Genetic testing for Duchenne/Becker muscular dystrophy in Johannesburg, South Africa.
Genetic testing for Duchenne/Becker muscular dystrophy (DMD/BMD) mutations initially involved multiplex polymerase chain reaction (mPCR), which targeted two mutation hotspots in the gene and detected deletions in affected males. A newer technology, multiplex ligation-dependent probe amplification (MLPA), was introduced for diagnostic testing in 2007. To evaluate MLPA relative to mPCR as a technique for DMD/BMD diagnostic testing and to establish whether the mutation profile in affected individuals differs between different South African ethnic groups. METHODS; From January 2000 - May 2007, genetic diagnostic testing for DMD/BMD was undertaken in 128 male patients using mPCR. From May 2007 onwards, MLPA replaced this technique and 261 males were investigated. MLPA is a kit-based technology available from MRC-Holland.Results. Of the 128 and 261 probands tested using mPCR and MLPA, respectively, 31% and 34% were found to carry a deletion mutation. Further, MLPA could detect duplication mutations (11.5%), complex rearrangements (1.5%) and small mutations (1.5%). In black patients, deletion mutations were found to cluster in the 3' region of the gene. No population-specific pathogenic mutations were found. The mutation detection rate for mPCR and MLPA is similar for deletion mutations, but MLPA proved to be a better diagnostic approach as it could detect other types of mutations as well, including duplications, complex rearrangements and small mutations. MLPA could also diagnose mutation status in at-risk female relatives, which is not possible with mPCR.


Twitching in sensorimotor development from sleeping rats to robots.
It is still not known how the 'rudimentary' movements of fetuses and infants are transformed into the coordinated, flexible and adaptive movements of adults. In addressing this important issue, we consider a behavior that has been perennially viewed as a functionless by-product of a dreaming brain: the jerky limb movements called myoclonic twitches. Recent work has identified the neural mechanisms that produce twitching as well as those that convey sensory feedback from twitching limbs to the spinal cord and brain. In turn, these mechanistic insights have helped inspire new ideas about the functional roles that twitching might play in the self-organization of spinal and supraspinal sensorimotor circuits. Striking support for these ideas is coming from the field of developmental robotics: when twitches are mimicked in robot models of the musculoskeletal system, the basic neural circuitry undergoes self-organization. Mutually inspired biological and synthetic approaches promise not only to produce better robots, but also to solve fundamental problems concerning the developmental origins of sensorimotor maps in the spinal cord and brain.


Effects of systemic inhibition of neuronal nitric oxide synthase in diabetic rats.
Diabetes is associated with alterations in nitric oxide-mediated vasomotor function. The role of nitric oxide generated via the neuronal nitric oxide synthase pathway in the control of systemic and renal hemodynamics in diabetes has not been studied. To explore the hypothesis that diabetic vascular dysfunction is in part caused by altered neuronal nitric oxide synthase activity, systemic and renal hemodynamics were assessed before and after acute inhibition of this enzyme with a specific inhibitor, S-methyl-L-thiocitrulline, in control and diabetic rats. The interaction of this pathway and the renin-angiotensin system was studied in separate groups of rats pretreated with the angiotensin II receptor blocker losartan; these rats were compared with rats treated with losartan alone. Diabetic animals demonstrated higher baseline glomerular filtration rates and filtration fractions. At a low dose, the neuronal nitric oxide synthase inhibitor induced similar dose-dependent pressor responses in control and diabetic rats. Losartan abolished the pressor response in both groups. No changes in renal plasma flow or renal vascular resistance occurred in control rats. In contrast, diabetic rats responded with significant renal vasoconstriction. At a high dose, the renal vasoconstriction was similar in both groups and was not affected by losartan. In conclusion, neuronal nitric oxide synthase-derived nitric oxide plays a role in the control of systemic and renal hemodynamics in normal and diabetic rats. Diabetic rats are more sensitive to the inhibitor, suggesting increased activity of this pathway in the diabetic kidney. Furthermore, renal responses in diabetic rats were attenuated by angiotensin II receptor blockade, whereas losartan alone induced hemodynamic changes that were opposite those seen with neuronal nitric oxide synthase inhibition. This observation implicates angiotensin II as an important modulator of this nitric oxide pathway in diabetes.


Utility of whole-exome sequencing for those near the end of the diagnostic odyssey: time to address gaps in care.
An accurate diagnosis is an integral component of patient care for children with rare genetic disease. Recent advances in sequencing, in particular whole-exome sequencing (WES), are identifying the genetic basis of disease for 25-40% of patients. The diagnostic rate is probably influenced by when in the diagnostic process WES is used. The Finding Of Rare Disease GEnes (FORGE) Canada project was a nation-wide effort to identify mutations for childhood-onset disorders using WES. Most children enrolled in the FORGE project were toward the end of the diagnostic odyssey. The two primary outcomes of FORGE were novel gene discovery and the identification of mutations in genes known to cause disease. In the latter instance, WES identified mutations in known disease genes for 105 of 362 families studied (29%), thereby informing the impact of WES in the setting of the diagnostic odyssey. Our analysis of this dataset showed that these known disease genes were not identified prior to WES enrollment for two key reasons: genetic heterogeneity associated with a clinical diagnosis and atypical presentation of known, clinically recognized diseases. What is becoming increasingly clear is that WES will be paradigm altering for patients and families with rare genetic diseases.


Chondrosarcoma in the humerus of a goat.
The clinical, radiographical and histo-pathological findings of a chondrosarcoma in the humerus of a goat are discussed. An 11-year-old female Saanen goat was admitted for evaluation of chronic, progressive left forelimb lameness. Clinical examination revealed signs of pain in the shoulder region. Radiographs of the scapulohumeral joint showed a large lytic lesion of the proximal third of the humerus with cortical destruction, intralesional calcification and marked periosteal reactions. Differential diagnoses included aggressive bone lesions as seen with osteomyelitis or a primary bone tumour, such as osteosarcoma, chondrosarcoma or fibrosarcoma. The goat was euthanatized for humane reasons because of the poor prognosis. On the basis of the histopathological findings, the lesion was diagnosed as grade II chondrosarcoma. The tumour had already spread to the lungs and the left prescapular lymph node at the time of diagnosis.


[Study of non-participation in the breast cancer screening program in the city of Valencia (Spain)].
To determine the reasons for non-participation of women in a breast cancer screening program. We performed an observational, cross-sectional study in women who were invited to participate in the breast cancer screening program in the city of Valencia and who failed to attend. The women were interviewed in their homes through a questionnaire consisting of 25 questions grouped as follows: knowledge of the program, reasons for nonparticipation, attitudes or beliefs and characteristics of the interviewees (socioeconomic and educational variables and age). A descriptive analysis was performed. Differences in the characteristics of women were evaluated through bivariate analysis. Multivariate analysis was performed through logistic regression. A total of 783 addresses were used, 411 questionnaires were completed and 361 were analyzed. The 50 questionnaires used in the pilot study were excluded because, as a consequence of this study, the questionnaire had been modified. Most of the women (93.4%) (CI: 90.3-95.7) remembered having an appointment. The main reason for not attending was being screened in another health service in 48.8% (CI: 43.6-53.9), followed by various personal reasons in 16.1% (CI: 12.3-19.9) and inability to keep the appointment at the specified time in 15.5% (CI: 11.8-19.2). Comparison of women in the middle and upper social classes with those in the lower classes revealed important differences. Middle and upper class women had a higher probability of knowing about the program and of being on hormone replacement therapy (HRT) and the most frequent reason for non-attendance was attendance at another program run by another health service. In lower class women the reasons for non-attendance were fear, not believing the program to be important to health and inability to keep the appointment at the specified time. In the multivariate analysis, the variables that entered the model were HRT, social class and education. Thus, middle or upper class women undergoing HRT and with secondary or higher education had a higher probability of being screened. The profile of non-participating women belongs on the one hand to those in the middle or upper social classes, undergoing HRT and being screened by other health services and, on the other, to a group of women of low social class, whose reason for non-participation is fear and timetable difficulties.


Sociodemographic moderators of relations of neighborhood safety to physical activity.
This study aimed to investigate gender, race/ethnicity, education, and income as moderators of relations of perceived neighborhood crime, pedestrian, and traffic safety to physical activity. Participants were from two samples: adults (N = 2199, age = 25-65 yr) and older adults (N = 718, age = 66+ yr) from high- and low-walkable neighborhoods in the Washington, DC, and Seattle, Washington, areas. Neighborhood safety and transportation and leisure walking were assessed via survey, and moderate to vigorous physical activity was assessed using accelerometers. Sociodemographic moderators were investigated using interaction terms and follow-up within-group tests from mixed-effects regression models. Overall direct effects of safety on physical activity were not found, with one exception. Seven interactions were found in each sample. Interactions were found for all physical activity outcomes, although total moderate to vigorous physical activity was involved in more interactions in adults than older adults. Half of the interactions revealed significant positive relations of pedestrian and traffic safety to physical activity in the more affluent/advantaged group (i.e., high education, high income, and non-Hispanic white) and null associations in the less affluent/advantaged group. Race/ethnicity was a moderator only in older adults. One-third of the interactions involved gender; half of these involved crime safety. Interactions involving crime safety showed nonsignificant positive trends in the more affluent/advantaged group and women and nonsignificant negative trends in the less affluent/advantaged group and men. Sociodemographic moderators of neighborhood safety explained some of the variation in adults' and older adults' physical activity. Patterns suggested positive associations between safety and physical activity in participants with more affluent/advantaged sociodemographic characteristics, although some patterns were inconsistent, particularly for gender. More refined conceptualizations and measures of safety are needed to understand if and how these constructs influence physical activity.


Tyrannosaur ageing.
Rate of ageing in tyrannosaurs was calculated from parameters of Weibull functions fitted to survival curves based on the estimated ages at death of fossilized remains. Although tyrannosaurs are more closely related to birds than to mammals, they apparently aged at rates similar to mammals of comparable size. Rate of growth in body mass of tyrannosaurs was similar to that of large mammals, and their rates of ageing were consistent with the estimated extrinsic mortality, which is strongly correlated with the rate of ageing across birds and mammals. Thus, tyrannosaurs appear to have had life histories resembling present-day large terrestrial mammals. Rate of ageing in warm-blooded vertebrates appears to be adjusted in response to extrinsic mortality and potential lifespan, independently of both physiological and developmental rates. However, individuals in species with the slowest rates of ageing suffer the highest proportion of ageing-related mortality, hence potentially strong selection to further postpone senescence. Thus, the longest observed lifespans in birds, tyrannosaurs and mammals might be close to the maximum possible.


Impact of obesity on critical care treatment in adult patients.
Obese patients seem to have worse outcomes and more complications during intensive care unit (ICU) stay. This study describes the clinical course, complications and prognostic factors of obese patients admitted to an intensive care unit compared to a control group of nonobese patients. Retrospective observational study. A 10-bed adult intensive care unit in a university-affiliated hospital. All patients admitted to the intensive care unit over 52 months (April 01/2005 to November 30/2008) were included. Obese patients were defined as those with a body mass index (BMI) ≥ 30 Kg/M2. Demographic and intensive care unit related data were also collected. An clinical and demographical matching group of eutrophic patients selected from the data base as comparator for mortality and morbidity outcomes. The Mann-Whitney test was used for numeric data comparisons and the Chi Square test for categorical data comparisons. Two hundred nineteen patients were included. The obese group (n=73) was compared to the eutrophic group (n= 146). Most of this group BMI ranged between 30 - 35 Kg/M2. Only ten patients had body mass index ≥40 Kg/M2. Significant differences between the obese and eutrophic groups were observed in median APACHE II score (16 versus 12, respectively; p<0.05) and median intensive care unit length of stay (7 versus 5 days respectively; p<0,05). No significant differences were seen regarding risk of death, mortality rate, mechanical ventilation needs, days free of mechanical ventilation and tracheostomy rates. The observed mortality was higher than the APACHE II-predicted for both groups, but the larger differences were seen for morbid obese patients (BMI ≥40 Kg/M2). Obesity did not increase the mortality rate, but improved intensive care unit length of stay. The current prognostic scoring systems do not include BMI, possibly underestimating the risk of death, and other quality of care indexes in obese patients. New studies could be useful to clarify how body mass index impacts the mortality rate.


Validity of sentinel lymph node biopsy in Taiwanese breast cancer patients.
The clinical validity of sentinel lymph node (SLN) biopsy in Taiwanese breast cancer patients from 16 institutes in two consecutive periods was analyzed. Data from the initial period between January 3, 1999 and July 2004 and data from the later period after August 2004 to February 2005 were compared. Data on the use of a tracer, SLN identification, harvest and examination, false negative rate (FNR) and the number of patients spared axillary lymph node dissection (ALND) were analyzed. A total of 3308 patients with clinical axilla negative breast cancer underwent SLN biopsies in 16 hospitals (11 in both periods and 5 in the period before August 2004). Comparison of data from the two periods revealed that in the later period, use of combined blue dye and isotope tracer increased from 40% to 80%, the 95% SLN biopsy success rate increased from 50% to 80%, hospitals conducting intraoperative SLN examination increased from 80% to 93%, and the 95% match to permanent section rate increased from 30% to 80%. In the initial period, eight hospitals had less than 7% FNR, while in the later period this had decreased to 4.1% in these hospitals. Another three hospitals had FNR greater than 7% in the initial period, which had decreased to 0% in the later period. Hospitals with more than 100 cases of SLN biopsy had an average 3.8% FNR, whereas hospitals with less than 100 cases of SLN biopsy experience had an 8.2% FNR (p < 0.01). This study found that the SLN biopsy success rate increased after the use of combined tracers and with experience. FNR was controlled to within 5% among breast surgeons with accumulated experience exceeding 100 cases. It is recommended that ALND-sparing surgery be suspended temporarily in hospitals with FNR greater than 5%.


Complexity in pulsed nonlinear laser systems interrogated by permutation entropy.
Permutation entropy (PE) has a growing significance as a relative measure of complexity in nonlinear systems. It has been applied successfully to measuring complexity in nonlinear laser systems. Here, PE and weighted permutation entropy (WPE) are discovered to show an unexpected inversion to higher values, when characterizing the complexity at the characteristic frequencies of nonlinear drivers in laser systems, for output power sequences which are pulsed. The cause of this inversion is explained and its presence can be used to identify when irregular dynamics transform into a fairly regular pulsed signal (with amplitude and timing jitter). When WPE is calculated from experimental output power time series from various nonlinear laser systems as a function of delay time, both the minimum value of WPE, and the width of the peak in the WPE plot are shown to be indicative of the level of amplitude variation and timing jitter present in the pulsed output. Links are made with analysis using simulated time series data with systematic variation in timing jitter and/or amplitude variations.


Serial exercise testing in children, adolescents and young adults with Senning repair for transposition of the great arteries.
Patients with Senning repair for complete transposition of the great arteries (d-TGA) show an impaired exercise tolerance. Our aim was to investigate changes in exercise capacity in children, adolescents and adults with Senning operation. Peak oxygen uptake (peak VO2), oxygen pulse and heart rate were assessed by cardiopulmonary exercise tests (CPET) and compared to normal values. Rates of change were calculated by linear regression analysis. Right ventricular (RV) function was assessed by echocardiography. Thirty-four patients (22 male) performed 3.5 (range 3-6) CPET with an interval of ≥ 6 months. Mean age at first assessment was 16.4 ± 4.27 years. Follow-up period averaged 6.8 ± 2 years. Exercise capacity was reduced (p<0.0005) and the decline of peak VO2 (-1.3 ± 3.7 %/year; p=0.015) and peak oxygen pulse (-1.4 ± 3.0 %/year; p=0.011) was larger than normal, especially before adulthood and in female patients (p<0.01). During adulthood, RV contractility changes were significantly correlated with the decline of peak oxygen pulse (r= -0.504; p=0.047). In patients with Senning operation for d-TGA, peak VO2 and peak oxygen pulse decrease faster with age compared to healthy controls. This decline is most obvious during childhood and adolescence, and suggests the inability to increase stroke volume to the same extent as healthy peers during growth. Peak VO2 and peak oxygen pulse remain relatively stable during early adulthood. However, when RV contractility decreases, a faster decline in peak oxygen pulse is observed.


Predictive models of molecular machines involved in Caenorhabditis elegans early embryogenesis.
Although numerous fundamental aspects of development have been uncovered through the study of individual genes and proteins, system-level models are still missing for most developmental processes. The first two cell divisions of Caenorhabditis elegans embryogenesis constitute an ideal test bed for a system-level approach. Early embryogenesis, including processes such as cell division and establishment of cellular polarity, is readily amenable to large-scale functional analysis. A first step toward a system-level understanding is to provide 'first-draft' models both of the molecular assemblies involved and of the functional connections between them. Here we show that such models can be derived from an integrated gene/protein network generated from three different types of functional relationship: protein interaction, expression profiling similarity and phenotypic profiling similarity, as estimated from detailed early embryonic RNA interference phenotypes systematically recorded for hundreds of early embryogenesis genes. The topology of the integrated network suggests that C. elegans early embryogenesis is achieved through coordination of a limited set of molecular machines. We assessed the overall predictive value of such molecular machine models by dynamic localization of ten previously uncharacterized proteins within the living embryo.


Queen Quality and the Impact of Honey Bee Diseases on Queen Health: Potential for Interactions between Two Major Threats to Colony Health.
Western honey bees, Apis mellifera, live in highly eusocial colonies that are each typically headed by a single queen. The queen is the sole reproductive female in a healthy colony, and because long-term colony survival depends on her ability to produce a large number of offspring, queen health is essential for colony success. Honey bees have recently been experiencing considerable declines in colony health. Among a number of biotic and abiotic factors known to impact colony health, disease and queen failure are repeatedly reported as important factors underlying colony losses. Surprisingly, there are relatively few studies on the relationship and interaction between honey bee diseases and queen quality. It is critical to understand the negative impacts of pests and pathogens on queen health, how queen problems might enable disease, and how both factors influence colony health. Here, we review the current literature on queen reproductive potential and the impacts of honey bee parasites and pathogens on queens. We conclude by highlighting gaps in our knowledge on the combination of disease and queen failure to provide a perspective and prioritize further research to mitigate disease, improve queen quality, and ensure colony health.


Reasons for urban trail use predict levels of trail-related physical activity.
Efforts to increase community levels of physical activity through the development of multiuse urban trails could be strengthened by information about factors predicting trail use. This study examined whether reasons for trail use predict levels of physical activity on urban trails. Adults (N = 335) living within a 1-mile buffer zone of urban trails in Chicago, Dallas, and Los Angeles completed a self-report measure assessing demographics, reason for trail use, and physical activity on the trail. Accelerometers measured total daily moderate-to-vigorous physical activity (MVPA). Environmental features of the urban trail were assessed with the Systematic Pedestrian and Cyclist Environmental Scan for trails measure. Multivariate regression analyses were conducted that accounted for clustering of individuals within trail segments. After controlling for demographic and environmental factors and total daily MVPA, reasons for trail use significantly predicted recreational but not transportation activity. Recreational trail activity was greater for participants who reported exercise and health reasons for trail use as compared with other reasons (ie, social interaction, enjoying nature, walking pets) for recreational trail use. To increase the use of urban trails, it may be useful to promote the health and exercise benefits of recreational trail use.


Multiple lineages of the mitochondrial gene NADH dehydrogenase subunit 1 (ND1) in parasitic helminths: implications for molecular evolutionary studies of facultatively anaerobic eukaryotes.
The task of using partial ND1 sequences to infer a phylogeny for species of the genus Paragonimus (Trematoda: Digenea) was complicated by the discovery of at least two ND1 lineages within individual worms. The divergence of the ND1 lineages is shown by phylogenetic analysis not only to predate the divergence of the three Paragonimus species or species groups investigated but also the divergence of some trematode families. Some sequences are clearly pseudogenes as they contain single base deletions and/or premature termination codons. The presence of both pseudogenes and/or mitochondrial heteroplasmy are invoked to explain the presence of multiple and divergent ND1 lineages in these trematodes, which have two distinct cytochemical types of mitochondria. The implications for phylogenetic studies generally and of parasitic helminths specifically, using ND1 sequence data, are discussed. The ability of these organisms to adapt their metabolic processes to the variable availability of oxygen as an electron acceptor are proposed to explain some of the molecular diversity observed in parasitic helminths and possibly also in other anaerobically adapted eukaryotes.


Impact of obesity-induced type 2 diabetes on long-term outcomes following stroke.
Diabetes is associated with poor recovery profiles following stroke. The pathophysiological mechanisms by which diabetes mediates neurological recovery after stroke are debatable. A recent paper published in the Clinical Science by Pintana et al. (Clinical Science (2019)133, 1367-1386) provides a possible explanation for the underlying mechanisms of poor long-term motor recovery after stroke in obesity-induced diabetes animal model. Authors report that stroke-induced neurogenesis and parvalbumin (PV)+ interneuron-mediated neuroplasticity is severely impaired due to obesity-induced type 2 diabetes (T2D). Poor long-term motor recovery after stroke in comorbid obese and diabetic mice was not associated with stroke-induced grey or white matter damage. Understanding these mechanisms is crucial to develop therapeutic strategies to improve recovery in the obesity-induced diabetic population. The strength of the present study lies in the use of a comorbid obese/diabetic animal model, which is more likely to reflect the clinical scenario. However, these findings should be understood from the context of this specific animal model and whether these findings hold true for another variant of the obesity/T2D model warrants further consideration. This is an interesting study from the perspective of understanding the stroke pathology in T2D; however, the interaction of microvascular changes (including vascular modelling, angiogenesis), oxidative stress and insulin resistance (IR) associated with T2D and poor recovery profile merit further discussions. Given the increasing burden of obesity, diabetes and/or stroke globally, understanding of mechanisms may be useful in developing cardiovascular risk management pathways in this subgroup of population who are at increased risk of poor clinical outcomes following acute stroke.


Eating disorder features in indigenous Aboriginal and Torres Strait Islander Australian peoples.
Obesity and related cardiovascular and metabolic conditions are well recognized problems for Australian Aboriginal and Torres Strait Islander peoples. However, there is a dearth of research on relevant eating disorders (EDs) such as binge eating disorder in these groups. Data were obtained from interviews of 3047 (in 2005) and 3034 (in 2008) adults who were participants in a randomly selected South Australian household survey of individuals' age > 15 years. The interviewed comprised a general health survey in which ED questions were embedded. Data were weighted according to national census results and comprised key features of ED symptoms. In 2005 there were 94 (85 weighted) First Australian respondents, and in 2008 65 (70 weighted). Controlling for secular differences, in 2005 rates of objective binge eating and levels of weight and shape influence on self-evaluation were significantly higher in indigenous compared to non-indigenous participants, but no significant differences were found in ED features in 2008. Whilst results on small numbers must be interpreted with caution, the main finding was consistent over the two samples. For First Australians ED symptoms are at least as frequent as for non-indigenous Australians.


Improved detection of biomarkers in cervico-vaginal mucus (CVM) from postpartum cattle.
In the postpartum cow, early diagnosis of uterine disease is currently problematic due to the lack of reliable, non-invasive diagnostic methods. Cervico-vaginal mucus (CVM) is an easy to collect potentially informative source of biomarkers for the diagnosis and prognosis of uterine disease in cows. Here, we report an improved method for processing CVM from postpartum dairy cows for the measurement of immune biomarkers. CVM samples were collected from the vagina using gloved hand during the first two weeks postpartum and processed with buffer alone or buffer containing different concentrations of the reducing agents recommended in standard protocols: Dithiothriotol (DTT) or N-Acetyl-L-Cysteine (NAC). Total protein was measured using the bicinchoninic acid (BCA) assay; interleukin 6 (IL-6), IL-8 and α1-acid glycoprotein (AGP) were measured by ELISA. We found that use of reducing agents to liquefy CVM affects protein yield and the accuracy of biomarker detection. Our improved protocol results in lower protein yields but improved detection of cytokines and chemokines. Using our modified method to measure AGP in CVM we found raised levels of AGP at seven days postpartum in CVM from cows that went on to develop endometritis. We conclude that processing CVM without reducing agents improves detection of biomarkers that reflect uterine health in cattle. We propose that measurement of AGP in CVM during the first week postpartum may identify cows at risk of developing clinical endometritis.


Results from the Trial Using Motivational Interviewing, Positive Affect, and Self-Affirmation in African Americans with Hypertension (TRIUMPH).
Our objective was to determine the effectiveness of combining positive affect and self-affirmation strategies with motivational interviewing in achieving blood pressure control among hypertensive African Americans (AA) compared with AA hypertensives in an education-only control group. Randomized trial. Ambulatory practices in the South Bronx and Harlem, New York City. African American adults with uncontrolled hypertension. Participants were randomized to a positive affect and self-affirmation intervention or an education control group. The positive affect and self-affirmation intervention involved having participants think about things that made them happy and that reminded them of their core values on a daily basis. These strategies were reinforced every two months through motivational interviewing. The control arm received a workbook of strategies on blood pressure control. All participants were called every two months for one year. Blood pressure control rate. A total of 238 participants were randomized. The average age was 56 ± 11 years, approximately 70% were female, 80% were not married, and up to 70% had completed high school. There was no difference in control rates between the intervention and the control group. However, at one year, female participants were more likely to be controlled. Participants with high depressive symptoms or high perceived stress at baseline were less likely to be controlled. While this study did not demonstrate an intervention effect, it does provide important insight into the psychosocial factors that may underlie blood pressure control in African Americans. Implications for future behavioral intervention trials are discussed.


Effect of octanoic acid-rich formula on plasma ghrelin levels in cachectic patients with chronic respiratory disease.
For cachectic patients with chronic respiratory disease (CRD), conventional enteral nutrition formula is an optional treatment to maintain energy balance. The molecular mechanisms by which enteral nutrition formula controls appetite and weight remain unknown. We examined whether enteral nutrition formula rich in octanoic acids would increase plasma levels of ghrelin, an appetite-stimulating hormone produced in the stomach, in cachectic patients with CRD. Plasma ghrelin profiles in cachectic patients with CRD were assessed and compared with those in age- and sex-matched controls. Plasma levels of acyl-ghrelin, an active ghrelin modified by octanoic acids, and desacyl-ghrelin were measured separately. We examined changes in 24-h plasma ghrelin profiles before and after single administration of the formula. We also evaluated the effects of 2-week administration of the formula on plasma ghrelin levels and nutritional status in patients. The ratio of acyl-ghrelin to desacyl-ghrelin in plasma was lower in patients than in controls. Single administration of the formula did not change plasma desacyl-ghrelin levels, but induced an increase in acyl-ghrelin levels. Two-week treatment with the formula was effective in increasing weight and acyl-ghrelin, along with improving nutritional status in patients. These results show that the formula contributes to increased weight, which may be associated with induction of acyl-ghrelin production in cachectic patients with CRD.


The impact of large terrestrial carnivores on Pleistocene ecosystems.
Large mammalian terrestrial herbivores, such as elephants, have dramatic effects on the ecosystems they inhabit and at high population densities their environmental impacts can be devastating. Pleistocene terrestrial ecosystems included a much greater diversity of megaherbivores (e.g., mammoths, mastodons, giant ground sloths) and thus a greater potential for widespread habitat degradation if population sizes were not limited. Nevertheless, based on modern observations, it is generally believed that populations of megaherbivores (>800 kg) are largely immune to the effects of predation and this perception has been extended into the Pleistocene. However, as shown here, the species richness of big carnivores was greater in the Pleistocene and many of them were significantly larger than their modern counterparts. Fossil evidence suggests that interspecific competition among carnivores was relatively intense and reveals that some individuals specialized in consuming megaherbivores. To estimate the potential impact of Pleistocene large carnivores, we use both historic and modern data on predator-prey body mass relationships to predict size ranges of their typical and maximum prey when hunting as individuals and in groups. These prey size ranges are then compared with estimates of juvenile and subadult proboscidean body sizes derived from extant elephant growth data. Young proboscideans at their most vulnerable age fall within the predicted prey size ranges of many of the Pleistocene carnivores. Predation on juveniles can have a greater impact on megaherbivores because of their long interbirth intervals, and consequently, we argue that Pleistocene carnivores had the capacity to, and likely did, limit megaherbivore population sizes.


Interim analysis of safety and efficacy of ruxolitinib in patients with myelofibrosis and low platelet counts.
Ruxolitinib, a Janus kinase 1 and 2 inhibitor, demonstrated improvements in spleen volume, symptoms, and survival over placebo and best available therapy in intermediate-2 or high-risk myelofibrosis patients with baseline platelet counts ≥100 × 109/L in phase III studies. The most common adverse events were dose-dependent anemia and thrombocytopenia, which were anticipated because thrombopoietin and erythropoietin signal through JAK2. These events were manageable, rarely leading to treatment discontinuation. Because approximately one-quarter of MF patients have platelet counts <100 × 109/L consequent to their disease, ruxolitinib was evaluated in this subset of patients using lower initial doses. Interim results of a phase II study of ruxolitinib in myelofibrosis patients with baseline platelet counts of 50-100 × 109/L are reported. Ruxolitinib was initiated at a dose of 5 mg twice daily (BID), and doses could be increased by 5 mg once daily every 4 weeks to 10 mg BID if platelet counts remained adequate. Additional dosage increases required evidence of suboptimal efficacy. Assessments included measurement of spleen volume by MRI, MF symptoms by MF Symptom Assessment Form v2.0 Total Symptom Score [TSS]), Patient Global Impression of Change (PGIC); EORTC QLQ-C30, and safety/tolerability. By week 24, 62% of patients achieved stable doses ≥10 mg BID. Median reductions in spleen volume and TSS were 24.2% and 43.8%, respectively. Thrombocytopenia necessitating dose reductions and dose interruptions occurred in 12 and 8 patients, respectively, and occurred mainly in patients with baseline platelet counts ≤75 × 109/L. Seven patients experienced platelet count increases ≥15 × 109/L. Mean hemoglobin levels remained stable over the treatment period. Two patients discontinued for adverse events: 1 for grade 4 retroperitoneal hemorrhage secondary to multiple and suspected pre-existing renal artery aneurysms and 1 for grade 4 thrombocytopenia. Results suggest that a low starting dose of ruxolitinib with escalation to 10 mg BID may be appropriate in myelofibrosis patients with low platelet counts.


Fluence rate-dependent intratumor heterogeneity in physiologic and cytotoxic responses to Photofrin photodynamic therapy.
Photodynamic therapy (PDT) can lead to the creation of heterogeneous, response-limiting hypoxia during illumination, which may be controlled in part through illumination fluence rate. In the present report we consider (1) regional differences in hypoxia, vascular response, and cell kill as a function of tumor depth and (2) the role of fluence rate as a mediator of depth-dependent regional intratumor heterogeneity. Intradermal RIF murine tumors were treated with Photofrin PDT using surface illumination at an irradiance of 75 or 38 mW cm(-2). Regional heterogeneity in tumor response was examined through comparison of effects in the surface vs. base of tumors, i.e. along a plane parallel to the skin surface and perpendicular to the incident illumination. 75 mW cm(-2) PDT created significantly greater hypoxia in tumor bases relative to their surfaces. Increased hypoxia in the tumor base could not be attributed to regional differences in Photofrin concentration nor effects of fluence rate distribution on photochemical oxygen consumption, but significant depth-dependent heterogeneity in vascular responses and cytotoxic response were detected. At a lower fluence rate of 38 mW cm(-2), no detectable regional differences in hypoxia or cytotoxic responses were apparent, and heterogeneity in vascular response was significantly less than that during 75 mW cm(-2) PDT. This research suggests that the benefits of low-fluence-rate PDT are mediated in part by a reduction in intratumor heterogeneity in hypoxic, vascular and cytotoxic responses.


Headache rate and cost of care following lumbar puncture at a single tertiary care hospital.
American Academy of Neurology guidelines recommend the use of noncutting needles because of lower rates of headache following lumbar puncture in randomized trials. We sought to determine the rate of headache using cutting needles and the potential cost savings of switching to noncutting needles. We retrospectively reviewed the charts of all patients who had a lumbar puncture in the outpatient neurology clinic at a single institution between January 2004 and December 2005. Outcome data included occurrence of headache, back pain, or epidural hematomas within 2 weeks of the procedure. Costs associated with the use of the current system were compared with the projected costs of switching to a noncutting needle system. A total of 274 patients underwent lumbar puncture (62% women, mean age 53 ± 17 years, average weight 178 ± 43 pounds). Of these, 38 (14%) had a post-lumbar puncture headache. Eight patients (3%) reported back pain. No patients had an epidural hematoma. Twelve patients were admitted for a total of 18 hospital days, mainly for headache. Predictors of headache were younger age and no prior aspirin use. The rate of headache associated with the noncutting needle according to published literature is 4%. The estimated cost savings would have been approximately $20,000 per year (or approximately $73 per person). In this single-institution study, use of a noncutting needle would have potentially been associated with less adverse events and less cost. Further studies are warranted, including the possibility of premedication with aspirin.


Codes for multiplex spectrometry.
A number of binary cyclic coding schemes for multiplex spectrometry are discussed and evaluated in terms of a linear, least mean square, unbiased estimate. The optical realization of such codes in dispersion instruments is briefly discussed. We show that there are many advantages both in the construction of the instrument and in its operation which accrue from cyclic codes.


A genome-wide association study for mastitis resistance in phenotypically well-characterized Holstein dairy cattle using a selective genotyping approach.
A decrease in the incidence of bovine mastitis, the costliest disease in the dairy industry, can be facilitated through genetic marker-assisted selective breeding programs. Identification of genomic variants associated with mastitis resistance is an ongoing endeavor for which genome-wide association studies (GWAS) using high-density arrays provide a valuable tool. We identified single nucleotide polymorphisms (SNPs) in Holstein dairy cattle associated with mastitis resistance in a GWAS by using a high-density SNP array. Mastitis-resistant (15) and mastitis-susceptible (28) phenotypic extremes were identified from 224 lactating dairy cows on commercial dairy farm located in Utah based on multiple criteria of mastitis resistance over an 8-month period. Twenty-seven quantitative trait loci (QTLs) for mastitis resistance were identified based on 117 SNPs suggestive of genome-wide significance for mastitis resistance (p ≤ 1 × 10-4), including 10 novel QTLs. Seventeen QTLs overlapped previously reported QTLs of traits relevant to mastitis, including four QTLs for teat length. One QTL includes the RAS guanyl-releasing protein 1 gene (RASGRP1), a candidate gene for mastitis resistance. This GWAS identifies 117 candidate SNPs and 27 QTLs for mastitis resistance using a selective genotyping approach, including 10 novel QTLs. Based on overlap with previously identified QTLs, teat length appears to be an important trait in mastitis resistance. RASGRP1, overlapped by one QTL, is a candidate gene for mastitis resistance.


Testing physical models of passive membrane permeation.
The biophysical basis of passive membrane permeability is well-understood, but most methods for predicting membrane permeability in the context of drug design are based on statistical relationships that indirectly capture the key physical aspects. Here, we investigate molecular mechanics-based models of passive membrane permeability and evaluate their performance against different types of experimental data, including parallel artificial membrane permeability assays (PAMPA), cell-based assays, in vivo measurements, and other in silico predictions. The experimental data sets we use in these tests are diverse, including peptidomimetics, congeneric series, and diverse FDA approved drugs. The physical models are not specifically trained for any of these data sets; rather, input parameters are based on standard molecular mechanics force fields, such as partial charges, and an implicit solvent model. A systematic approach is taken to analyze the contribution from each component in the physics-based permeability model. A primary factor in determining rates of passive membrane permeation is the conformation-dependent free energy of desolvating the molecule, and this measure alone provides good agreement with experimental permeability measurements in many cases. Other factors that improve agreement with experimental data include deionization and estimates of entropy losses of the ligand and the membrane, which lead to size-dependence of the permeation rate.


Postoperative Expansion is not a Primary Cause of Infection in Immediate Breast Reconstruction with Tissue Expanders.
Perioperative infection is the most common and dreaded complication associated with tissue expander (TE) breast reconstruction. Historically, the expansion period was thought to be the time of greatest hazard to the implant. However, recent institutional observations suggest infectious complications occur prior to expansion. This investigation, therefore, was conducted to determine the timing of infectious complications associated with two-stage TE breast reconstructions. Following IRB approval, a retrospective review of all consecutive two-stage immediate TE breast reconstructions at a single institution from November 2007 to November 2011 was conducted. Reconstructions were then divided into two cohorts: those suffering infectious complications and those that did not. Infectious complications including minor cellulitis, major cellulitis, abscess drainage, and explantation were identified. Various operative and patient variables were evaluated in comparison. Eight hundred ninety immediate two-stage TE breast reconstructions met inclusion criteria. Patients suffering infection were older (55.4 years versus 49.3 years; p < 0.001), and more likely to have therapeutic mastectomy (94% versus 61%; p < 0.0001), the use of acellular dermal matrix (ADM; 72.5% versus 54.9%; p = 0.001), and greater initial TE fill (448.6 mL versus 404.7 mL; p = 0.0078). The average time to developing of infectious symptoms was 29.6 days (range 9-142 days), with 94.6% (n = 87) of infections prior to the start of expansion. Perioperative infections in immediate two-stage TE to implant breast reconstructions are significant and occur mostly prior to the start of expansion. Thus, challenging the conventional wisdom that instrumentation during expander filling as the primary cause of implant infections. Possible etiologic factors include greater age, therapeutic mastectomy versus prophylactic mastectomy, larger initial TE fill, and the use of ADM.


Gastric juice protects against the development of esophageal adenocarcinoma in the rat.
The authors investigate the effects of gastric juice on tumorigenesis in a rat model of esophageal adenocarcinoma. In rats treated with the carcinogen methyl-n-amyl nitrosamine, squamous cancer of the esophagus develops in a time- and dose-dependent manner. When methyl-n-amyl nitrosamine treatment is preceded by an operation to induce reflux of duodenal and gastric juice into the esophagus, there is an increased yield of esophageal tumors, many of which are adenocarcinomas. When only gastric juice refluxes into the esophagus, the tumor yield is less and adenocarcinomas are not found. Two hundred seventy 8-week old Sprague-Dawley rats were studied. Twenty unoperated rats served as controls. The remaining rats underwent the following operations: esophagoduodenostomy with gastric and vagal preservation to induce duodenogastroesophageal reflux (n = 48); esophagoduodenostomy with antrectomy and Billroth 1 reconstruction to produce reflux of duodenogastric juice with the exclusion of the antrum (n = 53); esophagoduodenostomy with proximal gastrectomy to induce hypergastrinemia and reflux of duodenogastric juice with exclusion of the body and forestomach (n = 51); esophagoduodenostomy plus total gastrectomy to produce reflux of duodenal juice alone (n = 50); and esophagoduodenostomy with vagal and gastric preservation but with division of the duodenum just beyond the pylorus and reimplantation into the jejunum, 13 cm distal to the esophagoduodenostomy. This produced reflux of duodenal juice with gastric juice diverted downstream, (n = 48). At 10 weeks of age, all rats were given 4 weekly doses of carcinogen (methyl-n-amyl nitrosamine, 25 mg/kg intraperitoneally), and survivors were killed at 36 weeks of age. The prevalence rate of esophageal adenocarcinoma was 30% in rats with duodenogastroesophageal reflux and 87% in rats with reflux of duodenal juice alone. Fifty-six percent of rats with reflux of duodenogastric juice with exclusion of the antrum and 72% of rats with reflux of duodenogastric juice with the exclusion of the body and forestomach developed adenocarcinoma, showing a progression increase in the prevalence of adenocarcinoma as less gastric juice was permitted to reflux with duodenal juice into the esophagus. In this rat model, the presence of gastric juice in refluxed duodenal juice against the development of esophageal adenocarcinoma. The protective effect appears to be due to acid secretion from the stomach. Continuous profound acid suppression therapy may be detrimental by encouraging esophageal metaplasia and tumorigenesis in patients with duodenogastroesophageal reflux.


Cytoprotective Effect of Ferritin H in Renal Ischemia Reperfusion Injury.
Oxidative stress is a major contributor to kidney injury following ischemia reperfusion. Ferritin, a highly conserved iron-binding protein, is a key protein in the maintenance of cellular iron homeostasis and protection from oxidative stress. Ferritin mitigates oxidant stress by sequestering iron and preventing its participation in reactions that generate reactive oxygen species. Ferritin is composed of two subunit types, ferritin H and ferritin L. Using an in vivo model that enables conditional tissue-specific doxycycline-inducible expression of ferritin H in the mouse kidney, we tested the hypothesis that an increased level of H-rich ferritin is renoprotective in ischemic acute renal failure. Prior to induction of ischemia, doxycycline increased ferritin H in the kidneys of the transgenic mice nearly 6.5-fold. Following reperfusion for 24 hours, induction of neutrophil gelatinous-associated lipocalin (NGAL, a urine marker of renal dysfunction) was reduced in the ferritin H overexpressers compared to controls. Histopathologic examination following ischemia reperfusion revealed that ferritin H overexpression increased intact nuclei in renal tubules, reduced the frequency of tubular profiles with luminal cast materials, and reduced activated caspase-3 in the kidney. In addition, generation of 4-hydroxy 2-nonenal protein adducts, a measurement of oxidant stress, was decreased in ischemia-reperfused kidneys of ferritin H overexpressers. These studies demonstrate that ferritin H can inhibit apoptotic cell death, enhance tubular epithelial viability, and preserve renal function by limiting oxidative stress following ischemia reperfusion injury.


Peh production, flagellum synthesis, and virulence reduced in Erwinia carotovora subsp. carotovora by mutation in a homologue of cytR.
Erwinia carotovora subsp. carotovora is a causal agent of soft-rot diseases in a wide variety of plants. Here, we have isolated a new regulatory factor involved in the virulence of E. carotovora subsp. carotovora by in vivo insertional mutagenesis using a transposon Tn5. The gene was homologous to cytR encoding a transcriptional repressor of nucleoside uptake and catabolism genes in Escherichia coli, Salmonella typhimurium, and Vibrio cholerae. Phenotypic characterization of a nonpolar deletion mutant of the cytR homologue (delta cytR) revealed that the delta cytR mutant produced a reduced level of polygalacturonase (Peh) and lost its motility compared to that in the parental strain. With electron microscopy, the delta cytR mutant was shown to be aflagellate. Furthermore, the expression of fliA and fliC (encoding sigma28 and flagellin, respectively) was also reduced in delta cytR mutant. The virulence of delta cytR mutant was reduced in Chinese cabbage and potato compared to that of the parental strain. These results suggest that the CytR homologue of E. carotovora subsp. carotovora positively controls Peh production and flagellum synthesis and plays an important role in its pathogenicity.


Dynein regulates Kv7.4 channel trafficking from the cell membrane.
The dynein motor protein transports proteins away from the cell membrane along the microtubule network. Recently, we found the microtubule network was important for regulating the membrane abundance of voltage-gated Kv7.4 potassium channels in vascular smooth muscle. Here, we aimed to investigate the influence of dynein on the microtubule-dependent internalization of the Kv7.4 channel. Patch-clamp recordings from HEK293B cells showed Kv7.4 currents were increased after inhibiting dynein function with ciliobrevin D or by coexpressing p50/dynamitin, which specifically interferes with dynein motor function. Mutation of a dynein-binding site in the Kv7.4 C terminus increased the Kv7.4 current and prevented p50 interference. Structured illumination microscopy, proximity ligation assays, and coimmunoprecipitation showed colocalization of Kv7.4 and dynein in mesenteric artery myocytes. Ciliobrevin D enhanced mesenteric artery relaxation to activators of Kv7.2-Kv7.5 channels and increased membrane abundance of Kv7.4 protein in isolated smooth muscle cells and HEK293B cells. Ciliobrevin D failed to enhance the negligible S-1-mediated relaxations after morpholino-mediated knockdown of Kv7.4. Mass spectrometry revealed an interaction of dynein with caveolin-1, confirmed using proximity ligation and coimmunoprecipitation assays, which also provided evidence for interaction of caveolin-1 with Kv7.4, confirming that Kv7.4 channels are localized to caveolae in mesenteric artery myocytes. Lastly, cholesterol depletion reduced the interaction of Kv7.4 with caveolin-1 and dynein while increasing the overall membrane expression of Kv7.4, although it attenuated the Kv7.4 current in oocytes and interfered with the action of ciliobrevin D and channel activators in arterial segments. Overall, this study shows that dynein can traffic Kv7.4 channels in vascular smooth muscle in a mechanism dependent on cholesterol-rich caveolae.


Methods to develop and maintain a valid physician registry in evolving information environments.
WHAT DID WE DO?: As the amount of alternative funding for physician services grows in Canada, the usefulness of fee-for-services (FFS) payment data as a source of population-based information declines. This paper describes methods used to develop and validate an anonymous registry of the physician workforce to support policy-relevant analyses in environments where doctors are increasingly funded through diverse arrangements. WHAT DID WE LEARN?: Among the 8,558 physicians in clinical practice in British Columbia in 2004, 97% could be identified via FFS payment data. In 1996, 2000 and 2004, a similar number of physicians in clinical practice (N=340, 326, 290) did not submit FFS claims, and a stable proportion of these doctors (65%, 67%, 69%) could be identified using hospital discharge abstract data. Province-wide, local health areas varied in the proportion of total physicians' expenditures in 2004 attributable to FFS payments (0 to 100%). WHAT ARE THE IMPLICATIONS?: FFS and hospital files could be used in tandem to identify physicians in clinical practice in order to create complete registries intended to support population-based workforce analyses. FFS and alternative funding payment files could be used together to calculate each physician's income and clinical activity, and to identify physicians for whom there is high likelihood of measurement error because they reside in local health areas where a large proportion of clinical activity is not captured. Systems designed to track alternative funding should be uniformly structured within and across jurisdictions to maintain or increase the availability of population-based clinical data useful for secondary analyses. Only then can these data be used to support Canadian policy, management and clinical decision-making.


Diet and the evolution of human amylase gene copy number variation.
Starch consumption is a prominent characteristic of agricultural societies and hunter-gatherers in arid environments. In contrast, rainforest and circum-arctic hunter-gatherers and some pastoralists consume much less starch. This behavioral variation raises the possibility that different selective pressures have acted on amylase, the enzyme responsible for starch hydrolysis. We found that copy number of the salivary amylase gene (AMY1) is correlated positively with salivary amylase protein level and that individuals from populations with high-starch diets have, on average, more AMY1 copies than those with traditionally low-starch diets. Comparisons with other loci in a subset of these populations suggest that the extent of AMY1 copy number differentiation is highly unusual. This example of positive selection on a copy number-variable gene is, to our knowledge, one of the first discovered in the human genome. Higher AMY1 copy numbers and protein levels probably improve the digestion of starchy foods and may buffer against the fitness-reducing effects of intestinal disease.


Who is trustworthy? Predicting trustworthy intentions and behavior.
Existing trust research has disproportionately focused on what makes people more or less trusting, and has largely ignored the question of what makes people more or less trustworthy. In this investigation, we deepen our understanding of trustworthiness. Across six studies using economic games that measure trustworthy behavior and survey items that measure trustworthy intentions, we explore the personality traits that predict trustworthiness. We demonstrate that guilt-proneness predicts trustworthiness better than a variety of other personality measures, and we identify sense of interpersonal responsibility as the underlying mechanism by both measuring it and manipulating it directly. People who are high in guilt-proneness are more likely to be trustworthy than are individuals who are low in guilt-proneness, but they are not universally more generous. We demonstrate that people high in guilt-proneness are more likely to behave in interpersonally sensitive ways when they are more responsible for others' outcomes. We also explore potential interventions to increase trustworthiness. Our findings fill a significant gap in the trust literature by building a foundation for investigating trustworthiness, by identifying a trait predictor of trustworthy intentions and behavior, and by providing practical advice for deciding in whom we should place our trust. (PsycINFO Database Record


Bilevel Optimization for Cost Function Determination in Dynamic Simulation of Human Gait.
Predictive simulation based on dynamic optimization using musculoskeletal models is a powerful approach for studying human gait. Predictive musculoskeletal simulation may be used for a variety of applications from designing assistive devices to testing theories of motor control. However, the underlying cost function for the predictive optimization is unknown and is generally assumed a priori. Alternatively, the underlying cost function can be determined from among a family of possible cost functions, representing an inverse optimal control problem that may be solved using a bilevel optimization approach. In this study, a nested evolutionary approach is proposed to solve the bilevel optimization problem. The lower level optimization is solved by a direct collocation method, and the upper level is solved by a genetic algorithm. We demonstrate our approach to solve different bilevel optimization problems, including finding the weights among three common performance criteria in the cost function for normal human walking. The proposed approach was found to be effective at solving the bilevel optimization problems. This approach should provide practical utility in designing assistive devices to aid mobility, and could yield insights about the control of human walking.


Phosphocitrate inhibits a basic calcium phosphate and calcium pyrophosphate dihydrate crystal-induced mitogen-activated protein kinase cascade signal transduction pathway.
Calcium deposition diseases caused by calcium pyrophosphate dihydrate (CPPD) and basic calcium phosphate (BCP) crystals are a significant source of morbidity in the elderly. We have shown previously that both types of crystals can induce mitogenesis, as well as metalloproteinase synthesis and secretion by fibroblasts and chondrocytes. These responses may promote degradation of articular tissues. We have also shown previously that both CPPD and BCP crystals activate expression of the c-fos and c-jun proto-oncogenes. Phosphocitrate (PC) can specifically block mitogenesis and proto-oncogene expression induced by either BCP or CPPD crystals in 3T3 cells and human fibroblasts, suggesting that PC may be an effective therapy for calcium deposition diseases. To understand how PC inhibits BCP and CPPD-mediated cellular effects, we have investigated the mechanism by which BCP and CPPD transduce signals to the nucleus. Here we demonstrate that BCP and CPPD crystals activate a protein kinase signal transduction pathway involving p42 and p44 mitogen-activated protein (MAP) kinases (ERK 2 and ERK 1). BCP and CPPD also cause phosphorylation of a nuclear transcription factor, cyclic AMP response element-binding protein (CREB), on serine 133, a residue essential for CREB's ability to transactivate. Treatment of cells with PC at concentrations of 10(-3) to 10(-5) M blocked both the activation of p42/p44 MAP kinases, and CREB serine 133 phosphorylation, in a dose-dependent fashion. At 10(-3) M, a PC analogue, n-sulfo-2-aminotricarballylate and citrate also modulate this signal transduction pathway. Inhibition by PC is specific for BCP- and CPPD-mediated signaling, since all three compounds had no effect on serum-induced p42/P44 or interleukin-1beta induced p38 MAP kinase activities. Treatment of cells with an inhibitor of MEK1, an upstream activator of MAPKs, significantly inhibited crystal-induced cell proliferation, suggesting that the MAPK pathway is a significant mediator of crystal-induced signals.


Abdominal abscess. A surgical strategy.
To reassess the role of laparotomy and extraserosal drainage in the treatment of patients with abdominal abscess, we analyzed the course of 79 patients who underwent 97 operations to treat 120 abdominal abscesses during a five-year period. In 66 clinical episodes the abscess was drained by the most direct approach. Sepsis resolved with a single operation In 80% of these patients, five patients (8%) required a second operation for drainage for an abscess, and eight patients (12%) died. In 31 clinical episodes, the abscess was drained by a laparotomy. Sepsis resolved with a single operation in 61% of these patients, seven patients (21%) had a second abscess, six patients (19%) required a second operation to drain a metachronous abscess, and six patients (19%) died. When the location or number of abscesses was diagnosed incorrectly, the success rate of therapy fell substantially. Since most abdominal abscesses can now be accurately diagnosed preoperatively, most abscesses should be drained by a direct approach. Exploratory laparotomy is indicated when preoperative localization is unsuccessful, when sepsis has not resolved after other methods of drainage, or when the patient has a concomitant abdominal condition that must be treated surgically.


Genetically elevated high-density lipoprotein cholesterol through the cholesteryl ester transfer protein gene does not associate with risk of Alzheimer's disease.
There is conflicting evidence whether high-density lipoprotein cholesterol (HDL-C) is a risk factor for Alzheimer's disease (AD) and dementia. Genetic variation in the cholesteryl ester transfer protein (CETP) locus is associated with altered HDL-C. We aimed to assess AD risk by genetically predicted HDL-C. Ten single nucleotide polymorphisms within the CETP locus predicting HDL-C were applied to the International Genomics of Alzheimer's Project (IGAP) exome chip stage 1 results in up 16,097 late onset AD cases and 18,077 cognitively normal elderly controls. We performed instrumental variables analysis using inverse variance weighting, weighted median, and MR-Egger. Based on 10 single nucleotide polymorphisms distinctly predicting HDL-C in the CETP locus, we found that HDL-C was not associated with risk of AD (P > .7). Our study does not support the role of HDL-C on risk of AD through HDL-C altered by CETP. This study does not rule out other mechanisms by which HDL-C affects risk of AD.


Context-dependent variability in the predicted daily energetic costs of disturbance for blue whales.
Assessing the long-term consequences of sub-lethal anthropogenic disturbance on wildlife populations requires integrating data on fine-scale individual behavior and physiology into spatially and temporally broader, population-level inference. A typical behavioral response to disturbance is the cessation of foraging, which can be translated into a common metric of energetic cost. However, this necessitates detailed empirical information on baseline movements, activity budgets, feeding rates and energy intake, as well as the probability of an individual responding to the disturbance-inducing stressor within different exposure contexts. Here, we integrated data from blue whales (Balaenoptera musculus) experimentally exposed to military active sonar signals with fine-scale measurements of baseline behavior over multiple days or weeks obtained from accelerometry loggers, telemetry tracking and prey sampling. Specifically, we developed daily simulations of movement, feeding behavior and exposure to localized sonar events of increasing duration and intensity and predicted the effects of this disturbance source on the daily energy intake of an individual. Activity budgets and movements were highly variable in space and time and among individuals, resulting in large variability in predicted energetic intake and costs. In half of our simulations, an individual's energy intake was unaffected by the simulated source. However, some individuals lost their entire daily energy intake under brief or weak exposure scenarios. Given this large variation, population-level models will have to assess the consequences of the entire distribution of energetic costs, rather than only consider single summary statistics. The shape of the exposure-response functions also strongly influenced predictions, reinforcing the need for contextually explicit experiments and improved mechanistic understanding of the processes driving behavioral and physiological responses to disturbance. This study presents a robust approach for integrating different types of empirical information to assess the effects of disturbance at spatio-temporal and ecological scales that are relevant to management and conservation.


Dual-task prioritization during overground and treadmill walking in healthy adults.
The dual-task effect on walking performance is different during treadmill and overground walking, though the cause of this difference is unknown. This study examined the effects of task prioritization on overground and treadmill dual-task walking. Twenty-two adults walked overground and on a treadmill under three dual-task conditions: prioritization of walking performance, prioritization of cognitive performance (serial subtraction in sevens), or no prioritization. Compared to single-task walking, stride velocity was reduced and stride time variability was increased during dual-task overground walking. During treadmill walking, there was no dual-task effect on walking performance, but cognitive task performance was improved. Prioritization of the cognitive task reduced the dual-task effect on stride velocity during overground walking only, whilst prioritization of the walking task reduced cognitive task performance in both walking modalities. These results corroborate recent findings that the dual-task effects on treadmill walking are not equivalent to those on overground walking. Healthy adults appear to prioritize cognitive task performance during treadmill dual-task walking without detrimental effects to gait. During overground walking however, allocation of attention to the secondary task reduces gait performance. These results indicate that treadmill based dual-task paradigms should not be used to infer factors which influence the cognitive control of overground walking.


Population polymorphism of nuclear mitochondrial DNA insertions reveals widespread diploidy associated with loss of heterozygosity in Debaryomyces hansenii.
Debaryomyces hansenii, a yeast that participates in the elaboration of foodstuff, displays important genetic diversity. Our recent phylogenetic classification of this species led to the subdivision of the species into three distinct clades. D. hansenii harbors the highest number of nuclear mitochondrial DNA (NUMT) insertions known so far for hemiascomycetous yeasts. Here we assessed the intraspecific variability of the NUMTs in this species by testing their presence/absence first in 28 strains, with 21 loci previously detected in the completely sequenced strain CBS 767(T), and second in a larger panel of 77 strains, with 8 most informative loci. We were able for the first time to structure populations in D. hansenii, although we observed little NUMT insertion variability within the clades. We determined the chronology of the NUMT insertions, which turned out to correlate with the previously defined taxonomy and provided additional evidence that colonization of nuclear genomes by mitochondrial DNA is a dynamic process in yeast. In combination with flow cytometry experiments, the NUMT analysis revealed the existence of both haploid and diploid strains, the latter being heterozygous and resulting from at least four crosses among strains from the various clades. As in the diploid pathogen Candida albicans, to which D. hansenii is phylogenetically related, we observed a differential loss of heterozygosity in the diploid strains, which can explain some of the large genetic diversity found in D. hansenii over the years.


Phosphorylation of 3'-azido-3'-deoxythymidine and selective interaction of the 5'-triphosphate with human immunodeficiency virus reverse transcriptase.
The thymidine analog 3'-azido-3'-deoxythymidine (BW A509U, azidothymidine) can inhibit human immunodeficiency virus (HIV) replication effectively in the 50-500 nM range [Mitsuya, H., Weinhold, K. J., Furman, P. A., St. Clair, M. H., Nusinoff-Lehrman, S., Gallo, R. C., Bolognesi, D., Barry, D. W. & Broder, S. (1985) Proc. Natl. Acad. Sci. USA 82, 7096-7100]. In contrast, inhibition of the growth of uninfected human fibroblasts and lymphocytes has been observed only at concentrations above 1 mM. The nature of this selectivity was investigated. Azidothymidine anabolism to the 5'-mono-, di-, and -triphosphate derivatives was similar in uninfected and HIV-infected cells. The level of azidothymidine monophosphate was high, whereas the levels of the di- and triphosphate were low (less than or equal to 5 microM and less than or equal to 2 microM, respectively). Cytosolic thymidine kinase (EC 188.8.131.52) was responsible for phosphorylation of azidothymidine to its monophosphate. Purified thymidine kinase catalyzed the phosphorylations of thymidine and azidothymidine with apparent Km values of 2.9 microM and 3.0 microM. The maximal rate of phosphorylation with azidothymidine was equal to 60% of the rate with thymidine. Phosphorylation of azidothymidine monophosphate to the diphosphate also appeared to be catalyzed by a host-cell enzyme, thymidylate kinase (EC 126.96.36.199). The apparent Km value for azidothymidine monophosphate was 2-fold greater than the value for dTMP (8.6 microM vs. 4.1 microM), but the maximal phosphorylation rate was only 0.3% of the dTMP rate. These kinetic constants were consistent with the anabolism results and indicated that azidothymidine monophosphate is an alternative-substrate inhibitor of thymidylate kinase. This conclusion was reflected in the observation that cells incubated with azidothymidine had reduced intracellular levels of dTTP. IC50 (concentration of inhibitor that inhibits enzyme activity 50%) values were determined for azidothymidine triphosphate with HIV reverse transcriptase and with immortalized human lymphocyte (H9 cell) DNA polymerase alpha. Azidothymidine triphosphate competed about 100-fold better for the HIV reverse transcriptase than for the cellular DNA polymerase alpha. The results reported here suggest that azidothymidine is nonselectively phosphorylated but that the triphosphate derivative efficiently and selectively binds to the HIV reverse transcriptase. Incorporation of azidothymidylate into a growing DNA strand should terminate DNA elongation and thus inhibit DNA synthesis.


Safety and Efficacy of High-Flow Nasal Cannula Therapy in Preterm Infants: A Meta-analysis.
High-flow therapy is the most recent, and popular, mode of respiratory support in neonates. However, the evidence supporting its efficacy and safety has not yet been established. We conducted a systematic review and meta-analysis of clinical trials comparing efficacy and safety of high-flow therapy compared with other modes of noninvasive ventilation (NIV) in preterm infants. Articles were indexed by using Medline, Embase, Scopus, OpenSIGLE, Health Management Information Consortium, and Cochrane Central Register of Controlled Trials. Randomized or quasi-randomized clinical trials involving preterm infants, comparing high-flow therapy with other modes of NIV, and reporting extractable data on relevant outcomes, were selected. Data on efficacy, safety, and other common neonatal outcomes were extracted on predesigned forms. In this analysis, we included 1112 preterm infants, participating in 9 clinical trials. High-flow therapy was similar in efficacy to other modes of NIV in preterm infants when used as primary support (odds ratio of failure of therapy, 1.02 [95% confidence interval: 0.55 to 1.88]), as well as after extubation (1.09 [0.58 to 2.02]). There were no significant differences in odds of death (0.48 [0.18 to 1.24]) between the groups. Preterm infants supported on high-flow had significantly lower odds of nasal trauma (0.13 [0.02 to 0.69]). High-flow therapy appears to be similar in efficacy and safety to other conventional modes of NIV in preterm infants. It is associated with significantly lower odds of nasal trauma. Caution needs to be exercised in extreme preterm infants because of the paucity of published data.


Complex Routes of Nosocomial Vancomycin-Resistant Enterococcus faecium Transmission Revealed by Genome Sequencing.
Vancomycin-resistant Enterococcus faecium (VREfm) is a leading cause of nosocomial infection. Here, we describe the utility of whole-genome sequencing in defining nosocomial VREfm transmission. A retrospective study at a single hospital in the United Kingdom identified 342 patients with E. faecium bloodstream infection over 7 years. Of these, 293 patients had a stored isolate and formed the basis for the study. The first stored isolate from each case was sequenced (200 VREfm [197 vanA, 2 vanB, and 1 isolate containing both vanA and vanB], 93 vancomycin-susceptible E. faecium) and epidemiological data were collected. Genomes were also available for E. faecium associated with bloodstream infections in 15 patients in neighboring hospitals, and 456 patients across the United Kingdom and Ireland. The majority of infections in the 293 patients were hospital-acquired (n = 249) or healthcare-associated (n = 42). Phylogenetic analysis showed that 291 of 293 isolates resided in a hospital-associated clade that contained numerous discrete clusters of closely related isolates, indicative of multiple introductions into the hospital followed by clonal expansion associated with transmission. Fine-scale analysis of 6 exemplar phylogenetic clusters containing isolates from 93 patients (32%) identified complex transmission routes that spanned numerous wards and years, extending beyond the detection of conventional infection control. These contained both vancomycin-resistant and -susceptible isolates. We also identified closely related isolates from patients at Cambridge University Hospitals NHS Foundation Trust and regional and national hospitals, suggesting interhospital transmission. These findings provide important insights for infection control practice and signpost areas for interventions. We conclude that sequencing represents a powerful tool for the enhanced surveillance and control of nosocomial E. faecium transmission and infection.


Harnessing the power of personality assessment: subjective assessment predicts behaviour in horses.
Objective assessment of animal personality is typically time consuming, requiring the repeated measure of behavioural responses. By contrast, subjective assessment of personality allows information to be collected quickly by experienced caregivers. However, subjective assessment must predict behaviour to be valid. Comparisons of subjective assessments and behaviour have been made but often with methodological weaknesses and thus, limited success. Here we test the validity of a subjective assessment against a battery of behaviour tests in 146 horses (Equus caballus). Our first aim was to determine if subjective personality assessment could predict behaviour during behaviour testing. We made specific a priori predictions for how subjectively measured personality should relate to behaviour testing. We found that Extroversion predicted time to complete a handling test and refusal behaviour during this test. It also predicted minimum distance to a novel object. Neuroticism predicted how reactive an individual was to a sudden visual stimulus but not how quickly it recovered from this. Agreeableness did not predict any behaviour during testing. There were several unpredicted correlations between subjective measures and behaviour tests which we explore further. Our second aim was to combine data from the subjective assessment and behaviour tests to gain a more comprehensive understanding of personality. We found that the combination of methods provides new insights into horse behaviour. Furthermore, our data are consistent with the idea of horses showing different coping styles, a novel finding for this species.


Combination Therapy With Exenatide Plus Pioglitazone Versus Basal/Bolus Insulin in Patients With Poorly Controlled Type 2 Diabetes on Sulfonylurea Plus Metformin: The Qatar Study.
The Qatar Study was designed to examine the efficacy of combination therapy with exenatide plus pioglitazone versus basal/bolus insulin in patients with long-standing poorly controlled type 2 diabetes mellitus (T2DM) on metformin plus a sulfonylurea. The study randomized 231 patients with poorly controlled (HbA1c >7.5%, 58 mmol/mol) T2DM on a sulfonylurea plus metformin to receive 1) pioglitazone plus weekly exenatide (combination therapy) or 2) basal plus prandial insulin (insulin therapy) to maintain HbA1c <7.0% (53 mmol/mol). After a mean follow-up of 12 months, combination therapy caused a robust decrease in HbA1c from 10.0 ± 0.6% (86 ± 5.2 mmol/mol) at baseline to 6.1 ± 0.1% (43 ± 0.7 mmol/mol) compared with 7.1 ± 0.1% (54 ± 0.8 mmol/mol) in subjects receiving insulin therapy. Combination therapy was effective in lowering the HbA1c independent of sex, ethnicity, BMI, or baseline HbA1c. Subjects in the insulin therapy group experienced significantly greater weight gain and a threefold higher rate of hypoglycemia than patients in the combination therapy group. Combination exenatide/pioglitazone therapy is a very effective and safe therapeutic option in patients with long-standing poorly controlled T2DM on metformin plus a sulfonylurea.


The recurrent domestication of viruses: major evolutionary transitions in parasitic wasps.
Several lineages of endoparasitoid wasps, which develop inside the body of other insects, have domesticated viruses, used as delivery tools of essential virulence factors for the successful development of their progeny. Virus domestications are major evolutionary transitions in highly diverse parasitoid wasps. Much progress has recently been made to characterize the nature of these ancestrally captured endogenous viruses that have evolved within the wasp genomes. Virus domestication from different viral families occurred at least three times in parasitoid wasps. This evolutionary convergence led to different strategies. Polydnaviruses (PDVs) are viral gene transfer agents and virus-like particles of the wasp Venturia canescens deliver proteins. Here, we take the standpoint of parasitoid wasps to review current knowledge on virus domestications by different parasitoid lineages. Then, based on genomic data from parasitoid wasps, PDVs and exogenous viruses, we discuss the different evolutionary steps required to transform viruses into vehicles for the delivery of the virulence molecules that we observe today. Finally, we discuss how endoparasitoid wasps manipulate host physiology and ensure parasitism success, to highlight the possible advantages of viral domestication as compared with other virulence strategies.


High-throughput Phenotyping of Lung Cancer Somatic Mutations.
Recent genome sequencing efforts have identified millions of somatic mutations in cancer. However, the functional impact of most variants is poorly understood. Here we characterize 194 somatic mutations identified in primary lung adenocarcinomas. We present an expression-based variant-impact phenotyping (eVIP) method that uses gene expression changes to distinguish impactful from neutral somatic mutations. eVIP identified 69% of mutations analyzed as impactful and 31% as functionally neutral. A subset of the impactful mutations induces xenograft tumor formation in mice and/or confers resistance to cellular EGFR inhibition. Among these impactful variants are rare somatic, clinically actionable variants including EGFR S645C, ARAF S214C and S214F, ERBB2 S418T, and multiple BRAF variants, demonstrating that rare mutations can be functionally important in cancer.


Exposure to bisphenol A is directly associated with inflammation in healthy Korean adults.
It was recently discovered that bisphenol A (BPA) and phthalates are cardiovascular disruptors. Inflammation is central to the initiation and progression of cardiovascular disease (CVD). This study evaluated whether BPA and different phthalate metabolites are associated with the inflammation marker high-sensitivity C-reactive protein (hs-CRP) in healthy Korean adults. This research is part of an ongoing, population-based study of Korean adults (30-64 years of age) conducted at the Cardiovascular and Metabolic Diseases Etiology Research Center (CMERC). The study enrolled 200 healthy volunteers (96 men, 104 women). Plasma hs-CRP was measured as an inflammation marker. BPA and five phthalate metabolites in urine were analyzed by using liquid chromatography/tandem mass spectrometry. BPA and monobenzyl phthalate (MBzP) differed significantly between the low-hs-CRP (<2 mg/L) and high-hs-CRP (≥2 mg/L) groups. BPA and MBzP were related to hs-CRP in an inverted L-shaped manner. High BPA levels (≥75th percentile) had significant odd ratios (ORs) for high hs-CRP even after adjusting for confounding factors related to obesity and insulin resistance, such as visceral fat volume, body mass index (BMI), adiponectin, high-density lipoprotein (HDL) cholesterol, hemoglobin A1c (HbA1c), and homeostasis model assessment of insulin resistance (HOMA-IR) (OR = 2.85; 95 % CI, 1.16-6.97). However, there was no significant association for MBzP ≥75th percentile. BPA was significantly related to high hs-CRP, even after adjusting for factors related to obesity and insulin resistance. Therefore, BPA could have a direct relationship with systemic inflammation regardless of obesity or insulin resistance.


Functional structures of the recA protein found by chimera analysis.
We developed a novel genetic method for finding functional regions of a protein by the analysis of chimeras formed between homologous proteins. Sets of chimeric genes were made by intramolecular homologous recombination in a linearized plasmid DNA carrying both recA genes of Escherichia coli and Pseudomonas aeruginosa. A recBCsbcA strain of E. coli was used for isolation of plasmids carrying recombinants between these genes. Examination of properties of E. coli strains deleting the recA gene and carrying a plasmid with a chimeric gene shows that chimera formation at certain positions inactivates a RecA function. Frequently, all chimeras with a junction in a certain region of the protein inactivate a function. Rather than a direct effect of the presence of the junction at a particular position, mismatching of the regions both sides of the junction that are derived from the different species is responsible for the inactivation. For a chimeric protein to be functional, certain pairs of sequences in different regions of the protein must derive from the same parent. Four pairs of such sequences were found: two are involved in activities for genetic recombination and for resistance to ultraviolet light irradiation and the others in formation of active oligomers. Regions defined by these sequences are located in the looped regions of the protein. A pair of regions may co-operate to form a functional folded structure.


Mass chemotherapy with diethylcarbamazine for the control of Bancroftian filariasis: a twelve-year follow-up in northern Trinidad, including observations on Mansonella ozzardi.
A microfilaria survey was conducted in Trinidad in 1992, 12 years after mass treatment with spaced doses of diethylcarbamazine citrate (DEC-C) for the control of Bancroftian filariasis; 348 persons were examined using thick blood smears and a membrane filtration technique. They included 104 who had participated in the mass chemotherapy campaign in 1980. No Wuchereria bancrofti microfilariae were detected among 66% of the population examined. In 1980, 86 of 592 persons examined were found to be infected with W. bancrofti, 140 with Mansonella ozzardi and 44 with mixed infections, while in 1992, only M. ozzardi infections persisted despite treatment with DEC-C. Of the 104 persons reexamined 12 years later, 46 had M. ozzardi, of which five were new cases, but none had W. bancrofti. During both the 1980 and 1992 surveys, low microfilariae rates for M. ozzardi were observed among those 19 years of age or younger. Of the 302 persons newly examined in 1992, 29 were infected with significantly (P < 0.001) more males (79.3%) than females (20.9%) being microfilaremic. The combined results showed similar prevalence rates for M. ozzardi from 23.3% to 21.6% in 1980 and 1992. Nuclepore membrane filtration and thick blood films were very efficient in demonstrating the presence of microfilariae. The usefulness of these methods and spaced treatment using DEC-C are also discussed.


A headspace needle-trap method for the analysis of volatile organic compounds in whole blood.
Needle trap devices (NTDs) are a relatively new and promising tool for headspace (HS) analysis. In this study, a dynamic HS sampling procedure is evaluated for the determination of volatile organic compounds (VOCs) in whole blood samples. A full factorial design was used to evaluate the influence of the number of cycles and incubation time and it is demonstrated that the controlling factor in the process is the number of cycles. A mathematical model can be used to determine the most appropriate number of cycles required to adsorb a prefixed amount of VOCs present in the HS phase whenever quantitative adsorption is reached in each cycle. Matrix effect is of great importance when complex biological samples, such as blood, are analyzed. The evaluation of the salting out effect showed a significant improvement in the volatilization of VOCs to the HS in this type of matrices. Moreover, a 1:4 (blood:water) dilution is required to obtain quantitative recoveries of the target analytes when external calibration is used. The method developed gives detection limits in the 0.020-0.080μg L(-1) range (0.1-0.4μg L(-1) range for undiluted blood samples) with appropriate repeatability values (RSD<15% at high level and <23% at LOQ level). Figure of merits of the method can be improved by using a smaller phase ratio (i.e., an increase in the blood volume and a decrease in the HS volume), which lead to lower detection limits, better repeatability values and greater sensibility. Twenty-eight blood samples have been evaluated with the proposed method and the results agree with those indicated in other studies. Benzene was the only target compound that gave significant differences between blood levels detected in volunteer non-smokers and smokers.


Influence of polymer architecture and molecular weight of poly(2-(dimethylamino)ethyl methacrylate) polycations on transfection efficiency and cell viability in gene delivery.
Nonviral gene delivery with the help of polycations has raised considerable interest in the scientific community over the past decades. Herein, we present a systematic study on the influence of the molecular weight and architecture of poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) on the transfection efficiency and the cytotoxicity in CHO-K1 cells. A library of well-defined homopolymers with a linear and star-shaped topology (3- and 5-arm stars) was synthesized via atom transfer radical polymerization (ATRP). The molecular weights of the polycations ranged from 16 to 158 kDa. We found that the cytotoxicity at a given molecular weight decreased with increasing number of arms. For a successful transfection a minimum molecular weight was necessary, since the polymers with a number-average molecular weight, M(n), below 20 kDa showed negligible transfection efficiency at any of the tested polyelectrolyte complex compositions. From the combined analysis of cytotoxicity and transfection data, we propose that polymers with a branched architecture and an intermediate molecular weight are the most promising candidates for efficient gene delivery, since they combine low cytotoxicity with acceptable transfection results.


Visual and health outcomes, measured with the activity inventory and the EQ-5D, in visual impairment.
Generic instruments to assess health utilities can be used to express the burden of health problems in widely used indexes. That is in contrast with what can be obtained with condition-specific instruments, outcomes are very specific and difficult to compare across conditions. The purpose of this study was to assess health and visual outcomes and its determinants in patients with visual impairment (VI) using the EQ-5D-3L and the Activity Inventory (AI). Participants were recruited in different hospitals during the PCVIP-study. A total of 134 patients with acuity 0.30 logMAR or less in the better eye were interviewed. The AI includes 46 goals split between three objectives: social functioning, recreation and daily living, and was used to measure visual ability. The EQ-5D consists of five questions covering one domain each and was used to provide a measure of health states. Responses to each domain were combined to produce a single individual index. The AI and the EQ-5D-3L showed enough discriminatory power between VI levels (p < 0.001), and their results were strongly correlated r(134) = 0.825, (p < 0.001). Explanatory factors for visual ability were level of VI in better eye, age and gender, R2 = 0.43, (p < 0.001). Explanatory factors for the EQ-5D-3L were level of VI in the better eye, comorbidities and gender, R2 = 0.36, (p < 0.001). Our results showed that the EQ-5D-3L is useful when characterizing the burden of VI and to compute, when necessary, quality-adjusted-life-years (QALY) changes due to VI. However, it is important to consider that the EQ-5D-3L uses a coarse response scale, assesses a limited spectrum of domains and is influenced by comorbidities. This might limit its responsiveness to small changes in visual ability.


Regioselectivity of the Pauson-Khand reaction in single-walled carbon nanotubes.
Chemical functionalization of nanotubes, in which their properties can be combined with those of other classes of materials, is fundamental to improve the physicochemical properties of nanotubes for potential technological applications. In this work, we theoretically and experimentally examine the Pauson-Khand reaction (PKR) on zig-zag, armchair, and chiral single-walled carbon nanotubes (SWCNTs). Our benchmarked density functional theory (DFT) calculations show that an alternative pathway to the widely accepted Magnus reaction pathway has significantly lower energy barriers, thus suggesting the use of this alternative pathway to predict whether a PKR on SWCNTs is favored or hampered. Accessible energy barriers of up to 16 kcal mol-1 are estimated and our results suggest that semiconducting SWCNTs react faster than metallic ones, although both types can be functionalized. Guided by our theoretical predictions, cyclopentenones are successfully attached to SWCNTs by heating and are, subsequently, characterized in the laboratory.


[Significance of cholesterol levels in patients 75 years or older].
Increasing comorbidity with aging reduces the predictive power of cardiovascular risk factors. From the age of 70 onward, total cholesterol levels decrease, perhaps associated with changes in the composition of some lipoprotein fractions. In subjects older than 75 years, being in the lowest quartile of cholesterol, insulinemia or serum albumin concentrations is associated with increased mortality. Cholesterol levels below 189 mg/dL in subjects older than 75 years should be considered an early sign of unidentified comorbidity or of rapid functional decline. HDL cholesterol levels, rather than total or LDL cholesterol, were inversely associated with increased mortality from ischemic coronary disease and stroke appears to rise as HDL cholesterol levels fall, rather than total or LDL cholesterol. On the other hand, LDL concentrations below 106 mg/dL and HDL concentrations below 36 mg/dL were associated with an increased risk of death from infectious disease. Stroke incidence, in particular, ischemic stroke, is highest in subjects older than 75 years. HDL cholesterol levels above 35 mg/dL appear to have a protective effect against ischemic stroke in subjects younger than 70 years. Two interventional drug studies investigating the effects of two statins (simvastatin and pravastatin) found that in subgroups of subjects older than 75 these drugs were associated with a reduction in all-cause mortality and cardiovascular morbidity, regardless of total cholesterol levels, but had no short-term effect on cognitive function.


Genetically timed, activity-sensor and rainbow transsynaptic viral tools.
We developed retrograde, transsynaptic pseudorabies viruses (PRVs) with genetically encoded activity sensors that optically report the activity of connected neurons among spatially intermingled neurons in the brain. Next we engineered PRVs to express two differentially colored fluorescent proteins in a time-shifted manner to define a time period early after infection to investigate neural activity. Finally we used multiple-colored PRVs to differentiate and dissect the complex architecture of brain regions.


Hsd11b2 haploinsufficiency in mice causes salt sensitivity of blood pressure.
Salt sensitivity of blood pressure is an independent risk factor for cardiovascular morbidity. Mechanistically, abnormal mineralocorticoid action and subclinical renal impairment may blunt the natriuretic response to high sodium intake, causing blood pressure to rise. 11β-Hydroxysteroid dehydrogenase type 2 (11βHSD2) controls ligand access to the mineralocorticoid receptor, and ablation of the enzyme causes severe hypertension. Polymorphisms in HSD11B2 are associated with salt sensitivity of blood pressure in normotensives. In this study, we used mice heterozygote for a null mutation in Hsd11b2 (Hsd11b2(+/-)) to define the mechanisms linking reduced enzyme activity to salt sensitivity of blood pressure. A high-sodium diet caused a rapid and sustained increase in blood pressure in Hsd11b2(+/-) mice but not in wild-type littermates. During the adaptation to high-sodium diet, heterozygotes displayed impaired sodium excretion, a transient positive sodium balance, and hypokalemia. After 21 days of high-sodium feeding, Hsd11b2(+/-) mice had an increased heart weight. Mineralocorticoid receptor antagonism partially prevented the increase in heart weight but not the increase in blood pressure. Glucocorticoid receptor antagonism prevented the rise in blood pressure. In Hsd11b2(+/-) mice, high-sodium feeding caused suppression of aldosterone and a moderate but sustained increase in corticosterone. This study demonstrates an inverse relationship among 11βHSD2 activity, heart weight, and blood pressure in a clinically important context. Reduced activity causes salt sensitivity of blood pressure, but this does not reflect illicit activation of mineralocorticoid receptors by glucocorticoids. Instead, we have identified a novel interaction among 11βHSD2, dietary salt, and circulating glucocorticoids.


Predictors of health decline in older adults with pneumonia: findings from the Community Acquired Pneumonia Impact Study.
The purpose of this study was to identify predictors of health decline among older adults with clinically diagnosed community acquired pneumonia (CAP). It was hypothesized that older adults with CAP who had lower levels of social support would be more likely to report a decline in health. A telephone survey was used to collect detailed information from older adults about their experiences with CAP. A broader determinants of health framework was used to guide data collection. This was a community wide study with participants being recruited from all radiology clinics in one Ontario community. The most important predictors of a health decline included: two symptoms (no energy; diaphoresis), two lifestyle variables (being very active; allowing people to smoke in their home), one quality of life variable (little difficulty in doing usual daily activities) and one social support variable (having siblings). A multiplicity of factors was found to be associated with a decline in health among older adults with clinically diagnosed CAP. These findings may be useful to physicians, family caregivers and others for screening older adults and providing interventions to help ensure positive health outcomes.