52 and one isolate expressed a novel CTX-M group 1 ESBL, CTX-M-117. One isolate--aside from a CTX-M ESBL-- contained an additional novel TEM-type broad-spectrum beta-lactamase, TEM-186. The relatively high rates of ESBL producers in food animals and the high genetic diversity among these isolates are worrisome and indicate an established reservoir in farm animals.


Effect of earthworm loads on organic matter and nutrient removal efficiencies in synthetic domestic wastewater, and on bacterial community structure and diversity in vermifiltration.
In this paper, we studied the effect of earthworm loads on the removal rates of chemical oxygen demand (COD), ammonia nitrogen (NH3-N), total nitrogen, and total phosphorus from synthetic domestic sewage and on the bacterial community structure and diversity of substrates in earthworm packing beds. The different vermifiltrations (VFs), including the control, are successful in removing both organic matter (OM) and nutrients. The removal rate of NH3-N at 12.5 g of earthworm/L of soil VF is higher compared with that at 0 and 4.5 earthworm load VFs. The highest Shannon index, in the earthworm packing bed, occurred at 16.5 earthworm load VF. Furthermore, the COD removal rate is significantly correlated with the Shannon index, which reveals that OM removal for synthetic domestic sewage treatment at VF might be more dependent on bacterial diversity at the earthworm packing bed. The band distributions and diversities of the bacterial community for samples from different earthworm loads in VFs suggest that the bacterial community structure was only affected within the earthworm packing bed when the earthworm load reached a certain level. The present study adds to the current understanding of OM and nutrient degradation processes in VF domestic wastewater treatment.


Efficacy of Bacillus thuringiensis israelensis, Bacillus sphaericus and temephos for managing Anopheles larvae in Eritrea.
We evaluated the larvicidal activity of the granular formulation of Bacillus thuringiensis israelensis (Bti) serotype H-14 (Vectobac G, 200 ITU/mg) and Bacillus sphaericus (Bsph) serotype H5a5b (Vectolex CG, 670 Bs ITU/mg) against Anopheles arabiensis and other mosquitoes in breeding habitats in 3 sites, Gash-Barka, Anseba, and Debub zones, in Eritrea. The primary objective was to determine the optimal application rate and duration of effect for Bti and Bsph in representative larval habitats as compared with the organophosphate temephos. The biolarvicides were tested at 100% (high) and 50% (low) of the maximum recommended application rate. Temephos was applied at a rate of 100 ml/ha. At least 4 replicate experiments with Vectobac G (5.6 and 11.2 kg/ha), Vectolex CG (11.2 and 22.4 kg/ha) were conducted in each study site. All 3 larvicides caused significant mortality of the main malaria vector species, An. arabiensis, and other mosquito species (Anopheles cinereus, Anopheles pretoriensis, Culex quinquefasciatus). The larvicidal activity for Bti and Bsph was variable depending upon breeding habitat, mosquito species, and general ecology of the area. Both biopesticides had a similar duration of activity (2-3 wk) and were generally as effective as temephos for these time periods. In some cases, the high and low application rates for Bti and Bsph produced equivalent control over 2-3 wk. The 2 Bacillus biopesticides were less effective in habitats with high algal content and in fast flowing streams primarily because of the inability to penetrate algal mats and dilution effect, respectively. The results show that application of the 2 biolarvicides bimonthly to streambed pools, rain pools, and similar habitats would maintain control of the anopheline mosquito population.


Genome evolution in major Escherichia coli O157:H7 lineages.
Genetic analysis of Escherichia coli O157:H7 strains has shown divergence into two distinct lineages, lineages I and II, that appear to have distinct ecological characteristics, with lineage I strains more commonly associated with human disease. In this study, microarray-based comparative genomic hybridization (CGH) was used to identify genomic differences among 31 E. coli O157:H7 strains that belong to various phage types (PTs) and different lineage-specific polymorphism assay (LSPA) types. A total of 4,084 out of 6,057 ORFs were detected in all E. coli O157:H7 strains and 1,751 were variably present or absent. Based on this data, E. coli O157:H7 strains were divided into three distinct clusters, which consisted of 15 lineage I (LSPA type 111111), four lineage I/II (designated in this study) (LSPA type 211111) and 12 lineage II strains (LSPA 222222, 222211, 222212, and 222221), respectively. Eleven different genomic regions that were dominant in lineage I strains (present in > or =80% of lineage I and absent from > or = 92% of lineage II strains) spanned segments containing as few as two and up to 25 ORFs each. These regions were identified within E. coli Sakai S-loops # 14, 16, 69, 72, 78, 83, 85, 153 and 286, Sakai phage 10 (S-loops # 91, 92 and 93) and a genomic backbone region. All four lineage I/II strains were of PT 2 and possessed eight of these 11 lineage I-dominant loci. Several differences in virulence-associated loci were noted between lineage I and lineage II strains, including divergence within S-loop 69, which encodes Shiga toxin 2, and absence of the non-LEE encoded effector genes nleF and nleH1-2 and the perC homologue gene pchD in lineage II strains. CGH data suggest the existence of two dominant lineages as well as LSPA type and PT-related subgroups within E. coli O157:H7. The genomic composition of these subgroups supports the phylogeny that has been inferred from other methods and further suggests that genomic divergence from an ancestral form and lateral gene transfer have contributed to their evolution. The genomic features identified in this study may contribute to apparent differences in the epidemiology and ecology of strains of different E. coli O157:H7 lineages.


Gender-specific, multi-level determinants of outcomes of antihypertensive treatment: a sub-analysis of the Belgian PREVIEW study.
Gender-specific determinants of blood pressure (BP) values and control have not been the focus of clinical hypertension research. The purpose of this analysis was to identify gender-specific and multi-level (physician and patient) determinants of BP values and predictors of uncontrolled BP. We completed a subgroup analysis comparing men and women who participated in the Belgian PREVIEW study of second-line treatment effectiveness of valsartan, applying two-level hierarchical modelling of 90-day BP values and guideline-defined BP control. In total, 1665 women and 1525 men were treated by 504 general practitioners. Fewer women than men reached systolic BP (SBP) (P=0.015) and combined BP targets at 90 days (P=0.007). More than 26% of the variance in 90-day SBP (intra-class correlation coefficient (ICC)=0.270) and diastolic BP (DBP) (ICC=0.262) was attributable to physician-level factors for men; the physician-level ICCs for SBP and DBP were 0.259 and 0.268, respectively, for women. Determinants of 90-day BP values and predictors of uncontrolled BP varied considerably by gender. Many of the multi-level determinants of BP by gender are amenable to intervention, and the remainder can serve as warning signs to clinicians that patients may remain vulnerable to poor outcomes associated with sub-optimal BP control.


Relationship between incidence of gastric dilatation-volvulus and biometeorologic events in a population of military working dogs.
To calculate the monthly incidence of gastric dilatation-volvulus (GDV) in a population of military working dogs during a 5-year period and determine whether there was an association with synoptic climatologic indices. Medical records of all military working dogs housed at Lackland Air Force Base,Tex, from Jan 1, 1993 to Dec 31, 1997. Confirmed cases of GDV were identified from evaluation of medical records and used to calculate incidence of GDV. Factor analysis of local climatologic data was used to classify each day into 1 of 8 meteorologically homogeneous types of days for this location. Occurrence of GDV was compared with frequency of occurrence of synoptic climatologic days. 48 cases of GDV were identified from January 1993 through December 1997 Mean monthly incidence was 2.5 cases/1,000 dogs at risk (range, 0 to 18.5 cases/1,000 dogs; median, 2.5 cases/1,000 dogs). A seasonal increase in incidence of GDV was detected, because half of the episodes were during November, December, and January. An association with a specific synoptic climatologic day was not detected. Seasonal fluctuations in incidence of GDV may be associated with external factors that precipitate physiologic changes resulting in GDV. Although a specific cause-effect relationship was not documented, clinicians must be alert for the potential of seasonal variation in incidence of GDV and accordingly heighten their index of suspicion for the condition, particularly in populations of dogs that are predisposed to development of GDV.


Quantifying the disparity in outcome between urban and rural patients with acute appendicitis in South Africa.
Acute appendicitis in South Africa is associated with higher morbidity than in the developed world. Objective. To compare outcomes of urban and rural patients in KwaZulu-Natal and to determine whether there are disparities in outcome. We conducted a prospective study from September 2010 to September 2012 at Edendale Hospital in Pietermaritzburg, South Africa. All patients who presented with acute appendicitis were included. The operative and clinical course of urban and rural patients was compared. Results. A total of 500 patients were included, with 200 patients in the rural group and 300 in the urban group. Those from the rural group had a significantly longer duration of symptoms prior to presentation. All septic parameters were significantly worse in the rural group. Significantly more patients from the rural group required a laparotomy (77% v. 51% urban; p<0.001). Inflamed, non-perforated appendicitis was more commonly seen in the urban group (52.3% v. 21% rural; p<0.001), while perforated appendicitis was much more common in the rural group (79% v. 47.7% urban; p<0.001). Perforation associated with generalised, four-quadrant intra-abdominal contamination was significantly higher in the rural group than the urban group (60.5% v. 21%, respectively; p<0.05). Significantly more patients from the rural group required an open abdomen (46% v. 12% urban; p<0.001) and ≥1 re-laparotomies to control severe intra-abdominal sepsis (60.5% v. 23.3% urban; p<0.001). We have identified rural origin as an independent indicator of poor outcome. Possible reasons may include difficulty in accessing the health system or delay in transfer to a regional hospital. These need to be investigated further.


Ecotourism and primate habituation: Behavioral variation in two groups of white-faced capuchins (Cebus capucinus) from Costa Rica.
The increase of ecotourism operations within Costa Rica during the last 20 yrs has brought more and more humans into close, direct contact with several wildlife species. One of these species is the white-faced capuchin (Cebus capucinos), highly gregarious, and with exposure over time, willing to come into close vicinity of humans and their developments. Such contact has its advantages and disadvantages for the ecotourism industry. We observed white-faced monkeys in order to assess the impact of human presence and development on monkey behavior, with a focus on aggressive, affiliative, and foraging behaviors in Curú Wildlife Refuge (CWR), located in Puntarenas, Costa Rica, and to ascertain the degree of over-habituation of capuchin popula- tions at CWR. Though there exists no discrete behavioral parameters that measure over-habituation, it can be defined as an extreme state of habituation in which non-human primates not only lose fear of humans, but also actively include humans in social interactions or treat them as a food resource. We used instantaneous focal animal and group scan sampling during 8 wks in March and April 2012. Two groups (approximately 20-30 individuals each) of capuchins were observed; the first near the tourist development at the Southwestern area of CWR, representing a habituated population that regularly foraged, rested, and groomed in the presence of humans. The second, was observed in the Northeastern area of CWR, did not visit the center of human activity and exhibited fear of humans. The habituated group exhibited significantly fewer instances of threatened behavior in response to human presence (p < 0.0001) than the non-habituated group, and spent significantly more time eating and foraging (p < 0.0001). While the habituated monkeys at CWR may not be over-habituated, they could become that way as development, especially ecotourism, increases. Over-habituation is a problem that affects capuchins in certain ecotourism sites in Costa Rica. It is critical that the consequences of habituation be studied more carefully, primarily in areas where ecotourism operations draw visitors to wildlife habitats.


A case of neuroendocrine oncogenic osteomalacia associated with a PHEX and fibroblast growth factor-23 expressing sinusidal malignant schwannoma.
Oncogenic osteomalacia is a rare paraneoplastic syndrome that is characterized biochemically by hypophosphatemia and low plasma 1,25-dihydroxyvitamin D3, and clinically by osteomalacia, pseudofractures, bone pain, fatigue, and muscle weakness. We present a patient with a malignant schwannoma as the underlying cause of this disorder. A permanent cell line (HMS-97) derived from this tumor showed evidence of neuroendocrine differentiation by immunohistochemistry and of neurosecretory activity by electron microscopy. The cell line did express PHEX (phosphate-regulating gene with homologies to endopeptidases located on the X-chromosome) and FGF-23 (fibroblast growth factor-23) transcripts on northern hybridization; however, none of the known mutations from the related mendelian disorders of X-linked hypophosphatemic rickets or autosomal-dominant hypophosphatemic rickets could be detected. Tumor cell (HMS-97)-derived conditioned medium did not inhibit phosphate transport in a standard opossum kidney cell assay and in animal experiments. The medium also showed no PTH1- or PTH2-receptor-stimulating bioactivity. HMS-97 cells might be useful for further studies that aim to determine the genetic mechanism that leads to the observed PHEX and FGF-23 expression, both of which might have a direct role in the pathogenesis of oncogenic osteomalacia. In addition, these cells might be a useful tool for the investigation of neuroendocrine Schwann cell function and autoimmune peripheral nerve disease.


Prediction of overall survival for patients with metastatic castration-resistant prostate cancer: development of a prognostic model through a crowdsourced challenge with open clinical trial data.
Improvements to prognostic models in metastatic castration-resistant prostate cancer have the potential to augment clinical trial design and guide treatment strategies. In partnership with Project Data Sphere, a not-for-profit initiative allowing data from cancer clinical trials to be shared broadly with researchers, we designed an open-data, crowdsourced, DREAM (Dialogue for Reverse Engineering Assessments and Methods) challenge to not only identify a better prognostic model for prediction of survival in patients with metastatic castration-resistant prostate cancer but also engage a community of international data scientists to study this disease. Data from the comparator arms of four phase 3 clinical trials in first-line metastatic castration-resistant prostate cancer were obtained from Project Data Sphere, comprising 476 patients treated with docetaxel and prednisone from the ASCENT2 trial, 526 patients treated with docetaxel, prednisone, and placebo in the MAINSAIL trial, 598 patients treated with docetaxel, prednisone or prednisolone, and placebo in the VENICE trial, and 470 patients treated with docetaxel and placebo in the ENTHUSE 33 trial. Datasets consisting of more than 150 clinical variables were curated centrally, including demographics, laboratory values, medical history, lesion sites, and previous treatments. Data from ASCENT2, MAINSAIL, and VENICE were released publicly to be used as training data to predict the outcome of interest-namely, overall survival. Clinical data were also released for ENTHUSE 33, but data for outcome variables (overall survival and event status) were hidden from the challenge participants so that ENTHUSE 33 could be used for independent validation. Methods were evaluated using the integrated time-dependent area under the curve (iAUC). The reference model, based on eight clinical variables and a penalised Cox proportional-hazards model, was used to compare method performance. Further validation was done using data from a fifth trial-ENTHUSE M1-in which 266 patients with metastatic castration-resistant prostate cancer were treated with placebo alone. 50 independent methods were developed to predict overall survival and were evaluated through the DREAM challenge. The top performer was based on an ensemble of penalised Cox regression models (ePCR), which uniquely identified predictive interaction effects with immune biomarkers and markers of hepatic and renal function. Overall, ePCR outperformed all other methods (iAUC 0·791; Bayes factor >5) and surpassed the reference model (iAUC 0·743; Bayes factor >20). Both the ePCR model and reference models stratified patients in the ENTHUSE 33 trial into high-risk and low-risk groups with significantly different overall survival (ePCR: hazard ratio 3·32, 95% CI 2·39-4·62, p<0·0001; reference model: 2·56, 1·85-3·53, p<0·0001). The new model was validated further on the ENTHUSE M1 cohort with similarly high performance (iAUC 0·768). Meta-analysis across all methods confirmed previously identified predictive clinical variables and revealed aspartate aminotransferase as an important, albeit previously under-reported, prognostic biomarker. Novel prognostic factors were delineated, and the assessment of 50 methods developed by independent international teams establishes a benchmark for development of methods in the future. The results of this effort show that data-sharing, when combined with a crowdsourced challenge, is a robust and powerful framework to develop new prognostic models in advanced prostate cancer. Sanofi US Services, Project Data Sphere.


Overexpression of microRNA‑486 affects the proliferation and chemosensitivity of mesothelioma cell lines by targeting PIM1.
Dysregulated levels of microRNAs (miRNAs or miRs), involved in oncogenic pathways, have been proposed to contribute to the aggressiveness of malignant pleural mesothelioma (MPM). Previous studies have highlighted the downregulation of miRNA miR‑486‑5p in patients with mesothelioma and the introduction of miRNA mimics to restore their reduced or absent functionality in cancer cells is considered an important therapeutic strategy. The aim of the present study was to evaluate the mechanisms through which miRNAs may influence the functions, proliferation and sensitivity to cisplatin of MPM cells. In the present study, a miR‑486‑5p mimic was transfected into the H2052 and H28 MPM cell lines, and cell viability, proliferation, apoptosis and mitochondrial membrane potential were monitored. miR‑486‑5p overexpression led to a clear impairment of cell proliferation, targeting CDK4 and attenuating cell cycle progression. In addition, transfection with miR‑486‑5p mimic negatively regulated the release of inflammatory factors and the expression of Provirus integration site for Moloney murine leukaemia virus 1 (PIM1). The sensitivity of the cells to cisplatin was enhanced by enhancing the apoptotic effects of the drug and impairing mitochondrial function. On the whole, the present study demonstrates that miR‑486‑5p may play an important role in MPM treatment by targeting multiple pathways involved in tumour development and progression. These activities may be mostly related to the downregulation of PIM1, a crucial regulator of cell survival and proliferation. Furthermore, these results provide support for the combined use of miR‑486‑5p with chemotherapy as a therapeutic strategy for MPM.


Intracellular colocalization of variant surface glycoprotein and transferrin-gold in Trypanosoma brucei.
Endocytosis and intracellular transport has been studied in the bloodstream forms of Trypanosoma brucei by light and electron microscopy, using colloidal gold coupled to bovine transferrin (transferrin-gold). The endocytosed transferrin-gold, visualized by silver intensification for light microscopy, was present in vesicular structures between the cell nucleus and flagellar pocket of the organism. At the ultrastructural level, transferrin-gold was present after a 10-min incubation in the flagellar pocket, coated vesicles, cisternal networks, and lysosomelike structures. Endocytosis and intracellular processing of T. brucei variable surface glycoprotein (VSG) was studied using two preparations of affinity-purified rabbit IgG directed against different parts of the VSG. One preparation of IgG was directed against the cross-reacting determinant (CRD): a complex glycolipid side chain covalently linked to the COOH-terminus of the VSG molecule. The other was directed against determinants on the rest of the VSG molecule. When the two IgG preparations were used on thawed, thin cryosections of trypanosomes that had been incubated in transferrin-gold before fixation, the organelles involved with transferrin-gold endocytosis labeled with both antibodies, as well as many vesicular, tubular, and vacuolar structures that did not contain endocytosed transferrin-gold. Both antibodies also labeled the cell surface. In double-labeling experiments both antibodies were closely associated except that IgG directed against the VSG molecule labeled all the cisternae of the Golgi apparatus, whereas anti-CRD IgG was shown to label only half of the Golgi apparatus. Evidence for sorting of VSG molecules from endocytosed transferrin-gold was found. Double-labeling experiments also showed some tubular profiles which labeled on one side with anti-CRD IgG and on the other side with anti-VSG IgG, suggesting a possible segregation of parts of the VSG molecule.


Consciousness as a concrete physical phenomenon.
The typical empirical approach to studying consciousness holds that we can only observe the neural correlates of experiences, not the experiences themselves. In this paper we argue, in contrast, that experiences are concrete physical phenomena that can causally interact with other phenomena, including observers. Hence, experiences can be observed and scientifically modelled. We propose that the epistemic gap between an experience and a scientific model of its neural mechanisms stems from the fact that the model is merely a theoretical construct based on observations, and distinct from the concrete phenomenon it models, namely the experience itself. In this sense, there is a gap between any natural phenomenon and its scientific model. On this approach, a neuroscientific theory of the constitutive mechanisms of an experience is literally a model of the subjective experience itself. We argue that this metatheoretical framework provides a solid basis for the empirical study of consciousness.


Globin gene activation during haemopoiesis is driven by protein complexes nucleated by GATA-1 and GATA-2.
How does an emerging transcriptional programme regulate individual genes as stem cells undergo lineage commitment, differentiation and maturation? To answer this, we have analysed the dynamic protein/DNA interactions across 130 kb of chromatin containing the mouse alpha-globin cluster in cells representing all stages of differentiation from stem cells to mature erythroblasts. The alpha-gene cluster appears to be inert in pluripotent cells, but priming of expression begins in multipotent haemopoietic progenitors via GATA-2. In committed erythroid progenitors, GATA-2 is replaced by GATA-1 and binding is extended to additional sites including the alpha-globin promoters. Both GATA-1 and GATA-2 nucleate the binding of various protein complexes including SCL/LMO2/E2A/Ldb-1 and NF-E2. Changes in protein/DNA binding are accompanied by sequential alterations in long-range histone acetylation and methylation. The recruitment of polymerase II, which ultimately leads to a rapid increase in alpha-globin transcription, occurs late in maturation. These studies provide detailed evidence for the more general hypothesis that commitment and differentiation are primarily driven by the sequential appearance of key transcriptional factors, which bind chromatin at specific, high-affinity sites.


Optic Nerve Sheath as a Novel Mechanical Load on the Globe in Ocular Duction.
The optic nerve (ON) sheath's role in limiting duction has been previously unappreciated. This study employed magnetic resonance imaging (MRI) to demonstrate this constraint on adduction. High-resolution, surface coil axial MRI was obtained in 11 normal adults, 14 subjects with esotropia (ET) having normal axial length (AL) < 25.8 mm, 13 myopic subjects with ET and mean AL 29.3 ± 3.3 (SD) mm, and 7 subjects with exotropia (XT). Gaze angles and ON lengths were measured for scans employing eccentric lateral fixation in which an ON became completely straightened. In all groups, ON straightening occurred only in the adducting, not abducting, eye. Adduction at ON straightening was 26.0 ± 8.8° in normal subjects, not significantly different from XT at 22.2 ± 11.8°. However, there was significant increase in comparable adduction in ET to 36.3 ± 9.3°, and in myopic ET to 33.6 ± 10.7° (P < 0.04). Optic nerve length at straightening was 27.6 ± 2.7 mm in normals, not significantly different from 28.2 ± 2.8 mm in ET and 27.8 ± 2.7 mm in XT. In myopic ET, ON length at straightening was significantly reduced to 24.0 ± 2.9 mm (P < 0.002) and was associated with globe retraction in adduction, suggesting ON tethering. Large adduction may exhaust length redundancy in the normally sinuous ON and sheath, so that additional adduction must stretch the sheath and retract or deform the globe. These mechanical effects are most significant in ET with axial myopia, but may also exert traction on the posterior sclera absent strabismus or myopia. Tethering by the ON sheath in adduction is an important, novel mechanical load on the globe.


FMRP targets distinct mRNA sequence elements to regulate protein expression.
Fragile X syndrome (FXS) is a multi-organ disease that leads to mental retardation, macro-orchidism in males and premature ovarian insufficiency in female carriers. FXS is also a prominent monogenic disease associated with autism spectrum disorders (ASDs). FXS is typically caused by the loss of fragile X mental retardation 1 (FMR1) expression, which codes for the RNA-binding protein FMRP. Here we report the discovery of distinct RNA-recognition elements that correspond to the two independent RNA-binding domains of FMRP, in addition to the binding sites within the messenger RNA targets for wild-type and I304N mutant FMRP isoforms and the FMRP paralogues FXR1P and FXR2P (also known as FXR1 and FXR2). RNA-recognition-element frequency, ratio and distribution determine target mRNA association with FMRP. Among highly enriched targets, we identify many genes involved in ASD and show that FMRP affects their protein levels in human cell culture, mouse ovaries and human brain. Notably, we discovered that these targets are also dysregulated in Fmr1(-/-) mouse ovaries showing signs of premature follicular overdevelopment. These results indicate that FMRP targets share signalling pathways across different cellular contexts. As the importance of signalling pathways in both FXS and ASD is becoming increasingly apparent, our results provide a ranked list of genes as basis for the pursuit of new therapeutic targets for these neurological disorders.


The influence of patient and doctor gender on diagnosing coronary heart disease.
Using novel methods, this paper explores sources of uncertainty and gender bias in primary care doctors' diagnostic decision-making about coronary heart disease (CHD). Claims about gendered consultation styles and quality of care are re-examined, along with the adequacy of CHD models for women. Randomly selected doctors in the UK and the US (n=112, 56 per country, stratified by gender) were shown standardised videotaped vignettes of actors portraying patients with CHD. Patients' age, gender, ethnicity and social class were varied systematically. During interviews, doctors gave free-recall accounts of their decision-making, which were analysed to determine patient and doctor gender effects. We found differences in male and female doctors' responses to different types of patient information. Female doctors recall more patient cues overall, particularly about history presentation, and particularly amongst women. Male doctors appear less affected by patient gender but both male and especially female doctors take more account of male patients' age, and consider more age-related disease possibilities for men than women. Findings highlight the need for better integration of knowledge about female presentations within accepted CHD risk models, and do not support the contention that women receive better-quality care from female doctors.


The Future of Biosimilars: Maximizing Benefits Across Immune-Mediated Inflammatory Diseases.
Biologics have transformed the treatment of immune-mediated inflammatory diseases such as rheumatoid arthritis (RA) and inflammatory bowel disease (IBD). Biosimilars-biologic medicines with no clinically meaningful differences in safety or efficacy from licensed originators-can stimulate market competition and have the potential to expand patient access to biologics within the parameters of treatment recommendations. However, maximizing the benefits of biosimilars requires cooperation between multiple stakeholders. Regulators and developers should collaborate to ensure biosimilars reach patients rapidly without compromising stringent quality, safety, or efficacy standards. Pharmacoeconomic evaluations and payer policies should be updated following biosimilar market entry, minimizing the risk of imposing nonmedical barriers to biologic treatment. In RA, disparities between treatment guidelines and national reimbursement criteria could be addressed to ensure more uniform patient access to biologics and enable rheumatologists to effectively implement treat-to-target strategies. In IBD, the cost-effectiveness of biologic treatment earlier in the disease course is likely to improve when biosimilars are incorporated into pharmacoeconomic analyses. Patient understanding of biosimilars is crucial for treatment success and avoiding nocebo effects. Full understanding of biosimilars by physicians and carefully considered communication strategies can help support patients initiating or switching to biosimilars. Developers must operate efficiently to be sustainable, without undermining product quality, the reliability of the supply chain, or pharmacovigilance. Developers should also facilitate information sharing to meet the needs of other stakeholders. Such collaboration will help to ensure a sustainable future for both the biosimilar market and healthcare systems, supporting the availability of effective treatments for patients.


Participants' use of enacted scenes in research interviews: A method for reflexive analysis in health and social care.
In our study of a workforce intervention within a health and social care context we found that participants who took part in longitudinal research interviews were commonly enacting scenes from their work during one-to-one interviews. Scenes were defined as portions of the interviews in which participants directly quoted the speech of at least two actors. Our analysis in this paper focuses on these enacted scenes, and compares the content of them before and after the intervention. We found that, whilst the tensions between consistency and change, and change management, were common topics for scene enactment in both pre and post-intervention data, following the intervention participants were much more likely to present themselves as active agents in that change. Post-intervention enacted scenes also showed participants' reports of taking a service user perspective, and a focus on their interactions with service users that had been absent from pre-intervention data. In addition, descriptions of positive feeling and emotions were present in the post-intervention enacted scenes. We suggest that this analysis confirms the importance of enacted scenes as an analytic resource, and that this importance goes beyond their utility in identifying the impact of this specific intervention. Given the congruence between the themes prominent in enacted scenes, and those which emerged from a more extensive qualitative analysis of these data, we argue that enacted scenes may also be of wider methodological importance. The possibility of using scene enactment as an approach to the validation of inductive analysis in health and social care settings could provide a useful methodological resource in settings where longitudinal ethnographic observation of frontline care staff is impossible or impractical.


Mesenchymal stem cells induce resistance to chemotherapy through the release of platinum-induced fatty acids.
The development of resistance to chemotherapy is a major obstacle for lasting effective treatment of cancer. Here, we demonstrate that endogenous mesenchymal stem cells (MSCs) become activated during treatment with platinum analogs and secrete factors that protect tumor cells against a range of chemotherapeutics. Through a metabolomics approach, we identified two distinct platinum-induced polyunsaturated fatty acids (PIFAs), 12-oxo-5,8,10-heptadecatrienoic acid (KHT) and hexadeca-4,7,10,13-tetraenoic acid (16:4(n-3)), that in minute quantities induce resistance to a broad spectrum of chemotherapeutic agents. Interestingly, blocking central enzymes involved in the production of these PIFAs (cyclooxygenase-1 and thromboxane synthase) prevents MSC-induced resistance. Our findings show that MSCs are potent mediators of resistance to chemotherapy and reveal targets to enhance chemotherapy efficacy in patients.


Clinical patterns, natural history, and progression of ulcerative colitis. A long-term follow-up of 1116 patients.
We studied the natural history of ulcerative colitis (UC) by following 1116 patients in whom UC had been diagnosed or confirmed at The Cleveland Clinic Foundation between 1960 and 1983. Data before 1973 were obtained retrospectively. Criteria for inclusion in the study were: a diagnosis of UC confirmed by clinical, radiographic, endoscopic, and histologic examination; disease location that could be defined as one of three categories (proctosigmoiditis, pancolitis, or left-sided colitis); and a follow-up of at least five years (mean = 12.7 years). Mean age at diagnosis was 32 years. Of the 1116 patients, 46.2% (516) had proctosigmoiditis; 36.7% (410) had pancolitis (colitis of the entire large intestine); and 17.0% (190) had left-sided colitis (from the dentate line to the splenic flexure). Early complications (within two years of diagnosis) included colonic hemorrhage (16.7%) and toxic colitis (12.7%). Complications were highest among patients with pancolitis. Surgery was required for 37.6% of the patients. Primary indications for surgery included chronic or intractable disability (40.2%), fulminating medical failure (16.9%), and colonic dilatation (18.4%). At the most recent follow-up, the disease had extended (progressed to a more serious category) in 53.8% of the patients, although 67.2% were asymptomatic and only 37.0% were on medications. Both the final disease destination and the initial diagnosis impacted cumulative colectomy-ileostomy rates. Factors associated with extension were toxic colitis (P < 0.0001); extent of disease at diagnosis (P < 0.0001); joint symptoms (P = 0.0008); younger age at diagnosis (P = 0.06); and severe bleeding (P = 0.07).


Associations Between Objective Sleep and Ambulatory Blood Pressure in a Community Sample.
Epidemiologic data increasingly support sleep as a determinant of cardiovascular disease risk. Fewer studies have investigated the mechanisms underlying this relationship using objective sleep assessment approaches. Therefore, the aim of this study was to examine associations between daily blood pressure (BP) and both objectively assessed sleep duration and efficiency. A diverse community sample of 300 men and women aged 21 to 70 years, enrolled in the North Texas Heart Study, participated in the study. Actigraphy-assessed sleep was monitored for two consecutive nights with ambulatory BP sampled randomly within 45-minute blocks on the first and second day as well as the second night. Overall, sleep duration results paralleled those of sleep efficiency. Individuals with lower sleep efficiency had higher daytime systolic (B = -0.35, SE = 0.11, p = .0018, R = 0.26) but not diastolic BP (B = -0.043, SE = 0.068, p = .52, R = 0.17) and higher nighttime BP (systolic: B = -0.37, SE = 0.10, p < .001, R = .15; diastolic: B = -0.20, SE = 0.059, p < .001, R = .14). Moreover, lower sleep efficiency on one night was associated with higher systolic (B = -0.51, SE = 0.11, p < .001, R = 0.23) and diastolic BP (B = -0.17, SE = 0.065, p = .012, R = .16) the following day. When 'asleep' BP was taken into account instead of nighttime BP, the associations between sleep and BP disappeared. When both sleep duration and efficiency were assessed together, sleep efficiency was associated with daytime systolic BP, whereas sleep duration was associated with nighttime BP. Lower sleep duration and efficiency are associated with higher daytime systolic BP and higher nighttime BP when assessed separately. When assessed together, sleep duration and efficiency diverge in their associations with BP at different times of day. These results warrant further investigation of these possible pathways to disease.


Semi-automatic segmentation of multiple mouse embryos in MR images.
The motivation behind this paper is to aid the automatic phenotyping of mouse embryos, wherein multiple embryos embedded within a single tube were scanned using Magnetic Resonance Imaging (MRI). Our algorithm, a modified version of the simplex deformable model of Delingette, addresses various issues with deformable models including initialization and inability to adapt to boundary concavities. In addition, it proposes a novel technique for automatic collision detection of multiple objects which are being segmented simultaneously, hence avoiding major leaks into adjacent neighbouring structures. We address the initialization problem by introducing balloon forces which expand the initial spherical models close to the true boundaries of the embryos. This results in models which are less sensitive to initial minimum of two fold after each stage of deformation. To determine collision during segmentation, our unique collision detection algorithm finds the intersection between binary masks created from the deformed models after every few iterations of the deformation and modifies the segmentation parameters accordingly hence avoiding collision.We have segmented six tubes of three dimensional MR images of multiple mouse embryos using our modified deformable model algorithm. We have then validated the results of the our semi-automatic segmentation versus manual segmentation of the same embryos. Our Validation shows that except paws and tails we have been able to segment the mouse embryos with minor error. This paper describes our novel multiple object segmentation technique with collision detection using a modified deformable model algorithm. Further, it presents the results of segmenting magnetic resonance images of up to 32 mouse embryos stacked in one gel filled test tube and creating 32 individual masks.


Redistribution of synaptic efficacy supports stable pattern learning in neural networks.
Markram and Tsodyks, by showing that the elevated synaptic efficacy observed with single-pulse long-term potentiation (LTP) measurements disappears with higher-frequency test pulses, have critically challenged the conventional assumption that LTP reflects a general gain increase. This observed change in frequency dependence during synaptic potentiation is called redistribution of synaptic efficacy (RSE). RSE is here seen as the local realization of a global design principle in a neural network for pattern coding. The underlying computational model posits an adaptive threshold rather than a multiplicative weight as the elementary unit of long-term memory. A distributed instar learning law allows thresholds to increase only monotonically, but adaptation has a bidirectional effect on the model postsynaptic potential. At each synapse, threshold increases implement pattern selectivity via a frequency-dependent signal component, while a complementary frequency-independent component nonspecifically strengthens the path. This synaptic balance produces changes in frequency dependence that are robustly similar to those observed by Markram and Tsodyks. The network design therefore suggests a functional purpose for RSE, which, by helping to bound total memory change, supports a distributed coding scheme that is stable with fast as well as slow learning. Multiplicative weights have served as a cornerstone for models of physiological data and neural systems for decades. Although the model discussed here does not implement detailed physiology of synaptic transmission, its new learning laws operate in a network architecture that suggests how recently discovered synaptic computations such as RSE may help produce new network capabilities such as learning that is fast, stable, and distributed.


Partial Transformation of Imogolite by Decylphosphonic Acid Yields an Interface Active Composite Material.
The phosphonic acid moiety is commonly used as an anchoring group for the surface modification of imogolite. However, the impact of the reaction on its structure has never been clearly analyzed before. We study the reaction of imogolite and decylphosphonic acid by combining infrared spectroscopy, X-ray scattering, scanning electron microscopy, transmission electron microscopy, and solid-state nuclear magnetic resonance spectroscopy. Instead of a surface functionalization, we observe the formation of a lamellar phase interconnected with imogolite bundles. Although we find no evidence for grafted imogolite tubes, we observe the expected dispersion characteristics and stabilization of water in toluene emulsions described in the literature. Based on the surface chemistry of imogolite, we propose an explanation for the observed reactivity and link the structural features of the obtained composite material to its dispersibility in toluene and its observed properties at the toluene-water interface.


Global burden of diseases, injuries, and risk factors for young people's health during 1990-2013: a systematic analysis for the Global Burden of Disease Study 2013.
Young people's health has emerged as a neglected yet pressing issue in global development. Changing patterns of young people's health have the potential to undermine future population health as well as global economic development unless timely and effective strategies are put into place. We report the past, present, and anticipated burden of disease in young people aged 10-24 years from 1990 to 2013 using data on mortality, disability, injuries, and health risk factors. The Global Burden of Disease Study 2013 (GBD 2013) includes annual assessments for 188 countries from 1990 to 2013, covering 306 diseases and injuries, 1233 sequelae, and 79 risk factors. We used the comparative risk assessment approach to assess how much of the burden of disease reported in a given year can be attributed to past exposure to a risk. We estimated attributable burden by comparing observed health outcomes with those that would have been observed if an alternative or counterfactual level of exposure had occurred in the past. We applied the same method to previous years to allow comparisons from 1990 to 2013. We cross-tabulated the quantiles of disability-adjusted life-years (DALYs) by quintiles of DALYs annual increase from 1990 to 2013 to show rates of DALYs increase by burden. We used the GBD 2013 hierarchy of causes that organises 306 diseases and injuries into four levels of classification. Level one distinguishes three broad categories: first, communicable, maternal, neonatal, and nutritional disorders; second, non-communicable diseases; and third, injuries. Level two has 21 mutually exclusive and collectively exhaustive categories, level three has 163 categories, and level four has 254 categories. The leading causes of death in 2013 for young people aged 10-14 years were HIV/AIDS, road injuries, and drowning (25·2%), whereas transport injuries were the leading cause of death for ages 15-19 years (14·2%) and 20-24 years (15·6%). Maternal disorders were the highest cause of death for young women aged 20-24 years (17·1%) and the fourth highest for girls aged 15-19 years (11·5%) in 2013. Unsafe sex as a risk factor for DALYs increased from the 13th rank to the second for both sexes aged 15-19 years from 1990 to 2013. Alcohol misuse was the highest risk factor for DALYs (7·0% overall, 10·5% for males, and 2·7% for females) for young people aged 20-24 years, whereas drug use accounted for 2·7% (3·3% for males and 2·0% for females). The contribution of risk factors varied between and within countries. For example, for ages 20-24 years, drug use was highest in Qatar and accounted for 4·9% of DALYs, followed by 4·8% in the United Arab Emirates, whereas alcohol use was highest in Russia and accounted for 21·4%, followed by 21·0% in Belarus. Alcohol accounted for 9·0% (ranging from 4·2% in Hong Kong to 11·3% in Shandong) in China and 11·6% (ranging from 10·1% in Aguascalientes to 14·9% in Chihuahua) of DALYs in Mexico for young people aged 20-24 years. Alcohol and drug use in those aged 10-24 years had an annual rate of change of >1·0% from 1990 to 2013 and accounted for more than 3·1% of DALYs. Our findings call for increased efforts to improve health and reduce the burden of disease and risks for diseases in later life in young people. Moreover, because of the large variations between countries in risks and burden, a global approach to improve health during this important period of life will fail unless the particularities of each country are taken into account. Finally, our results call for a strategy to overcome the financial and technical barriers to adequately capture young people's health risk factors and their determinants in health information systems. Bill & Melinda Gates Foundation.


Polarization-dependent surface plasmon-driven catalytic reaction on a single nanowire monitored by SERS.
The polarizing effect of an excitation laser on a plasmon-driven catalytic reaction on a single nanowire (NW) was investigated experimentally and theoretically. The dimerization of 4-nitrobenzenethiol (4NBT) to p,p'-dimercaptoazobenzene (DMAB) due to localized surface plasmon resonance (LSPR) was realized and monitored via surface enhanced Raman scattering (SERS). The SERS signal degradation has been compensated by using different equivalent points on the NW. It was shown that the SERS signals of both the reactant and product were sensitive to the angles (θ) between the longitude of the NW and the polarization direction of the excitation laser. When the polarization is along the transverse direction of the NW, the SERS signals are drastically enhanced by the LSPR. The efficiency of the plasmon-driven catalytic reaction increased significantly. The mechanism of the polarization-dependent plasmon-driven catalytic reaction was revealed by our dark field experiment and numerical finite-difference time-domain simulation. It was demonstrated that the maximum intensity of the electric field near the surface of the NW would also be a function of the angle θ. The theoretical and experimental results were consistent with each other. This research may pave a way for controlling plasmon-driven catalytic reactions by changing the polarization of an excitation laser incident on single anisotropic nanostructures such as a single NW.


Biofilm Matrix Composition Affects the Susceptibility of Food Associated Staphylococci to Cleaning and Disinfection Agents.
Staphylococci are frequently isolated from food processing environments, and it has been speculated whether survival after cleaning and disinfection with benzalkonium chloride (BC)-containing disinfectants is due to biofilm formation, matrix composition, or BC efflux mechanisms. Out of 35 food associated staphylococci, eight produced biofilm in a microtiter plate assay and were identified as Staphylococcus capitis (2), S. cohnii, S. epidermidis, S. lentus (2), and S. saprophyticus (2). The eight biofilm producing strains were characterized using whole genome sequencing. Three of these strains contained the ica operon responsible for production of a polysaccharide matrix, and formed a biofilm which was detached upon exposure to the polysaccharide degrading enzyme Dispersin B, but not Proteinase K or trypsin. These strains were more tolerant to the lethal effect of BC both in suspension and biofilm than the remaining five biofilm producing strains. The five BC susceptible strains were characterized by lack of the ica operon, and their biofilms were detached by Proteinase K or trypsin, but not Dispersin B, indicating that proteins were major structural components of their biofilm matrix. Several novel cell wall anchored repeat domain proteins with domain structures similar to that of MSCRAMM adhesins were identified in the genomes of these strains, potentially representing novel mechanisms of ica-independent biofilm accumulation. Biofilms from all strains showed similar levels of detachment after exposure to alkaline chlorine, which is used for cleaning in the food industry. Strains with qac genes encoding BC efflux pumps could grow at higher concentrations of BC than strains without these genes, but no differences were observed at biocidal concentrations. In conclusion, the biofilm matrix of food associated staphylococci varies with respect to protein or polysaccharide nature, and this may affect the sensitivity toward a commonly used disinfectant.


Novel avulsion pattern of the left principal bronchus with involvement of the carina and caudal thoracic trachea in a cat.
A 2-year-old, 4.5 kg, neutered male domestic shorthair cat was presented to the emergency service with dyspnoea, anorexia and apathetic behaviour. Thoracic radiographs showed typical signs for a thoracic trauma and a tracheal lesion in the region of the carina, consistent with pseudoairway formation. Computed tomography (CT) was performed in the conscious cat to avoid aggravation of air leakage associated with ventilation. The additional CT findings were consistent with a novel pattern of a traumatic avulsion of the left principal bronchus expanding into the carina and caudal thoracic trachea. Despite the complex avulsion pattern, successful treatment was achieved surgically by performing an end-to-end anastomosis via a fifth right intercostal lateral thoracotomy. The cat was ventilated with a feeding tube and jet ventilation throughout. The cat showed excellent recovery 6 months after surgery.


Anthracyclines, small-molecule inhibitors of hypoxia-inducible factor-1 alpha activation.
Hypoxia-inducible factor-1 (HIF-1) is a central mediator of cellular responses to low oxygen and has recently become an important therapeutic target for solid tumor therapy. To identify small molecule inhibitors of the HIF-1 transcriptional activation, we have established a high through-put assay system using a stable transformant of mammalian cells that express the luciferase reporter gene construct containing a HIF-1 binding site. Using this system, we screened 5000 cultured broths of microorganisms, and we found that cinerubin (1-hydroxy aclacinomycin B) showed a significant inhibition of the reporter activity induced by hypoxic conditions. In addition, we demonstrated that aclarubicin also inhibited the HIF-1 transcriptional activity under hypoxic conditions, but neither doxorubicin nor daunorubicin inhibited it. Consistent with these results, cinerubin and aclarubicin inhibited the hypoxic induction of the vascular endothelial growth factor (VEGF) protein in HepG2 cells, but neither doxorubicin nor daunorubicin affected it. Thus, our results suggested that some anthracyclines are also acting as angiogenesis inhibitors.


Multiprotein bridging factor 1 (MBF1) is an evolutionarily conserved transcriptional coactivator that connects a regulatory factor and TATA element-binding protein.
Multiprotein bridging factor 1 (MBF1) is a transcriptional cofactor that bridges between the TATA box-binding protein (TBP) and the Drosophila melanogaster nuclear hormone receptor FTZ-F1 or its silkworm counterpart BmFTZ-F1. A cDNA clone encoding MBF1 was isolated from the silkworm Bombyx mori whose sequence predicts a basic protein consisting of 146 amino acids. Bacterially expressed recombinant MBF1 is functional in interactions with TBP and a positive cofactor MBF2. The recombinant MBF1 also makes a direct contact with FTZ-F1 through the C-terminal region of the FTZ-F1 DNA-binding domain and stimulates the FTZ-F1 binding to its recognition site. The central region of MBF1 (residues 35-113) is essential for the binding of FTZ-F1, MBF2, and TBP. When the recombinant MBF1 was added to a HeLa cell nuclear extract in the presence of MBF2 and FTZ622 bearing the FTZ-F1 DNA-binding domain, it supported selective transcriptional activation of the fushi tarazu gene as natural MBF1 did. Mutations disrupting the binding of FTZ622 to DNA or MBF1, or a MBF2 mutation disrupting the binding to MBF1, all abolished the selective activation of transcription. These results suggest that tethering of the positive cofactor MBF2 to a FTZ-F1-binding site through FTZ-F1 and MBF1 is essential for the binding site-dependent activation of transcription. A homology search in the databases revealed that the deduced amino acid sequence of MBF1 is conserved across species from yeast to human.