Effects of in vivo benzo(a)pyrene treatment on liver microsomal mixed-function oxidase activities of gilthead seabream (Sparus aurata).
Benzo(a)pyrene [B(a)P] treatment of gilthead seabream, 25 mg/kg, i.p. for 5 consecutive days, did not cause any significant changes in ethylmorphine N-demethylase and aniline 4-hydroxylase activities of liver microsomes. The same treatment did not alter the liver microsomal cytochrome b5 content, NADH-cytochrome b5 reductase and NADPH-cytochrome P450 reductase activities. However, benzo(a)pyrene treatment caused a 2-3-fold increase in 7-ethoxyresorufin O-deethylase (7-EROD) activity of gilthead seabream liver microsomes. Although, upon treatment, total cytochrome P450 content of liver microsomes increased about 1.7-fold in 1990 fall, no such increase was observed in spring 1991. However, a new cytochrome P450 with an apparent M(r) of 58,000 was observed on SDS-PAGE of liver microsomes obtained from benzo(a)pyrene treated gilthead seabream. Besides, in vitro addition of 0.2 x 10(-6) M benzo(a)pyrene to the incubation mixture inhibited 7-ethoxyresorufin O-deethylase activity by 93%. Gilthead seabream liver microsomal 4-ethoxyresorufin O-deethylase activity was characterized with respect to substrate concentration, amount of enzyme, type of buffer used, incubation period and temperature.


[Marine science in Revista de Biologia Tropical in its 50th anniversary].
The first paper published in the Revista de Biología Tropical (RBT) on anything related to marine science was in 1963. Since then the number of marine-related papers has increased to 637, which represents 27% of the total production of RBT (excluding the Supplements), and 33% since 1979. Most publications are Full Articles on Ecology (135 papers). The marine ecosystem of which there is more publications is the coral reefs (28); and fish is the most studied taxonomic group (165). Almost half of the Supplements are marine related (12). The RBT must continue its efforts to maintain itself as a leading marine science publication in Latin America.


Early diagnosis of perioperative myocardial infarction after coronary bypass grafting: a study using biomarkers and cardiac magnetic resonance imaging.
Myocardial injury related to coronary artery bypass grafting (CABG) is poorly characterized, and understanding the characteristic release of biomarkers associated with revascularization injury might provide novel therapeutic opportunities. This study characterized early changes in biomarkers after revascularization injury during on-pump CABG. This prospective study comprised 28 patients undergoing on-pump CABG and late gadolinium enhancement (LGE) cardiac magnetic resonance imaging (CMRI) who underwent measurements of cardiac troponin I (cTnI), creatine kinase-MB, and inflammatory markers (C-reactive protein, serum amyloid A, myeloperoxidase, interleukin 6, tumor necrosis factor-α, matrix metalloproteinase 9a, monocyte chemotactic protein-1, plasminogen activator inhibitor-1a) at baseline, at 1, 6, 12, and 24 hours, and at 1 week (inflammatory markers only) post-CABG. Biomarker results at 1 hour were studied for a relationship to new myocardial infarction as defined by CMRI-LGE, and the diagnostic utility of combining positive biomarkers was assessed. All patients had an uneventful recovery, but 9 showed a new myocardial infarction demonstrated by new areas of hyperenhancement on CMR. Peak cTnI at 24 hours (ρ = 0.66, p < 0.001) and CK-MB (ρ = 0.66, p < 0.001) correlated with the amount of new LGE. At 1 hour, 3 biomarkers--cTnI, interleukin 6, and tumor necrosis factor-α--were significantly elevated in patients with vs those without new LGE. Receiver operating curve analysis showed cTnI was the most accurate at detecting new LGE at 1 hour: a cutoff of cTnI exceeding 5 μg/L at 1 hour had 67% sensitivity and 79% specificity for detecting new LGE. Unexpected CABG-related myocardial injury occurs in a significant proportion of patients. A cTnI test at 1 hour after CABG could potentially differentiate patients with significant revascularization injury.


From risky to safer home care: health care assistants striving to overcome a lack of training, supervision, and support.
Patients receiving home care are becoming increasingly dependent upon competent caregivers' 24-h availability due to their substantial care needs, often with advanced care and home care technology included. In Sweden, care is often carried out by municipality-employed paraprofessionals such as health care assistants (HC assistants) with limited or no health care training, performing advanced care without formal training or support. The aim of this study was to investigate the work experience of the HC assistants and to explore how they manage when delivering 24-h home care to patients with substantial care needs. Grounded theory methodology involving multiple data sources comprising interviews with HC assistants (n=19) and field observations in patients' homes was used to collect data and constant comparative analysis was used for analysis. The initial analysis revealed a number of barriers, competence gap; trapped in the home setting; poor supervision and unconnected to the patient care system, describing the risks associated with the situations of HC assistants working in home care, thus affecting their working conditions as well as the patient care. The core process identified was the HC assistants' strivings to combine safe home care with good working conditions by using compensatory processes. The four identified compensatory processes were: day-by-day learning; balancing relations with the patient; self-managing; and navigating the patient care system. By actively employing the compensatory processes, the HC assistants could be said to adopt an inclusive approach, by compensating for their own barriers as well as those of their colleagues' and taking overall responsibility for their workplace. In conclusion, the importance of supporting HC assistants in relation to their needs for training, supervision,and support from health care professionals must be addressed when organising 24-h home care to patients with substantial care needs in the future.


Screening procedure from cattle feces and the prevalence of Escherichia coli O157:H7 in Taiwan dairy cattle.
A procedure has been established for screening Escherichia coli O157:H7 from bovine feces. It consists of four steps: enrichment, selective culture, phenotyping and genotyping. Modified trypticase soy broth (mTSB) containing 20 microg/mL of novobiocin was used for the enrichment step. The selective culture step was done using sorbitol MacConkey agar containing 0.05 microg/mL of cefixime and 2.5 microg/mL of potassium tellurite. The phenotyping steps include: species confirmation as E. coli, serotyping to O157 and H7, and the detection of verocytotoxin (VT) production. Genotyping was made up of confirmation of the presence of the O157 antigen (rfb), the H7 antigen (ficC), the adherence factor (eaeA), hemolysin (hlyA), and VT production (vt I and vt II) by polymerase chain reaction. mTSB gave better enrichment for E. coli O157 than gram-negative (GN) broth. The detection limit for this screening procedure was 0.85 +/- 0.5 cfu/g. Using the screening procedure described above, E. coli O157 was found in four out of 3062 (0.13%) fresh bovine fecal specimens, and in two out of 78 (2.56%) dairy herds in Taiwan. Three out of the four detected strains were vt II producing E. coli O157:H7, while the other was E. coli O157:H7 which lacked VT-producing ability.


An atlas of cortical arealization identifies dynamic molecular signatures.
The human brain is subdivided into distinct anatomical structures, including the neocortex, which in turn encompasses dozens of distinct specialized cortical areas. Early morphogenetic gradients are known to establish early brain regions and cortical areas, but how early patterns result in finer and more discrete spatial differences remains poorly understood1. Here we use single-cell RNA sequencing to profile ten major brain structures and six neocortical areas during peak neurogenesis and early gliogenesis. Within the neocortex, we find that early in the second trimester, a large number of genes are differentially expressed across distinct cortical areas in all cell types, including radial glia, the neural progenitors of the cortex. However, the abundance of areal transcriptomic signatures increases as radial glia differentiate into intermediate progenitor cells and ultimately give rise to excitatory neurons. Using an automated, multiplexed single-molecule fluorescent in situ hybridization approach, we find that laminar gene-expression patterns are highly dynamic across cortical regions. Together, our data suggest that early cortical areal patterning is defined by strong, mutually exclusive frontal and occipital gene-expression signatures, with resulting gradients giving rise to the specification of areas between these two poles throughout successive developmental timepoints.


To what extent do single symptoms from a depression rating scale predict risk of long-term sickness absence among employees who are free of clinical depression?
Depression rating scales have predicted long-term sickness absence (LTSA) in previous studies. With this study, we investigated to what extent single symptoms from a depression rating scale predicted LTSA among employees who were free of clinical depression. We studied 6,670 female employees in the Danish eldercare sector. Frequency of 12 depressive symptoms over the last 2 weeks was assessed with the Major Depression Inventory. A symptom was considered as elevated if it was present at least "slightly more than half of the time." Data were linked to a national register on LTSA (≥3 weeks). We calculated hazard ratios (HR) from Cox's proportional hazard models to analyze whether a symptom predicted time to onset of LTSA during a 1-year follow-up. Analyses were adjusted for age, family status, health behaviors, occupational group, and previous LTSA. Of the 12 symptoms, three predicted LTSA after adjustment for covariates: "felt low in spirits and sad" (HR = 1.41, 95 % CI = 1.05-1.89), "felt lacking in energy and strength" (HR = 1.33, 95 % CI = 1.08-1.64), and "had trouble sleeping at night" (HR = 1.38, 95 % CI = 1.09-1.74). Among female eldercare workers free of clinical depression, feelings of low spirits and sadness, feelings of lack of energy and strength, and sleep disturbances predict risk of LTSA. Interventions that decrease the prevalence of these symptoms might contribute to a reduction in LTSA in this population.


Aurolysis of alpha-C-deprotonated group 6 aminocarbene and thiocarbene complexes with Ph3PAu+.
Deprotonated Fischer-type aminocarbene complexes, (CO)5M=C(NR2)CH3 (M = Cr or W; R = Me or propyl), react with Ph3PAu+ by metal group substitution - (CO)5M for Ph3PAu+--and attachment of the extricated M(CO)5 to the deprotonated methyl group. (The products may also be seen as aminovinylgold compounds coordinated to M(CO)5 moieties.) DFT calculations at the B3LYP level of theory using model compounds indicate a clear preference of the gold unit for central C to terminal coordination in the ligand [NMe2CCH2]-, whereas the Cr(CO)5 has a 7 kcal mol(-1) preference for C(vinyl) coordination compared to N-coordination. In related thiocarbenes, the sulfur donor atom should be the preferred point of attachment for the metal carbonyl unit. The latter prediction is borne out in practice, and in the three products isolated, including Ph3PAu{C(=CH2)SPh}Cr(CO)5 in a mixed crystal with [Ph3PAuSPh]Cr(CO)5, precisely this coordination mode is present. The latter component of the mixed crystal has also been prepared independently of the vinyl one.


Epimerisation of chiral hydroxylactones by short-chain dehydrogenases/reductases accounts for sex pheromone evolution in Nasonia.
Males of all species of the parasitic wasp genus Nasonia use (4R,5S)-5-hydroxy-4-decanolide (RS) as component of their sex pheromone while only N. vitripennis (Nv), employs additionally (4R,5R)-5-hydroxy-4-decanolide (RR). Three genes coding for the NAD+-dependent short-chain dehydrogenases/reductases (SDRs) NV10127, NV10128, and NV10129 are linked to the ability of Nv to produce RR. Here we show by assaying recombinant enzymes that SDRs from both Nv and N. giraulti (Ng), the latter a species with only RS in the pheromone, epimerise RS into RR and vice versa with (4R)-5-oxo-4-decanolide as an intermediate. Nv-derived SDR orthologues generally had higher epimerisation rates, which were also influenced by NAD+ availability. Semiquantitative protein analyses of the pheromone glands by tandem mass spectrometry revealed that NV10127 as well as NV10128 and/or NV10129 were more abundant in Nv compared to Ng. We conclude that the interplay of differential expression patterns and SDR epimerisation rates on the ancestral pheromone component RS accounts for the evolution of a novel pheromone phenotype in Nv.


Inflammatory lymphangiogenesis in postpartum breast tissue remodeling.
Like many cancers, mammary carcinomas use lymphatic vessels to disseminate, and numerous clinical and experimental studies have documented a strong correlation between peritumoral lymphangiogenesis and tumor dissemination. At the same time, many other factors can affect the incidence, invasiveness, and mortality of breast cancer, including lactation history. Although lactation reduces overall cancer risk, patients diagnosed within 5 years of pregnancy have an increased incidence of metastatic disease. In this issue of the JCI, Lyons and colleagues demonstrate that postpartum breast tissue remodeling during involution coincides with inflammatory lymphangiogenesis. In mouse models, cyclooxygenase-2 (COX-2) inhibition during involution reduced the risk of cancer metastasis and correlated with decreased lymphangiogenesis. In addition to lymphangiogenesis, COX-2 inhibition reduces many of the immune-suppressive features of the tumor microenvironment, including development of myeloid-derived suppressor cells and regulatory T cells; therefore, these results support the notion that inhibiting COX-2 during lactation weaning may lessen the incidence of breast cancer metastasis.


Identification of atomic-level mechanisms for gas-phase X- + CH3Y SN2 reactions by combined experiments and simulations.
For the traditional model of gas-phase X(-) + CH3Y SN2 reactions, C3v ion-dipole pre- and postreaction complexes X(-)---CH3Y and XCH3---Y(-), separated by a central barrier, are formed. Statistical intramolecular dynamics are assumed for these complexes, so that their unimolecular rate constants are given by RRKM theory. Both previous simulations and experiments have shown that the dynamics of these complexes are not statistical and of interest is how these nonstatistical dynamics affect the SN2 rate constant. This work also found there was a transition from an indirect, nonstatistical, complex forming mechanism, to a direct mechanism, as either the vibrational and/or relative translational energy of the reactants was increased. The current Account reviews recent collaborative studies involving molecular beam ion-imaging experiments and direct (on-the-fly) dynamics simulations of the SN2 reactions for which Cl(-), F(-), and OH(-) react with CH3I. Also considered are reactions of the microsolvated anions OH(-)(H2O) and OH(-)(H2O)2 with CH3I. These studies have provided a detailed understanding of the atomistic mechanisms for these SN2 reactions. Overall, the atomistic dynamics for the Cl(-) + CH3I SN2 reaction follows those found in previous studies. The reaction is indirect, complex forming at low reactant collision energies, and then there is a transition to direct reaction between 0.2 and 0.4 eV. The direct reaction may occur by rebound mechanism, in which the ClCH3 product rebounds backward from the I(-) product or a stripping mechanism in which Cl(-) strips CH3 from the I atom and scatters in the forward direction. A similar indirect to direct mechanistic transition was observed in previous work for the Cl(-) + CH3Cl and Cl(-) + CH3Br SN2 reactions. At the high collision energy of 1.9 eV, a new indirect mechanism, called the roundabout, was discovered. For the F(-) + CH3I reaction, there is not a transition from indirect to direct reaction as Erel is increased. The indirect mechanism, with prereaction complex formation, is important at all the Erel investigated, contributing up ∼60% of the reaction. The remaining direct reaction occurs by the rebound and stripping mechanisms. Though the potential energy curve for the OH(-) + CH3I reaction is similar to that for F(-) + CH3I, the two reactions have different dynamics. They are akin, in that for both there is not a transition from an indirect to direct reaction. However, for F(-) + CH3I indirect reaction dominates at all Erel, but it is less important for OH(-) + CH3I and becomes negligible as Erel is increased. Stripping is a minor channel for F(-) + CH3I, but accounts for more than 60% of the OH(-) + CH3I reaction at high Erel. Adding one or two H2O molecules to OH(-) alters the reaction dynamics from that for unsolvated OH(-). Adding one H2O molecule enhances indirect reaction at low Erel, and changes the reaction mechanism from primarily stripping to rebound at high Erel. With two H2O molecules the dynamics is indirect and isotropic at all collision energies.


Cost-Effective Fabrication of Fractal Silicon Nanowire Arrays.
Silicon nanowires (Si NWs) emerged in several application fields as a strategic element to surpass the bulk limits with a flat compatible architecture. The approaches used for the Si NW realization have a crucial impact on their final performances and their final cost. This makes the research on a novel and flexible approach for Si NW fabrication a crucial point for Si NW-based devices. In this work, the novelty is the study of the flexibility of thin film metal-assisted chemical etching (MACE) for the fabrication of Si NWs with the possibility of realizing different doped Si NWs, and even a longitudinal heterojunction p-n inside the same single wire. This point has never been reported by using thin metal film MACE. In particular, we will show how this approach permits one to obtain a high density of vertically aligned Si NWs with the same doping of the substrate and without any particular constraint on doping type and level. Fractal arrays of Si NWs can be fabricated without any type of mask thanks to the self-assembly of gold at percolative conditions. This Si NW fractal array can be used as a substrate to realize controllable artificial fractals, integrating other interesting elements with a cost-effective microelectronics compatible approach.


Voyaging canoes and the settlement of polynesia.
Sailing trials with two reconstructed Polynesian double canoes indicate that these craft can make good a course to windward up to approximately 75 degrees off the wind on long ocean voyages. This windward performance would have enabled Polynesians to exert a degree of control over their movements that would have been denied them had they only been able to sail or drift before wind and current. Indeed, without this windward sailing capacity there probably never would have been a Polynesian people today, for in a sense they are a product of their maritime technology. Had there been no great voyaging canoes, the settlement of Polynesia might have had to await the relatively late entry into the Pacific of the European navigators. But the Pacific was the scene of early innovation in weatherly sailing canoes, and as the European navigators "discovered" island after island, they were surprised to find that neolithic seafarers had preceded them into this vast ocean realm.


Predictive Metagenomic Profiling, Urine Metabolomics, and Human Marker Gene Expression as an Integrated Approach to Study Alopecia Areata.
Involvement of the microbiome in many different scalp conditions has been investigated over the years. Studies on the role of the scalp microbiome in specific diseases, such as those involving hair growth alterations like non-cicatricial [androgenetic alopecia (AGA), alopecia areata (AA)] and cicatricial alopecia lichen planopilaris, are of major importance. In the present work, we highlighted the differences in microbial populations inhabiting the scalp of AA subjects and a healthy sample cohort by using an integrated approach relying on metagenomic targeted 16S sequencing analysis, urine metabolomics, and human marker gene expression. Significant differences in genera abundances (p < 0.05) were found in the hypodermis and especially the dermis layer. Based on 16S sequencing data, we explored the differences in predicted KEGG pathways and identified some significant differences in predicted pathways related to the AA pathologic condition such as flagellar, assembly, bacterial chemotaxis, mineral absorption, ABC transporters, cellular antigens, glycosaminoglycan degradation, lysosome, sphingolipid metabolism, cell division, protein digestion and absorption, and energy metabolism. All predicted pathways were significantly enhanced in AA samples compared to expression in healthy samples, with the exceptions of mineral absorption, and ABC transporters. We also determined the expression of TNF-α, FAS, KCNA3, NOD-2, and SOD-2 genes and explored the relationships between human gene expression levels and microbiome composition by Pearson's correlation analysis; here, significant correlations both positive (SOD vs. Staphylococcus, Candidatus Aquiluna) and negative (FAS and SOD2 vs. Anaerococcus, Neisseria, and Acinetobacter) were highlighted. Finally, we inspected volatile organic metabolite profiles in urinary samples and detected statistically significant differences (menthol, methanethiol, dihydrodehydro-beta-ionone, 2,5-dimethylfuran, 1,2,3,4, tetrahydro-1,5,7-trimethylnapthalene) when comparing AA and healthy subject groups. This multiple comparison approach highlighted potential traits associated with AA and their relationship with the microbiota inhabiting the scalp, opening up novel therapeutic interventions in such kind of hair growth disorders mainly by means of prebiotics, probiotics, and postbiotics.


Adsorption separation of carbon dioxide from flue gas by a molecularly imprinted adsorbent.
CO2 separation by molecularly imprinted adsorbent from coal-fired flue gas after desulfurization system has been studied. The adsorbent was synthesized by molecular imprinted technique, using ethanedioic acid, acrylamide, and ethylene glycol dimethacrylate as the template, functional monomer, and cross-linker, respectively. According to the conditions of coal-fired flue gas, the influencing factors, including adsorption temperature, desorption temperature, gas flow rate, and concentrations of CO2, H2O, O2, SO2, and NO, were studied by fixed bed breakthrough experiments. The experimental conditions were optimized to gain the best adsorption performance and reduce unnecessary energy consumption in future practical use. The optimized adsorption temperature, desorption temperature, concentrations of CO2, and gas flow rate are 60 °C, 80 °C, 13%, and 170 mL/min, respectively, which correspond to conditions of practical flue gases to the most extent. The CO2 adsorption performance was nearly unaffected by H2O, O2, and NO in the flue gas, and was promoted by SO2 within the emission limit stipulated in the Chinese emission standards of air pollutants for a thermal power plant. The maximum CO2 adsorption capacity, 0.57 mmol/g, was obtained under the optimized experimental conditions, and the SO2 concentration was 150 mg/m(3). The influence mechanisms of H2O, O2, SO2, and NO on CO2 adsorption capacity were investigated by infrared spectroscopic analysis.


Modular recurrent neural networks for Mandarin syllable recognition.
A new modular recurrent neural network (MRNN)- based speech-recognition method that can recognize the entire vocabulary of 1280 highly confusable Mandarin syllables is proposed in this paper. The basic idea is to first split the complicated task, in both feature and temporal domains, into several much simpler subtasks involving subsyllable and tone discrimination, and then to use two weighting RNN's to generate several dynamic weighting functions to integrate the subsolutions into a complete solution. The novelty of the proposed method lies mainly in the use of appropriate a priori linguistic knowledge of simple initial-final structures of Mandarin syllables in the architecture design of the MRNN. The resulting MRNN is therefore effective and efficient in discriminating among highly confusable Mandarin syllables. Thus both the time-alignment and scaling problems of the ANN-based approach for large-vocabulary speech-recognition can be addressed. Experimental results show that the proposed method and its extensions, the reverse-time MRNN (Rev-MRNN) and bidirection MRNN (Bi-MRNN), all outperform an advanced HMM method trained with the MCE/GPD algorithm in both recognition-rate and system complexity.


Effects of chitosan on human periodontal ligament fibroblasts in vitro and on bone formation in rat calvarial defects.
The purpose of this study was to evaluate the effect of chitosan on human periodontal ligament fibroblasts (hPDLF) in vitro and on bone formation in rat calvarial defects in vivo. Fibroblast populations were obtained from individuals with a healthy periodontium and cultured in alpha minimum essential medium (MEM) for the control group. For the experimental groups, cells were cultured in alpha-MEM containing chitosan at concentrations of 0.01, 0.1, 1, or 2 mg/ml. The 3-(4,5-dimethyl-thiazole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, reverse transcription-polymerase chain reaction (RT-PCR) and the assay of alkaline phosphatase (ALPase) activity were performed. Eight mm calvarial critical-sized defects were created in 30 male Sprague-Dawley rats. The animals were divided into three groups of 10 animals each. The defects were treated with either chitosan/absorbable collagen sponge (ACS) or ACS alone in the experimental groups or were left untreated (surgical controls). The animals were sacrificed at 2 or 8 weeks post-surgery and the treatment outcomes were evaluated using histological and histomorphometric parameters. The chitosan-induced proliferative responses of the hPDLF reached a plateau at a concentration of 0.1 mg/ml (P <0.05). When the hPDLF were stimulated with 0.1 mg/ml chitosan, both the mRNA expression of type I collagen and the ALP activity were significantly up-regulated (P <0.05). The surgical implantation of chitosan/ACS enhanced the new bone formation at 8 weeks post-surgery and the amount of new bone formation of the chitosan/ACS group was significantly greater than that of both the ACS alone group and the surgical control group (P <0.01). The new bone area and defect closure in the chitosan/ACS group were significantly greater than those in the ACS control and sham surgery control groups at 8 weeks (P <0.01). However, the chitosan/ ACS group exhibited significantly less bone density than both the ACS control and the sham surgery control group at 8 weeks (P <0.01). Chitosan (0.1 mg/ml) enhanced the type I collagen synthesis and facilitated the differentiation into osteogenic cells. Chitosan reconstituted with ACS has a significant potential to accelerate the regeneration of bone in rat calvarial critical size defects.


Reliability and validity of the Biodex system 3 pro isokinetic dynamometer velocity, torque and position measurements.
This study quantitatively assessed the mechanical reliability and validity of position, torque and velocity measurements of the Biodex System 3 isokinetic dynamometer. Trial-to-trial and day-to-day reliability were assessed during three trials on two separate days. To assess instrument validity, measurement of each variable using the Biodex System 3 dynamometer was compared to a criterion measure of position, torque and velocity. Position was assessed at 5 degrees increments across the available range of motion of the dynamometer. Torque measures were assessed isometrically by hanging six different calibrated weights from the lever arm. Velocity was assessed (30 degrees/s to 500 degrees/s) across a 70 degrees arc of motion by manually accelerating the weighted lever arm. With the exception of a systematic decrease in velocity at speeds of 300 degrees/s and higher, the Biodex System 3 performed with acceptable mechanical reliability and validity on all variables tested.


Simplified Disease Activity Index remission at month 6 is an independent predictor of functional and structural remissions at month 12 during abatacept treatment in patients with rheumatoid arthritis: A multi-center, prospective cohort study in Japan.
To evaluate association of clinical remission at month 6 with functional and structural remissions at month 12 during abatacept treatment in patients with rheumatoid arthritis (RA). This 12-month prospective, multicenter cohort study enrolled 168 patients with RA who started abatacept. Outcomes were assessed using composite measures, quality of life indices, and the van der Heijde-modified total Sharp score (mTSS). The logistic regression analysis was applied to identify factors associated with outcomes and their odds ratios (OR) with 95% confidence interval (95% CI). At month 6 and 12, 21.4% and 26.2% of the patients achieved Simplified Disease Activity Index (SDAI) remission (SDAI <3.3), and 40.6% and 41.7% achieved Health Assessment Questionnaire-Disability Index (HAQ-DI <0.5) remission. Among 129 patients whose mTSS progression was evaluated at month 12, 83 (64.3%) achieved structural remission (ΔmTSS ≤0.5 for 12 months). SDAI remission at month 6 was identified as a significant predictor of both functional (OR, 3.732; 95% CI, 1.328-10.489) and structural remissions (OR, 4.301; 95% CI, 1.298-14.243) at month 12 after adjusting for covariates. Aiming for SDAI remission at month 6 is an appropriate strategy to obtain good functional and structural outcomes at month 12.


High fat diets and mouse colon mucosal membranes: a centrifugation study.
Female Swiss mice were fed on a control diet (laboratory chow, 6% fat) or high fat diets (laboratory chow with added fat, principally corn oil, to produce a fat content of 16% or 23%). Homogenate of colon mucosa was fractionated by isopycnic banding on a linear sucrose gradient and the distribution of 5'-nucleotidase determined. Two main peaks were detected which we interpret as belonging to basolateral plasma membrane (BLPM) and brush borders. A high fat diet did not affect the relative sizes of the peaks or the median density of the BLPM peak but did significantly reduce the density of the brush border peak from 1.208 g/ml to 1.197 g/ml.


Brain cancer induces systemic immunosuppression through release of non-steroid soluble mediators.
Immunosuppression of unknown aetiology is a hallmark feature of glioblastoma and is characterized by decreased CD4 T-cell counts and downregulation of major histocompatibility complex class II expression on peripheral blood monocytes in patients. This immunosuppression is a critical barrier to the successful development of immunotherapies for glioblastoma. We recapitulated the immunosuppression observed in glioblastoma patients in the C57BL/6 mouse and investigated the aetiology of low CD4 T-cell counts. We determined that thymic involution was a hallmark feature of immunosuppression in three distinct models of brain cancer, including mice harbouring GL261 glioma, B16 melanoma, and in a spontaneous model of diffuse intrinsic pontine glioma. In addition to thymic involution, we determined that tumour growth in the brain induced significant splenic involution, reductions in peripheral T cells, reduced MHC II expression on blood leucocytes, and a modest increase in bone marrow resident CD4 T cells. Using parabiosis we report that thymic involution, declines in peripheral T-cell counts, and reduced major histocompatibility complex class II expression levels were mediated through circulating blood-derived factors. Conversely, T-cell sequestration in the bone marrow was not governed through circulating factors. Serum isolated from glioma-bearing mice potently inhibited proliferation and functions of T cells both in vitro and in vivo. Interestingly, the factor responsible for immunosuppression in serum is non-steroidal and of high molecular weight. Through further analysis of neurological disease models, we determined that the immunosuppression was not unique to cancer itself, but rather occurs in response to brain injury. Non-cancerous acute neurological insults also induced significant thymic involution and rendered serum immunosuppressive. Both thymic involution and serum-derived immunosuppression were reversible upon clearance of brain insults. These findings demonstrate that brain cancers cause multifaceted immunosuppression and pinpoint circulating factors as a target of intervention to restore immunity.


Anchoring biases affect repeated scores of thermal, moisture, tactile and comfort sensations in transient conditions.
In this study, we addressed potential biases which can occur when sensorial scores of temperature, wetness and discomfort are repeatedly reported, in transient exercise conditions. We pointed out that, when repeatedly reported, previous sensorial scores can be set by the participants as reference values and the subsequent score may be given based on the previous point of reference, the latter phenomenon leading to a bias which we defined as 'anchoring bias'. Indeed, the findings shown that subsequent sensorial scores are prone to anchoring biases and that the bias consisted in a systematically higher magnitude of sensation as compared to when reported a single time only. As such, the study allowed recognition, quantification and mitigation of the identified bias which can improve the methodological rigour of research studies involving assessments of sensorial data in transient conditions.


Alcohol use in emerging adults associated with lower rich-club connectivity and greater connectome network disorganization.
Emerging adulthood is a critical neurodevelopmental stage, with alcohol use during this period consistently associated with brain abnormalities and damage in anatomical structure and white matter integrity. However, it is less clear how alcohol use is associated with the brain's structural organization (i.e., white matter connections between anatomical regions). Recent connectome research has focused on rich-club regions, a collection of highly-interconnected hubs that are critical in brain communication and global network organization and disproportionately vulnerable to insults. For the first time, we examined alcohol use associations with structural rich-club and connectome organization in emerging adults (N = 66). Greater lifetime drinks and current monthly drinks were significantly associated with lower rich-club organization (rs =-0.38, ps < 0.003) and lower rich-club connectivity (rs <-0.34, ps < 0.007). Additionally, rich-club connectivity was significantly more negatively correlated with alcohol use than connectivity among non-rich-club regions (ps < 0.035). Examining overall structural organization, greater lifetime drinks and current monthly drinks were significantly associated with lower network density (i.e., lower network resilience; rs <-0.36, ps = 0.004). Additionally, greater lifetime drinks and current monthly drinks were significantly associated with higher network segregation (i.e., network's tendency to divide into subnetworks; rs >0.33, ps<0.008). Alcohol use was not significantly associated with network integration (i.e., network's efficiency in combining information across the brain; ps > 0.064). Results provide novel evidence that alcohol use is associated with decreased rich-club connectivity and structural network disorganization. Given that both are critical in global brain communication, these results highlight the importance of examining alcohol use and brain relationships in emerging adulthood.


The living arrangements of children of immigrants.
Children of immigrants are a rapidly growing part of the U.S. child population. Their health, development, educational attainment, and social and economic integration into the nation's life will play a defining role in the nation's future. Nancy Landale, Kevin Thomas, and Jennifer Van Hook explore the challenges facing immigrant families as they adapt to the United States, as well as their many strengths, most notably high levels of marriage and family commitment. The authors examine differences by country of origin in the human capital, legal status, and social resources of immigrant families and describe their varied living arrangements, focusing on children of Mexican, Southeast Asian, and black Caribbean origin. Problems such as poverty and discrimination may be offset for children to some extent by living, as many do, in a two-parent family. But the strong parental bonds that initially protect them erode as immigrant families spend more time in the United States and are swept up in the same social forces that are increasing single parenthood among American families. The nation, say the authors, should pay special heed to how this aspect of immigrants' Americanization heightens the vulnerability of their children. One risk factor for immigrant families is the migration itself, which sometimes separates parents from their children. Another is the mixed legal status of family members. Parents' unauthorized status can mire children in poverty and unstable living arrangements. Sometimes unauthorized parents are too fearful of deportation even to claim the public benefits for which their children qualify. A risk factor unique to refugees, such as Southeast Asian immigrants, is the death of family members from war or hardship in refugee camps. The authors conclude by discussing how U.S. immigration policies shape family circumstances and suggest ways to alter policies to strengthen immigrant families. Reducing poverty, they say, is essential. The United States has no explicit immigrant integration policy or programs, so policy makers must direct more attention and resources toward immigrant settlement, especially ensuring that children have access to the social safety net.


Cole function and conductance-based parasitic capacitance compensation for cerebral electrical bioimpedance measurements.
One of the most common measurement artifacts present in Electrical Bioimpedance Spectroscopy measurements (EBIS) comes from the capacitive leakage effect resulting from parasitic stray capacitances. This artifact produces a deviation in the measured impedance spectrum that is most noticeable at higher frequencies. The artifact taints the spectroscopy measurement increasing the difficulty of producing reliable EBIS measurements at high frequencies. In this work, an approach for removing such capacitive influence from the spectral measurement is presented making use of a novel method to estimate the value of the parasitic capacitance equivalent that causes the measurement artifact. The proposed method has been tested and validated theoretically and experimentally and it gives a more accurate estimation of the value of the parasitic capacitance than the previous methods. Once a reliable value of parasitic capacitance has been estimated the capacitive influence can be easily compensated in the EBIS measured data. Thus enabling analysis of EBIS data at higher frequencies, i.e. in the range of 300-500 kHz like measurements intended for cerebral monitoring, where the characteristic frequency is remarkably higher than EBIS measurements i.e. within the range 30 to 50 kHz, intended for body composition assessment.


Approaches to minimising the epidemiological impact of sources of systematic and random variation that may affect biochemistry assay data in UK Biobank.
Background: UK Biobank is a large prospective study that recruited 500,000 participants aged 40 to 69 years, between 2006-2010.The study has collected (and continues to collect) extensive phenotypic and genomic data about its participants. In order to enhance further the value of the UK Biobank resource, a wide range of biochemistry markers were measured in all participants with an available biological sample. Here, we describe the approaches UK Biobank has taken to minimise error related to sample collection, processing, retrieval and assay measurement. Methods: During routine quality control checks, the laboratory team observed that some assay results were lower than expected for samples acquired during certain time periods. Analyses were undertaken to identify and correct for the unexpected dilution identified during sample processing, and for expected error caused by laboratory drift of assay results. Results: The vast majority (92%) of biochemistry serum assay results were assessed to be not materially affected by dilution, with an estimated difference in concentration of less than 1% (i.e. either lower or higher) than that expected if the sample were unaffected; 8.3% were estimated to be diluted by up to 10%; very few samples appeared to be diluted more than this. Biomarkers measured in urine (creatinine, microalbumin, sodium, potassium) and red blood cells (HbA1c) were not affected. In order to correct for laboratory variation over the assay period, all assay results were adjusted for date of assay, with the exception of those that had a high biological coefficient of variation or evident seasonal variability: vitamin D, lipoprotein (a), gamma glutamyltransferase, C-reactive protein and rheumatoid factor. Conclusions: Rigorous approaches related to sample collection, processing, retrieval, assay measurement and data analysis have been taken to mitigate the impact of both systematic and random variation in epidemiological analyses that use the biochemistry assay data in UK Biobank.


The safety helmet detection technology and its application to the surveillance system.
The Automatic Teller Machine (ATM) plays an important role in the modem economy. It provides a fast and convenient way to process transactions between banks and their customers. Unfortunately, it also provides a convenient way for criminals to get illegal money or use stolen ATM cards to extract money from their victims' accounts. For safety reasons, each ATM has a surveillance system to record customer's face information. However, when criminals use an ATM to withdraw money illegally, they usually hide their faces with something (in Taiwan, criminals usually use safety helmets to block their faces) to avoid the surveillance system recording their face information, which decreases the efficiency of the surveillance system. In this paper, we propose a circle/circular arc detection method based upon the modified Hough transform, and apply it to the detection of safety helmets for the surveillance system of ATMs. Since the safety helmet location will be within the set of the obtainable circles/circular arcs (if any exist), we use geometric features to verify if any safety helmet exists in the set. The proposed method can be used to help the surveillance systems record a customer's face information more precisely. If customers wear safety helmets to block their faces, the system can send a message to remind them to take off their helmets. Besides this, the method can be applied to the surveillance systems of banks by providing an early warning safeguard when any "customer" or "intruder" uses a safety helmet to avoid his/her face information from being recorded by the surveillance system. This will make the surveillance system more useful. Real images are used to analyze the performance of the proposed method.


The effect of physical fatigue on oscillatory dynamics of the sensorimotor cortex.
While physical fatigue is known to arise in part from supraspinal mechanisms within the brain, exactly how brain activity is modulated during fatigue is not well understood. Therefore, this study examined how typical neural oscillatory responses to voluntary muscle contractions were affected by fatigue. Eleven healthy adults (age 27 ± 4 years) completed two experimental sessions in a randomized crossover design. Both sessions first assessed baseline maximal voluntary isometric wrist-flexion force (MVFb ). Participants then performed an identical series of fourteen test contractions (2 × 100%MVFb , 10 × 40%MVFb , 2 × 100%MVFb ) both before and after one of two interventions: forty 12-s contractions at 55%MVFb (fatigue intervention) or 5%MVFb (control intervention). Magnetoencephalography (MEG) was used to characterize both the movement-related mu and beta decrease (MRMD and MRBD) and the post-movement beta rebound (PMBR) within the contralateral sensorimotor cortex during the 40%MVFb test contractions, while the 100%MVFb test contractions were used to monitor physical fatigue. The fatigue intervention induced a substantial physical fatigue that endured throughout the post-intervention measurements (28.9-29.5% decrease in MVF, P < 0.001). Fatigue had a significant effect on both PMBR (anova, session × time-point interaction: P = 0.018) and MRBD (P = 0.021): the magnitude of PMBR increased following the fatigue but not the control interventions, whereas MRBD was decreased post-control but not post-fatigue. Mu oscillations were unchanged throughout both sessions. Physical fatigue resulted in an increased PMBR, and offset attenuations in MRBD associated with task habituation.


Investigating Biological Control Agents for Controlling Invasive Populations of the Mealybug Pseudococcus comstocki in France.
Pseudococcus comstocki (Hemiptera: Pseudococcidae) is a mealybug species native to Eastern Asia and present as an invasive pest in northern Italy and southern France since the start of the century. It infests apple and pear trees, grapevines and some ornamental trees. Biocontrol programmes against this pest proved successful in central Asia and North America in the second half of the 20th century. In this study, we investigated possible biocontrol agents against P. comstocki, with the aim of developing a biocontrol programme in France. We carried out systematic DNA-barcoding at each step in the search for a specialist parasitoid. First we characterised the French target populations of P. comstocki. We then identified the parasitoids attacking P. comstocki in France. Finally, we searched for foreign mealybug populations identified a priori as P. comstocki and surveyed their hymenopteran parasitoids. Three mealybug species (P. comstocki, P. viburni and P. cryptus) were identified during the survey, together with at least 16 different parasitoid taxa. We selected candidate biological control agent populations for use against P. comstocki in France, from the species Allotropa burrelli (Hymenoptera: Platygastridae) and Acerophagus malinus (Hymenoptera: Encyrtidae). The coupling of molecular and morphological characterisation for both pests and natural enemies facilitated the programme development and the rejection of unsuitable or generalist parasitoids.


The use of a pectoralis major flap to improve internal mammary vessels exposure and reduce contour deformity in microvascular free flap breast reconstruction.
Internal mammary vessels are commonly used in microvascular breast reconstruction. Most surgeons resect a portion of the rib to expose these vessels. This resection can lead to contour abnormalities in the chest wall that are difficult to correct. In addition, exposure of these vessels deep in the wound can be problematic. The purpose of this study was to evaluate our experience with a pectoralis major flap designed to improve exposure and fill in the defect created by rib resection. All consecutive patients who underwent autologous breast reconstruction using the internal mammary vessels as recipient vessels between 2000 and 2005 were identified. All procedures were performed by a single surgeon. In each case, a superiorly based flap within the pectoralis major muscle spanning the bottom of the second rib to the top of the fourth costal cartilage was raised. This "L"-shaped flap was reflected and a portion of the third rib cartilage was excised. At the conclusion of the microsurgical anastomosis the pectoralis major flap was repaired and used to cover the defect created by rib resection. Patient demographic, operative details, and postoperative complications were obtained from a prospectively maintained clinical database. Deformity around sternal border was evaluated from the patients' photographs. There were 99 autologous reconstructions in 90 patients. In 71 cases the internal mammary artery/vein were used as recipient vessels. There were no cases of microvascular thrombosis or flap loss. A portion of the third rib was excised in all patients who underwent microsurgical anastomoses to the internal mammary vessels. In 3 patients a portion of both the second and third ribs was removed because of branching of the internal mammary vein proximal to the level of the third rib. A contour deformity was noted in 4 patients (4.4%) after a mean follow-up of 27 months. Of the 4 patients with contour deformity, 2 had a portion of both the third and second costal cartilage removed because of venous branching above the level of the third rib cartilage. The modified pectoralis major L-shaped flap is a useful technique for safe and clear exposure of the internal mammary vessels. In addition, the use of this flap to cover the segment of resected rib cartilage can decrease the contour deformities associated with rib resection as compared with reported rates in the literature. Resection of multiple rib segments, though unavoidable at times because of anatomic considerations, may be associated with an increased rate of postoperative contour deformities.


Borderline personality disorder criteria associated with prospectively observed suicidal behavior.
The authors prospectively examined associations between each DSM-IV borderline personality disorder criterion and suicidal behaviors. Borderline personality disorder diagnosis and criteria, major depressive disorder, substance use disorders, and history of childhood sexual abuse were assessed with semistructured interviews. Participants (N=621) were followed for 2 years with repeated structured evaluations that included assessments of suicidality. With the self-injury criterion excluded, the borderline personality disorder criteria of affective instability, identity disturbance, and impulsivity significantly predicted suicidal behaviors. Only affective instability and childhood sexual abuse were significantly associated with suicide attempts (i.e., behavior with some intent to die). Affective instability is the borderline personality disorder criterion (excluding self-injury) most strongly associated with suicidal behaviors. Since major depressive disorder did not significantly predict suicidal behaviors, the reactivity associated with affective instability (more so than negative mood states) appears to be a critical element in predicting suicidal behaviors.


A Systematic Review of Validity Evidence for the Fundamentals of Laparoscopic Surgery Examination in Gynecology.
The Fundamentals in Laparoscopic Surgery (FLS) examination is designed to test laparoscopic surgery skills. Our aim for this systematic review was to examine validity evidence supporting or refuting the FLS examination specifically as a high-stakes summative assessment tool in gynecology. The data sources were PubMed, MEDLINE, Embase, and Scopus. The study eligibility criterion was the subject of the FLS examination as an assessment tool in gynecology. We developed a data extraction tool and assigned articles for screening and extraction to all authors, who then abstracted data independently. Conflicts that arose during the extraction process were resolved by consensus. We organized validity evidence for the cognitive and manual skills portions on the basis of the categories of current validation standards. From 1971 citations identified, 9 studies were included, involving 319 participants. For the cognitive portion of the test, the results were mixed in 5 studies in relationships with the other variables category. For the manual portion of the test, most of the studies focused on the relationships with other variables evidence with mixed findings. The concerning findings in the manual skills portion included the lack of transferability of skills to the operating room, limited mixed evidence for improvement in operating room performance, and worse performance by obstetrics and gynecology surgeons compared with other specialties. We did not find supportive content-based, response process, or consequential evidence in either the cognitive or manual skills portion of the test. Validity evidence for the FLS examination was either mixed, as it pertained to relationships with other variables, or lacking in other important evidence categories. Further evidence is required to justify the use of the FLS examination scores as a high-stakes summative assessment.


A Thermotolerant Variant of Rubisco Activase From a Wild Relative Improves Growth and Seed Yield in Rice Under Heat Stress.
Genes encoding thermostable variants of the photosynthesis heat-labile protein Rubisco activase (Rca) from a wild relative Oryza australiensis were overexpressed in domesticated rice (Oryza sativa). Proteomics was used to quantify the abundance of O. australiensis Rca (Rca-Oa) in the resulting plants. Plants were grown to maturity in growth rooms and from early tillering until immediately prior to anthesis, they were exposed to daytime maximum temperatures of 28, 40, and 45°C and constant night temperatures of 22°C. Non-destructive measurements of leaf elongation and photosynthesis were used to compare the null segregant with a transfected line in which 19% of its total Rca content was the recombinant O. australiensis Rca (T-Oa-19). Height, fresh mass, panicle number, seed set, and seed number were measured at final harvest. Traits at maturity after heat stress at 45°C correlated strongly with recombinant protein abundance. Seed number was far the most responsive trait to an increase in Rca-Oa abundance, improving by up to 150%. Leaf elongation rates (LER) and tiller number were significantly greater in the transformed plants in the first two weeks of exposure to 45°C but tiller numbers later became equal in the two genotypes. Gas exchange measurements showed that T-Oa-19 had faster light induction of photosynthesis but not significantly higher CO2 assimilation rates, indicating that the carbon gain that resulted in large yield improvement after growth at 45°C was not strongly correlated with an instantaneous measurement of steady-state photosynthesis. When plants were grown at 40°C daytime maximum, there was no improvement in the final biomass, panicle or seed number when compared with 28°C, indicating that the threshold for heat damage and beneficial effects of the thermostable Rca recombinant protein was between 40 and 45°C, which corresponded to leaf temperatures in the range 38-42°C. The results suggest that the thermotolerant form of Rca from O. australiensis was sufficient to enhance carbohydrate accumulation and storage by rice over the life of the plant, dramatically improving yields after exposure to heat throughout the vegetative phase.


Sleep problems in children.
Sleep complaints and sleep disorders are common during childhood and adolescence. The impact of not getting enough sleep may affect children's' physical health as well emotional, cognitive and social development. Insomnia, sleep-disordered breathing, parasomnias and sleep disturbances associated with medical and psychiatric disorders are some of the commonly encountered sleep disorders in this age group. Changes in sleep architecture and the amount of sleep requirement associated with each stage of development should be considered during an evaluation of sleep disorders in children. Behavioral treatments should be used initially wherever possible especially considering that most pharmacologic agents used to treat pediatric sleep disorders are off-label. In this review we address the most common sleep problems in children/adolescents as they relate to prevalence, presentation and symptoms, evaluation and management.


Use of green rooibos (Aspalathus linearis) extract and water-soluble nanomicelles of green rooibos extract encapsulated with ascorbic acid for enhanced aspalathin content in ready-to-drink iced teas.
Heat-induced changes in aspalathin, iso-orientin, and orientin content of ready-to-drink (RTD) green rooibos iced tea formulations were investigated. An organic-solvent-based aspalathin-enriched extract prepared from green rooibos was used "as-is" or encapsulated with ascorbic acid in a water-soluble nanomicelle-based carrier system. The common iced tea ingredients, ascorbic acid, and/or citric acid were added to the iced tea containing green rooibos extract. Only citric acid was added to the iced tea containing the nanomicelles. Heat treatments consisted of pasteurization (93 °C/5 min and 93 °C/30 min), normal-temperature sterilization (NTS; 121 °C/15 min), and high-temperature sterilization (HTS; 135 °C/4 min). Pasteurization had little or no effect on the flavonoid content. NTS and HTS induced significant losses in the flavonoids. The addition of citric and ascorbic acids improved the stability of the flavonoids, but encapsulation of green rooibos extract with ascorbic acid in nanomicelles did not offer additional stability. The only benefit of using the water-soluble nanomicelles was the improved clarity of the RTD product. Iso-orientin and orientin contents were substantially less affected than aspalathin by the heat treatments, partially because of conversion of aspalathin to these flavones, which countered losses. 5-Hydroxymethylfurfural (HMF), a known dehydration product of hexoses under mild acidic conditions and also a degradation product of ascorbic acid, was observed in formulations containing citric and/or ascorbic acids.


Efficacy of deguelin and silibinin on benzo(a)pyrene-induced lung tumorigenesis in A/J mice.
We evaluated deguelin and silibinin in A/J mice treated with the tobacco-specific carcinogen benzo(a)pyrene (BP) for their ability to inhibit pulmonary adenoma formation and growth. Animals were treated with either deguelin (5.0 or 10.0 mg/kg body weight, by gavage) or silibinin at doses of 0.05% and 0.1% in the diet, approximately 10 days before a single intraperitoneal dose of BP. We found that oral administration of deguelin reduced tumor multiplicity by 56% and tumor load by 78%, whereas silibinin treatment at doses of 0.05% and 0.1% in the diet did not show any significant efficacy on either tumor multiplicity or tumor load. The result indicates that deguelin significantly inhibits pulmonary adenoma formation and growth in A/J mice. Finding new and effective agents that can prevent lung cancer is urgently needed because cancer of the lungs remains the principal cause of cancer deaths in the United States and because effective chemoprevention of this cancer type remains elusive. Thus, deguelin appears to be a promising new preventive agent for lung cancer and may be considered for further studies in other animal models and in clinical trials.


'Lingering' opiate deaths? Concentration of opiates in medulla and femoral blood.
'Lingering death' cases occur when the circumstances of death indicate an opiate overdose, but measured opiate blood levels are only in the therapeutic range; death results from cardiac and respiratory depression. This study examined the relative concentration of opiates in femoral blood and in the medulla oblongata (sites for cardiac and respiratory control) from 41 cases to determine whether a difference in opiate concentration might explain lingering deaths. Opiates from blood and medulla were analyzed using GC-EI-MS in selective ion monitoring mode. Results were correlated with gross and microscopic findings of the lungs and with cause and manner of death. Opiate concentrations for morphine, codeine and 6-acetylmorphine (6-AM) were higher in the medulla than in blood. The brain: blood ratio for the analytes demonstrated an increasing ratio from morphine, to codeine, to 6-AM (1.42, 2.48 and 4.86), which corresponds to the relative lipophilicity of these analytes. The average right and left lung weights were 762 and 668 g, respectively. Histologic examination showed edema, and/or polarizable microemboli, acute bronchopneumonia and acute bronchitis. The preferential distribution of opiates to medulla suggests that lingering opiate deaths may be explained, at least in part, because of higher relative concentrations of drug in brain, compared with femoral blood.


Regulation of mitotic spindle formation by the RhoA guanine nucleotide exchange factor ARHGEF10.
The Dbl family guanine nucleotide exchange factor ARHGEF10 was originally identified as the product of the gene associated with slowed nerve-conduction velocities of peripheral nerves. However, the function of ARHGEF10 in mammalian cells is totally unknown at a molecular level. ARHGEF10 contains no distinctive functional domains except for tandem Dbl homology-pleckstrin homology and putative transmembrane domains. Here we show that RhoA is a substrate for ARHGEF10. In both G1/S and M phases, ARHGEF10 was localized in the centrosome in adenocarcinoma HeLa cells. Furthermore, RNA interference-based knockdown of ARHGEF10 resulted in multipolar spindle formation in M phase. Each spindle pole seems to contain a centrosome consisting of two centrioles and the pericentriolar material. Downregulation of RhoA elicited similar phenotypes, and aberrant mitotic spindle formation following ARHGEF10 knockdown was rescued by ectopic expression of constitutively activated RhoA. Multinucleated cells were not increased upon ARHGEF10 knockdown in contrast to treatment with Y-27632, a specific pharmacological inhibitor for the RhoA effector kinase ROCK, which induced not only multipolar spindle formation, but also multinucleation. Therefore, unregulated centrosome duplication rather than aberration in cytokinesis may be responsible for ARHGEF10 knockdown-dependent multipolar spindle formation. We further isolated the kinesin-like motor protein KIF3B as a binding partner of ARHGEF10. Knockdown of KIF3B again caused multipolar spindle phenotypes. The supernumerary centrosome phenotype was also observed in S phase-arrested osteosarcoma U2OS cells when the expression of ARHGEF10, RhoA or KIF3B was abrogated by RNA interference. Collectively, our results suggest that a novel RhoA-dependent signaling pathway under the control of ARHGEF10 has a pivotal role in the regulation of the cell division cycle. This pathway is not involved in the regulation of cytokinesis, but instead may regulate centrosome duplication. The kinesin-like motor protein KIF3B may modulate the ARHGEF10-RhoA pathway through the binding to ARHGEF10.


Loading of DNA-binding factors to an erythroid enhancer.
The HS-40 enhancer is the major cis-acting regulatory element responsible for the developmental stage- and erythroid lineage-specific expression of the human alpha-like globin genes, the embryonic zeta and the adult alpha2/alpha/1. A model has been proposed in which competitive factor binding at one of the HS-40 motifs, 3'-NA, modulates the capability of HS-40 to activate the embryonic zeta-globin promoter. Furthermore, this modulation was thought to be mediated through configurational changes of the HS-40 enhanceosome during development. In this study, we have further investigated the molecular basis of this model. First, human erythroid K562 cells stably integrated with various HS-40 mutants cis linked to a human alpha-globin promoter-growth hormone hybrid gene were analyzed by genomic footprinting and expression analysis. By the assay, we demonstrate that factors bound at different motifs of HS-40 indeed act in concert to build a fully functional enhanceosome. Thus, modification of factor binding at a single motif could drastically change the configuration and function of the HS-40 enhanceosome. Second, a specific 1-bp, GC-->TA mutation in the 3'-NA motif of HS-40, 3'-NA(II), has been shown previously to cause significant derepression of the embryonic zeta-globin promoter activity in erythroid cells. This derepression was hypothesized to be regulated through competitive binding of different nuclear factors, in particular AP1 and NF-E2, to the 3'-NA motif. By gel mobility shift and transient cotransfection assays, we now show that 3'-NA(II) mutation completely abolishes the binding of small MafK homodimer. Surprisingly, NF-E2 as well as AP1 can still bind to the 3'-NA(II) sequence. The association constants of both NF-E2 and AP1 are similar to their interactions with the wild-type 3'-NA motif. However, the 3'-NA(II) mutation causes an approximately twofold reduction of the binding affinity of NF-E2 factor to the 3'-NA motif. This reduction of affinity could be accounted for by a twofold-higher rate of dissociation of the NF-E2-3'-NA(II) complex. Finally, we show by chromatin immunoprecipitation experiments that only binding of NF-E2, not AP1, could be detected in vivo in K562 cells around the HS-40 region. These data exclude a role for AP1 in the developmental regulation of the human alpha-globin locus via the 3'-NA motif of HS-40 in embryonic/fetal erythroid cells. Furthermore, extrapolation of the in vitro binding studies suggests that factors other than NF-E2, such as the small Maf homodimers, are likely involved in the regulation of the HS-40 function in vivo.


A Prospective Safety Trial of Atorvastatin Treatment to Assess Rebleeding after Spontaneous Intracerebral Hemorrhage: A Serial MRI Investigation.
This study was designed to determine any rebleeding after atorvastatin treatment following spontaneous intracerebral hemorrhage (ICH) in a prospective safety trial. Atorvastatin (80 mg/day) therapy was initiated in 6 patients with primary ICH with admission Glasgow Coma Score (GCS) >5 within 24 hours of ictus and continued for 7 days, with the dose tapered and treatment terminated over the next 5 days. Patients were studied longitudinally by multiparametric magnetic resonance imaging (MRI) at three time points: acute (3 to 5 days), subacute (4 to 6 weeks) and chronic (3 to 4 months). Imaging sequences included T1, T2-weighted imaging (T2WI), diffusion tensor imaging (DTI) and contrast-enhanced MRI measures of cerebral perfusion, blood volume and blood-brain barrier (BBB) permeability. Susceptibility weighted imaging (SWI) was used to identify primary ICH and to check for secondary rebleeding. Final outcome was assessed using Glasgow Outcome Score (GOS) at 3-4 months. Mean admission GCS was 13.2±4.0 and mean GOS at 3 months was 4.5±0.6. Hemorrhagic lesions were segmented into core and rim areas. Mean lesion volumes decreased significantly between the acute and chronic study time points (p=0.008). Average ipsilateral hemispheric tissue loss at 3 to 4 months was 11.4±4.6 cm3. MRI showed acutely reduced CBF (p=0.004) and CBV (p=0.002) in the rim, followed by steady normalization. Apparent diffusion coefficient of water (ADC) in the rim demonstrated no alterations at any of the time points (p>0.2). The T2 values were significantly elevated in the rim acutely (p=0.02), but later returned to baseline. The ICH core showed sustained low CBF and CBV values concurrent with a small reduction in ADC acutely, but significant ADC elevation at the end suggestive of irreversible injury. Despite the presence of a small, probably permanent, cerebral lesion in the ICH core, no patients exhibited post-treatment rebleeding. These data suggest that larger, Phase 2 trials are warranted to establish long term clinical safety of atorvastatin in spontaneous ICH.


Lack of FTSH4 Protease Affects Protein Carbonylation, Mitochondrial Morphology, and Phospholipid Content in Mitochondria of Arabidopsis: New Insights into a Complex Interplay.
FTSH4 is one of the inner membrane-embedded ATP-dependent metalloproteases in mitochondria of Arabidopsis (Arabidopsis thaliana). In mutants impaired to express FTSH4, carbonylated proteins accumulated and leaf morphology was altered when grown under a short-day photoperiod, at 22°C, and a long-day photoperiod, at 30°C. To provide better insight into the function of FTSH4, we compared the mitochondrial proteomes and oxyproteomes of two ftsh4 mutants and wild-type plants grown under conditions inducing the phenotypic alterations. Numerous proteins from various submitochondrial compartments were observed to be carbonylated in the ftsh4 mutants, indicating a widespread oxidative stress. One of the reasons for the accumulation of carbonylated proteins in ftsh4 was the limited ATP-dependent proteolytic capacity of ftsh4 mitochondria, arising from insufficient ATP amount, probably as a result of an impaired oxidative phosphorylation (OXPHOS), especially complex V. In ftsh4, we further observed giant, spherical mitochondria coexisting among normal ones. Both effects, the increased number of abnormal mitochondria and the decreased stability/activity of the OXPHOS complexes, were probably caused by the lower amount of the mitochondrial membrane phospholipid cardiolipin. We postulate that the reduced cardiolipin content in ftsh4 mitochondria leads to perturbations within the OXPHOS complexes, generating more reactive oxygen species and less ATP, and to the deregulation of mitochondrial dynamics, causing in consequence the accumulation of oxidative damage.


Protocol for a statewide randomized controlled trial to compare three training models for implementing an evidence-based treatment.
Evidence-based treatments (EBTs) are available for treating childhood behavioral health challenges. Despite EBTs' potential to help children and families, they have primarily remained in university settings. Little empirical evidence exists regarding how specific, commonly used training and quality control models are effective in changing practice, achieving full implementation, and supporting positive client outcomes. This study (NIMH RO1 MH095750; ClinicalTrials.gov Identifier: NCT02543359), which is currently in progress, will evaluate the effectiveness of three training models (Learning Collaborative (LC), Cascading Model (CM), and Distance Education (DE)) to implement a well-established EBT , Parent-Child Interaction Therapy, in real-world, community settings. The three models differ in their costs, skill training, quality control methods, and capacity to address broader implementation challenges. The project is guided by three specific aims: (1) to build knowledge about training outcomes, (2) to build knowledge about implementation outcomes, and (3) to test the differential impact of training clinicians using LC, CM, and DE models on key client outcomes. Fifty (50) licensed psychiatric clinics across Pennsylvania were randomized to one of the three training conditions: (1) LC, (2) CM, or (3) DE. The impact of training on practice skills (clinician level) and implementation/sustainment outcomes (clinic level) are being evaluated at four timepoints coinciding with the training schedule: baseline, 6 (mid), 12 (post), and 24 months (1 year follow-up). Immediately after training begins, parent-child dyads (client level) are recruited from the caseloads of participating clinicians. Client outcomes are being assessed at four timepoints (pre-treatment, 1, 6, and 12 months after the pre-treatment). This proposal builds on an ongoing initiative to implement an EBT statewide. A team of diverse stakeholders including state policy makers, payers, consumers, service providers, and academics from different, but complementary areas (e.g., public health, social work, psychiatry), has been assembled to guide the research plan by incorporating input from multidimensional perspective. ClinicalTrials.gov: NCT02543359.


In situ drug-receptor binding kinetics in single cells: a quantitative label-free study of anti-tumor drug resistance.
Many drugs are effective in the early stage of treatment, but patients develop drug resistance after a certain period of treatment, causing failure of the therapy. An important example is Herceptin, a popular monoclonal antibody drug for breast cancer by specifically targeting human epidermal growth factor receptor 2 (Her2). Here we demonstrate a quantitative binding kinetics analysis of drug-target interactions to investigate the molecular scale origin of drug resistance. Using a surface plasmon resonance imaging, we measured the in situ Herceptin-Her2 binding kinetics in single intact cancer cells for the first time, and observed significantly weakened Herceptin-Her2 interactions in Herceptin-resistant cells, compared to those in Herceptin-sensitive cells. We further showed that the steric hindrance of Mucin-4, a membrane protein, was responsible for the altered drug-receptor binding. This effect of a third molecule on drug-receptor interactions cannot be studied using traditional purified protein methods, demonstrating the importance of the present intact cell-based binding kinetics analysis.


Time trends in mortality in forestry and construction workers in Finland 1970-85 and impact of adjustment for socioeconomic variables.
This study aimed firstly to describe the development of cause-specific mortality in forestry workers, farmer/forestry workers, and skilled and semiskilled construction workers between 1970 and 1985 in Finland, and to compare this with mortality in the total working male population. The second aim was to evaluate how well the cause-specific mortality differences between the occupations could be explained by differences in socioeconomic status, marital status, or in the region of residence. This is a follow up study based on the 1970, 1975, and 1980 census records in Finland linked with all death certificates for 1971-75, 1976-80, and 1981-85 respectively. Log-linear regression analysis was used. All economically active men in Finland aged between 35 and 64 years in 1971-85 were studied. The number of person-years in the period totals about 10 million. Semiskilled construction workers had the highest mortality rate almost independent of the cause of death. The mortality of forestry workers was the second highest. Compared with the reference population, however, the differences were small with regard to neoplasms and cardiovascular diseases. With regard to other diseases, only the mortality of semiskilled workers was fairly high. Differences in mortality as a result of accidents were the highest. Both suicide and accidental death rates were high in semi-skilled construction workers and forestry workers. During the study total mortality fell by about 30% but mortality differences between groups did not decline. The high mortality of forestry and semiskilled construction workers is partly explained by socioeconomic factors such as marital status and housing conditions. These factors do not, however, explain the high suicide and accident mortality rates of forestry workers or semiskilled construction workers. More research is needed to explain these findings.


LTQ-XL mass spectrometry proteome analysis expands the Pseudomonas aeruginosa AmpR regulon to include cyclic di-GMP phosphodiesterases and phosphoproteins, and identifies novel open reading frames.
Pseudomonas aeruginosa is well known for its antibiotic resistance and intricate regulatory network, contributing to its success as an opportunistic pathogen. This study is an extension of our transcriptomic analyses (microarray and RNA-Seq) to understand the global changes in PAO1 upon deleting a gene encoding a transcriptional regulator AmpR, in the presence and absence of β-lactam antibiotic. This study was performed under identical conditions to explore the proteome profile of the ampR deletion mutant (PAOΔampR) using LTQ-XL mass spectrometry. The proteomic data identified ~53% of total PAO1 proteins and expanded the master regulatory role of AmpR in determining antibiotic resistance and multiple virulence phenotypes in P. aeruginosa. AmpR proteome analysis identified 853 AmpR-dependent proteins, which include 102 transcriptional regulators and 21 two-component system proteins. AmpR also regulates cyclic di-GMP phosphodiesterases (PA4367, PA4969, PA4781) possibly affecting major virulence systems. Phosphoproteome analysis also suggests a significant role for AmpR in Ser, Thr and Tyr phosphorylation. These novel mechanisms of gene regulation were previously not associated with AmpR. The proteome analysis also identified many unannotated and misannotated ORFs in the P. aeruginosa genome. Thus, our data sheds light on important virulence regulatory pathways that can potentially be exploited to deal with P. aeruginosa infections. The AmpR proteome data not only confirmed the role of AmpR in virulence and resistance to multiple antibiotics, but also expanded the perimeter of AmpR regulon. The data presented here points to the role of AmpR in regulating cyclic di-GMP levels and phosphorylation of Ser, Thr and Tyr, adding another dimension to the regulatory functions of AmpR. We also identify some previously unannotated/misannotated ORFs in the P. aeruginosa genome, indicating the limitations of existing ORF analyses software. This study will contribute towards understanding complex genetic organization of P. aeruginosa. Whole genome proteomic picture of regulators at higher nodal positions in the regulatory network will not only help us link various virulence phenotypes but also design novel therapeutic strategies.


Kaposi's sarcoma cells: long-term culture with growth factor from retrovirus-infected CD4+ T cells.
Studies of the biology and pathogenesis of Kaposi's sarcoma (KS) have been hampered by the inability to maintain long-term cultures of KS cells in vitro. In this study AIDS-KS-derived cells with characteristic spindle-like morphology were cultured with a growth factor (or factors) released by CD4+ T lymphocytes infected with human T-lymphotropic virus type I or II (HTLV-I or HTLV-II) or with human immunodeficiency virus type 1 or 2 (HIV-1 or HIV-2). Medium conditioned by HTLV-II-infected, transformed lines of T cells (HTLV-II CM) contained large amounts of this growth activity and also supported the temporary growth of normal vascular endothelial cells, but not fibroblasts. Interleukin-1 and tumor necrosis factor-alpha stimulated the growth of the KS-derived cells, but the growth was only transient and these could be distinguished from that in HTLV-II CM. Other known endothelial cell growth promoting factors, such as acidic and basic fibroblast growth factors and epidermal growth factor, did not support the long-term growth of the AIDS-KS cells. The factor released by CD4+ T cells infected with human retroviruses should prove useful in studies of the pathogenesis of KS.


The interrelationship between grip work, self-perceived fatigue and pre-frailty in community-dwelling octogenarians.
Low grip work and high feelings of self-perceived fatigue could be an early characteristic of decline in reserve capacity, which comes to full expression as physical frailty in a later stage. When grip work and self-perceived fatigue can be identified as characteristics differentiating between robustness and pre-frailty it might allow to identify pre-frailty earlier. Therefore, this study aimed to investigate whether the combination of grip work and self-perceived fatigue is related to pre-frailty in well-functioning older adults aged 80 and over. Four-hundred and five community-dwelling older adults aged 80 and over (214 robust and 191 pre-frail) were assessed for muscle endurance (grip Work corrected for body weight (GW_bw)), self-perceived fatigue (MFI-20) and frailty state (Fried Frailty Index, FFI). A Capacity to Perceived Vitality ratio (CPV) was calculated by dividing GW_bw by the MFI-20 scores. ANCOVA analysis (corrected for age and gender) was used to compare robust and pre-frail older adults, and binary logistic regressions were applied to analyze the relationship between CPV and pre-frailty status. Pre-frail older adults who scored negative on the exhaustion item of the FFI still showed significantly lower GW (p < 0.001), CPV ratios (p < 0.001) and higher self-perceived fatigue (p < 0.05) compared to the robust ones. The likelihood for pre-frailty related significantly to higher age, being men and lower CPV ratios. In women, every unit increase in CPV ratio decreased the likelihood for pre-frailty by 78% (OR 0.22; 95% CI: 0.11-0.44), for men this effect was less strong (34%, OR 0.66; 95% CI: 0.47-0.93). Pre-frail community-dwelling persons aged 80 years and over without clinical signs of exhaustion on the FFI still experience significantly higher fatigue levels (lower Grip Work, higher self-perceived fatigue and lower CPV levels) compared to robust ones. CPV ratio could therefore be a good tool to identify subclinical fatigue in the context of physical (pre-)frailty.


Influence of atrial fibrillation on plasma von willebrand factor, soluble E-selectin, and N-terminal pro B-type natriuretic peptide levels in systolic heart failure.
Endothelial dysfunction is present in patients with heart failure (HF) due to left ventricular systolic dysfunction, as well as in patients with atrial fibrillation (AF) who have normal cardiac function. It is unknown whether AF influences the degree of endothelial dysfunction in patients with systolic HF. We measured levels of plasma von Willebrand factor (vWF) and E-selectin (as indexes of endothelial damage/dysfunction and endothelial activation, respectively; both enzyme-linked immunosorbent assay) in patients with AF and HF (AF-HF), who were compared to patients with sinus rhythm and HF (SR-HF), as well as in age-matched, healthy, control subjects. We also assessed the relationship of vWF and E-selectin to plasma N-terminal pro B-type natriuretic peptide (NTpro-BNP), a marker for HF severity and prognosis. One hundred ninety patients (73% men; mean age, 69.0 +/- 10.1 years [+/- SD]) with systolic HF were studied, who were compared to 117 healthy control subjects: 52 subjects (27%) were in AF, while 138 subjects (73%) were in sinus rhythm. AF-HF patients were older than SR-HF patients (p = 0.046), but left ventricular ejection fraction and New York Heart Association class were similar. There were significant differences in NT-proBNP (p < 0.0001) and plasma vWF (p = 0.003) between patients and control subjects. On Tukey post hoc analysis, AF-HF patients had significantly increased NT-proBNP (p < 0.001) and vWF (p = 0.0183) but not E-selectin (p = 0.071) levels when compared to SR-HF patients. On multivariate analysis, the presence of AF was related to plasma vWF levels (p = 0.018). Plasma vWF was also significantly correlated with NT-proBNP levels (Spearman r = 0.139; p = 0.017). There is evidence of greater endothelial damage/dysfunction in AF-HF patients when compared to SR-HF patients. The clinical significance of this is unclear but may have prognostic value.


Plasmon-plasmon hybridization and bandwidth enhancement in nanostructured graphene.
Graphene plasmonic structures with long-range layering periodicity are presented. Resonance energy scaling with the number of graphene layers involved in plasmonic excitation allows these structures to support multiple plasmonic modes that couple and hybridize due to their physical proximity. Hybridized states exhibit bandwidth enhancements of 100-200% compared to unhybridized modes, and resonance energies deviate from what is usually observed in coupled plasmonic systems. Origins of this behavior are discussed, and experimental observations are computationally modeled. This work is a precursor and template for the study of plasmonic hybridization in other two-dimensional material systems with layering periodicity.


Pharmacological inhibition of syntenin PDZ2 domain impairs breast cancer cell activities and exosome loading with syndecan and EpCAM cargo.
Exosomes support cell-to-cell communication in physiology and disease, including cancer. We currently lack tools, such as small chemicals, capable of modifying exosome composition and activity in a specific manner. Building on our previous understanding of how syntenin, and its PDZ partner syndecan (SDC), impact on exosome composition we optimized a small chemical compound targeting the PDZ2 domain of syntenin. In vitro , in tests on MCF-7 breast carcinoma cells, this compound is non-toxic and impairs cell proliferation, migration and primary sphere formation. It does not affect the size or the number of secreted particles, yet it decreases the amounts of exosomal syntenin, ALIX and SDC4 while leaving other exosomal markers unaffected. Interestingly, it also blocks the sorting of EpCAM, a bona fide target used for carcinoma exosome immunocapture. Our study highlights the first characterization of a small pharmacological inhibitor of the syntenin-exosomal pathway, of potential interest for exosome research and oncology.


Principal component analysis applied to Fourier transform infrared spectroscopy for the design of calibration sets for glycerol prediction models in wine and for the detection and classification of outlier samples.
Principal component analysis (PCA) was used to identify the main sources of variation in the Fourier transform infrared (FT-IR) spectra of 329 wines of various styles. The FT-IR spectra were gathered using a specialized WineScan instrument. The main sources of variation included the reducing sugar and alcohol content of the samples, as well as the stage of fermentation and the maturation period of the wines. The implications of the variation between the different wine styles for the design of calibration models with accurate predictive abilities were investigated using glycerol calibration in wine as a model system. PCA enabled the identification and interpretation of samples that were poorly predicted by the calibration models, as well as the detection of individual samples in the sample set that had atypical spectra (i.e., outlier samples). The Soft Independent Modeling of Class Analogy (SIMCA) approach was used to establish a model for the classification of the outlier samples. A glycerol calibration for wine was developed (reducing sugar content < 30 g/L, alcohol > 8% v/v) with satisfactory predictive ability (SEP = 0.40 g/L). The RPD value (ratio of the standard deviation of the data to the standard error of prediction) was 5.6, indicating that the calibration is suitable for quantification purposes. A calibration for glycerol in special late harvest and noble late harvest wines (RS 31-147 g/L, alcohol > 11.6% v/v) with a prediction error SECV = 0.65 g/L, was also established. This study yielded an analytical strategy that combined the careful design of calibration sets with measures that facilitated the early detection and interpretation of poorly predicted samples and outlier samples in a sample set. The strategy provided a powerful means of quality control, which is necessary for the generation of accurate prediction data and therefore for the successful implementation of FT-IR in the routine analytical laboratory.


Proximate grassland and shrub-encroached sites show dramatic restructuring of soil bacterial communities.
Changes in aboveground community composition and diversity following shrub encroachment have been studied extensively. Recently, shrub encroachment was associated with differences in belowground bacterial communities relative to non-encroached grassland sites hundreds of meters away. This spatial distance between grassland and shrub sites left open the question of how soil bacterial communities associated with different vegetation types might differ within the same plot location. We examined soil bacterial communities between shrub-encroached and adjacent (one m apart) grassland soils in Chinese Inner Mongolian, using high-throughput sequencing method (Illumina, San Diego, CA, USA). Shrub-encroached sites were associated with dramatic restructuring of soil bacterial community composition and predicted metabolic function, with significant increase in bacterial alpha-diversity. Moreover, bacterial phylogenic structures showed clustering in both shrub-encroached and grassland soils, suggesting that each vegetation type was associated with a unique and defined bacterial community by niche filtering. Finally, soil organic carbon (SOC) was the primary driver varied with shifts in soil bacterial community composition. The encroachment was associated with elevated SOC, suggesting that shrub-mediated shifts in SOC might be responsible for changes in belowground bacterial community. This study demonstrated that shrub-encroached soils were associated with dramatic restructuring of bacterial communities, suggesting that belowground bacterial communities appear to be sensitive indicators of vegetation type. Our study indicates that the increased shrub-encroached intensity in Inner Mongolia will likely trigger large-scale disruptions in both aboveground plant and belowground bacterial communities across the region.


IL-17+ CD8+ T cell suppression by dimethyl fumarate associates with clinical response in multiple sclerosis.
IL-17-producing CD8+ (Tc17) cells are enriched in active lesions of patients with multiple sclerosis (MS), suggesting a role in the pathogenesis of autoimmunity. Here we show that amelioration of MS by dimethyl fumarate (DMF), a mechanistically elusive drug, associates with suppression of Tc17 cells. DMF treatment results in reduced frequency of Tc17, contrary to Th17 cells, and in a decreased ratio of the regulators RORC-to-TBX21, along with a shift towards cytotoxic T lymphocyte gene expression signature in CD8+ T cells from MS patients. Mechanistically, DMF potentiates the PI3K-AKT-FOXO1-T-BET pathway, thereby limiting IL-17 and RORγt expression as well as STAT5-signaling in a glutathione-dependent manner. This results in chromatin remodeling at the Il17 locus. Consequently, T-BET-deficiency in mice or inhibition of PI3K-AKT, STAT5 or reactive oxygen species prevents DMF-mediated Tc17 suppression. Overall, our data disclose a DMF-AKT-T-BET driven immune modulation and suggest putative therapy targets in MS and beyond.


SYNE1-related autosomal recessive cerebellar ataxia, congenital cerebellar hypoplasia, and cognitive impairment.
The spectrin repeat-containing nuclear envelope protein 1 (SYNE1) gene encodes a family of spectrin structural proteins that are associated with anchoring the plasma membrane to the actin cytoskeleton. SYNE1-related disease is most commonly reported in autosomal recessive spinocerebellar ataxia 8, which demonstrates variable age of onset with a median of 30 years of age. However pathogenic mutations in SYNE1 are also causative of arthrogryposis multiplex congenital, a severe congenital neuromuscular condition. Here in we report monozygous twins with childhood onset ataxia, cerebellar hypoplasia, dysarthria, and cognitive impairment sharing two novel heterozygous mutations in the SYNE1 gene. Our family may expand the clinical phenotype associated with SYNE1-related disease and offers possible genotype-phenotype correlations of a rare continuum of clinical disease phenotypes from neonatal to adult onset.


EpiReumaPt- the study of rheumatic and musculoskeletal diseases in Portugal: a detailed view of the methodology.
Rheumatic and musculoskeletal diseases (RMD) are prevalent and leading causes of disability and consumption of healthcare and social resources. EpiReumaPt is a national population-based survey developed by the Portuguese Society of Rheumatology that aimed to estimate the prevalence of RMDs and determine their impact on function, quality of life, mental health and use of healthcare resources. This article describes in detail the design, methodology and planned analyses of EpiReumaPt. Recruitment started in September 2011 and finished in December 2013. This study involved a three-stage approach. The first step was a face-to-face survey performed by trained interviewers at the household of 10,661 subjects who where randomly selected by a stratified multistage sampling. A highly sensitive screening questionnaire for RMDs was used. Secondly, participants who screened positive (64%) for at least one RMD as well as 20% of individuals with a negative screening were invited for assessment by a rheumatologist. In total, 3,877 subjects participated in this second phase, where they were also invited to donate a blood sample to be stored at the Biobanco-IMM. History and physical examination, followed by appropriate laboratory and imaging tests were performed. At the end of the visit, the rheumatologist established a diagnosis. Finally, a team of three experienced rheumatologists reviewed all the clinical data and defined the diagnoses according to previously validated criteria. The EpiReumaPt dataset, containing data from several questionnaires, various clinical measurements and information from laboratory and imaging tests, comprises an invaluable asset for research. The large amount of information collected from each participant and the large number of participants, with a wide age range covering and being representative of the adults from the entire country, makes EpiReumaPt the largest study of RMDs performed in Portugal.


Disenrollment of Medicare cancer patients from health maintenance organizations.
There is concern that financial incentives in health maintenance organizations (HMOs) might result in pressures to induce sicker members to disenroll. The authors compared disenrollment rates of Medicare HMO enrollees with cancer with disenrollment rates for cancer-free enrollees, using Medicare enrollment files linked to population-based tumor registry data from the Surveillance, Epidemiology, and End Results (SEER) Program. The authors identified all aged Medicare beneficiaries who enrolled in an HMO located in a SEER reporting area during 1985 to 1989. Time to disenrollment was analyzed using a proportional hazards model. Overall, cancer patients were no more likely to disenroll than others. However, persons diagnosed with cancer after enrollment were less likely to disenroll than other persons in Independent Practice Association (IPA) mode HMOs (relative risk [RR] = 0.79). Persons diagnosed with cancer after enrollment in group- and staff-model HMOs were about equally likely to disenroll as other persons (RR = 0.91). Persons diagnosed with cancer less than 18 months before enrollment were at high risk for disenrollment from both IPA and group-/staff-model HMOs (RR = 1.47 and 1.35). There was substantial variation among HMOs in overall disenrollment rates and in RRs for disenrollment by cancer patients. The low disenrollment rates of patients diagnosed after enrollment do not support the contention that features intrinsic to managed care make HMOs unattractive to the seriously ill. Monitoring of selective disenrollment could be used as a screen for possible access and quality problems.


Screening of autism based on task-free fMRI using graph theoretical approach.
Studies on autism spectrum disorder (ASD) have indicated several dysfunctions in the structure, and functional organization of the brain. However, findings have not been established as a general diagnostic tool yet. In this regard, current study proposed an automatic screening method for recognition of ASDs from healthy controls (HCs) based on their brain functional abnormalities. In this paradigm, brain functional networks of 60 adolescent and young adult males (29 ASDs and 31 HCs) were estimated from subjects' task-free fMRI data. Then, autism screening was developed based on characteristics of the functional networks using the following steps: A) local and global parameters of the brain functional network were calculated using graph theory. B) network parameters of the ASDs were statistically compared to the HCs. C) significantly altered parameters were used as input features of the screening system. D) performance of the system was verified using various classification techniques. The support vector machine showed superiority to others with an accuracy of 92%. Subsequently, reliability of the results was examined using an independent dataset including 20 ASDs and 20 HCs. Our findings suggest that local parameters of the brain functional network, despite the individual variability, can potentially be used for autism screening.