Comparative experimental transmission of cardiomyopathy syndrome (CMS) in Atlantic salmon Salmo salar.
Cardiomyopathy syndrome (CMS) has been recorded in wild and farmed Atlantic salmon Salmo salar. Characteristic heart lesions primarily involving the myocardium are reported in natural outbreaks with associated mortality. To date, no experimental trials have reproduced these lesions in the laboratory. The present study reports on the first successful experimental transmission of CMS in Atlantic salmon in Scotland, with full development of the histological lesions that are described for the syndrome. Tissue homogenates of CMS-infected fish indicative of mild and severe lesions from Scottish and Norwegian natural outbreaks, respectively, were injected into naïve fish, and both induced heart lesions consistent with CMS. Lesion development was earlier and progression faster in the fish group receiving the Norwegian homogenate, but equivalent in both groups by the end time point of the experiment. The study demonstrated that the reported condition for both countries is identical, as evaluated through light microscopy, and that tissue homogenates from either mild or severely affected fish contain the transmissible agent.


Genomic effects on advertisement call structure in diploid and triploid hybrid waterfrogs (Anura, Pelophylax esculentus).
In anurans, differences in male mating calls have intensively been studied with respect to taxonomic classification, phylogeographic comparisons among different populations and sexual selection. Although overall successful, there is often much unexplained variation in these studies. Potential causes for such variation include differences among genotypes and breeding systems, as well as differences between populations. We investigated how these three factors affect call properties in male water frogs of Pelophylax lessonae (genotype LL), P. ridibundus (RR) and their interspecific hybrid P. esculentus which comes in diploid (LR) and triploid types (LLR, LRR). We investigated five call parameters that all showed a genomic dosage effect, i.e. they either decreased or increased with the L/R ratio in the order LL-LLR-LR-LRR-RR. Not all parameters differentiated equally well between the five genotypes, but combined they provided a good separation. Two of the five call parameters were also affected by the breeding system. Calls of diploid LR males varied, depending on whether these males mated with one or both of the parental species (diploid systems) or triploid hybrids (mixed ploidy systems). With the exception of the northernmost mixed-ploidy population, call differences were not related to the geographic location of the population and they were not correlated with genetic distances in the R and L genomes. We found an influence of all three tested factors on call parameters, with the effect size decreasing from genotype through breeding system to geographic location of the population. Overall, results were in line with predictions from a dosage effect in L/R ratios, but in three call parameters all three hybrid types were more similar to one or the other parental species. Also calls of diploid hybrids varied between breeding systems in agreement with the sexual host required for successful reproduction. The lack of hybrid call differences in a mixed-ploidy population at the northern edge of the water frog distribution is likely to be associated with genetic particularities, including a) low genetic variability and/or b) a local loss of genes coding for genotype-dependent call differentiation under conditions where female discrimination between diploid and triploid males is not beneficial.


LeuT-desipramine structure reveals how antidepressants block neurotransmitter reuptake.
Tricyclic antidepressants exert their pharmacological effect-inhibiting the reuptake of serotonin, norepinephrine, and dopamine-by directly blocking neurotransmitter transporters (SERT, NET, and DAT, respectively) in the presynaptic membrane. The drug-binding site and the mechanism of this inhibition are poorly understood. We determined the crystal structure at 2.9 angstroms of the bacterial leucine transporter (LeuT), a homolog of SERT, NET, and DAT, in complex with leucine and the antidepressant desipramine. Desipramine binds at the inner end of the extracellular cavity of the transporter and is held in place by a hairpin loop and by a salt bridge. This binding site is separated from the leucine-binding site by the extracellular gate of the transporter. By directly locking the gate, desipramine prevents conformational changes and blocks substrate transport. Mutagenesis experiments on human SERT and DAT indicate that both the desipramine-binding site and its inhibition mechanism are probably conserved in the human neurotransmitter transporters.


Propofol inhibits the expression of Abelson nonreceptor tyrosine kinase without affecting learning or memory function in neonatal rats.
Propofol is one of the most commonly used intravenous drugs to induce and maintain general anesthesia. In vivo and in vitro studies have shown that propofol can affect neuronal growth, leading to apoptosis and impairing cognitive function. The Abelson nonreceptor tyrosine kinase (c-Abl) is associated with both neuritic plaques and neurofibrillary tangles in the brains of patients with Alzheimer's disease and other neurodegenerative diseases. This study aimed to explore the effect of propofol on apoptosis and neurocognition through its regulation of c-Abl expression in vivo and in vitro. In this study, primary hippocampal neurons were cultured and exposed to propofol at different concentrations. Protein expression was measured by Western blotting and coimmunoprecipitation. The c-Abl transcription level was verified by fluorescence quantitative PCR. Reactive oxygen species (ROS) levels were detected by flow cytometry. In addition, an animal experiment was conducted to assess neuronal apoptosis by immunofluorescence staining for caspase-3 and to evaluate behavioral changes by the Morris water maze (MWM) test. The in vitro experiment showed that propofol significantly decreased c-Abl expression and ROS levels. In addition, propofol has no cytotoxic effect and does not affect cell activity. Moreover, in the animal experiment, intraperitoneal injection of 50 mg/kg propofol for 5 days obviously decreased the expression of c-Abl in the neonatal rat brain (p < .05) but did not significantly increase the number of caspase-3-positive cells. Propofol treatment did not significantly reduce the number of platform crossings (p > .05) or prolong the escape latency of neonatal rats (p > .05) in the MWM test. The present data suggest that reduced expression of this nonreceptor tyrosine kinase through consecutive daily administration of propofol did not impair learning or memory function in neonatal rats.


Lifetime prevalence, age of risk, and genetic relationships of comorbid psychiatric disorders in Tourette syndrome.
Tourette syndrome (TS) is characterized by high rates of psychiatric comorbidity; however, few studies have fully characterized these comorbidities. Furthermore, most studies have included relatively few participants (<200), and none has examined the ages of highest risk for each TS-associated comorbidity or their etiologic relationship to TS. To characterize the lifetime prevalence, clinical associations, ages of highest risk, and etiology of psychiatric comorbidity among individuals with TS. Cross-sectional structured diagnostic interviews conducted between April 1, 1992, and December 31, 2008, of participants with TS (n = 1374) and TS-unaffected family members (n = 1142). Lifetime prevalence of comorbid DSM-IV-TR disorders, their heritabilities, ages of maximal risk, and associations with symptom severity, age at onset, and parental psychiatric history. The lifetime prevalence of any psychiatric comorbidity among individuals with TS was 85.7%; 57.7% of the population had 2 or more psychiatric disorders. The mean (SD) number of lifetime comorbid diagnoses was 2.1 (1.6); the mean number was 0.9 (1.3) when obsessive-compulsive disorder (OCD) and attention-deficit/hyperactivity disorder (ADHD) were excluded, and 72.1% of the individuals met the criteria for OCD or ADHD. Other disorders, including mood, anxiety, and disruptive behavior, each occurred in approximately 30% of the participants. The age of greatest risk for the onset of most comorbid psychiatric disorders was between 4 and 10 years, with the exception of eating and substance use disorders, which began in adolescence (interquartile range, 15-19 years for both). Tourette syndrome was associated with increased risk of anxiety (odds ratio [OR], 1.4; 95% CI, 1.0-1.9; P = .04) and decreased risk of substance use disorders (OR, 0.6; 95% CI, 0.3-0.9; P = .02) independent from comorbid OCD and ADHD; however, high rates of mood disorders among participants with TS (29.8%) may be accounted for by comorbid OCD (OR, 3.7; 95% CI, 2.9-4.8; P < .001). Parental history of ADHD was associated with a higher burden of non-OCD, non-ADHD comorbid psychiatric disorders (OR, 1.86; 95% CI, 1.32-2.61; P < .001). Genetic correlations between TS and mood (RhoG, 0.47), anxiety (RhoG, 0.35), and disruptive behavior disorders (RhoG, 0.48), may be accounted for by ADHD and, for mood disorders, by OCD. This study is, to our knowledge, the most comprehensive of its kind. It confirms the belief that psychiatric comorbidities are common among individuals with TS, demonstrates that most comorbidities begin early in life, and indicates that certain comorbidities may be mediated by the presence of comorbid OCD or ADHD. In addition, genetic analyses suggest that some comorbidities may be more biologically related to OCD and/or ADHD rather than to TS.


Management of male urinary incontinence.
The majority of male urinary incontinence seen is secondary to sphincter weakness following prostatic surgery. As there is a rising elderly population and increasing numbers of surgical interventions for prostate cancer, incidence of male incontinence is increasing. Hence, management of male incontinence has become a subject of increased interest for urologists. Various non-surgical and surgical approaches have been suggested for this devastating condition. Non-invasive therapies are suggested for early postoperative and mild incontinence. For surgical treatment the artificial urinary sphincter is still labeled the gold standard despite the introduction of several more minimally invasive treatments. However, as yet there is no consensus on the optimal timing and best modality for managing these men. Well designed, centrally funded clinical trials are required to establish which treatment modality to offer and when in the broad spectrum of male incontinence. This review focuses mainly on the management of post-prostatectomy incontinence since the management of other types varies little from the modalities of treatment in women.


Eczema in early childhood, sociodemographic factors and lifestyle habits are associated with food allergy: a nested case-control study.
Studies suggest an increase in food allergy prevalence over the last decade, but the contributing factors remain unknown. The aim of this study was to evaluate the association between the most common food allergies and atopic history, sociodemographic characteristics and lifestyle habits. We conducted a case-control study nested within the SPAACE study (Surveying Prevalence of Food Allergy in All Canadian Environments) – a cross-Canada, random telephone survey. Cases consisted of individuals with probable food allergy (self-report of convincing symptoms and/or physician diagnosis) to milk, egg, peanut, tree nut, shellfish, fish, wheat, soy, or sesame. Controls consisted of nonallergic individuals, matched for age. Cases and controls were queried on personal and family history of atopy, sociodemographic characteristics and lifestyle habits. Multivariate logistic regression was used to evaluate the association between atopy, sociodemographic characteristics and lifestyle habits with probable food allergy. Between September 2010 and September 2011, 480 cases and 4,950 controls completed the questionnaire. For all 9 allergens, factors associated with a higher risk of probable allergy were as follows: (1) personal history of eczema (in the first 2 years of life), asthma or hay fever (odds ratio, OR 2.3, 95% CI 1.6-3.5; OR 2.8, 95% CI 2.2-3.6, and OR 2.3, 95% CI 1.8-3.0, respectively), (2) maternal, paternal or sibling's food allergy (OR 3.7, 95% CI 2.5-5.6; OR 3.0, 95% CI 1.8-5.1, and OR 3.1, 95% CI 2.2-4.2), (3) high household income (top 20%; OR 1.5, 95% CI 1.2-2.0). Males and older individuals were less likely to have food allergy (OR 0.7, 95% CI 0.6-0.9, and OR 0.99, 95% CI 0.99-1.00). Eczema in the first 2 years of life was the strongest risk factor for egg, peanut, tree nut and fish allergy. This is the largest population-based nested case-control study exploring factors associated with food allergies. Our results reveal that, in addition to previously reported factors, eczema in the first 2 years of life is consistently associated with food allergies.


Therapeutic role of coenzyme Q(10) in Parkinson's disease.
Mitochondrial dysfunction has been well established to occur in Parkinson's disease (PD) and appears to play a role in the pathogenesis of the disorder. A key component of the mitochondrial electron transport chain (ETC) is coenzyme Q(10), which not only serves as the electron acceptor for complexes I and II of the ETC but is also an antioxidant. In addition to being crucial to the bioenergetics of the cell, mitochondria play a central role in apoptotic cell death through a number of mechanisms, and coenzyme Q(10) can affect certain of these processes. Levels of coenzyme Q(10) have been reported to be decreased in blood and platelet mitochondria from PD patients. A number of preclinical studies in in vitro and in vivo models of PD have demonstrated that coenzyme Q(10) can protect the nigrostriatal dopaminergic system. A phase II trial of coenzyme Q(10) in patients with early, untreated PD demonstrated a positive trend for coenzyme Q(10) to slow progressive disability that occurs in PD.


PTK2 rs7460 and rs7843014 polymorphisms and exceptional longevity: a functional replication study.
Focal adhesion is critical for cell survival. The focal adhesion kinase (FAK, or PTK2) is an important component of the human interactome and thus is a potential longevity-related protein. Here we studied the association between two PTK2 gene single-nucleotide polymorphisms (SNPs) (rs7843014, rs7460) and exceptional longevity (EL). In addition to gaining insight into their functionality by determining luciferase gene reporter activity, we studied the genotype/allele frequency of these two SNPs among three different cohorts: (1) Spanish centenarians (n=175, 100-111 years, 144 women) and healthy controls (n=355, 20-50 years, 284 women); (2) Italian centenarians (n=79, 100-104 years, 40 women) and controls (n=316, 29-50 years, 156 women); and (3) Japanese centenarians (n=742, 100-116 years, 623 women) and healthy controls (n=499, 23-59 years, 356 women). Both SNPs had functional significance, with the A allele up-regulating luciferase activity compared to the other allele (rs7460 T allele and rs7843014 C allele, respectively). The A allele of both SNPs was negatively associated with EL in the Spanish cohort (rs7460, odds ratio [OR] adjusted by sex=0.40, 95% confidence intervals [CI] 0.3, 0.6, p<0.001); rs7843014, OR=0.37, 95% CI 0.3, 0.5, p<0.001). The OR of being a centenarian if having the rs7460-TT genotype was 6.68 (95% CI 4.1, 10.8, p<0.001). The rs7843014 CC genotype was also positively associated with EL (OR=7.58, 95% CI 4.6, 12.3, p<0.001]. No association was, however, found for the Italian or Japanese cohorts. Thus, two genotypes of the FAK gene, rs7460 TT and rs7843014 CC, are possibly associated with lower gene expression and might favor the likelihood of reaching EL in the Spanish population. Further research is needed to unveil the mechanisms by which FAK expression could perhaps influence the rate of aging.


Corrector systems for cassegrain telescopes.
Most modern reflecting telescopes have relative apertures of about f/3 and f/8 for the primary and first secondary foci in accordance with the suggestions of Bowen. The angular field which can be used at the first secondary focus is limited by the size of available plates for large instruments but can approach +/-1 degrees for smaller systems. The factors influencing the choice of the field corrector system in the first secondary focus are discussed. It is an important point whether the Ritchey-Chrétien form of the mirrors is strictly maintained-giving an optimum field without the corrector-or whether the aspheric constants are allowed to vary as free parameters. The differences are small but significant. The performance of a number of secondary focus correctors consisting of one, two, and three elements is discussed, spot diagrams being given in each case. Systems with fixed Ritchey-Chrétien mirror constants are inferior to those with free mirror constants. Test methods for the manufacture of the mirrors of telescopes of this type are compared. A doublet type corrector is suitable for compensation testing of primary mirrors or for secondaries tested from the back, but the testing of the latter from the front is more difficult. Several possible techniques are discussed.


Intact flagellar motor of Borrelia burgdorferi revealed by cryo-electron tomography: evidence for stator ring curvature and rotor/C-ring assembly flexion.
The bacterial flagellar motor is a remarkable nanomachine that provides motility through flagellar rotation. Prior structural studies have revealed the stunning complexity of the purified rotor and C-ring assemblies from flagellar motors. In this study, we used high-throughput cryo-electron tomography and image analysis of intact Borrelia burgdorferi to produce a three-dimensional (3-D) model of the in situ flagellar motor without imposing rotational symmetry. Structural details of B. burgdorferi, including a layer of outer surface proteins, were clearly visible in the resulting 3-D reconstructions. By averaging the 3-D images of approximately 1,280 flagellar motors, a approximately 3.5-nm-resolution model of the stator and rotor structures was obtained. flgI transposon mutants lacked a torus-shaped structure attached to the flagellar rod, establishing the structural location of the spirochetal P ring. Treatment of intact organisms with the nonionic detergent NP-40 resulted in dissolution of the outermost portion of the motor structure and the C ring, providing insight into the in situ arrangement of the stator and rotor structures. Structural elements associated with the stator followed the curvature of the cytoplasmic membrane. The rotor and the C ring also exhibited angular flexion, resulting in a slight narrowing of both structures in the direction perpendicular to the cell axis. These results indicate an inherent flexibility in the rotor-stator interaction. The FliG switching and energizing component likely provides much of the flexibility needed to maintain the interaction between the curved stator and the relatively symmetrical rotor/C-ring assembly during flagellar rotation.


Somatic diversification and selection of immunoglobulin heavy and light chain variable region genes in IgG+ CD5+ chronic lymphocytic leukemia B cells.
Chronic lymphocytic leukemia (CLL) is characterized by the clonal expansion of CD5-expressing B lymphocytes. Most studies have found that these leukemic CD5+ B cells, like their normal counterparts, use immunoglobulin (Ig) variable (V) region genes that exhibit minimal, if any, somatic diversity. These and other observations have suggested that CD5+ B cells may be incapable of generating Ig V gene diversity, and therefore may not be able to develop higher affinity binding sites that could be selected by antigen. However, most of the studies of CLL and normal CD5+ B cells have focused on IgM-producing cells. Since somatic mutations are most often seen in B cells that have undergone an isotype class switch, we analyzed the Ig heavy (H) and light (L) chain variable region genes of seven IgG+CD5+ CLL B cells to determine if somatic diversification and antigen selection had occurred. The data derived provide evidence for skewed use, somatic diversification, and antigenic selection of the Ig V region genes. Nonrandom use of both H and L chain V region genes was manifested by an overrepresentation of VH4 and VKI family genes and the underrepresentation of the JH4 gene segment. Furthermore, VH4 gene use was restricted to only two family members (4.21 and 4.18). In four of the seven cases, the VH and VL genes displayed > or = 5% difference from the most homologous known germline counterparts. Polymerase chain reaction and Southern blot analyses performed in two of these patients demonstrated that their unique VH CDR2 and adjacent sequences were not present in their germline DNA. In addition, a significant level of diversity was seen in the rearranged DJH segments and at the VL-JL junctions of every patient that occurred both at the time of recombination and subsequently. The localization of replacement changes to complementarity determining regions of some patients suggested that antigen selection had occurred. Furthermore, the mutations identified in the VH and VL genes of each individual patient were strikingly similar, both in number and location. Collectively, the data indicate that a subset of CD5+ CLL B cells can display Ig V region gene mutations. In addition, they are consistent with the notions that in some cases antigen selection of these mutations may have occurred, and that antigen stimulation may be a promoting factor in the evolution of certain CLL clones.


Alterations in gene expression in sorghum induced by the host-specific toxin from Periconia circinata.
Susceptibility of sorghum to the fungal pathogen Periconia circinata and sensitivity to its host-specific toxin are determined by the semidominant allele at the pc locus. Pretreatment of susceptible seedlings with cycloheximide or cordycepin for 4 hr before treatment with the toxin protected the seedlings against toxin-induced loss of electrolytes and prevented development of disease symptoms. In vivo incorporation of [(3)H]leucine into protein was inhibited 91% and 47% by cycloheximide and cordycepin, respectively, but was not affected by the toxin. Gel electrophoresis and fluorography of in vivo-labeled proteins extracted from non-treated and toxin-treated root tips of near-isogenic susceptible and resistant lines revealed a selective increase in radioactivity of a protein band at M(r) 16,000 only in preparations from toxin-treated susceptible root tips. Two-dimensional gel electrophoresis separated the M(r) 16,000 band into four proteins and confirmed the increased rate of synthesis. Products of in vitro translation were substantially enriched with the four M(r) 16,000 proteins when total RNA from toxin-treated susceptible root tips was used in a cell-free protein-synthesizing system. Because the proteins that increase are common to both susceptible and resistant genotypes, the toxin apparently interferes with a regulatory function, perhaps a function of the pc locus, and thereby alters gene expression in the susceptible genotype. The data suggest but do not establish that phytotoxicity results from the increased rate of synthesis of the specific proteins.


Ryanodine receptors in human bladder smooth muscle.
The role of intracellular Ca2+ release in the activation of human bladder smooth muscle is controversial. We have measured the expression of mRNA encoding for the ryanodine receptor (RyR) isoforms (RyR1, RyR2 and RyR3) in isolated human detrusor smooth muscle. mRNA for RyR2 was detected in all samples but no mRNA for RyR1 or RyR3 could be found. Human bladder smooth muscle cells in culture are unresponsive to caffeine, suggesting the absence of a functional RyR system. However, mRNA encoding for RyR2 was detected in these cells. Using saponin-permeabilized cells, a Ruthenium Red-sensitive Ca(2+)-dependent 45Ca2+ release could be demonstrated from the sarcoplasmic reticulum (SR). These data confirm the functional presence of Ca(2+)-induced Ca2+ release (CICR) in cells and suggest that the properties of the RyR2 isoform in human detrusor may change when the cells are maintained in culture. The implications of these observations to detrusor smooth muscle function are discussed.


Categorical color constancy for simulated surfaces.
Color constancy is the ability to perceive constant surface colors under varying lighting conditions. Color constancy has traditionally been investigated with asymmetric matching, where stimuli are matched over two different contexts, or with achromatic settings, where a stimulus is made to appear gray. These methods deliver accurate information on the transformations of single points of color space under illuminant changes, but can be cumbersome and unintuitive for observers. Color naming is a fast and intuitive alternative to matching, allowing data collection from a large portion of color space. We asked observers to name the colors of 469 Munsell surfaces with known reflectance spectra simulated under five different illuminants. Observers were generally as consistent in naming the colors of surfaces under different illuminants as they were naming the colors of the same surfaces over time. The transformations in category boundaries caused by illuminant changes were generally small and could be explained well with simple linear models. Finally, an analysis of the pattern of naming consistency across color space revealed that largely the same hues were named consistently across illuminants and across observers even after correcting for category size effects. This indicates a possible relationship between perceptual color constancy and the ability to consistently communicate colors.


Incidence of mesothelioma in Lombardy, Italy: exposure to asbestos, time patterns and future projections.
In Italy, asbestos has been extensively used from 1945 to 1992. We evaluated the impact of exposure to asbestos on occurrence of malignant mesothelioma (MM) in the Lombardy Region, Northwest Italy, the most populated and industrialised Italian region. From the Lombardy Mesothelioma Registry, we selected all incident cases of MM diagnosed between 2000 and 2012. We described sources of exposure to asbestos and examined time trends of MM rates. Using Poisson age-cohort models, we derived projections of burden of MM in the Lombardy population for the period 2013-2029. In 2000-2012, we recorded 4442 cases of MM (2850 men, 1592 women). Occupational exposure to asbestos was more frequent in men (73.6%) than in women (38.2%). Non-occupational exposure was found for 13.6% of women and 3.6% of men. The average number of cases of MM per year was still increasing (+3.6% in men, +3.3% in women). Incidence rates were still increasing in individuals aged 65+ years and declining in younger people. A maximum of 417 cases of MM (267 men, 150 women) are expected in 2019. We forecast there will be 6832 more cases (4397 in men, 2435 in women) in the period 2013-2029, for a total of 11 274 cases of MM (7247 in men, 4027 in women) in 30 years. This study documented a high burden of MM in both genders in the Lombardy Region, reflecting extensive occupational (mainly in men) and non-occupational (mainly in women) exposure to asbestos in the past. Incidence rates are still increasing; a downturn in occurrence of MM is expected to occur after 2019.


Incidence of hibernating myocardium after acute myocardial infarction treated with thrombolysis.
To establish the incidence of hibernating myocardium after myocardial infarction treated with thrombolysis and to observe differences in the clinical outcome between patients with and without hibernating tissue. 41 patients underwent gated positron emission tomography with 18-fluorodeoxyglucose and 13N-ammonia at a median of eight days after first myocardial infarction. All 41 subjects had a matched perfusion-metabolism deficit in the region of myocardium indicated as the site of infarction by an electrocardiograph; 32 patients (78%) had scans which also showed at least one area of reduced blood flow and contraction with a concomitant increase in glucose uptake, representing hibernating myocardium. Patients were followed up at a median of six months: all 41 were alive and none had sustained a further infarct or cardiac arrhythmia; 17 subjects with hibernating tissue (53.1%) and two without (25%) reported chest pain after myocardial infarction. Hibernating myocardium is relatively common shortly after myocardial infarction treated with thrombolysis. It does not influence mortality or the incidence of postinfarction chest pain.


Service system involvement and delinquent offending at system of care entry.
This study examines correlates of delinquent offending among 2,554 youths presenting to community-based treatment in Center for Mental Health Services-funded systems of care. Variables in five key domains, including demographics, family risk, child clinical risk, school, and service system involvement, were examined in relation to severity of offending at treatment entry for early/middle (11-15 years) versus late (16-18 years) adolescents. Significant correlates of offending severity were identified in all domains except family risk for the 11 to 15 year olds and in demographics and child clinical risk for the 16 to 18 year olds. Service system involvement was a unique correlate of delinquency in the younger group. Findings add to other studies showing that correlates of offending appear to differ across stages of adolescence; specifically, school and service system involvement may be less important for older adolescents than for younger adolescents. Service system involvement should be included in longitudinal studies of risk for adolescent offending.


The California Tobacco Control Program's effect on adult smokers: (2) Daily cigarette consumption levels.
To investigate the association of the California Comprehensive Tobacco Control Program with self-reported population trends of cigarette consumption during 1992-2002. Participants were non-Hispanic white daily smokers (aged 20-64 years, n = 24 317) from the Tobacco Use Supplements to the Current Population Survey (1992-2002). We compared age-specific trends in consumption among daily smokers in three groups of states with differing tobacco control initiatives: California (CA; high cigarette price/comprehensive programme), New York and New Jersey (high cigarette price/no comprehensive programme), and tobacco-growing states (TGS; low cigarette price/no comprehensive programme). There was a general decline in cigarette consumption across all age groups in each category of states between 1992 and 2002, except the oldest age group in the TGS. The largest annual decline in the average number of cigarettes per day was observed among daily smokers in CA who were aged > or = 35 years (-0.41 cigarettes/day/year (95% CI -0.52 to -0.3)). This rate was significantly higher than the -0.22 cigarettes/day/year (95% CI -0.3 to -0.16; p<0.02) observed in same-age daily smokers from New York and New Jersey, and significantly higher than the rate in same-age daily smokers from the TGS (-0.15 cigarettes/day/year (95% CI -0.22 to -0.08; p<0.002)). There were no significant differences across state groups in the decline observed in daily smokers aged 20-34 years. In 2002, only 12% of daily smokers in CA smoked more than a pack per day, which was significantly lower than the 17% in New York and New Jersey, which again was significantly lower than the 25% in the TGS. The California Tobacco Control Program was associated with significant declines in cigarette consumption among daily smokers aged > or = 35 years of age, which in turn should lead to declines in tobacco-related health effects. The decline in consumption among young adult smokers was a national trend.


Sexual violence against migrants and asylum seekers. The experience of the MSF clinic on Lesvos Island, Greece.
Sexual violence can have a destructive impact on the lives of people. It is more common in unstable conditions such as during displacement or migration of people. On the Greek island of Lesvos, Médecins Sans Frontières provided medical care to survivors of sexual violence among the population of asylum seekers. This study describes the patterns of sexual violence reported by migrants and asylum seekers and the clinical care provided to them. This is a descriptive study, using routine program data. The study population consisted of migrants and asylum seekers treated for conditions related to sexual violence at the Médecins Sans Frontières clinic on Lesvos Island (September 2017-January 2018). There were 215 survivors of sexual violence who presented for care, of whom 60 (28%) were male. The majority of incidents reported (94%) were cases of rape; 174 (81%) of survivors were from Africa and 185 (86%) of the incidents occurred over a month before presentation. Half the incidents (118) occurred in transit, mainly in Turkey, and 76 (35%) in the country of origin; 10 cases (5%) occurred on Lesvos. The perpetrator was known to the survivor in 23% of the cases. The need for mental health care exceeded the capacity of available mental care services. Even though the majority of cases delayed seeking medical care after the incident, it is crucial that access to mental health services is guaranteed for those in need. Such access and security measures for people in transit need to be put in place along migration routes, including in countries nominally considered safe, and secure routes need to be developed.


The potential of crowdsourcing to improve patient-centered care.
Crowdsourcing (CS) is the outsourcing of a problem or task to a crowd. Although patient-centered care (PCC) may aim to be tailored to an individual's needs, the uses of CS for generating ideas, identifying values, solving problems, facilitating research, and educating an audience represent powerful roles that can shape both allocation of shared resources and delivery of personalized care and treatment. CS can often be conducted quickly and at relatively low cost. Pitfalls include bias, risks of research ethics, inadequate quality of data, inadequate metrics, and observer-expectancy effect. Health professionals and consumers in the US should increase their attention to CS for the benefit of PCC. Patients' participation in CS to shape health policy and decisions is one way to pursue PCC itself and may help to improve clinical outcomes through a better understanding of patients' perspectives. CS should especially be used to traverse the quality-cost curve, or decrease costs while preserving or improving quality of care.


Racial disparities in the development of breast cancer metastases among older women: a multilevel study.
Distant metastases are the most common and lethal type of breast cancer relapse. The authors examined whether older African American breast cancer survivors were more likely to develop metastases compared with older white women. They also examined the extent to which 6 pathways explained racial disparities in the development of metastases. The authors used 1992-1999 Surveillance, Epidemiology, and End Results (SEER) data with 1991-1999 Medicare data. They used Medicare's International Classification of Diseases, Ninth Revision, Clinical Modification codes to identify metastases of respiratory and digestive systems, brain, bone, or other unspecified sites. The 6 pathways consisted of patient characteristics, tumor characteristics, type of treatment received, access to medical care, surveillance mammography use, and area-level characteristics (poverty rate and percentage African American) and were obtained from the SEER or Medicare data. Of the 35,937 women, 10.5% developed metastases. In univariate analysis, African American women were 1.61 times (95% confidence interval [CI], 1.54-1.83) more likely to develop metastasis than white women. In multivariate analysis, tumor grade, stage at diagnosis, and census-tract percentage African American explained why African American women were more likely to develop metastases than white women (hazard ratio, 0.84; 95% CI, 0.68-1.03). Interventions to reduce late-stage breast cancer among African Americans also may reduce racial disparities in subsequent increased risk of developing metastasis. African Americans diagnosed with high-grade breast cancer could be targeted to reduce their risk of metastasis. Future studies should identify specific reasons why the racial distribution in census tracts was associated with racial disparities in the risk of breast cancer metastases.


One-year results of the Thrombolysis in Myocardial Infarction (TIMI) IIIB clinical trial. A randomized comparison of tissue-type plasminogen activator versus placebo and early invasive versus early conservative strategies in unstable angina and non-Q wave myocardial infarction.
We report mortality, infarction, revascularization and repeat hospital admission events for 1 year after enrollment and randomization in the Thrombolysis in Myocardial Ischemia (TIMI) IIIB clinical trial. The purpose of this trial was to investigate the role of a thrombolytic agent added to conventional medical therapies and to compare an early invasive management strategy to a more conservative early strategy in patients with unstable angina and non-Q wave myocardial infarction. There were 1,473 patients enrolled, and they received conventional anti-ischemic medical therapies. They were randomized to therapy with either tissue-type plasminogen activator (t-PA) or placebo and also to an early invasive management strategy with coronary arteriography at 18 to 48 h, followed by revascularization as soon as possible if appropriate, or, alternatively, to an early conservative strategy with arteriography and revascularization reserved for failure of initial therapy to prevent recurrent ischemia. The primary end point was a composite outcome variable and was assessed at 42 days. Patients were then managed entirely at the discretion of their treating physician. Follow-up contacts were made at 1 year. The incidence of death or nonfatal infarction for the t-PA- and placebo-treated groups was similar after 1 year (12.4% vs. 10.6%, p = 0.24). The incidence of death or nonfatal infarction was also similar after 1 year for the early invasive and early conservative strategies (10.8% vs. 12.2%, p = 0.42). A trial of this size should be able to detect differences in relative risk for death or infarction > or = 1.81 with a power of 80% at a significance level (alpha) of 0.01. Revascularization by 1 year was common, but was slightly more common with the early invasive than the early conservative strategy (64% vs. 58%, p < 0.001). This result was related entirely to a small difference in angioplasty rates (39% vs. 32%, p < 0.001) inasmuch as rates of bypass grafting by 1 year were equivalent (30% in each group, p = 0.50). The high rate of revascularization in both strategies was accompanied by comparable clinical status at the 1-year follow-up contact. In this large study of unstable angina and non-Q wave myocardial infarction, the incidence of death and nonfatal infarction or reinfarction was low but not trivial after 1 year (4.3% mortality, 8.8% nonfatal infarction). An early invasive management strategy was associated with slightly more coronary angioplasty procedures but equivalent numbers of bypass surgery procedures than a more conservative early strategy of catheterization and revascularization only for signs of recurrent ischemia. The incidence of death or nonfatal infarction, or both, did not differ after 1 year by strategy assignment, but fewer patients in the early invasive strategy group underwent later repeat hospital admission (26% vs. 33%, p < 0.001). Either strategy is appropriate for patient management; differences in hospital admissions and revascularization procedures, with their attendant costs, are likely to be minimal.


Effectiveness of percutaneous posterior tibial nerve stimulation for overactive bladder: a systematic review and meta-analysis.
To evaluate the effectiveness of posterior percutaneous tibial nerve stimulation (PTNS) in treating overactive bladder (OAB) symptoms by systematic review of the literature. Systematic literature search was carried out (up to April 2011) using relevant search terms in Medline, EMBASE, CINAHL, CENTRAL, National Library for Health, MetaRegister of controlled trials, LILACS, and Google Scholar. Relevant randomized controlled trials (RCTs) and prospective studies were selected and then analyzed by two-independent reviewers. Meta-analysis was performed with random effects model using STATA 8 for non-randomized prospective studies and with Review Manager 5.1 for RCTs. The studies report variable initial success rates (37-82%) for treating OAB symptoms with PTNS. Four randomized trials compared PTNS with Sham treatment showing a significant difference favoring PTNS [RR 7.02 95% confidence interval (CI) 1.69-29.17]. Two randomized trials compared PTNS with antimuscarinic medication with no significant difference in the change in bladder diary parameters between the treatments. Ten prospective non-randomized studies were included. The definitions of success were varied. The pooled subjective success rate was 61.4% (95% CI 57.5-71.8) and objective success rate was 60.6% (95% CI 49.2-74.7). There is evidence of significant improvement in OAB symptoms using PTNS which is comparable to the effect of antimuscarinics but with a better side effect profile. The studies included in the review only considered short-term outcomes after initial treatment. In order to recommend PTNS as a practical treatment option, long-term data and health economic analysis are needed.


Mutations in U4atac snRNA, a component of the minor spliceosome, in the developmental disorder MOPD I.
Small nuclear RNAs (snRNAs) are essential factors in messenger RNA splicing. By means of homozygosity mapping and deep sequencing, we show that a gene encoding U4atac snRNA, a component of the minor U12-dependent spliceosome, is mutated in individuals with microcephalic osteodysplastic primordial dwarfism type I (MOPD I), a severe developmental disorder characterized by extreme intrauterine growth retardation and multiple organ abnormalities. Functional assays showed that mutations (30G>A, 51G>A, 55G>A, and 111G>A) associated with MOPD I cause defective U12-dependent splicing. Endogenous U12-dependent but not U2-dependent introns were found to be poorly spliced in MOPD I patient fibroblast cells. The introduction of wild-type U4atac snRNA into MOPD I cells enhanced U12-dependent splicing. These results illustrate the critical role of minor intron splicing in human development.


Screening For Gestational Diabetes Mellitus Selectively in a University Maternity Hospital
Aims Screening for gestational diabetes mellitus (GDM) may be universal or selective based on risk factors. We audited selective screening with an Oral Glucose Tolerance Test (OGTT). Methods Clinical and laboratory details of the first 200 women who delivered a baby in 2017 were analysed. Results Based on national recommendations, 46.5% (n=93) had maternal risk factors (RF) and an additional 6.5% (n=13) had fetal RF. Nine women with RF, for unexplained reasons did not have their OGTT. Of the 95 who had their OGTT, the diagnosis of GDM was made in 27.4% (n=26). The diagnosis of GDM was made in an additional 8 women outside selective screening giving an overall incidence of 17.0%. Discussion More than half of the women needed to be screened selectively for GDM. Compliance with the national recommendations was incomplete and thus the diagnosis of GDM may be missed even in an academic setting.


Technical note: ruminal cannulation technique in young Holstein calves: effects of cannulation on feed intake, body weight gain, and ruminal development at six weeks of age.
Ruminal cannulation techniques are frequently used to study fermentation in the ruminant forestomach. Unsatisfactory results with the traditionally applied procedure for cannulation of young calves stimulated the development of a simpler and more robust procedure; this procedure was tested for effects on performance traits and gross anatomy of the gastrointestinal tract compared with a control group not undergoing surgery. Five calves were ruminally cannulated at approximately 10 d of age and 5 matching calves were used as controls. All calves were fed milk replacer and a diet based on clover grass silage and sodium hydroxide-treated wheat. Ruminal fluid was collected from cannulated calves once weekly for 3 consecutive weeks. All calves were euthanized at 43+/-3 d of age. No apparent adverse effects of cannulation were observed. Feed intake, BW gain, and gross anatomy of the gastrointestinal tract were not affected by cannulation. Minimum ruminal pH increased with sampling week, but average ruminal pH, total volatile fatty acids concentration, and volatile fatty acids proportions were not affected by sampling week. In conclusion, the implemented surgical technique was found to have no major effect on apparent animal health and performance traits, and the cannula proved useful for multiple samplings of ruminal contents in young calves.


Genetic and transcriptional analysis of human host response to healthy gut microbiota.
Many studies have demonstrated the importance of the gut microbiota in healthy and disease states. However, establishing the causality of host-microbiota interactions in humans is still challenging. Here, we describe a novel experimental system to define the transcriptional response induced by the microbiota in human cells and to shed light on the molecular mechanisms underlying host-gut microbiota interactions. In primary human colonic epithelial cells, we identified over 6,000 genes that change expression at various time points following co-culturing with the gut microbiota of a healthy individual. Among the differentially expressed genes we found a 1.8-fold enrichment of genes associated with diseases that have been previously linked to the microbiome, such as obesity and colorectal cancer. In addition, our experimental system allowed us to identify 87 host SNPs that show allele-specific expression in 69 genes. Furthermore, for 12 SNPs in 12 different genes, allele-specific expression is conditional on the exposure to the microbiota. Of these 12 genes, eight have been associated with diseases linked to the gut microbiota, specifically colorectal cancer, obesity and type 2 diabetes. Our study demonstrates a scalable approach to study host-gut microbiota interactions and can be used to identify putative mechanisms for the interplay between host genetics and microbiota in health and disease.


Conditional deletion of Smad1 and Smad5 in somatic cells of male and female gonads leads to metastatic tumor development in mice.
The transforming growth factor beta (TGFbeta) family has critical roles in the regulation of fertility. In addition, the pathogenesis of some human cancers is attributed to misregulation of TGFbeta function and SMAD2 or SMAD4 mutations. There are limited mouse models for the BMP signaling SMADs (BR-SMADs) 1, 5, and 8 because of embryonic lethality and suspected genetic redundancy. Using tissue-specific ablation in mice, we deleted the BR-SMADs from somatic cells of ovaries and testes. Single conditional knockouts for Smad1 or Smad5 or mice homozygous null for Smad8 are viable and fertile. Female double Smad1 Smad5 and triple Smad1 Smad5 Smad8 conditional knockout mice become infertile and develop metastatic granulosa cell tumors. Male double Smad1 Smad5 conditional knockout mice are fertile but demonstrate metastatic testicular tumor development. Microarray analysis indicated significant alterations in expression of genes related to the TGFbeta pathway, as well as genes involved in infertility and extracellular matrix production. These data strongly implicate the BR-SMADs as part of a critical developmental pathway in ovaries and testis that, when disrupted, leads to malignant transformation.


Workshop summary: Potential usefulness and feasibility of a US National Mesothelioma Registry.
The burden and prognosis of malignant mesothelioma in the United States have remained largely unchanged for decades, with approximately 3200 new cases and 2400 deaths reported annually. To address care and research gaps contributing to poor outcomes, in March of 2019 the Mesothelioma Applied Research Foundation convened a workshop on the potential usefulness and feasibility of a national mesothelioma registry. The workshop included formal presentations by subject matter experts and a moderated group discussion. Workshop participants identified top priorities for a registry to be (a) connecting patients with high-quality care and clinical trials soon after diagnosis, and (b) making useful data and biospecimens available to researchers in a timely manner. Existing databases that capture mesothelioma cases are limited by factors such as delays in reporting, deidentification, and lack of exposure information critical to understanding as yet unrecognized causes of disease. National disease registries for amyotrophic lateral sclerosis (ALS) in the United States and for mesothelioma in other countries, provide examples of how a registry could be structured to meet the needs of patients and the scientific community. Small-scale pilot initiatives should be undertaken to validate methods for rapid case identification, develop procedures to facilitate patient access to guidelines-based standard care and investigational therapies, and explore approaches to data sharing with researchers. Ultimately, federal coordination and funding will be critical to the success of a National Mesothelioma Registry in improving mesothelioma outcomes and preventing future cases of this devastating disease.


Unique insight into microenvironmental changes in colorectal cancer: Ex vivo assessment of matrix metalloprotease-mediated molecular changes in human colorectal tumor tissue and corresponding non-neoplastic adjacent tissue.
Matrix metalloprotease (MMP)-mediated tissue remodeling is one of the malignant changes driving colorectal cancer. Measurement of altered MMP expression/activity is not sufficient to fully understand the effect of MMP-mediated tissue remodeling. Biomarkers are required that specifically reflect the dynamic processes of the MMP-mediated degradation of signature proteins from colorectal tissue. The aim of the present study was to profile and characterize the release of MMP-degraded type III collagen (C3M) and citrullinated and MMP-degraded vimentin (VICM) from tumor tissue and corresponding non-neoplastic adjacent tissue (NAT) in a human colorectal cancer ex vivo model. Colorectal tumor tissue and NAT biopsies from tissue removed during resection of colorectal cancer patients (n=13) were cut into pieces of 2 mm2 and cultured for 24 h in growth medium. C3M and VICM were evaluated by ELISA. As part of the characterization, C3M was determined subsequent to the tumor tissue being cleaved with recombinant MMP-2/-9 and trypsin. C3M was generated by MMP-2/-9, but not by trypsin. In addition, the level of C3M was significantly elevated in the conditioned medium from tumor tissues (3.7 ng/ml) compared with that observed in the conditioned medium from the NATs (2.2 ng/ml) and in the growth medium alone (1.9 ng/ml). The level of VICM was significantly elevated in the tumor tissues (0.51 ng/ml) and NATs (0.52 ng/ml) compared with that in the growth medium alone (0.03 ng/ml). No differences were detected between the tumor tissues and NATs. No correlation was observed between biomarker levels from the tumor tissue and corresponding NAT, and the biomarker levels did not correlate with tumor stage. In conclusion, the present study provided support of the concept that C3M and VICM are applicable as tools to investigate dynamic tissue changes of colorectal tumor tissue and corresponding NAT. By the assessment of these specific MMP-mediated molecular changes, the present study provides novel and relevant insight into the dynamic changes of colorectal tumor tissue and corresponding NAT.


Cyclooxygenase-1-dependent prostaglandins mediate susceptibility to systemic inflammation-induced acute cognitive dysfunction.
Systemic inflammatory events often precipitate acute cognitive dysfunction in elderly and demented populations. Delirium is a highly prevalent neuropsychiatric syndrome that is characterized by acute inattention and cognitive dysfunction, for which prior dementia is the major predisposing factor and systemic inflammation is a frequent trigger. Inflammatory mechanisms of delirium remain unclear. We have modeled aspects of delirium during dementia by exploiting progressive neurodegeneration in the ME7 mouse model of prion disease and by superimposing systemic inflammation induced by the bacterial endotoxin lipopolysaccharide (LPS). Here, we have used this model to demonstrate that the progression of underlying disease increases the incidence, severity, and duration of acute cognitive dysfunction. This increasing susceptibility is associated with increased CNS expression of cyclooxygenase (COX)-1 in microglia and perivascular macrophages. The COX-1-specific inhibitor SC-560 provided significant protection against LPS-induced cognitive deficits, and attenuated the disease-induced increase in hippocampal and thalamic prostaglandin E2, while the COX-2-specific inhibitor NS-398 was ineffective. SC-560 treatment did not alter levels of the proinflammatory cytokines interleukin (IL)-1β, tumor necrosis factor-α, IL-6, or C-X-C chemokine ligand 1 in blood or brain, but systemic IL-1RA blocked LPS-induced cognitive deficits, and systemic IL-1β was sufficient to induce similar deficits in the absence of LPS. Furthermore, the well tolerated COX inhibitor ibuprofen was protective against IL-1β-induced deficits. These data demonstrate that progressive microglial COX-1 expression and prostaglandin synthesis can underpin susceptibility to cognitive deficits, which can be triggered by systemic LPS-induced IL-1β. These data contribute to our understanding of how systemic inflammation and ongoing neurodegeneration interact to induce cognitive dysfunction and episodes of delirium.


An alternative in vivo system for testing angiogenic potential of human neuroblastoma cells.
In this study we purposed an alternative method to study the angiogenic and invasive potential of neuroblastoma cell suspensions implanted on the chick embryo chorioallantoic membrane (CAM) surface. Neuroblastoma cells were seeded in Matrigel and thereafter the suspension was pipetted onto the CAM surface at day 8 of incubation inside a silicon ring previously loaded onto the CAM surface. Four days after implantation, the silicon ring was removed and the angiogenic and invasive response were studied morphologically at macroscopic and microscopic levels and by reverse transcriptase-polymerase chain reaction (RT-PCR) by using human and chicken primers for several angiogenic cytokines, namely vascular endothelial growth factor-A (VEGF-A), fibroblast growth factor-2 (FGF-2), angiopoietin-1 (ANG-1), hypoxia inducible factor-2alpha (HIF-2alpha), and for an endogenous angiostatic molecule, namely endostatin. Results showed that: (1) Neuroblastoma cells induced an angiogenic response in the CAM assay comparable to that induced by FGF-2; (2) neuroblastoma cells are packed inside Matrigel or are recognizable in the CAM mesenchyme; (3) Angiogenic activity of neuroblastoma cells is associated to an high expression of the transcripts of human VEGF-A, FGF-2, ANG-1 and HIF-2alpha and to a low expression in the transcript of a human endostatin while in the control specimens there is no expression of both angiogenic and angiostatic molecules; and (4) the expression of the transcripts of the same chicken angiogenesis stimulators and inhibitor is unmodified in treated and control specimens. Overall, these data indicate that neuroblastoma cells growth on the chick CAM express characteristics of the human disease. This experimental model could be employed for further research on human tumor progression and anti-angiogenic molecules screening.


Seasonality of mortality and graft failure among kidney transplant recipients in the US - a retrospective study.
Mortality in the general population and in patients on chronic hemodialysis is significantly higher in winter than summer. It is unknown whether such a seasonal difference exists for mortality or graft failure among kidney transplant recipients. We analyzed United Network for Organ Sharing (UNOS) data to assess whether the annual distribution of deaths and graft failures differed significantly from expected. There was significant annual variation in both deaths (n = 52 523) and graft failures (n = 50 301; both P < 0.001). The number of observed deaths exceeded the number expected by 8.9% in winter (P < 0.001), whereas the number of deaths was 4.8% lower than expected in summer (P < 0.01). The pattern was strongest for deaths attributable to cardiovascular disease (n = 11 509; 21.9%). Similarly, there was an excess of graft failures in winter (3.6%; P < 0.01) and a deficit in other seasons (all P ≤ 0.02). This pattern was observed for graft failures due to chronic rejection (P < 0.001) and other causes (P < 0.001), but not for acute rejection (P = 0.28) or recurrent disease (P = 0.27). Potential explanations for this variation include changes in physiologic parameters, changes in medication adherence and other behaviors, or changes in insurance coverage or clinical care. Further studies are necessary to identify specific mechanisms.


Biomonitoring of smoke constituents: exposure to 4-aminobiphenyl and 4-aminobiphenyl hemoglobin adduct levels in nonsmokers and smokers.
Public health authorities worldwide have concluded that exposure to environmental tobacco smoke (ETS) causes diseases, including cancer, in adult nonsmokers. The arylamine, 4-aminobiphenyl (4-ABP), has been identified as a human carcinogen. Some publications have suggested that 4-ABP hemoglobin (4-ABP-Hb) adduct levels in nonsmokers are a result of exposure to ETS, whereas others could not confirm these observations. Toxicokinetic and exposure models proposed in this work are used to estimate the concentration of 4-ABP-Hb adducts resulting from ETS exposure that is based on experimental values for respirable suspended particulates (RSP) concentration. Monte Carlo methods were used to obtain estimates of population distributions of 4-ABP-Hb adduct levels resulting from indoor ETS exposure in homes, workplaces, and hospitality environments. It is found that the mean, median, and 95th percentile 4-ABP-Hb adduct steady-state levels of 0.4-1.4, 0.2-1.0, and 0.97-4.63 pg/g Hb, respectively, are estimated from ETS exposure. These 4-ABP-Hb adduct levels from ETS exposure account for approximately 1-4% of the median levels reported for nonsmokers, explaining, in part, contradictory literature data on 4-ABP-Hb adduct levels in nonsmokers. No risk assessment of ETS or 4-ABP was conducted in this work, consequently the known health effects of ETS are neither confirmed or challenged and our conclusions are limited to the determination that ETS is not a major source of 4-ABP-Hb adduct levels in non-smokers.


Freshwater reservoir offsets on radiocarbon-dated dog bone from the headwaters of the St. Lawrence River, USA.
Isotopic analysis of dog (Canis lupus familiaris) bone recovered from archaeological sites as proxies for human bone is becoming common in North America. Chronological placement of the dogs is often determined through radiocarbon dating of dog bone. The Great Lakes, their tributaries, and nearby lakes and streams were important fisheries for Native Americans prior to and after sustained European presence in the region. Carbon entering the food web in freshwater systems is often not in full isotopic equilibrium with the atmosphere, giving rise to spuriously old radiocarbon ages in fish, other aquatic organisms, and their consumers. These freshwater reservoir offsets (FROs) have been noted on human and dog bone in several areas of the world. Here we report the results of multi-tracer Bayesian dietary modeling using δ15N and δ13C values on dog bone collagen from mid-fifteenth to mid-sixteenth-century Iroquoian village sites at the headwaters of the St. Lawrence River, New York, USA. Results indicate that fish was an important component of dog diets. A comparison of radiocarbon dates on dog bone with dates on deer bone or maize from the same sites indicate FROs ranging from 97 ± 24 to 220 ± 39 14Cyr with a weighted mean of 132 ± 8 14Cyr. These results suggest that dog bone should not be used for radiocarbon dating in the absence of modeling to determine fish consumption and that previously reported radiocarbon dates on human bone from the larger region are likely to have FROs given the known importance of fish in regional human diets.


Experimental identification of social learning in wild animals.
Field experiments can provide compelling demonstrations of social learning in wild populations. Social learning has been experimentally demonstrated in at least 23 field experiments, in 20 species, covering a range of contexts, such as foraging preferences and techniques, habitat choice, and predator avoidance. We review experimental approaches taken in the field and with wild animals brought into captivity and note how these approaches can be extended. Relocating individuals, introducing trained individual demonstrators or novel behaviors into a population, or providing demonstrator-manipulated artifacts can establish whether and how a particular act can be socially transmitted in the wild and can help elucidate the benefits of social learning. The type, strength, and consistency of presented social information can be varied, and the provision of conditions favoring the performance of an act can both establish individual discovery rates and help determine whether social information is needed for acquisition. By blocking particular avenues of social transmission or removing key individuals, routes of transmission in wild populations can be investigated. Manipulation of conditions proposed to favor social learning can test mathematical models of the evolution of social learning. We illustrate how field experiments are a viable, vital, and informative approach to the study of social learning.


Mitochondrial Dysfunction and Chronic Disease: Treatment With Natural Supplements.
Loss of function in mitochondria, the key organelle responsible for cellular energy production, can result in the excess fatigue and other symptoms that are common complaints in almost every chronic disease. At the molecular level, a reduction in mitochondrial function occurs as a result of the following changes: (1) a loss of maintenance of the electrical and chemical transmembrane potential of the inner mitochondrial membrane, (2) alterations in the function of the electron transport chain, or (3) a reduction in the transport of critical metabolites into mitochondria. In turn, these changes result in a reduced efficiency of oxidative phosphorylation and a reduction in production of adenosine-5'-triphosphate (ATP). Several components of this system require routine replacement, and this need can be facilitated with natural supplements. Clinical trials have shown the utility of using oral replacement supplements, such as l-carnitine, alpha-lipoic acid (α-lipoic acid [1,2-dithiolane-3-pentanoic acid]), coenzyme Q10 (CoQ10 [ubiquinone]), reduced nicotinamide adenine dinucleotide (NADH), membrane phospholipids, and other supplements. Combinations of these supplements can reduce significantly the fatigue and other symptoms associated with chronic disease and can naturally restore mitochondrial function, even in long-term patients with intractable fatigue.


A Phenotypic Screening Assay Identifies Modulators of Diamond Blackfan Anemia.
Diamond-Blackfan anemia (DBA) is a bone marrow failure syndrome caused by mutations in ribosomal protein genes. Pathogenic mechanisms are poorly understood but involve severely reduced proliferation of erythroid precursors. Because current DBA therapies are ineffective and associated with severe side effects, disease-specific therapies are urgently needed. We hypothesized that druggable molecular pathways underlying the defect can be revealed through phenotypic small-molecule screens. Accordingly, a screening assay was developed using c-kit+ fetal liver erythroid progenitors from a doxycycline-inducible DBA mouse model. The addition of doxycycline to the culture medium induces the phenotype and reduces proliferation to <10% of normal, such that rescue of proliferation can be used as a simple readout for screening. Here, we describe the assay rationale and efforts toward validation of a microtiter plate-compatible assay and its application in a pilot screen of 3871 annotated compounds. Ten hits demonstrated concentration-dependent activity, and we report a brief follow-up of one of these compounds. In conclusion, we established a robust scalable assay for screening molecules that rescue erythropoiesis in DBA.


A lower viral set point but little immunological impact after early treatment during primary HIV infection.
The Primo-SHM trial, a multicenter randomized trial comparing no treatment with 24 or 60 weeks of combination antiretroviral therapy (cART) during primary human immunodeficiency virus (HIV) infection (PHI), recently demonstrated that temporary early cART lowered the viral set point and deferred the need for re-initiation of cART during chronic HIV infection. This study examined whether the beneficial effect of early treatment was caused by preservation of immunological responses. Twenty-seven treated and 20 untreated PHI individuals participating in the Primo-SHM trial were compared at viral set point, that is, 36 weeks after baseline or after treatment interruption, respectively, for a diverse set of immunological parameters. The results show no differences between treated and untreated individuals at the level of effector T-cell formation or replication capacity of the T-cells; regulation of various T, B, natural killer, or dendritic cells; polyfunctionality of the CD8 T-cells; preservation of CD4 T-cells in the gut associated lymphoid tissue; or immune activation. There were subtle differences in the quality of the cytolytic CD4 T-cell response: 11% (median) of CD4 T-cells of the early treated individuals produced the cytolytic molecule perforin compared to 5% in untreated individuals (p=0.046), and treatment caused a modest increase in CD4 T-cells expressing both perforin and granzyme B (median 9% vs. 4% of CD4 T-cells; p=0.045). Early treatment had a modest positive effect on the quality of the CD4 T-cell response. It remains unclear, however, whether these subtle immunological differences were the cause or a result of the lower viral set point in patients who received early treatment.


Adhesive capsulitis: a new management protocol to improve passive range of motion.
To examine the short-term efficacy of a nonoperative shoulder protocol for the treatment of adhesive capsulitis. A retrospective chart review was used to collect data for a 3-year period. Academic tertiary medical center. 28 consecutive patients diagnosed as having adhesive capsulitis were identified and managed with a new protocol. The protocol consisted of the administration of a suprascapular nerve block, the subsequent injection of an intra-articular steroid, and then the injection of an anesthetic agent with brisement normal saline volume dilation. The final step was manipulation of the shoulder. A paired t test was used to examine the difference in the preprocedure and postprocedure passive range of motion (flexion and abduction). The average shoulder abduction before the procedure was 89.5 degrees ; this improved by an average of 51.7 degrees (P<.0001). The average shoulder flexion improved from 117.3 degrees by an average of 37.7 degrees (P<.0001). There was no significant difference in either abduction or flexion based on age, gender, or chronicity of symptoms. White patients experienced significantly more improvement in abduction than did nonwhite patients. This study suggests that this adhesive capsulitis management protocol is effective and produces a significant improvement in the passive range of motion immediately after the procedure.


Telomerase Repairs Collapsed Replication Forks at Telomeres.
Telomeres are difficult-to-replicate sites whereby replication itself may threaten telomere integrity. We investigate, in fission yeast, telomere replication dynamics in telomerase-negative cells to unmask problems associated with telomere replication. Two-dimensional gel analysis reveals that replication of telomeres is severely impaired and correlates with an accumulation of replication intermediates that arises from stalled and collapsed forks. In the absence of telomerase, Rad51, Mre11-Rad50-Nbs1 (MRN) complex, and its co-factor CtIPCtp1 become critical to maintain telomeres, indicating that homologous recombination processes these intermediates to facilitate fork restart. We further show that a catalytically dead mutant of telomerase prevents Ku recruitment to telomeres, suggesting that telomerase and Ku both compete for the binding of telomeric-free DNA ends that are likely to originate from a reversed fork. We infer that Ku removal at collapsed telomeric forks allows telomerase to repair broken telomeres, thereby shielding telomeres from homologous recombination.


Evaluation of Aspergillus PCR protocols for testing serum specimens.
A panel of human serum samples spiked with various amounts of Aspergillus fumigatus genomic DNA was distributed to 23 centers within the European Aspergillus PCR Initiative to determine analytical performance of PCR. Information regarding specific methodological components and PCR performance was requested. The information provided was made anonymous, and meta-regression analysis was performed to determine any procedural factors that significantly altered PCR performance. Ninety-seven percent of protocols were able to detect a threshold of 10 genomes/ml on at least one occasion, with 83% of protocols reproducibly detecting this concentration. Sensitivity and specificity were 86.1% and 93.6%, respectively. Positive associations between sensitivity and the use of larger sample volumes, an internal control PCR, and PCR targeting the internal transcribed spacer (ITS) region were shown. Negative associations between sensitivity and the use of larger elution volumes (≥100 μl) and PCR targeting the mitochondrial genes were demonstrated. Most Aspergillus PCR protocols used to test serum generate satisfactory analytical performance. Testing serum requires less standardization, and the specific recommendations shown in this article will only improve performance.


Pharmaceutical care of patients with cancer treated on non-cancer wards in the UK: a mixed methods study.
The objectives of this study were to identify and describe specialist cancer pharmacist support for the pharmaceutical care of patients with cancer treated on non-cancer wards. The study also explored the experiences and opinions of the pharmacy team concerning the pharmaceutical care of these patients. A mixed methods approach was used. A survey of the pharmaceutical care provided to patients with cancer treated on non-cancer wards across the UK was carried out. A focus group was conducted at a district general hospital. Main outcome measures included description of services provided by other Trusts from survey responses and themes from pharmacy staff experiences. Twenty two per cent of respondents answering the question (29/134) confirmed that pharmacy staff on the acute oncology team would review these patients. The most frequently reported descriptions of other cancer pharmacy support were provision of advice on an individual situation basis (83/132: 63%) and lack of resource to provide a service (25/132: 19%). The themes identified in the focus group discussion were: pharmaceutical care of patients, robust systems to support their care and provision of education and training. Participants expressed their dissatisfaction with the current service provision and lack of resources available to them. Specialist pharmacist advice was described as being available sometimes and participants recounted incidents in which they did not know what to do or lacked the knowledge to deal with themselves. The majority of respondents reported provision of ad hoc support from specialist cancer pharmacists for the pharmaceutical care of patients with cancer when they are treated on non-cancer wards. However, from their experiences generalist pharmacists would prefer to have more education and training to identify patients' pharmaceutical care needs in addition to ad hoc support.


Tethering tracts in spina bifida occulta: revisiting an established nomenclature.
The goal of the present study goal was to systematically confirm the previously recognized nomenclature for tethering tracts that are part of the spectrum of occult spinal dysraphic lesions. The tethering tract in 20 patients with spina bifida occulta underwent histological examination with H & E staining and epithelial membrane antigen (EMA) immunolabeling, and additional selected specimens were stained with Masson trichrome. All tethering tracts contained fibrous connective tissue. Four tracts were lined with epithelial cells and either originated within a dermoid cyst, terminated at a skin dimple/sinus opening, or had both of these characteristics. No tethering tracts exhibited EMA positivity or meningeal elements. Although all tethering tracts originated in juxtaposition to the spinal cord, their termination sites were variable. Based on histological findings and presumed embryological origin, the authors broadly classified tethering tracts terminating within the dura mater, epidural space, or lamina as "short tethering tracts" (STTs). The STTs occurred mostly in conjunction with split cord malformations and had a purely fibrous composition. Tethering tracts terminating superficial to the overlying lamina were classified as "long tethering tracts" (LTTs), and the authors propose that these are embryologically distinct from STTs. The LTTs were of two varieties: epithelial and nonepithelial, the former being typically associated with a skin dimple or spinal cord (epi)dermoid cyst. In fact, analysis of the data suggested that not every tethering tract terminating in or on the skin should be classified as a dermal sinus tract without histological confirmation, and because no evidence of meningeal tissue-lined tracts was detected, the use of the term "meningocele manqué" may not be appropriate.


Consumption of green tea favorably affects oxidative stress markers in weight-trained men.
This study investigated the effects of the consumption of green tea (GT) for 7 d on biomarkers of oxidative stress in young men undergoing resistance exercise. Fourteen subjects performed a bench press exercise (four sets, 10 to 4 repetitions) after undergoing a period without (control group) or with the intake of GT (GT group; 2 g of leaves in 200 mL of water, three times per day). Blood samples were obtained before and after exercise and analyzed for total antioxidant capacity (ferric reducing ability of plasma [FRAP]), total polyphenols, reduced glutathione (GSH), lipid hydroperoxide (LH) and thiobarbituric acid-reactive substances, creatine kinase (CK), aspartate aminotransferase (AST), xanthine oxidase (XO), hypoxanthine, and uric acid (UA). In the control group, exercise did not affect the values of LH, thiobarbituric acid-reactive substances, and FRAP, although it did reduce the levels of GSH (P < 0.05). In addition, exercise increased CK, AST, and XO activities, although it did not change the values for hypoxanthine or UA. Green tea reduced the postexercise concentration of LH and increased the values of total polyphenols, GSH, and FRAP. GT also inhibited a significant rise in CK and XO activities induced by exercise. Furthermore, GT decreased the AST activity and hypoxanthine and UA concentrations before and after exercise. The assessment of food consumption revealed that the participants had an unbalanced diet, particularly in relation to vitamin E and carotenoids. Consumption of GT, a beverage rich in polyphenols, may offer protection against the oxidative damage caused by exercise, and dietary guidance for sports participants should be emphasized.


Neoadjuvant Chemotherapy Plays an Adverse Role in the Prognosis of Grade 2 Breast Cancer.
Background: The role of neoadjuvant chemotherapy (NAC) in the prognosis of breast cancer among patients with grade 2 tumors remains unclear. As such, we aimed to explore the relationships between NAC and survival outcomes among patients with grade 2 breast cancer. Materials and Methods: We collected data on 726 breast cancer patients with grade 2 tumors and at least 5-years of follow-up from the date of diagnosis. We then conducted survival analyses to examine the association between NAC and disease-free survival (DFS) and overall survival (OS). The role of NAC in prognosis was further examined in subgroup analyses, with patients stratified according to molecular subtypes, histological grade, ER status, PR status, HER2 status and Ki67 index. We also determined the main sites of local recurrence, as well as these organs involved in distant metastasis among patients receiving NAC. Finally, we analyzed independent predictive factors for DFS and OS using Cox regression analyses. Results: Among patients who received NAC, the prevalence of pathologic complete response (pCR) was 9.87% (23/233), with 32.6% of patients (76/233) experiencing partial response. Survival analyses demonstrated that NAC had an overall adverse effect on DFS and OS. Subgroup analyses showed that patients who received NAC had shorter DFS in all molecular subgroups of breast cancer, with exception of triple negative breast cancer (TNBC) patients. NAC was also associated with shorter OS among patients with histological grade of 2 and a low Ki67 index. The main recurrence site was the chest well, while distant metastasis occurred in the bone, liver and lung. In Cox regression analyses, we found that NAC was an independent predictor for DFS, but not for OS. Conclusions: NAC may have an adverse effect on breast cancer prognosis among patients with grade 2 tumors. These patients need not receive NAC, except when the patient has a strong desire for breast conservation, and this is unlikely to be achieved in the absence of NAC.


Low rates of child testing for HIV persist in a high-risk area of East Africa.
Children in low- and middle-income countries (LMIC) are the least touched by recent successes in the diagnosis and treatment of HIV/AIDS globally. Early treatment is essential for a child's longer and higher quality of life; however, by 2011, only a small proportion of HIV-seropositive children in LMIC countries were receiving treatment, in part because of persisting low rates of diagnosis. This study of the prevalence and characteristics of children tested for HIV was embedded in the Coping with HIV/AIDS in Tanzania (CHAT) study in which HIV-seropositive and HIV-seronegative adults, and adults with unknown HIV status were asked about HIV testing for their children. Data were gathered from November 2009 to August 2010 during the scale-up of Prevention of Mother To Child Transmission and Early Infant Diagnosis programs in the region. Reports on 1776 children indicate that 31.7% of all children were reported to have been tested, including only 42.9% of children with an HIV-seropositive caregiver. In general, children more likely to be HIV tested were biological children of study participants, younger, of widowed adults, living in urban areas, and of HIV-seropositive parents/caregivers. Children belonging to the two indigenous tribes, Chagga and Pare, were more likely to be tested than those from other tribes. Rates of testing among children less than two years old were low, even for the HIV-seropositive caregiver group. The persistence of low testing rates is discussed in terms of the accessibility and acceptability of child testing in resource poor settings.


Age dependent increase in the levels of osteopontin inhibits skeletal muscle regeneration.
Skeletal muscle regeneration following injury is accompanied by rapid infiltration of macrophages, which play a positive role in muscle repair. Increased chronic inflammation inhibits the regeneration of dystrophic muscle, but the properties of inflammatory cells are not well understood in the context of normal muscle aging. This work uncovers pronounced age-specific changes in the expression of osteopontin (OPN) in CD11b+ macrophages present in the injured old muscle as well as in the blood serum of old injured mice and in the basement membrane surrounding old injured muscle fibers. Furthermore, young CD11b+ macrophages enhance regenerative capacity of old muscle stem cells even when old myofibers and old sera are present; and neutralization of OPN similarly rejuvenates the myogenic responses of old satellite cells in vitro and notably, in vivo. This study highlights potential mechanisms by which age related inflammatory responses become counter-productive for muscle regeneration and suggests new strategies for enhancing muscle repair in the old.


The bile acid membrane receptor TGR5: a valuable metabolic target.
Bile acids (BAs) are amphipathic molecules that facilitate the uptake of lipids, and their levels fluctuate in the intestines as well as in the circulation depending on food intake. Besides their role in dietary lipid absorption, BAs function as signaling molecules that activate specific BA receptors and trigger downstream signaling cascades. The BA receptors and the signaling pathways they control are not only important in the regulation of BA synthesis and their metabolism, but they also regulate glucose homeostasis, lipid metabolism and energy expenditure - processes relevant in the context of the metabolic syndrome. In addition to the function of the nuclear receptor FXRα in regulating local effects of BAs in the organs of the enterohepatic axis, increasing evidence points to a crucial role of the G-protein-coupled receptor TGR5 in mediating systemic actions of BAs. Here we review the current knowledge on BA receptors, with a strong focus on the cell membrane receptor TGR5, which has emerged as a promising target for intervention in metabolic diseases.


Reducing falls among people living with dementia: A systematic review.
People diagnosed with dementia are at greater risk of falls. Given the increasing incidence of dementia globally, high quality and methodologically rigorous research is needed to inform evidence-based practice initiatives. To describe the published literature related to describing, reducing or preventing fall incidences for people living with dementia including: (1) trends in the total number of intervention and non-intervention studies between 1997 and 2016; (2) the methodological quality of identified intervention studies; and (3) the effectiveness of interventions designed to reduce the incidence of falls among people living with dementia. A systematic review of five databases was conducted to identify studies reporting any aspect of falls incidence for people with a diagnosis of dementia. Studies meeting the eligibility criteria were coded as intervention or non-intervention studies. Intervention studies were assessed using Cochrane's Effective Practice and Organisation of Care tool. Data about the effectiveness of interventions meeting Effective Practice and Organisation of Care criteria were extracted. Seventy-two eligible studies were identified; 57 were non-intervention studies, and 15 were intervention studies. The number of published studies increased between 1997 and 2016, peaking in 2013 (n = 10). Of the 15 intervention studies, seven studies met Effective Practice and Organisation of Care design criteria with one study rated low risk on all eight Effective Practice and Organisation of Care risk of bias domains. One high-risk exercise-based intervention study demonstrated a significant reduction in falls among people living with dementia. There is currently insufficient evidence to endorse any intervention to reduce falls for people living with dementia in any setting. More high-quality intervention studies are needed.


Autophagy in Dictyostelium: Mechanisms, regulation and disease in a simple biomedical model.
Autophagy is a fast-moving field with an enormous impact on human health and disease. Understanding the complexity of the mechanism and regulation of this process often benefits from the use of simple experimental models such as the social amoeba Dictyostelium discoideum. Since the publication of the first review describing the potential of D. discoideum in autophagy, significant advances have been made that demonstrate both the experimental advantages and interest in using this model. Since our previous review, research in D. discoideum has shed light on the mechanisms that regulate autophagosome formation and contributed significantly to the study of autophagy-related pathologies. Here, we review these advances, as well as the current techniques to monitor autophagy in D. discoideum. The comprehensive bioinformatics search of autophagic proteins that was a substantial part of the previous review has not been revisited here except for those aspects that challenged previous predictions such as the composition of the Atg1 complex. In recent years our understanding of, and ability to investigate, autophagy in D. discoideum has evolved significantly and will surely enable and accelerate future research using this model.


A prospective study of the effect of Neospora caninum and BVDV infections on bovine abortions in a dairy herd in Arequipa, Peru.
We used a prospective seroepidemiological approach to investigate endemic abortion in a dairy herd in Arequipa, Peru, and its association with Neospora caninum and bovine viral-diarrhoea virus (BVDV) infections. Between January 2002 and March 2004, 1094 pregnancies were confirmed in 538 cows. Of these, 137 pregnancies (13%) in 121 cows ended in abortion. The serological status to N. caninum was assessed using a single serological screening, whereas BVDV status was assessed at the herd level through consecutive samplings of young stock. Cox proportional-hazards models were used to estimate the effect of N. caninum and BVDV on the hazard of early (between day 42 and day 100 in gestation), and late (after day 100) abortions, respectively. Serological status to N. caninum was included as a dichotomous variable, and the effect of BVDV estimated at the herd level, as a time-dependent seasonal effect. Because data from repeated pregnancies were included, we considered possible lack of independence between observations and included frailty effects into the models. Our models also considered the possible confounding by parity and animal origin. Only multiparity was associated with the hazard of early abortion (HR=2.8 compared to nulliparous heifers). N. caninum seropositivity significantly affected the hazard of late abortion, but interacted with parity. The HRs for Neospora-positive animals were 6.4, 3.7 and 1.9, respectively, for nulliparous heifers, first-lactation cows and multiparous cows. Evidence of BVDV circulating (or not) among the young stock was not associated with abortions, but few cows in this herd were susceptible to incident infection.


Enhanced rare-region effects in the contact process with long-range correlated disorder.
We investigate the nonequilibrium phase transition in the disordered contact process in the presence of long-range spatial disorder correlations. These correlations greatly increase the probability for finding rare regions that are locally in the active phase while the bulk system is still in the inactive phase. Specifically, if the correlations decay as a power of the distance, the rare-region probability is a stretched exponential of the rare-region size rather than a simple exponential as is the case for uncorrelated disorder. As a result, the Griffiths singularities are enhanced and take a non-power-law form. The critical point itself is of infinite-randomness type but with critical exponent values that differ from the uncorrelated case. We report large-scale Monte Carlo simulations that verify and illustrate our theory. We also discuss generalizations to higher dimensions and applications to other systems such as the random transverse-field Ising model, itinerant magnets, and the superconductor-metal transition.


Aortic Valve Stenosis: Diagnostic Approaches and Recommendations of the 2021 ESC/EACTS Guidelines for the Management of Valvular Heart Disease -A Review of the Literature.
Aortic stenosis (AS) is the most common valvular heart disease in Europe and North America requiring a surgical or interventional treatment. Due to demographic changes with an aging population the burden of valvular heart diseases and especially the importance of aortic stenosis (AS) will be growing in future. As the onset of symptoms is associated with a decrease in life expectancy appropriate and early diagnosis are of utmost importance. However, insights of clinical practice underline diagnostic challenges which may lead to a delayed initiation of treatment with an adverse effect on the prognosis. The aim of this review is to display different diagnostic approaches that may be helpful in detecting patients with aortic valve stenosis. This review will focus on both non-invasive and invasive diagnostic approaches that can be implemented in clinical routine. Further-more, we will especially highlight recommendations of the 2021 European guidelines for the management of valvular heart disease. For this review a selective literature research on the databases PubMed and Google Scholar was conducted. Original articles, reviews and meta-analyses were included when meeting our search criteria. Following terms were used in different combinations: Aortic valve stenosis; Aortic stenosis; diagnosis of aortic stenosis; ESC Guidelines for the management of valvular heart disease.


Measurement of whole blood thrombus formation using parallel-plate flow chambers - a practical guide.
Custom-made and commercial parallel-plate flow chambers are widely used for studies of platelet activation and thrombus formation in whole blood at defined shear rates. When used in a reproducible way, such flow chamber devices give valuable information on the thrombogenic potential of human, mouse, or rat blood. This article aims to provide a practical guide for the use of parallel-plate flow chambers in combination with routine microscopic imaging techniques. The following methodological aspects are addressed: preparation of surface coatings, calculation of blood flow and shear rate, control of pre-analytical variables, protocols for routine performing of flow chamber tests with non-coagulating or coagulating blood, and procedures for real-time and end-point analysis of thrombus formation. Frequently encountered experimental problems and artifacts are discussed, as well as possibilities for using flow chamber devices as a diagnostic tool to test antithrombotic medication.


Bioinformatic insights to the ESAG5 and GRESAG5 gene families in kinetoplastid parasites.
Trypanosoma brucei, the causative agent of African sleeping sickness, evades the immune response by expressing a coat of variant surface glycoprotein (VSG). VSG is expressed from a single telomeric expression site (ES), along with a number of expression site associated genes (ESAGs). Thus far, the function of most ESAGs is unknown. One ES contains the serum resistance associated gene (SRA), which confers resistance to trypanosome lytic factor in T. b. rhodesiense. Only three other ESAGs -5, 6 and 7 - are present in this ES. ESAGs 6 and 7 encode a heterodimeric transferrin receptor, but the function of ESAG5 has not been identified. We present here a bioinformatic analysis of ESAG5 and distinguish between T. brucei-specific ESAGs and Genes Related to ESAG5 (GRESAGs), which occur outside of ESs in chromosomal-internal contexts. Further, a genome-wide survey of these genes across kinetoplastids identifies a family of GRESAG5s in a number of species. Analysis of phylogenetic relationships indicates that this family may have evolved from a single ancestral copy. Predicted properties of (GR)ESAG5 proteins indicate a glycosylated protein containing either a signal peptide or transmembrane domain. Further analysis indicates a possible relationship to the lipid transfer/lipopolysaccharide-binding family which includes the bactericidal/permeability increasing (BPI) protein. Together, these results provide insights into the structure and evolution of an important extended gene family, and present a number of testable hypotheses which will aid in elucidating the function of ESAG5.


Bacterial vaginosis, Atopobium vaginae and nifuratel.
As bacterial vaginosis (BV) is a potential cause of obstetric complications and gynecological disorders, there is substantial interest in establishing the most effective treatment. Standard treatment - metronidazole or clindamycin, by either vaginal or oral route � is followed by relapses in about 30% of cases, within a month from treatment completion. This inability to prevent recurrences reflects our lack of knowledge on the origins of BV. Atopobium vaginae has been recently reported to be associated with BV in around 80% of the cases and might be involved in the therapeutic failures. This review looks at the potential benefits of nifuratel against A. vaginae compared to the standard treatments with metronidazole and clindamycin. In vitro, nifuratel is able to inhibit the growth of A. vaginae, with a MIC range of 0.125-1 μg/mL; it is active against G. vaginalis and does not affect lactobacilli. Metronidazole is active against A. vaginae only at very high concentrations (8-256 μg/mL); it is partially active against G. vaginalis and also has no effect on lactobacilli. Clindamycin acts against A. vaginae with an MIC lower than 0.125 μg/mL and is active on G. vaginalis but it also affects lactobacilli, altering the vaginal environment. These observations suggest that nifuratel is probably the most valid therapeutic agent for BV treatment.


Health-Related Quality of Life Following Allogeneic Hematopoietic Cell Transplantation with Omidubicel versus Umbilical Cord Blood.
Omidubicel is an advanced cell therapy derived from umbilical cord blood (UCB) for use in allogeneic hematopoietic cell transplantation (HCT). A recent randomized phase 3 clinical trial demonstrated faster engraftment, shorter length of hospital stays, and lower rates of infection with omidubicel compared with standard UCB transplantation in patients with high-risk hematologic malignancies. Despite the proven clinical benefits of omidubicel, its impact on health-related quality of life (HRQL) from the patient's perspective has not been described. This study analyzed patient-reported HRQL measures collected prospectively in the randomized phase 3 trial comparing omidubicel to standard UCB transplantation. A total of 108 patients at 33 international stem cell transplantation centers underwent myeloablative allogeneic HCT with either omidubicel or standard UCB. Patients completed serial HRQL questionnaires at screening and on days 42, 100, 180, and 365 post-transplantation. The HRQL surveys included the Functional Assessment of Cancer Therapy-Bone Marrow Transplant (FACT-BMT), a 50-item cancer-specific questionnaire assessing physical, functional, emotional, social/family, and HCT-specific well-being, and the EuroQol 5-Dimension 3-Level, a 5-item generic HRQL survey. A mixed model with repeated measures was used to compare changes in HRQL from baseline in the 2 treatment arms. The average change in HRQL scores over time was compared by estimating the difference in the area under the curve (AUC) in each treatment group. Seventy-five patients (omidubicel arm, n = 37; standard UCB arm, n = 38) who completed the FACT-BMT at baseline and on 1 or more follow-up visits were included in this study. Baseline characteristics were similar in the 2 treatment arms. Over the first year post-transplantation, the AUCs of mean changes in physical, functional, and total FACT-BMT scores indicated significantly better HRQL with omidubicel (P < .05), with mean differences across time points ranging from 1.4 to 3.1 points, 1.6 to 3.2 points, and 7.2 to 11.0 points, respectively. The minimal clinically important difference was exceeded at 1 or more time points for each of these measures. The HRQL improvements with omidubicel were observed as early as 42 days post-transplantation and persisted at 1 year, indicating the potential long-term benefits of omidubicel on HRQL. Across all patients, adverse clinical outcomes, such as grade 3 viral infections and lower rates of neutrophil engraftment, were associated with worse HRQL scores. The observed improvements in HRQL measures may reflect the known clinical benefits of omidubicel. Compared with standard UCB, allogeneic HCT with omidubicel resulted in significant and clinically meaningful improvements in patient-reported HRQL measures.