A Plasmodium falciparum FcB1-schizont-EST collection providing clues to schizont specific gene structure and polymorphism.
The Plasmodium falciparum genome (3D7 strain) published in 2002, revealed ~5,400 genes, mostly based on in silico predictions. Experimental data is therefore required for structural and functional assessments of P. falciparum genes and expression, and polymorphic data are further necessary to exploit genomic information to further qualify therapeutic target candidates. Here, we undertook a large scale analysis of a P. falciparum FcB1-schizont-EST library previously constructed by suppression subtractive hybridization (SSH) to study genes expressed during merozoite morphogenesis, with the aim of: 1) obtaining an exhaustive collection of schizont specific ESTs, 2) experimentally validating or correcting P. falciparum gene models and 3) pinpointing genes displaying protein polymorphism between the FcB1 and 3D7 strains. A total of 22,125 clones randomly picked from the SSH library were sequenced, yielding 21,805 usable ESTs that were then clustered on the P. falciparum genome. This allowed identification of 243 protein coding genes, including 121 previously annotated as hypothetical. Statistical analysis of GO terms, when available, indicated significant enrichment in genes involved in "entry into host-cells" and "actin cytoskeleton". Although most ESTs do not span full-length gene reading frames, detailed sequence comparison of FcB1-ESTs versus 3D7 genomic sequences allowed the confirmation of exon/intron boundaries in 29 genes, the detection of new boundaries in 14 genes and identification of protein polymorphism for 21 genes. In addition, a large number of non-protein coding ESTs were identified, mainly matching with the two A-type rRNA units (on chromosomes 5 and 7) and to a lower extent, two atypical rRNA loci (on chromosomes 1 and 8), TARE subtelomeric regions (several chromosomes) and the recently described telomerase RNA gene (chromosome 9). This FcB1-schizont-EST analysis confirmed the actual expression of 243 protein coding genes, allowing the correction of structural annotations for a quarter of these sequences. In addition, this analysis demonstrated the actual transcription of several remarkable non-protein coding loci: 2 atypical rRNA, TARE region and telomerase RNA gene. Together with other collections of P. falciparum ESTs, usually generated from mixed parasite stages, this collection of FcB1-schizont-ESTs provides valuable data to gain further insight into the P. falciparum gene structure, polymorphism and expression.


Exocytosis and synaptic vesicle function.
Synaptic vesicles release their vesicular contents to the extracellular space by Ca(2+)-triggered exocytosis. The Ca(2+)-triggered exocytotic process is regulated by synaptotagmin (Syt), a vesicular Ca(2+)-binding C2 domain protein. Synaptotagmin 1 (Syt1), the most studied major isoform among 16 Syt isoforms, mediates Ca(2+)-triggered synaptic vesicle exocytosis by interacting with the target membranes and SNARE/complexin complex. In synapses of the central nervous system, synaptobrevin 2, a major vesicular SNARE protein, forms a ternary SNARE complex with the plasma membrane SNARE proteins, syntaxin 1 and SNAP25. The affinities of Ca(2+)-dependent interactions between Syt1 and its targets (i.e., SNARE complexes and membranes) are well correlated with the efficacies of the corresponding exocytotic processes. Therefore, different SNARE protein isoforms and membrane lipids, which interact with Syt1 with various affinities, are capable of regulating the efficacy of Syt1-mediated exocytosis. Otoferlin, another type of vesicular C2 domain protein that binds to the membrane in a Ca(2+)-dependent manner, is also involved in the Ca(2+)-triggered synaptic vesicle exocytosis in auditory hair cells. However, the functions of otoferlin in the exocytotic process are not well understood. In addition, at least five different types of synaptic vesicle proteins such as synaptic vesicle protein 2, cysteine string protein α, rab3, synapsin, and a group of proteins containing four transmembrane regions, which includes synaptophysin, synaptogyrin, and secretory carrier membrane protein, are involved in modulating the exocytotic process by regulating the formation and trafficking of synaptic vesicles.


Sinergasilus polycolpus, a new copepod species in the ichthyoparasitofauna of Serbia and Montenegro.
The parasitic copepod Sinergasilus polycolpus was identified on the gills of bighead carp Aristichthys nobilis from 2 localities (Kladovo and Slankamen) in the Serbian part of the River Danube. This parasite is species-specific for 2 Chinese carp, the bighead carp and the silver carp Hypophthalmichthys molitrix. It was accidentally introduced into Serbia and Montenegro together with fry of these herbivorous carp intended for aquaculture and control of phytoplankton blooms. There is no record in the available literature of this parasite for European freshwaters. Our identification of S. polycolpus signals the possible spread of the infectious disease sinergasilosis in natural freshwaters and in fishponds, similar to bothriocephalosis, caused by Bothriocephalus opsariichthydis, which was introduced with the fry of various herbivorous species from the Amour River basin (USSR) into almost all countries throughout the world.


Habitual daily masseter activity of subjects with different vertical craniofacial morphology.
Little is known about the habitual daily masticatory activity of subjects with different vertical craniofacial morphology. The purpose of the present study was to compare the daily long-term muscle activity of short-face subjects with that of long-face subjects as assessed in their natural environment. Digital photographs of the facial profile were obtained from a sample of 300 subjects and the ratio between anterior total and anterior lower facial height was assessed (Vertical Facial Index: VFI). Fourteen long-face and 16 short-face subjects were selected from the opposite tails of the frequency distribution of VFI. Long-term masseter activity was monitored for 8 h d(-1) in the natural environment by means of portable one-channel electromyograph recorders over three working days. Assessments included calculation of the number of activity periods (APs) per h (N/h), their mean amplitude (Amean), and their mean duration (Dur). The maximal electromyogram activity did not differ significantly between the short-face and the long-face subjects. There was no significant difference between the two groups investigated in N/h, in Amean and Dur. The findings suggest that habitual activity of masseter muscle in the natural environment was not influenced by the vertical craniofacial morphology as assessed in the present study.


The effects of geography and spatial behavior on health care utilization among the residents of a rural region.
This analysis determines the importance of geography and spatial behavior as predisposing and enabling factors in rural health care utilization, controlling for demographic, social, cultural, and health status factors. A survey of 1,059 adults in 12 rural Appalachian North Carolina counties. This cross-sectional study used a three-stage sampling design stratified by county and ethnicity. Preliminary analysis of health services utilization compared weighted proportions of number of health care visits in the previous 12 months for regular check-up care, chronic care, and acute care across geographic, sociodemographic, cultural, and health variables. Multivariable logistic models identified independent correlates of health services utilization. Respondents answered standard survey questions. They located places in which they engaged health related and normal day-to-day activities; these data were entered into a geographic information system for analysis. Several geographic and spatial behavior factors, including having a driver's license, use of provided rides, and distance for regular care, were significantly related to health care utilization for regular check-up and chronic care in the bivariate analysis. In the multivariate model, having a driver's license and distance for regular care remained significant, as did several predisposing (age, gender, ethnicity), enabling (household income), and need (physical and mental health measures, number of conditions). Geographic measures, as predisposing and enabling factors, were related to regular check-up and chronic care, but not to acute care visits. These results show the importance of geographic and spatial behavior factors in rural health care utilization. They also indicate continuing inequity in rural health care utilization that must be addressed in public policy.


Early bilateral and massive compromise of the frontal lobes.
The frontal lobes are one of the most complex brain structures involved in both domain-general and specific functions. The goal of this work was to assess the anatomical and cognitive affectations from a unique case with massive bilateral frontal affectation. We report the case of GC, an eight-year old child with nearly complete affectation of bilateral frontal structures and spared temporal, parietal, occipital, and cerebellar regions. We performed behavioral, neuropsychological, and imaging (MRI, DTI, fMRI) evaluations. Neurological and neuropsychological examinations revealed a mixed pattern of affected (executive control/abstraction capacity) and considerably preserved (consciousness, language, memory, spatial orientation, and socio-emotional) functions. Both structural (DTI) and functional (fMRI) connectivity evidenced abnormal anterior connections of the amygdala and parietal networks. In addition, brain structural connectivity analysis revealed almost complete loss of frontal connections, with atypical temporo-posterior pathways. Similarly, functional connectivity showed an aberrant frontoparietal network and relative preservation of the posterior part of the default mode network and the visual network. We discuss this multilevel pattern of behavioral, structural, and functional connectivity results. With its unique pattern of compromised and preserved structures and functions, this exceptional case offers new constraints and challenges for neurocognitive theories.


Penicillin-Binding Protein Transpeptidase Signatures for Tracking and Predicting β-Lactam Resistance Levels in Streptococcus pneumoniae.
β-Lactam antibiotics are the drugs of choice to treat pneumococcal infections. The spread of β-lactam-resistant pneumococci is a major concern in choosing an effective therapy for patients. Systematically tracking β-lactam resistance could benefit disease surveillance. Here we developed a classification system in which a pneumococcal isolate is assigned to a "PBP type" based on sequence signatures in the transpeptidase domains (TPDs) of the three critical penicillin-binding proteins (PBPs), PBP1a, PBP2b, and PBP2x. We identified 307 unique PBP types from 2,528 invasive pneumococcal isolates, which had known MICs to six β-lactams based on broth microdilution. We found that increased β-lactam MICs strongly correlated with PBP types containing divergent TPD sequences. The PBP type explained 94 to 99% of variation in MICs both before and after accounting for genomic backgrounds defined by multilocus sequence typing, indicating that genomic backgrounds made little independent contribution to β-lactam MICs at the population level. We further developed and evaluated predictive models of MICs based on PBP type. Compared to microdilution MICs, MICs predicted by PBP type showed essential agreement (MICs agree within 1 dilution) of >98%, category agreement (interpretive results agree) of >94%, a major discrepancy (sensitive isolate predicted as resistant) rate of <3%, and a very major discrepancy (resistant isolate predicted as sensitive) rate of <2% for all six β-lactams. Thus, the PBP transpeptidase signatures are robust indicators of MICs to different β-lactam antibiotics in clinical pneumococcal isolates and serve as an accurate alternative to phenotypic susceptibility testing. The human pathogen Streptococcus pneumoniae is a leading cause of morbidity and mortality worldwide. β-Lactam antibiotics such as penicillin and ceftriaxone are the drugs of choice to treat pneumococcal infections. Some pneumococcal strains have developed β-lactam resistance through altering their penicillin-binding proteins (PBPs) and have become a major concern in choosing effective patient therapy. To systematically track and predict β-lactam resistance, we obtained the sequence signatures of PBPs from a large collection of clinical pneumococcal isolates using whole-genome sequencing data and found that these "PBP types" were predictive of resistance levels. Our findings can benefit the current era of strain surveillance when whole-genome sequencing data often lacks detailed resistance information. Using PBP positions that we found are always substituted within highly resistant strains may lead to further refinements. Sequence-based predictions are accurate and may lead to the ability to extract critical resistance information from nonculturable clinical specimens.


Impact of Providing Feed and/or Water on Performance, Physiology, and Behavior of Weaned Pigs during a 32-h Transport.
Transportation at weaning is a complex stressor made up of many factors, including withdrawal from feed and water, which can potentially negatively affect the health and welfare of pigs, especially those already experiencing weaning stress. The objective of this study was to evaluate the effect of weaning and extended transport durations (up to 32 h), with and without the provision of feed and/or water, on pig welfare. Treatment groups included: pigs neither weaned nor transported, control (CON); weaned pigs transported and provided with feed and water (T+); weaned pigs transported without feed and water (T-); weaned pigs transported with only feed (T+F); and weaned pigs transported with only water provided (TRAN+W). The effect of transport (with and without feed and/or water) on weaned pigs was assessed using behavior, performance, and physiology. After a 32-h transport period, pigs transported without water lost markedly more weight than those transported with water ( p < 0.01). Furthermore, the neutrophil to lymphocyte ratio was markedly higher in male pigs transported without water ( p < 0.05). Overall, transportation had a negative effect on pig well-being, especially when water was not provided.


Deposition Form and Bioaccessibility of Keto-carotenoids from Mamey Sapote (Pouteria sapota), Red Bell Pepper (Capsicum annuum), and Sockeye Salmon (Oncorhynchus nerka) Filet.
The ultrastructure and carotenoid-bearing structures of mamey sapote (Pouteria sapota) chromoplasts were elucidated using light and transmission electron microscopy and compared to carotenoid deposition forms in red bell pepper (Capsicum annuum) and sockeye salmon (Oncorhynchus nerka). Globular-tubular chromoplasts of sapote contained numerous lipid globules and tubules embodying unique provitamin A keto-carotenoids in a lipid-dissolved and presumably liquid-crystalline form, respectively. Bioaccessibility of sapotexanthin and cryptocapsin was compared to that of structurally related keto-carotenoids from red bell pepper and salmon. Capsanthin from bell pepper was the most bioaccessible pigment, followed by sapotexanthin and cryptocapsin esters from mamey sapote. In contrast, astaxanthin from salmon was the least bioaccessible keto-carotenoid. Thermal treatment and fat addition consistently enhanced bioaccessibility, except for astaxanthin from naturally lipid-rich salmon, which remained unaffected. Although the provitamin A keto-carotenoids from sapote were highly bioaccessible, their qualitative and quantitative in vivo bioavailability and their conversion to vitamin A remains to be confirmed.


Induced pluripotent stem cells restore function in a human cell loss model of open-angle glaucoma.
Normally, trabecular meshwork (TM) and Schlemm's canal inner wall endothelial cells within the aqueous humor outflow pathway maintain intraocular pressure within a narrow safe range. Elevation in intraocular pressure, because of the loss of homeostatic regulation by these outflow pathway cells, is the primary risk factor for vision loss due to glaucomatous optic neuropathy. A notable feature associated with glaucoma is outflow pathway cell loss. Using controlled cell loss in ex vivo perfused human outflow pathway organ culture, we developed compelling experimental evidence that this level of cell loss compromises intraocular pressure homeostatic function. This function was restored by repopulation of the model with fresh TM cells. We then differentiated induced pluripotent stem cells (iPSCs) and used them to repopulate this cell depletion model. These differentiated cells (TM-like iPSCs) became similar to TM cells in both morphology and expression patterns. When transplanted, they were able to fully restore intraocular pressure homeostatic function. This successful transplantation of TM-like iPSCs establishes the conceptual feasibility of using autologous stem cells to restore intraocular pressure regulatory function in open-angle glaucoma patients, providing a novel alternative treatment option.


The binding of Curcuma longa extract with bovine serum albumin monitored via time-resolved fluorescence.
Turmeric (Curcuma longa L.) is obtained from the rhizome of the Zingberaceae family and has a long history as an ingredient in cooking. It has been used as a dye and recently research has concentrated on its possible health benefits, specifically because of its antioxidant activity. The principal compound that is responsible for this activity is curcumin, which is present with the other curcuminoids; demethoxycurcumin and bisdemethoxycurcumin. Curcumin exhibits fluorescence and its photophysics are markedly affected by the polarity, hydrogen bonding and pH. This provides a means to examine its interaction with proteins, which is important if its potential health role is to be fully investigated. In this work, we monitor the binding kinetics using time-resolved fluorescence measurements, enabled by the use of low dead time electronics coupled with a high repetition rate excitation source and time-resolved emission spectra of the extracted curcuminoids upon interaction with bovine serum albumin. From these measurements the decay-associated spectra of the different lifetime components were obtained, which is consistent with reports of more than one binding site. Monitoring changes in these spectra with increasing temperature also allows for the denaturing of the serum albumin to be inferred.


Incidence of surgical site infection after spine surgery: what is the impact of the definition of infection?
Orthopaedic surgical site infections (SSIs) can delay recovery, add impairments, and decrease quality of life, particularly in patients undergoing spine surgery, in whom SSIs may also be more common. Efforts to prevent and treat SSIs of the spine rely on the identification and registration of these adverse events in large databases. The effective use of these databases to answer clinical questions depends on how the conditions in question, such as infection, are defined in the databases queried, but the degree to which different definitions of infection might cause different risk factors to be identified by those databases has not been evaluated. The purpose of this study was to determine whether different definitions of SSI identify different risk factors for SSI. Specifically, we compared the International Classification of Diseases, 9th Revision (ICD-9) coding, Centers for Disease Control and Prevention (CDC) criteria for deep infection, and incision and débridement for infection to determine if each is associated with distinct risk factors for SSI. In this single-center retrospective study, a sample of 5761 adult patients who had an orthopaedic spine surgery between January 2003 and August 2013 were identified from our institutional database. The mean age of the patients was 56 years (± 16 SD), and slightly more than half were men. We applied three different definitions of infection: ICD-9 code for SSI, the CDC criteria for deep infection, and incision and débridement for infection. Three hundred sixty-one (6%) of the 5761 surgeries received an ICD-9 code for SSI within 90 days of surgery. After review of the medical records of these 361 patients, 216 (4%) met the CDC criteria for deep SSI, and 189 (3%) were taken to the operating room for irrigation and débridement within 180 days of the day of surgery. We found the Charlson Comorbidity Index, the duration of the operation, obesity, and posterior surgical approach were independently associated with a higher risk of infection for each of the three definitions of SSI. The influence of malnutrition, smoking, specific procedures, and specific surgeons varied by definition of infection. These elements accounted for approximately 6% of the variability in the risk of developing an infection. The frequency of SSI after spine surgery varied according to the definition of an infection, but the most important risk factors did not. We conclude that large database studies may be better suited for identifying risk factors than for determining absolute numbers of infections. Level III, prognostic study. See Guidelines for Authors for a complete description of levels of evidence.


Mesenchymal stromal cells shape the MDS microenvironment by inducing suppressive monocytes that dampen NK cell function.
Altered BM hematopoiesis and immune suppression are hallmarks of myelodysplastic syndrome (MDS). While the BM microenvironment influences malignant hematopoiesis, the mechanism leading to MDS-associated immune suppression is unknown. We tested whether mesenchymal stromal cells (MSCs) contribute to this process. Here, we developed a model to study cultured MSCs from patients with MDS (MDS-MSCs) compared with those from aged-matched normal controls for regulation of immune function. MDS-MSCs and healthy donor MSCs (HD-MSCs) exhibited a similar in vitro phenotype, and neither had a direct effect on NK cell function. However, when MDS- and HD-MSCs were cultured with monocytes, only the MDS-MSCs acquired phenotypic and metabolic properties of myeloid-derived suppressor cells (MDSCs), with resulting suppression of NK cell function, along with T cell proliferation. A MSC transcriptome was observed in MDS-MSCs compared with HD-MSCs, including increased expression of the ROS regulator, ENC1. High ENC1 expression in MDS-MSCs induced suppressive monocytes with increased INHBA, a gene that encodes for a member of the TGF-β superfamily of proteins. These monocytes also had reduced expression of the TGF-β transcriptional repressor MAB21L2, further adding to their immune-suppressive function. Silencing ENC1 or inhibiting ROS production in MDS-MSCs abrogated the suppressive function of MDS-MSC-conditioned monocytes. In addition, silencing MAB21L2 in healthy MSC-conditioned monocytes mimicked the MDS-MSC-suppressive transformation of monocytes. Our data demonstrate that MDS-MSCs are responsible for inducing an immune-suppressive microenvironment in MDS through an indirect mechanism involving monocytes.


Risk of type 2 diabetes and obesity is differentially associated with variation in FTO in whites and African-Americans in the ARIC study.
Single nucleotide polymorphisms (SNPs) in the fat mass and obesity associated (FTO) gene are associated with body mass index (BMI) in populations of European descent. The FTO rs9939609 variant, first detected in a genome-wide association study of diabetes, conferred an increased disease risk that was abolished after adjustment for BMI, suggesting that the association may be due to variation in adiposity. The relationship between diabetes, four previously identified FTO polymorphisms that span a 19.6-kb genomic region, and obesity was therefore evaluated in the biracial population-based Atherosclerosis Risk in Communities Study with the goal of further refining the association by comparing results between the two ethnic groups. The prevalence of diabetes and obesity (BMI > or = 30 kg/m(2)) was established at baseline, and diabetes was determined by either self-report, a fasting glucose level > or = 126 mg/dL, or non-fasting glucose > or = 200 mg/dL. There were 1,004 diabetes cases and 10,038 non-cases in whites, and 670 cases and 2,780 non-cases in African-Americans. Differences in mean BMI were assessed by a general linear model, and multivariable logistic regression was used to predict the risk of diabetes and obesity. For white participants, the FTO rs9939609 A allele was associated with an increased risk of diabetes (odds ratio (OR) = 1.19, p<0.001) and obesity (OR = 1.22, p<0.001) under an additive genetic model that was similar for all of the SNPs analyzed. In African-Americans, only the rs1421085 C allele was a determinant of obesity risk (OR = 1.17, p = 0.05), but was found to be protective against diabetes (OR = 0.79, p = 0.03). Adjustment for BMI did not eliminate any of the observed associations with diabetes. Significant statistical interaction between race and the FTO variants suggests that the effect on diabetes susceptibility may be context dependent.


Effect of prolonged coldness on survival and fertility of Drosophila melanogaster.
The laboratory fruit fly, Drosophila melanogaster, is used widely in biological research, but the requirement to maintain stocks with a roughly biweekly generation time imposes substantial burdens of labor, potential cross-contamination and mutation accumulation. The purpose of this study was to assess the impact of prolonged cold stress or milder cooling on survivorship and fertility. The hypothesis was that cold storage would result in postponement of reproduction and a longer generation time. Flies of several genotypes were maintained continuously at 4-11 °C; recovery rates and subsequent yields of adult progeny were recorded. Adults and pupae of a relatively long-lived y w lineage were more resistant to severe cold stress than embryos and larvae. Adults exhibited minimal mortality up to at least 5 d at 4 °C, 20 d at 8 °C and 12 weeks at 11 °C. Reproduction did not occur at these temperatures, but progeny were obtained after recovery at 25 °C. At all temperatures, chilling caused a rapid, severe and progressive decrease in fertility during the first 2 d of recovery. The impact on fertility during the subsequent 2-4 d was much milder and it occurred only after prolonged incubation at low temperatures. The total reproductive output during the first 6 d of recovery was sufficient to replace the parental population after 12 weeks at 11 °C. Food spoilage had an unexpectedly low impact on survivorship and fertility, and the reproductive output of F1 progeny was not affected by storing parental flies at 11 °C for 8-10 weeks. In the case of w1118 flies, replacement of the parents within 6 d of recovery was possible for up to 60 d at 11 °C. Among less fertile genotypes, replacement of the parents was possible within 18 d after 4-10 weeks at 11 °C. These results show that the 2-week maintenance interval of stocks of D. melanogaster can be extended 3-7 fold, at least for 1 generation, by storing adult flies at 11 °C.


Evidence that Ser87 of BimEL is phosphorylated by Akt and regulates BimEL apoptotic function.
Bim, the Bcl-2 interacting mediator of cell death, is a member of the BH3-only family of pro-apoptotic proteins. Recent studies have demonstrated that the apoptotic activity of Bim can be regulated through a post-translational mechanism whereby ERK phosphorylation serves as a signal for Bim ubiquitination and proteasomal degradation. In this report, we investigated the signaling pathways leading to Bim phosphorylation in Ba/F3 cells, an interleukin-3 (IL-3)-dependent B-cell line. IL-3 stimulation induced phosphorylation of Bim(EL), one of the predominant isoforms of Bim expressed in cells, at multiple sites, as evidenced by the formation of at least three to four bands by Western blotting that were sensitive to phosphatase digestion. The appearance of multiple, phosphorylated species of Bim(EL) correlated with Akt, and not ERK, activation. The PI3K inhibitor, LY294002, blocked IL-3-stimulated Akt activity and partially blocked Bim(EL) phosphorylation. In vitro kinase assays showed that recombinant Akt could directly phosphorylate a GST-Bim(EL) fusion protein and identified the Akt phosphorylation site in the Bim(EL) domain as Ser(87). Further, we demonstrated that cytokine stimulation promotes Bim(EL) binding to 14-3-3 proteins. Finally, we show that mutation of Ser(87) dramatically increases the apoptotic potency of Bim(EL). We propose that Ser(87) of Bim(EL) is an important regulatory site that is targeted by Akt to attenuate the pro-apoptotic function of Bim(EL), thereby promoting cell survival.


Comparable efficacy of standardized Ayurveda formulation and hydroxychloroquine sulfate (HCQS) in the treatment of rheumatoid arthritis (RA): a randomized investigator-blind controlled study.
Hydroxychloroquine sulfate (HCQS) is a popular disease-modifying antirheumatic drug (DMARD) despite modest efficacy and toxicity. Ayurveda (ancient India medicinal system) physicians treat rheumatoid arthritis (RA) with allegedly safer herbal formulations. We report a head-to-head comparison in an exploratory drug trial. The objective is to compare standardized Ayurvedic formulations and HCQS in the treatment of RA. One hundred twenty-one patients with active moderately severe RA (ACR 1988 classified) were randomized into a 24-week investigator-blind, parallel efficacy, three-arm (two Ayurvedic and HCQS) multicenter drug trial study; polyherb (Tinospora cordifolia and Zingiber officinale based) and monoherb (Semecarpus anacardium). Study measures included joint counts (pain/tenderness and swelling), pain visual analogue scale, global disease assessments, and health assessment questionnaire. Oral meloxicam (fixed-dosage schedule) was prescribed to all patients during the initial 16 weeks. Patients on prednisolone could continue a fixed stable dose (<7.5 mg daily). Rescue oral use of paracetamol was permitted and monitored. All groups matched well at baseline. An intent-to-treat analysis (ANOVA, significance P < 0.05) did not show significant differences by treatment groups. In the polyherb, monoherb, and HCQS arms, 44%, 36%, and 51%, respectively, showed ACR 20 index improvement. Several efficacy measures improved significantly in the HCQS and polyherb groups with no difference between the groups (corrected P). However, the latter was individually superior to monoherb. Only mild adverse events (gut and skin, and none withdrew) were reported with no differences between the groups. Forty-two patients dropped out. This preliminary drug trial controlled for HCQS demonstrated a standardized Ayurvedic polyherb drug to be effective and safe in controlling active RA. A better-designed study with a longer evaluation period is recommended.


Antibiotic Resistance Is Associated with Integrative and Conjugative Elements and Genomic Islands in Naturally Circulating Streptococcus pneumoniae Isolates from Adults in Liverpool, UK.
Pneumonia is the sixth largest cause of death in the UK. It is usually caused by Streptococcus pneumoniae, which healthy individuals can carry in their nose without symptoms of disease. Antimicrobial resistance further increases mortality and morbidity associated with pneumococcal infection, although few studies have analysed resistance in naturally circulating pneumococcal isolates in adult populations. Here, we report on the resistome and associated mobile genetic elements within circulating pneumococcus isolated from adult volunteers enrolled in the experimental human pneumococcal colonisation (EHPC) research program at the Liverpool School of Tropical Medicine, UK. Pneumococcal isolates collected from 30 healthy asymptomatic adults who had volunteered to take part in clinical research were screened for antibiotic susceptibility to erythromycin and tetracycline, and whole-genome sequenced. The genetic context of resistance to one or both antibiotics in four isolates was characterised bioinformatically, and any association of the resistance genes with mobile genetic elements was determined. Tetracycline and macrolide resistance genes [tet(M), erm(B), mef(A), msr(D)] were detected on known Tn916-like integrative and conjugative elements, namely Tn6002 and Tn2010, and tet(32) was found for the first time in S. pneumoniae located on a novel 50 kb genomic island. The widespread use of pneumococcal conjugate vaccines impacts on serotype prevalence and transmission within the community. It is therefore important to continue to monitor antimicrobial resistance (AMR) genes present in both vaccine types and non-vaccine types in response to contemporary antimicrobial therapies and characterise the genetic context of acquired resistance genes to continually optimise antibiotic therapies.


LiST as a catalyst in program planning: experiences from Burkina Faso, Ghana and Malawi.
African countries are working to achieve rapid reductions in maternal and child mortality and meet their targets for the Millennium Development Goals (MDGs). Partners in the Catalytic Initiative to Save One Million Lives (CI) are assisting them by providing funding and technical assistance to increase and accelerate coverage for proven interventions. Here we describe how the Lives Saved Tool (LiST) was used as part of an early assessment of the expected impact of CI plans in Malawi, Burkina Faso and Ghana. LiST builds on country-specific demographic and cause-of-death profiles, and models the effect of changes in coverage for proven interventions on future levels of mortality among children less than 5 years of age. We worked with representatives of Ministries of Health and their development partners to apply LiST to assess the potential impact of CI plans and coverage targets, generating a short list of the highest-priority interventions for additional scale-up to achieve rapid reductions in under-5 mortality. The results show that in each country, achieving national coverage targets for just four or five high-impact interventions could reduce under-5 mortality by at least 20% by 2011, relative to 2006 levels. Even greater gains could be obtained in Burkina Faso and Ghana by scaling up these high-impact interventions to 80%. Discussion LiST can contribute to the development of stronger programmes by identifying the highest-impact interventions in a given epidemiological setting. The quality of LiST estimates is dependent on the available data on coverage levels and causes of death, and assumes that the target levels of coverage are feasible in a given context while maintaining service quality. Further experience is needed in the feasibility and usefulness of LiST as part of the program planning process at district and subdistrict levels.


Ecology of Indigenous Soil Rhizobia: Response of Bradyrhizobium japonicum to Readily Available Substrates.
Populations of indigenous Bradyrhizobium japonicum serocluster 123 and serogroups 110 and 138 were studied after various sugars were added to their soil habitat. Loam soil with approximately 10 cells of each group per g of soil were amended every 3 days with 0.1% glucose, sucrose, arabinose, xylose, or galactose. Enumerations of the populations were made every 12 days by immunofluorescence assay. Each B. japonicum population in the sugar-treated soils increased by about 1 log during the first 12 days, to a maximum of about 10 cells by day 36 or 48, irrespective of the sugar added. Maximum growth rates were similar for each group and occurred during the 12-day incubation period. The most rapid growth was in response to arabinose, with a mean generation time of about 3.0 days. Other mean doubling times were 4.0 days with glucose and galactose treatments, 4.5 days with xylose treatment, and 5.4 days with sucrose amendment. These data provide the first direct evidence that indigenous soil rhizobia can compete successfully with other soil bacteria for readily available substrates in soil in the absence of host legume roots or other rhizospheres. The growth rates in soil of the specific B. japonicum populations studied were nearly the same with a given sugar treatment but varied considerably with different sugars. The mean generation times of 3 to 5 days are among the first reported growth rates for heterotrophic bacteria in natural soil.


The 5' flanking region of human epsilon-globin gene.
The structural analysis of the 2.0 kb region upstream from the epsilon-globin gene has been carried out. A genomic DNA map around the gene was worked out in some detail to ensure that the cloned DNA was representative of the actual chromosomal arrangement. Furthermore, a new technique was developed to precisely map a reiterated DNA sequence present 1.5 kb to the 5' side of the gene. The complete nucleotide sequence of the 2.0 kb 5' flanking region was then determined and overlapped with the gene. The sequence included the reiterated DNA sequence which is homologous to the so-called AluI family of repeats. Unusual stretches of sequence 50 nucleotides long, where A + T represent about 90% of the bases, are present at both the 5' and 3' sides of the repeat.


Two distinct cellular mechanisms of osteoclast formation and bone resorption in periprosthetic osteolysis.
TNFalpha and IL-1alpha are proinflammatory cytokines that are abundant in periprosthetic tissues. These cytokines stimulate bone resorption and have recently been shown to directly induce osteoclast formation in mouse marrow cultures. We examined whether TNFalpha and IL-1alpha can directly induce osteoclast formation from human arthroplasty-derived (CD14(+)) macrophages by a mechanism independent of RANKL-induced osteoclastogenesis. TNFalpha and M-CSF (+/-IL-1alpha) were added to cultures of magnetically sorted (CD14(+)) and unsorted (CD14(+)/CD14(-)) cells isolated from the pseudomembrane of loosened hip arthroplasties. Osteoprotegerin (OPG), RANK:Fc and antibodies to TNF receptors (p55 and p75) were added to these cultures to distinguish the pathway of osteoclastogenesis. Osteoclast differentiation was assessed by expression of tartrate-resistant acid phosphatase (TRAP), vitronectin receptor (VNR) and lacunar resorption. The addition of TNFalpha (+/-IL-1alpha) resulted in differentiation of CD14(+) macrophages into TRAP(+) and VNR(+) multinucleated cells capable of extensive lacunar resorption. Both OPG and RANK:Fc (which inhibit RANKL-induced osteoclastogenesis) did not block osteoclastogenesis. The addition of antibodies directed against the p55 receptor subunit of TNF resulted in significant inhibition of osteoclast formation and lacunar resorption. Our results indicate that, in the presence of M-CSF, TNFalpha is sufficient for inducing human osteoclast differentiation from arthroplasty macrophages and that TNFalpha acts synergistically with IL-1alpha to stimulate lacunar resorption. This process is distinct from the RANK/RANKL signalling pathway and is likely to operate in periprosthetic tissues when there is heavy wear particle deposition and cytokine production.


Algorithmic mechanisms for reliable crowdsourcing computation under collusion.
We consider a computing system where a master processor assigns a task for execution to worker processors that may collude. We model the workers' decision of whether to comply (compute the task) or not (return a bogus result to save the computation cost) as a game among workers. That is, we assume that workers are rational in a game-theoretic sense. We identify analytically the parameter conditions for a unique Nash Equilibrium where the master obtains the correct result. We also evaluate experimentally mixed equilibria aiming to attain better reliability-profit trade-offs. For a wide range of parameter values that may be used in practice, our simulations show that, in fact, both master and workers are better off using a pure equilibrium where no worker cheats, even under collusion, and even for colluding behaviors that involve deviating from the game.


Lipid Extraction by α-Synuclein Generates Semi-Transmembrane Defects and Lipoprotein Nanoparticles.
Modulations of synaptic membranes play an essential role in the physiological and pathological functions of the presynaptic protein α-synuclein (αSyn). Here we used solution atomic force microscopy (AFM) and electron paramagnetic resonance (EPR) spectroscopy to investigate membrane modulations caused by αSyn. We used several lipid bilayers to explore how different lipid species may regulate αSyn-membrane interactions. We found that at a protein-to-lipid ratio of ∼1/9, αSyn perturbed lipid bilayers by generating semi-transmembrane defects that only span one leaflet. In addition, αSyn coaggregates with lipid molecules to produce ∼10 nm-sized lipoprotein nanoparticles. The obtained AFM data are consistent with the apolipoprotein characteristic of αSyn. The role of anionic lipids was elucidated by comparing results from zwitterionic and anionic lipid bilayers. Specifically, our AFM measurements showed that anionic bilayers had a larger tendency of forming bilayer defects; similarly, our EPR measurements revealed that anionic bilayers exhibited more substantial changes in lipid chain mobility and bilayer polarity. We also studied the effect of cholesterol. We found that cholesterol increased the capability of αSyn in inducing bilayer defects and altering lipid chain mobility and bilayer polarity. These data can be explained by an increase in the lipid headgroup-headgroup spacing and/or specific cholesterol-αSyn interactions. Interestingly, we found an inhibitory effect of the cone-shaped phosphatidylethanolamine lipids on αSyn-induced bilayer remodeling. We explained our data by considering interlipid hydrogen-bonding that can stabilize bilayer organization and suppress lipid extraction. Our results of lipid-dependent membrane modulations are likely relevant to αSyn functioning.


Natriuretic peptide-augmented induction of nitric oxide synthase through cyclic guanosine 3',5'-monophosphate elevation in vascular smooth muscle cells.
To elucidate the role of natriuretic peptides in vascular remodeling, the effects of atrial natriuretic peptide, brain natriuretic peptide, and C-type natriuretic peptide (CNP) on the induction of inducible nitric oxide (NO) synthase (iNOS) in rat aortic smooth muscle cells were examined. Although none of the peptides when applied alone induced the production of nitrite, a stable end product of NO, each peptide dramatically enhanced nitrite production induced by a cytokine combination of interleukin-1 alpha and tumor necrosis factor-alpha. Each natriuretic peptide stimulated intracellular cGMP accumulation in a dose-dependent manner. Time-dependent nitrite production by the cytokines was increased by CNP cotreatment and inhibited by NG-methyl-L-arginine, indicating involvement of the L-arginine-NO pathway. Northern blot analysis showed that the augmented nitrite production was accompanied by an increase in iNOS messenger RNA. A cGMP analog, 8-bromo-cGMP, completely mimicked all of the effects of CNP described above. A cGMP-dependent protein kinase inhibitor, KT5823, paradoxically increased nitrite production and iNOS messenger RNA levels induced by the combination of 8-bromo-cGMP and both cytokines or by the two cytokines only. These data demonstrate the stimulatory effect of cGMP on cytokine-induced iNOS and imply that natriuretic peptides may play a regulatory role in vascular remodeling via the production of large amounts of NO.


Dynamics of the cellular metabolome during human cytomegalovirus infection.
Viral replication requires energy and macromolecular precursors derived from the metabolic network of the host cell. Despite this reliance, the effect of viral infection on host cell metabolic composition remains poorly understood. Here we applied liquid chromatography-tandem mass spectrometry to measure the levels of 63 different intracellular metabolites at multiple times after human cytomegalovirus (HCMV) infection of human fibroblasts. Parallel microarray analysis provided complementary data on transcriptional regulation of metabolic pathways. As the infection progressed, the levels of metabolites involved in glycolysis, the citric acid cycle, and pyrimidine nucleotide biosynthesis markedly increased. HCMV-induced transcriptional upregulation of specific glycolytic and citric acid cycle enzymes mirrored the increases in metabolite levels. The peak levels of numerous metabolites during infection far exceeded those observed during normal fibroblast growth or quiescence, demonstrating that HCMV markedly disrupts cellular metabolic homeostasis and institutes its own specific metabolic program.


Enhanced neural activation with blueberry supplementation in mild cognitive impairment.
Preclinical studies have shown that blueberry supplementation can improve cognitive performance and neural function in aged animals and have identified associations between anthocyanins and such benefits. Preliminary human trials also suggest cognitive improvement in older adults, although direct evidence of enhancement of brain function has not been demonstrated. In this study, we investigated the effect of blueberry supplementation on regional brain activation in older adults at risk for dementia. In a randomized, double-blind, placebo-controlled trial we performed pre- and post-intervention functional magnetic resonance imaging during a working memory (WM) task to assess the effect of blueberry supplementation on blood oxygen level-dependent (BOLD) signal in older adults with mild cognitive impairment, a risk condition for dementia. Following daily supplementation for 16 weeks, blueberry-treated participants exhibited increased BOLD activation in the left pre-central gyrus, left middle frontal gyrus, and left inferior parietal lobe during WM load conditions (corrected P < 0.01). There was no clear indication of WM enhancement associated with blueberry supplementation. Diet records indicated no between-group difference in anthocyanin consumption external to the intervention. These data demonstrate, for the first time, enhanced neural response during WM challenge in blueberry-treated older adults with cognitive decline and are consistent with prior trials showing neurocognitive benefit with blueberry supplementation in this at-risk population.


On the Spike Train Variability Characterized by Variance-to-Mean Power Relationship.
We propose a statistical method for modeling the non-Poisson variability of spike trains observed in a wide range of brain regions. Central to our approach is the assumption that the variance and the mean of interspike intervals are related by a power function characterized by two parameters: the scale factor and exponent. It is shown that this single assumption allows the variability of spike trains to have an arbitrary scale and various dependencies on the firing rate in the spike count statistics, as well as in the interval statistics, depending on the two parameters of the power function. We also propose a statistical model for spike trains that exhibits the variance-to-mean power relationship. Based on this, a maximum likelihood method is developed for inferring the parameters from rate-modulated spike trains. The proposed method is illustrated on simulated and experimental spike trains.


Adapted CBT to Stabilize Sleep on Psychiatric Wards: a Transdiagnostic Treatment Approach.
Almost all patients admitted at acute crisis to a psychiatric ward experience clinically significant symptoms of insomnia. Ward environments pose challenges to both sleep and the delivery of therapy. Despite this, there is no description of how to adapt cognitive behavioural therapy (CBT) for insomnia to overcome these challenges. (i) To describe the key insomnia presentations observed in the Oxford Ward Sleep Solution (OWLS) trial and (ii) outline key adaptations aimed to increase accessibility and hence effectiveness of CBT for insomnia for a ward setting. Trial therapists collaboratively agreed the key insomnia presentations and therapy adaptations based on their individual reflective logs used during the trial. Three key insomnia presentations are outlined. These are used to illustrate the application of 10 CBT for insomnia therapy adaptations. These include use of sleep monitoring watches to engage patients in treatment, stabilizing circadian rhythms, reducing the impact of night-time observations and managing discharge as a sleep challenge. Whilst inpatient wards bring challenges for sleep and therapy delivery, creative adaptations can increase the accessibility of evidence based CBT for insomnia techniques. This therapy has proven popular with patients.


Male genitalia of neotropical Charaxinae: a comparative analysis of character variation.
Charaxinae (Lepidoptera: Nymphalidae) has a worldwide distribution, although it occurs mainly between the tropics. Most species occur in the Neotropics, where three tribes, Preponini, Anaeomorphini, and Anaenini, can be found. Collectively, these three tribes encompass 109 species. Because of its relevance to systematics and taxonomy, the male genitalia of Lepidoptera have been extensively studied. The male genitalia are composed of the last two abdominal segments and their modifications for mating, known as claspers of the bodies. In order to improve upon the systematic classification of the subfamily, 31 species of 13 genera of Neotropical Charaxinae were analyzed. All characters relevant to species and generic taxonomy were analyzed. Most structures showed morphological variations among tribes, genera, and species. These variations demonstrated to be important to Preponini, because the structural patterns of the genitalia allow the separation in two groups, Prepona Boisduval and Archaeoprepona Fruhstorfer, and are in accord with the recent taxonomic classification proposed by Ortiz-Acevado and Willmott ( 2013 ), wherein Agrias Doubleday is synonymized in Prepona and Noreppa Rydon within Archaeoprepona. In the same way, Anaeomorpha splendida Rothschild showed considerable differences from Preponini's genera, the tribe in which it was included, confirming the revalidation of the tribe Anaeomorphini ( Ortiz-Acevado and Willmott 2013 ). Substantial variation was found in the genital structures of Anaeini, making it difficult to establish structural patterns for this group. Such structural variation, however, may be very efficient to diagnose species, such as some species of Memphis Hübner and Fountainea Rydon, which can be easily identified through the presence and location of spines on the valva.


Predictors of students' success in community college nursing programs.
Many conceptual models have been applied in the investigation of college retention of nursing students. We tested a model that specifies four general constructs as predictors of student success in nursing education-dispositional factors, career value factors, situational factors, and institutional factors. The purpose of this article is to describe predictors of nursing students' success, specifically: (a) What factors predict success of nursing students graduating from community colleges in California? and (b) What factors predict success of nursing students graduating on time from community colleges in California? The study design was correlational and descriptive in nature, with a convenience sample of six intervention colleges and six matched-pair control colleges. Results of the logistic regressions indicated on-time and any-time graduations were predicted by higher grade point averages in prenursing and science. Higher prenursing grades are positive predictors of graduation; improvements in performance prior to commencing nursing education should improve student success.


Sequence of electrode implantation and outcome of deep brain stimulation for Parkinson's disease.
The effect of the variability of electrode placement on outcomes after bilateral deep brain stimulation of subthalamic nucleus has not been sufficiently studied, especially with respect to the sequence of hemisphere implantation. We retrospectively analysed the clinical and radiographic data of all the consecutive patients with Parkinson's disease who underwent surgery at our centre and completed at least 1 year follow-up. The dispersion in electrode location was calculated by the square of deviation from population mean, and the direction of deviation was analysed by comparing the intended and final implantation coordinates. Linear regression analysis was performed to analyse the predictors of postoperative improvement of the motor condition, also controlling for the sequence of implanted hemisphere. 76 patients (mean age 58±7.2 years) were studied. Compared with the first side, the second side electrode tip had significantly higher dispersion as an overall effect (5.6±21.6 vs 2.2±4.9 mm(2), p=0.04), or along the X-axis (4.1±15.6 vs 1.4±2.4 mm(2), p=0.03) and Z-axis (4.9±11.5 vs 2.9±3.6 mm(2), p=0.02); the second side stimulation was also associated with a lower threshold for side effects (contact 0, p<0.001 and contact 3, p=0.004). In the linear regression analysis, the significant predictors of outcome were baseline activities of daily living (p=0.010) and dispersion of electrode on the second side (p=0.005). We observed a higher dispersion for the electrode on the second implanted side, which also resulted to be a significant predictor of motor outcome at 1 year.


[Francesco Matiassi's 1908 design of the main city hospital of Rijeka].
In 1908, the City of Rijeka embarked on what would have been the biggest project ever. It planned to build a new city hospital that was to relieve the burden from the Saint Spirit Hospital, which could no longer meet local healthcare needs. The hospital was designed by one of Rijeka's finest architects, Francesco Matiassi and was to have 600 beds and cover an area of 40,000 sq m. In his design, Mattiassi implemented the programme and suggestions of the hospital head, Dr Antonio Grossich and the City technical department, headed by Luigi Luppis. The hospital complex was to be a combination of two types of buildings, common in hospital architecture of the time: the main building with a very complex floor plan, and four separate pavilions: mental illnesses, quarantine, post mortem, and services. The first three floors of the main building were intended for the departments of general medicine, surgery, gynaecology, maternity, infectious diseases, ophthalmology, and dermatology, and the remaining floors for patient reception, hospital supplies and utilities, administration, and staff. Glass-walled operating theatres, terraces for patients and facilities such as a large cloakroom bear witness to the designer's deep understanding of civil culture and architecture, which set high standards even in today's terms. Even though the project was too ambitious and even utopian for a city like Rijeka at the beginning of the 20th century, we cannot but regret that it has not seen the light of day at least in part.


Sodium iron EDTA [NaFe(III)EDTA] as a food fortificant: erythrocyte incorporation of iron and apparent absorption of zinc, copper, calcium, and magnesium from a complementary food based on wheat and soy in healthy infants.
Phytic acid is a strong inhibitor of iron absorption from fortified foods. In adults, this inhibitory effect can be overcome by adding ascorbic acid with the iron fortificant or by using a "protected" iron compound such as NaFeEDTA. In addition, the use of NaFeEDTA as an iron fortificant has been reported to increase zinc absorption in adult women. No information is available on iron bioavailability from NaFeEDTA or the influence of NaFeEDTA on minerals and trace elements in infants. We aimed to compare iron bioavailability from a complementary food based on wheat and soy fortified with either NaFeEDTA or ferrous sulfate plus ascorbic acid. The apparent absorption of zinc, copper, calcium, and magnesium was evaluated in parallel. Stable-isotope techniques were used in a crossover design to evaluate erythrocyte incorporation of iron 14 d after administration of labeled test meals and the apparent absorption of zinc, copper, calcium, and magnesium on the basis of fecal monitoring in 11 infants. Geometric mean erythrocyte incorporation of iron was 3.7% (NaFeEDTA) and 4.9% (ferrous sulfate plus ascorbic acid) (P = 0.08). No significant differences in the apparent absorption of zinc, copper, calcium, or magnesium were observed between test meals (n = 10). Iron bioavailability from a high-phytate, cereal-based complementary food fortified with either NaFeEDTA or ferrous sulfate plus ascorbic acid was not significantly different. NaFeEDTA did not influence the apparent absorption of zinc, copper, calcium, or magnesium. NaFeEDTA does not provide any nutritional benefit compared with the combination of a highly bioavailable iron compound and ascorbic acid.


Growth hormone and dexamethasone stimulate lipolysis and activate adenylyl cyclase in rat adipocytes by selectively shifting Gi alpha2 to lower density membrane fractions.
GH, in the presence of glucocorticoid, produces a delayed increase in lipolysis in rat adipose tissue, but the biochemical mechanisms that account for this action have not been established. Other lipolytic agents rapidly activate adenylyl cyclase (AC) and the resulting production of cAMP initiates a chain of reactions that culminates in the activation of hormone-sensitive lipase. We compared responses of segments of rat epididymal fat or isolated adipocytes to 30 ng/ml GH and 0.1 microg/ml dexamethasone (Dex) with 0.1 ng/ml isoproterenol (ISO), which evoked a similar increase in lipolysis. All measurements were made during the fourth hour after the addition of GH+Dex or immediately after the addition of ISO to cells or tissues that had been preincubated for 3 h without hormone. Although no significant increases in cAMP were discernible in homogenates of GH+Dex-treated tissues, Rp-cAMPS (Rp-adenosine 3'5'-phosphothioate), a competitive inhibitor of cAMP, was equally effective in decreasing lipolysis induced by GH+Dex or ISO. The proportion of PKA that was present in the active form was determined by measuring the incorporation of 32P from [gamma-32P]ATP into kemptide in the absence and presence of saturating amounts of cAMP. GH+Dex and ISO produced similar increases in protein kinase A activity in tissue extracts. Treatment with GH+Dex did not change the total forskolin-stimulated AC present in either a crude membrane pellet sedimented at 16K x g or a less dense membrane pellet sedimented at 100K x g, but doubled the AC activity in the 16K pellet when assayed in the absence of forskolin. To evaluate possible effects on G proteins, pellets obtained from centrifugation of adipocyte homogenates at 16K x g and 100K x g were solubilized and subjected to PAGE and Western analysis. GH+Dex decreased Gi alpha2 by 44% (P < 0.02) in the 16K pellets and increased it by 52% (P < 0.01) in the 100K pellets. Gs alpha in the 16K pellet was unaffected by GH+Dex and was decreased (P < 0.05) in the 100K pellet. Sucrose density fractionation of the 16K pellets revealed a similar GH+Dex-dependent shift of Gi alpha2 to less dense fractions as determined by both Western analysis and [32P]NAD ribosylation catalyzed by pertussis toxin. No such changes were seen in the distribution of Gs alpha or 5'-nucleotidase. Colchicine (100 microM) blocked the GH+Dex-dependent shift of Gi alpha2 from the 16K to the 100K pellet and blocked the lipolytic effects of GH+Dex, but not those of ISO. We conclude that by modifying the relationship between AC and Gi alpha2, GH+Dex relieves some inhibition of cAMP production and consequently increases lipolysis.


A method to estimate biomechanics and mechanical properties of optic nerve head tissues from parameters measurable using optical coherence tomography.
Optic nerve head (ONH) tissue properties and biomechanics remain mostly unmeasurable in the experiment. We hypothesized that these can be estimated numerically from ocular parameters measurable in vivo with optical coherence tomography (OCT). Using parametric models representing human ONHs we simulated acute intraocular pressure (IOP) increases (10 mmHg). Statistical models were fit to predict, from OCT-measurable parameters, 15 outputs, including ONH tissue properties, stresses, and deformations. The calculations were repeated adding parameters that have recently been proposed as potentially measurable with OCT. We evaluated the sensitivity of the predictions to variations in the experimental parameters. Excellent fits were obtained to predict all outputs from the experimental parameters, with cross-validated R2s between 0.957 and 0.998. Incorporating the potentially measurable parameters improved fits significantly. Predictions of tissue stiffness were accurate to within 0.66 MPa for the sclera and 0.24 MPa for the lamina cribrosa. Predictions of strains and stresses were accurate to within 0.62% and 4.9 kPa, respectively. Estimates of ONH biomechanics and tissue properties can be obtained quickly from OCT measurements using an applet that we make freely available. These estimates may improve understanding of the eye sensitivity to IOP and assessment of patient risk for development or progression of glaucoma.


Recent trends in neuroendovascular therapy in Japan: analysis of a nationwide survey--Japanese Registry of Neuroendovascular Therapy (JR-NET) 1 and 2.
The present study retrospectively analyzed the database of the Japanese Registry of Neuroendovascular Therapy 1 and 2 (JR-NET1&2) to determine annual trends, including adverse events and clinical outcomes at 30 days after undergoing neuroendovascular therapy. JR-NET1&2 are surveys that targeted all patients in Japan who underwent neuroendovascular therapy delivered by physicians certified by the Japanese Society of Neuroendovascular Therapy (JSNET) between 2005 and 2009. Medical information about the patients was anonymized and retrospectively registered via a website. Data from 32,608 patients were analyzed. The number of treated patients constantly increased from 5,040 in 2005 to 7,406 in 2009 and the rate of octogenarians increased from 7.0% in 2005 to 10.4% in 2009. The proportion of procedures remained relatively constant, but ratios of angioplasty slightly increased from 32.8% in 2005 to 33.7% in 2009. Procedural complications were associated more frequently with acute stroke (9.6%), ruptured aneurysms (7.4%), intracranial artery disease (ICAD) (5.4%), and arteriovenous malformation (AVM, 5.2%). The number of patients requiring neuroendovascular treatment in Japan is increasing and the outcomes of such therapy are clinically acceptable. Details of each type of treatment will be investigated in sub-analyses of the database.


PKA and PDE4D3 anchoring to AKAP9 provides distinct regulation of cAMP signals at the centrosome.
Previous work has shown that the protein kinase A (PKA)-regulated phosphodiesterase (PDE) 4D3 binds to A kinase-anchoring proteins (AKAPs). One such protein, AKAP9, localizes to the centrosome. In this paper, we investigate whether a PKA-PDE4D3-AKAP9 complex can generate spatial compartmentalization of cyclic adenosine monophosphate (cAMP) signaling at the centrosome. Real-time imaging of fluorescence resonance energy transfer reporters shows that centrosomal PDE4D3 modulated a dynamic microdomain within which cAMP concentration selectively changed over the cell cycle. AKAP9-anchored, centrosomal PKA showed a reduced activation threshold as a consequence of increased autophosphorylation of its regulatory subunit at S114. Finally, disruption of the centrosomal cAMP microdomain by local displacement of PDE4D3 impaired cell cycle progression as a result of accumulation of cells in prophase. Our findings describe a novel mechanism of PKA activity regulation that relies on binding to AKAPs and consequent modulation of the enzyme activation threshold rather than on overall changes in cAMP levels. Further, we provide for the first time direct evidence that control of cell cycle progression relies on unique regulation of centrosomal cAMP/PKA signals.


Measurement of LED chips using a large-area silicon photodiode.
We propose the output power measurement of bare-wafer/chip light-emitting diodes (LEDs) using a large-area silicon (Si) photodiode with a simple structure and high accuracy relative to the conventional partial flux measurement using an integrating sphere. To obtain the optical characteristics of the LED chips measured using the two methods, three-dimensional ray-trace simulations are used to perform the measurement deviations owing to the chip position offset or tilt angle. The ray-tracing simulation results demonstrate that the deviation of light remaining in the integrating sphere is approximately 65% for the vertical LED chip and 53% for the flip-chip LED chip if the measurement distance in partial flux method is set to be 5-40 mm. By contrast, the deviation of light hitting the photodiode is only 15% for the vertical LED chip and 23% for the flip-chip LED chip if the large-area Si photodiode is used to measure the output power with the same measurement distance. As a result, the large-area Si photodiode method practically reduces the output power measurement deviations of the bare-wafer/chip LED, so that a high-accuracy measurement can be achieved in the mass production of the bare-wafer/chip LED without the complicated integrating sphere structure.


Unsupervised Domain Adaptation From Axial to Short-Axis Multi-Slice Cardiac MR Images by Incorporating Pretrained Task Networks.
Anisotropic multi-slice Cardiac Magnetic Resonance (CMR) Images are conventionally acquired in patient-specific short-axis (SAX) orientation. In specific cardiovascular diseases that affect right ventricular (RV) morphology, acquisitions in standard axial (AX) orientation are preferred by some investigators, due to potential superiority in RV volume measurement for treatment planning. Unfortunately, due to the rare occurrence of these diseases, data in this domain is scarce. Recent research in deep learning-based methods mainly focused on SAX CMR images and they had proven to be very successful. In this work, we show that there is a considerable domain shift between AX and SAX images, and therefore, direct application of existing models yield sub-optimal results on AX samples. We propose a novel unsupervised domain adaptation approach, which uses task-related probabilities in an attention mechanism. Beyond that, cycle consistency is imposed on the learned patient-individual 3D rigid transformation to improve stability when automatically re-sampling the AX images to SAX orientations. The network was trained on 122 registered 3D AX-SAX CMR volume pairs from a multi-centric patient cohort. A mean 3D Dice of 0.86 ± 0.06 for the left ventricle, 0.65 ± 0.08 for the myocardium, and 0.77 ± 0.10 for the right ventricle could be achieved. This is an improvement of 25% in Dice for RV in comparison to direct application on axial slices. To conclude, our pre-trained task module has neither seen CMR images nor labels from the target domain, but is able to segment them after the domain gap is reduced. Code: https://github.com/Cardio-AI/3d-mri-domain-adaptation.


Quantification of cysteinyl S-nitrosylation by fluorescence in unbiased proteomic studies.
Cysteinyl S-nitrosylation has emerged as an important post-translational modification affecting protein function in health and disease. Great emphasis has been placed on global, unbiased quantification of S-nitrosylated proteins because of physiologic and oxidative stimuli. However, current strategies have been hampered by sample loss and altered protein electrophoretic mobility. Here, we describe a novel quantitative approach that uses accurate, sensitive fluorescence modification of cysteine S-nitrosylation that leaves electrophoretic mobility unaffected (SNOFlo) and introduce unique concepts for measuring changes in S-nitrosylation status relative to protein abundance. Its efficacy in defining the functional S-nitrosoproteome is demonstrated in two diverse biological applications: an in vivo rat hypoxia-ischemia/reperfusion model and antimicrobial S-nitrosoglutathione-driven transnitrosylation of an enteric microbial pathogen. The suitability of this approach for investigating endogenous S-nitrosylation is further demonstrated using Ingenuity Pathways analysis that identified nervous system and cellular development networks as the top two networks. Functional analysis of differentially S-nitrosylated proteins indicated their involvement in apoptosis, branching morphogenesis of axons, cortical neurons, and sympathetic neurites, neurogenesis, and calcium signaling. Major abundance changes were also observed for fibrillar proteins known to be stress-responsive in neurons and glia. Thus, both examples demonstrate the technique's power in confirming the widespread involvement of S-nitrosylation in hypoxia-ischemia/reperfusion injury and in antimicrobial host responses.


Copper-catalyzed asymmetric three-component borylstannation: enantioselective formation of C-Sn bond.
In summary, a first copper-catalyzed synthesis of α-aryl-β-borylstannane compounds was accomplished through three-component borylstannation of aryl-substituted alkenes. In the exploration of an asymmetric variant, chiral sulfinylphosphine ligands proved advantageous in controlling stereochemistry of B-Cu addition and in promoting transmetalation of enantioenriched alkyl-Cu species. The stereochemical outcome supported a sequential syn-borylcupration and configuration-retentive transmetalation mechanism. Moreover, α-chiral β-borylstannanes were easily transformed into a diverse array of secondary alkylstannanes and triarylethane with high enantiomeric purity. The applications of chiral sulfinylphosphine ligands to other tandem Cu-B addition reactions are currently under investigation in our group.


Clinical significance of conformity index and gradient index in patients undergoing stereotactic radiosurgery for a single metastatic tumor.
OBJECTIVEAlthough the conformity index (CI) and the gradient index (GI), which were proposed by Paddick and colleagues, are both logically considered to correlate with good posttreatment results after stereotactic radiosurgery (SRS), this hypothesis has not been confirmed clinically. The authors' aim was to reappraise whether high CI values correlate with reduced tumor progression rates, and whether low GI values correlate with reduced complication incidences.METHODSThis was an institutional review board-approved, retrospective cohort study conducted using a prospectively accumulated database including 3271 patients who underwent Gamma Knife SRS for brain metastases (BMs) during the 1998-2016 period. Among the 3271 patients, 925 with a single BM at the time of SRS (335 women and 590 men, mean age 66 [range 24-93] years) were studied. The mean/median CIs were 0.62/0.66 (interquartile range [IQR] 0.53-0.74, range 0.08-0.88) and the mean/median GIs were 3.20/3.09 (IQR 2.83-3.39, range 2.27-11.4).RESULTSSRS-related complications occurred in 38 patients (4.1%), with a median post-SRS interval of 11.5 (IQR 6.0-25.8, maximum 118.0) months. Cumulative incidences of post-SRS complications determined by a competing risk analysis were 2.2%, 3.2%, 3.6%, 3.8%, and 3.9% at the 12th, 24th, 36th, 48th, and 60th post-SRS month, respectively. Multivariable analyses showed that only two clinical factors (i.e., peripheral doses and brain volume receiving ≥ 12 Gy) correlated with complication rates. However, neither CIs nor GIs impacted the incidences of complications. Among the 925 patients, post-SRS MRI was performed at least once in 716 of them, who were thus eligible for local progression evaluation. Among these 716 patients, local progression was confirmed in 96 (13.4%), with a median post-SRS interval of 10.8 (IQR 6.7-19.5, maximum 59.8) months. Cumulative incidences of local progression determined by a competing risk analysis were 7.7%, 12.6%, 14.2%, 14.8%, and 15.3% at the 12th, 24th, 36th, 48th, and 60th post-SRS month, respectively. Multivariable analyses showed neurological symptoms, extracerebral metastases, repeat SRS, and CIs to correlate with incidences of local progression, whereas GIs had no impact on local tumor progression. Particularly, cumulative incidences of local progression were significantly lower in patients with CIs < 0.65 than in those with CIs ≥ 0.65 (adjusted hazard ratio 1.870, 95% confidence interval 1.299-2.843; p = 0.0034).CONCLUSIONSTo the authors' knowledge, this is the first analysis to focus on the clinical significance of CI and GI based on a large series of patients with BM. Contrary to the majority opinion that dose planning with higher CI and lower GI results in good post-SRS outcomes (i.e., low local progression rates and minimal complications), this study clearly showed that the lower the CIs were, the lower the local progression rates were, and that the GI did not impact complication rates.


Immunohistochemical study of six cases of Taylor's type focal cortical dysplasia: correlation with electroclinical data.
Cortical specimens from six patients operated on for drug-resistant epilepsy diagnosed as Taylor's type focal cortical dysplasia were submitted to neuropathological and immunohistochemical studies. All patients were submitted to presurgical investigations including clinical and neuropsychological evaluations, EEG/video telemetry of ictal and interictal events, magnetic resonance imaging, and ictal and interictal single-photon emission computed tomography (SPECT). Recordings from electrocorticography (ECoG) were obtained in four cases and from subdural electrode implantation in two. Postsurgical follow-up was assessed according to Engel's score. Immunohistochemistry (IHC) was processed for parvalbumin (PV), calbindin D28-K (CB), nonphosphorylated neurofilaments (SMI-311), glial fibrillary acidic protein (GFAP) in all cases. We found continuous/quasi-continuous spikes and sharp-wave patterns in three cases and frequent repetitive bursting of polyspikes and ECoG seizures in two cases. Every patient showed cortical dyslamination, abnormal and giant neurons, and balloon cells. GFAP immunoreactivity was found in astrocytes and some balloon cells that were less intensely stained. Nonphosphorylated neurofilaments SMI-311 immunoreactivity was found in normal and giant neurons and in some balloon cells, making visible thin neuropils. PV immunoreactivity was present in normal interneurons and in fibers in layers IV-V. PV-negative balloon cells were surrounded by abundant PV-positive fibers. CB immunoreactivity was found mostly in interneurons in layers II-III. Our research is inconclusive. More cases should be investigated, and we must draw more accurate anatomic correlations between the ECoG recordings and surgical specimens studied with IHC.


Reproducibility of facial soft tissue landmarks on 3D laser-scanned facial images.
The three-dimensional (3D) measuring technology is useful to inspect facial shape in three planes of space (X, Y, and Z). Recent work has been directed to analyse craniofacial morphology using facial soft tissue landmarks to identify facial differences among population. The reproducibility of facial landmarks is almost necessary to ensure accurate 3D facial measurements. The aim of this study is to assess the reproducibility of facial soft tissue landmarks using laser-scan 3D imaging technology. Facial landmarks were assessed for 30 15(1/2)-year-old British-Caucasian children (15 males and 15 females). The sample was recruited from the Avon Longitudinal Study of Parents and Children (ALSPAC). The 3D facial images were acquired for each subject using two high-resolution Konica/Minolta laser scanners. Twenty-one facial landmarks (63 X, Y, and Z coordinates) were identified and recorded on each 3D facial image by two examiners. The reproducibility of landmarks identification at 2-week interval was assessed for one of the examiners (intra-examiner). In addition, the reproducibility of landmarks was assessed between the two examiners (inter-examiner). Using Bland-Altman plots, both intra- and inter-examiner assessments had evaluated landmarks reproducibility in three dimensions for the sample divided by gender. The reproducibility of the 3D-coordinates for each landmark was considered under three categories (< 0.5 mm, < 1 mm, and >1 mm) for both intra- and inter-examiner reproducibility assessments. The distribution of coordinates at the three levels of reproducibility show the following percentages: intra-examiner: < 0.5 mm (38%), < 1 mm (51%), >1 mm (11%); inter-examiner: < 0.5 mm (35%), < 1 mm (48%), >1 mm (17%). Generally, 10 landmarks were reproducible to less than 1 mm for both intra- and inter-examiner reproducibility assessments. The Labiale Superius was the most reproducible and Palebrale Superius was the least reproducible landmark. Some landmarks showed greater reliability in certain planes of space; the Glabella was more reliable in the Z than the Y axis. Gender differences were found; Subnasale was more reproducible in the Y-axis in males compared with females. The reproducibility of facial landmarks should be considered in the three planes of space. The majority of X-Y-Z coordinates taken to the 21 facial landmarks were reproducible to < 1 mm which is clinically acceptable. The accuracy of landmarks identification ranged from 0.39 to 1.49 mm. The reliability in identification depends on the clarity and definition of each landmark as well as gender characteristics. The different landmarks reproducibility should be considered when evaluating changes related to growth and healthcare interventions.


Sexual risk-taking at home and on holidays: the importance of context for the late application of condoms.
Several studies have examined condom use during 'holiday' sex but have not considered condom errors in this context. This study aims to identify factors associated with late application of condoms during participants' most recent vaginal intercourse at home and away from home (holidays or short breaks). Participants aged 19-30 years from a national Irish survey were recontacted (n=388; 51% men; mean age: 23.9 years). Telephone interviews regarding participants' most recent sex at home (n=362) and away from home (n=178) were conducted. A higher proportion reported condom use away from home (79% v. 62%), with a lower prevalence of late application (14% v. 24%). Pregnancy prevention as the primary motive for condom use increased the odds of late application at home (adjusted odds ratio (AOR): 4.56, 95% confidence interval (CI): 2.10-9.90) and away (AOR: 3.97, 95% CI: 1.36-11.59). A weak desire to use a condom also increased the likelihood of late application at home (AOR: 2.40, 95% CI: 1.03-5.62) and away (AOR: 11.18, 95% CI: 2.84-43.98). Subgroup analysis of those reporting both sexual events suggests that young adults take greater sexual risks with casual partners at home compared to away. The findings suggest that young adults take greater sexual risks at home than when away. Regardless of location, young adults are most likely to report late application when they have a weak desire to use a condom and when they use condoms primarily to prevent pregnancy.


Current parent education on infant feeding in the neonatal intensive care unit: the role of the occupational therapist.
The purpose of this study was to describe current trends in parent education on infant feeding in the neonatal intensive care unit (NICU) and to clarify the role of the occupational therapist in educating parents. Questionnaires were mailed to 190 neonatologists across the United States who were asked to forward it to a NICU occupational therapist. The questionnaire gathered descriptive information about the structure of parent education in the NICU, the role of the occupational therapist in providing parent education, and demographics about respondents and their NICUs. The response rate was 53% (n = 100). All 100 hospitals responding provided parent education in some form, and most included a variety of topics and teaching methods. Occupational therapists were on the NICU team at 74 of the hospitals and were identified third most frequently as a provider of parent education. The occupational therapists were most frequently identified as responsible for teaching about positioning, infant development, and infant states and cues and were highly involved in educating parents about feeding. Current parent education programs in NICUs are comprehensive in scope. Occupational therapists' role in educating parents about infant care and feeding consists of a focus on certain topics where occupational therapists have specialized skills and education. Occupational therapists are recognized by their NICU colleagues as providers of parent education, but this study suggests that the occupational therapists' role may not be clearly understood by other NICU professionals.


Antioxidant content of whole grain breakfast cereals, fruits and vegetables.
Considerable scientific evidence suggests that whole grains, as commonly consumed in the United States and Europe, reduce risk for chronic disease including cancer and heart disease. Whole grains provide a wide range of nutrients and phytochemicals that may work synergistically to optimize human health. Fruits and vegetables provide protection against age related diseases. It is believed their high content of antioxidant compounds is key to such protection. This research compares the antioxidant activity of whole grain, ready-to-eat (RTE) breakfast cereals to that of fruits and vegetables. Antioxidant activity was determined by dispersing finely ground samples in a 50% aqueous methanol solution of the stable free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). DPPH, which forms a deep purple solution, reacts with antioxidants and color loss at 515 nm correlates to antioxidant content, which is expressed as Trolox equivalents/100 grams (TE). Whole grain breakfast cereals analyzed in this study contained from 2,200-3,500 TE. By comparison, fruits generally ranged from 600-1,700 TE, with a high of 2,200 TE for red plums. Berries averaged 3,700 TE and. vegetables averaged 450 TE with a high of 1,400 TE for red cabbage. A 41 gram average serving of RTE breakfast cereal provides 1,120 TE, while an average 85 gram serving of vegetables or fruits provides 380 and 1,020 TE, respectively. Whole grain breakfast cereals, fruits and vegetables are all important dietary sources of antioxidants.


Beneficial effect of alpha-blocker on hemorheology in patients with essential hypertension.
To assess the effect of antihypertensive therapy on hemorheology in essential hypertension, blood viscosity and red blood cell deformability were examined in 45 patients with essential hypertension and 20 age-matched normotensive control subjects. Hypertensive patients were randomly assigned to monotherapy with five different antihypertensive drugs for 6 months and change of blood viscosity and red blood cell deformability were reexamined after the end of the monotherapy with each antihypertensive drug. Blood viscosity increased and red blood cell deformability decreased in hypertensive patients compared to normotensive control subjects. Monotherapy with each drug resulted in sufficient blood pressure control in all hypertensive patients. After the monotherapy with the alpha-blocker, terazosin, blood viscosity decreased significantly at shear rates from 22.5/sec to 450/sec, and red blood cell deformability, estimated by red blood cell filtration rate, increased by 15% (from 65 +/- 10 to 75 +/- 12 microL/sec, P < .05). The decrease in blood viscosity induced by alpha-blocker monotherapy may relate to an improvement of red blood cell deformability. It is possible that the treatment with alpha-blocker has a beneficial effect on the peripheral microcirculation due to an improvement of hemorheology in patients with essential hypertension.


Implications of a warming North Sea for the growth of haddock Melanogrammus aeglefinus.
The present study aimed firstly, to test for a temperature effect on North Sea haddock Melanogrammus aeglefinus growth and secondly, to develop a model that could be used to assess total length (L(T)) and mass (M)-at-age response to different temperature scenarios. The von Bertalanffy growth model was fitted on a cohort-by-cohort basis from 1970 to 2006. The asymptotic L(T) (L(∞)) was negatively correlated with temperature while the rate at which L(∞) is reached (K) was positively correlated with temperature. K was negatively correlated with density, whereas no effect on L(∞) was observed. These effects were incorporated into a von Bertalanffy model which was extended to include temperature and density as explanatory variables. Only the temperature variable was significant. Fitting the extended von Bertalanffy model revealed that L(∞) decreased while K increased with increasing temperature, resulting in up to a 40% loss of individual yield at older ages. The dramatic decline observed in the mean age at which 50% of the population becomes mature suggests that higher temperatures resulted in larger young M. aeglefinus that matured earlier and therefore reached a smaller maximum size. In a global warming context, the loss of individual yield observed at old ages is likely to reduce the fisheries yield for M. aeglefinus in the North Sea.


Comparison of clinical manifestations of HIV infection among women by risk group, CD4+ cell count, and HIV-1 plasma viral load. HER Study Group. HIV Epidemiology Research.
To compare the prevalence of HIV-related symptoms, physical examination findings, and hematologic variables among women whose risk for HIV is injection drug use since 1985 as opposed to sexual contact and to evaluate the influence of HIV plasma viral load and CD4+ cell count on clinical manifestations according to risk. Participants of the HIV Epidemiology Research Study (HERS; a multicenter, prospective, controlled study of HIV infection in women) were administered a risk behavior and symptom interview, underwent a physical examination, and received hematologic testing, including CD4+ cell counts done on study entry. Plasma HIV-1 viral loads were performed on stored frozen plasma using an ultrasensitive branched-DNA (b-DNA) signal amplification assay. CD4+ counts were categorized as <200 cells/microl, 200 to 499 cells/microl, or > or =500 cells/microl, and HIV viral loads were characterized in tertiles. Cross-sectional analysis was conducted on data available for 724 HIV-infected women: 387 had a history of intravenous drug use and 337 were infected through heterosexual contact. The median CD4+ count was 376 cells/microl; the median HIV-1 viral load was 1135 copies/ml; and 281 of 724 HIV-infected women (38.8%) had an undetectable HIV-1 viral load. In analyses adjusting for CD4+ cell level alone and for plasma viral load combined with CD4+ cell level, injection drug users (IDUs) were more likely than those infected through heterosexual contact to report a recent episode of memory loss and weight loss, but less likely to have recent episodes of genital herpes; to have enlarged livers and a body mass index (BMI) <24, and to have hematocrit levels <34% and platelet counts <150,000 cells/ml. After adjustment for CD4+ cell level and risk group, high and medium HIV-1 plasma viral load levels were associated with the presence of oral hairy leukoplakia on examination, and only the highest level of plasma viral load was associated with recent histories of fever and thrush, oral hairy leukoplakia, pseudomembranous candidiasis, and BMI <24 on examination, and hematocrit <34%. In this cohort of women, the distribution of HIV-1 plasma viral load was lower than that previously reported in populations of HIV-infected men. This study also shows some differences in frequency of signs, symptoms, and laboratory values between risk groups of HIV-infected women, but these results may be due to effects of injection drug use rather than HIV infection. Signs and symptoms identified as associated with increasing levels of viral load that were not different across risk groups suggest more direct association of these findings with HIV infection.


Bioinspired optofluidic FRET lasers via DNA scaffolds.
Optofluidic dye lasers hold great promise for adaptive photonic devices, compact and wavelength-tunable light sources, and micro total analysis systems. To date, however, nearly all those lasers are directly excited by tuning the pump laser into the gain medium absorption band. Here we demonstrate bioinspired optofluidic dye lasers excited by FRET, in which the donor-acceptor distance, ratio, and spatial configuration can be precisely controlled by DNA scaffolds. The characteristics of the FRET lasers such as spectrum, threshold, and energy conversion efficiency are reported. Through DNA scaffolds, nearly 100% energy transfer can be maintained regardless of the donor and acceptor concentration. As a result, efficient FRET lasing is achieved at an unusually low acceptor concentration of micromolar, over 1,000 times lower than that in conventional optofluidic dye lasers. The lasing threshold is on the order of μJ/mm(2). Various DNA scaffold FRET lasers are demonstrated to illustrate vast possibilities in optofluidic laser designs. Our work opens a door to many researches and applications such as intracavity bio/chemical sensing, biocontrolled photonic devices, and biophysics.


Birth-cohort changes in manic and depressive disorders in relatives of bipolar and schizoaffective patients.
Eight hundred twenty-three relatives of bipolar and schizoaffective patients were divided into groups of birth cohorts. The rates of bipolar, schizoaffective, and unipolar disorders were higher in the cohorts born after 1940 than in the cohorts born earlier. Life-table analysis of the cumulative hazard of illness in the different cohorts revealed significant differences among them, implying that the cumulative hazard of developing any affective disorder by a given age was greater in the post-1940 cohorts. When these data are combined with other reports, an ominous trend may be present, leading to an increase in prevalence of a broad spectrum of familial affective disorders in the coming decades.


Adjustment and social support at work early after breast cancer surgery and its associations with sickness absence.
As half of the women with breast cancer are of working ages and usually survive, knowledge is needed on how to support them early regarding work-related problems caused by treatments. Most previous studies have focused on individual and disease-related factors, whereas few have focused on work-related factors such as work adjustment and social support. The aim of this study was to investigate received and perceived social support from supervisor and colleagues as well as work adjustments, and their associations with sickness absence, among women who recently had had breast cancer surgery. Inclusion criteria were as follows: women aged 20-63 years, living in Stockholm County, treated surgically for a first diagnosis of breast cancer, literate in Swedish, without pre-surgical chemotherapy or known distant metastases. Included in the study were 605 women who worked at diagnosis and that had answered a questionnaire within eight weeks of inclusion. Descriptive statistics, univariate, and multivariable logistic regression analyses were applied to estimate odds ratios (OR) with 95% confidence intervals (CI) for the likelihood of being sickness absent. Most women perceived and received social support and work adjustment after breast cancer surgery. Low adjustment (OR = 2.14; 95% CI, 1.45-3.18) and less social support (OR = 1.80; 95% CI, 1.16-2.78) were significantly associated with being sickness absent. Adjusting for sociodemographics, strenuous work posture, and treatment did not attenuate these associations. Adjustment at work and social support from employer are associated with sickness absence and needs to be explored in discussions on return to work after breast cancer surgery.


Comparison of subjective, pharmacokinetic, and physiological effects of marijuana smoked as joints and blunts.
Recent increases in marijuana smoking among the young adult population have been accompanied by the popularization of smoking marijuana as blunts instead of as joints. Blunts consist of marijuana wrapped in tobacco leaves, whereas joints consist of marijuana wrapped in cigarette paper. To date, the effects of marijuana smoked as joints and blunts have not been systematically compared. The current within-subject, randomized, double-blind, placebo-controlled study sought to directly compare the subjective, physiological, and pharmacokinetic effects of marijuana smoked by these two methods. Marijuana blunt smokers (12 women and 12 men) were recruited and participated in a 6-session outpatient study. Participants were blindfolded and smoked three puffs from either a blunt or a joint containing marijuana with varying Delta(9)-tetrahydrocannabinol (THC) concentrations (0.0, 1.8, and 3.6%). Subjective, physiological (heart rate, blood pressure, and carbon monoxide levels) and pharmacokinetic effects (plasma THC concentration) were monitored before and at specified time points for 3h after smoking. Joints produced greater increases in plasma THC and subjective ratings of marijuana intoxication, strength, and quality compared to blunts, and these effects were more pronounced in women compared to men. However, blunts produced equivalent increases in heart rate and higher carbon monoxide levels than joints, despite producing lower levels of plasma THC. These findings demonstrate that smoking marijuana in a tobacco leaf may increase the risks of marijuana use by enhancing carbon monoxide exposure and increasing heart rate compared to joints.


Blood-brain barrier breakdown after embolic stroke in rats occurs without ultrastructural evidence for disrupting tight junctions.
The term blood-brain barrier (BBB) relates to the ability of cerebral vessels to hold back hydrophilic and large molecules from entering the brain, thereby crucially contributing to brain homeostasis. In fact, experimental opening of endothelial tight junctions causes a breakdown of the BBB evidenced as for instance by albumin leakage. This and similar observations led to the conclusion that BBB breakdown is predominantly mediated by damage to tight junction complexes, but evidentiary ultrastructural data are rare. Since functional deficits of the BBB contribute to an increased risk of hemorrhagic transformation and brain edema after stroke, which both critically impact on the clinical outcome, we studied the mechanism of BBB breakdown using an embolic model of focal cerebral ischemia in Wistar rats to closely mimic the essential human pathophysiology. Ischemia-induced BBB breakdown was detected using intravenous injection of FITC-albumin and tight junctions in areas of FITC-albumin extravasation were subsequently studied using fluorescence and electron microscopy. Against our expectation, 25 hours after ischemia induction the morphology of tight junction complexes (identified ultrastructurally and using antibodies against the transcellular proteins occludin and claudin-5) appeared to be regularly maintained in regions where FITC-albumin massively leaked into the neuropil. Furthermore, occludin signals along pan-laminin-labeled vessels in the affected hemisphere equaled the non-affected contralateral side (ratio: 0.966 vs. 0.963; P = 0.500). Additional ultrastructural analyses at 5 and 25 h after ischemia induction clearly indicated FITC-albumin extravasation around vessels with intact tight junctions, while the endothelium exhibited enhanced transendothelial vesicle trafficking and signs of degeneration. Thus, BBB breakdown and leakage of FITC-albumin cannot be correlated with staining patterns for common tight junction proteins alone. Understanding the mechanisms causing functional endothelial alterations and endothelial damage is likely to provide novel protective targets in stroke.


The Current Recommended Vitamin D Intake Guideline for Diet and Supplements During Pregnancy Is Not Adequate to Achieve Vitamin D Sufficiency for Most Pregnant Women.
The aims of this study were to determine if pregnant women consumed the recommended vitamin D through diet alone or through diet and supplements, and if they achieved the current reference range vitamin D status when their reported dietary intake met the current recommendations. Data and banked blood samples collected in second trimester from a subset of 537 women in the APrON (Alberta Pregnant Outcomes and Nutrition) study cohort were examined. Frozen collected plasma were assayed using LC-MS/MS (liquid chromatography-tandem mass spectrometry) to determine 25(OH)D2, 25(OH)D3, 3-epi-25(OH)D3 concentrations. Dietary data were obtained from questionnaires including a Supplement Intake Questionnaire and a 24-hour recall of the previous day's diet. Participants were 87% Caucasian; mean (SD) age of 31.3 (4.3); BMI 25.8 (4.7); 58% were primiparous; 90% had education beyond high school; 80% had a family income higher than CAN $70,000/year. 25(OH)D2, 25(OH)D3, and 3-epi-25(OH)D3) were identified in all of the 537 plasma samples;3-epi-25(OH)D3 contributed 5% of the total vitamin D. The median (IQR) total 25(OH)D (D2+D3) was 92.7 (30.4) nmol/L and 20% of women had 25(OH)D concentration < 75 nmol/L. The median (IQR) reported vitamin D intake from diet and supplements was 600 (472) IU/day. There was a significant relationship between maternal reported dietary vitamin D intake (diet and supplement) and 25(OH)D and 3-epi-25(OH)D3 concentrations in an adjusted linear regression model. We demonstrated the current RDA (600 IU/ day) may not be adequate to achieve vitamin D status >75 nmol/L in some pregnant women who are residing in higher latitudes (Calgary, 51°N) in Alberta, Canada and the current vitamin D recommendations for Canadian pregnant women need to be re-evaluated.