Effect of fish oil on atherogenesis in Watanabe heritable hyperlipidemic rabbit.
The effect of dietary supplementation with fish oil was investigated in the Watanabe heritable hyperlipidemic (WHHL) rabbit. Rabbits were fed Maxepa (1 g/kg body weight) from weaning for a 6-month period and were compared to controls. Fish oil supplements resulted in significant decreases in plasma triglyceride and cholesterol at 4 weeks after the start of the study, and these differences persisted over the 6-month period. No differences in the distribution of cholesterol or triglyceride among lipoprotein particles were found, nor were there differences in low density lipoprotein particle size. The decreases in plasma cholesterol and triglyceride were accompanied by significant increases in the percent of eicosapentaenoic and docosahexaenoic acids in both plasma and platelets. Fish oil supplementation resulted in a decrease in platelet count and response to the aggregating agents adenosine diphosphate, arachidonic acid, and collagen. Systolic blood pressure was significantly lower in the rabbits fed fish oil. At the end of the study period, aortic free and esterified cholesterol and intimal surface area covered with plaque were determined in the arch and in the descending and abdominal aorta. Significantly less cholesterol was found in the descending and abdominal aorta. These differences were mirrored by decreased activities of acid lipase and N-acetyl-beta-glucosaminidase. In contrast, no differences in the intimal surface area covered with plaque were found. In conclusion, dietary fish oil resulted in a decrease in aortic lipid deposition in the WHHL rabbit, which may have been due to decreases in plasma triglyceride and cholesterol, platelet count and aggregability, and systolic blood pressure.


On Singularities and Black Holes in Combination-Driven Models of Technological Innovation Networks.
It has been suggested that innovations occur mainly by combination: the more inventions accumulate, the higher the probability that new inventions are obtained from previous designs. Additionally, it has been conjectured that the combinatorial nature of innovations naturally leads to a singularity: at some finite time, the number of innovations should diverge. Although these ideas are certainly appealing, no general models have been yet developed to test the conditions under which combinatorial technology should become explosive. Here we present a generalised model of technological evolution that takes into account two major properties: the number of previous technologies needed to create a novel one and how rapidly technology ages. Two different models of combinatorial growth are considered, involving different forms of ageing. When long-range memory is used and thus old inventions are available for novel innovations, singularities can emerge under some conditions with two phases separated by a critical boundary. If the ageing has a characteristic time scale, it is shown that no singularities will be observed. Instead, a "black hole" of old innovations appears and expands in time, making the rate of invention creation slow down into a linear regime.


Lovastatin induces the formation of abnormal myelin-like membrane sheets in primary oligodendrocytes.
Statins, well-known inhibitors of cholesterol synthesis and protein isoprenylation, have been proposed as therapeutic drugs for multiple sclerosis (MS). As lovastatin and simvastatin, which are currently tested for their use in MS, can cross the blood-brain barrier, they may affect cellular processes in the central nervous system. This is especially relevant with respect to remyelination as a proposed additional treatment for MS, because cholesterol is a major component of myelin. Here, we show that primary oligodendrocytes, treated with lovastatin, form extensive membrane sheets, which contain galactosphingolipids. However, these membrane sheets are devoid of the major myelin proteins, myelin basic protein (MBP) and proteolipid protein (PLP). Reduced MBP protein expression was confirmed by SDS-PAGE and Western blotting, and in situ hybridization experiments revealed that lovastatin blocks MBP mRNA transport into oligodendrocyte processes. In contrast, PLP expression was only mildly affected by lovastatin. However, lovastatin treatment resulted in intracellular accumulation of PLP and prevented its translocation to the cell surface. Interestingly, another inhibitor of cholesterol synthesis (ro48-8071), which does not interfere with isoprenylation, had a similar effect on the localization of PLP, but it did not affect MBP expression and localization. These results suggest that lovastatin affects PLP transport predominantly by the inhibition of cholesterol synthesis, whereas reduced MBP expression is caused by impaired isoprenylation. Based on these results we recommend to carefully monitor the effect of statins on myelination prior to their use in demyelinating diseases.


Iterative phase retrieval strategy of transient events.
Important nuances of a process or processes in action can be obtained from the phase retrieval of diffraction patterns for analysis of transient events. A significant limitation associated with the iterative approach is that predictive input functions are needed and can result in situations of nonconvergence. In dealing with a transient event recorded as a series of Fourier magnitude patterns, such a hit-and-miss characteristic, on the surface, appears computationally daunting. We report and demonstrate a strategy here that effectively minimizes this by using a prior retrieved frame as the predictive function for the current retrieval process.


Effects of age on the detection and management of breast cancer.
Currently, breast cancer affects approximately 12% of women worldwide. While the incidence of breast cancer rises with age, a younger age at diagnosis is linked to increased mortality. We discuss age related factors affecting breast cancer diagnosis, management and treatment, exploring key concepts and identifying critical areas requiring further research. We examine age as a factor in breast cancer diagnosis and treatment relating it to factors such as genetic status, breast cancer subtype, hormone factors and nodal status. We examine the effects of age as seen through the adoption of population wide breast cancer screening programs. Assessing the incidence rates of each breast cancer subtype, in the context of age, we examine the observed correlations. We explore how age affects patient's prognosis, exploring the effects of age on stage and subtype incidence. Finally we discuss the future of breast cancer diagnosis and treatment, examining the potential of emerging tests and technologies (such as microRNA) and how novel research findings are being translated into clinically relevant practices.


Use of antiemetic agents in acute gastroenteritis: a systematic review and meta-analysis.
To perform a systematic review and meta-analysis to determine whether taking antiemetic drugs reduces vomiting and decreases the need for further intervention in children with gastroenteritis without causing significant adverse effects. Computerized databases, reference lists, and expert recommendations. Prospective controlled trials evaluating medication use in children with vomiting from gastroenteritis. Antiemetic drug therapy. Emesis cessation, use of intravenous fluid for rehydration, hospital admission, return to care, and medication adverse effects. The 11 articles that met the inclusion criteria evaluated various antiemetic agents: ondansetron (n = 6), domperidone (n = 2), trimethobenzamide (n = 2), pyrilamine-pentobarbital (n = 2), metoclopramide (n = 2), dexamethasone (n = 1), and promethazine (n = 1). Meta-analysis of 6 randomized, double-masked, placebo-controlled trials of ondansetron demonstrated decreased risk of further vomiting (5 studies; relative risk [RR], 0.45; 95% confidence interval [CI], 0.33-0.62; number needed to treat [NNT] = 5), reduced need for intravenous fluid (4 studies; RR, 0.41; 95% CI, 0.28-0.62; NNT = 5), and decreased risk of immediate hospital admission (5 studies; RR, 0.52; 95% CI, 0.27-0.95; NNT = 14). Diarrheal episodes increased in ondansetron-treated patients in 3 studies. Ondansetron use did not significantly affect return to care (5 studies; RR, 1.34; 95% CI, 0.77-2.35). Ondansetron therapy decreases the risk of persistent vomiting, the use of intravenous fluid, and hospital admissions in children with vomiting due to gastroenteritis. Future treatment guidelines should incorporate ondansetron therapy for select children with gastroenteritis.


Vitamin D and omega-3 polyunsaturated fatty acid supplementation in athletes with exercise-induced bronchoconstriction: a pilot study.
The aim of this pilot study was to determine the combined effect of vitamin D and omega-3 polyunsaturated fatty acid (PUFA) supplementation on airway function and inflammation in recreational athletes with exercise-induced bronchoconstriction (EIB). Ten recreational athletes with EIB participated in a single-blind, placebo-controlled trial over six consecutive weeks. All subjects attended the laboratory on three occasions. Each visit was separated by a period of 3 weeks: visit 1 (usual diet), visit 2 (placebo) and visit 3 (SMARTFISH® NutriFriend 2000; 30 µg vitamin D3-3000 mg eicosapentaenoic acid, 3000 mg docosahexaenoic acid) consumed once daily for a period of 3 weeks. Venous blood was collected at the beginning of each trial to determine vitamin D status. Spirometry was performed pre- and post-eucapnic voluntary hyperpnoea (EVH). The Maximum fall in FEV1 (ΔFEV1max) post-EVH was not different between visits (usual diet: -15.9 ± 3.6%, placebo: -16.1 ± 6.1%, vitamin D + omega-3 PUFA: -17.8 ± 7.2%). Serum vitamin D remained unchanged between visits. Vitamin D and omega-3 PUFA supplementation does not attenuate the reduction in lung function post-EVH. This finding should be viewed as preliminary until the results of randomised controlled trials are made available.


Role of protein tyrosine phosphatase 1B in vascular endothelial growth factor signaling and cell-cell adhesions in endothelial cells.
Vascular endothelial growth factor (VEGF) binding induces phosphorylation of VEGF receptor (VEGFR)2 in tyrosine, which is followed by disruption of VE-cadherin-mediated cell-cell contacts of endothelial cells (ECs), thereby stimulating EC proliferation and migration to promote angiogenesis. Tyrosine phosphorylation events are controlled by the balance of activation of protein tyrosine kinases and protein tyrosine phosphatases (PTPs). Little is known about the role of endogenous PTPs in VEGF signaling in ECs. In this study, we found that PTP1B expression and activity are markedly increased in mice hindlimb ischemia model of angiogenesis. In ECs, overexpression of PTP1B, but not catalytically inactive mutant PTP1B-C/S, inhibits VEGF-induced phosphorylation of VEGFR2 and extracellular signal-regulated kinase 1/2, as well as EC proliferation, whereas knockdown of PTP1B by small interfering RNA enhances these responses, suggesting that PTP1B negatively regulates VEGFR2 signaling in ECs. VEGF-induced p38 mitogen-activated protein kinase phosphorylation and EC migration are not affected by PTP1B overexpression or knockdown. In vivo dephosphorylation and cotransfection assays reveal that PTP1B binds to VEGFR2 cytoplasmic domain in vivo and directly dephosphorylates activated VEGFR2 immunoprecipitates from human umbilical vein endothelial cells. Overexpression of PTP1B stabilizes VE-cadherin-mediated cell-cell adhesions by reducing VE-cadherin tyrosine phosphorylation, whereas PTP1B small interfering RNA causes opposite effects with increasing endothelial permeability, as measured by transendothelial electric resistance. In summary, PTP1B negatively regulates VEGFR2 receptor activation via binding to the VEGFR2, as well as stabilizes cell-cell adhesions through reducing tyrosine phosphorylation of VE-cadherin. Induction of PTP1B by hindlimb ischemia may represent an important counterregulatory mechanism that blunts overactivation of VEGFR2 during angiogenesis in vivo.


Morphology and myofiber composition of skeletal musculature of the forelimb in young and aged wild type and myostatin null mice.
Most current research into therapeutic approaches to muscle diseases involves the use of the mouse as an experimental model. Furthermore, a major strategy to alleviate myopathic symptoms through enhancing muscle growth and regeneration is to inhibit the action of myostatin (Mstn), a transforming growth factor-beta (TGF-beta) family member that inhibits muscle growth. Presently, however, no study has expanded the morphological analysis of mouse skeletal muscle beyond a few individual muscles of the distal hindlimb, through which broad conclusions have been based. Therefore, we have initially undertaken an expansive analysis of the skeletal musculature of the mouse forelimb and highlighted the species-specific differences between equivalent muscles of the rat, another prominently used experimental model. Subsequently, we examined the musculature of the forelimb in both young and old adult wild-type (mstn(+/+)) and myostatin null (mstn(-/-)) mice and assessed the potential beneficial and detrimental effects of myostatin deletion on muscle morphology and composition during the aging process. We showed that: (1) the forelimb muscles of the mouse display a more glycolytic phenotype than those of the rat; (2) in the absence of myostatin, the induced myofiber hyperplasia, hypertrophy, and glycolytic conversion all occur in a muscle-specific manner; and, importantly, (3) the loss of myostatin significantly alters the dynamics of postnatal muscle growth and impairs age-related oxidative myofiber conversion.


Cardiovascular Events and Costs With Home Blood Pressure Telemonitoring and Pharmacist Management for Uncontrolled Hypertension.
Uncontrolled hypertension is a leading contributor to cardiovascular disease. A cluster-randomized trial in 16 primary care clinics showed that 12 months of home blood pressure telemonitoring and pharmacist management lowered blood pressure more than usual care (UC) for 24 months. We report cardiovascular events (nonfatal myocardial infarction, nonfatal stroke, hospitalized heart failure, coronary revascularization, and cardiovascular death) and costs over 5 years of follow-up. In the telemonitoring intervention (TI group, n=228), there were 15 cardiovascular events (5 myocardial infarction, 4 stroke, 5 heart failure, 1 cardiovascular death) among 10 patients. In UC group (n=222), there were 26 events (11 myocardial infarction, 12 stroke, 3 heart failure) among 19 patients. The cardiovascular composite end point incidence was 4.4% in the TI group versus 8.6% in the UC group (odds ratio, 0.49 [95% CI, 0.21-1.13], P=0.09). Including 2 coronary revascularizations in the TI group and 10 in the UC group, the secondary cardiovascular composite end point incidence was 5.3% in the TI group versus 10.4% in the UC group (odds ratio, 0.48 [95% CI, 0.22-1.08], P=0.08). Microsimulation modeling showed the difference in events far exceeded predictions based on observed blood pressure. Intervention costs (in 2017 US dollars) were $1511 per patient. Over 5 years, estimated event costs were $758 000 in the TI group and $1 538 000 in the UC group for a return on investment of 126% and a net cost savings of about $1900 per patient. Telemonitoring with pharmacist management lowered blood pressure and may have reduced costs by avoiding cardiovascular events over 5 years. Registration- URL: https://www.clinicaltrials.gov; Unique identifier: NCT00781365.


Hip abductor weakness in distance runners with iliotibial band syndrome.
To examine hip abductor strength in long-distance runners with iliotibial band syndrome (ITBS), comparing their injured-limb strength to their nonaffected limb and to the limbs of a control group of healthy long-distance runners; and to determine whether correction of strength deficits in the hip abductors of the affected runners through a rehabilitation program correlates with a successful return to running. Case series. Stanford University Sports Medicine Clinics. 24 distance runners with ITBS (14 female, 10 male) were randomly selected from patients presenting to our Runners' Injury Clinic with history and physical examination findings typical for ITBS. The control group of 30 distance runners (14 females, 16 males) were randomly selected from the Stanford University Cross-Country and Track teams. Group differences in hip abductor strength, as measured by torque generated, were analyzed using separate two-tailed t-tests between the injured limb, non-injured limb, and the noninjured limbs of the control group. Prerehabilitation hip abductor torque for the injured runners was then compared with postrehabilitation torque after a 6-week rehabilitation program. Hip abductor torque was measured with the Nicholas Manual Muscle Tester (kg), and normalized for differences in height and weight among subjects to units of percent body weight times height (%BWh). Average prerehabilitation hip abductor torque of the injured females was 7.82%BWh versus 9.82%BWh for their noninjured limb and 10.19%BWh for the control group of female runners. Average prerehabilitation hip abductor torque of the injured males was 6.86%BWh versus 8.62%BWh for their noninjured limb and 9.73%BWh for the control group of male runners. All prerehabilitation group differences were statistically significant at the p < 0.05 level. The injured runners were then enrolled in a 6-week standardized rehabilitation protocol with special attention directed to strengthening the gluteus medius. After rehabilitation, the females demonstrated an average increase in hip abductor torque of 34.9% in the injured limb, and the males an average increase of 51.4%. After 6 weeks of rehabilitation, 22 of 24 athletes were pain free with all exercises and able to return to running, and at 6-months follow-up there were no reports of recurrence. Long distance runners with ITBS have weaker hip abduction strength in the affected leg compared with their unaffected leg and unaffected long-distance runners. Additionally, symptom improvement with a successful return to the preinjury training program parallels improvement in hip abductor strength.


Synthesis engineering of iron oxide raspberry-shaped nanostructures.
Magnetic porous nanostructures consisting of oriented aggregates of iron oxide nanocrystals display very interesting properties such as a lower oxidation state of magnetite, and enhanced saturation magnetization in comparison with individual nanoparticles of similar sizes and porosity. However, the formation mechanism of these promising nanostructures is not well understood, which hampers the fine tuning of their magnetic properties, for instance by doping them with other elements. Therefore the formation mechanism of porous raspberry shaped nanostructures (RSNs) synthesized by a one-pot polyol solvothermal method has been investigated in detail from the early stages by using a wide panel of characterization techniques, and especially by performing original in situ HR-TEM studies in temperature. A time-resolved study showed the intermediate formation of an amorphous iron alkoxide phase with a plate-like lamellar structure (PLS). Then, the fine investigation of PLS transformation upon heating up to 500 °C confirmed that the synthesis of RSNs involves two iron precursors: the starting one (hydrated iron chlorides) and the in situ formed iron alkoxide precursor which decomposes with time and heating and contributes to the growth step of nanostructures. Such an understanding of the formation mechanism of RSNs is necessary to envision efficient and rational enhancement of their magnetic properties.


Contemporary pharmacological manipulation in assisted reproduction.
Follicle-stimulating hormone (FSH) treatment to induce follicular development in anovulating women and multiple follicular development for assisted conception has been incorporated in almost all reproductive treatment cycles in the form of either urinary, purified urinary or recombinant preparations. Besides improved tolerance and theoretically lower chances of infection by prions, the latter may be more effective in terms of clinical pregnancy rates, FSH requirement and cost effectiveness. The low-dose, step-up protocol to induce monofollicular development, which is applied worldwide, has to compete with the equally effective but health economically beneficial step-down protocol. The long protocol using recombinant FSH 150 IU/day is advocated when using gonadotropin-releasing hormone (GnRH) agonists in in vitro fertilisation (IVF) or intracytoplasmatic sperm injection treatment. However, the current paradigmatic hyperstimulation came under scrutiny after the introduction of the GnRH antagonists, which allow milder and more convenient approaches with acceptable cancellation and pregnancy rates but lower requirements for FSH. Risk of ovarian hyperstimulation syndrome (OHSS) can be further eliminated if recombinant luteinising hormone (rLH) or GnRH agonists are used to trigger oocyte maturation and ovulation; the latter require pituitary responsiveness and are therefore excluded in agonist protocols. FSH and LH are both required for appropriate folliculo- and steroidogenesis. In hypogonadotropic women, the addition of LH (human menopausal gonadotropin, human chorionic gonadotropin or rLH) is therefore obligate to achieve appropriate follicular growth and pregnancy. The role of LH in ovulation induction is still a matter of debate, although in GnRH agonistic protocols there seems to be a 'therapeutic window'; levels that are too high or too low have detrimental effects on IVF outcome. To broaden the pharmaceutical armoury, recent efforts have been directed towards the development of novel GnRH antagonists and FSH preparations with optimal pharmacokinetic, pharmacodynamic and safety profiles. Alternative strategies with fewer adverse effects and higher benefit/cost ratios are under development. However, before the GnRH agonist is abandoned for the antagonist as standard therapy, the cause of the observed possible lower pregnancy rates with the latter need to be clarified. In addition, prospective studies investigating possible direct effects of GnRH analogues, optimal dose-finding studies and treatment regimens under different conditions, with or without pharmacological coadministration and for different indications, should be performed to optimise the efficacy and tailor treatment strategies to individual needs.


Early prediction of favourable recovery 6 months after mild traumatic brain injury.
Predicting outcome after mild traumatic brain injury (MTBI) is notoriously difficult. Although it is recognised that milder head injuries do not necessarily mean better outcomes, less is known about the factors that do enable early identification of patients who are likely to recover well. To develop and internally validate two prediction rules for identifying patients who have the highest chance for good 6 month recovery. A prospective cohort study was conducted among patients with MTBI admitted to the emergency department. Apart from MTBI severity indices, a range of pre-, peri- and early post-injury variables were considered as potential predictors, including emotional and physical functioning. Logistic regression modelling was used to predict the absence of postconcussional symptoms (PCS) and full return to work (RTW). At follow-up, 64% of the 201 participating patients reported full recovery. Based on our prediction rules, patients without premorbid physical problems, low levels of PCS and post-traumatic stress early after injury had a 90% chance of remaining free of PCS. Patients with over 11 years of education, without nausea or vomiting on admission, with no additional extracranial injuries and only low levels of pain early after injury had a 90% chance of full RTW. The discriminative ability of the prediction models was satisfactory, with an area under the curve >0.70 after correction for optimism. Early identification of patients with MTBI who are likely to have good 6 month recovery was feasible on the basis of relatively simple prognostic models. A score chart was derived from the models to facilitate clinical application.


Gene transfer to the rabbit patellar tendon: potential for genetic enhancement of tendon and ligament healing.
Injuries to ligaments and tendons occur very commonly, but often heal very poorly. A number of growth factors show promise as biological agents with which to augment the reparative responses of these tissues, but they are extremely difficult to deliver clinically. We have suggested the use of gene transfer as a possible way to obviate this limitation. Here we have used the lacZ marker gene to evaluate methods for gene delivery to ligaments and tendons. Cell cultures were established from the following tissues of the New Zealand white rabbit: anterior and posterior cruciate ligaments, medial collateral ligament, patellar tendon and semitendionsus. In vitro experiments confirmed that these cells were very efficiently transduced by an E1- E3- adenoviral vector carrying the lacZ gene. Cells derived from ligaments and tendons also proved readily susceptible to retroviral transduction and, following transfer of the ned gene, selection in G418. To determine whether these cells could be genetically altered in situ, gene transfer to the patellar tendons of rabbits' knee joints was attempted using an adenovirus for in vivo delivery and a retrovirus for ex vivo delivery. Direct, percutaneous injection of the adenovirus resulted in high levels of lacZ expression, but this was limited to the area of tendon immediately adjacent to the injection site, and preferentially labeled cells in the epitenon sheath. An unexpected finding of these studies was the in vivo adenoviral transduction of bone cells in areas where the tendon insertion site had been accidentally injected. Allotransplantation of tendon fibroblasts into which the lacZ gene had been retrovirally introduced led to lacZ expression throughout the body of the tendon itself. The genetically altered cells appeared able to migrate from the site of injection and to integrate themselves into the crimp pattern of the tissue. Both in vivo and ex vivo methods led to gene expression for at least 6 weeks, which may be long enough for clinical purposes.


Changes in microbial contamination levels of porcine carcasses and fresh pork in slaughterhouses, processing lines, retail outlets, and local markets by commercial distribution.
The objective of this study was to evaluate the changes in microbial contamination levels of each porcine carcass and fresh pork in a general distribution process. A total of 100 commercial pigs were sampled (six sampling sites per individual, total 600 samples) at four sequential stages: slaughterhouse (after carcass grading and boning), processing line, retail outlet, and local market. No significant differences were observed in the contaminant percentages among sampling sites and sample collection years (P>0.05) with the exception of Bacillus cereus. The contaminant percentage of B. cereus at 1st collection year was higher than these of 2nd collection year (28.31% vs. 12.26%, P<0.05). B. cereus and Listeria monocytogenes were the most frequently detected pathogenic bacteria in the slaughterhouse and markets, respectively. On the other hand, Escherichia coli O157:H7 and Yersinia enterocolitica were not detected in carcasses or pork collected from any carcass sites and pork samples. However, the frequency of pathogenic bacteria in end-products at local markets was not highly related to the initial contamination of porcine carcasses in the slaughterhouse. Thus, the improvement of microbial safety for pork end-products requires hygienic control of porcine carcasses and meat cutting during all operations in the slaughterhouse, processing line, retail outlet, and local market.


[Two cases of giant metastatic cervical lymphnode from oral cancer responding to concurrent chemoradiotherapy with S-1].
We report two cases of giant metastatic cervical lymphnode from oral squamous cell carcinoma, successfully treated with concurrent chemoradiotherapy with S-1. Case 1 was a 80-year-old male who had an ipsilateral occult neck metastasis(52x46 mm mass)at level IV after surgery of the left tongue carcinoma(T2N0M0). Radiotherapy(2.0 Gy/day; 5 days/week)was given at a total dose of 66 Gy. Two courses of oral administration of S-1(61 mg/m2/day)for 2 weeks followed by 1-week rest period as one course was repeated with the concurrent radiotherapy. After the radiotherapy, the oral administration of S-1 alone was continued for 1 year under the same regimen. Case 2 was a 51-year-old male who had left tongue carcinoma(T4bN2cM0)with ipsilateral cervical lymphnode metastasis(88x44 mm mass). The same chemoradiotherapy(1.8 Gy/day; 5 days/week; a total dose of 63 Gy; S-1 53 mg/m2/day)was carried out as in case 1. These giant metastatic lymphnodes of case 1 and 2 disappeared by 3 and 6 months after radiotherapy, respectively, and achieved a complete response. This therapy may be effective not only for primary lesion of oral cancer, but also metastatic regional lymphnodes in some cases.


Describing self-care in Italian adults with heart failure and identifying determinants of poor self-care.
Self-care improves outcomes in patients with heart failure; however, no studies have been conducted on this topic in Italy. We aimed to describe self-care in Italian adults with heart failure and to identify sociodemographic and clinical determinants of self-care. A cross-sectional design was used to study 1192 heart failure patients enrolled across Italy. We measured self-care using the Self-Care of Heart Failure Index version 6.2, which measures self-care maintenance, management and confidence. Sociodemographic and clinical data were tested as potential determinants of self-care. The mean age of the sample was 72 (SD = 11) years; 58% were male. In the three areas of self-care, scores ranged from 53.18 to 55.26 and few people were adequate in self-care (14.5% to 24.4% of the sample). Self-care behaviours particularly low in this population were symptom monitoring, exercise, use of reminders to take medicines and symptom recognition. Confidence in the ability to keep oneself free of symptoms and relieve symptoms was low. Taking fewer medications, poor cognition, older age, having a caregiver, being male and having heart failure for a shorter time predicted poor self-care maintenance. Poor cognition, not being employed, being male, and having worse New York Heart Association class predicted poor self-care management. Poor cognition, taking fewer mediations, older age, and male gender predicted poor self-care confidence. Self-care is poor in Italian heart failure patients. Determinants of poor self-care identified in this study can help to target patients' education. Male gender and poor cognition were consistently associated with poor self-care maintenance, management and confidence.


Cardiovascular Health and Outcomes in Cancer Patients Receiving Immune Checkpoint Inhibitors.
Whether cardiovascular (CV) disease is associated with clinical outcomes in cancer patients receiving immunotherapy is unknown. We reviewed the Mayo Clinic database for all cancer patients who received an immune checkpoint inhibitor (ICI). Multivariate logistic regression analysis, survival analyses, and Cox proportional-hazards models were formulated. Between March, 2010 and July, 2019, 3,326 patients received ICI. Mean patient age was 63.5 years (range: 16 to 96 years). In a Cox proportional-hazards model, obesity (hazard ratio [HR] 0.65, 95% confidence level [CI] 0.55 to 0.77, p < 0.001) and hypercholesterolemia (HR 0.80, 95% CI 0.72 to 0.89, p < 0.001) were associated with lower all-cause mortality while hypertension (HR 1.32, 95% CI 1.17 to 1.49, p < 0.001) and smoking (HR 1.17, 95% CI 1.06 to 1.29, p = 0.002) were associated with higher overall mortality. Among patients with lung cancer, multivariable-adjusted hazard ratios for death from any cause for beta blocker users, as compared with patients who had never used a beta blocker, were 1.39 (95% CI 1.10 to 1.76, p = 0.006). A total of 80 patients (2.4%) experienced CV immune-related adverse events. Event-related morality for ICI-induced myocarditis was 41.7% (5/12). Multivariable-adjusted hazard ratios for ICI-induced myocarditis were 5.2 (95% CI 1.4 to 18.7, p = 0.01) for history of heart failure, 4.06 (95% CI 1.15 to 14.3, p = 0.03) for history of acute coronary syndrome, and 1.07 (per each 1-year increase, 95% CI 1.01 to 1.14, p = 0.02) for age. In conclusion, our study shows that CV factors are associated with clinical outcomes in cancer patients receiving ICI and could be used to predict mortality. In patients with lung cancer, pretreatment beta blocker use is associated with higher all-cause mortality. Three clinical factors-history of heart failure, history of acute coronary syndrome, and age greater than 80 years-help identify patients at higher risk of ICI-induced myocarditis who might benefit from more intensive cardiac surveillance.


Attenuation of opioid receptor activity by phorbol esters in neuroblastoma x glioma NG108-15 hybrid cells.
Chronic opioid treatment of neuroblastoma x glioma NG108-15 cells induces desensitization of the opioid receptor and this may involve a change in membrane protein phosphorylation. In an attempt to mimic this possible mechanism, we studied effects of phorbol ester activation of protein kinase C on opioid receptor activity. Incubation of NG108-15 hybrid cells with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) abolished up to 45% of opioid inhibition of cyclic AMP accumulation in intact cells, while basal accumulation and prostaglandin E1-stimulated cyclic AMP accumulation were unaltered. This decrease of opioid inhibition was dose- and time-dependent and the potency order of phorbol esters and apparent K activation (90 nM) for TPA were consistent with phorbol esters acting through the stimulation of protein kinase C. TPA also decreased the inhibition of cyclic AMP accumulation mediated through muscarinic and alpha-2 adrenergic receptors. These effects of TPA were best explained by a TPA-induced alteration of the inhibitory nucleotide-binding protein (Gi), the common transducer protein of these receptors. Impairment of Gi by TPA treatment was evidenced by a reduction in agonist-stimulated GTP hydrolysis and activation by GTP. Quantification of Gi by pertussis toxin-catalyzed ADP-ribosylation revealed that TPA decreased maximal labeling. In summary, phorbol esters appeared to attenuate opioid receptor activity by altering the activity of the transducer protein Gi.


RG7386, a Novel Tetravalent FAP-DR5 Antibody, Effectively Triggers FAP-Dependent, Avidity-Driven DR5 Hyperclustering and Tumor Cell Apoptosis.
Dysregulated cellular apoptosis and resistance to cell death are hallmarks of neoplastic initiation and disease progression. Therefore, the development of agents that overcome apoptosis dysregulation in tumor cells is an attractive therapeutic approach. Activation of the extrinsic apoptotic pathway is strongly dependent on death receptor (DR) hyperclustering on the cell surface. However, strategies to activate DR5 or DR4 through agonistic antibodies have had only limited clinical success. To pursue an alternative approach for tumor-targeted induction of apoptosis, we engineered a bispecific antibody (BsAb), which simultaneously targets fibroblast-activation protein (FAP) on cancer-associated fibroblasts in tumor stroma and DR5 on tumor cells. We hypothesized that bivalent binding to both FAP and DR5 leads to avidity-driven hyperclustering of DR5 and subsequently strong induction of apoptosis in tumor cells but not in normal cells. Here, we show that RG7386, an optimized FAP-DR5 BsAb, triggers potent tumor cell apoptosis in vitro and in vivo in preclinical tumor models with FAP-positive stroma. RG7386 antitumor efficacy was strictly FAP dependent, was independent of FcR cross-linking, and was superior to conventional DR5 antibodies. In combination with irinotecan or doxorubicin, FAP-DR5 treatment resulted in substantial tumor regression in patient-derived xenograft models. FAP-DR5 also demonstrated single-agent activity against FAP-expressing malignant cells, due to cross-binding of FAP and DR5 across tumor cells. Taken together, these data demonstrate that RG7386, a novel and potent antitumor agent in both mono- and combination therapies, overcomes limitations of previous DR5 antibodies and represents a promising approach to conquer tumor-associated resistance to apoptosis. Mol Cancer Ther; 15(5); 946-57. ©2016 AACR.


Post-traumatic administration of the p53 inactivator pifithrin-α oxygen analogue reduces hippocampal neuronal loss and improves cognitive deficits after experimental traumatic brain injury.
Traumatic brain injury (TBI) is a major cause of death and disability worldwide. Neuronal apoptosis in the hippocampus has been detected after TBI. The hippocampal dysfunction may result in cognitive deficits in learning, memory, and spatial information processing. Our previous studies demonstrated that a p53 inhibitor, pifithrin-α oxygen analogue (PFT-α (O)), significantly reduced cortical cell death, which is substantial following controlled cortical impact (CCI) TBI, and improved neurological functional outcomes via anti-apoptotic mechanisms. In the present study, we examined the effect of PFT-α (O) on CCI TBI-induced hippocampal cellular pathophysiology in light of this brain region's role in memory. To investigate whether p53-dependent apoptosis plays a role in hippocampal neuronal loss and associated cognitive deficits and to define underlying mechanisms, SD rats were subjected to experimental CCI TBI followed by the administration of PFT-α or PFT-α (O) (2mg/kg, i.v.) or vehicle at 5h after TBI. Magnetic resonance imaging (MRI) scans were acquired at 24h and 7days post-injury to assess evolving structural hippocampal damage. Fluoro-Jade C was used to stain hippocampal sub-regions, including CA1 and dentate gyrus (DG), for cellular degeneration. Neurological functions, including motor and recognition memory, were assessed by behavioral tests at 7days post injury. p53, p53 upregulated modulator of apoptosis (PUMA), 4-hydroxynonenal (4-HNE), cyclooxygenase-IV (COX IV), annexin V and NeuN were visualized by double immunofluorescence staining with cell-specific markers. Levels of mRNA encoding for caspase-3, p53, PUMA, Bcl-2, Bcl-2-associated X protein (BAX) and superoxide dismutase (SOD) were measured by RT-qPCR. Our results showed that post-injury administration of PFT-α and, particularly, PFT-α (O) at 5h dramatically reduced injury volumes in the ipsilateral hippocampus, improved motor outcomes, and ameliorated cognitive deficits at 7days after TBI, as evaluated by novel object recognition and open-field test. PFT-α and especially PFT-α (O) significantly reduced the number of FJC-positive cells in hippocampus CA1 and DG subregions, versus vehicle treatment, and significantly decreased caspase-3 and PUMA mRNA expression. PFT-α (O), but not PFT-α, treatment significantly lowered p53 and elevated SOD2 mRNA expression. Double immunofluorescence staining demonstrated that PFT-α (O) treatment decreased p53, annexin V and 4-HNE positive neurons in the hippocampal CA1 region. Furthermore, PUMA co-localization with the mitochondrial maker COX IV, and the upregulation of PUMA were inhibited by PFT-α (O) after TBI. Our data suggest that PFT-α and especially PFT-α (O) significantly reduce hippocampal neuronal degeneration, and ameliorate neurological and cognitive deficits in vivo via antiapoptotic and antioxidative properties.


Sex Differences in Autism Spectrum Disorder: An Investigation on Core Symptoms and Psychiatric Comorbidity in Preschoolers.
Findings regarding sex differences in autism spectrum disorder (ASD), as far as core symptoms and psychiatric comorbidities (PC) are concerned, are inconsistent, inconclusive, or conflicting among studies. The lower prevalence of ASD in females than in males and the age and intelligence quotient (IQ) heterogeneity among samples made it difficult to investigate these differences. This case-control study tries to deepen the impact of sex differences on core symptoms of autism and PC in 214 preschoolers with ASD (mean age, 45.26) without impairment in non-verbal IQ (nvIQ ≥70). A total of 107 ASD females (mean age, 44.51 ± 13.79 months) were matched one by one with 107 males (mean age, 46.01 ± 13.42 months) for chronological age (±6 months) and nvIQ (±6 points). We used the Autism Diagnostic Observation Schedule 2 (ADOS-2) and the Child Behavior Checklist (CBCL) 1.5-5 to explore autism severity and PC. The results highlight that ASD females did not significantly differ from ASD males regarding the severity of autism. Statistically significant lower levels of emotionally reactive (p = 0.005, η2 = 0.04), anxious-depressed (p = 0.001, η 2 = 0.05), internalizing problems (p = 0.04, η 2 = 0.02), and DSM-Oriented Scales anxiety problems (p = 0.02, η 2 = 0.04) in ASD females than in ASD males were also detected. Our findings of no difference in the autism severity and lower internalizing problems in females than males with ASD extend the knowledge of autism in females during preschool years. Compared to other similar studies on this topic, we can state that these results are not supported by differences in nvIQ between sexes nor by the presence of cognitive impairment. It confirms the need for clinicians to consider sex differences when describing autism psychopathology.


A conserved glutamate residue exhibits multifunctional catalytic roles in D-fructose-1,6-bisphosphate aldolases.
The aldolase catalytic cycle consists of a number of proton transfers that interconvert covalent enzyme intermediates. Glu-187 is a conserved amino acid that is located in the mammalian fructose-1,6-bisphosphate aldolase active site. Its central location, within hydrogen bonding distance of three other conserved active site residues: Lys-146, Glu-189, and Schiff base-forming Lys-229, makes it an ideal candidate for mediating proton transfers. Point mutations, Glu-187--> Gln, Ala, which would inhibit proton transfers significantly, compromise activity. Trapping of enzymatic intermediates in Glu-187 mutants defines a proton transfer role for Glu-187 in substrate cleavage and Schiff base formation. Structural data show that loss of Glu-187 negative charge results in hydrogen bond formation between Lys-146 and Lys-229 consistent with a basic pK(a) for Lys-229 in native enzyme and supporting nucleophilic activation of Lys-229 by Glu-187 during Schiff base formation. The crystal structures also substantiate Glu-187 and Glu-189 as present in ionized form in native enzyme, compatible with their role of catalyzing proton exchange with solvent as indicated from solvent isotope effects. The proton exchange mechanism ensures Glu-187 basicity throughout the catalytic cycle requisite for mediating proton transfer and electrostatic stabilization of ketamine intermediates. Glutamate general base catalysis is a recurrent evolutionary feature of Schiff base0forming aldolases.


Free-Form Diffractive Metagrating Design Based on Generative Adversarial Networks.
A key challenge in metasurface design is the development of algorithms that can effectively and efficiently produce high-performance devices. Design methods based on iterative optimization can push the performance limits of metasurfaces, but they require extensive computational resources that limit their implementation to small numbers of microscale devices. We show that generative neural networks can train from images of periodic, topology-optimized metagratings to produce high-efficiency, topologically complex devices operating over a broad range of deflection angles and wavelengths. Further iterative optimization of these designs yields devices with enhanced robustness and efficiencies, and these devices can be utilized as additional training data for network refinement. In this manner, generative networks can be trained, with a one-time computation cost, and used as a design tool to facilitate the production of near-optimal, topologically complex device designs. We envision that such data-driven design methodologies can apply to other physical sciences domains that require the design of functional elements operating across a wide parameter space.


Vertebral artery stenting following percutaneous transluminal angioplasty. Technical note.
The authors report initial results and follow up using stent placement to treat atherosclerotic stenosis in vertebral arteries. Three patients with severe atherosclerotic vascular disease underwent vertebral artery stent placement using a balloon expandable stent. Medical therapy (aspirin and warfarin) and conventional percutaneous angioplasty failed to resolve the disease and the patients developed symptomatic restenosis within 3 months of angioplasty. Two patients had symptoms of anterior circulation ischemia with carotid artery occlusions and reduced supply to the anterior circulation from the stenosed vertebral arteries. One patient had recurrent posterior circulation symptoms. Stents were successfully placed in all three, resulting in immediate reversal of stenosis and resolution of symptoms. Clinical follow-up study (mean 9 months) has shown no recurrent symptoms in the patient with posterior circulation symptoms, but the two patients with anterior circulation ischemia did develop recurrent symptoms. Angiographic follow up in these two patients at 3 months and 1 year, however, demonstrated continued patency of vertebral artery lumina. They underwent extracranial-intracranial bypass surgery to relieve their symptoms. This experience suggests stents can be placed without complication in the proximal vertebral arteries and may have an adjunctive role in the treatment of atherosclerotic cerebrovascular disease following unsuccessful angioplasty.


Cholinergic, GABAergic, and glutamate-enriched inputs from the mesopontine tegmentum to the subthalamic nucleus in the rat.
In order to clarify the origin and to examine the neurochemistry and synaptology of the projection from the mesopontine tegmentum (MTg) to the subthalamic nucleus (STN), rats received discrete deposits of anterograde tracers in different regions of the MTg. Anterogradely labeled fibers were examined in the light and electron microscopes. The distribution of GABA or glutamate immunoreactivity was examined by post-embedding immunocytochemistry. The anterograde tracing demonstrated that the projection to the STN arises from at least three divisions of the MTg: the area defined by the cholinergic neurons of the pedunculopontine region (PPN-Ch 5), the more medial and largely noncholinergic midbrain extrapyramidal area (MEA) and to a lesser extent the laterodorsal tegmental nucleus (LDTg). Post-embedding immunocytochemistry revealed that there are GABA-immunopositive and immunonegative components to this projection and at least a proportion of the GABA-immunonegative component is enriched in glutamate immunoreactivity. The similarity of the morphology, trajectory and synaptology of the anterogradely labeled fibers and the choline acetyltransferase (ChAT)-immunopositive fibers supports the proposal that at least part of the projection is cholinergic. The terminals anterogradely labeled from the MTg and the ChAT-immunoreactive terminals form asymmetrical synapses with the dendrites and spines of subthalamic neurons. Both anterogradely labeled and ChAT-positive terminals make convergent synaptic contacts with GABA-immunoreactive terminals that form symmetrical synaptic contacts and are probably derived from the globus pallidus. Taken together these findings imply that the MTg sends cholinergic, GABAergic and glutamatergic projections to the STN where at least one of the functional roles is to modulate the indirect pathway of information flow through the basal ganglia that is carried via the pallidosubthalamic projection.


Acute lung infections in normal and immunocompromised hosts.
Pulmonary infections are among the most common causes of morbidity and mortality worldwide, and contribute substantially to annual medical expenditures in the United States. Despite the availability of antimicrobial agents, pneumonia constitutes the sixth most common cause of death and the number one cause of death from infection. Pneumonia can be particularly life-threatening in the elderly, in individuals who have pre-existing heart and lung conditions, in patients who have suppressed or weakened immunity, and in pregnant women. This article discusses some of the important causes of acute lung infections in normal and immunocompromised hosts. Because there often is considerable overlap, infections are categorized by the host immune status that is most likely to be associated with a particular pathogen.


Lack of potential carcinogenicity for acesulfame potassium - Systematic evaluation and integration of mechanistic data into the totality of the evidence.
The safety of low- and no-calorie sweeteners remains a topic of general interest. Substantial evidence exists demonstrating a lack of carcinogenicity of the no-calorie sweetener acesulfame potassium (Ace K). The objective of this evaluation was to conduct a systematic assessment of available mechanistic data using a framework that quantitatively integrates proposed key characteristics of carcinogens (KCCs) into the totality of the evidence. Over 800 KCC-relevant endpoints from a variety of in vitro and in vivo assays were assessed for quality, relevance, and activity, and integrated to determine the overall strength of the evidence for plausibility that Ace K acts through the KCC. Overall, there was a lack of activity across the KCCs (overall integrated score <0 and no "strong" categorization for evidence of activity) in which data were identified. Together with the absence of treatment-related tumor effects in rodent bioassays, these results support the conclusion that Ace K is unlikely to induce a carcinogenic response. This assessment employed a weight of the evidence analysis that includes the consideration of factors such as reliability, strength of the model system, activity, and dose in a complex and heterogeneous dataset, and the ultimate integration of multiple data streams in the cancer hazard evaluation.


Cost and impact of scaling up interventions to save lives of mothers and children: taking South Africa closer to MDGs 4 and 5.
South Africa has made substantial progress on child and maternal mortality, yet many avoidable deaths of mothers and children still occur. This analysis identifies priority interventions to be scaled up nationally and projects the potential maternal and child lives saved. We modelled the impact of maternal, newborn and child interventions using the Lives Saved Tools Projections to 2015 and used realistic coverage increases based on expert opinion considering recent policy change, financial and resource inputs, and observed coverage change. A scenario analysis was undertaken to test the impact of increasing intervention coverage to 95%. By 2015, with realistic coverage, the maternal mortality ratio (MMR) can reduce to 153 deaths per 100,000 and child mortality to 34 deaths per 1,000 live births. Fifteen interventions, including labour and delivery management, early HIV treatment in pregnancy, prevention of mother-to-child transmission and handwashing with soap, will save an additional 9,000 newborns and children and 1,000 mothers annually. An additional US$370 million (US$7 per capita) will be required annually to scale up these interventions. When intervention coverage is increased to 95%, breastfeeding promotion becomes the top intervention, the MMR reduces to 116 and the child mortality ratio to 23. The 15 interventions identified were adopted by the National Department of Health, and the Health Minister launched a campaign to encourage Provincial Health Departments to scale up coverage. It is hoped that by focusing on implementing these 15 interventions at high quality, South Africa will reach Millennium Development Goal (MDG) 4 soon after 2015 and MDG 5 several years later. Focus on HIV and TB during early antenatal care is essential. Strategic gains could be realised by targeting vulnerable populations and districts with the worst health outcomes. The analysis demonstrates the usefulness of priority setting tools and the potential for evidence-based decision making in the health sector.


The pathogenesis of squamous cell cancer: lessons learned from studies of skin carcinogenesis--thirty-third G. H. A. Clowes Memorial Award Lecture.
The multistage nature of cancer pathogenesis was first defined over 50 years ago by the sequential topical application of chemical agents to mouse skin. Since then, the skin model has provided remarkable insights into the biology, biochemistry, pharmacology, and genetics of carcinogenesis. Discoveries from studies of mouse skin have proved to be landmarks in cancer research including: the binding of carcinogens to DNA; the monoclonal origin of benign and malignant tumors; the powerful tumor-promoting action of phorbol esters; the antipromoting potency of retinoids and steroids; the modifying role of age, caloric intake, and specific dietary constituents on cancer induction; the variable risk for benign tumors to progress to cancer; and the requirement for multiple genetic changes in malignant conversion. Many of these concepts are now widely applied to the interpretation of specific molecular discoveries both in simple experimental systems and in human cancers, but the power of this quantitative, multistage skin carcinogenesis model has made these assessments possible. It has also provided the separation of mechanistically distinct stages in cancer pathogenesis: initiation; promotion; premalignant progression; and malignant conversion. This paper will review our current understanding of the genetic, biological, and biochemical alterations that contribute to the evolution of each of these stages in skin carcinogenesis. These new insights provide an opportunity to replace the traditional operational-based schemata defining the process of carcinogenesis with a working model designed around functional alterations in neoplastic cells.


Organizational capacity for change in health care: Development and validation of a scale.
We do not have a strong understanding of a health care organization's capacity for attempting and completing multiple and sometimes competing change initiatives. Capacity for change implementation is a critical success factor as the health care industry is faced with ongoing demands for change and transformation because of technological advances, market forces, and regulatory environment. The aim of this study was to develop and validate a tool to measure health care organizations' capacity to change by building upon previous conceptualizations of absorptive capacity and organizational readiness for change. A multistep process was used to develop the organizational capacity for change survey. The survey was sent to two populations requesting answers to questions about the organization's leadership, culture, and technologies in use throughout the organization. Exploratory and confirmatory factor analyses were conducted to validate the survey as a measurement tool for organizational capacity for change in the health care setting. The resulting organizational capacity for change measurement tool proves to be a valid and reliable method of evaluating a hospital's capacity for change through the measurement of the population's perceptions related to leadership, culture, and organizational technologies. The organizational capacity for change measurement tool can help health care managers and leaders evaluate the capacity of employees, departments, and teams for change before large-scale implementation.


Investigation of interactions between TLR2, MyD88 and TIRAP by bioluminescence resonance energy transfer is hampered by artefacts of protein overexpression.
Toll like receptors (TLRs) are important pattern recognition receptors that can detect pathogen and danger associated molecular patterns to initiate an innate immune response. TLR1 and 2 heterodimerize at the plasma membrane upon binding to triacylated lipopeptides from bacterial cell walls, or to the synthetic ligand Pam3CSK4. TLR1/2 dimers interact with adaptor molecules TIRAP and MyD88 to initiate a signalling cascade that leads to activation of key transcription factors, including NF-kB. Despite TLRs being extensively studied over the last two decades, the real-time kinetics of ligand binding and receptor activation remains largely unexplored. We aimed to study the kinetics of TLR activation and recruitment of adaptors, using TLR1/2 dimer interactions with adaptors MyD88 and TIRAP. Bioluminescence resonance energy transfer (BRET) allows detection of real-time protein-protein interactions in living cells, and was applied to study adaptor recruitment to TLRs. Energy transfer showed interactions between TLR2 and TIRAP, and between TLR2 and MyD88 only in the presence of TIRAP. Quantitative BRET and confocal microscopy confirmed that TIRAP is necessary for MyD88 interaction with TLR2. Furthermore, constitutive proximity between the proteins in the absence of Pam3CSK4 stimulation was observed with BRET, and was not abrogated with lowered protein expression, changes in protein tagging strategies, or use of the brighter NanoLuc luciferase. However, co-immunoprecipitation studies did not demonstrate constitutive interaction between these proteins, suggesting that the interaction observed with BRET likely represents artefacts of protein overexpression. Thus, caution should be taken when utilizing protein overexpression in BRET studies and in investigations of the TLR pathway.


Limited Antigenic Diversity in Contemporary H7 Avian-Origin Influenza A Viruses from North America.
Subtype H7 avian-origin influenza A viruses (AIVs) have caused at least 500 confirmed human infections since 2003 and culling of >75 million birds in recent years. Here we antigenically and genetically characterized 93 AIV isolates from North America (85 from migratory waterfowl [1976-2010], 7 from domestic poultry [1971-2012], and 1 from a seal [1980]). The hemagglutinin gene of these H7 viruses are separated from those from Eurasia. Gradual accumulation of nucleotide and amino acid substitutions was observed in the hemagglutinin of H7 AIVs from waterfowl and domestic poultry. Genotype characterization suggested that H7 AIVs in wild birds form diverse and transient internal gene constellations. Serologic analyses showed that the 93 isolates cross-reacted with each other to different extents. Antigenic cartography showed that the average antigenic distance among them was 1.14 units (standard deviation [SD], 0.57 unit) and that antigenic diversity among the H7 isolates we tested was limited. Our results suggest that the continuous genetic evolution has not led to significant antigenic diversity for H7 AIVs from North America. These findings add to our understanding of the natural history of IAVs and will inform public health decision-making regarding the threat these viruses pose to humans and poultry.


Strategic planning for research use in nursing practice.
To prepare for a culture change to integrate research utilization into daily nursing practice, the authors conducted a descriptive survey of all registered nurses (RNs) in an integrated healthcare delivery system. The purposes of this study were to assess RNs' knowledge, attitudes, and practices (KAP) of nursing research activities, assess factors that support a research environment, and determine facilitating and challenging factors related to conducting regional nursing research. A 33-item survey based on the Iowa Model for Evidence-Based Practice was developed, validated, and determined to be reliable by the authors. Site coordinators organized and managed the orientation, administration, and collection of data from the 2,736 registered nurses who worked in 6 hospitals, 65 affiliated clinics, and 3 business units. Narrative notes taken by study investigators were analyzed for themes to determine challenging and facilitating factors for conducting regional research. Education and job title significantly predicted knowledge and ability to perform research activities but was not related to willingness to engage in research activities. Several environmental factors were associated with knowledge of, willingness to engage in, and ability to perform research utilization activities. Challenging and facilitating factors to conducting regional research were identified. Our research environment is changing to value research as shown in the philosophy, conceptual framework, and bylaws for the professional nursing staff. Novice-to-expert research utilization expectations are included in the promotional model for nursing. All RN job descriptions and the annual performance tool were revised to include responsibilities related to research activities. The Iowa Model for Evidence-Based Practice was adopted as the method for creating practice validation and change. Train-the-trainer educational and experiential sessions are being designed for nurse leaders; all new RN employees complete a self-assessment tool of research utilization knowledge and the nursing division strategic goals incorporate research utilization expectations. The elements of this plan may be useful for nurse executives. Healthcare systems are restructuring throughout the world and within the United States. These changes are occurring to better meet the evolving healthcare needs of the population through cost-effective approaches. Within the United States, emerging organized healthcare systems require research related to patient care outcomes and the health systems that can best address them.


Consumers' responses to front-of-pack labels that vary by interpretive content.
Previous research has shown that front-of-pack labels (FoPLs) can assist people to make healthier food choices if they are easy to understand and people are motivated to use them. There is some evidence that FoPLs providing an assessment of a food's health value (evaluative FoPLs) are easier to use than those providing only numerical information on nutrients (reductive FoPLs). Recently, a new evaluative FoPL (the Health Star Rating (HSR)) has been introduced to Australia and New Zealand. The HSR features a summary indicator, differentiating it from many other FoPLs being used around the world. The aim of this study was to understand how consumers of all ages use and make sense of reductive FoPLs and evaluative FoPLs including evaluative FoPLs with and without summary indicators. Ten focus groups were conducted in Perth, Western Australia with adults (n = 50) and children aged 10-17 years (n = 35) to explore reactions to one reductive FoPL (the Daily Intake Guide), an existing evaluative FoPL (multiple traffic lights), and a new evaluative FoPL (the HSR). Participants preferred the evaluative FoPLs over the reductive FoPL, with the strongest preference being for the FoPL with the summary indicator (HSR). Discussions revealed the cognitive strategies used when interpreting each FoPL (e.g., using cut offs, heuristics, and the process of elimination), which differed according to FoPL format. Most participants reported being motivated to use the evaluative FoPLs (particularly the HSR) to make choices about foods consumed as part of regular daily meals, but not for discretionary foods consumed as snacks or deserts. The findings provide further evidence of the potential utility of evaluative FoPLs in supporting healthy food choices and can assist policy makers in selecting between alternative FoPL formats.


A hierarchy of ATP-consuming processes in mammalian cells.
The rates of different ATP-consuming reactions were measured in concanavalin A-stimulated thymocytes, a model system in which more than 80% of the ATP consumption can be accounted for. There was a clear hierarchy of the responses of different energy-consuming reactions to changes in energy supply: pathways of macromolecule biosynthesis (protein synthesis and RNA/DNA synthesis) were most sensitive to energy supply, followed by sodium cycling and then calcium cycling across the plasma membrane. Mitochondrial proton leak was the least sensitive to energy supply. Control analysis was used to quantify the relative control over ATP production exerted by the individual groups of ATP-consuming reactions. Control was widely shared; no block of reactions had more than one-third of the control. A fuller control analysis showed that there appeared to be a hierarchy of control over the flux through ATP: protein synthesis > RNA/DNA synthesis and substrate oxidation > Na+ cycling and Ca2+ cycling > other ATP consumers and mitochondrial proton leak. Control analysis also indicated that there was significant control over the rates of individual ATP consumers by energy supply. Each ATP consumer had strong control over its own rate but very little control over the rates of the other ATP consumers.


Experimental production of EVI antibodies.
The production of an antibody which reacts with endocardium, blood vessels, and cardiac interstitial tissue (EVI factor) was experimentally obtained in rabbits immunized either with heterologous heart crude antigen, Trypanosoma cruzi antigen, a mixture of heart and T. cruzi antigens, kidney antigen or lung antigen, and complete Freund's adjuvant. Immunization lasted 2 months, with multiple subcutaneous and foot pad injections being made every 15th day. Preimmunization serum from each animal was used as normal control serum. Rabbit sera were tested by indirect immunofluorescence against mouse heart and skeletal muscle unfixed cryostat sections and revealed by fluoresceinated anti-rabbit globulin diluted 1:80. The highest titers appeared with anti-heart sera (1:192) and with anti-heart/T. cruzi sera (1:96). Incorporation of T. cruzi antigen in heart extract did not increase EVI antibody titers. It is suggested that the EVI factor is an antibody to many antigenic determinants, the majority of which are present in the heart and some of which cross-react with T. cruzi.


Moderate-to-Vigorous Physical Activity and Sedentary Behavior: Independent Associations With Body Composition Variables in Brazilian Children.
This study aimed to analyze the independent associations of accelerometer-determined sedentary behavior, physical activity, and steps/day with body composition variables in Brazilian children. 485 children wore accelerometers for 7 days. Variables included time in sedentary behavior and different physical activity intensities (light, moderate, vigorous, or moderate-to-vigorous) and steps/day. Body fat percentage was measured using a bioelectrical impedance scale, and BMI was calculated. Children spent 55.7% of the awake portion of the day in sedentary behavior, 37.6% in light physical activity, 4.6% in moderate physical activity, and 1.9% in vigorous physical activity. Moderate-to-vigorous physical activity and steps/day were negatively associated with body composition (BMI and body fat percentage) variables, independent of sex and sedentary behavior. Beta values were higher for vigorous physical activity than moderate physical activity. Vigorous physical activity was negatively associated with BMI (β-.1425) and body fat percentage (β-.3082; p < .0001). In boys, there were significant negative associations between moderate, vigorous, and moderate-to-vigorous physical activity and steps/day with body composition, and in girls, there was only a negative association with vigorous physical activity, independent of sedentary behavior. Moderate-to-vigorous physical activity and steps/day (in boys), but especially vigorous physical activity (in boys and girls), are associated with body composition, independent of sedentary behavior. Sedentary behavior was not related with any of the body composition variables once adjusted for moderate-to-vigorous physical activity.


The effectiveness of community-based interventions to improve maternal and infant health in the Northeast of Brazil.
To evaluate the effectiveness of a community-based intervention project aimed at reducing maternal and infant mortality in a poor urban district in the city of Natal, in the Northeast of Brazil. The intervention, called the ProNatal project, introduced a program of integrated community health care to a geographically defined population. The interventions included the establishment of antenatal clinics at the district's health centers, the opening of the maternity facilities at the polyclinic for low-risk deliveries, the introduction of a family planning clinic and a breast-feeding clinic, support from pediatricians for under-5 (well-baby) clinics, children's outpatient services and children's emergency care, and the introduction of health agents recruited from the local community. Representative surveys of the population were taken at the project's inception (July 1995) and then 30 months later (December 1997), using a general health questionnaire adapted to the local conditions. Mortality data were collected from local registration systems as well as from an autopsy survey of perinatal and infant deaths. During 1995 there were 4 maternal deaths from 1 195 pregnancies (maternal mortality of 335/100 000); three of the deaths were related to hypertension and one to uterine perforation after an illegal abortion. During 1998 (post-intervention), there were no maternal deaths in pregnancy or childbirth. In 1993 no deliveries took place at the polyclinic, but in 1998 there were 946 deliveries at the clinic without any serious complications. The method of delivery, the incidence of prematurity, and the incidence of low birthweight did not change significantly over the study period. In the post-intervention survey, 75% of women reported receiving contraceptive advice from a doctor in the preceding year, compared to 50% in the first sample. A mortality survey carried out in 1993-1995 estimated the infant mortality rate to be 60/1 000 live births. By 1998, using data collected locally by active surveillance, the infant mortality rate was 37/1 000 live births. The causes of infant death in both those periods were dominated by respiratory infections and diarrheal disease. Over 95% of both samples initiated breast-feeding, but a higher proportion of the post-intervention sample reported breast-feeding for longer than 6 months (41% vs. 32%, P = 0.0005). No differences were apparent in the use of under-5 clinics, but immunization rates improved. Post-intervention, significant improvements were documented in the mothers' understanding of basic hygiene, their knowledge of causes of common diseases, and their management of acute respiratory infections and diarrhea in children. This was particularly true for the households visited by a community health agent. Inequalities in health care in poor urban populations can be reduced by integrated community-based interventions, including the use of health agents recruited from the local community.


Total scattering reveals the hidden stacking disorder in a 2D covalent organic framework.
Interactions between extended π-systems are often invoked as the main driving force for stacking and crystallization of 2D organic polymers. In covalent organic frameworks (COFs), the stacking strongly influences properties such as the accessibility of functional sites, pore geometry, and surface states, but the exact nature of the interlayer interactions is mostly elusive. The stacking mode is often identified as eclipsed based on observed high symmetry diffraction patterns. However, as pointed out by various studies, the energetics of eclipsed stacking are not favorable and offset stacking is preferred. This work presents lower and higher apparent symmetry modifications of the imine-linked TTI-COF prepared through high- and low-temperature reactions. Through local structure investigation by pair distribution function analysis and simulations of stacking disorder, we observe random local layer offsets in the low temperature modification. We show that while stacking disorder can be easily overlooked due to the apparent crystallographic symmetry of these materials, total scattering methods can help clarify this information and suggest that defective local structures could be much more prevalent in COFs than previously thought. A detailed analysis of the local structure helps to improve the search for and design of highly porous tailor-made materials.


Serine racemase is associated with schizophrenia susceptibility in humans and in a mouse model.
Abnormal N-methyl-d-aspartate receptor (NMDAR) function has been implicated in the pathophysiology of schizophrenia. d-serine is an important NMDAR modulator, and to elucidate the role of the d-serine synthesis enzyme serine racemase (Srr) in schizophrenia, we identified and characterized mice with an ENU-induced mutation that results in a complete loss of Srr activity and dramatically reduced d-serine levels. Mutant mice displayed behaviors relevant to schizophrenia, including impairments in prepulse inhibition, sociability and spatial discrimination. Behavioral deficits were exacerbated by an NMDAR antagonist and ameliorated by d-serine or the atypical antipsychotic clozapine. Expression profiling revealed that the Srr mutation influenced several genes that have been linked to schizophrenia and cognitive ability. Transcript levels altered by the Srr mutation were also normalized by d-serine or clozapine treatment. Furthermore, analysis of SRR genetic variants in humans identified a robust association with schizophrenia. This study demonstrates that aberrant Srr function and diminished d-serine may contribute to schizophrenia pathogenesis.


National and state policies influencing the care of children affected by AIDS.
The portrait of HIV-affected children and youth that emerges from this policy overview is still one of children obscured from view by the shadow of their parents' and siblings' illness and policies that only address the needs of HIV-infected individuals. In addition, the secrecy and stigma that still surround HIV and AIDS make it difficult for HIV-affected children and youth to benefit as fully as they might from policies and programs that provide more generic types of care and assistance. Our failure as a nation to better illuminate the plight of HIV-affected children and youth can only leave us with a generation of children who are at greater risk of psychiatric illness, involvement with the criminal justice system, substance abuse, and contracting HIV. To avoid these consequences, both public and private sectors must place the spotlight on the development of new policies and programs designed to specifically meet their needs. Because the solutions defy traditional disciplinary and administrative boundaries, we also need to become more skilled at interagency planning and collaboration. No one system alone can be responsive to the many social, mental health, legal, and support needs of these children and their caretakers. More specifically, recommendations for improved systems of care to HIV-affected children, youth, and their families are as follows: To promote and fund cross-disciplinary initiatives among agencies that administer child welfare services, income supports, AIDS care, and children's mental health services at the national, state, and local levels to specifically meet the mental health, psychosocial, and permanency planning needs of HIV-affected children and youth. To provide training opportunities for Ryan White Title I, II, and III case managers on assessing the needs of HIV-affected children and youth, developmental theories and concepts, principles of family-centered care, and child welfare issues. To increase funding of the Ryan White CARE Act to permit a more family-centered approach to care across Titles. To establish clearer guidelines for establishing a proportional basis for funding services to infants, children, and women under the Ryan White CARE Act. To support legislative, educational, and advocacy efforts to make managed care and welfare reform more responsive to the needs of HIV-affected children, youth, and their families. To encourage states to provide additional funding for mental health services specifically targeted to HIV-affected children and youth that are sufficiently flexible and of a long enough duration to adequately meet their needs. Surely, as we move into a new millennium with the capacity to map the human genome and clearly view distant galaxies, we should be able to marshal the will and resources necessary to formulate a sufficiently focused effort to respond compassionately and effectively to the needs of a generation of AIDS-affected children and youth.


Development and validation of the Australian Midwifery Standards Assessment Tool (AMSAT) to the Australian Midwife Standards for Practice 2018.
The Australian Midwifery Standards Assessment Tool (AMSAT) was developed against the Competency Standards for the Midwife in 2017 to enable consistent assessment of midwifery student performance in practice-based settings. The AMSAT requires revision and re-validation as the competency standards have now been superseded by the Midwife Standards for Practice 2018. This research revised and validated the AMSAT to assess performance of midwifery students against the Midwife Standards for Practice 2018 and assessed its sensitivity. A mixed-methods approach was used in a two-phase process. Phase one involved the re-wording of the AMSAT and behavioural cue statements in an iterative participatory process with midwifery academics, assessors and students. The tool was field-tested in different assessment environments in phase two. Completed assessment forms were statistically analyzed, whilst assessor surveys were analysed using descriptive statistics and qualitative content analysis. Analysis of AMSAT (n=255) indicates the tool as: internally reliable (Cronbach alpha>.9); valid (eigenvalue of 16.6 explaining 67% of variance); and sensitive (score analysis indicating increased levels of proficiency with progressive student experience). Analysis of surveys (n=108) found acceptance of the tool for the purpose of summative and formative assessment, and in the provision of feedback to midwifery students on their performance. This study demonstrates that the re-developed AMSAT is a valid, reliable and acceptable tool to assess midwifery students' performance against the Australian Midwife Standards for Practice This user-friendly tool can be used to standardize midwifery student assessment in Australia and enable continued benchmarking across education programs.


Mental Work Demands and Late-Life Cognitive Impairment: Results From the Shanghai Aging Study.
The aim of this study is to explore the association between mental work demands (MWDs) and late-life cognition among Chinese older adults. Data were drawn from the baseline of the Shanghai Aging Study. Among 3,546 participants, the mean MWDs index of participants with dementia, mild cognitive impairment (MCI), and cognitive normal were 2.8 ( SD = 0.5), 2.8 ( SD = 0.5), and 2.9 ( SD = 0.4), respectively. Mini Mental State Examination (MMSE) scores in group with low MWDs level decreased more dramatically by increasing age than that in groups with middle and high MWDs level ( p < .001). In a subgroup of participants with education ≥6 years, we found a positive correlation between MWDs index and MMSE score after adjusting for gender, age, body mass index, income, living alone, smoking, drinking, anxiety, depression, heart disease, hypertension, diabetes, and APOE-ε4 (apolipoprotein), by generalized linear model analysis ( p < .001). Our results suggest that long-time mental work stimulation during adulthood would have potential benefits to late-life cognition.


Updating the Genome of the Elite Rice Variety Kongyu131 to Expand Its Ecological Adaptation Region.
As an elite rice variety cultivated in the third accumulative temperature belt in Heilongjiang province, China, Kongyu131 has many excellent traits, such as high quality, high stability, early maturation and cold resistance. However, as with other crop varieties, Kongyu131 has regional restrictions, exhibiting decreased yields when grown at low latitudes. To address these problems, two populations were constructed from cross between japonica and indica varieties. QTL analyses were performed with these two populations to detect regional adaptation related quantitative trait locus. Results in a BC1F6 backcross inbred line population with 168 lines derived from cross between Kongyu131 and GKMP showed a large pleiotropic QTL near 9 Mb on chromosome 7, which significantly delayed the HD of Kongyu131 and increased the plant height (PH), length of main panicle (LMP), number of primary branches (NPB) and grain number of main panicles (GNP). We also found a similar QTL in the population BC3F2 derived from Kongyu131 and GKLPL. Based on the QTL, we developed a gene module named mRA7 with 5 single-nucleotide polymorphism (SNP) markers around the QTL. Through a foreground and background selection based on 197 SNP markers evenly distributed over the 12 chromosomes, we obtained a new plant (a single point substitution line, SPSL) with a new Kongyu131 genome, carrying only a small chromosomal fragment less than 800 kb from GKLPL. The background recovery ratio of the SPSL was 99.8%. Compared with Kongyu131, the SPSL exhibited a significant HD delay of approximately 31 days and increased PH, LMP and GNP values when planted in Heilongjiang province. When cultivated in Guangdong province, HD of SPSL showed only 16 days delay, and less increase in PH, LMP and GNP than in Heilongjiang province. Phenotypic evaluation showed that the SPSL could be moved to south by more than 3 latitude units and cultivated in low-latitude regions. This study exemplifies the feasibility of expanding the regions of cultivation of elite rice varieties via similar methods.


A prospective study of microalbuminuria and incident coronary heart disease and its prognostic significance in a British population: the EPIC-Norfolk study.
Microalbuminuria is associated with an increased risk of cardiovascular and renal disease in patients with diabetes and hypertension. The role of microalbuminuria as a predictor of coronary heart disease (CHD) has not been examined in large general-population cohorts, and its prognostic significance in persons with established CHD is uncertain. The authors examined the relation between microalbuminuria and incident CHD (1993-2002) in a population-based British cohort of 22,368 men and women aged 40-79 years without prevalent baseline CHD and evaluated its prognostic significance in 1,596 participants with baseline CHD. Participants were members of the Norfolk, United Kingdom, component of the European Prospective Investigation into Cancer and Nutrition (the EPIC-Norfolk Study). At baseline, participants were categorized into normoalbuminuria, microalbuminuria, and macroalbuminuria groups. During an average of 6.4 years of follow-up, 800 primary CHD events were registered. The age-adjusted incidence of CHD increased significantly across ordered categories of albuminuria (4.3, 4.4, and 5.6/1,000 person-years across tertiles of normoalbuminuria, 7.1/1,000 person-years for microalbuminuria, and 12.2/1,000 person-years for macroalbuminuria; p for trend < 0.001). The multivariate hazard ratio for incident primary CHD was 1.36 (95% confidence interval (CI): 1.12, 1.64) for microalbuminuria and 1.59 (95% CI: 1.10, 2.37) for macroalbuminuria. Among participants with established baseline CHD, the independent risk of all-cause mortality associated with microalbuminuria was 1.61 (95% CI: 1.19, 2.07). Microalbuminuria may be useful in identifying persons at increased risk of CHD and subsequent death in the general population.


News or social media? Socio-economic divide of mobile service consumption.
Reliable and timely information on socio-economic status and divides is critical to social and economic research and policing. Novel data sources from mobile communication platforms have enabled new cost-effective approaches and models to investigate social disparity, but their lack of interpretability, accuracy or scale has limited their relevance to date. We investigate the divide in digital mobile service usage with a large dataset of 3.7 billion time-stamped and geo-referenced mobile traffic records in a major European country, and find profound geographical unevenness in mobile service usage-especially on news, e-mail, social media consumption and audio/video streaming. We relate such diversity with income, educational attainment and inequality, and reveal how low-income or low-education areas are more likely to engage in video streaming or social media and less in news consumption, information searching, e-mail or audio streaming. The digital usage gap is so large that we can accurately infer the socio-economic status of a small area or even its Gini coefficient only from aggregated data traffic. Our results make the case for an inexpensive, privacy-preserving, real-time and scalable way to understand the digital usage divide and, in turn, poverty, unemployment or economic growth in our societies through mobile phone data.


Purification and characterization of a two-component monooxygenase that hydroxylates nitrilotriacetate from "Chelatobacter" strain ATCC 29600.
An assay based on the consumption of nitrilotriacetate (NTA) was developed to measure the activity of NTA monooxygenase (NTA-Mo) in cell extracts of "Chelatobacter" strain ATCC 29600 and to purify a functional, NTA-hydroxylating enzyme complex. The complex consisted of two components that easily dissociated during purification and upon dilution. Both components were purified to more than 95% homogeneity, and it was possible to reconstitute the functional, NTA-hydroxylating enzyme complex from pure component A (cA) and component B (cB). cB exhibited NTA-stimulated NADH oxidation but was unable to hydroxylate NTA. It had a native molecular mass of 88 kDa and contained flavin mononucleotide (FMN). cA had a native molecular mass of 99 kDa. No catalytic activity has yet been shown for cA alone. Under unfavorable conditions, NADH oxidation was partly or completely uncoupled from hydroxylation, resulting in the formation of H2O2. Optimum hydroxylating activity was found to be dependent on the molar ratio of the two components, the absolute concentration of the enzyme complex, and the presence of FMN. Uncoupling of the reaction was favored in the presence of high salt concentrations and in the presence of flavin adenine dinucleotide. The NTA-Mo complex was sensitive to sulfhydryl reagents, but inhibition was reversible by addition of excess dithiothreitol. The Km values for Mg(2+)-NTA, FMN, and NADH were determined as 0.5 mM, 1.3 microM, and 0.35 mM, respectively. Of 26 tested compounds, NTA was the only substrate for NTA-Mo.


Six git genes encode a glucose-induced adenylate cyclase activation pathway in the fission yeast Schizosaccharomyces pombe.
An important eukaryotic signal transduction pathway involves the regulation of the effector enzyme adenylate cyclase, which produces the second messenger, cAMP. Previous genetic analyses demonstrated that glucose repression of transcription of the Schizosaccharomyces pombe fbp1 gene requires the function of adenylate cyclase, encoded by the git2 gene. As mutations in git2 and in six additional git genes are suppressed by exogenous cAMP, these 'upstream' git genes were proposed to act to produce a glucose-induced cAMP signal. We report here that assays of cAMP levels in wild-type and various mutant S. pombe cells, before and after exposure to glucose, show that this is the case. The data suggest that the cAMP signal results from the activation of adenylate cyclase. Therefore these 'upstream' git genes appear to encode a glucose-induced adenylate cyclase activation pathway. Assays of cAMP on a strain carrying a mutation in the git6 gene, which acts downstream of adenylate cyclase, indicate that git6 may function to feedback regulate adenylate cyclase activity. Thus git6 may encode a cAMP-dependent protein kinase.


Enzymes of ethylene biosynthesis.
The properties of enzymes involved in ethylene biosynthesis are reviewed and progress toward the purification of these enzymes is described. The enzyme whose activity usually limits ethylene biosynthesis is 1-aminocyclopropane-1-carboxylate (ACC) synthase. Even though its level in plants is extremely low, it has now been purified from several sources. The enzyme that converts ACC to ethylene does not survive homogenization, apparently because it is membrane-bound and because its activity requires membrane integrity. Properties of this enzyme have been elucidated in vivo and in vacuolar preparations which possess the capacity to convert ACC to ethylene.


Control by fibroblast growth factor of differentiation in the BC3H1 muscle cell line.
The regulation of creatine phosphokinase (CPK) expression by polypeptide growth factors has been examined in the clonal mouse muscle BC3H1 cell line. After arrest of cell growth by exposure to low concentrations of serum, BC3H1 cells accumulate high levels of muscle-specific proteins including CPK. The induction of this enzyme is reversible in the presence of high concentrations of fetal calf serum, which cause quiescent, differentiated cells to reenter the cell cycle. Under these conditions, the rate of CPK synthesis is drastically reduced. We show in the present communication that either pituitary-derived fibroblast growth factor (FGF) or brain-derived FGF are as effective as serum in repressing the synthesis of CPK when added to quiescent, differentiated cells. The decrease in the rate of synthesis of CPK occurs within 22 h after the addition of pituitary FGF to the cells. Pituitary FGF had very little effect, if any, on the rate CPK degradation. The overall rate of protein synthesis and the pattern of synthesis of the major polypeptides made by these cells was not altered by the addition of FGF. Although pituitary FGF was mitogenic for BC3H1 cells, the rate of cell growth was not absolutely correlated with the extent of repression of CPK. Brain-derived FGF fully repressed CPK induction under conditions where it showed no significant mitogenic activity. These results show that the expression of a muscle-specific protein, CPK, can be controlled by a single defined polypeptide growth factor in fully differentiated cultures, and that initiation of cell division is not required for their regulation to take place.


Arterial spin-labeling MRI can identify the presence and intensity of collateral perfusion in patients with moyamoya disease.
Determining the presence and adequacy of collateral blood flow is important in cerebrovascular disease. Therefore, we explored whether a noninvasive imaging modality, arterial spin labeling (ASL) MRI, could be used to detect the presence and intensity of collateral flow using digital subtraction angiography (DSA) and stable xenon CT cerebral blood flow as gold standards for collaterals and cerebral blood flow, respectively. ASL and DSA were obtained within 4 days of each other in 18 patients with Moyamoya disease. Two neurointerventionalists scored DSA images using a collateral grading scale in regions of interest corresponding to ASPECTS methodology. Two neuroradiologists similarly scored ASL images based on the presence of arterial transit artifact. Agreement of ASL and DSA consensus scores was determined, including kappa statistics. In 15 patients, additional quantitative xenon CT cerebral blood flow measurements were performed and compared with collateral grades. The agreement between ASL and DSA consensus readings was moderate to strong, with a weighted kappa value of 0.58 (95% confidence interval, 0.52-0.64), but there was better agreement between readers for ASL compared with DSA. Sensitivity and specificity for identifying collaterals with ASL were 0.83 (95% confidence interval, 0.77-0.88) and 0.82 (95% confidence interval, 0.76-0.87), respectively. Xenon CT cerebral blood flow increased with increasing DSA and ASL collateral grade (P<0.05). ASL can noninvasively predict the presence and intensity of collateral flow in patients with Moyamoya disease using DSA as a gold standard. Further study of other cerebrovascular diseases, including acute ischemic stroke, is warranted.


Interventions for nausea and vomiting in early pregnancy.
Nausea and vomiting are the most common symptoms experienced in early pregnancy, with nausea affecting between 70 and 85% of women. About half of pregnant women experience vomiting. To assess the effects of different methods of treating nausea and vomiting in early pregnancy. We searched the Cochrane Pregnancy and Childbirth Group trials register (December 2002) and the Cochrane Central Register of Controlled Trials (The Cochrane Library, Issue 4, 2002). Randomised trials of any treatment for nausea and/or vomiting in early pregnancy. Two reviewers assessed the trial quality and extracted the data independently. Twenty-eight trials met the inclusion criteria. For milder degrees of nausea and vomiting, 21 trials were included. These trials were of variable quality. Nausea treatments were: different antihistamine medications, vitamin B6 (pyridoxine), the combination tablet Debendox (Bendectin), P6 acupressure and ginger. For hyperemesis gravidarum, seven trials were identified testing treatments with oral ginger root extract, oral or injected corticosteroids or injected adrenocorticotropic hormone (ACTH), intravenous diazepam and acupuncture. Based on 12 trials, there was an overall reduction in nausea from anti-emetic medication (odds ratio 0.16, 95% confidence interval 0.08 to 0.33). Anti-emetic medication appears to reduce the frequency of nausea in early pregnancy. There is some evidence of adverse effects, but there is very little information on effects on fetal outcomes from randomised controlled trials. Of newer treatments, pyridoxine (vitamin B6) appears to be more effective in reducing the severity of nausea. The results from trials of P6 acupressure are equivocal. No trials of treatments for hyperemesis gravidarum show any evidence of benefit. Evidence from observational studies suggests no evidence of teratogenicity from any of these treatments.


Resolving the phylogenetic position of Darwin's extinct ground sloth (Mylodon darwinii) using mitogenomic and nuclear exon data.
Mylodon darwinii is the extinct giant ground sloth named after Charles Darwin, who first collected its remains in South America. We have successfully obtained a high-quality mitochondrial genome at 99-fold coverage using an Illumina shotgun sequencing of a 12 880-year-old bone fragment from Mylodon Cave in Chile. Low level of DNA damage showed that this sample was exceptionally well preserved for an ancient subfossil, probably the result of the dry and cold conditions prevailing within the cave. Accordingly, taxonomic assessment of our shotgun metagenomic data showed a very high percentage of endogenous DNA with 22% of the assembled metagenomic contigs assigned to Xenarthra. Additionally, we enriched over 15 kb of sequence data from seven nuclear exons, using target sequence capture designed against a wide xenarthran dataset. Phylogenetic and dating analyses of the mitogenomic dataset including all extant species of xenarthrans and the assembled nuclear supermatrix unambiguously place Mylodon darwinii as the sister-group of modern two-fingered sloths, from which it diverged around 22 million years ago. These congruent results from both the mitochondrial and nuclear data support the diphyly of the two modern sloth lineages, implying the convergent evolution of their unique suspensory behaviour as an adaption to arboreality. Our results offer promising perspectives for whole-genome sequencing of this emblematic extinct taxon.


A digitally facilitated citizen-science driven approach accelerates participant recruitment and increases study population diversity.
Our aim was to assess whether a novel approach of digitally facilitated, citizen-science research, as followed by the Swiss Multiple Sclerosis Registry (Swiss MS Registry), leads to accelerated participant recruitment and more diverse study populations compared with traditional research studies where participants are mostly recruited in study centres without the use of digital technology. The Swiss MS Registry is a prospective, longitudinal, observational study covering all Switzerland. Participants actively contribute to the Swiss MS Registry, from defining research questions to providing data (online or on a paper form) and co-authoring papers. We compared the recruitment dynamics over the first 18 months with the a priori defined recruitment goals and assessed whether a priori defined groups were enrolled who are likely to be missed by traditional research studies. The goal to recruit 400 participants in the first year was reached after only 20 days, and by the end of 18 months 1700 participants had enrolled in the Swiss MS Registry, vastly exceeding expectations. Of the a priori defined groups with potential underrepresentation in other studies, 645 participants (46.5%) received care at a private neurology practice, 167 participants (12%) did not report any use of healthcare services in the past 12 months, 32 (2.3%) participants lived in rural mountainous areas, and 20 (2.0% of the 1041 for whom this information was available) lived in a long-term care facility. Having both online and paper options increased diversity of the study population in terms of geographic origin and type and severity of disease, as well as use of health care services. In particular, paper enrolees tended to be older, more frequently affected by progressive MS types and more likely to have accessed healthcare services in the past 12 months. Academic and industry-driven medical research faces substantial challenges in terms of patient involvement, recruitment, relevance and generalisability. Digital studies and stakeholder engagement may have enormous potential for medical research. But many digital studies are based on limited participant information and/or informed consent and unclear data ownership, and are subject to selection bias, confounding and information bias. The Swiss MS Registry serves as an example of a digitally enhanced, citizen-science study that leverages the advantages of both traditional medical research, with its established research methods, and novel societal and technological developments, while mitigating their ethical and legal disadvantages and risks.