Change in vascular access and mortality in maintenance hemodialysis patients.
We hypothesized that a change from central venous catheters to a fistula or graft would improve short-term mortality risk in maintenance hemodialysis patients. Prospective observational study. All maintenance in-center hemodialysis patients treated in Fresenius Medical Care, North America legacy facilities alive on January 1, 2007 with baseline laboratory data from December 2006. Access type (fistula, catheter, or graft), determined on December 31, 2006, and monthly thereafter. Conversion from a catheter to a fistula or graft during the 4-month period from January 1 to April 30, 2007. Mortality was tracked from May 1, 2007, to December 31, 2007. Standard and time-dependent Cox models were used to determine hazard risks (HRs) of death with and without adjustment for case-mix and laboratory values. At baseline, 79,545 patients had 43% fistulas, 29% catheters, and 27% grafts. Mean age was 62 +/- 15 years, 54% were men, 51% were white, and 53% had diabetes. Compared with fistulas, unadjusted HRs of death were higher for grafts (1.22) and catheters (1.76; P < 0.001). In adjusted models, overall risk for grafts was decreased to 1.05 (95% limits, 1.003-1.100; P < 0.05) and approached that for fistulas consistently across multiple strata. Compared with patients who continued using a catheter, those who converted to either a graft or fistula had an HR of 0.69, whereas those who converted from a graft or fistula to a catheter had increased HRs to 2.12 (both P < 0.001). Similar trends were observed in the subset of incident patients (vintage < 90 days at study onset). Observational design with residual confounding from unmeasured patient, facility, and treatment-related factors. Catheters have the worst associated mortality risk. Changing from a catheter to a fistula or graft is associated with significantly improved survival. The risk for grafts approached that of fistulas, providing an alternative to prolonged catheter exposure and potentially less hazardous "bridge" toward a fistula.


Antigen-induced reduction in mast cell and basophil functional responses due to reduced Syk protein levels.
The high-affinity IgE receptor, FcepsilonRI, is unresponsive on mast cells and basophils from people in several populations through an unknown mechanism. Similarly, FcepsilonRI-positive basophils from 'nonreleasers' are IgE-unresponsive and are deficient in the tyrosine kinase Syk. To test the hypothesis that cross-linking FcepsilonRI on mast cells and basophils leads to FcepsilonRI nonresponsiveness through reduction in Syk protein levels. Human mast cells and basophils were used to determine if FcepsilonRI hyporesponsiveness correlated with reduced Syk levels. It is shown that suboptimal antigen challenge, that did not lead to significant mediator release, induced nonresponsiveness and correlated with reduced Syk. Other IgE-associated signaling molecules were unaffected by the same treatment. The ability of IgE-unresponsive mast cells to regain FcepsilonRI responsiveness is paralleled by increased cellular Syk levels in vitro. The reduction of Syk levels with suboptimal antigen concentrations was calcium independent and mediated through a proteasome-dependent mechanism. These findings confirm and extend our knowledge about a novel regulatory mechanism for maintaining FcepsilonRI in a quiescent state. This mechanism may also explain why low concentrations of allergen given to patients during allergen immunotherapy induce FcepsilonRI nonresponsiveness and therapeutic benefit without inducing systemic anaphylaxis.


Dynamic competition for hexon binding between core protein VII and lytic protein VI promotes adenovirus maturation and entry.
Adenovirus minor coat protein VI contains a membrane-disrupting peptide that is inactive when VI is bound to hexon trimers. Protein VI must be released during entry to ensure endosome escape. Hexon:VI stoichiometry has been uncertain, and only fragments of VI have been identified in the virion structure. Recent findings suggest an unexpected relationship between VI and the major core protein, VII. According to the high-resolution structure of the mature virion, VI and VII may compete for the same binding site in hexon; and noninfectious human adenovirus type 5 particles assembled in the absence of VII (Ad5-VII-) are deficient in proteolytic maturation of protein VI and endosome escape. Here we show that Ad5-VII- particles are trapped in the endosome because they fail to increase VI exposure during entry. This failure was not due to increased particle stability, because capsid disruption happened at lower thermal or mechanical stress in Ad5-VII- compared to wild-type (Ad5-wt) particles. Cryoelectron microscopy difference maps indicated that VII can occupy the same binding pocket as VI in all hexon monomers, strongly arguing for binding competition. In the Ad5-VII- map, density corresponding to the immature amino-terminal region of VI indicates that in the absence of VII the lytic peptide is trapped inside the hexon cavity, and clarifies the hexon:VI stoichiometry conundrum. We propose a model where dynamic competition between proteins VI and VII for hexon binding facilitates the complete maturation of VI, and is responsible for releasing the lytic protein from the hexon cavity during entry and stepwise uncoating.


Long jumpers with and without a transtibial amputation have different three-dimensional centre of mass and joint take-off step kinematics.
Long jumpers with below the knee amputation (BKA) have achieved remarkable performances, yet the underlying biomechanics resulting in these jump distances are unknown. We measured three-dimensional motion and used multi-segment modelling to quantify and compare the centre of mass (COM) and joint kinematics of three long jumpers with BKA and seven non-amputee long jumpers during the take-off step of the long jump. Despite having the same jump distances, athletes with BKA, who used their affected leg for the take-off step, had lower sagittal plane hip and knee joint range of motion and positioned their affected leg more laterally relative to the COM compared to non-amputee athletes. Athletes with BKA had a longer compression phase and greater downward movement of their COM, suggesting that their affected leg (lever) was less rigid compared to the biological leg of non-amputees. Thus, athletes with BKA used a different kinematic mechanism to redirect horizontal to vertical velocity compared to non-amputee athletes. The specific movement patterns of athletes with BKA during the take-off step were constrained by the mechanical properties of the prosthesis. These results provide a basis for coaches and athletes to develop training protocols that improve performance and inform the design of future prostheses.


Prioritizing Variants in Complete Hereditary Breast and Ovarian Cancer Genes in Patients Lacking Known BRCA Mutations.
BRCA1 and BRCA2 testing for hereditary breast and ovarian cancer (HBOC) does not identify all pathogenic variants. Sequencing of 20 complete genes in HBOC patients with uninformative test results (N = 287), including noncoding and flanking sequences of ATM, BARD1, BRCA1, BRCA2, CDH1, CHEK2, EPCAM, MLH1, MRE11A, MSH2, MSH6, MUTYH, NBN, PALB2, PMS2, PTEN, RAD51B, STK11, TP53, and XRCC2, identified 38,372 unique variants. We apply information theory (IT) to predict and prioritize noncoding variants of uncertain significance in regulatory, coding, and intronic regions based on changes in binding sites in these genes. Besides mRNA splicing, IT provides a common framework to evaluate potential affinity changes in transcription factor (TFBSs), splicing regulatory (SRBSs), and RNA-binding protein (RBBSs) binding sites following mutation. We prioritized variants affecting the strengths of 10 splice sites (four natural, six cryptic), 148 SRBS, 36 TFBS, and 31 RBBS. Three variants were also prioritized based on their predicted effects on mRNA secondary (2°) structure and 17 for pseudoexon activation. Additionally, four frameshift, two in-frame deletions, and five stop-gain mutations were identified. When combined with pedigree information, complete gene sequence analysis can focus attention on a limited set of variants in a wide spectrum of functional mutation types for downstream functional and co-segregation analysis.


Interventions for promoting booster seat use in four to eight year olds traveling in motor vehicles.
Public health and traffic safety agencies recommend use of booster seats in motor vehicles for children aged four to eight years, and various interventions have been implemented to increase their use by individuals who transport children in motor vehicles. There is little evidence regarding the effectiveness of these interventions, hence the need to examine what works and what does not. To assess the effectiveness of interventions intended to increase acquisition and use of booster seats in motor vehicles among four to eight year olds. We searched the Cochrane Injuries Group's Specialized Register, the Cochrane Central Register of Controlled Trials, MEDLINE (January 1966 to April 2005), EMBASE (1980 to April 2005), LILACS, Transport Research Databases (1988 to April 2005), Australian Transport Index (1976 to April 2005), additional databases and reference lists of relevant articles. We also contacted experts in the field. We included randomized and controlled before-and-after trials that investigated the effects of interventions to promote booster seat use. Two authors independently assessed trial quality and extracted data. Study authors were contacted for additional information. Five studies involving 3,070 individuals met the criteria for inclusion in the meta-analysis. All interventions for promoting use of booster seats among 4 to 8 year olds demonstrated a positive effect (relative risk (RR) 1.43; 95% confidence intervals (CI) 1.05 to 1.96). Incentives combined with education demonstrated a beneficial effect (RR 1.32, 95% CI 1.12 to 1.55; n = 1,898). Distribution of free booster seats combined with education also had a beneficial effect (RR 2.34; 95% CI 1.50 to 3.63; n = 380) as did education-only interventions (RR 1.32; 95% CI 1.16 to 1.49; n = 563). One study which evaluated enforcement of booster seat law met the criteria for inclusion in the meta-analysis, but demonstrated no marked beneficial effect. Available evidence suggests that interventions to increase use of booster seats among children age four to eight years are effective. Combining incentives (booster seat discount coupons or gift certificates) or distribution of free booster seats with education demonstrated marked beneficial outcomes for acquisition and use of booster seats for four to eight year olds. There is some evidence of beneficial effect of legislation on acquisition and use of booster seats but this was mainly from uncontrolled before-and-after studies, which did not meet the criteria for inclusion in the meta-analysis.


Cinnamaldehyde inhibits Candida albicans growth by causing apoptosis and its treatment on vulvovaginal candidiasis and oropharyngeal candidiasis.
The invasion of Candida albicans is one of the most common fungal infections seen in clinical practice, and serious drug resistance has been reported in recent years. Therefore, new anti-C. albicans drugs must be introduced. In this research, it was demonstrated that cinnamaldehyde (CA) shows strong antimicrobial activity, with 0.26 mg/mL CA being the minimum inhibitory concentration to manage C. albicans. Extraordinarily, we detected that CA accumulated the intracellular reactive oxygen species (ROS) and enhanced the calcium concentration in the cytoplasm and mitochondria through flow cytometry. In addition, we observed that C. albicans cells released Cytochrome c from the mitochondria to the cytoplasm, depolarized the mitochondrial membrane potential, and activated the metacaspase when exposed to 0.065, 0.13, 0.26, and 0.52 mg/mL CA. Furthermore, to confirm that CA introduces the C. albicans apoptosis, we discovered that when the phosphatidylserine was exposed, DNA damage and chromatin condensation occurred, which were detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and 4',6-diamidino-2-phenylindole (DAPI) staining. Finally, demonstrations of phenotype investigation, colony-forming unit (CFU) counts, and periodic acid-Schiff (PAS) staining were conducted to prove that CA possessed the ability to treat oropharyngeal candidiasis (OPC) and vulvovaginal candidiasis (VVC). From the above, our research indicates that CA is a promising antifungal candidate when applied to C. albicans infections.


Genetic differences in the frequency of the hinge variants of porcine IgA is breed dependent.
The distribution of the IgA(a) and IgA(b) alleles of porcine IgA in over 160 randomly-selected animals revealed an abundance of heterozygotes but only two b/b homozygotes. Since the IgA(b) allotype is a splice site mutant lacking two-thirds of the hinge, this study tests the hypothesis that pigs with this genotype may be at a selective disadvantage while heterozygous individuals may be at some advantage. This hypothesis was tested by collecting data on 374 animals of known breed and often parentage. We show here that when breed was not considered, young animals of known parentage had genotypic frequencies identical to that expected for Mendelian alleles but that a/b heterozygotes were overrepresented in adults. However, when analyzed with regard to breed, a very strong association between breed and the frequency of the IgA(a) and IgA(b) alleles was discovered. Meishan and NIH minipigs were homozygous for IgA while heterozygotes predominated in Berkshire, Chester White, Durocs, Hampshire and Landrace. Animals homozygous for IgA(b) were best represented in the White Cross line. We show here that this very strong breed dependency of IgA allotypy in swine can produce a sample bias that can explain why only two b/b homozygotes (1.3%) were found in the 160 randomly-selected samples since the original samples came from primarily Landrace and Yorkshire animals. The expected frequency of b/b homozygotes in these breeds would be <3%. Thus, the data presented here reject the hypothesis that swine homozygous for a trait that results in loss of two-thirds of the IgA hinge, are selected against and that heterozygotes are positively selected. Rather, the study shows that IgA(a) and IgA(b) appear to be simple, breed-dependent allotypic markers.


Diagnostic utility of anti-citrullinated protein antibody and its comparison with rheumatoid factor in rheumatoid arthritis.
To assess the diagnostic utility of anti-citrullinated protein antibody (anti-CCP) in rheumatoid arthritis (RA) and compare it with rheumatoid factor (RF). Analytical study. Section of Chemical Pathology, Department of Pathology and Microbiology and Medicine, the Aga Khan University, Karachi, from January to May 2010. A review of medical records of patients presenting to the clinics with complaints of muscular or joint pains and who were tested for their serum anti-CCP was done. Inclusion criteria were presence of clinical synovitis in at least one joint and an absence of alternative diagnosis. Patients with arthralgia alone or with missing acute phase reactants information were excluded. Scoring and classification of RA was done using the 2010 RA Classification Criteria by the American College of Rheumatology (ACR). Out of the 98 charts reviewed, ACR criteria showed 54 cases with RA. The mean age of the group was 46 ± 15 years, 82.7% being females. High titers of anti-CCP corresponded with the ACR scores. The sensitivity and specificity of anti-CCP and RF reactivity for the diagnosis of RA were 54.7% and 95.5% versus 59.3% and 88.4% respectively. Anti-CCP is useful for the diagnosis of RA due to its higher specificity as compared to RF and can predict disease severity.


Tuning the emission of a water-soluble 3-hydroxyflavone derivative by host-guest complexation.
3-Hydroxyflavone derivatives have great potential as fluorescent probes for bio-labeling in aqueous medium. They were extensively studied in various organic solvents for the "excited state intramolecular proton transfer" process, but seldom addressed in aqueous solution due to the poor water solubility. Herein, an amphiphilic molecule bearing 3-hydroxyflavone and oligo(ethylene oxide) (denoted as 3HF-EO) was designed and synthesized. Different from the fluorescence in organic solvents, 3HF-EO in aqueous solution showed a remarkable single fluorescence emission, which is ascribed to the fluorescence of its anionic species. We found that the fluorescence intensity could be efficiently tuned via host-guest complexation. α-CD has little effect on the emission, while β-CD and γ-CD lead to enhanced and reduced emissions of 3HF-EO, respectively. The 1H NMR and 2D NOESY NMR spectra indicate that α-CD barely had any interaction with 3HF-EO, while β-CD and γ-CD formed complexes with one and two 3HF-EO molecules, respectively. These results provide a sound explanation for the modulated fluorescence intensity.


Audio-Visual, Visuo-Tactile and Audio-Tactile Correspondences in Preschoolers.
Interest in crossmodal correspondences has recently seen a renaissance thanks to numerous studies in human adults. Yet, still very little is known about crossmodal correspondences in children, particularly in sensory pairings other than audition and vision. In the current study, we investigated whether 4-5-year-old children match auditory pitch to the spatial motion of visual objects (audio-visual condition). In addition, we investigated whether this correspondence extends to touch, i.e., whether children also match auditory pitch to the spatial motion of touch (audio-tactile condition) and the spatial motion of visual objects to touch (visuo-tactile condition). In two experiments, two different groups of children were asked to indicate which of two stimuli fitted best with a centrally located third stimulus (Experiment 1), or to report whether two presented stimuli fitted together well (Experiment 2). We found sensitivity to the congruency of all of the sensory pairings only in Experiment 2, suggesting that only under specific circumstances can these correspondences be observed. Our results suggest that pitch-height correspondences for audio-visual and audio-tactile combinations may still be weak in preschool children, and speculate that this could be due to immature linguistic and auditory cues that are still developing at age five.


Reconstructing SALMFamide Neuropeptide Precursor Evolution in the Phylum Echinodermata: Ophiuroid and Crinoid Sequence Data Provide New Insights.
The SALMFamides are a family of neuropeptides that act as muscle relaxants in echinoderms. Analysis of genome/transcriptome sequence data from the sea urchin Strongylocentrotus purpuratus (Echinoidea), the sea cucumber Apostichopus japonicus (Holothuroidea), and the starfish Patiria miniata (Asteroidea) reveals that in each species there are two types of SALMFamide precursor: an L-type precursor comprising peptides with a C-terminal LxFamide-type motif and an F-type precursor solely or largely comprising peptides with a C-terminal FxFamide-type motif. Here, we have identified transcripts encoding SALMFamide precursors in the brittle star Ophionotus victoriae (Ophiuroidea) and the feather star Antedon mediterranea (Crinoidea). We have also identified SALMFamide precursors in other species belonging to each of the five echinoderm classes. As in S. purpuratus, A. japonicus, and P. miniata, in O. victoriae there is one L-type precursor and one F-type precursor. However, in A. mediterranea only a single SALMFamide precursor was found, comprising two peptides with a LxFamide-type motif, one with a FxFamide-type motif, five with a FxLamide-type motif, and four with a LxLamide-type motif. As crinoids are basal to the Echinozoa (Holothuroidea + Echinoidea) and Asterozoa (Asteroidea + Ophiuroidea) in echinoderm phylogeny, one model of SALMFamide precursor evolution would be that ancestrally there was a single SALMFamide gene encoding a variety of SALMFamides (as in crinoids), which duplicated in a common ancestor of the Echinozoa and Asterozoa and then specialized to encode L-type SALMFamides or F-type SALMFamides. Alternatively, a second SALMFamide precursor may remain to be discovered or may have been lost in crinoids. Further insights will be obtained if SALMFamide receptors are identified, which would provide a molecular basis for experimental analysis of the functional significance of the "cocktails" of SALMFamides that exist in echinoderms.


Expression and intracellular transport of microvillus membrane hydrolases in human intestinal epithelial cells.
A panel of monoclonal antibodies was produced against purified microvillus membranes of human small intestinal enterocytes. By means of these probes three disaccharidases (sucrase-isomaltase, lactase-phlorizin hydrolase, and maltase-glucoamylase) and four peptidases (aminopeptidase N, dipeptidylpeptidase IV, angiotension I-converting enzyme, and p-aminobenzoic acid peptide hydrolase) were successfully identified as individual entities by SDS PAGE and localized in the microvillus border of the enterocytes by immunofluorescence microscopy. The antibodies were used to study the expression of small intestinal hydrolases in the colonic adenocarcinoma cell line Caco 2. This cell line was found to express sucrase-isomaltase, lactase-phlorizin hydrolase, aminopeptidase N, and dipeptidylpeptidase IV, but not the other three enzymes. Pulse-chase studies with [35S]methionine and analysis by subunit-specific monoclonal antibodies revealed that sucrase-isomaltase was synthesized and persisted as a single-chain protein comprising both subunits. Similarly, lactase-phlorizin hydrolase was synthesized as a large precursor about twice the size of the lactase subunits found in the human intestine. Aminopeptidase N and dipeptidylpeptidase IV, known to be dimeric enzymes in most mammals, were synthesized as monomers. Transport from the rough endoplasmic reticulum to the trans-Golgi apparatus was considerably faster for the peptidases than for the disaccharidases, as probed by endoglycosidase H sensitivity. These results suggest that the major disaccharidases share a common biosynthetic mechanism that differs from that for peptidases. Furthermore, the data indicate that the transport of microvillus membrane proteins to and through the Golgi apparatus is a selective process that may be mediated by transport receptors.


Clinical Practice Guideline for Endoscopic Resection of Early Gastrointestinal Cancer.
Although surgery was the standard treatment for early gastrointestinal cancers, endoscopic resection is now a standard treatment for early gastrointestinal cancers without regional lymph node metastasis. High-definition white light endoscopy, chromoendoscopy, and image-enhanced endoscopy such as narrow band imaging are performed to assess the edge and depth of early gastrointestinal cancers for delineation of resection boundaries and prediction of the possibility of lymph node metastasis before the decision of endoscopic resection. Endoscopic mucosal resection and/or endoscopic submucosal dissection can be performed to remove early gastrointestinal cancers completely by en bloc fashion. Histopathological evaluation should be carefully made to investigate the presence of risk factors for lymph node metastasis such as depth of cancer invasion and lymphovascular invasion. Additional treatment such as radical surgery with regional lymphadenectomy should be considered if the endoscopically resected specimen shows risk factors for lymph node metastasis. This is the first Korean clinical practice guideline for endoscopic resection of early gastrointestinal cancer. This guideline was developed by using mainly de novo methods and encompasses endoscopic management of superficial esophageal squamous cell carcinoma, early gastric cancer, and early colorectal cancer. This guideline will be revised as new data on early gastrointestinal cancer are collected.


Tandem repeat regions within the Burkholderia pseudomallei genome and their application for high resolution genotyping.
The facultative, intracellular bacterium Burkholderia pseudomallei is the causative agent of melioidosis, a serious infectious disease of humans and animals. We identified and categorized tandem repeat arrays and their distribution throughout the genome of B. pseudomallei strain K96243 in order to develop a genetic typing method for B. pseudomallei. We then screened 104 of the potentially polymorphic loci across a diverse panel of 31 isolates including B. pseudomallei, B. mallei and B. thailandensis in order to identify loci with varying degrees of polymorphism. A subset of these tandem repeat arrays were subsequently developed into a multiple-locus VNTR analysis to examine 66 B. pseudomallei and 21 B. mallei isolates from around the world, as well as 95 lineages from a serial transfer experiment encompassing ~18,000 generations. B. pseudomallei contains a preponderance of tandem repeat loci throughout its genome, many of which are duplicated elsewhere in the genome. The majority of these loci are composed of repeat motif lengths of 6 to 9 bp with 4 to 10 repeat units and are predominately located in intergenic regions of the genome. Across geographically diverse B. pseudomallei and B.mallei isolates, the 32 VNTR loci displayed between 7 and 28 alleles, with Nei's diversity values ranging from 0.47 and 0.94. Mutation rates for these loci are comparable (>10-5 per locus per generation) to that of the most diverse tandemly repeated regions found in other less diverse bacteria. The frequency, location and duplicate nature of tandemly repeated regions within the B. pseudomallei genome indicate that these tandem repeat regions may play a role in generating and maintaining adaptive genomic variation. Multiple-locus VNTR analysis revealed extensive diversity within the global isolate set containing B. pseudomallei and B. mallei, and it detected genotypic differences within clonal lineages of both species that were identical using previous typing methods. Given the health threat to humans and livestock and the potential for B. pseudomallei to be released intentionally, MLVA could prove to be an important tool for fine-scale epidemiological or forensic tracking of this increasingly important environmental pathogen.


Reliability and validity of the Chinese version of the HIV Treatment Adherence Self-Efficacy Scale in mainland China.
The aim of this study was to translate the HIV Treatment Adherence Self-Efficacy Scale (HIV-ASES) and evaluate its reliability and validity in people living with HIV (PLWHIV) in mainland China. The original HIV-ASES was translated from English into Chinese and adapted for cultural context according to cross-cultural adaptation guidelines. A cross-sectional survey investigating 1742 PLWHIV receiving antiretroviral treatment was conducted. Exploratory factor analysis with varimax rotation confirmed the presence of two factors with an accumulated contribution rate of 58.357%. Furthermore, confirmatory factor analysis revealed the model's goodness-of-fit index. The comparative fit index was greater than 0.9 and the overall efficacy of the model was satisfactory. The Cronbach α coefficient of the questionnaire was 0.876 (95% CI: 0.868-0.885), and the correlation coefficient of each item and the total was 0.536-0.660 ( P < 0.01). The Chinese version of the HIV-ASES has high reliability and validity; however, the individual item requires some adjustment.


Racial discrimination, post-traumatic stress and prescription drug problems among Aboriginal Canadians.
1) To examine associations between racial discrimination and drug problems among urban-based Aboriginal adults; and 2) to determine whether these associations are best explained by symptoms of psychological stress, distress or post-traumatic stress disorder (PTSD). Data were collected through in-person surveys with a community-based sample of Aboriginal adults (N = 372) living in a mid-sized city in western Canada in 2010. Associations were examined using bootstrapped linear regression models adjusted for confounders, with continuous prescription and illicit drug problem scores as outcomes. Mediation was examined using the cross-products of coefficients method. More than 80% of Aboriginal adults had experienced racial discrimination in the past year, with the majority reporting high levels in that period. Past-year discrimination was a risk factor for PTSD symptoms and prescription drug problems in models adjusted for confounders and other forms of psychological trauma. In mediation models, PTSD symptoms explained the association between discrimination and prescription drug problems; psychological stress and distress did not. PTSD symptoms also explained this association when the covariance between mediators was controlled. The results also indicate that participation in Aboriginal cultural traditions was associated with increased discrimination. Most efforts to address Aboriginal health inequities in Canada have focused on the role Aboriginal people play in these disparities. The current findings combine with others to call for an expanded focus. Non-Aboriginal Canadians may also play a role in the health inequities observed. The findings of this study suggest efforts to reduce discrimination experienced by Aboriginal adults in cities may reduce PTSD symptomology and prescription drug problems in these populations.


Transcriptomic responses of Atlantic salmon (Salmo salar) to environmental enrichment during juvenile rearing.
Captive rearing programs (hatcheries) are often used in conservation and management efforts for at-risk salmonid fish populations. However, hatcheries typically rear juveniles in environments that contrast starkly with natural conditions, which may lead to phenotypic and/or genetic changes that adversely affect the performance of juveniles upon their release to the wild. Environmental enrichment has been proposed as a mechanism to improve the efficacy of population restoration efforts from captive-rearing programs; in this study, we examine the influence of environmental enrichment during embryo and yolk-sac larval rearing on the transcriptome of Atlantic salmon (Salmo salar). Full siblings were reared in either a hatchery environment devoid of structure or an environment enriched with gravel substrate. At the end of endogenous feeding by juveniles, we examined patterns of gene transcript abundance in head tissues using the cGRASP-designed Agilent 4×44K microarray. Significance analysis of microarrays (SAM) indicated that 808 genes were differentially transcribed between the rearing environments and a total of 184 gene ontological (GO) terms were over- or under-represented in this gene list, several associated with mitosis/cell cycle and muscle and heart development. There were also pronounced differences among families in the degree of transcriptional response to rearing environment enrichment, suggesting that gene-by-environment effects, possibly related to parental origin, could influence the efficacy of enrichment interventions.


The Course of Neurocognitive Functioning and Prediction of Behavioral Outcome of ADHD Affected and Unaffected Siblings.
Longitudinal studies on the course of neurocognitive functioning of children with ADHD and their unaffected siblings are scarce. Also, it is unclear to what extent that course is related to ADHD outcomes. A carefully phenotyped large sample of 838 Caucasian participants (ADHD-combined type: n = 339, unaffected siblings: n = 271, controls: n = 228; mean age at baseline = 11.4 years, mean age at follow-up = 17.3 years, SD = 3.2) was used to investigate differences in the course of neurocognitive functioning of ADHD affected and unaffected siblings versus controls, and to investigate the relationship between neurocognitive change and ADHD outcomes. At baseline, an aggregated measure of overall neurocognitive functioning and eight neurocognitive measures of working memory, timing (speed/variability), motor control, and intelligence were investigated. Outcomes at follow-up were dimensional measures of ADHD symptom severity and the Kiddie-Global Assessment Scale (K-GAS) for overall functioning. At follow up, affected and unaffected siblings trended to, or fully caught up with performance levels of controls on four (44.4%) and five (55.6%) of the nine dependent variables, respectively. In contrast, performance in remaining key neurocognitive measures (i.e. verbal working memory, variability in responding) remained impaired at follow-up. Change in neurocognitive functioning was not related to ADHD outcomes. Our results question the etiological link between neurocognitive deficits and ADHD outcomes in adolescents and young adults.


Initiation of Bacillus spore germination by hydrostatic pressure: effect of temperature.
Suspensions of Bacillus cereus T, B. subtilis, and B. pumilus spores in water or potassium phosphate buffer were germinated by hydrostatic pressures of between 325 and 975 atm. Kinetics of germination at temperatures within the range of 25 to 44 degrees C were determined, and thermodynamic parameters were calculated. The optimum temperature for germination was dependent on pressure, species, suspending medium, and storage time after heat activation. Germination rates increased significantly with small increments of pressure, as indicated by high negative deltaV values of -230 +/- 5 cm3/mol for buffered B. subtilis (500 to 700 atm) and B. pumilus (500 atm) spores and -254 +/- 18 cm3/mol for aqueous B. subtilis (400 to 550 atm) spores at 40 degrees C and -612 +/- 41 cm3/mol for B. cereus (500 to 700 atm) spores at 25 degrees C. The ranges of thermodynamic constants calculated at 40 degrees C for buffered B. pumilus and B. subtilis spores at 500 and 600 atm and for aqueous B. subtilis spores at 500 atm were: Ea = 181,000 to 267,000 J/mol; deltaH = 178,000 to 264,000 J/mol; deltaG = 94,000 to 98,300 J/mol; deltaS = 264 to 544 J/mol per degree K. These values are consistent with the concept that the transformation of a dormant to a germinating spore induced by hydrostatic pressure involves either hydration or a reduction in the visocosity of the spore core and a conformational change of an enzyme.


Self-report prospective memory problems in people with stroke.
Prospective memory (PM) is a common problem which can limit performance of basic and instrumental activities of daily living in patients with stroke. This study compared self-report PM failures between older and younger people with stroke, examined differences in perceptions of PM failures between people with stroke and relatives, relationships between these PM failures and functional performance. A total of 105 patients with stroke, 65 relatives and 112 healthy controls were recruited. Both the patients with stroke and controls were further divided into an older (age > 55 years) and a younger (age ≤ 55 years) group. Data for patients with stroke and relatives were obtained via the Brief Assessment of Prospective Memory (BAPM), Basic Activity of Daily Living (BADL) related Modified Barthel Index (MBI) and Lawton Instrumental Activities of Daily Living (Lawton IADL) Scale. Healthy controls' data were also collected. The older stroke group had significantly higher BAPM total scores and BADL and IADL sub-scale scores than the younger group. Difference in perceptions of the patients' self-report of PM failures and their relatives' report was significant for the IADL sub-scale. Self-report of PM failure was significantly related to functional BADL and IADL measures. Results highlight the impact of PM failures in patients with stroke and their assessment, management and rehabilitation of these patients.


Adrenocortical function during prolonged critical illness and beyond: a prospective observational study.
For patients suffering from prolonged critical illness, it is unknown whether and when the hypothalamus-pituitary-adrenal axis alterations recover, and to what extent adrenocortical function parameters relate to sepsis/septic shock, to clinical need for glucocorticoid treatment, and to survival. Patients still in ICU on day 7 (N = 392) and 20 matched healthy subjects were included. Morning blood and 24-h urine were collected daily and cosyntropin tests (250 µg) performed weekly, repeated 1 week after ICU discharge on the regular ward. In all patients free of glucocorticoid treatment up until ICU day 28 (N = 347), plasma ACTH always remained low/normal, whereas free cortisol remained high (P ≤ 0.002) explained by reduced binding proteins (P ≤ 0.02) and suppressed cortisol breakdown (P ≤ 0.001). Beyond ICU day 28 (N = 64 long-stayers), plasma (free)cortisol was no longer elevated. One week after ICU discharge, plasma ACTH and (free)cortisol always rose to supra-normal levels (P ≤ 0.006), most pronounced in long-stayers. Long-stayers always showed low incremental total (P ≤ 0.001), but normal incremental free cortisol responses to weekly cosyntropin tests, explained by low cortisol plasma binding proteins. Sepsis/septic shock patients were not different from others, patients subsequently receiving glucocorticoids (N = 45) were not different from those who did not, and non-survivors were distinguishable from survivors only by higher (free)cortisol. Irrespective of sepsis/septic shock, need for glucocorticoids and survival, low cortisol plasma binding proteins and suppressed cortisol breakdown determine systemic (free)cortisol availability in prolonged critical illness, the latter no longer elevated beyond ICU day 28. The uniform rise in ACTH and cortisol to supra-normal levels 1 week after ICU discharge indicates recovery of a central adrenocortical suppression while in ICU. Low cortisol plasma binding invalidates the cosyntropin test.


Challenging the Cinderella Hypothesis: A New Model for the Role of the Motor Unit Recruitment Pattern in the Pathogenesis of Myofascial Pain Syndrome in Postural Muscles.
The energy crisis hypothesis, which is a widely accepted model for the pathogenesis of myofascial pain, has been corroborated by experimental observations. However, the nature of the insult leading to the energy crisis remains elusive. A commonly cited model for this insult is the Cinderella hypothesis, suggesting that hierarchical recruitment of motor units leads to a disproportional load on small units, thus driving them towards an energy crisis. New findings cast doubt on this model, showing that in postural muscles motor units are recruited in rotation, rather than in a hierarchical order, precluding the formation of the so-called Cinderella units. To explore the influence of common myofascial predisposing factors such as muscle load and muscle strength on the relaxation time of postural muscle motor units, assuming they are recruited in rotation. A stochastic model of a postural skeletal muscle was developed which integrates the energy crisis model and motor unit rotation patterns observed in postural muscles. Postulating that adequate relaxation time is essential for the energetic replenishment of motor units, we explored the influence of different parameters on the relaxation time of individual motor units under varying conditions of muscle loads and muscle strengths. The motor unit relaxation/contraction time ratio decreases with elevated muscle loads and with decreased total muscle strength. In a model of a postural muscle, in which motor units are recruited in rotation, common predisposing factors of myofascial pain, such as increased muscle load and decreased muscle force, lead to shortened motor unit relaxation periods.


Experimental observation of moiré angles in parallax barrier 3D displays.
Angles of visible moiré patterns are observed experimentally. Experiments were made across the angular range 0 - 90° in a wide range of parameters. Two kinds of clusterization were observed, ray and discrete. In rational cells (LCD pixels), the moiré patterns appear at a few fixed discrete angles. The list of preferable moiré-less angles is presented basing on the experimental data; preferable areas in the parameter space are found. The problem of minimization of the moiré effect is formulated as the Diophantine inequality with complex coefficients. The classification of moiré angles basing on the probability of the moiré effect can be practically useful.


Experimental Neutron Capture Rate Constraint Far from Stability.
Nuclear reactions where an exotic nucleus captures a neutron are critical for a wide variety of applications, from energy production and national security, to astrophysical processes, and nucleosynthesis. Neutron capture rates are well constrained near stable isotopes where experimental data are available; however, moving far from the valley of stability, uncertainties grow by orders of magnitude. This is due to the complete lack of experimental constraints, as the direct measurement of a neutron-capture reaction on a short-lived nucleus is extremely challenging. Here, we report on the first experimental extraction of a neutron capture reaction rate on ^{69}Ni, a nucleus that is five neutrons away from the last stable isotope of Ni. The implications of this measurement on nucleosynthesis around mass 70 are discussed, and the impact of similar future measurements on the understanding of the origin of the heavy elements in the cosmos is presented.


The spatiotemporal characteristics of elementary audiovisual speech and music processing in musically untrained subjects.
Previously, the EEG technique has been used to investigate the spatiotemporal properties of audiovisual (AV) processing by taking advantage of the violation of the "additive model", which is considered to be a very conservative approach. In the present work, we used a less conservative and novel approach than the criterion of superadditivity for estimating AV interactions. Hence, we estimated AV interaction patterns by comparing the responses to AV stimuli with the averaged responses to the unimodal visual and auditory stimuli in musically untrained subjects and by presenting syllables and piano tones coupled with flashlights. Our results suggest that the two AV objects elicited consistent interaction patterns within the time course of unisensory processing in the time range between 80 and 250ms post stimulus onset. The scalp topographies, as well as the source estimation approach we adopted, indicate that the first interaction pattern at around 100ms was partially driven by auditory-related cortical regions. Additionally, we found evidence for a second interaction pattern at around 200ms that was mainly associated with the responsiveness of extra-sensory brain regions. During this later processing stage, only the music condition was associated with putative responses that originated from auditory-related cortical fields. This study provides a novel approach to investigate the basic principles underlying elementary AV speech and music processing in subjects without formal musical education.


Retrospective economic analysis of the transfer of services from hospitals to the community: an application to an enhanced eye care service.
This research aims to evaluate the wider health system effects of the introduction of an intermediate-tier service for eye care. This research employs the Minor Eye Conditions Scheme (MECS), an intermediate-tier eye care service introduced in two London boroughs, Lewisham and Lambeth, in April 2013. Retrospective difference-in-differences analysis comparing changes over time in service use and costs between April 2011 and October 2014 in two commissioning areas that introduced an intermediate-tier service programme with changes in a neighbouring area that did not introduce the programme. MECS audit data; unit costs for MECS visits; volumes of first and follow-up outpatient attendances to hospital ophthalmology; the national schedule of reference costs. Volumes and costs of patients treated. In one intervention area (Lewisham), general practitioner (GP) referrals to hospital ophthalmology decreased differentially by 75.2% (95% CI -0.918% to -0.587%) for first attendances, and by 40.3% for follow-ups (95% CI -0.489% to -0.316%). GP referrals to hospital ophthalmology decreased differentially by 30.2% (95% CI -0.468% to -0.137%) for first attendances in the other intervention area (Lambeth). Costs increased by 3.1% in the comparison area between 2011/2012 and 2013/2014. Over the same period, costs increased by less (2.5%) in one intervention area and fell by 13.8% in the other intervention area. Intermediate-tier services based in the community could potentially reduce volumes of patients referred to hospitals by GPs and provide replacement services at lower unit costs.


Renoprotective approaches and strategies in acute kidney injury.
Acute kidney injury (AKI) is a major renal disease associated with high mortality rate and increasing prevalence. Decades of research have suggested numerous chemical and biological agents with beneficial effects in AKI. In addition, cell therapy and molecular targeting have been explored for reducing kidney tissue damage and promoting kidney repair or recovery from AKI. Mechanistically, these approaches may mitigate oxidative stress, inflammation, cell death, and mitochondrial and other organellar damage, or activate cytoprotective mechanisms such as autophagy and pro-survival factors. However, none of these findings has been successfully translated into clinical treatment of AKI. In this review, we analyze these findings and propose experimental strategies for the identification of renoprotective agents or methods with clinical potential. Moreover, we propose the consideration of combination therapy by targeting multiple targets in AKI.


Targeting Th17 Cells with Small Molecules and Small Interference RNA.
T helper 17 (Th17) cells play a central role in inflammatory and autoimmune diseases via the production of proinflammatory cytokines interleukin- (IL-) 17, IL-17F, and IL-22. Anti-IL-17 monoclonal antibodies show potent efficacy in psoriasis but poor effect in rheumatoid arthritis (RA) and Crohn's disease. Alternative agents targeting Th17 cells may be a better way to inhibit the development and function of Th17 cells than antibodies of blocking a single effector cytokine. Retinoic acid-related orphan receptor gamma t (RORγt) which acts as the master transcription factor of Th17 differentiation has been an attractive pharmacologic target for the treatment of Th17-mediated autoimmune disease. Recent progress in technology of chemical screen and engineering nucleic acid enable two new classes of therapeutics targeting RORγt. Chemical screen technology identified several small molecule specific inhibitors of RORγt from a small molecule library. Systematic evolution of ligands by exponential enrichment (SELEX) technology enabled target specific aptamers to be isolated from a random sequence oligonucleotide library. In this review, we highlight the development and therapeutic potential of small molecules inhibiting Th17 cells by targeting RORγt and aptamer mediated CD4(+) T cell specific delivery of small interference RNA against RORγt gene expression to inhibit pathogenic effector functions of Th17 lineage.


A moving blocker system for cone-beam computed tomography scatter correction.
Scatter contamination in cone-beam computed tomography (CBCT) degrades the image quality by introducing shading artifacts. A moving-blocker-based approach has been proposed to simultaneously estimate scatter and reconstruct the complete volume within field of view (FOV) from a single CBCT scan. Promising results were obtained from our simulation studies. In this work, the authors demonstrated experimentally the effectiveness of the moving-blocker-based scatter correction approach by implementing a moving blocker system on a LINAC on-board kV CBCT imaging system. A physical attenuator (i.e., "blocker") consisting of equal spaced lead strips was mounted on a linear actuator. A step motor connected to the actuator drove the blocker to move back and forth along gantry rotation axis during the CBCT acquisition. Scatter signal was estimated from the blocked region of imaging panel, and interpolated into the unblocked region. A statistics-based iterative image reconstruction algorithm was used to reconstruct CBCT images from unblocked projections after the scatter signal was subtracted. Experimental studies were performed on a Catphan phantom and an anthropomorphic pelvis phantom to evaluate performance of the moving blocker system. The scatter-induced shading artifacts were substantially reduced in the images acquired with the moving blocker system. CT number error in selected regions of interest reduced from 318 to 17 and from 239 to 10 for the Catphan phantom and pelvis phantom, respectively. The authors demonstrated experimentally that the moving blocker system can successfully estimate the scatter signal in projection data and obtain complete volumetric information within the FOV using a single scan.


Miniaturized tunable imaging system inspired by the human eye.
We demonstrate the combination of a tunable lens and tunable aperture in a compact imaging system whose structure is inspired by the human eye. The concept is based on innovative optical materials and tuning mechanisms including an optofluidic iris for a tunable aperture and an elastomer lens for focal length tuning. Wavefront and modulation transfer function analysis reveals a high imaging performance of both the individual elements and the complete system. Most significantly, the wavefront is not degraded (ΔPV<λ/10) when tuning the focal length (Δf=5.36%).


Treatment of Unstable Versus Stable Slipped Capital Femoral Epiphysis Using the Modified Dunn Procedure.
The modified Dunn procedure (open subcapital realignment via a surgical dislocation approach) has been shown to be a safe and effective way of treating acute, unstable slipped capital femoral epiphysis (SCFE). There is a paucity of literature comparing the modified Dunn procedure in stable SCFE. The purpose of this study was to compare acute, unstable versus chronic, stable SCFE managed with the modified Dunn procedure. A retrospective chart review was performed on 44 skeletally immature patients who underwent the modified Dunn procedure for SCFE. Patients were divided into stable or unstable based on clinical presentation and intraoperative findings. Demographics, radiographic measurements, and complications were recorded and compared. χ and t tests were used to compare variables. In total, 31 consecutive hips (29 patients) with acute, unstable slips, and 17 consecutive hips (15 patients) with chronic, stable slips were reviewed. Average age was 12.5 and 13.8 years for acute and chronic, respectively (P=0.05). Mean follow-up was 27.9 months (unstable) and 35.5 months (stable). Average postoperative Southwick angle was 14.2 degrees; (unstable) and 25.3 degrees (stable) (P=0.001). Greater trochanteric height averaged 6.2 mm below the center of the femoral head in the acute group and 6.2 mm above center in the chronic group (P<0.001). Average femoral neck length measured 34.1 mm (unstable) and 27.1 mm (stable) (P<0.001). Two patients (6%) developed avascular necrosis (AVN) in the unstable group, with 5 patients (29.4%) in the stable group (P=0.027). All patients with hip instability (N=3) developed AVN. Although both acute, unstable and chronic, stable SCFE can be successfully treated with the modified Dunn procedure, the complication rate is statistically higher in patients with stable SCFE, specifically both AVN rate and postoperative instability. In addition, it is more difficult to establish normal anatomic indexes with regard to greater trochanteric height and femoral neck length. This procedure has great utility in the correction of the anatomic deformity associated with SCFE, but should be used with caution in patients with chronic, stable SCFE. Level III-retrospective review.


3D Bioprinting of Breast Cancer Models for Drug Resistance Study.
Adipose-derived mesenchymal stem/stromal cells (ADMSC) are one of the major stromal cells in the breast cancer microenvironment that promote cancer progression. Previous studies on the effects of ADMSC on breast cancer metastasis and drug resistance, using two-dimensional (2D) cultures, remained inconclusive. In the present study, we compared cocultured ADMSC and human epidermal receptor 2 positive breast primary breast cancer cells (21PT) in 2D and three-dimensional (3D) cultures and then examined their response to doxorubicin (DOX). We examined 3D bioprinted constructs with breast cancer cells in the middle and ADMSC in the edge region, which were made by using dual hydrogel-based bioinks. We found that the percentage of cleaved Caspase-3 positive cells was significantly lower in the bioprinted constructs with ADMSC and 21PT than that in the cancer cell alone constructs, in response to low DOX dose. We further increased the thickness of the ADMSC layers to mimic the status of obesity and then examined the effect of ADMSC thickness on DOX resistance and lysyl oxidase (LOX) secretion. In the moderate and thick-layered ADMSC constructs, significantly more cells were stained negative for cleaved Caspase-3, indicating less apoptosis. Both ADMSC and 21PT intrinsically expressed LOX, regardless of changes in thickness or DOX administration. Notably, treatment with a LOX inhibitor significantly decreased the stiffness in the ADMSC region but did not affect the stiffness in the 21PT region. In addition, LOX inhibitor treatment enhanced DOX sensitivity of 21PT in the bioprinted constructs, as seen by a decrease in LOX secretion and downregulation of adenosine triphosphate-binding cassette transporter gene expression. Taken together, we demonstrate that 3D bioprinted these breast cancer models faithfully reproduce in vivo conditions and should provide better models for examining breast cancer biology and for screening for drug discoveries.


Biochip data normalization using multifunctional probes.
Using a biochip with stable probe functionalization and a detection system capable of real time measurements, it is demonstrated that acquired probe-target interaction data are more reproducible in time--on a given probe spot using sequential target runs--than in space, using many probe spot replicates on the biochip in one single parallel target run. To increase the biochip data precision, a normalization method that quantifies and corrects the surface inhomogeneity without the use of complex data post-processing has been developed. This simple and effective method is based on adding a common reactive group to all probes and quantifying the biochip response to a calibration target, thus quantifying the spatial heterogeneity in the biosensor responsiveness. The usefulness of such methodology, which can be easily generalized, is demonstrated in the model case of DNA:DNA interactions, using a surface plasmon resonance imaging system as the dynamical reader. The biochips are based on streptavidin biochemically functionalized gold films onto which biotinylated ssDNA probe sequences, related to cystic fibrosis genotyping, are spotted. This normalization method provides high gain in data precision and allows, in this example, unambiguous genotyping of SNP, including discrimination of the heterozygote case from the two homozygote cases.


Limnophyes guarani sp. n., a new hygropetric Orthocladiinae from southern Brazil (Diptera: Chironomidae).
A new species of Limnophyes Eaton collected in the Corvo Branco Mountains in Santa Catarina State is described and figured, based on male and female adults, larva and pupa. The species groups with L. griseata (Edwards) and L. bidumus Sæther as the adults have a pronounced humeral pit with lanceolate setae and an additional group of lanceolate setae just posterior to antepronotum. The adults are, however, distinctly smaller, have a lower AR and have more lanceolate setae in the group just posterior to antepronotum than the two other species. The pupa can easily be separated from the pupa of L. bidumus as the anal macroseta is longer than the anal lobe. The larva lives hygropetric on vertical rock surfaces. A key to male adults of Neotropical Limnophyes is given.


Efficacy of piroxicam plus cisplatin-loaded PLGA nanoparticles in inducing apoptosis in mesothelioma cells.
Combined treatment based on cisplatin-loaded Poly(D,L-lactic-co-glicolic)acid (PLGA) nanoparticles (NP-C) plus the NSAID piroxicam was used as novel treatment for mesothelioma to reduce side effects related to cisplatin toxicity. PLGA nanoparticles were prepared by double emulsion solvent evaporation method. Particle size, drug release profile and in vitro cellular uptake were characterized by TEM, DLS, LC/MS and fluorescence microscopy. MSTO-211H cell line was used to analyse NP-C biological efficacy by FACS and protein analysis. Cisplatin was encapsulated in 197 nm PLGA nanoparticles with 8.2% drug loading efficiency and 47% encapsulation efficiency. Cisplatin delivery from nanoparticles reaches 80% of total encapsulated drug in 14 days following a triphasic trend. PLGA nanoparticles in MSTO-211H cells were localized in the perinuclear space NP-C in combination with piroxicam induced apoptosis using a final cisplatin concentration 1.75 fold less than free drug. Delivered cisplatin cooperated with piroxicam in modulating cell cycle regulators as caspase-3, p53 and p21. Cisplatin loaded PLGA nanoparticles plus piroxicam showed a good efficacy in exerting cytotoxic activity and inducing the same molecular apoptotic effects of the free drugs. Sustained cisplatin release allowed to use less amount of drug, decreasing toxic side effects. This novel approach could represent a new strategy for mesothelioma treatment.


Pharmacotherapies for diabetes management: an update for the practicing clinician.
Over the past several years, the pharmacologic options for the management of glycemic control have tremendously expanded. Whereas prior to the introduction of metformin therapy in 1995 the only alternatives were human insulin therapies and sulfonylurea drugs, we now have the option of using several different classes of oral antidiabetic drugs, injectable non-insulin therapies, and insulin analogs. In this article we present a functional classification of glycemic therapies available for the treatment of diabetes in an attempt to provide the clinician with a practical framework which optimize blood glucose management. Patients with diabetes, especially type 2 diabetes, are often managed with a wide variety of injectable and non-injectable options in the attempt to improve glycemic control and glycemic variability, minimize hypoglycemia (as well as weight gain) and prevent both micro- and macrovascular complications. We specifically focus on novel therapies and present macrovascular complications. We specifically focus on novel therapies and present information about their efficacy and safety, as well as potential contraindications. The last section of this paper will present some suggestions for how to manage glycemia in the in-patient setting, including the use of rapid and long-acting insulin preparations to cover fasting and nutritional needs, as well as the proper use of insulin scales.


Correct and incorrect nucleotide incorporation pathways in DNA polymerase beta.
Tracking the structural and energetic changes in the pathways of DNA replication and repair is central to the understanding of these important processes. Here we report favorable mechanisms of the polymerase-catalyzed phosphoryl transfer reactions corresponding to correct and incorrect nucleotide incorporations in the DNA by using a novel protocol involving energy minimizations, dynamics simulations, quasi-harmonic free energy calculations, and mixed quantum mechanics/molecular mechanics dynamics simulations. Though the pathway proposed may not be unique and invites variations, geometric and energetic arguments support the series of transient intermediates in the phosphoryl transfer pathways uncovered here for both the G:C and G:A systems involving a Grotthuss hopping mechanism of proton transfer between water molecules and the three conserved aspartate residues in pol beta's active-site. In the G:C system, the rate-limiting step is the initial proton hop with a free energy of activation of at least 17 kcal/mol, which corresponds closely to measured k(pol) values. Fidelity discrimination in pol beta can be explained by a significant loss of stability of the closed ternary complex of the enzyme in the G:A system and much higher activation energy of the initial step of nucleophilic attack, namely deprotonation of terminal DNA primer O3'H group. Thus, subtle differences in the enzyme active-site between matched and mismatched base pairs generate significant differences in catalytic performance.


Proteolytic activities of human ADAMTS-5: comparative studies with ADAMTS-4.
Aggrecanases have been characterized as proteinases that cleave the Glu373-Ala374 bond of the aggrecan core protein, and they are multidomain metalloproteinases belonging to the ADAMTS (adamalysin with thrombospondin type 1 motifs) family. The first aggrecanases discovered were ADAMTS-4 (aggrecanase 1) and ADAMTS-5 (aggrecanase 2). They contain a zinc catalytic domain followed by non-catalytic ancillary domains, including a disintegrin domain, a thrombospondin domain, a cysteine-rich domain, and a spacer domain. In the case of ADAMTS-5, a second thrombospondin domain follows the spacer domain. We previously reported that the non-catalytic domains of ADAMTS-4 influence both its extracellular matrix interaction and proteolytic abilities. Here we report the effects of these domains of ADAMTS-5 on the extracellular matrix interaction and proteolytic activities and compare them with those of ADAMTS-4. Although the spacer domain was critical for ADAMTS-4 localization in the matrix, the cysteine-rich domain influenced ADAMTS-5 localization. Similar to previous reports of other ADAMTS family members, very little proteolytic activity was detected with the ADAMTS-5 catalytic domain alone. The sequential inclusion of each carboxyl-terminal domain enhanced its activity against aggrecan, carboxymethylated transferrin, fibromodulin, decorin, biglycan, and fibronectin. Both ADAMTS-4 and -5 had a broad optimal activity at pH 7.0-9.5. Aggrecanolytic activities were sensitive to the NaCl concentration, but activities on non-aggrecan substrates, e.g. carboxymethylated transferrin, were not affected. Although ADAMTS-4 and ADAMTS-5 had similar general proteolytic activities, the aggrecanase activity of ADAMTS-5 was at least 1,000-fold greater than that of ADAMTS-4 under physiological conditions. Our studies suggest that ADAMTS-5 is a major aggrecanase in cartilage metabolism and pathology.


On the "tertiary structure" of poly-carbenes; self-assembly of sp3-carbon-based polymers into liquid-crystalline aggregates.
The self-assembly of poly(ethylidene acetate) (st-PEA) into van der Waals-stabilized liquid-crystalline (LC) aggregates is reported. The LC behavior of these materials is unexpected, and unusual for flexible sp(3)-carbon backbone polymers. Although the dense packing of polar ester functionalities along the carbon backbone of st-PEA could perhaps be expected to lead directly to rigid-rod behavior, molecular modeling reveals that individual st-PEA chains are actually highly flexible and should not reveal rigid-rod induced LC behavior. Nonetheless, st-PEA clearly reveals LC behavior, both in solution and in the melt over a broad elevated temperature range. A combined set of experimental measurements, supported by MM/MD studies, suggests that the observed LC behavior is due to self-aggregation of st-PEA into higher-order aggregates. According to MM/MD modeling st-PEA single helices adopt a flexible helical structure with a preferred trans-gauche syn-syn-anti-anti orientation. Unexpectedly, similar modeling experiments suggest that three of these helices can self-assemble into triple-helical aggregates. Higher-order assemblies were not observed in the MM/MD simulations, suggesting that the triple helix is the most stable aggregate configuration. DLS data confirmed the aggregation of st-PEA into higher-order structures, and suggest the formation of rod-like particles. The dimensions derived from these light-scattering experiments correspond with st-PEA triple-helix formation. Langmuir-Blodgett surface pressure-area isotherms also point to the formation of rod-like st-PEA aggregates with similar dimensions as st-PEA triple helixes. Upon increasing the st-PEA concentration, the viscosity of the polymer solution increases strongly, and at concentrations above 20 wt % st-PEA forms an organogel. STM on this gel reveals the formation of helical aggregates on the graphite surface-solution interface with shapes and dimensions matching st-PEA triple helices, in good agreement with the structures proposed by molecular modeling. X-ray diffraction, WAXS, SAXS and solid state NMR spectroscopy studies suggest that st-PEA triple helices are also present in the solid state, up to temperatures well above the melting point of st-PEA. Formation of higher-order aggregates explains the observed LC behavior of st-PEA, emphasizing the importance of the "tertiary structure" of synthetic polymers on their material properties.


Reduced levels of DNA polymerase delta induce chromosome fragile site instability in yeast.
Specific regions of genomes (fragile sites) are hot spots for the chromosome rearrangements that are associated with many types of cancer cells. Understanding the molecular mechanisms regulating the stability of chromosome fragile sites, therefore, has important implications in cancer biology. We previously identified two chromosome fragile sites in Saccharomyces cerevisiae that were induced in response to the reduced expression of Pol1p, the catalytic subunit of DNA polymerase alpha. In the study presented here, we show that reduced levels of Pol3p, the catalytic subunit of DNA polymerase delta, induce instability at these same sites and lead to the generation of a variety of chromosomal aberrations. These findings demonstrate that a change in the stoichiometry of replicative DNA polymerases results in recombinogenic DNA lesions, presumably double-strand DNA breaks.


Exercise-based cardiac rehabilitation for adults after heart valve surgery.
The impact of exercise-based cardiac rehabilitation (CR) following heart valve surgery is uncertain. We conducted an update of this systematic review and a meta-analysis to assess randomised controlled trial evidence for the use of exercise-based CR following heart valve surgery. To assess the benefits and harms of exercise-based CR compared with no exercise training in adults following heart valve surgery or repair, including both percutaneous and surgical procedures. We considered CR programmes consisting of exercise training with or without another intervention (such as an intervention with a psycho-educational component). We searched the Cochrane Central Register of Clinical Trials (CENTRAL), in the Cochrane Library; MEDLINE (Ovid); Embase (Ovid); the Cumulative Index to Nursing and Allied Health Literature (CINAHL; EBSCO); PsycINFO (Ovid); Latin American Caribbean Health Sciences Literature (LILACS; Bireme); and Conference Proceedings Citation Index-Science (CPCI-S) on the Web of Science (Clarivate Analytics) on 10 January 2020. We searched for ongoing trials from ClinicalTrials.gov, Clinical-trials.com, and the World Health Organization International Clinical Trials Registry Platform on 15 May 2020. We included randomised controlled trials that compared exercise-based CR interventions with no exercise training. Trial participants comprised adults aged 18 years or older who had undergone heart valve surgery for heart valve disease (from any cause) and had received heart valve replacement or heart valve repair. Both percutaneous and surgical procedures were included. Two review authors independently extracted data. We assessed the risk of systematic errors ('bias') by evaluating risk domains using the 'Risk of bias' (RoB2) tool. We assessed clinical and statistical heterogeneity. We performed meta-analyses using both fixed-effect and random-effects models. We used the GRADE approach to assess the quality of evidence for primary outcomes (all-cause mortality, all-cause hospitalisation, and health-related quality of life). We included six trials with a total of 364 participants who have had open or percutaneous heart valve surgery. For this updated review, we identified four additional trials (216 participants). One trial had an overall low risk of bias, and we classified the remaining five trials as having some concerns. Follow-up ranged across included trials from 3 to 24 months. Based on data at longest follow-up, a total of nine participants died: 4 CR versus 5 control (relative risk (RR) 0.83, 95% confidence interval (CI) 0.26 to 2.68; 2 trials, 131 participants; GRADE quality of evidence very low). No trials reported on cardiovascular mortality. One trial reported one cardiac-related hospitalisation in the CR group and none in the control group (RR 2.72, 95% CI 0.11 to 65.56; 1 trial, 122 participants; GRADE quality of evidence very low). We are uncertain about health-related quality of life at completion of the intervention in CR compared to control (Short Form (SF)-12/36 mental component: mean difference (MD) 1.28, 95% CI -1.60 to 4.16; 2 trials, 150 participants; GRADE quality of evidence very low; and SF-12/36 physical component: MD 2.99, 95% CI -5.24 to 11.21; 2 trials, 150 participants; GRADE quality of evidence very low), or at longest follow-up (SF-12/36 mental component: MD -1.45, 95% CI -4.70 to 1.80; 2 trials, 139 participants; GRADE quality of evidence very low; and SF-12/36 physical component: MD -0.87, 95% CI -3.57 to 1.83; 2 trials, 139 participants; GRADE quality of evidence very low). AUTHORS' CONCLUSIONS: Due to lack of evidence and the very low quality of available evidence, this updated review is uncertain about the impact of exercise-CR in this population in terms of mortality, hospitalisation, and health-related quality of life. High-quality (low risk of bias) evidence on the impact of CR is needed to inform clinical guidelines and routine practice.


The relationship between measures of impulsivity and alcohol misuse: an integrative structural equation modeling approach.
Higher levels of impulsivity have been implicated in the development of alcohol use disorders. Recent findings suggest that impulsivity is not a unitary construct, highlighted by the diverse ways in which the various measures of impulsivity relate to alcohol use outcomes. This study simultaneously tested the following dimensions of impulsivity as determinants of alcohol use and alcohol problems: risky decision making, self-reported risk-attitudes, response inhibition, and impulsive decision making. Participants were a community sample of nontreatment seeking problem drinkers (n = 158). Structural equation modeling (SEM) analyses employed behavioral measures of impulsive decision making (delay discounting task [DDT]), response inhibition (stop signal task [SST]), and risky decision making (Balloon Analogue Risk Task [BART]), and a self-report measure of risk-attitudes (domain-specific risk-attitude scale [DOSPERT]), as predictors of alcohol use and of alcohol-related problems in this sample. The model fits well, accounting for 38% of the variance in alcohol problems, and identified 2 impulsivity dimensions that significantly loaded onto alcohol outcomes: (i) impulsive decision making, indexed by the DDT; and (ii) risky decision making, measured by the BART. The impulsive decision-making dimension of impulsivity, indexed by the DDT, was the strongest predictor of alcohol use and alcohol pathology in this sample of problem drinkers. Unexpectedly, a negative relationship was found between risky decision making and alcohol problems. The results highlight the importance of considering the distinct facets of impulsivity to elucidate their individual and combined effects on alcohol use initiation, escalation, and dependence.


The role of prior lexical knowledge in children's and adults' incidental word learning from illustrated stories.
Children and adults benefit from a new word's phonological neighbors during explicit vocabulary instruction, suggesting that related prior knowledge can support new learning. This study examined the influence of lexical neighborhood structure during incidental word learning-limiting opportunities for strategically engaging prior knowledge-and tested the hypothesis that prior knowledge would provide additional support during subsequent consolidation. Children aged 8-10 years (Experiment 1) and adults (Experiment 2) were presented with 15 pseudowords embedded in a spoken story with illustrations, and were then tested on their recognition and recall of the new word-forms immediately, the next day, and one week later. The pseudowords had either no, one, or many English phonological neighbors, varying the potential connections to existing knowledge. After encountering the pseudowords in this incidental training paradigm, neither children nor adults benefited from phonological neighbors in recall, and children were better at recognizing items without neighbors. The neighbor influence did not change with opportunities for consolidation in either experiment, nor did it relate to learners' existing vocabulary ability. Exploratory analyses revealed that children experienced bigger benefits from offline consolidation overall, with adults outperforming children only for many-neighbor items one week after exposure. We discuss how the neighbor benefit in word learning may be constrained by learning context, and how the enhanced benefits of offline consolidation in childhood extend to vocabulary learning in more naturalistic contexts. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Human Recombinant VEGFR2D4 Biochemical Characterization to Investigate Novel Anti-VEGFR2D4 Antibodies for Allosteric Targeting of VEGFR2.
VEGF-A/VEGFR2 complex is the major signaling pathway involved in angiogenesis and the inhibition of this axis retards tumor growth and inflammatory disorders progression, reducing vessel sprouting. Signaling by VEGFR2 requires receptor dimerization and a well-defined orientation of monomers in the active dimer. The extracellular portion of receptor is composed of seven Ig-like domains, of which D2-3 are the ligand binding domains, while D4 and D7, establishing homotypic contacts, allosterically regulate receptor activity. The allosteric targeting of VEGFR2 represents a promising alternative to study neovascular disorders overcoming drawbacks related to competition with VEGF. In this work, we expressed in bacterial host domain 4 of VEGFR2 (VEGFR2D4). After protein refolding, we characterized the purified domain and administered it in mice for monoclonal antibodies production. One of them, mAbD4, was tested in ELISA assays, showing a nanomolar affinity for VEGFR2D4. Finally, the methodology here described could contribute to the development of antibodies which can allosterically bind VEGFR2 and therefore to be used for imaging purposes or to modulate receptor signaling.


The Capabilities and Limitations of Clinical Magnetic Resonance Imaging for Detecting Kidney Stones: A Retrospective Study.
The purpose of this work was to investigate the performance of currently available magnetic resonance imaging (MRI) for detecting kidney stones, compared to computed tomography (CT) results, and to determine the characteristics of successfully detected stones. Patients who had undergone both abdominal/pelvic CT and MRI exams within 30 days were studied. The images were reviewed by two expert radiologists blinded to the patients' respective radiological diagnoses. The study consisted of four steps: (1) reviewing the MRI images and determining whether any kidney stone(s) are identified; (2) reviewing the corresponding CT images and confirming whether kidney stones are identified; (3) reviewing the MRI images a second time, armed with the information from the corresponding CT, noting whether any kidney stones are positively identified that were previously missed; (4) for all stones MRI-confirmed on previous steps, the radiologist experts being asked to answer whether in retrospect, with knowledge of size and location on corresponding CT, these stones would be affirmed as confidently identified on MRI or not. In this best-case scenario involving knowledge of stones and their locations on concurrent CT, radiologist experts detected 19% of kidney stones on MRI, with stone size being a major factor for stone identification.


Autosomal dominant cerulean cataract is associated with a chain termination mutation in the human beta-crystallin gene CRYBB2.
Congenital cataracts are a common major abnormality of the eye that frequently cause blindness in infants. At least a third of all cases are familial; autosomal dominant congenital cataract (ADCC) appears to be the most common familial form in the Western world. Cerulean cataracts have peripheral bluish and white opacifications in concentric layers with occasional central lesions arranged radially. Although the opacities may be observed during fetal development and childhood, usually visual acuity is only mildly reduced until adulthood, when lens extraction is generally necessary. We have been studying a family (ADCC-1) with cerulean blue ADCC, in which the affected daughter of a first cousin mating was presumed to be homozygous for the cataract gene. Recently, we mapped an ADCC gene in this family to a region of chromosome 22 containing three beta-crystallin genes. Here we report that a chain-termination mutation in CRYBB2 is associated with ADCC in this family.


Resistance to different classes of drugs is associated with impaired apoptosis in childhood acute lymphoblastic leukemia.
Resistance of leukemic cells to chemotherapeutic agents is associated with an unfavorable outcome in pediatric acute lymphoblastic leukemia (ALL). To investigate the underlying mechanisms of cellular drug resistance, the activation of various apoptotic parameters in leukemic cells from 50 children with ALL was studied after in vitro exposure with 4 important drugs in ALL therapy (prednisolone, vincristine, l-asparaginase, and daunorubicin). Exposure to each drug resulted in early induction of phosphatidylserine (PS) externalization and mitochondrial transmembrane (Deltapsim) depolarization followed by caspase-3 activation and poly(ADP-ribose) polymerase (PARP) inactivation in the majority of patients. For all 4 drugs, a significant inverse correlation was found between cellular drug resistance and (1) the percentage of cells with PS externalization (<.001 < P <.008) and (2) the percentage of cells with Deltapsim depolarization (.002 < P <.02). However, the percentage of cells with caspase-3 activation and the percentage of cells with PARP inactivation showed a significant inverse correlation with cellular resistance for prednisolone (P =.001; P =.001) and l-asparaginase (P =.01; P =.001) only. This suggests that caspase-3 activation and PARP inactivation are not essential for vincristine- and daunorubicin-induced apoptosis. In conclusion, resistance to 4 unrelated drugs is associated with defect(s) upstream or at the level of PS externalization and Deltapsim depolarization. This leads to decreased activation of apoptotic parameters in resistant cases of pediatric ALL.


Stability of underdominant genetic polymorphisms in population networks.
Heterozygote disadvantage is potentially a potent driver of population genetic divergence. Also referred to as underdominance, this phenomena describes a situation where a genetic heterozygote has a lower overall fitness than either homozygote. Attention so far has mostly been given to underdominance within a single population and the maintenance of genetic differences between two populations exchanging migrants. Here we explore the dynamics of an underdominant system in a network of multiple discrete, yet interconnected, populations. Stability of genetic differences in response to increases in migration in various topological networks is assessed. The network topology can have a dominant and occasionally non-intuitive influence on the genetic stability of the system.


Insights and limits of translational research in critical care medicine.
Experimental research has always been the cornerstone of pathophysiological and therapeutic advances in critical care medicine, where clinical observations and basic research mutually fed each other in a so-called translational approach. The objective of this review is to address the different aspects of translational research in the field of critical care medicine. We herein highlighted some demonstrative examples including the animal-to-human approach to study host-pathogen interactions, the human-to-animal approach for sepsis-induced immunosuppression, the still restrictive human approach to study critical illness-related neuromyopathy, and the technological developments to assess the microcirculatory changes in critically ill patients. These examples not only emphasize how translational research resulted in major improvements in the comprehension of the pathophysiology of severe clinical conditions and offered promising perspectives in critical care medicine but also point out the obstacles to translate such achievements into clinical practice.


Combination therapies with thiazolidinediones are associated with a lower risk of acute exacerbations in new-onset COPD patients with advanced diabetic mellitus: a cohort-based case-control study.
The effects of oral antihyperglycaemic drugs (OADs) for type 2 diabetes mellitus (T2DM) on the outcomes of co-existing chronic obstructive pulmonary disease (COPD) patients are not well studied. We examined the association of combinational OADs and the risk of acute exacerbations of COPD (AECOPD) in T2DM patients with co-existing COPD. A cohort-based case-control study was conducted using data from the National Health Insurance Research Database of Taiwan. Among new-onset COPD-T2DM patients, 65,370 were prescribed metformin and 2nd-line OADs before the date of COPD onset. Each AECOPD case was matched to 4 randomly selected controls according to the propensity score estimated by the patient's baseline characteristics. Conditional logistic regression analysis was performed to estimate the association between AECOPD risk and OAD use. Among COPD-T2DM patients, 3355 AECOPD cases and 13,420 matched controls were selected. Of the patients treated with a double combination of oral OADs (n = 12,916), those treated with sulfonylurea (SU) and thiazolidinediones (TZD) had a lower AECOPD risk than the patients who received metformin (MET) and SU, with an adjusted odds ratio (OR) of 0.69 (95% confidence interval [CI] 0.51-0.94, P = 0.02). Of the patients with a triple combination of oral OADs (n = 3859), we found that those treated with MET, SU and TZD had a lower risk of AECOPD (adjusted OR 0.81 (0.68-0.96, P = 0.01) than a combination of MET, SU and α-glucosidase inhibitors (AGIs) regardless of the level of COPD complexity. Combination therapies with TZD were associated with a reduced risk of AECOPD in advanced T2DM patients with co-existing COPD.


Isolation of Escherichia coli RNA polymerase binding sites on T5 and T7 DNA: further evidence for sigma-dependent recognition of A-T-rich DNA sequences.
Previous isolation and analysis of E. coli RNA polymerase (EC 18.104.22.168) binding sites on lambda DNA had demonstrated the existence of a sigma-dependent process of recognition of A-T-rich DNA sequences. We have now extended this finding to T5 and T7 DNA and häve provided evidence for the double-strandedness of the isolated binding sites. The possible equation of these sites to the genetically defined promoters is discussed.


H3Africa multi-centre study of the prevalence and environmental and genetic determinants of type 2 diabetes in sub-Saharan Africa: study protocol.
The burden and aetiology of type 2 diabetes (T2D) and its microvascular complications may be influenced by varying behavioural and lifestyle environments as well as by genetic susceptibility. These aspects of the epidemiology of T2D have not been reliably clarified in sub-Saharan Africa (SSA), highlighting the need for context-specific epidemiological studies with the statistical resolution to inform potential preventative and therapeutic strategies. Therefore, as part of the Human Heredity and Health in Africa (H3Africa) initiative, we designed a multi-site study comprising case collections and population-based surveys at 11 sites in eight countries across SSA. The goal is to recruit up to 6000 T2D participants and 6000 control participants. We will collect questionnaire data, biophysical measurements and biological samples for chronic disease traits, risk factors and genetic data on all study participants. Through integrating epidemiological and genomic techniques, the study provides a framework for assessing the burden, spectrum and environmental and genetic risk factors for T2D and its complications across SSA. With established mechanisms for fieldwork, data and sample collection and management, data-sharing and consent for re-approaching participants, the study will be a resource for future research studies, including longitudinal studies, prospective case ascertainment of incident disease and interventional studies.


The effects of Cetraria islandica and Pseudevernia furfuracea extracts in normal and diabetic rats.
Lichens are symbiotic organisms composed of a fungus joined to a photosynthesizing partner that can be either an alga or a cyanobacterium. They can be used as a novel bioresource for natural antioxidants. However, there is also a need for further studies to validate the lichens used in medicinal remedies. This study covers a previously unrecognized effects of Cetraria islandica (CIAE) and Pseudevernia furfuracea (PFAE) in streptozotocin (STZ)-induced diabetes. In experimental design, control or diabetic rats were either untreated or treated with aqueous lichen extracts (250-500 mg/kg/day) for 2 weeks starting at 72 h after STZ injection. On day 14, animals were anesthetized, metabolic and biochemical parameters were appreciated between control and treatment groups. The histopathology of kidney was examined using four different staining methods: hematoxylin-eosin (H&E), periodic acid-Schiff (PAS), Masson trichrome and Congo red. Our experimental data showed that increasing doses of CIAE and PFAE did not have any detrimental effects on the studied parameters and the malondialdehyde level of kidney. CIAE extract showed prominent results compared to doses of PFAE extract for antioxidant capacity. However, the protective effect of CIAE extract was inadequate on diabetes-induced disorders and kidney damages. Moreover, animals subjected to diabetes mellitus (DM) therapy did not benefit unfortunately from the usage of increasing lichen doses due to their unchanged antioxidant activity to tissue. The results obtained in present study suggested that CIAE and PFAE are safe but the power of these is limited because of the intensive oxidative stress in kidney of type 1 diabetic rats. It is also implied that CIAE extract is especially suitable for different administration routes in DM.


Sirtuins: novel targets for metabolic disease in drug development.
Calorie restriction extends lifespan and produces a metabolic profile desirable for treating diseases such as type 2 diabetes. SIRT1, an NAD(+)-dependent deacetylase, is a principal modulator of pathways downstream of calorie restriction that produces beneficial effects on glucose homeostasis and insulin sensitivity. Activation of SIRT1 leads to enhanced activity of multiple proteins, including peroxisome proliferator-activated receptor coactivator-1alpha (PGC-1alpha) and FOXO which helps to mediate some of the in vitro and in vivo effects of sirtuins. Resveratrol, a polyphenolic SIRT1 activator, mimics the effects of calorie restriction in lower organisms and in mice fed a high-fat diet ameliorates insulin resistance. In this review, we summarize recent research advances in unveiling the molecular mechanisms that underpin sirtuin as therapeutic candidates and discuss the possibility of using resveratrol as potential drug for treatment of diabetes.


Prognostic and biologic significance of chromosomal imbalances assessed by comparative genomic hybridization in multiple myeloma.
Cytogenetic abnormalities, evaluated either by karyotype or by fluorescence in situ hybridization (FISH), are considered the most important prognostic factor in multiple myeloma (MM). However, there is no information about the prognostic impact of genomic changes detected by comparative genomic hybridization (CGH). We have analyzed the frequency and prognostic impact of genetic changes as detected by CGH and evaluated the relationship between these chromosomal imbalances and IGH translocation, analyzed by FISH, in 74 patients with newly diagnosed MM. Genomic changes were identified in 51 (69%) of the 74 MM patients. The most recurrent abnormalities among the cases with genomic changes were gains on chromosome regions 1q (45%), 5q (24%), 9q (24%), 11q (22%), 15q (22%), 3q (16%), and 7q (14%), while losses mainly involved chromosomes 13 (39%), 16q (18%), 6q (10%), and 8p (10%). Remarkably, the 6 patients with gains on 11q had IGH translocations. Multivariate analysis selected chromosomal losses, 11q gains, age, and type of treatment (conventional chemotherapy vs autologous transplantation) as independent parameters for predicting survival. Genomic losses retained the prognostic value irrespective of treatment approach. According to these results, losses of chromosomal material evaluated by CGH represent a powerful prognostic factor in MM patients.


Bicycle crashes in different riding environments in the Australian capital territory.
Cyclists are increasingly overrepresented in traffic crash casualties in Australia. There is evidence that better cycling infrastructure increases participation, but whether it reduces the numbers of injured cyclists is less clear. This study examined injury outcomes of crashes in different cycling environments. Adult cyclists injured on- and off-road were recruited from emergency departments from November 2009 to May 2010 in the Australian Capital Territory. Eligible participants (n = 313/372, 84.1%) were interviewed and their injury self-reports were corroborated with medical records where available. Participants who had crashed in transport-related areas (n = 202, 64.5%) are the focus of this article. Participants had crashed in traffic (39.1%), in cycle lanes (7.9%), on shared paths (36.1%), and on footpaths (16.8%). Based on average weekly traffic counts, the crash involvement rate per 1000 cyclists was 11.8 on shared paths compared to 5.8 on cycle lanes. Over half of the participants (52.0%) were injured in single-vehicle bicycle crashes. The remainder involved other road users, including motor vehicles (20.8%), other bicycles (18.8%), pedestrians (6.4%), and animals (2.0%). Pedestrians were involved in 16.4 percent of crashes on shared paths. Minor injuries (Abbreviated Injury Scale [AIS] 1) were sustained by 58.4 percent of cyclists, moderately severe injuries (AIS 2) were sustained by 36.1 percent of cyclists, and 5.4 percent of cyclists were seriously injured (AIS 3+). The average treatment required was 1.8 days with 7.5 days off work and cost to the cyclist of $869 excluding medical treatment. Cyclists who crashed on shared paths or in traffic had higher injury severity scores (ISS; 4.4, 4.0) compared to those in cycle lanes or on footpaths (3.3, 3.4) and required more treatment days (2.8, 1.7 versus 0.0, 0.2). Fewer cyclists were injured in on-road cycle lanes than in other cycling environments, and a high proportion of injuries were incurred on shared paths. This study highlights an urgent need to determine appropriate criteria and management strategies for paths classified as suitable for shared or segregated usage. Supplemental materials are available for this article. Go to the publisher's online edition of Traffic Injury Prevention to view the supplemental files.


Extension as a Backbone Support Organization for Physical Activity Promotion: A Collective Impact Case Study From Rural Kentucky.
Rural counties in the United States face daunting structural issues that reduce their populations' physical activity levels, including geographic isolation as well as deficits in infrastructure, public transportation, health care providers, and funding. Funding from the Centers for Disease Control and Prevention provided an opportunity to assess how Extension enhanced the collective impact of systems-level physical activity promotion programming through a multisectoral coalition in Clinton County, Kentucky. The Extension-led coalition accomplished the 6 essential functions of a backbone support organization by identifying obesity as a critical local issue (function 1: providing overall strategic direction), developing a multisectoral coalition (function 2: facilitating dialog between partners), compiling data on the county's physical activity infrastructure (function 3: managing data collection and analysis), creating communication channels (function 4: handling communication), organizing community awareness events (function 5: coordinating community outreach), and securing additional grants (function 6: mobilizing funding). The average rating of Extension's leadership across multiple dimensions by 3 coalition members in a postproject survey was "excellent" on a 5-point Likert scale. Extension is well positioned through their mission, broad community engagement, data collection, needs assessment, community and academic relationships, and embeddedness in local communities to serve as the backbone support organizations for rural physical activity promotion coalitions.


Incidence, risk factors and impact of seasonal influenza in pregnancy: A national cohort study.
Pregnant women are particularly vulnerable to severe infection from influenza resulting in poor neonatal outcomes. The majority of evidence relates to pandemic 2009 A/H1N1 influenza. The objective of this study was to describe the characteristics and outcomes of pregnant women hospitalised with seasonal influenza. This national, prospective, observational cohort study used the UK Obstetric Surveillance System (UKOSS) to identify all pregnant women admitted to hospital between 01/11/2016 and 31/10/2018 with laboratory confirmed influenza at any gestation and up to two days after giving birth. These were compared to women admitted to give birth that did not have influenza. Baseline characteristics, immunization status, maternal and perinatal outcomes were compared. There were 405 women admitted to hospital with laboratory confirmed influenza in pregnancy: 2.7 per 10,000 maternities. Compared to 694 comparison women, women with influenza were less likely to be professionally employed (aOR 0.59, 95%CI 0.39-0.89) or immunised in the relevant season (aOR 0·59, 0·39-0·89) and more likely to have asthma (aOR 2.42, 1.30-4.49) or have had a previous pregnancy complication (aOR 2·47, 1·33-4·61). They were more likely to be admitted to intensive care (aOR 21.3, 2.78-163.1) and to have a cesarean birth (aOR 1·42, 1·02-1.98). Their babies were more likely to be admitted to neonatal intensive care (aOR 1.86, 1·01-3·42). Immunization reduces the risk of hospitalisation with influenza in pregnancy which is associated with increased risk of morbidity for both the mother and baby. There is a continued need to increase awareness of safety and effectiveness of immunization in pregnancy and provision within antenatal care settings, especially for high-risk groups.


Overexpression of guanylate cyclase activating protein 2 in rod photoreceptors in vivo leads to morphological changes at the synaptic ribbon.
Guanylate cyclase activating proteins are EF-hand containing proteins that confer calcium sensitivity to retinal guanylate cyclase at the outer segment discs of photoreceptor cells. By making the rate of cGMP synthesis dependent on the free intracellular calcium levels set by illumination, GCAPs play a fundamental role in the recovery of the light response and light adaptation. The main isoforms GCAP1 and GCAP2 also localize to the synaptic terminal, where their function is not known. Based on the reported interaction of GCAP2 with Ribeye, the major component of synaptic ribbons, it was proposed that GCAP2 could mediate the synaptic ribbon dynamic changes that happen in response to light. We here present a thorough ultrastructural analysis of rod synaptic terminals in loss-of-function (GCAP1/GCAP2 double knockout) and gain-of-function (transgenic overexpression) mouse models of GCAP2. Rod synaptic ribbons in GCAPs-/- mice did not differ from wildtype ribbons when mice were raised in constant darkness, indicating that GCAPs are not required for ribbon early assembly or maturation. Transgenic overexpression of GCAP2 in rods led to a shortening of synaptic ribbons, and to a higher than normal percentage of club-shaped and spherical ribbon morphologies. Restoration of GCAP2 expression in the GCAPs-/- background (GCAP2 expression in the absence of endogenous GCAP1) had the striking result of shortening ribbon length to a much higher degree than overexpression of GCAP2 in the wildtype background, as well as reducing the thickness of the outer plexiform layer without affecting the number of rod photoreceptor cells. These results indicate that preservation of the GCAP1 to GCAP2 relative levels is relevant for maintaining the integrity of the synaptic terminal. Our demonstration of GCAP2 immunolocalization at synaptic ribbons at the ultrastructural level would support a role of GCAPs at mediating the effect of light on morphological remodeling changes of synaptic ribbons.