Criteria for validation and selection of cognitive tests for investigating the effects of foods and nutrients.
This review is an output of the International Life Sciences Institute (ILSI) Europe Marker Initiative, which aims to identify evidence-based criteria for selecting adequate measures of nutrient effects on health through comprehensive literature review. Experts in cognitive and nutrition sciences examined the applicability of these proposed criteria to the field of cognition with respect to the various cognitive domains usually assessed to reflect brain or neurological function. This review covers cognitive domains important in the assessment of neuronal integrity and function, commonly used tests and their state of validation, and the application of the measures to studies of nutrition and nutritional intervention trials. The aim is to identify domain-specific cognitive tests that are sensitive to nutrient interventions and from which guidance can be provided to aid the application of selection criteria for choosing the most suitable tests for proposed nutritional intervention studies using cognitive outcomes. The material in this review serves as a background and guidance document for nutritionists, neuropsychologists, psychiatrists, and neurologists interested in assessing mental health in terms of cognitive test performance and for scientists intending to test the effects of food or food components on cognitive function.


Multitechnology Biofabrication: A New Approach for the Manufacturing of Functional Tissue Structures?
Most available 3D biofabrication technologies rely on single-component deposition methods, such as inkjet, extrusion, or light-assisted printing. It is unlikely that any of these technologies used individually would be able to replicate the complexity and functionality of living tissues. Recently, new biofabrication approaches have emerged that integrate multiple manufacturing technologies into a single biofabrication platform. This has led to fabricated structures with improved functionality. In this review, we provide a comprehensive overview of recent advances in the integration of different manufacturing technologies with the aim to fabricate more functional tissue structures. We provide our vision on the future of additive manufacturing (AM) technology, digital design, and the use of artificial intelligence (AI) in the field of biofabrication.


The functional unit of the renal type IIa Na+/Pi cotransporter is a monomer.
The composition of the functional unit of the rat renal type IIa Na(+)/P(i) cotransporter (NaPi-IIa) was investigated by using two approaches based on the differential sensitivities of the wild type (WT) and mutant S460C proteins to 2-aminoethylmethanethiosulfonate hydrobromide (MTSEA), a charged cysteine modifier. Transport activity of S460C is completely blocked after incubation in MTSEA, whereas that of the WT remains unaffected. First, Xenopus laevis oocytes were coinjected with cRNAs coding for the WT and S460C in different proportions, and the transport inhibition after MTSEA incubation was assayed by electrophysiology. The relationship between MTSEA inhibition and proportion of cRNA was consistent with that for a functional monomer. Second, concatameric proteins were constructed that either comprised two WT proteins (WT-WT), two S460C mutants (S460C-S460C), or one of each (WT-S460C). Western blots of oocytes injected with fusion protein cRNA showed bands at approximately 200 kDa, whereas a main band at approximately 90 kDa was obtained for the WT cRNA alone. The kinetic properties of concatamers were the same as for the single proteins. Transport activity of the WT-WT concatamer was unaffected by MTSEA incubation, fully inhibited for S460C-S460C, but 50% inhibited for WT-S460C. This behavior was also consistent with NaPi-IIa being a functional monomer.


Skeletal Muscle Metabolic Dysfunction in Patients With Malignant Hyperthermia Susceptibility.
Malignant hyperthermia (MH), a pharmacogenetic disorder of skeletal muscle, presents with a potentially lethal hypermetabolic reaction to certain anesthetics. However, some MH-susceptible patients experience muscle weakness, fatigue, and exercise intolerance in the absence of anesthetic triggers. The objective of this exploratory study was to elucidate the pathophysiology of exercise intolerance in patients tested positive for MH with the caffeine-halothane contracture test. To this end, we used phosphorus magnetic resonance spectroscopy, blood oxygen level-dependent functional magnetic resonance imaging (MRI), and traditional exercise testing to compare skeletal muscle metabolism in MH-positive patients and healthy controls. Skeletal muscle metabolism was assessed using phosphorus magnetic resonance spectroscopy and blood oxygen level-dependent functional MRI in 29 MH-positive patients and 20 healthy controls. Traditional measures of physical capacity were employed to measure aerobic capacity, anaerobic capacity, and muscle strength. During 30- and 60-second exercise, MH-positive patients had significantly lower ATP production via the oxidative pathway compared to healthy controls. MH-positive patients also had a longer recovery time with blood oxygen level-dependent functional MRI compared to healthy controls. Exercise testing revealed lower aerobic and anaerobic capacity in MH-positive patients compared to healthy controls. Results of this exploratory study suggest that MH-positive patients have impaired aerobic metabolism compared to healthy individuals. This could explain the exercise intolerance exhibited in MH-susceptible patient population.


Bacterial protein degradation by different rumen protozoal groups.
Bacterial predation by protozoa has the most deleterious effect on the efficiency of N use within the rumen, but differences in activity among protozoal groups are not completely understood. Two in vitro experiments were conducted to identify the protozoal groups more closely related with rumen N metabolism. Rumen protozoa were harvested from cattle and 7 protozoal fractions were generated immediately after sampling by filtration through different nylon meshes at 39 °C, under a CO(2) atmosphere to maintain their activity. Protozoa were incubated with (14)C-labeled bacteria to determine their bacterial breakdown capacity, according to the amount of acid-soluble radioactivity released. Epidinium tended to codistribute with Isotricha and Entodinium with Dasytricha; therefore, their activity was calculated together. This study demonstrated that big Diplodiniinae had the greatest activity per cell (100 ng bacterial CP per protozoa and hour), followed by Epidinium plus Isotricha (36.4), small Diplodiniinae (34.2), and Entodinium plus Dasytricha (14.8), respectively. However, the activity per unit of protozoal volume seemed to vary, depending on the protozoal taxonomy. Small Diplodiniinae had the greatest activity per volume (325 ng bacterial CP per protozoal mm(3) and hour), followed by big Diplodiniinae (154), Entodinium plus Dasytricha (104), and Entodinium plus Dasytricha (25.6). A second experiment was conducted using rumen fluid from holotrich-monofaunated sheep. This showed that holotrich protozoa had a limited bacterial breakdown capacity per cell (Isotricha 9.44 and Dasytricha 5.81 ng bacterial CP per protozoa and hour) and per protozoal volume (5.97 and 76.9 ng bacterial CP per protozoal mm(3) and hour, respectively). Therefore, our findings indicated that a typical protozoal population (10(6) total protozoa/mL composed by Entodinium sp. 88%, Epidinium sp. 7%, and other species 4%) is able to break down ~17% of available rumen bacteria every hour. Entodinium sp. is responsible for most of this bacterial breakdown (70 to 75%), followed by Epidinium sp. (16 to 24%), big Diplodiniinae (4 to 6%), and small Diplodiniinae (2 to 6%), whereas holotrich protozoa have a negligible activity (Dasytricha sp. 0.6 to 1.2% and Isotricha sp. 0.2 to 0.5%). This in vitro information must be carefully interpreted, but it can be used to indicate which protozoal groups should be suppressed to improve microbial protein synthesis in vivo.


Progress in researches about focal adhesion kinase in gastrointestinal tract.
Focal adhesion kinase (FAK) is a 125-kDa non-receptor protein tyrosine. Growth factors or the clustering of integrins facilitate the rapid phosphorylation of FAK at Tyr-397 and this in turn recruits Src-family protein tyrosine kinases, resulting in the phosphorylation of Tyr-576 and Tyr-577 in the FAK activation loop and full catalytic FAK activation. FAK plays a critical role in the biological processes of normal and cancer cells including the gastrointestinal tract. FAK also plays an important role in the restitution, cell survival and apoptosis and carcinogenesis of the gastrointestinal tract. FAK is over-expressed in cancer cells and its over-expression and elevated activities are associated with motility and invasion of cancer cells. FAK has been proposed as a potential target in cancer therapy. Small molecule inhibitors effectively inhibit the kinase activity of FAK and show a potent inhibitory effect for the proliferation and migration of tumor cells, indicating a high potential for application in cancer therapy.


CD46-mediated transduction of a species D adenovirus vaccine improves mucosal vaccine efficacy.
The high levels of preexisting immunity against Adenovirus type 5 (Ad5) have deemed Ad5 unusable for translation as a human vaccine vector. Low seroprevalent alternative viral vectors may be less impacted by preexisting immunity, but they may also have significantly different phenotypes from that of Ad5. In this study we compare species D Ads (26, 28, and 48) to the species C Ad5. In vitro transduction studies show striking differences between the species C and D viruses. Most notably, Ad26 transduced human dendritic cells much more effectively than Ad5. In vivo imaging studies showed strikingly different transgene expression profiles. The Ad5 virus was superior to the species D viruses in BALB/c mice when delivered intramuscularly. However, the inverse was true when the viruses were delivered mucosally via the intranasal epithelia. Intramuscular transduction was restored in mice that ubiquitously expressed human CD46, the primary receptor for species D viruses. We analyzed both species C and D Ads for their ability to induce prophylactic immunity against influenza in the CD46 transgenic mouse model. Surprisingly, the species D vaccines again failed to induce greater levels of protective immunity as compared with the species C Ad5 when delivered intramuscularly. However, the species D Ad vaccine vector, Ad48, induced significantly greater protection as compared with Ad5 when delivered mucosally via the intranasal route in CD46 transgenic mice. These data shed light on the complexities between the species and types of Ad. Our findings indicate that more research will be required to identify the mechanisms that play a key role in the induction of protective immunity induced by species D Ad vaccines.


Improving child and parenting outcomes following paediatric acquired brain injury: a randomised controlled trial of Stepping Stones Triple P plus Acceptance and Commitment Therapy.
Persistent behavioural difficulties are common following paediatric acquired brain injury (ABI). Parents and families also experience heightened stress, psychological symptoms and burden, and there is evidence of a reciprocal relationship between parent and child functioning, which may be mediated by the adoption of maladaptive parenting practices. Despite this, there is currently a paucity of research in family interventions in this population. The aim of this study was to determine the efficacy of Stepping Stones Triple P: Positive Parenting Program (SSTP), with an Acceptance and Commitment Therapy (ACT) workshop, in improving child outcomes and parenting practices following paediatric ABI. Fifty-nine parents of children (mean age 7 years, SD 3 years, 1 month; 35 males, 24 females) with ABI (Traumatic injuries 58%, Tumour 17%, Encephalitis or meningitis 15%, Cardiovascular accident 7%, Hypoxia 3%) who were evidencing at least mild behaviour problems were randomly assigned to treatment or care-as-usual conditions over 10 weeks. Mixed-model repeated-measures linear regression analyses were conducted to compare conditions from pre- to postintervention on child behavioural and emotional functioning (Eyberg Child Behavior Inventory, Strengths and Difficulties Questionnaire) and dysfunctional parenting style (Parenting Scale). Assessment of maintenance of change was conducted at a 6-month follow-up. The trial was registered on Australian New Zealand Clinical Trials Registry (ID: ACTRN12610001051033, www.anzctr.org.au). Significant time-by-condition interactions were identified on number and intensity of child behaviour problems, child emotional symptoms and parenting laxness and overreactivity, indicating significant improvements in the treatment condition, with medium-to-large effect sizes. Most improvements were maintained at 6 months. Group parenting interventions incorporating Triple P and ACT may be efficacious in improving child and parenting outcomes following paediatric ABI.


Detection of in situ mammary cancer in a transgenic mouse model: in vitro and in vivo MRI studies demonstrate histopathologic correlation.
Improving the prevention and detection of preinvasive ductal carcinoma in situ (DCIS) is expected to lower both morbidity and mortality from breast cancer. Transgenic mouse models can be used as a 'test bed' to develop new imaging methods and to evaluate the efficacy of candidate preventive therapies. We hypothesized that despite its microscopic size, early murine mammary cancer, including DCIS, might be accurately detected by MRI. C3(1) SV40 TAg female mice (n=23) between 10 and 18 weeks of age were selected for study. Eleven mice were subjected to in vitro imaging using a T(2)-weighted spin echo sequence and 12 mice were selected for in vivo imaging using a T(1)-weighted gradient echo, a T(2)-weighted spin echo and high spectral and spatial resolution imaging sequences. The imaged glands were carefully dissected, formalin fixed and paraffin embedded, and then H&E stained sections were obtained. The ratio of image-detected versus histologically detected cancers was obtained by reviewing the MR images and H&E sections independently and using histology as the gold standard. MR images were able to detect 12/12 intramammary lymph nodes, 1/1 relatively large (approximately 5 mm) tumor, 17/18 small (approximately 1 mm) tumors and 13/16 ducts distended with DCIS greater than 300 microm. Significantly, there were no false positives--i.e., image detection always corresponded to a histologically detectable cancer in this model. These results indicate that MR imaging can reliably detect both preinvasive in situ and early invasive mammary cancers in mice with high sensitivity. This technology is an important step toward the more effective use of non-invasive imaging in pre-clinical studies of breast cancer prevention, detection and treatment.


Social networks of PLHA in Uganda: implications for mobilizing PLHA as agents for prevention.
The objective of this study was to describe the social networks of people living with HIV/AIDS (PLHA) and explore the implications social network characteristics might have for mobilizing PLHA as prevention agents. Thirty-nine PLHA attending an HIV clinic in Kampala, Uganda provided information on themselves and on 20 network members. Based on these data, descriptive statistics for social network composition and structure were calculated. Research questions relating these network characteristics to treatment-related variables such as time since diagnosis, ART status, and time on ART were investigated. Analyses reveal that, in general, network members know the PLHA's status, are trusted, provide advice and support, and are well-connected to each other. Network features (e.g., proportion of individuals who know the PLHA's status) are related to the previously mentioned treatment variables. Findings suggest that PLHA surround themselves with a social context that enables PLHA to feel fairly protected and supported if they choose to discuss HIV and prevention. With respect to treatment, those on ART may be better prepared to act as prevention advocates.


A temporal study of the expression of the capsid, cytoplasmic inclusion and nuclear inclusion proteins of tobacco etch potyvirus in infected plants.
Young leaves of tobacco, systemically infected by tobacco etch potyvirus (TEV), were examined for the presence and distribution of four virus encoded proteins [capsid, cytoplasmic inclusion (CI) and two nuclear inclusion (NI) proteins] at various time periods after inoculation of expanded leaves of the plants. The analyses were carried out by ELISA and by immunogold electron microscopy of thin sections of the leaves. All four proteins were detected simultaneously in the systemic leaves for the first time on the fifth day after inoculation of the expanded leaves. All four proteins increased in concentration until the seventh day and then showed no further increase with the exception of the capsid protein which continued to accumulate. The CI protein was first detected in association with the plasmalemma/cell wall and was subsequently found mostly in the form of pinwheels in the cytoplasm. The two NI proteins were found at all times after infection within the nucleus, although small concentrations were detected in the cytoplasm. These experiments suggest that both the NIa and NIb proteins are transported into the nucleus immediately after synthesis. At the earliest time periods after infection, high concentrations of these proteins (NIa and NIb) were found in their non-inclusion form in the nucleolus. At 14 days after infection, both proteins were found only as inclusions in the nucleus. The capsid protein was found at all stages of infection only in the cytoplasm.


Abortion training at multiple sites: an unexpected curriculum for teaching systems-based practice.
In 1999, the Accreditation Council for Graduate Medical Education endorsed systems-based practice as one of six general competencies. The objective is to explore the paradigm of teaching residents systems-based practice during a women's health rotation that included abortion training in multiple settings. During a routine women's health rotation, residents from two urban family medicine residency programs received early abortion training at a high-volume abortion clinic and their continuity clinic. Thirty-min semistructured interviews were conducted with all 26 residents who rotated between July 2005 and August 2006. Transcripts were analyzed using thematic codes. Through exposure to different healthcare delivery systems, residents learned about systems-based practice, including understanding the failure of the larger system to meet patients' reproductive healthcare needs, differences between two systems, and potential systems barriers they might face as providers. Abortion training in multiple settings may serve as a paradigm for teaching systems-based practice during other rotations that include training in multiple sites.


European Psychiatric Association (EPA) guidance on prevention of mental disorders.
There is considerable evidence that various psychiatric conditions can be prevented through the implementation of effective evidence-based interventions. Since a large proportion of lifetime mental illness starts before adulthood, such interventions are particularly important during childhood and adolescence. Prevention is important for the sustainable reduction of the burden of mental disorder since once it has arisen, treatment can only reduce a relatively small proportion of such burden. The challenge for clinicians is to incorporate such interventions into non-clinical and clinical practice as well as engaging with a range of other service providers including public health. Similar strategies can be employed in both the European and global contexts. Promotion of mental well-being can prevent mental disorder but is also important in the recovery from mental disorder. This guidance should be read in conjunction with the EPA Guidance on Mental Health Promotion. This guidance draws on preparatory work for the development of England policy on prevention of mental disorder which used a wide range of sources.


External versus endoscopic dacryocystorhinostomy for acquired nasolacrimal duct obstruction in a tertiary referral center.
To compare success rates of external dacryocystorhinostomy (DCR) and endoscopic endonasal DCR for acquired nasolacrimal duct obstruction (NLDO). Retrospective, comparative, nonrandomized clinical study. One hundred forty-three patients (176 surgeries) operated for acquired NLDO. A review of electronic medical records of patients with acquired NLDO who underwent DCR at the Jules Stein Eye Institute from 1999 to 2004 was performed. Data regarding the lacrimal drainage system, comprehensive eye examination, surgical outcome, and postoperative nasal endoscopy were analyzed. Surgery failure was defined as (1) no marked improvement in tearing or any episode of postoperative dacryocystitis, (2) inability to irrigate the lacrimal system postoperatively, and (3) postoperative nasal endoscopy with scarring in the intranasal osteotomy or no visualization of fluorescein dye. Postoperative nasal endoscopy was performed in all failed cases and in >50% of all patients. One hundred forty-three patients (48 male and 95 female; mean age, 63 years) underwent 176 DCR surgeries for acquired NLDO. Success was achieved in 135 cases (76.7%), and failure in 41 (23.3%). Of the 41 failed cases, anatomical obstruction at the fistula site was found in 20 (49% of failed cases), whereas functional failure with no evidence of obstruction was found in 21 (51%). Surgery revision was performed in 22 cases (12.5%), but it was successful in only 9 (5.1%); patients who failed the first revision were likely to fail additional revisions (P = 0.02). History of facial trauma was associated with surgery failure. In our patients, endoscopic DCR (86 cases) had a significantly higher success rate than external DCR (90 cases), 84% versus 70% (P = 0.03). Complications included 1 patient with nose bleeding on the first postoperative day that resolved with nasal packing and 2 patients with sump syndrome that resolved after endoscopic revision. The success rate of DCR for acquired NLDO in our group of patients was 77%, lower than reported in previous studies, with endoscopic surgery showing better results. Success rates of revision surgery were relatively low (<50%), and patients who fail the first revision are not likely to benefit from additional revisions.


Effects of regular walking on cardiovascular risk factors and body composition in normoglycemic women and women with type 2 diabetes.
To examine the impact of a 12-week walking program on body composition and risk factors for cardiovascular disease in women with type 2 diabetes and in normoglycemic women with first-degree diabetic relatives. There were 11 postmenopausal women with type 2 diabetes and 20 normoglycemic women of similar age and BMI who were asked to walk 1 h per day on 5 days each week for 12 weeks. Fitness (estimated VO2max) was assessed with a 1.6-km walking test; body composition was measured by dual-energy X-ray absorptiometry; and sex hormone, metabolic, and lipid concentrations were measured in serum. After 12 weeks, estimated VO2max improved in both groups (P < 0.005). In the diabetic women, BMI and fat content of the upper body and android waist region decreased (P < 0.05). Concentrations of fasting blood glucose (P < 0.05) HbAlc (P < 0.05), total cholesterol (P < 0.005), and LDL cholesterol (P < 0.05) decreased, while HDL cholesterol and sex hormones were unchanged. In contrast, normoglycemic women failed to lose body fat after 12 weeks of exercise in a walking program. However, their HbAlc, total cholesterol, LDL cholesterol, sex hormone-binding globulin, and total testosterone concentrations decreased (P < 0.05). On pooling the data and including diabetes as a categorical grouping variable, stepwise multiple regression analysis indicated that the change in centralized body fat, but not the change in VO2max, was related to change in fasting blood glucose. Twelve weeks of walking increased the fitness of diabetic and normoglycemic women. Improvement of fasting blood glucose was related to the loss of centralized body fat rather than to improved fitness.


Glycine supplementation extends lifespan of male and female mice.
Diets low in methionine extend lifespan of rodents, though through unknown mechanisms. Glycine can mitigate methionine toxicity, and a small prior study has suggested that supplemental glycine could extend lifespan of Fischer 344 rats. We therefore evaluated the effects of an 8% glycine diet on lifespan and pathology of genetically heterogeneous mice in the context of the Interventions Testing Program. Elevated glycine led to a small (4%-6%) but statistically significant lifespan increase, as well as an increase in maximum lifespan, in both males (p = 0.002) and females (p < 0.001). Pooling across sex, glycine increased lifespan at each of the three independent sites, with significance at p = 0.01, 0.053, and 0.03, respectively. Glycine-supplemented females were lighter than controls, but there was no effect on weight in males. End-of-life necropsies suggested that glycine-treated mice were less likely than controls to die of pulmonary adenocarcinoma (p = 0.03). Of the 40 varieties of incidental pathology evaluated in these mice, none were increased to a significant degree by the glycine-supplemented diet. In parallel analyses of the same cohort, we found no benefits from TM5441 (an inhibitor of PAI-1, the primary inhibitor of tissue and urokinase plasminogen activators), inulin (a source of soluble fiber), or aspirin at either of two doses. Our glycine results strengthen the idea that modulation of dietary amino acid levels can increase healthy lifespan in mice, and provide a foundation for further investigation of dietary effects on aging and late-life diseases.


Protein-labeling effects in confocal laser scanning microscopy.
Confocal laser scanning microscopy (CLSM) is being increasingly used for observing protein uptake in porous chromatography resins. Recent CLSM studies have revealed the possible existence of a nondiffusive protein transport mechanism. Observing protein uptake with CLSM requires labeling the protein with a fluorescent probe. This study examines the effect of the probe identity on the subsequent CLSM adsorption profiles. The adsorption of lysozyme conjugated with different fluorescent probes (Cy5, BODIPY FL, Atto 635, and Atto 520) on SP Sepharose Fast Flow was measured using CLSM and zonal chromatography experiments. Results from zonal chromatography show that the retention time of lysozyme-dye conjugates differ significantly from unlabeled lysozyme. The change in retention of lysozyme upon conjugation with a fluorescent probe is consistent with the difference in net charge between the lysozyme-dye conjugate and unlabeled lysozyme. The adsorption profiles measured by CLSM show significantly different behavior depending upon whether the lysozyme-dye conjugate is retained longer or shorter than the unlabeled lysozyme. These results strongly suggest that the lysozyme concentration overshoot observed in previous CLSM experiments is the result of displacement of weaker binding labeled lysozyme by stronger binding unlabeled lysozyme.


Integrating a sense of coherence into the neonatal environment.
Family centred care (FCC) is currently a valued philosophy within neonatal care; an approach that places the parents at the heart of all decision-making and engagement in the care of their infant. However, to date, there is a lack of clarity regarding the definition of FCC and limited evidence of FCCs effectiveness in relation to parental, infant or staff outcomes. In this paper we present a new perspective to neonatal care based on Aaron Antonovksy's Sense of Coherence (SOC) theory of well-being and positive health. Whilst the SOC was originally conceptualised as a psychological-based construct, the SOCs three underpinning concepts of comprehensibility, manageability and meaningfulness provide a theoretical lens through which to consider and reflect upon meaningful care provision in this particular care environment. By drawing on available FCC research, we consider how the SOC concepts considered from both a parental and professional perspective need to be addressed. The debate offered in this paper is not presented to reduce the importance or significance of FCC within neonatal care, but, rather, how consideration of the SOC offers the basis through which meaningful and effective FCC may be delivered. Practice based implications contextualised within the SOC constructs are also detailed. Consideration of the SOC constructs from both a parental and professional perspective need to be addressed in FCC provision. Service delivery and care practices need to be comprehensible, meaningful and manageable in order to achieve and promote positive well-being and health for all concerned.


[Congenital Heart Disease in Children with Down Syndrome: What Has Changed in the Last Three Decades?]
The prevalence of Down syndrome has increased in the last 30 years; 55% of these children have congenital heart disease. A retrospective longitudinal cohort study; clinical data from 1982 to 2013 databases with the diagnosis of Down syndrome or trisomy 21 in a reference hospital in pediatric cardiology and cardiac surgery. to assess the progress in the last three decades of cardiological care given to children with Down syndrome and congenital heart disease. We studied 102 patients with Down syndrome and congenital heart disease subjected to invasive therapy: corrective or palliative cardiac surgery and therapeutic catheterization. The referral age was progressively earlier in patients referred in the first year of life. The most frequent diagnosis was complete atrioventricular sptal defect (41%). There was a trend towards increasingly early corrective surgery in patients under 12 months (p < 0.001). Since 2000, the large majority of patients were operated before reaching six months of age. The main cardiac complications were rhythm dysfunction and low output. More frequent noncardiac complications were pulmonary and infectious. The 30-day mortality rate was 3/102 cases (2.9%). Of patients in follow-up, 89% are in NYHA class I. The early surgical correction seen over the past 15 years follows the approach suggested in the literature. The observed 30-day mortality rate is overlapping international results. Patients with Down syndrome subjected to corrective surgery of congenital heart disease have an excellent long-term functional capacity.


Rmi1 stimulates decatenation of double Holliday junctions during dissolution by Sgs1-Top3.
A double Holliday junction (dHJ) is a central intermediate of homologous recombination that can be processed to yield crossover or non-crossover recombination products. To preserve genomic integrity, cells possess mechanisms to avoid crossing over. We show that Saccharomyces cerevisiae Sgs1 and Top3 proteins are sufficient to migrate and disentangle a dHJ to produce exclusively non-crossover recombination products, in a reaction termed "dissolution." We show that Rmi1 stimulates dHJ dissolution at low Sgs1-Top3 protein concentrations, although it has no effect on the initial rate of Holliday junction (HJ) migration. Rmi1 serves to stimulate DNA decatenation, removing the last linkages between the repaired and template DNA molecules. Dissolution of a dHJ is a highly efficient and concerted alternative to nucleolytic resolution that prevents crossing over of chromosomes during recombinational DNA repair in mitotic cells and thereby contributes to genomic integrity.


A Context-Aware Application to Increase Elderly Users Compliance with Physical Rehabilitation Exercises at Home via Animatronic Biofeedback.
Biofeedback from physical rehabilitation exercises has proved to lead to faster recovery, better outcomes, and increased patient motivation. In addition, it allows the physical rehabilitation processes carried out at the clinic to be complemented with exercises performed at home. However, currently existing approaches rely mostly on audio and visual reinforcement cues, usually presented to the user on a computer screen or a mobile phone interface. Some users, such as elderly people, can experience difficulties to use and understand these interfaces, leading to non-compliance with the rehabilitation exercises. To overcome this barrier, latest biosignal technologies can be used to enhance the efficacy of the biofeedback, decreasing the complexity of the user interface. In this paper we propose and validate a context-aware framework for the use of animatronic biofeedback, as a way of potentially increasing the compliance of elderly users with physical rehabilitation exercises performed at home. In the scope of our work, animatronic biofeedback entails the use of pre-programmed actions on a robot that are triggered in response to certain changes detected in the users biomechanical or electrophysiological signals. We use electromyographic and accelerometer signals, collected in real time, to monitor the performance of the user while executing the exercises, and a mobile robot to provide animatronic reinforcement cues associated with their correct or incorrect execution. A context-aware application running on a smartphone aggregates the sensor data and controls the animatronic feedback. The acceptability of the animatronic biofeedback has been tested on a set of volunteer elderly users, and results suggest that the participants found the animatronic feedback engaging and of added value.


A comparative molecular and 3-dimensional structural investigation into cross-continental and novel avian Trypanosoma spp. in Australia.
Molecular and structural information on avian Trypanosoma spp. throughout Australia is limited despite their intrinsic value in understanding trypanosomatid evolution, diversity, and structural biology. In Western Australia tissue samples (n = 429) extracted from 93 birds in 25 bird species were screened using generic PCR primers to investigate the diversity of Trypanosoma spp. To investigate avian trypanosome structural biology the first 3-dimensional ultrastructural models of a Trypanosoma spp. (Trypanosoma sp. AAT) isolated from a bird (currawong, Strepera spp.) were generated using focussed ion beam milling combined with scanning electron microscopy (FIB-SEM). Here, we confirm four intercontinental species of avian trypanosomes in native Australian birds, and identify a new avian Trypanosoma. Trypanosome infection was identified in 18 birds from 13 different bird species (19%). A single new genotype was isolated and found to be closely related to T. culicavium (Trypanosoma sp. CC2016 B002). Other Trypanosoma spp. identified include T. avium, T. culicavium, T. thomasbancrofti, Trypanosoma sp. TL.AQ.22, Trypanosoma sp. AAT, and an uncharacterised Trypanosoma sp. (group C-III sensu Zidková et al. (Infect Genet Evol 12:102-112, 2012)), all previously identified in Australia or other continents. Serially-sectioning Trypanosoma sp. AAT epimastigotes using FIB-SEM revealed the disc-shaped kinetoplast pocket attached perpendicular to the branching mitochondrion. Additionally, the universal minicircle sequence within the kinetoplast DNA and the associated binding protein were determined in Trypanosoma sp. AAT. These results indicate that bird trypanosomes are relatively conserved across continents, while being locally diverse, which supports the hypothesis that bird trypanosomes exist as fewer species than described in the literature. Evidence exists that avian Trypanosoma spp. are infecting mammals and could be transmitted by haemadipsid leeches. Trypanosoma sp. AAT is most likely a separate species currently found only in Australia and the first 3-dimentional ultrastructural analysis of an avian trypanosome provides interesting information on their morphology and organelle arrangement.


The modified 2VO ischemia protocol causes cognitive impairment similar to that induced by the standard method, but with a better survival rate.
Permanent bilateral occlusion of the common carotid arteries (2VO) in the rat has been established as a valid experimental model to investigate the effects of chronic cerebral hypoperfusion on cognitive function and neurodegenerative processes. Our aim was to compare the cognitive and morphological outcomes following the standard 2VO procedure, in which there is concomitant artery ligation, with those of a modified protocol, with a 1-week interval between artery occlusions to avoid an abrupt reduction of cerebral blood flow, as assessed by animal performance in the water maze and damage extension to the hippocampus and striatum. Male Wistar rats (N = 47) aged 3 months were subjected to chronic hypoperfusion by permanent bilateral ligation of the common carotid arteries using either the standard or the modified protocol, with the right carotid being the first to be occluded. Three months after the surgical procedure, rat performance in the water maze was assessed to investigate long-term effects on spatial learning and memory and their brains were processed in order to estimate hippocampal volume and striatal area. Both groups of hypoperfused rats showed deficits in reference (F(₈,₁₇₂) = 7.0951, P < 0.00001) and working spatial memory [2nd (F(₂,₄₄) = 7.6884, P < 0.001), 3rd (F(₂,₄₄) = 21.481, P < 0.00001) and 4th trials (F(₂,₄₄) = 28.620, P < 0.0001)]; however, no evidence of tissue atrophy was found in the brain structures studied. Despite similar behavioral and morphological outcomes, the rats submitted to the modified protocol showed a significant increase in survival rate, during the 3 months of the experiment (P < 0.02).


Mental health symptoms during imprisonment: a longitudinal study.
Mental health problems are common in prison populations. Less is known about how mental health problems develop during imprisonment. The objective was to examine the longitudinal course of mental health symptoms during imprisonment and individual factors associated with the development of these symptoms. In a prospective cohort study, 1.664 Dutch male prisoners were questioned 3 weeks after their arrival in detention. Those still in custody were questioned again after 3, 9, and 18 months. Multilevel analyses were conducted to identify predictors of the course of mental health symptoms. Prisoners reported continued elevated symptom levels compared to the general population. Inmates who entered detention with pre-existing mental health problems and problematic alcohol/drugs use showed mental health improvements during detention. The high levels of prisoners' mental health problems highlight the importance of addressing mental health issues in prison. Imprisonment does not have an overall negative effect on mental health.


Quantum dot-based sensitive detection of disease specific exosome in serum.
Tumor-derived exosomes have emerged as promising cancer biomarkers due to their unique composition and functions. Herein, we report a stripping voltammetric immunoassay for the electrochemical detection of disease-specific exosomes using quantum dots as signal amplifiers. The assay involves three subsequent steps where bulk exosome populations are initially magnetically captured on magnetic beads by a generic tetraspanin antibody (e.g., CD9 or CD63) followed by the identification of disease-specific exosomes using cancer-related. Here, we used CdSe quantum dot (CdSeQD) functionalised-biotinylated HER-2 and FAM134B antibodies as breast and colon cancer markers. After magnetic washing and purification steps, acid dissolution of CdSeQDs and subsequent anodic stripping voltammetric quantification of Cd2+ were carried out at the bare glassy carbon working electrode. This method enabled sensitive detection of 100 exosomes per μL with a relative standard deviation (%RSD) of <5.5% in cancer cell lines and a small cohort of serum samples (n = 9) collected from patients with colorectal adenocarcinoma. We believe that our approach could potentially represent an effective bioassay for the quantification of disease-specific exosomes in clinical samples.


Thrombelastography change after bridging to left ventricular assist device from extracorporeal membrane oxygenation patients.
Circulatory assist devices are applied to support patients with end-stage heart failure or circulatory collapse. Extracorporeal membrane oxygenation (ECMO) and ventricular assist device (VAD) are the two devices that are commonly used in these situations. However, in patients undergoing ECMO, complications occur more frequently and are more severe than in those with VAD, especially in patients with coagulopathy. The cause of coagulopathy is seldom reported; therefore, the current prospective study was designed to compare the coagulation status between patients who received ECMO and bridged VAD thereafter. The cases for our study were collected between December 2005 and January 2010. A total of 21 patients with VAD were enrolled. Seven patients received initial ECMO support and were subsequently shifted to VAD. Use of blood transfusion products by patients was recorded during application of ECMO and VAD. Thrombelastography (TEG) was performed pre-ECMO and pre-VAD, and thereafter to 1 week post-VAD implantation. The relevant parameters that were examined included coagulation time (R: resting, K: kinetics, seconds), angle (α, degrees), maximal amplitude (MA, mm), clot strength (G, dynes/cm(2)), and estimated percent lysis in 30 minutes (LY30, %). Coagulation status and average unit per day of blood transfusion at pre-VAD and post-VAD status were recorded. TEGs of pre-VAD and post-VAD implantation were compared in order to determine the frequency of bleeding. Average results from seven cases were incorporated into the current analysis. The patient transfusion requirement was reduced after implantation of the VAD. The TEG of post-VAD implantation was improved from the earlier pre-VAD results (ECMO status). Ultimately, TEG resulted in significant differences in R, K, angle, MA, and G, postoperatively. Coagulopathy of patients with VAD improved after bridging from ECMO, which can be attributed to improved coagulation factor and platelet function. Thus, bleeding complications are reduced.


Evaluation of the Azure Kinect and Its Comparison to Kinect V1 and Kinect V2.
The Azure Kinect is the successor of Kinect v1 and Kinect v2. In this paper we perform brief data analysis and comparison of all Kinect versions with focus on precision (repeatability) and various aspects of noise of these three sensors. Then we thoroughly evaluate the new Azure Kinect; namely its warm-up time, precision (and sources of its variability), accuracy (thoroughly, using a robotic arm), reflectivity (using 18 different materials), and the multipath and flying pixel phenomenon. Furthermore, we validate its performance in both indoor and outdoor environments, including direct and indirect sun conditions. We conclude with a discussion on its improvements in the context of the evolution of the Kinect sensor. It was shown that it is crucial to choose well designed experiments to measure accuracy, since the RGB and depth camera are not aligned. Our measurements confirm the officially stated values, namely standard deviation ≤17 mm, and distance error <11 mm in up to 3.5 meters distance from the sensor in all four supported modes. The device, however, has to be warmed up for at least 40-50 min to give stable results. Due to the time-of-flight technology, the Azure Kinect cannot be reliably used in direct sunlight. Therefore, it is convenient mostly for indoor applications.


Efficient delivery of small interfering RNA to bone-metastatic tumors by using atelocollagen in vivo.
Silencing of gene expression by small interfering RNAs (siRNAs) is rapidly becoming a powerful tool for genetic analysis and represents a potential strategy for therapeutic product development. However, there are no reports of systemic delivery for siRNAs toward treatment of bone-metastatic cancer. Accordingly, we report here that i.v. injection of GL3 luciferase siRNA complexed with atelocollagen showed effective reduction of luciferase expression from bone-metastatic prostate tumor cells developed in mouse thorax, jaws, and/or legs. We also show that the siRNA/atelocollagen complex can be efficiently delivered to tumors 24 h after injection and can exist intact at least for 3 days. Furthermore, atelocollagen-mediated systemic administration of siRNAs such as enhancer of zeste homolog 2 and phosphoinositide 3'-hydroxykinase p110-alpha-subunit, which were selected as candidate targets for inhibition of bone metastasis, resulted in an efficient inhibition of metastatic tumor growth in bone tissues. In addition, upregulation of serum IL-12 and IFN-alpha levels was not associated with the in vivo administration of the siRNA/atelocollagen complex. Thus, for treatment of bone metastasis of prostate cancer, an atelocollagen-mediated systemic delivery method could be a reliable and safe approach to the achievement of maximal function of siRNA.


Communication: the influence of CO2 poisoning on overvoltages and discharge capacity in non-aqueous Li-Air batteries.
The effects of Li2CO3 like species originating from reactions between CO2 and Li2O2 at the cathode of non-aqueous Li-air batteries were studied by density functional theory (DFT) and galvanostatic charge-discharge measurements. Adsorption energies of CO2 at various nucleation sites on a stepped (11̅00) Li2O2 surface were determined and even a low concentration of CO2 effectively blocks the step nucleation site and alters the Li2O2 shape due to Li2CO3 formation. Nudged elastic band calculations show that once CO2 is adsorbed on a step valley site, it is effectively unable to diffuse and impacts the Li2O2 growth mechanism, capacity, and overvoltages. The charging processes are strongly influenced by CO2 contamination, and exhibit increased overvoltages and increased capacity, as a result of poisoning of nucleation sites: this effect is predicted from DFT calculations and observed experimentally already at 1% CO2. Large capacity losses and overvoltages are seen at higher CO2 concentrations.


Molecularly imprinted polymers for the analysis and removal of polychlorinated aromatic compounds in the environment: a review.
Synthetic receptors and in particular molecularly imprinted polymers (MIPs) are gaining relevance as selective sorbent materials and biomimetic recognition elements for analyzing polychlorinated aromatic compounds (PACs) in the environment. PACs are still ubiquitous toxic pollutants requiring their continuous environmental assessment for protecting humans and animals from exposure. Since nowadays most PACs occur at ultra-trace concentration levels and in complex matrices, the selectivity of MIPs renders them ideally suited for facilitating either sample pre-treatment and quantitative enrichment, or acting as biomimetic recognition elements as an integral component of corresponding sensing schemes. Due to the diversity of PACs, imprinting polymers for these constituents appears particularly challenging. This review focuses on prevalent strategies towards successfully templating polymer materials towards polychlorinated biphenyls and their hydroxy forms, chlorophenols, dioxins and furans, and organochlorine pesticides, and successful applications of the polymer materials in monitoring of these compounds at trace-levels in real-world environmental matrices. Discussed are also group-selective sorbents for facilitating simultaneous detection and quantification of PACs.


Morphiceptin (NH4-tyr-pro-phe-pro-COHN2): a potent and specific agonist for morphine (mu) receptors.
The synthetic peptide NH2-Tyr-Pro-Phe-Pro-CONH2 (morphiceptin), which is the amide of a fragment of the milk protein beta-casein, has morphinelike activities and is highly specific for morphine (mu) receptors but not for enkephalin (delta) receptors. It is as active as morphine in the guinea pig ileum but much less active in the mouse and rat vas deferens. The discovery of this specific morphine receptor ligand substantiates the hypothesis of multiple opiate receptors. The ligand, which may be of physiological significance since a very similar, or identical, activity can be detected in enzymatic digests of beta-casein, may prove useful for further investigation of the functions of opiate receptor subtypes.


Prion replication without host adaptation during interspecies transmissions.
Adaptation of prions to new species is thought to reflect the capacity of the host-encoded cellular form of the prion protein (PrPC) to selectively propagate optimized prion conformations from larger ensembles generated in the species of origin. Here we describe an alternate replicative process, termed nonadaptive prion amplification (NAPA), in which dominant conformers bypass this requirement during particular interspecies transmissions. To model susceptibility of horses to prions, we produced transgenic (Tg) mice expressing cognate PrPC Although disease transmission to only a subset of infected TgEq indicated a significant barrier to EqPrPC conversion, the resulting horse prions unexpectedly failed to cause disease upon further passage to TgEq. TgD expressing deer PrPC was similarly refractory to deer prions from diseased TgD infected with mink prions. In both cases, the resulting prions transmitted to mice expressing PrPC from the species of prion origin, demonstrating that transmission barrier eradication of the originating prions was ephemeral and adaptation superficial in TgEq and TgD. Horse prions produced in vitro by protein misfolding cyclic amplification of mouse prions using horse PrPC also failed to infect TgEq but retained tropism for wild-type mice. Concordant patterns of neuropathology and prion deposition in susceptible mice infected with NAPA prions and the corresponding prion of origin confirmed preservation of strain properties. The comparable responses of both prion types to guanidine hydrochloride denaturation indicated this occurs because NAPA precludes selection of novel prion conformations. Our findings provide insights into mechanisms regulating interspecies prion transmission and a framework to reconcile puzzling epidemiological features of certain prion disorders.


Adolescent sexuality and positive well-being: a group-norms approach.
The link between adolescent sexual activity and psychological well-being is a controversial issue in developmental psychology. This cross-sectional study investigated the association between three aspects of teenage sexuality (genital sexual experience, age of sexual onset, and number of sex partners) and positive well-being (hedonic, eudaimonic, and overall) in a sample of 475 high school seniors (48% female; 89% White) from a single school district in a rural upstate New York community. Based on a group-norms perspective, we expected higher well-being in adolescents whose sexual behaviors followed group-normative patterns. As expected, sexually experienced and on-time (at age 16) students reported higher well-being than sexually inexperienced or late-onset (17 or older) students. Contrary to expectations, a high number of sex partners and an early sexual onset (15 or younger) were not related to lower well-being. Early-onset girls reported higher levels of well-being than normative-onset peers. Findings are discussed in relationship to theoretical perspectives and past empirical findings of teenage sexuality as a developmental asset versus risk.


Nitrate enhances skeletal muscle fatty acid oxidation via a nitric oxide-cGMP-PPAR-mediated mechanism.
Insulin sensitivity in skeletal muscle is associated with metabolic flexibility, including a high capacity to increase fatty acid (FA) oxidation in response to increased lipid supply. Lipid overload, however, can result in incomplete FA oxidation and accumulation of potentially harmful intermediates where mitochondrial tricarboxylic acid cycle capacity cannot keep pace with rates of β-oxidation. Enhancement of muscle FA oxidation in combination with mitochondrial biogenesis is therefore emerging as a strategy to treat metabolic disease. Dietary inorganic nitrate was recently shown to reverse aspects of the metabolic syndrome in rodents by as yet incompletely defined mechanisms. Herein, we report that nitrate enhances skeletal muscle FA oxidation in rodents in a dose-dependent manner. We show that nitrate induces FA oxidation through a soluble guanylate cyclase (sGC)/cGMP-mediated PPARβ/δ- and PPARα-dependent mechanism. Enhanced PPARβ/δ and PPARα expression and DNA binding induces expression of FA oxidation enzymes, increasing muscle carnitine and lowering tissue malonyl-CoA concentrations, thereby supporting intra-mitochondrial pathways of FA oxidation and enhancing mitochondrial respiration. At higher doses, nitrate induces mitochondrial biogenesis, further increasing FA oxidation and lowering long-chain FA concentrations. Meanwhile, nitrate did not affect mitochondrial FA oxidation in PPARα(-/-) mice. In C2C12 myotubes, nitrate increased expression of the PPARα targets Cpt1b, Acadl, Hadh and Ucp3, and enhanced oxidative phosphorylation rates with palmitoyl-carnitine; however, these changes in gene expression and respiration were prevented by inhibition of either sGC or protein kinase G. Elevation of cGMP, via the inhibition of phosphodiesterase 5 by sildenafil, also increased expression of Cpt1b, Acadl and Ucp3, as well as CPT1B protein levels, and further enhanced the effect of nitrate supplementation. Nitrate may therefore be effective in the treatment of metabolic disease by inducing FA oxidation in muscle.


Dietary supplements for aquatic sports.
Many athletes use dietary supplements, with use more prevalent among those competing at the highest level. Supplements are often self-prescribed, and their use is likely to be based on an inadequate understanding of the issues at stake. Supplementation with essential micronutrients may be useful when a diagnosed deficiency cannot be promptly and effectively corrected with food-based dietary solutions. When used in high doses, some supplements may do more harm than good: Iron supplementation, for example, is potentially harmful. There is good evidence from laboratory studies and some evidence from field studies to support health or performance benefits from appropriate use of a few supplements. The available evidence from studies of aquatic sports is small and is often contradictory. Evidence from elite performers is almost entirely absent, but some athletes may benefit from informed use of creatine, caffeine, and buffering agents. Poor quality assurance in some parts of the dietary supplements industry raises concerns about the safety of some products. Some do not contain the active ingredients listed on the label, and some contain toxic substances, including prescription drugs, that can cause health problems. Some supplements contain compounds that will cause an athlete to fail a doping test. Supplement quality assurance programs can reduce, but not entirely eliminate, this risk.


An approach to characterizing single-subunit mutations in multimeric prepores and pores of anthrax protective antigen.
Heptameric pores formed by the protective antigen (PA) moiety of anthrax toxin translocate the intracellular effector moieties of the toxin across the endosomal membrane to the cytosol of mammalian cells. We devised a protocol to characterize the effects of individual mutations in a single subunit of heptameric PA prepores (pore precursors) or pores. We prepared monomeric PA containing a test mutation plus an innocuous Cys-replacement mutation at a second residue (Lys563, located on the external surface of the prepore). The introduced Cys was biotinylated, and the protein was allowed to cooligomerize with a 20-fold excess of wild-type PA. Finally, biotinylated prepores were freed from wild-type prepores by avidin affinity chromatography. For the proof of principle, we examined single-subunit mutations of Asp425 and Phe427, two residues where Ala replacements have been shown to cause strong inhibitory effects. The single-subunit D425A mutation inhibited pore formation by >10(4) and abrogated activity of PA almost completely in our standard cytotoxicity assay. The single-subunit F427A mutation caused approximately 100-fold inhibition in the cytotoxicity assay, and this effect was shown to result from a combination of strong inhibition of translocation and smaller effects on pore formation and ligand affinity. Our results show definitively that replacing a single residue in one subunit of the heptameric PA prepore can inhibit the transport activity of the oligomer almost completely-and by different mechanisms, depending on the specific residue mutated.


Use of local intrinsic muscle flaps for diabetic foot and ankle reconstruction: a systematic review.
To assess the outcomes of local intrinsic muscle flaps specifically in the surgical reconstruction of foot and ankle wounds in patients with diabetes. A systematic review of the literature was conducted by three independent reviewers using several databases and search engines. The following search terms were used: foot, ankle, diabetes mellitus, peripheral neuropathy, Charcot neuroarthropathy, ulceration, wound, muscle, flap, surgical offloading, diabetic reconstruction, local intrinsic, abductor digiti minimi, abductor hallucis, extensor digitorum brevis, flexor digitorum brevis, flexor digiti minimi. Studies meeting the following criteria were included: English language studies, studies published from 1997 to 2017 (inclusive of the last 20 years), patients with diabetes treated for foot and/or ankle wounds, use of local intrinsic muscle flaps, follow-up period of six months or more postoperatively, documentation of healing rates, complications, and need for revisional surgery. Studies that were entirely literature reviews, descriptions of only surgical technique and/or cadaveric studies and patients without diabetes were excluded. A total of 13 selected studies met the above inclusion criteria and were used for data extraction from a total of 113 patients in which 92 local intrinsic muscle flaps were performed for diabetic foot and/or ankle reconstruction, of which 80 (87%) muscle flaps demonstrated success, while 12 (13%) flaps encountered complications. Although local intrinsic muscle flaps demonstrated a relatively high success rate when used for the definitive closure of certain diabetic foot and ankle wounds based on existing evidence, our systematic review emphasises the need for more high-quality, larger comparative studies to investigate their efficacy and failure rates in diabetic foot and/or ankle reconstruction.


A study of optical design and optimization of zoom optics with liquid lenses through modified genetic algorithm.
A new concept for the optimization and optical design of miniature digital zoom optics with liquid lens elements is proposed in this research. The liquid lens elements are limited to the discrete configuration in order to obtain the optimal performance for digital zoom. We propose a newly developed digital zoom layout and optimization with a modified genetic algorism (GA) method, in order to meet the demands of a certain specification. The results show that we achieve a successful optical design and the optimization of the digital zoom optics with liquid optics, whose performance is greatly improved up to 48.68%, from the standpoint of on-axis spot size.


DNA damage checkpoint triggers autophagy to regulate the initiation of anaphase.
Budding yeast cells suffering a single unrepaired double-strand break (DSB) trigger the Mec1 (ATR)-dependent DNA damage response that causes them to arrest before anaphase for 12-15 h. Here we find that hyperactivation of the cytoplasm-to-vacuole (CVT) autophagy pathway causes the permanent G2/M arrest of cells with a single DSB that is reflected in the nuclear exclusion of both Esp1 and Pds1. Transient relocalization of Pds1 is also seen in wild-type cells lacking vacuolar protease activity after induction of a DSB. Arrest persists even as the DNA damage-dependent phosphorylation of Rad53 diminishes. Permanent arrest can be overcome by blocking autophagy, by deleting the vacuolar protease Prb1, or by driving Esp1 into the nucleus with a SV40 nuclear localization signal. Autophagy in response to DNA damage can be induced in three different ways: by deleting the Golgi-associated retrograde protein complex (GARP), by adding rapamycin, or by overexpression of a dominant ATG13-8SA mutation.


Multi-university research teams: shifting impact, geography, and stratification in science.
This paper demonstrates that teamwork in science increasingly spans university boundaries, a dramatic shift in knowledge production that generalizes across virtually all fields of science, engineering, and social science. Moreover, elite universities play a dominant role in this shift. By examining 4.2 million papers published over three decades, we found that multi-university collaborations (i) are the fastest growing type of authorship structure, (ii) produce the highest-impact papers when they include a top-tier university, and (iii) are increasingly stratified by in-group university rank. Despite the rising frequency of research that crosses university boundaries, the intensification of social stratification in multi-university collaborations suggests a concentration of the production of scientific knowledge in fewer rather than more centers of high-impact science.


Characteristics and consequences of missed gastric cancer: A multicentric cohort study.
Missed gastric cancer (MGC) is poorly documented in Mediterranean populations. (1) To assess the rate, predictors and survival of MGC. (2) To compare MGC and non-MGC tumors. This is a retrospective-cohort study conducted at four centers. MGC was defined as cancer detected within three years after negative esophagogastroduodenoscopy. Gastric adenocarcinomas diagnosed between 2008-2015 were included. Patients with no follow-up were excluded. During the study period 123,395 esophagogastroduodenoscopies were performed, with 1374 gastric cancers being diagnosed (1.1%). A total of 1289 gastric cancers were finally included. The overall rate of MGC was 4.7% (61/1289, 3.7-6%). A negative esophagogastroduodenoscopy in MGC patients was independently associated with PPI therapy (p < 0.001), previous Billroth II anastomosis (p = 0.002), and lack of alarm symptoms (p < 0.001). The most frequent location for MGC was the gastric body(52.4%). MGCs were smaller than non-MGCs (31 vs 41 mm, p = 0.047), more often flat or depressed (p = 0.003) and less likely to be encountered as advanced disease. Overall 2-year survival was similar between MGC (34.1%) and Non-MGC (35.3 %) (p = 0.59). MGC accounted for nearly five percent of newly-diagnosed gastric adenocarcinomas. Overall survival was poor and not different between MGC and non-MGC.


Oxidant stress evoked by pacemaking in dopaminergic neurons is attenuated by DJ-1.
Parkinson's disease is a pervasive, ageing-related neurodegenerative disease the cardinal motor symptoms of which reflect the loss of a small group of neurons, the dopaminergic neurons in the substantia nigra pars compacta (SNc). Mitochondrial oxidant stress is widely viewed as being responsible for this loss, but why these particular neurons should be stressed is a mystery. Here we show, using transgenic mice that expressed a redox-sensitive variant of green fluorescent protein targeted to the mitochondrial matrix, that the engagement of plasma membrane L-type calcium channels during normal autonomous pacemaking created an oxidant stress that was specific to vulnerable SNc dopaminergic neurons. The oxidant stress engaged defences that induced transient, mild mitochondrial depolarization or uncoupling. The mild uncoupling was not affected by deletion of cyclophilin D, which is a component of the permeability transition pore, but was attenuated by genipin and purine nucleotides, which are antagonists of cloned uncoupling proteins. Knocking out DJ-1 (also known as PARK7 in humans and Park7 in mice), which is a gene associated with an early-onset form of Parkinson's disease, downregulated the expression of two uncoupling proteins (UCP4 (SLC25A27) and UCP5 (SLC25A14)), compromised calcium-induced uncoupling and increased oxidation of matrix proteins specifically in SNc dopaminergic neurons. Because drugs approved for human use can antagonize calcium entry through L-type channels, these results point to a novel neuroprotective strategy for both idiopathic and familial forms of Parkinson's disease.


The concept of patient satisfaction in adolescent psychiatric care: a qualitative study.
Few studies have asked how adolescents perceive the quality of psychiatric care. Therefore, the aim of the study was to explore adolescents' perception of quality of care and of satisfying treatment outcomes. Fourteen adolescents participated in this hermeneutic study. Several concepts describing adolescents' perspective of quality of care were found: secure place, tough love, peer solidarity, self-expression, and person not patient. Concepts describing satisfying treatment outcomes fell into four categories: improved mental health, personal development, strengthening of the self, and improved family relations. By casting light on users' perspectives, the study offers guidance for improvement of quality of care and for the development of patient satisfaction instruments.


The impact of neuroscience on society: cognitive enhancement in neuropsychiatric disorders and in healthy people.
In addition to causing distress and disability to the individual, neuropsychiatric disorders are also extremely expensive to society and governments. These disorders are both common and debilitating and impact on cognition, functionality and wellbeing. Cognitive enhancing drugs, such as cholinesterase inhibitors and methylphenidate, are used to treat cognitive dysfunction in Alzheimer's disease and attention deficit hyperactivity disorder, respectively. Other cognitive enhancers include specific computerized cognitive training and devices. An example of a novel form of cognitive enhancement using the technological advancement of a game on an iPad that also acts to increase motivation is presented. Cognitive enhancing drugs, such as methylphenidate and modafinil, which were developed as treatments, are increasingly being used by healthy people. Modafinil not only affects 'cold' cognition, but also improves 'hot' cognition, such as emotion recognition and task-related motivation. The lifestyle use of 'smart drugs' raises both safety concerns as well as ethical issues, including coercion and increasing disparity in society. As a society, we need to consider which forms of cognitive enhancement (e.g. pharmacological, exercise, lifelong learning) are acceptable and for which groups (e.g. military, doctors) under what conditions (e.g. war, shift work) and by what methods we would wish to improve and flourish.


Classification of patient-safety incidents in primary care.
Primary care lags behind secondary care in the reporting of, and learning from, incidents that put patient safety at risk. In primary care, there is no universally agreed approach to classifying the severity of harm arising from such patient-safety incidents. This lack of an agreed approach limits learning that could lead to the prevention of injury to patients. In a review of research on patient safety in primary care, we identified 21 existing approaches to the classification of harm severity. Using the World Health Organization's (WHO's) International Classification for Patient Safety as a reference, we undertook a framework analysis of these approaches. We then developed a new system for the classification of harm severity. To assess and classify harm, most existing approaches use measures of symptom duration (11/21), symptom severity (11/21) and/or the level of intervention required to manage the harm (14/21). However, few of these approaches account for the deleterious effects of hospitalization or the psychological stress that may be experienced by patients and/or their relatives. The new classification system we developed builds on WHO's International Classification for Patient Safety and takes account not only of hospitalization and psychological stress but also of so-called near misses and uncertain outcomes. The constructs we have outlined have the potential to be applied internationally, across primary-care settings, to improve both the detection and prevention of incidents that cause the most severe harm to patients.


AGE challenge: Angle Closure Glaucoma Evaluation in Anterior Segment Optical Coherence Tomography.
Angle closure glaucoma (ACG) is a more aggressive disease than open-angle glaucoma, where the abnormal anatomical structures of the anterior chamber angle (ACA) may cause an elevated intraocular pressure and gradually lead to glaucomatous optic neuropathy and eventually to visual impairment and blindness. Anterior Segment Optical Coherence Tomography (AS-OCT) imaging provides a fast and contactless way to discriminate angle closure from open angle. Although many medical image analysis algorithms have been developed for glaucoma diagnosis, only a few studies have focused on AS-OCT imaging. In particular, there is no public AS-OCT dataset available for evaluating the existing methods in a uniform way, which limits progress in the development of automated techniques for angle closure detection and assessment. To address this, we organized the Angle closure Glaucoma Evaluation challenge (AGE), held in conjunction with MICCAI 2019. The AGE challenge consisted of two tasks: scleral spur localization and angle closure classification. For this challenge, we released a large dataset of 4800 annotated AS-OCT images from 199 patients, and also proposed an evaluation framework to benchmark and compare different models. During the AGE challenge, over 200 teams registered online, and more than 1100 results were submitted for online evaluation. Finally, eight teams participated in the onsite challenge. In this paper, we summarize these eight onsite challenge methods and analyze their corresponding results for the two tasks. We further discuss limitations and future directions. In the AGE challenge, the top-performing approach had an average Euclidean Distance of 10 pixels (10 µm) in scleral spur localization, while in the task of angle closure classification, all the algorithms achieved satisfactory performances, with two best obtaining an accuracy rate of 100%. These artificial intelligence techniques have the potential to promote new developments in AS-OCT image analysis and image-based angle closure glaucoma assessment in particular.


Entry and integration of transplanted hepatocytes in rat liver plates occur by disruption of hepatic sinusoidal endothelium.
To establish the process by which transplanted cells integrate into the liver parenchyma, we used dipeptidyl peptidase IV-deficient F344 rats as hosts. On intrasplenic injection, transplanted hepatocytes immediately entered liver sinusoids, along with attenuation of portal vein radicles on angiography. However, a large fraction of transplanted cells (>70%) was rapidly cleared from portal spaces by phagocyte/macrophage responses. On the other hand, transplanted hepatocytes entering the hepatic sinusoids showed superior survival. These cells translocated from sinusoids into liver plates between 16 and 20 hours after transplantation, during which electron microscopy showed disruption of the sinusoidal endothelium. Interestingly, production of vascular endothelial growth factor was observed in hepatocytes before endothelial disruptions. Portal hypertension and angiographic changes resulting from cell transplantation resolved promptly. Integration of transplanted hepatocytes in the liver parenchyma required cell membrane regenesis, with hybrid gap junctions and bile canaliculi forming over 3 to 7 days after cell transplantation. We propose that strategies to deposit cells into distal hepatic sinusoids, to disrupt sinusoidal endothelium for facilitating cell entry into liver plates, and to accelerate cell integrations into liver parenchyma will advance applications of hepatocyte transplantation.


Cerebellar plasticity and motor learning deficits in a copy-number variation mouse model of autism.
A common feature of autism spectrum disorder (ASD) is the impairment of motor control and learning, occurring in a majority of children with autism, consistent with perturbation in cerebellar function. Here we report alterations in motor behaviour and cerebellar synaptic plasticity in a mouse model (patDp/+) for the human 15q11-13 duplication, one of the most frequently observed genetic aberrations in autism. These mice show ASD-resembling social behaviour deficits. We find that in patDp/+ mice delay eyeblink conditioning--a form of cerebellum-dependent motor learning--is impaired, and observe deregulation of a putative cellular mechanism for motor learning, long-term depression (LTD) at parallel fibre-Purkinje cell synapses. Moreover, developmental elimination of surplus climbing fibres--a model for activity-dependent synaptic pruning--is impaired. These findings point to deficits in synaptic plasticity and pruning as potential causes for motor problems and abnormal circuit development in autism.


Episymbiotic microbes as food and defence for marine isopods: unique symbioses in a hostile environment.
Symbioses profoundly affect the diversity of life, often through novel biochemical services that symbionts provide to their hosts. These biochemical services are typically nutritional enhancements and less commonly defensive, but rarely both simultaneously. On the coral reefs of Papua New Guinea, we discovered unique associations between marine isopod crustaceans (Santia spp.) and episymbiotic microbes. Transmission electron microscopy and pigment analyses show that episymbiont biomass is dominated by large (20-30 microm) cyanobacterial cells. The isopods consume these photosymbionts and "cultivate" them by inhabiting exposed sunlit substrates, a behaviour made possible by symbionts' production of a chemical defence that is repulsive to fishes. Molecular phylogenetic analyses demonstrated that the symbiotic microbial communities are diverse and probably dominated in terms of population size by bacteria and small unicellular Synechococcus-type cyanobacteria. Although largely unknown in the oceans, defensive symbioses probably promote marine biodiversity by allowing niche expansions into otherwise hostile environments.


Pathologic gambling.
Pathologic gambling and problem gambling affect approximately 5 to 15 million Americans and are common in young people. The community-minded family physician is in a good position to identify and assist patients who have gambling-related problems and thereby prevent or treat the resultant personal, family and social disruptions. Provider and community education about the depth and breadth of this condition is crucial for the identification and treatment of a growing problem. As with many psychologic conditions, identification of the disorder and treatment of the patient by the family physician comprise the primary treatment. Screening tools, treatment programs and self-help groups provide additional resources for the family physician. An illustrative case report demonstrates the importance of heightened awareness of and screening for this common condition.


Evolution of the diverse biological roles of inositols.
Several of the nine hexahydroxycylohexanes (inositols) have functions in Biology, with myo-inositol (Ins) in most of the starring roles; and Ins polyphosphates are amongst the most abundant organic phosphate constituents on Earth. Many Archaea make Ins and use it as a component of diphytanyl membrane phospholipids and the thermoprotective solute di-L-Ins-1,1'-phosphate. Few bacteria make Ins or use it, other than as a carbon source. Those that do include hyperthermophilic Thermotogales (which also employ di-L-Ins-1,1'-phosphate) and actinomycetes such as Mycobacterium spp. (which use mycothiol, an inositol-containing thiol, as an intracellular redox reagent and have characteristic phosphatidylinositol-linked surface oligosaccharides). Bacteria acquired their Ins3P synthases by lateral gene transfer from Archaea. Many eukaryotes, including stressed plants, insects, deep-sea animals and kidney tubule cells, adapt to environmental variation by making or accumulating diverse inositol derivatives as 'compatible' solutes. Eukaryotes use phosphatidylinositol derivatives for numerous roles in cell signalling and regulation and in protein anchoring at the cell surface. Remarkably, the diradylglycerol cores of archaeal and eukaryote/bacterial glycerophospholipids have mirror image configurations: sn-2,3 and sn-1,2 respectively. Multicellular animals and amoebozoans exhibit the greatest variety of functions for PtdIns derivatives, including the use of PtdIns(3,4,5)P3 as a signal. Evolutionarily, it seems likely that (i) early archaeons first made myo-inositol approx. 3500 Ma (million years) ago; (ii) archeons brought inositol derivatives into early eukaryotes (approx. 2000 Ma?); (iii) soon thereafter, eukaryotes established ubiquitous functions for phosphoinositides in membrane trafficking and Ins polyphosphate synthesis; and (iv) since approx. 1000 Ma, further waves of functional diversification in amoebozoans and metazoans have introduced Ins(1,4,5)P3 receptor Ca2+ channels and the messenger role of PtdIns(3,4,5)P3.


Continuation of pregnancy after first-trimester exposure to mifepristone: an observational prospective study.
To report the follow-up of continuing pregnancies after first-trimester exposure to mifepristone. Observational prospective study. France. Patients exposed to mifepristone during the first 12 weeks of pregnancy. Women were included in the study when they or their doctors asked a French pharmacovigilance centre or the Paris Teratogen Information Service about the risk of mifepristone exposure in early pregnancy. Exclusion criteria were requests received after 22 weeks of gestation or subsequent elective termination of pregnancy without a pathological examination of the fetus. Data on maternal history and drug exposure were collected on first contact, and pregnancy outcomes were documented at follow-up. Rate of major congenital malformations. A total of 105 pregnancies were included, with 46 exposed to mifepristone alone, and 59 exposed to both mifepristone and misoprostol. There were 94 live births (90.4%) and 10 (9.6%) miscarriages (including one with major malformation). Elective termination of pregnancy was performed after the subsequent diagnosis of trisomy 21 in one case. The overall rate of major congenital malformations was 4.2% (95% CI 1.2-10.4%), with two cases among 38 patients exposed to mifepristone alone, and two cases among 57 patients exposed to both mifepristone and misoprostol. This first prospective study found that the rate of major malformations after first-trimester exposure to mifepristone is only slightly higher than the expected 2-3% rate in the general population. Such findings provide reassuring data for risk evaluation for continuation of pregnancy after mifepristone exposure.


Quality of inpatient care in public and private hospitals in Sri Lanka.
To compare the quality of inpatient clinical care in public and private hospitals in Sri Lanka. A retrospective, cross-sectional comparison was done of inpatient quality, in a sample of 11 public and 10 private hospitals in three of 25 districts. Data were collected for 55 quality indicators from medical records of 2523 public and 1815 private inpatient admissions. These covered treatment of asthma, acute myocardial infarction (AMI), childbirth and five other conditions, along with outcome indicators, and medicine prescribing indicators. Overall quality scores were better in the public sector than the private sector (77 vs 69%). Performance was similar for management of AMI and childbirth and somewhat better in the private sector for management of asthma. The public sector performed better in those indicators that are not constrained by resources (94 vs 81%), but worse in indicators that are highly resource intensive (10 vs 31%). Quality was comparable in assessment and investigation, but the public sector performed better in treatment and management (70 vs 62%) and drug prescribing (68 vs 60%), and modestly worse in terms of outcomes (92 vs 97%). For a range of indicators where comparisons were possible, quality of inpatient clinical care in Sri Lanka was comparable to levels reported from upper-middle income Asian countries, and often approaches that in developed countries, although the findings cannot be generalized. Quality in the public sector is better than in the private sector in many areas, despite spending being substantially less. Quality in public hospitals is resource constrained, and needs greater government investment for improvement, but when resource limitations are not critical, the public sector appears able to deliver equal or better quality than the private sector. Overall similarities in quality between the two sectors suggest the importance of physician training and other factors.


Gliotoxin, a Known Virulence Factor in the Major Human Pathogen Aspergillus fumigatus, Is Also Biosynthesized by Its Nonpathogenic Relative Aspergillus fischeri.
Aspergillus fumigatus is a major opportunistic human pathogen. Multiple traits contribute to A. fumigatus pathogenicity, including its ability to produce specific secondary metabolites, such as gliotoxin. Gliotoxin is known to inhibit the host immune response, and genetic mutants that inactivate gliotoxin biosynthesis (or secondary metabolism in general) attenuate A. fumigatus virulence. The genome of Aspergillus fischeri, a very close nonpathogenic relative of A. fumigatus, contains a biosynthetic gene cluster that is homologous to the A. fumigatus gliotoxin cluster. However, A. fischeri is not known to produce gliotoxin. To gain further insight into the similarities and differences between the major pathogen A. fumigatus and the nonpathogen A. fischeri, we examined whether A. fischeri strain NRRL 181 biosynthesizes gliotoxin and whether the production of secondary metabolites influences the virulence profile of A. fischeri We found that A. fischeri biosynthesizes gliotoxin under the same conditions as A. fumigatus However, whereas loss of laeA, a master regulator of secondary metabolite production (including gliotoxin biosynthesis), has previously been shown to reduce A. fumigatus virulence, we found that laeA loss (and loss of secondary metabolite production) in A. fischeri does not influence its virulence. These results suggest that LaeA-regulated secondary metabolites are virulence factors in the genomic and phenotypic background of the major pathogen A. fumigatus but are much less important in the background of the nonpathogen A. fischeri Understanding the observed spectrum of pathogenicity across closely related pathogenic and nonpathogenic Aspergillus species will require detailed characterization of their biological, chemical, and genomic similarities and differences.IMPORTANCEAspergillus fumigatus is a major opportunistic fungal pathogen of humans, but most of its close relatives are nonpathogenic. Why is that so? This important, yet largely unanswered, question can be addressed by examining how A. fumigatus and its close nonpathogenic relatives are similar or different with respect to virulence-associated traits. We investigated whether Aspergillus fischeri, a nonpathogenic close relative of A. fumigatus, can produce gliotoxin, a mycotoxin known to contribute to A. fumigatus virulence. We discovered that the nonpathogenic A. fischeri produces gliotoxin under the same conditions as those of the major pathogen A. fumigatus However, we also discovered that, in contrast to what has previously been observed in A. fumigatus, the loss of secondary metabolite production in A. fischeri does not alter its virulence. Our results are consistent with the "cards of virulence" model of opportunistic fungal disease, in which the ability to cause disease stems from the combination ("hand") of virulence factors ("cards") but not from individual factors per se.


Third-generation cephalosporin resistant Enterobacteriaceae in neonates and young infants: impact and outcome.
Third-generation cephalosporins resistant Enterobacteriaceae (3GCR-EB) are a major threat in severely ill neonates hospitalized in Neonatal Intensive Care Units. Still, the particular impact of 3GCR-EB on outcomes in the wide neonatal population is not well-appreciated. We aimed to study the impact of 3GCR-EB on the length of hospital stay and mortality of a general population of neonates and young infants. This was a retrospective cohort study of neonates and young infants born in eight Israeli hospitals between 2009 and 2013, with a culture taken within three months after birth that tested positive for Enterobacteriaceae (EB). Data for this study were taken from centralized electronic health records included inpatient, outpatient, socio-demographic, administrative and laboratory information. The main outcomes were length of stay and mortality. The main explanatory variable was an isolation of 3GCR-EB in any bacterial culture taken from a neonate or young infant. Cultures were taken for 31,921 neonates and young infants; 2647 (8.3%) tested positive for EB and 290 (11%) tested positive for 3GCR-EB. Length of stay for those who tested positive was 2.8 times longer (95%CI: 2.70-2.91, p ˂ .001) than patients who tested positive for 3GC-susceptible EB. 3GCR-EB were also associated with increased mortality (OR: 12.06, 95%CI: 4.92-32.29). Neonates with third-generation cephalosporins resistant Enterobacteriaceae had extended hospitalization and increased mortality, which was mostly significant in normal gestational weight newborns.


Urinalysis findings and urinary kidney injury biomarker concentrations.
Urinary biomarkers of kidney injury are presumed to reflect renal tubular damage. However, their concentrations may be influenced by other factors, such as hematuria or pyuria. We sought to examine what non-injury related urinalysis factors are associated with urinary biomarker levels. We examined 714 adults who underwent cardiac surgery in the TRIBE-AKI cohort that did not experience post-operative clinical AKI (patients with serum creatinine change of ≥ 20% were excluded). We examined the association between urinalysis findings and the pre- and first post-operative urinary concentrations of 4 urinary biomarkers: neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), kidney injury molecule-1 (KIM-1), and liver fatty acid binding protein (L-FABP). The presence of leukocyte esterase and nitrites on urinalysis was associated with increased urinary NGAL (R2 0.16, p < 0.001 and R2 0.07, p < 0.001, respectively) in pre-operative samples. Hematuria was associated with increased levels of all 4 biomarkers, with a much stronger association seen in post-operative samples (R2 between 0.02 and 0.21). Dipstick proteinuria concentrations correlated with levels of all 4 urinary biomarkers in pre-operative and post-operative samples (R2 between 0.113 and 0.194 in pre-operative and between 0.122 and 0.322 in post-operative samples). Adjusting the AUC of post-operative AKI for dipstick proteinuria lowered the AUC for all 4 biomarkers at the pre-operative time point and for 2 of the 4 biomarkers at the post-operative time point. Several factors available through urine dipstick testing are associated with increased urinary biomarker concentrations that are independent of clinical kidney injury. Future studies should explore the impact of these factors on the prognostic and diagnostic performance of these AKI biomarkers.


Lattice mold technique for the calculation of crystal nucleation rates.
We present a new simulation method for the calculation of crystal nucleation rates by computer simulation. The method is based on the use of molds to induce crystallization in state points where nucleation is a rare event. The mold is a cluster of potential energy wells placed in the lattice positions of the solid. The method has two distinct steps. In the first one the probability per unit volume of forming a sub-critical crystal cluster in the fluid is computed by means of thermodynamic integration. The thermodynamic route consists in gradually switching on an attractive interaction between the wells and the fluid particles. In the second step, the frequency with which such cluster becomes post-critical is computed in Molecular Dynamics simulations with the mold switched on. We validate our method with a continuous version of the hard sphere potential and with the sodium chloride Tosi-Fumi model. In all studied state points we obtain a good agreement with literature data obtained from other rare event simulation techniques. Our method is quite suitable for the study of both crystal nucleation of arbitrarily complex structures and the competition between different polymorphs in the nucleation stage.


Modulation of astrocytic metabolic phenotype by proinflammatory cytokines.
Astrocytes play an important role in nervous system homeostasis. In particular, they contribute to the regulation of local energy metabolism and to oxidative stress defence. In previous experiments, we showed that long-term treatment with interleukin 1alpha (IL-1alpha) or tumor necrosis factor-alpha (TNFalpha) alone increases glucose utilization in primary culture of mouse astrocytes. In our study, we report that a combination of IL-1beta and TNFalpha exerts a synergistic effect on glucose utilization and markedly modifies the metabolic phenotype of astrocytes. Thus, IL-1beta+TNFalpha treated astrocytes show a marked decrease in glycogen levels, a slight but not significant decrease in lactate release as well as a massive increase in both the pentose phosphate pathway and TCA cycle activities. Glutamate-stimulated glucose utilization and lactate release, a typical feature of astrocyte energy metabolism, are altered after pretreatment with IL-1beta+TNFalpha. As far as mechanisms for oxidative stress defence are concerned, we observed that treatment with IL-1beta+TNFalpha decreases cellular glutathione content and increases glutathione release into the extracellular space while stimulating superoxide anion and nitric oxide production as well as H(2)O(2) release. Interestingly, stimulation of glucose utilization by IL-1beta+TNFalpha is not affected by the antioxidant N-acetyl-L-cysteine, suggesting that cellular stress does not account for this effect. Finally, the effects of cytokines on glucose utilization appear to involve multiple signaling cascades including the phosphoinositide 3-kinase and mitogen-activated protein kinases. Taken together these results establish that a proinflammatory environment such as observed in several neuropathological conditions including Alzheimer's disease, markedly modifies the metabolic phenotype of astrocytes.


Inhibition of contraction of cultured muscle fibers results in increased turnover of myofibrillar proteins but not of intermediate-filament proteins.
Muscle fibers are maintained in culture in a fully contractile state and are relaxed by the addition of 10(-7) M tetrodotoxin (TTX). This toxin binds to muscle membrane Na+- channels, abolishes spontaneous contractions and causes failure of the fiber to accumulate myosin heavy chains. These effects are reversible on removal of TTX. Synthesis and accumulation kinetics have been obtained for myofibrillar and for cytoplasmic filament proteins in normal, active muscle and in TTX-relaxed muscle fibers in culture. In relaxed fibers the synthesis of most proteins remained normal or slightly elevated. However, the accumulation of all myofibrillar proteins examined was markedly inhibited in TTX-treated cultures, whereas the accumulation of cytoplasmic filament proteins was normal or slightly elevated. Myofibrillar proteins examined were alpha-actin, troponin-C, myosin fast light chain 1, myosin fast light chain 2, alpha, beta-tropomyosins and the phosphorylated forms of tropomyosin and fast light chain 2. Cytoplasmic filament proteins studied were vimentin, alpha, beta-desmin and beta, alpha-actin. We also examined the synthesis and accumulation of six unidentified muscle-specific proteins and nine unidentified nonmuscle-specific proteins. Most of these proteins showed a normal accumulation pattern in TTX-relaxed fibers. We concluded that muscle fibers made inactive by TTX display an increased instability of all myofibrillar proteins while cytoplasmic filament proteins and cytoplasmic proteins in general are relatively unaffected. We suggest that TTX interferes, in a manner as yet unidentified, with assembly and normal stability of myofibrils. Decreased assembly and/or increased instability of myofibrils would lead to increased rates of myofibrillar protein degradation.


Coronary artery disease in cardiac transplant patients receiving triple-drug immunosuppressive therapy.
Although triple-drug immunosuppression with cyclosporine, prednisone, and azathioprine has reduced the incidence of acute graft rejection after cardiac transplant, its effect on the development of coronary artery disease is unknown. We have followed up 74 cardiac transplant recipients with yearly coronary angiograms. The probability of acute rejection was 10.8% at 1 year. The incidence of coronary artery disease was 8% at 1 year (six of 74 patients) and 24% at 2 years (11 of 30 patients). In patients who developed posttransplant coronary artery disease, there was a slightly higher, but not statistically significant, incidence of acute graft rejection and pretransplant idiopathic cardiomyopathy. No correlation was found between the incidence of coronary artery disease and recipient age, human leukocyte antigen (HLA) type, hypertension, diabetes, cholesterol level, triglyceride levels, weight gain after transplant, and smoking. These data indicate that triple-drug immunosuppression, despite having produced a significant reduction in the episodes of acute graft rejection, has not decreased the incidence of posttransplant coronary artery disease. Common risk factors for coronary disease and HLA mismatch are probably not important in the pathogenesis of coronary atherosclerosis after cardiac transplantation, whereas the risk for coronary artery disease may be increased by acute graft rejection and pretransplant idiopathic cardiomyopathy.


[Transobturator tapes for stress urinary incontinence treatment: middle-term results].
Minimal invasive surgical treatment of female stress urinary incontinence with suburethral transobturator tapes is completely accepted nowadays. The purpose of this study is to evaluate the efficacy and security of these suburethral tapes, comparing the outside-in to the inside-out approach. We performed a retrospective study of 298 patients with stress urinary incontinence diagnosis that were submitted to transobturator surgery between 2003 and 2006. From these patients 113 women underwent outside-in approach and 185 patients had inside-out approach. The mean-age of the patients was 57.2 +/- 10.3 years-old and 69.1% were in the post-menopausal period. Mean parity was 2.2 +/- 1.1. The outside-in approach was performed more frequently in association with other(s) pelvic floor surgery(ies) (83.2% versus 37.8% for the inside-out technique). The mean follow-up duration was of 14.35 +/- 13.75 months in those patients who had a transobturator outside-in technique and of 11.79 +/- 10.39 months in the inside-out group. In respect to the efficacy, results were identical in the two groups with cure and cure or improvement rates respectively of 76.9% and 92.9% in the outside-in group and of 82.7% and 93.5% in the inside-out group (differences not significative). Regarding mean procedure duration, when it was performed isolated, we had a significant lower duration in patients submitted to inside-out technique (14.77 +/- 5.37 minutes versus 21.21 +/- 7.48 minutes, p < 0.05). The post-operative rates of de novo urge incontinence and mesh erosions were identical in the two groups, however microporous tapes had more erosions than macroporous ones (p < 0.05). The transobturator suburethral tapes are effective and safe for the stress urinary incontinence treatment. We had elevated cure and improvement rates with no significant differences in regard to performed procedure. The inside-out technique is significantly associated to a lower operative duration.


Cannabis and alcohol use, and the developing brain.
Sex hormones and white (and grey) matter in the limbic system, cortex and other brain regions undergo changes during adolescence. Some of these changes include ongoing white matter myelination and sexually dimorphic features in grey and white matter. Adolescence is also a period of vulnerability when many are first exposed to alcohol and cannabis, which appear to influence the developing brain. Neuropsychological studies have provided considerable understanding of the effects of alcohol and cannabis on the brain. Advances in neuroimaging have allowed examination of neuroanatomic changes, metabolic and neurotransmitter activity, and neuronal activation during adolescent brain development and substance use. In this review, we examine major differences in brain development between users and non-users, and recent findings on the influence of cannabis and alcohol on the adolescent brain. We also discuss associations that appear to resolve following short-term abstinence, and attentional deficits that appear to persist. These findings can be useful in guiding earlier educational interventions for adolescents, and clarifying the neural sequelae of early alcohol and cannabis use to the general public.