The Parahox gene Pdx1 is required to maintain positional identity in the adult foregut.
The homeobox gene Pdx1 is a key regulator of pancreas and foregut development. Loss of Pdx1 expression results in pancreas agenesis and impaired development of the gastro-duodenal domain including Brunner's glands. We previously demonstrated a key role for Pdx1 in maintaining the integrity and function of insulin-secreting beta cells in the adult pancreas. In the present study, we aimed to determine if expression of Pdx1 is required to maintain the cellular identity of the gastro-duodenal domain in adult mice. Immunohistological studies were performed in a mouse model in which expression of Pdx1 was conditionally repressed with the doxycycline-responsive tetracycline transactivator system. Mice in which Pdx1 was chronically repressed developed hamartomas in the gastro-duodenal domain. These lesions appeared to arise from ectopic foci of anteriorized cells, consistent with a localised anterior homeotic shift. They emerge with the intercalation of tissue between the anteriorized and normal domains and appear strikingly similar to lesions in the colon of mice heterozygous for another Parahox gene, Cdx2. Continuing expression of Pdx1 into adult life is required to maintain regional cellular identity in the adult foregut, specifically at the gastro-duodenal boundary. Loss of Pdx1 expression leads to anterior transformation and intercalary regeneration of ectopic tissue. We propose a model in which the posterior dominance of classical Hox genes is mirrored by the Parahox genes, providing further evidence of the functional conservation of the Parahox genes. These findings may have implications for further understanding the molecular basis of gastro-duodenal metaplasia and gastro-intestinal transformations such as Barrett's esophagus.


Multilevel examination of minor salivary gland biopsy for Sjogren's syndrome significantly improves diagnostic performance of AECG classification criteria.
The recently observed low reproducibility of focus score (FS) assessment at different section depths in a series of single minor salivary gland biopsies highlighted the need for a standardized protocol of extensive histopathological examination of such biopsies in Sjogren's syndrome. For this purpose, a cumulative focus score (cFS) was evaluated on three slides cut at 200-mum intervals from each of a series of 120 salivary biopsies. The cFS was substituted for the baseline FS in the American-European Consensus Group (AECG) criteria set for Sjogren's syndrome classification, and then test specificity and sensitivity were assessed against clinical patient re-evaluation. Test performances of the AECG classification with the original FS and the score obtained after multilevel examination were statistically compared using receiver operating characteristic (ROC) curve analysis. The diagnostic performance of AECG classification significantly improved when the cFS was entered in the AECG classification; the improvement was mostly due to increased specificity in biopsies with a baseline FS >or= 1 but <2. The assessment of a cFS obtained at three different section levels on minor salivary gland biopsies can be useful especially in biopsies with baseline FSs between 1 and 2.


The physiology of external cardiac massage: high-impulse cardiopulmonary resuscitation.
In intact chronically instrumented dogs, left ventricular dynamics were studied during cardiopulmonary resuscitation (CPR). Electromagnetic flow probes measured cardiac output and coronary blood flow, ultrasonic transducers measured cardiac dimensions, and micromanometers measured left ventricular, right ventricular, aortic, and intrathoracic pressures. The dogs were anesthetized with morphine, intubated, and fibrillated by rapid ventricular pacing. Data were obtained during manual external massage with dogs in the lateral and supine positions. Force of compression was varied from a peak intrathoracic pressure of 10 to 30 mm Hg, and compression rate was varied from 60 to 150/min. Increasing force of compression increased stroke volume up to a peak intrathoracic pressure of approximately 20 mm Hg, beyond which stroke volume remained constant or declined. Stroke volume appeared to result primarily from direct transmission of manual compression force to the heart rather than from positive intrathoracic pressure because peak cardiac or vascular pressures or the change in these pressures were consistently two to four times greater than the corresponding intrathoracic pressures during manual compression. With increasing compression rate, stroke volume remained relatively constant, and total cardiac output increased significantly: 425 +/- 92 ml/min at 60/min, 643 +/- 130 ml/min at 100/min, and 975 +/- 219 ml/min at 150/min (p less than .05). Left ventricular dimensions decreased minimally at higher manual compression rates. In four patients undergoing CPR, systolic and diastolic arterial blood pressure increased with faster compression rates, correlating well with data obtained in the dog. Dynamic coronary blood flow in canine experiments decreased to zero or negative values during compression. Antegrade coronary flow occurred primarily during noncompression periods and seemed to be related to diastolic aortic perfusion pressure; coronary flow at a compression rate of 150/min averaged 75% of control. Therefore stroke volume and coronary blood flow in this canine preparation were maximized with manual chest compression performed with moderate force and brief duration. Increasing rate of compression increased total cardiac output while coronary blood flow was well maintained. Direct cardiac compression appeared to be the major determinant of stroke volume during manual external cardiac massage.


Gastroenterology in the Trent Region in 1992 and a review of changes since 1975.
A survey of all gastroenterologists in the Trent Region (population 4.7 million) showed that in 1992 the number of upper gastrointestinal endoscopies had continued to rise considerably. There were 8.6 per thousand of the population. Colonoscopies had also increased to 1.4 per thousand. There had been a large fall in barium meal examinations to 4.8 per thousand. Barium enema examinations had increased slightly to 5.3 per thousand. The number of endoscopic retrograde cholangiopancreatographies (ERCPs) and variceal sclerotherapies had also increased by 54% and 21% respectively since 1986 and were available in most of the 20 hospitals. Percutaneous endoscopic gastrostomy had been introduced at most hospitals. Emergency endoscopy was readily available at night in only half the hospitals. Fourteen hospitals offered formal open access endoscopy. Intravenous sedation was used routinely in 12 hospitals. The rate for perforation at routine upper gastrointestinal endoscopy was 1 in 4500 and for death was 1 in 20,000. There were serious deficiencies in provision of facilities or staff, or both at 14 hospitals. The total number of gastroenterologists had only increased from 25 to 26 since 1986. Thus endoscopy workload continues to increase and diversify often without adequate staffing, facilities, and funding.


Mechanisms of ethylene biosynthesis and response in plants.
Ethylene is the simplest unsaturated hydrocarbon, yet it has profound effects on plant growth and development, including many agriculturally important phenomena. Analysis of the mechanisms underlying ethylene biosynthesis and signalling have resulted in the elucidation of multistep mechanisms which at first glance appear simple, but in fact represent several levels of control to tightly regulate the level of production and response. Ethylene biosynthesis represents a two-step process that is regulated at both the transcriptional and post-translational levels, thus enabling plants to control the amount of ethylene produced with regard to promotion of responses such as climacteric flower senescence and fruit ripening. Ethylene production subsequently results in activation of the ethylene response, as ethylene accumulation will trigger the ethylene signalling pathway to activate ethylene-dependent transcription for promotion of the response and for resetting the pathway. A more detailed knowledge of the mechanisms underlying biosynthesis and the ethylene response will ultimately enable new approaches to be developed for control of the initiation and progression of ethylene-dependent developmental processes, many of which are of horticultural significance.


Processes that drive the population structuring of Jenynsia lineata (Cyprinidontiformes, Anablepidae) in the La Plata Basin.
The distribution of genetic diversity across a species distribution range is rarely homogeneous, as the genetic structure among populations is related to the degree of isolation among them, such as isolation by distance, isolation by barrier, and isolation by environment. Jenynsia lineata is a small viviparous fish that inhabits a wide range of habitats in South America. To decipher the isolation processes that drive population structuring in J. lineata, we analyzed 221 sequences of the mitochondrial cytochrome c oxidase I gene (COI), from 19 localities. Then, we examined the influence of the three most common types of isolation in order to explain the genetic variation found in this species.Our results revealed a marked structuration, with three groups: (a) La Plata/Desaguadero Rivers (sampling sites across Argentina, Uruguay, and Southern Brazil), (b) Central Argentina, and (c) Northern Argentina. A distance-based redundancy analysis, including the explanatory variables geographical distances, altitude, latitude, and basin, was able to explain up to 65% of the genetic structure. A variance partitioning analysis showed that the two most important variables underlying the structuration in J. lineata were altitude (isolation by environment) and type of basin (isolation by barrier).Our results show that in this species, the processes of population diversification are complex and are not limited to a single mechanism. The processes that play a prominent role in this study could explain the high rate of diversity that characterizes freshwater fish species. And these processes in turn are the basis for possible speciation events.


Evaluation of traumatic dental injuries in a group of Turkish population.
Traumatic dental injuries (TDIs) are significant clinical problems that may have complex etiologies and may cause physical and physiological impairment. The aim of this study was to determine the demographic and clinical characteristics of TDIs in a group of Turkish subjects. The prevalence of TDIs and the pattern of traumatized teeth were studied in relation to age, gender, cause, location, and type of injury. This study was carried out to include all age groups and teeth. TDIs were recorded using the World Health Organization classification modified by Andreasen et al. The prevalence of TDIs was found to be 4.4% (255 injured patients/5800 presented patients), and the injuries were most commonly observed in the age group of 11-20 years. Males were more affected than females (males, 153; females, 102). The most common cause of traumatic injury was falling (68.2%), and the most common place of trauma was outdoors (56.1%). 4, 7% of patients injured because of their systemic diseases. The most frequently injured teeth were the upper central incisors (primary teeth, 64.5%; permanent teeth, 72.5%), and the most common type of dental injury was uncomplicated crown fracture in both primary (63%) and permanent dentition (47%). In the surveyed population, the prevalence of TDIs was found to be low. However, the teacher and family of adolescents and the caregivers of patients with some systemic diseases like epilepsy should be warned about TDIs.


Decreases in bone mineral density at cortical and trabecular sites in the tibia and femur during the first year of spinal cord injury.
Disuse osteoporosis occurs in response to long-term immobilization. Spinal cord injury (SCI) leads to a form of disuse osteoporosis that only affects the paralyzed limbs. High rates of bone resorption after injury are evident from decreases in bone mineral content (BMC), which in the past have been attributed in the main to loss of trabecular bone in the epiphyses and cortical thinning in the shaft through endocortical resorption. Patients with motor-complete SCI recruited from the Queen Elizabeth National Spinal Injuries Unit (Glasgow, UK) were scanned within 5weeks of injury (baseline) using peripheral Quantitative Computed Tomography (pQCT). Unilateral scans of the tibia, femur and radius provided separate estimates of trabecular and cortical bone parameters in the epiphyses and diaphyses, respectively. Using repeat pQCT scans at 4, 8 and 12months post-injury, changes in BMC, bone mineral density (BMD) and cross-sectional area (CSA) of the bone were quantified. Twenty-six subjects (5 female, 21 male) with SCI (12 paraplegic, 14 tetraplegic), ranging from 16 to 76years old, were enrolled onto the study. Repeated-measures analyses showed a significant effect of time since injury on key bone parameters at the epiphyses of the tibia and femur (BMC, total BMD, trabecular BMD) and their diaphyses (BMC, cortical BMD, cortical CSA). There was no significant effect of gender or age on key outcome measures, but there was a tendency for the female subjects to experience greater decreases in cortical BMD. The decreases in cortical BMD in the tibia and femur were found to be statistically significant in both men and women. By carrying out repeat pQCT scans at four-monthly intervals, this study provides a uniquely detailed description of the cortical bone changes that occur alongside trabecular bone changes in the first year of complete SCI. Significant decreases in BMD were recorded in both the cortical and trabecular bone compartments of the tibia and femur throughout the first year of injury. This study provides evidence for the need for targeted early intervention to preserve bone mass within this patient group.


Children with intestinal failure are at risk for psychopathology and trauma.
To assess psychopathology and medical traumatic stress in children with intestinal failure (IF) and identify associated risk factors. Two-center study, performed from September 2019 until April 2022 (partly during COVID-19 pandemic), including children (1.5-17y) with IF, dependent on parenteral nutrition (PN) or weaned off PN, treated by a multidisciplinary IF-team. Psychopathology in children was evaluated with a semi-structured interview assessing psychiatric classifications and validated questionnaires assessing emotional (internalizing) and behavioral (externalizing) problems. Medical traumatic stress was assessed with a validated questionnaire. Problem scores were compared with normative data. Associations between clinical characteristics and outcomes were analyzed with linear regression analyses. Forty-one (of 111 eligible) children were included (median age 8.9y (IQR 5.5-11.8), 54% female, 73% born preterm). Median PN-duration was 17.3 months (IQR 6.9-54.0); 17 children (41%) were still PN-dependent. One third of the children met criteria for at least one psychiatric classification (compared with 14% in age-matched general population). Anxiety disorders and attention deficit hyperactivity disorder were most common. In school-aged children (n=29, 6-17y), significantly increased emotional problems were consistently reported by children (p=0.011), parents (p<0.001) and teachers (p=0.004). In preschool children (n=12, 1.5-5y), no significant differences with normative data were found. Subclinical or clinical emotional problems were reported in 19 children (46%). Medical traumatic stress was present in 14%, and 22% of children had received psychological help for trauma before. Lower gastrointestinal related quality of life was associated with more emotional problems, but not PN-duration. Children with IF, particularly school-aged children, are at risk for psychological problems which is reflected by the high rate of received psychotherapy and the high rate of emotional problems and psychiatric classifications.


Effects of Caloric Restriction with Protein Supplementation on Plasma Protein Profiles in Middle-Aged Women with Metabolic Syndrome-A Preliminary Open Study.
Clinical studies have demonstrated that higher protein intake based on caloric restriction (CR) alleviates metabolic abnormalities. However, no study has examined the effects of plasma protein profiles on caloric restriction with protein supplementation (CRPS) in metabolic syndrome (MetS). Therefore, using a proteomic perspective, this pilot study investigated whether CRPS ameliorated metabolic abnormalities associated with MetS in middle-aged women. Plasma samples of middle-aged women with MetS in CR (n = 7) and CRPS (n = 6) groups for a 12-week intervention were obtained and their protein profiles were analysed. Briefly, blood samples from qualified participants were drawn before and after the dietary treatment. Anthropometric, clinical, and biochemical variables were measured and correlated with plasma proteomics. In results, we found that body mass index, total body fat, and fasting blood glucose decreased significantly after the interventions but were not different between the CR and CRPS groups. After liquid chromatography⁻tandem mass spectrometry analysis, the relative plasma levels of alpha-2-macroglobulin (A2M), C4b-binding protein alpha chain (C4BPA), complement C1r subcomponent-like protein (C1RL), complement component C6 (C6), complement component C8 gamma chain (C8G), and vitamin K-dependent protein S (PROS) were significantly different between the CRPS and CR groups. These proteins are involved in inflammation, the immune system, and coagulation responses. Moreover, blood low-density lipoprotein cholesterol levels were significantly and positively correlated with C6 plasma levels in both groups. These findings suggest that CRPS improves inflammatory responses in middle-aged women with MetS. Specific plasma protein expression (i.e., A2M, C4BPA, C1RL, C6, C8G, and PROS) associated with the complement system was highly correlated with fasting blood glucose (FBG), blood lipids (BLs), and body fat.


Automated diabetic macular edema (DME) grading system using DWT, DCT Features and maculopathy index.
The cause of diabetic macular edema (DME) is due to prolonged and uncontrolled diabetes mellitus (DM) which affects the vision of diabetic subjects. DME is graded based on the exudate location from the macula. It is clinically diagnosed using fundus images which is tedious and time-consuming. Regular eye screening and subsequent treatment may prevent the vision loss. Hence, in this work, a hybrid system based on Radon transform (RT), discrete wavelet transform (DWT) and discrete cosine transform (DCT) are proposed for an automated detection of DME. The fundus images are subjected to RT to obtain sinograms and DWT is applied on these sinograms to extract wavelet coefficients (approximate, horizontal, vertical and diagonal). DCT is applied on approximate coefficients to obtain 2D-DCT coefficients. Further, these coefficients are converted into 1D vector by arranging the coefficients in zig-zag manner. This 1D signal is subjected to locality sensitive discriminant analysis (LSDA). Finally, various supervised classifiers are used to classify the three classes using significant features. Our proposed technique yielded a classification accuracy of 100% and 97.01% using two and seven significant features for private and public (MESSIDOR) databases respectively. Also, a maculopathy index is formulated with two significant parameters to discriminate the three groups distinctly using a single integer. Hence, our obtained results suggest that this system can be used as an eye screening tool for diabetic subjects for DME.


Aotus lemurinus griseimembra monkeys: a suitable model for Plasmodium vivax sporozoite infection.
This study describes a successful Plasmodium vivax sporozoite infection in Aotus lemurinus griseimembra. Twenty-eight naive or previously infected monkeys, either splenectomized or spleen intact, were inoculated intravenously or subcutaneously with Plasmodium vivax sporozoites of the Salvador I strain or with two wild isolates (VCC-4 and VCC-5; Vivax-Cali-Colombia). The monkeys were successfully infected regardless of the parasite strain, spleen presence, or inoculation route and showed prepatent periods that ranged from 16 to 89 days. Only one monkey inoculated intravenously failed to develop parasitemia. Since immune protection against malaria pre-erythrocytic forms is mediated by both helper and cytolytic T cells that may home in the spleen and P. vivax cultures are not yet available; the use of spleen-intact A. lemurinus griseimembra, susceptible to both adapted and non-adapted strains of P. vivax sporozoites, is a valuable model for evaluation of pre-erythrocytic vaccine candidates.


New developments in cognitive behavioral therapy as the first-line treatment of insomnia.
Insomnia is the most common sleep disorder. Psychological, behavioral, and biological factors are implicated in the development and maintenance of insomnia as a disorder, although the etiology of insomnia remains under investigation, as it is still not fully understood. Cognitive behavioral therapy for insomnia (CBTI) is a treatment for insomnia that is grounded in the science of behavior change, psychological theories, and the science of sleep. There is strong empirical evidence that CBTI is effective. Recognition of CBTI as the first-line treatment for chronic insomnia (National Institutes of Health consensus, British Medical Association) was based largely on evidence of its efficacy in primary insomnia. The aim of this article is to provide background information and review recent developments in CBTI, focusing on three domains: promising data on the use of CBTI when insomnia is experienced in the presence of comorbid conditions, new data on the use of CBTI as maintenance therapy, and emerging data on the delivery of CBTI through the use of technology and in primary care settings.


Aldosterone-mediated apical targeting of ENaC subunits is blunted in rats with streptozotocin-induced diabetes mellitus.
Diabetes mellitus (DM) is associated with a significant polyuria and natriuesis as well as increased plasma aldosterone and anti-diuretic hormone arginine vasopressin (AVP). This study aimed to determine whether diabetic kidneys compensate for the urinary sodium and water losses by increasing apical targeting of epithelial sodium channel (ENaC) subunits and aquaporin-2 (AQP2) in the collecting duct, in addition to the previously observed changes in ENaC subunit protein expression in different kidney zones. Female rats were investigated 2 weeks after induction of DM by streptozotocin administration. Kidneys were examined by immunohistochemisty and semiquantitative immunoblotting. We demonstrated that the protein expression of renal AQP2, Ser-256 phosphorylated AQP2, AQP3, beta- and gamma-ENaC (but not alpha-ENaC) increased consistently with an increased AVP response. In contrast, there were no significant changes in the relative apical targeting of beta-, gamma- and alpha-ENaC, and the shift in the molecular weight of gamma-ENaC from 85 kDa to 70 kDa was not observed despite increased plasma aldosterone levels. These results were supported by changes in the functional data showing increased solute-free water reabsorption, increased fractional excretion of sodium and an unchanged ratio of potassium to sodium in the urine. The data demonstrate that diabetic kidneys have a reduced sensitivity to the anti-natriuretic action of elevated plasma aldosterone levels with no relative increase in ENaC subunit apical targeting, whereas there is increased expression of beta- and gamma-ENaC, which alone may play a role in the increased sodium reabsorption in the kidney in DM.


Language in tuberculosis services: can we change to patient-centred terminology and stop the paradigm of blaming the patients?
The words 'defaulter', 'suspect' and 'control' have been part of the language of tuberculosis (TB) services for many decades, and they continue to be used in international guidelines and in published literature. From a patient perspective, it is our opinion that these terms are at best inappropriate, coercive and disempowering, and at worst they could be perceived as judgmental and criminalising, tending to place the blame of the disease or responsibility for adverse treatment outcomes on one side-that of the patients. In this article, which brings together a wide range of authors and institutions from Africa, Asia, Latin America, Europe and the Pacific, we discuss the use of the words 'defaulter', 'suspect' and 'control' and argue why it is detrimental to continue using them in the context of TB. We propose that 'defaulter' be replaced with 'person lost to follow-up'; that 'TB suspect' be replaced by 'person with presumptive TB' or 'person to be evaluated for TB'; and that the term 'control' be replaced with 'prevention and care' or simply deleted. These terms are non-judgmental and patient-centred. We appeal to the global Stop TB Partnership to lead discussions on this issue and to make concrete steps towards changing the current paradigm.


Top-down fabrication meets bottom-up synthesis for nanoelectronic barcoding of microparticles.
Traditional optical and plasmonic techniques for barcoding of micro-particles for multiplexed bioassays are generally high in throughput, however bulky instrumentation is often required for performing readout. Electrical impedance based detection allows for ultra-compact instrumentation footprint necessary for wearable devices, however to date, the lack of ability to electronically barcode micro-particles has been a long standing bottleneck towards enabling multiplexed electronic biomarker assays. Nanoelectronic barcoding, which to the best of our knowledge is the first impedance based solution for micro-particle barcoding, works by forming tunable nano-capacitors on the surface of micro-spheres, effectively modulating the frequency dependent dielectric properties of the spheres allowing one bead barcode to be distinguished from another. Nanoelectronic barcoding uses a well-known, but unexplored electromagnetic phenomenon of micro-particles: the Clausius-Mossotti (CM) factor spectrum of a Janus particle (JP) shifts depending on the zeta (wall) potential of the metallic half of the microsphere, and the fact that the complex impedance spectrum of a particle directly corresponds to the CM factor spectrum. A one-to-one correspondence will be established between each biomarker and the corresponding engineered microsphere. This transformative new method for barcoding will enable a new class of handheld and wearable biosensors capable of multiplexed continuous temporal bio-monitoring. The proposed nano-electronically barcoded particles utilize both bottom-up synthesis and top-down fabrication to enable precisely engineered frequency dependent dielectric signatures. Multi-frequency lock-in measurements of the complex impedance, in conjunction with multi-variate analysis of impedance data, allows for particle differentiation using a fully functional ultra-compact electronic detector.


A cognitive approach for design of a multimedia informed consent video and website in pediatric research.
Poor participant comprehension of research procedures following the conventional face-to-face consent process for biomedical research is common. We describe the development of a multimedia informed consent video and website that incorporates cognitive strategies to enhance comprehension of study related material directed to parents and adolescents. A multidisciplinary team was assembled for development of the video and website that included human subjects professionals; psychologist researchers; institutional video and web developers; bioinformaticians and programmers; and parent and adolescent stakeholders. Five learning strategies that included Sensory-Modality view, Coherence, Signaling, Redundancy, and Personalization were integrated into a 15-min video and website material that describes a clinical research trial. A diverse team collaborated extensively over 15months to design and build a multimedia platform for obtaining parental permission and adolescent assent for participant in as asthma clinical trial. Examples of the learning principles included, having a narrator describe what was being viewed on the video (sensory-modality); eliminating unnecessary text and graphics (coherence); having the initial portion of the video explain the sections of the video to be viewed (signaling); avoiding simultaneous presentation of text and graphics (redundancy); and having a consistent narrator throughout the video (personalization). Existing conventional and multimedia processes for obtaining research informed consent have not actively incorporated basic principles of human cognition and learning in the design and implementation of these processes. The present paper illustrates how this can be achieved, setting the stage for rigorous evaluation of potential benefits such as improved comprehension, satisfaction with the consent process, and completion of research objectives. New consent strategies that have an integrated cognitive approach need to be developed and tested in controlled trials.


Antecedent hyperglycemia is associated with an increased risk of neutropenic infections during bone marrow transplantation.
To use bone marrow transplantation (BMT) as a model for testing the association between hyperglycemia and infection. This cohort study included 382 adults (6.5% with diabetes) who had no evidence of infection before neutropenia during BMT. Mean glucose was calculated from central laboratory and bedside measurements taken before neutropenia; the primary outcome was neutropenic infections. Eighty-four patients (22%) developed at least one neutropenic infection, including 51 patients (13%) with bloodstream infections. In patients who did not receive glucocorticoids during neutropenia, each 10 mg/dl increase in mean preneutropenia glucose was associated with an odds ratio of 1.08 (95% CI 0.98-1.19) (P = 0.14) for any infection and 1.15 (1.03-1.28) (P = 0.01) for bloodstream infections, after adjusting for age, sex, race, year, cancer diagnosis, transplant type, and total glucocorticoid dose before neutropenia. In those who received glucocorticoids during neutropenia (n = 71), the adjusted odds ratio associated with a 10 mg/dl increase in mean glucose was 1.21 (1.09-1.34) (P < 0.0001) for any infection and 1.24 (1.11-1.38) (P < 0.0001) for bloodstream infections. There was no association between mean glycemia and long length of hospital stay, critical status designation, or mortality. In a BMT population highly susceptible to infection, there was a continuous positive association between mean antecedent glycemia and later infection risk, particularly in patients who received glucocorticoids while neutropenic. Tight glycemic control during BMT and glucocorticoid treatment may reduce infections.


Cost-effectiveness analysis of endoscopic tympanoplasty versus microscopic tympanoplasty for chronic otitis media in Taiwan.
Health care systems and physicians need to conform to budgets and streamline resources to provide cost-effective quality care. Although endoscopic tympanoplasty (ET) has been performed for decades, no studies on the cost-effectiveness of ET and microscopic tympanoplasty (MT) for treating chronic otitis media have been published. The present study aimed to compare the cost-effectiveness of ET and MT for treating chronic otitis media. This study was performed using a Cohort-style Markov decision-tree economic model with a 30-year time horizon. The economic perspective was that of a third-party payer (Taiwan National Health Insurance System). Two treatment strategies were compared, namely ET and MT. The primary outcome was the incremental cost per quality-adjusted life year (QALY). Probabilities were obtained from meta-analyses. Costs were obtained from the published literature and Taiwan National Health Insurance System database. Multiple sensitivity analyses were performed to account for data uncertainty. The reference case revealed that the total cost of ET was $NT 20,901 for 17.08 QALY per patient. By contrast, the total cost of MT was $NT 21,171 for 17.15 QALY per patient. The incremental cost effectiveness ratio for ET versus that of MT was $NT 3703 per QALY. The cost-effectiveness acceptability curve indicated that ET was comparable to MT at a willingness-to-pay threshold of larger than $NT 35,000 per QALY. This cost-effectiveness analysis indicates that ET is comparable to MT for treating chronic otitis media in Taiwan. This result provides the latest information for physicians, the government, and third-party payers to select proper clinical practice.


Visual search performance is predicted by both prestimulus and poststimulus electrical brain activity.
An individual's performance on cognitive and perceptual tasks varies considerably across time and circumstances. We investigated neural mechanisms underlying such performance variability using regression-based analyses to examine trial-by-trial relationships between response times (RTs) and different facets of electrical brain activity. Thirteen participants trained five days on a color-popout visual-search task, with EEG recorded on days one and five. The task was to find a color-popout target ellipse in a briefly presented array of ellipses and discriminate its orientation. Later within a session, better preparatory attention (reflected by less prestimulus Alpha-band oscillatory activity) and better poststimulus early visual responses (reflected by larger sensory N1 waves) correlated with faster RTs. However, N1 amplitudes decreased by half throughout each session, suggesting adoption of a more efficient search strategy within a session. Additionally, fast RTs were preceded by earlier and larger lateralized N2pc waves, reflecting faster and stronger attentional orienting to the targets. Finally, SPCN waves associated with target-orientation discrimination were smaller for fast RTs in the first but not the fifth session, suggesting optimization with practice. Collectively, these results delineate variations in visual search processes that change over an experimental session, while also pointing to cortical mechanisms underlying performance in visual search.


Speech: relation of nonfluency to information value.
In spontaneous speech and oral reading of stutterers, likelihood of nonfluency occurring on any word is not related to the amount of information carried by that word. The same is true for spontaneous speech of normal speakers. Previous contrary findings can be accounted for by the relationship between information value and word length.


Mutation analysis of AMP-activated protein kinase subunits in inherited cardiomyopathies: implications for kinase function and disease pathogenesis.
Familial hypertrophic cardiomyopathy (HCM) has been defined as a disease of the cardiac sarcomere, although sarcomeric protein mutations are not found in one third of cases. We have recently shown that HCM associated with Wolff-Parkinson-White syndrome (WPW) and conduction disease can be caused by mutations in PRKAG2, which encodes the gamma2 subunit of AMPK, an enzyme central to cellular energy homeostasis. AMPK is a heterotrimer composed of one catalytic subunit (alpha) and two regulatory subunits (beta and gamma). Seven known genes encode the subunit isoforms (alpha1, alpha2, beta1, beta2, gamma1, gamma2, gamma3) and all are expressed in the heart. To better understand the role of AMPK mutations in HCM/WPW and other inherited cardiomyophathies, all 7 subunit genes were screened for mutations in a panel of probands: 3 with HCM/WPW, 4 with DCM/WPW, 38 with HCM alone (in whom contractile protein mutations had not been found) and 13 with DCM alone. In total, 73 amplimers were screened in the 58 probands and a number of polymorphisms, including non-conservative substitutions, were identified. However, no further disease-causing mutations were found in any AMPK subunit gene. These results indicate that HCM with WPW is a distinct, but genetically heterogeneous, condition caused by mutations in PRKAG2 and in an unknown gene or genes, not involved in the AMPK complex. Mutations in PRKAG2 appear to specifically cause HCM with WPW and conduction disease, and not other inherited cardiomyopathies. As deleterious alleles were not found in other AMPK subunit isoforms, the mutations affecting PRKAG2 are likely to confer a specific alteration of AMPK function of particular importance in the myocardium.


Speed control in quadrupedal locomotion: principles of limb coordination in the dog.
Long distance, overground locomotion in the dog was observed and analyzed using the two dimensional gait diagram method. Though the velocity of locomotion chosen by the animals over a 1,000 m course varied, a preferred speed generally emerged and was used during most of the experiment. This animal-specific preferred overground velocity was strongly correlated with the animal's limb lengths, and corresponded to the minimum observed swinging velocity of the limbs. Changes in the pattern of limb coordination during three-limb locomotion were also investigated. Depending upon which limb was restrained, dogs used trot-like or gallop-like gaits which exhibited the same temporal and spatial phase differences as were observed during normal locomotion. However, stride length and swing-stance durations were increased relative to those observed in four legged locomotion. Animals with an additional 2 kg weight trotted slowly, but no significant changes in limb movement parameters were found. Locomotion studies performed in darkness resulted in an immediate switch from asymmetrical (galloping) to symmetrical gaits (walking and trotting). These gaits allow for precise foot placement.


Topical application of tranexamic acid for the reduction of bleeding.
Intravenous tranexamic acid reduces bleeding in surgery, however, its effect on the risk of thromboembolic events is uncertain and an increased risk remains a theoretical concern. Because there is less systemic absorption following topical administration, the direct application of tranexamic acid to the bleeding surface has the potential to reduce bleeding with minimal systemic effects. To assess the effects of the topical administration of tranexamic acid in the control of bleeding. We searched the Cochrane Injuries Group Specialised Register; Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library; Ovid MEDLINE®, Ovid MEDLINE® In-Process & Other Non-Indexed Citations, Ovid MEDLINE® Daily and Ovid OLDMEDLINE®; Embase Classic + Embase (OvidSP); PubMed and ISI Web of Science (including Science Citation Index Expanded and Social Science Citation Index (SCI-EXPANDED & CPCI-S)). We also searched online trials registers to identify ongoing or unpublished trials. The search was run on the 31st May 2013. Randomised controlled trials comparing topical tranexamic acid with no topical tranexamic acid or placebo in bleeding patients. Two authors examined the titles and abstracts of citations from the electronic databases for eligibility. Two authors extracted the data and assessed the risk of bias for each trial. Outcome measures of interest were blood loss, mortality, thromboembolic events (myocardial infarction, stroke, deep vein thrombosis and pulmonary embolism) and receipt of a blood transfusion. We included 29 trials involving 2612 participants. Twenty-eight trials involved patients undergoing surgery and one trial involved patients with epistaxis (nosebleed). Tranexamic acid (TXA) reduced blood loss by 29% (pooled ratio 0.71, 95% confidence interval (CI) 0.69 to 0.72; P < 0.0001). There was uncertainty regarding the effect on death (risk ratio (RR) 0.28, 95% CI 0.06 to 1.34; P = 0.11), myocardial infarction (RR 0.33, 95% CI 0.04 to 3.08; P = 0.33), stroke (RR 0.33, 95% CI 0.01 to 7.96; P = 0.49), deep vein thrombosis (RR 0.69, 95% CI 0.31 to 1.57; P = 0.38) and pulmonary embolism (RR 0.52, 95% CI 0.09 to 3.15; P = 0.48). TXA reduced the risk of receiving a blood transfusion by a relative 45% (RR 0.55, 95% CI 0.55 to 0.46; P < 0.0001). There was substantial statistical heterogeneity between trials for the blood loss and blood transfusion outcomes. There is reliable evidence that topical application of tranexamic acid reduces bleeding and blood transfusion in surgical patients, however the effect on the risk of thromboembolic events is uncertain. The effects of topical tranexamic acid in patients with bleeding from non-surgical causes has yet to be reliably assessed. Further high-quality trials are warranted to resolve these uncertainties before topical tranexamic acid can be recommended for routine use.


Microarray analysis of Lyn-deficient B cells reveals germinal center-associated nuclear protein and other genes associated with the lymphoid germinal center.
Lyn is the only member of the Src family expressed in DT40 B cells, which provide a unique model to study the singular contribution of this protein tyrosine kinase (PTK) family to cell signaling. In these cells, gene ablation of Lyn leads to defective B cell receptor signaling. Complementary DNA array analysis of Lyn-deficient DT40 cells shows that the absence of Lyn leads to down-regulation of numerous genes encoding proteins involved in B cell receptor signaling, proliferation, control of transcription, immunity/inflammation response, and cytoskeletal organization. Most of these expression changes have not been previously associated with Lyn PTK signaling. They include alterations in mRNA levels of germinal center-associated nuclear protein (germinal center-associated DNA primase) (GANP), CD74, CD22, NF-kappaB, elongation factor 1alpha, CD79b, octamer binding factor 1, Ig H chain, stathmin, and gamma-actin. Changes in GANP expression were also confirmed in Lyn-deficient mice, suggesting that Lyn PTK has a unique function not compensated for by other Src kinases. Because Lyn-deficient mice have impaired development of germinal centers in spleen, the decreased expression of GANP in the Lyn-deficient DT40 cell line and Lyn-deficient mice suggests that Lyn controls the formation and proliferation of germinal centers via GANP. GANP promoter activity was higher in wild-type vs Lyn-deficient cells. Mutation of the PU.1 binding site reduced activity in wild-type cells and had no effect in Lyn-deficient cells. The presence of Lyn enhanced PU.1 expression in a Northern blot. Thus, the following new signaling pathway has been described: Lyn-->PU.1-->GANP.


Cytokine Levels in Inner Ear Fluid of Young and Aged Mice as Molecular Biomarkers of Noise-Induced Hearing Loss.
Sensorineural hearing loss (SNHL) is the most common sensory deficit worldwide, frequently caused by noise trauma and aging, with inflammation being implicated in both pathologies. Here, we provide the first direct measurements of proinflammatory cytokines in inner ear fluid, perilymph, of adolescent and 2-year-old mice. The perilymph of adolescent mice exposed to the noise intensity resulting in permanent auditory threshold elevations had significantly increased levels of IL-6, TNF-α, and CXCL1 6 h after exposure, with CXCL1 levels being most elevated (19.3 ± 6.2 fold). We next provide the first immunohistochemical localization of CXCL1 in specific cochlear supporting cells, and its presumed receptor, Duffy antigen receptor for chemokines (DARC), in hair cells and spiral ganglion neurons. Our results demonstrate the feasibility of molecular diagnostics of SNHL using only 0.5 μL of perilymph, and motivate future sub-μL based diagnostics of human SNHL based on liquid biopsy of the inner ear to guide therapy, promote hearing protection, and monitor response to treatment.


Type I (insulin dependent) diabetes: a disease of slow clinical onset?
Type I (insulin dependent) diabetes is usually believed to present acutely and it is assumed that metabolic decompensation is sudden. In a prospective family study, however, 10 of 13 subjects developing the disease showed progressive or intermittent development of hyperglycaemia over many months and the others had non-specific symptoms over a long period. All were first degree relatives of a child with type I diabetes; 10 were siblings (aged 5-24) and three were parents (aged 45-58). All possessed HLA-DR4 or DR3, or both, and all but two had been positive for islet cell antibodies for six to 86 months before diagnosis. Ten had non-specific symptoms for two to 14 months before the onset of thirst and polyuria; one remained asymptomatic even when insulin became necessary. Six subjects had an oral glucose tolerance test before clinical onset, of whom five were diabetic by World Health Organisation criteria four, four, six, seven, and 21 months before insulin was needed. Nine showed random blood glucose concentrations above the 97.5th centile (6.3 mmol/l) six to 34 months (median 12) before diagnosis. Two others had a glucose tolerance test result compatible with diabetes but had not reached the stage of needing insulin. Hyperglycaemia is often of insidious onset in type I diabetes, even in children and young adults. Diagnosis will inevitably be late if considered only when acute symptoms of thirst and polyuria develop.


The Contribution of Auditory and Cognitive Factors to Intelligibility of Words and Sentences in Noise.
Understanding the causes for speech-in-noise (SiN) perception difficulties is complex, and is made even more difficult by the fact that listening situations can vary widely in target and background sounds. While there is general agreement that both auditory and cognitive factors are important, their exact relationship to SiN perception across various listening situations remains unclear. This study manipulated the characteristics of the listening situation in two ways: first, target stimuli were either isolated words, or words heard in the context of low- (LP) and high-predictability (HP) sentences; second, the background sound, speech-modulated noise, was presented at two signal-to-noise ratios. Speech intelligibility was measured for 30 older listeners (aged 62-84) with age-normal hearing and related to individual differences in cognition (working memory, inhibition and linguistic skills) and hearing (PTA(0.25-8 kHz) and temporal processing). The results showed that while the effect of hearing thresholds on intelligibility was rather uniform, the influence of cognitive abilities was more specific to a certain listening situation. By revealing a complex picture of relationships between intelligibility and cognition, these results may help us understand some of the inconsistencies in the literature as regards cognitive contributions to speech perception.


Naegleria: another pathogenic ameba studies in germfree guinea pigs.
Free-living amebas of the genus Naegleria, of world-wide distribution and long considered harmless, have been linked etiologically with 57 fatal cases of primary amebic meningoencephalitis during the last decade. Naegleria from cultures derived from one of these fatal cases in Richmond, Virginia, have been inoculated intranasally, intraorally, into the conjunctival sac near the inner canthus of the eyes, and into induced skin lesions in adult germfree guinea pigs. Of 33 animals inoculated intranasally with 18 to 31 amebas, 31 developed a fatal encephalitis. There was considerable destruction of tissues of the cerebellum and the cerebrum and including the olfactory lobes. The meninges were involved to varying degrees in most of the animals. None of the animals inoculated by the three other routes developed either symptoms or lesions.


Ribosomal multilocus sequence typing: universal characterization of bacteria from domain to strain.
No single genealogical reconstruction or typing method currently encompasses all levels of bacterial diversity, from domain to strain. We propose ribosomal multilocus sequence typing (rMLST), an approach which indexes variation of the 53 genes encoding the bacterial ribosome protein subunits (rps genes), as a means of integrating microbial genealogy and typing. As with multilocus sequence typing (MLST), rMLST employs curated reference sequences to identify gene variants efficiently and rapidly. The rps loci are ideal targets for a universal characterization scheme as they are: (i) present in all bacteria; (ii) distributed around the chromosome; and (iii) encode proteins which are under stabilizing selection for functional conservation. Collectively, the rps loci exhibit variation that resolves bacteria into groups at all taxonomic and most typing levels, providing significantly more resolution than 16S small subunit rRNA gene phylogenies. A web-accessible expandable database, comprising whole-genome data from more than 1900 bacterial isolates, including 28 draft genomes assembled de novo from the European Bioinformatics Institute (EBI) sequence read archive, has been assembled. The rps gene variation catalogued in this database permits rapid and computationally non-intensive identification of the phylogenetic position of any bacterial sequence at the domain, phylum, class, order, family, genus, species and strain levels. The groupings generated with rMLST data are consistent with current nomenclature schemes and independent of the clustering algorithm used. This approach is applicable to the other domains of life, potentially providing a rational and universal approach to the classification of life that is based on one of its fundamental features, the translation mechanism.


Temporal Dermoid Cyst with Unusual Imaging Appearance: Case Report.
Intracranial dermoid cysts are benign, slow growing tumors derived from ectopic inclusions of epithelial cells during closure of neural tube. These lesions, accounting for less than 1% of intracranial tumors, have characteristic computed tomography (CT) and magnetic resonance imaging (MRI) appearances that generally permits preoperative diagnosis. However, the radiologic features are uncommon and the cyst can be easily misdiagnosed with other tumors in rare cases. Herein, we report a case of a left temporoparietal dermoid cyst in a 48-year-old woman that was peroperatively and histopathologically proven but not advocated on CT and MRI. Clinical, radiological and histopathological features of a dermoid cyst are reviewed.


Aggravating genetic interactions allow a solution to redundancy in a bacterial pathogen.
Interactions between hosts and pathogens are complex, so understanding the events that govern these interactions requires the analysis of molecular mechanisms operating in both organisms. Many pathogens use multiple strategies to target a single event in the disease process, confounding the identification of the important determinants of virulence. We developed a genetic screening strategy called insertional mutagenesis and depletion (iMAD) that combines bacterial mutagenesis and RNA interference, to systematically dissect the interplay between a pathogen and its host. We used this technique to resolve the network of proteins secreted by the bacterium Legionella pneumophila to promote intracellular growth, a critical determinant of pathogenicity of this organism. This strategy is broadly applicable, allowing the dissection of any interface between two organisms involving numerous interactions.


Gene expression profiling of peripheral blood cells for early detection of breast cancer.
Early detection of breast cancer is key to successful treatment and patient survival. We have previously reported the potential use of gene expression profiling of peripheral blood cells for early detection of breast cancer. The aim of the present study was to refine these findings using a larger sample size and a commercially available microarray platform. Blood samples were collected from 121 females referred for diagnostic mammography following an initial suspicious screening mammogram. Diagnostic work-up revealed that 67 of these women had breast cancer while 54 had no malignant disease. Additionally, nine samples from six healthy female controls were included. Gene expression analyses were conducted using high density oligonucleotide microarrays. Partial Least Squares Regression (PLSR) was used for model building while a leave-one-out (LOO) double cross validation approach was used to identify predictors and estimate their prediction efficiency. A set of 738 probes that discriminated breast cancer and non-breast cancer samples was identified. By cross validation we achieved an estimated prediction accuracy of 79.5% with a sensitivity of 80.6% and a specificity of 78.3%. The genes deregulated in blood of breast cancer patients are related to functional processes such as defense response, translation, and various metabolic processes, such as lipid- and steroid metabolism. We have identified a gene signature in whole blood that classifies breast cancer patients and healthy women with good accuracy supporting our previous findings.


Evaluation of gellan gum fluid gels as modified release oral liquids.
Oral liquids are often preferred for drug administration to patients for whom swallowing is difficult, however, formulating modified release versions can be challenging. A potential route to achieve modified release in oral liquids is by using fluid (sheared) gels formed by introducing a shear field during gelation in gel-forming biopolymers. These fluid gels can act as pourable viscoelastic fluids but retain true gel micro/nano structure. Here, we have demonstrated that fluid gels have potential as paediatric oral liquids preventing release of ibuprofen in simulated gastric fluid. Subsequent release at pH 7.4 was affected by the duration of exposure and magnitude of acid pH with a linear relationship between onset of release and the preceding acidic exposure duration. Delayed release was a result of increasing gel stiffness, a consequence of the acidity of the initial release media and exposure time. A much faster release rate was measured when exposure time in acid was 10 min compared with 60 min. This study highlights the potential to design fluid gels that are tuned to have a specified stiffness at a particular pH and exposure time. This could enable the preparation oral liquids with modified release behaviour.


Epitranscriptome machinery in Trypanosomatids: New players on the table?
Trypanosoma and Leishmania parasites cause devastating tropical diseases resulting in serious global health consequences. These organisms have complex life cycles with mammalian hosts and insect vectors. The parasites must, therefore, survive in different environments, demanding rapid physiological and metabolic changes. These responses depend upon regulation of gene expression, which primarily occurs posttranscriptionally. Altering the composition or conformation of RNA through nucleotide modifications is one posttranscriptional mechanism of regulating RNA fate and function, and modifications including N6-methyladenosine (m6A), N1-methyladenosine (m1A), N5-methylcytidine (m5C), N4-acetylcytidine (ac4C), and pseudouridine (Ψ), dynamically regulate RNA stability and translation in diverse organisms. Little is known about RNA modifications and their machinery in Trypanosomatids, but we hypothesize that they regulate parasite gene expression and are vital for survival. Here, we identified Trypanosomatid homologs for writers of m1A, m5C, ac4C, and Ψ and analyze their evolutionary relationships. We systematically review the evidence for their functions and assess their potential use as therapeutic targets. This work provides new insights into the roles of these proteins in Trypanosomatid parasite biology and treatment of the diseases they cause and illustrates that Trypanosomatids provide an excellent model system to study RNA modifications, their molecular, cellular, and biological consequences, and their regulation and interplay.


[Oxygen concentrators: evolution of inspired concentration of oxygen and repercussions in an anesthetized patient with CO2 absorber system. Pilot study.].
Resolution 1355/92 of the Conselho Federal de Medicina approved minimal standards for the installation and operation of oxygen concentrators (PSA) and recommended University Hospitals to undertake a prospective analysis in order to improve the system. It motivated this pilot study whose objective was to determine the clinical viability of using PSA oxygen by analyzing the variation in oxygen concentration in the fresh gas flow (FGF) outlet and in the inspired concentration of oxygen. An observational study with 30 patients, ASA I, undergoing upper abdominal surgeries using a CO2 absorber system and fresh gas flow (FGF) O(2)93 at 500 mL.min-1. Weight, age, type and duration of the surgery, inspired and expired fraction of CO2 (FiCO2, P ET CO2); inspired fraction of O2 (FiO2); and O2 concentration (O2ent) in the FGF. The following parameters were measured after intubation and every 10 minutes until the end of the procedure: P ET CO2, FiO2, and O2. Results underwent statistical analysis and p < 0.05 was considered significant. The inspired fraction of carbon dioxide was equal to zero in all patients and moments of the study, but there was a significant reduction in P ET CO2 with time. The variables O2ent and FiO2 had similar tendencies with time (p = 0.1283), but the variable O2ent presented higher means (p < 0.001); evolution of mean O2ent and FiO2 was observed (p < 0.05). This study demonstrated that the use of PSA oxygen, within the conditions proposed for the experiment, is safe and induced a progressive increase in O(2)93 in the FGF and of FiO2.


Association between Subclinical Malaria Infection and Inflammatory Host Response in a Pre-Elimination Setting.
Subclinical infections in endemic areas of Southeast Asia sustain malaria transmission. These asymptomatic infections might sustain immunity against clinical malaria and have been considered benign for the host, but if they are associated with chronic low-grade inflammation this could be harmful. We conducted a case-control study to explore the association between subclinical malaria and C-reactive protein (CRP), an established biomarker of inflammation. Blood samples from asymptomatic villagers in Pailin, Western Cambodia were tested for malaria by high-volume ultra-sensitive polymerase chain reaction (uPCR) to determine the Plasmodium species. Plasma CRP concentration was measured in 328 individuals with parasitaemia (cases) and compared with: i) the same individual's value at the first time point when they had no detectable parasites (n = 282); and ii) age- sex- and village-matched controls (n = 328) free of Plasmodium infection. Plasma CRP concentrations were compared against thresholds of 3mg/L and 10mg/L. Subgroup analysis was carried out for cases with P vivax and P falciparum mono-infections. Median plasma CRP level for all samples was 0.59mg/L (interquartile range: 0.24-1.64mg/L). CRP concentrations were higher in parasitaemic individuals compared with same-person-controls (p = 0.050); and matched-controls (p = 0.025). 4.9% of samples had CRP concentrations above 10mg/L and 14.6% were above 3mg/L. Cases were more likely to have plasma CRP concentrations above these thresholds than age/sex matched controls, odds ratio 3.5 (95%CI 1.5-9.8) and 1.8 (95%CI 1.1-2.9), respectively. Amongst cases, parasite density and CRP were positively correlated (p<0.001), an association that remained significant when controlling for age and fever. Individuals with P.vivax mono-infections had the highest plasma CRP concentrations with the greatest association with parasitaemia. In this setting persistent malaria infections in asymptomatic individuals were associated with moderately elevated plasma CRP concentrations; chiefly evident in cases with P.vivax mono-infections. As well as interrupting malaria transmission within the community, treatment of asymptomatic malaria infections, in particular radical cure of vivax malaria, may benefit the health of infected individuals.


[DNA damage assessment and biological monitoring of occupational exposure to organic solvents, 2006].
Exposure to solvents is one of the highest potential risks for millions of workers in the world; they can generate substantial environmental pollution leading to outbreaks of public health problems. Blood levels were determined for metabolites of benzene, toluene and xylene, and polymorphisms for enzymes CYP2E1, GSTM1, GSTT1 were characterized. Damage to DNA was assessed by the comet assay for exposure to organic solvents. A cross-sectional study was undertaken in 90 employees from 5 different companies. A survey form was administered; blood was sampled to detect the genetic polymorphisms and to apply the comet assay (single cell gel electrophoresis) to detect DNA fragmentation. The concentrations of phenol, ortho and meta methylhippuric acids were measured in urine. Statistical analyses explored the possible associations. The percentage of workers directly exposed to solvents was 34.4%. In this group, the evidence indicated concentrations higher than the permitted limits: 3.3% for phenol, 6.6% for hippuric acid, 3.3% for ortho-methylhippuric acid and 36.7% for metamethylhippuric acid. In the Comet assay, the length of the comets tail was greater than average (19.5 ìm) in exposed subjects, and the percentage of cells with mild damage (19.0%) (p=0.0007) was higher. The percentage of exposed individuals with absent genotypes for enzymes GSTT1 and GSTM1 was 46.7% and 56.8% respectively. Exposure biomarkers have become fundamental tools for the evaluation of risk associated with exposure to toxic agents.


Managerial guidelines to support parents during the hospitalisation of their children in a private paediatric unit.
The purpose of this article is to describe managerial guidelines to support parents with the hospitalisation of their child in a private paediatric unit. The hospitalisation of a child is regarded as a major stressor for both parents and child. The role of the family in participating in a child's illness is slowly being recognised (Kibel & Wagstaff 2001:544), but the South African government per se has not yet issued any formal reports on parental participation in the hospitalisation process. The study explored and described the nursing care experiences of parents regarding the hospitalisation of their child in a paediatric unit; managerial guidelines to support parents with their lived experiences of their child's hospitalisation in a paediatric unit. To achieve the purpose and the objectives of the research, an interpretive-phenomenological qualitative approach was used in the research design and methods. Research was conducted through unstructured individual interviews, narrative diaries and field notes and data were analysed through open-coding (Tesch, 1990). Parents were asked to respond to the question "How did you experience your child's hospitalisation in the paediatric ward", followed by probing when the responses of the parents were ambiguous. Purposive sampling was used to achieve saturation of data and seven parents were interviewed and fifteen parents completed narrative diaries. The model of Lincoln and Guba (1985) was used to ensure trustworthiness. Ethical considerations were maintained throughout the study and consent was obtained from the respondents. The recommendations of the research were that attention should be given to 1) empowering parents to participate in their child's care; 2) guiding nursing personnel to plan the discharge process; 3) including parents in the unit routine; 4) fostering a trusting relationship with parents; 5) promoting the communication of information; and 6) creating a therapeutic environment for parents.


Modeling reciprocal effects in medical research: Critical discussion on the current practices and potential alternative models.
Longitudinal designs provide a strong inferential basis for uncovering reciprocal effects or causality between variables. For this analytic purpose, a cross-lagged panel model (CLPM) has been widely used in medical research, but the use of the CLPM has recently been criticized in methodological literature because parameter estimates in the CLPM conflate between-person and within-person processes. The aim of this study is to present some alternative models of the CLPM that can be used to examine reciprocal effects, and to illustrate potential consequences of ignoring the issue. A literature search, case studies, and simulation studies are used for this purpose. We examined more than 300 medical papers published since 2009 that applied cross-lagged longitudinal models, finding that in all studies only a single model (typically the CLPM) was performed and potential alternative models were not considered to test reciprocal effects. In 49% of the studies, only two time points were used, which makes it impossible to test alternative models. Case studies and simulation studies showed that the CLPM and alternative models often produce different (or even inconsistent) parameter estimates for reciprocal effects, suggesting that research that relies only on the CLPM may draw erroneous conclusions about the presence, predominance, and sign of reciprocal effects. Simulation studies also showed that alternative models are sometimes susceptible to improper solutions, even when reseachers do not misspecify the model.


Direct chemical vapor deposition of graphene on dielectric surfaces.
Direct deposition of graphene on various dielectric substrates is demonstrated using a single-step chemical vapor deposition process. Single-layer graphene is formed through surface catalytic decomposition of hydrocarbon precursors on thin copper films predeposited on dielectric substrates. The copper films dewet and evaporate during or immediately after graphene growth, resulting in graphene deposition directly on the bare dielectric substrates. Scanning Raman mapping and spectroscopy, scanning electron microscopy, and atomic force microscopy confirm the presence of continuous graphene layers on tens of micrometer square metal-free areas. The revealed growth mechanism opens new opportunities for deposition of higher quality graphene films on dielectric materials.


Serum vitamin D and risk of bladder cancer in the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening trial.
The one previous prospective study of vitamin D status and risk of urinary bladder cancer found that male smokers with low serum 25-hydroxy-vitamin D [25(OH)D] were at a nearly two-fold increased risk. We conducted an analysis of serum 25(OH)D and risk of bladder cancer in the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Study and examined whether serum vitamin D binding protein (DBP) concentration confounded or modified the association. Three hundred and seventy-five cases of bladder cancer were matched 1:1 with controls based on age (±5 years), race, sex, and date of blood collection (±30 days). Conditional logistic regression was used to estimate ORs and 95% confidence intervals (CI) of bladder cancer by prediagnosis levels of 25(OH)D. We found no strong or statistically significant association between serum 25(OH)D and bladder cancer risk (Q1 vs. Q4: OR, 0.84; 95% CI, 0.52-1.36; P(trend) = 0.56). Further adjustment for, or stratification by, serum DBP did not alter the findings, nor was there a main effect association between DBP and risk. In contrast to an earlier report, we observed no association between vitamin D status and risk of bladder cancer; this difference could be due to the inclusion of women and nonsmokers in the current study population or due to the differences in the distribution of vitamin D concentrations between the two study populations. These findings may contribute to future meta-analyses and help elucidate whether the vitamin D-bladder cancer association varies across populations.


Rat respiratory coronavirus infection: replication in airway and alveolar epithelial cells and the innate immune response.
The rat coronavirus sialodacryoadenitis virus (SDAV) causes respiratory infection and provides a system for investigating respiratory coronaviruses in a natural host. A viral suspension in the form of a microspray aerosol was delivered by intratracheal instillation into the distal lung of 6-8-week-old Fischer 344 rats. SDAV inoculation produced a 7 % body weight loss over a 5 day period that was followed by recovery over the next 7 days. SDAV caused focal lesions in the lung, which were most severe on day 4 post-inoculation (p.i.). Immunofluorescent staining showed that four cell types supported SDAV virus replication in the lower respiratory tract, namely Clara cells, ciliated cells in the bronchial airway and alveolar type I and type II cells in the lung parenchyma. In bronchial alveolar lavage fluid (BALF) a neutrophil influx increased the population of neutrophils to 45 % compared with 6 % of the cells in control samples on day 2 after mock inoculation. Virus infection induced an increase in surfactant protein SP-D levels in BALF of infected rats on days 4 and 8 p.i. that subsided by day 12. The concentrations of chemokines MCP-1, LIX and CINC-1 in BALF increased on day 4 p.i., but returned to control levels by day 8. Intratracheal instillation of rats with SDAV coronavirus caused an acute, self-limited infection that is a useful model for studying the early events of the innate immune response to respiratory coronavirus infections in lungs of the natural virus host.


Effect of Gold Nanoparticle Size on Their Properties as Contrast Agents for Computed Tomography.
Computed tomography (CT) is one of the most commonly used clinical imaging modalities. There have recently been many reports of novel contrast agents for CT imaging. In particular, the development of gold nanoparticles (AuNP) as CT contrast agents is a topic of intense interest. AuNP have favorable characteristics for this application such as high payloads of contrast generating material, strong X-ray attenuation, excellent biocompatibility, tailorable surface chemistry, and tunable sizes and shapes. However, there have been conflicting reports on the role of AuNP size on their contrast generation for CT. We therefore sought to extensively investigate the AuNP size-CT contrast relationship. In order to do this, we synthesized AuNP with sizes ranging from 4 to 152 nm and capped them with 5 kDa m-PEG. The contrast generation of AuNP of different sizes was investigated with three clinical CT, a spectral photon counting CT (SPCCT) and two micro CT systems. X-ray attenuation was quantified as attenuation rate in Hounsfield units per unit concentration (HU/mM). No statistically significant difference in CT contrast generation was found among different AuNP sizes via phantom imaging with any of the systems tested. Furthermore, in vivo imaging was performed in mice to provide insight into the effect of AuNP size on animal biodistribution at CT dose levels, which has not previously been explored. Both in vivo imaging and ex vivo analysis with inductively coupled plasma optical emission spectroscopy (ICP-OES) indicated that AuNP that are 15 nm or smaller have long blood circulation times, while larger AuNP accumulated in the liver and spleen more rapidly. Therefore, while we observed no AuNP size effect on CT contrast generation, there is a significant effect of size on AuNP diagnostic utility.


Colonisation with methicillin-resistant Staphylococcus aureus and associated factors among nurses with occupational skin diseases.
To evaluate the prevalence of colonisation with methicillin-resistant Staphylococcus aureus (MRSA), associated factors and the effectiveness of decolonisation procedures among nurses with occupational skin diseases (OSD). In a retrospective cohort study, the medical records of 319 nurses from Germany who were screened consecutively for MRSA when participating in a tertiary individual prevention programme (TIP) for severe OSD between July 2009 and December 2014 were evaluated. 90.3% of nurses with severe OSD suffered from hand eczema. 43 were colonised with MRSA on admission (13.5%), mainly in the nose (n=35, 81.4%). However, the hands were affected in more than half of the MRSA carriers (n=24, 55.8%). Risk factors for MRSA colonisation were atopic skin diathesis (OR 2.01, 95% CI 1.03 to 3.92, p=0.049) and presence of atopic dermatitis on other body parts than the hands (OR 4.33, 95% CI 2.23 to 8.43, p<0.001). Hand eczema was significantly more severe in MRSA carriers than in non-carriers (OR 1.23, 95% CI 1.10 to 1.37, p<0.001) and showed a higher prevalence of vesicles, erosions or fissures. MRSA eradication was successful in 67.4% after the first attempt. Nurses with OSD have a twofold to threefold higher prevalence of MRSA colonisation than what has been reported for point-prevalence screenings among healthcare workers in Germany. Atopic skin diathesis, atopic dermatitis and severe hand eczema are the main risk factors. Thus, prevention and treatment of OSD could be important elements in reduction of colonisation with MRSA among nurses and transmission to others.


Comparative effectiveness of sulfonylurea and metformin monotherapy on cardiovascular events in type 2 diabetes mellitus: a cohort study.
The effects of sulfonylureas and metformin on outcomes of cardiovascular disease (CVD) in type 2 diabetes are not well-characterized. To compare the effects of sulfonylureas and metformin on CVD outcomes (acute myocardial infarction and stroke) or death. Retrospective cohort study. National Veterans Health Administration databases linked to Medicare files. Veterans who initiated metformin or sulfonylurea therapy for diabetes. Patients with chronic kidney disease or serious medical illness were excluded. Composite outcome of hospitalization for acute myocardial infarction or stroke, or death, adjusted for baseline demographic characteristics; medications; cholesterol, hemoglobin A1c, and serum creatinine levels; blood pressure; body mass index; health care utilization; and comorbid conditions. Among 253 690 patients initiating treatment (98 665 with sulfonylurea therapy and 155 025 with metformin therapy), crude rates of the composite outcome were 18.2 per 1000 person-years in sulfonylurea users and 10.4 per 1000 person-years in metformin users (adjusted incidence rate difference, 2.2 [95% CI, 1.4 to 3.0] more CVD events with sulfonylureas per 1000 person-years; adjusted hazard ratio [aHR], 1.21 [CI, 1.13 to 1.30]). Results were consistent for both glyburide (aHR, 1.26 [CI, 1.16 to 1.37]) and glipizide (aHR, 1.15 [CI, 1.06 to 1.26]) in subgroups by CVD history, age, body mass index, and albuminuria; in a propensity score-matched cohort analysis; and in sensitivity analyses. Most of the veterans in the study population were white men; data on women and minority groups were limited but reflective of the Veterans Health Administration population. Use of sulfonylureas compared with metformin for initial treatment of diabetes was associated with an increased hazard of CVD events or death. Agency for Healthcare Research and Quality and the U.S. Department of Health and Human Services.


Phenotypic switching of in vitro mandibular condylar cartilage during matrix mineralization.
In order to analyze the phenotypic conversion of chondrocytes, mandibular condyles of mice and rabbits were cultured under cell and organ culture systems, and then examined by a combination of morphological and biochemical procedures. In organ culture, mandibular condylar cartilage (MCC) obtained from newborn mice began to mineralize from the central zone and then progressively widened towards the peripheral zone. Electron microscopic observations showed that with the increasing duration of the organ culture, chondrocytes at the central zone converted into spindle-shaped osteoblastic cells accompanying the formation of the bone type of thick-banded collagen fibrils. To obtain a better understanding of the chondrocytic conversion, immunolocalizations for type I and type X collagens and osteocalcin (OC) were examined in mouse MCC cells in cell culture. Type X collagen and OC were expressed almost simultaneously at the late stage of culture, and type I collagen was detected along the calcified nodules after the production of these proteins. Northern blot analysis in cell cultures of rabbit MCC indicated that type II collagen and alkaline phosphatase (ALPase) messenger ribonucleic acids (mRNAs) were highly expressed at day 7, but subsequently decreased. In contrast, mRNA for type I collagen was expressed at a low level on day 7 and peaked on day 12. The present results suggest that, morphologically and biochemically, cellular modification in MCC cells under culture conditions occurs at a cellular morphological level and also at marker-gene-expression level.


Constructing a new approach to developing evidence-based practice with nurses and older people.
Providing evidence-based nursing care to older people is central to the international development agenda. This paper is a report on the first 5 years (2000-2005) of a participatory research project, the purpose of which was to collaborate with practitioners and older people to develop approaches to promote the attainment of evidence-based nursing care across Scotland. Many theoretical influences shaped the design of this action research study including realistic evaluation, participatory social learning theory, and descriptions of communities of practice. Multiple methods of data collection were used during four action cycles. The inaugural community of practice comprised 30 nurses, a second group of 30 nurses joined midway, followed by a third group of 15 nurses, and finally, an older person-carer community of 21 members was established. Project outputs included the construction of an internet-based, practice-development college. A procedural model for developing and demonstrating care guidance drawn from a diversity of evidence and reflective of an agreed set of principles was piloted and endorsed by the national standard setting agency. A preliminary version of a promising approach to practice development, "the Caledonian Model," was delineated for future testing and refinement. This work indicates the merits of using participatory research to find solutions to the challenge of promoting evidence-based practice. Evaluation data suggest that in combination, the approaches developed in this project empower nurses to work with older people to champion developments even in seemingly unfavorable conditions.


Economical synthesis of 13C-labeled opiates, cocaine derivatives and selected urinary metabolites by derivatization of the natural products.
The illegal use of opiates and cocaine is a challenge world-wide, but some derivatives are also valuable pharmaceuticals. Reference samples of the active ingredients and their metabolites are needed both for controlling administration in the clinic and to detect drugs of abuse. Especially, (13)C-labeled compounds are useful for identification and quantification purposes by mass spectroscopic techniques, potentially increasing accuracy by minimizing ion alteration/suppression effects. Thus, the synthesis of [acetyl-(13)C4]heroin, [acetyl-(13)C4-methyl-(13)C]heroin, [acetyl-(13)C2-methyl-(13)C]6-acetylmorphine, [N-methyl-(13)C-O-metyl-(13)C]codeine and phenyl-(13)C6-labeled derivatives of cocaine, benzoylecgonine, norcocaine and cocaethylene was undertaken to provide such reference materials. The synthetic work has focused on identifying (13)C atom-efficient routes towards these derivatives. Therefore, the (13)C-labeled opiates and cocaine derivatives were made from the corresponding natural products.


Habitat Selection of Wintering Birds in Farm Ponds in Taoyuan, Taiwan.
Farm ponds or irrigation ponds, providing a vital habitat for diverse bird communities, are an environmental feature with characteristics that cross over typical urban and natural conditions. In this study, the species richness and community structure of irrigation ponds were characterized on the local and landscape scales. Within a landscape complex in the Taoyuan Tableland of Taiwan, 45 ponds were surveyed, ranging in areas from 0.2 to 20.47 ha. In total, 94 species and 15,053 individual birds were identified after surveying four times. The association between ponds and birds was determined to establish the effect of pond dimensions on species richness and community composition in the complex by comparing the responses of functional groups to pond configurations. Seven avian functional groups were identified. Compared with landbirds (i.e., families Alcedinidae, Apodidae, Icteridae, and Sturnidae), waterbirds (i.e., families Anatidae, Ardeidae, Charadriidae, Podicipedidae, and Scolopacidae) exhibited a stronger correlation with pond variables. Our study provides substantial evidence that these artificial ponds have influenced wintering waterbirds. The final results of this study may help stakeholders and land managers identify areas not to establish large-scale solar facilities considering waterbird habitats in pond areas.


Antibody-coupled monolithic silica microtips for highthroughput molecular profiling of circulating exosomes.
Exosome-mediated signal transportation plays a variety of critical roles in cancer progression and metastasis. From the aspect of cancer diagnosis, circulating exosomes are ideal resources of biomarkers because molecular features of tumor cells are transcribed on them. However, isolating pure exosomes from body fluids is time-consuming and still major challenge to be addressed for comprehensive profiling of exosomal proteins and miRNAs. Here we constructed anti-CD9 antibody-coupled highly porous monolithic silica microtips which allowed automated rapid and reproducible exosome extraction from multiple clinical samples. We applied these tips to explore lung cancer biomarker proteins on exosomes by analyzing 46 serum samples. The mass spectrometric quantification of 1,369 exosomal proteins identified CD91 as a lung adenocarcinoma specific antigen on exosomes, which was further validated with CD9-CD91 exosome sandwich ELISA measuring 212 samples. Our simple device can promote not only biomarker discovery studies but also wide range of omics researches about exosomes.


Teenagers response to threat of war and terror: gender and the role of social systems.
This study focuses on the role of gender and the way social systems (family and friends' support, stress attributed to parents, friends and others in the community) are perceived in contributing to the reaction to stressors of war and threat of terror among teenagers living in Israel along the Lebanese border. The study was implemented shortly after Israel's withdrawal from Lebanon with a sample of 353 teenagers. The study is based on the teenagers' self-report. The results reveals the following: (a) gender relates significantly to level of stress but not to general life satisfaction; (b) stress attributed to the parents and to the peers has a greater contribution to the level of stress than the supportive family atmosphere and peer support; (c) stress attributed to the parents and peers was found to be a mediator between gender and level of stress; (d) supportive family atmosphere was found to be a mediator between gender and one measurement of stress, while peer support was not found to have any mediating role. Special attention is given to the finding that stress attributed to parents and friends has greater significance than the support provided by them.


Effects of 1,4-phenylenebis(methylene)selenocyanate on mutagenesis and p53 protein expression in the tongue of lacI rats treated with 4-nitroquinoline-N-oxide.
Previously we showed that the organoselenium compound, 1,4-phenylenebis(methylene)selenocyanate (p-XSC)(1) inhibits 4-nitroquinoline-N-oxide (4-NQO)-induced tongue tumorigenesis in Fisher rats. Here we investigate possible mechanisms of this inhibition by monitoring mutagenesis and p53 protein levels in lacI and conventional Fisher rats treated with: (1) a carcinogenic dose of 4-NQO for 10 weeks in drinking water, (2) 4-NQO+p-XSC (15 ppm as selenium), and (3) 4-NQO followed by p-XSC. For mutagenesis studies, rats were euthanized at 7, 12 or 23 weeks after the start of 4-NQO. For studies on p53 levels, rats were euthanized at 11, 15 and 23 weeks. Appropriate controls were also monitored. In the 4-NQO-alone groups, the mutant fraction (MF) in the cII gene in tongue increased at least 50x background level. The MF (in units of mutants/10(5) plaque forming units) for the 7, 12, and 23 weeks 4-NQO groups were respectively, 184 +/- 88, 237 +/- 105, and 329 +/- 110. Thus, mutagenesis increased with length of exposure and post-treatment time. p-XSC modestly (ca. 15-30%) inhibited mutagenesis under all conditions. The inhibition reached significance at the last time point. When p-XSC was administered after 4-NQO, the MF was also modestly reduced. In 4-NQO-alone animals, levels of p53 in tongue (determined by Western blotting) were 1, 1.5 and 2.4 control levels at 10, 15 and 23 weeks, respectively. In the p-XSC+4-NQO group, the enhancement in p53 levels by 4-NQO treatment was decreased about 90% at 15 weeks and 45% (P<0.05) at 23 weeks, and by slightly smaller percentages in corresponding post-treatment groups. p-XSC alone did not alter p53 levels. As p53 levels generally increase in response to DNA damage, these results suggest that p-XSC reduces 4-NQO-induced DNA damage, resulting in reduced 4-NQO-induced mutagenesis and carcinogenesis. However, the fact that p-XSC is also effective when administered after 4-NQO, suggests additional mechanisms of inhibition exist.


Contradictory reasoning network: an EEG and FMRI study.
Contradiction is a cornerstone of human rationality, essential for everyday life and communication. We investigated electroencephalographic (EEG) and functional magnetic resonance imaging (fMRI) in separate recording sessions during contradictory judgments, using a logical structure based on categorical propositions of the Aristotelian Square of Opposition (ASoO). The use of ASoO propositions, while controlling for potential linguistic or semantic confounds, enabled us to observe the spatial temporal unfolding of this contradictory reasoning. The processing started with the inversion of the logical operators corresponding to right middle frontal gyrus (rMFG-BA11) activation, followed by identification of contradictory statement associated with in the right inferior frontal gyrus (rIFG-BA47) activation. Right medial frontal gyrus (rMeFG, BA10) and anterior cingulate cortex (ACC, BA32) contributed to the later stages of process. We observed a correlation between the delayed latency of rBA11 response and the reaction time delay during inductive vs. deductive reasoning. This supports the notion that rBA11 is crucial for manipulating the logical operators. Slower processing time and stronger brain responses for inductive logic suggested that examples are easier to process than general principles and are more likely to simplify communication.


Polymorphism identification and improved genome annotation of Brassica rapa through Deep RNA sequencing.
The mapping and functional analysis of quantitative traits in Brassica rapa can be greatly improved with the availability of physically positioned, gene-based genetic markers and accurate genome annotation. In this study, deep transcriptome RNA sequencing (RNA-Seq) of Brassica rapa was undertaken with two objectives: SNP detection and improved transcriptome annotation. We performed SNP detection on two varieties that are parents of a mapping population to aid in development of a marker system for this population and subsequent development of high-resolution genetic map. An improved Brassica rapa transcriptome was constructed to detect novel transcripts and to improve the current genome annotation. This is useful for accurate mRNA abundance and detection of expression QTL (eQTLs) in mapping populations. Deep RNA-Seq of two Brassica rapa genotypes-R500 (var. trilocularis, Yellow Sarson) and IMB211 (a rapid cycling variety)-using eight different tissues (root, internode, leaf, petiole, apical meristem, floral meristem, silique, and seedling) grown across three different environments (growth chamber, greenhouse and field) and under two different treatments (simulated sun and simulated shade) generated 2.3 billion high-quality Illumina reads. A total of 330,995 SNPs were identified in transcribed regions between the two genotypes with an average frequency of one SNP in every 200 bases. The deep RNA-Seq reassembled Brassica rapa transcriptome identified 44,239 protein-coding genes. Compared with current gene models of B. rapa, we detected 3537 novel transcripts, 23,754 gene models had structural modifications, and 3655 annotated proteins changed. Gaps in the current genome assembly of B. rapa are highlighted by our identification of 780 unmapped transcripts. All the SNPs, annotations, and predicted transcripts can be viewed at http://phytonetworks.ucdavis.edu/.


Oxidative damage in mitochondrial DNA is not extensive.
Since 1988 several research groups have reported greater levels of oxidative damage in mitochondrial DNA than in nuclear DNA, while others have suggested that the greater damage in mtDNA might be due to artifactual oxidation. The popular theory that mtDNA is more heavily damaged in vivo than nDNA does not stand on firm ground. Using an improved GC-MS method and pure mtDNA, our analyses revealed that the damage level in mtDNA is not higher, and may be somewhat lower, than that in nDNA.


Maternal neglect results in reduced telomerase activity and increased oxidative load in rats.
A growing number of studies in humans have linked chronic stress, particularly during early life, to telomere shortening and increased oxidative stress. The effect of stress on telomerase activity, however, is understudied. Given the importance of telomere attrition in a wide range of diseases and immunosenescence, further research to elucidate the mechanisms by which stress alters telomere dynamics is required. However, animal studies are lacking, and it is not clear whether widely used stress models reliably mimic the accelerated telomere shortening observed humans. To this end, we evaluated the effect of maternal separation with early weaning (MSEW) on telomere length, telomerase activity, and oxidative load in rats. A total of 45 animals were used, (17 control: 3 males and 11 females and 28 MSEW: 11 males, 17 females), which were then sacrificed one year after birth. Importantly, we determined that telomerase activity measured in plasma was significantly decreased in the MSEW group, along with a non-significant reduction in telomere length from whole blood cells. We also examined the levels of three oxidative markers: plasma malondialdehyde, glutathione in erythrocytes, and plasma catalase activity. Malondialdehyde was found to be elevated in the plasma, indicating increased lipid peroxidation. Interestingly, while the antioxidant glutathione was upregulated, catalase activity remained unchanged. Our findings indicate that the rat MSEW model induces chronic changes to telomere dynamics and oxidative load and can capitulate long term aspects of human childhood stress.


A therapeutic effect of cbd-enriched ointment in inflammatory skin diseases and cutaneous scars.
To investigate the therapeutic effect of CBD-ointment administered on severe skin chronic diseases and/or on their outcome scars. A spontaneous, anecdotal, retrospective study of 20 patients with two most frequent skin disorders: psoriasis (n: 5 patients), atopic dermatitis (n: 5) and resulting outcome scars (n: 10). The subjects were instructed to administer topical CBD-enriched ointment to lesioned skin areas twice daily for three months treatment. Based on skin evaluations (hydration, TEWL, elasticity), clinical questionnaires (SCORAD, ADI, PASI), and supported by photographic data and investigators' clinical assessment, the results showed that topical treatment with CBD-enriched ointment significantly improved the skin parameters, the symptoms and also the PASI index score. No irritant or allergic reactions were documented during the period treatment. The topical administration of CBD ointment, without any THC, is a safe and effective non-invasive alternative for improve the quality of life in patients with some skin disorders, especially on inflammatory background.


Long-term change in a behavioural trait: truncated spawning distribution and demography in Northeast Arctic cod.
Harvesting may be a potent driver of demographic change and contemporary evolution, which both may have great impacts on animal populations. Research has focused on changes in phenotypic traits that are easily quantifiable and for which time series exist, such as size, age, sex, or gonad size, whereas potential changes in behavioural traits have been under-studied. Here, we analyse potential drivers of long-term changes in a behavioural trait for the Northeast Arctic stock of Atlantic cod Gadus morhua, namely choice of spawning location. For 104 years (1866-1969), commercial catches were recorded annually and reported by county along the Norwegian coast. During this time period, spawning ground distribution has fluctuated with a trend towards more northerly spawning. Spawning location is analysed against a suite of explanatory factors including climate, fishing pressure, density dependence, and demography. We find that demography (age or age at maturation) had the highest explanatory power for variation in spawning location, while climate had a limited effect below statistical significance. As to potential mechanisms, some effects of climate may act through demography, and explanatory variables for demography may also have absorbed direct evolutionary change in migration distance for which proxies were unavailable. Despite these caveats, we argue that fishing mortality, either through demographic or evolutionary change, has served as an effective driver for changing spawning locations in cod, and that additional explanatory factors related to climate add no significant information.


SIRT1 enhances glucose tolerance by potentiating brown adipose tissue function.
SIRT1 has been proposed to be a key signaling node linking changes in energy metabolism to transcriptional adaptations. Although SIRT1 overexpression is protective against diverse metabolic complications, especially in response to high-fat diets, studies aiming to understand the etiology of such benefits are scarce. Here, we aimed to identify the key tissues and mechanisms implicated in the beneficial effects of SIRT1 on glucose homeostasis. We have used a mouse model of moderate SIRT1 overexpression, under the control of its natural promoter, to evaluate glucose homeostasis and thoroughly characterize how different tissues could influence insulin sensitivity. Mice with moderate overexpression of SIRT1 exhibit better glucose tolerance and insulin sensitivity even on a low fat diet. Euglycemic-hyperinsulinemic clamps and in-depth tissue analyses revealed that enhanced insulin sensitivity was achieved through a higher brown adipose tissue activity and was fully reversed by housing the mice at thermoneutrality. SIRT1 did not influence brown adipocyte differentiation, but dramatically enhanced the metabolic transcriptional responses to β3-adrenergic stimuli in differentiated adipocytes. Our work demonstrates that SIRT1 improves glucose homeostasis by enhancing BAT function. This is not consequent to an alteration in the brown adipocyte differentiation process, but as a result of potentiating the response to β3-adrenergic stimuli.


Just add water: cannabinoid discrimination in a water T-maze with FAAH(-/-) and FAAH(+/+) mice.
Incomplete overlap in the discriminative stimulus effects of Δ-tetrahydrocannabinol (THC) and the endocannabinoids, anandamide (AEA) and 2-arachidonoylglycerol has been reported in food-reinforced tasks. The aim of this study was to examine cannabinoid discriminative stimulus effects in a nonappetitive procedure. Adult male mice lacking the gene for AEA's major metabolic enzyme, fatty acid amide hydrolase (FAAH), and FAAH mice were trained to discriminate THC or AEA in a water T-maze, in which the response was swimming to an escape platform on the injection-appropriate side. JZL184, a monoacylglycerol lipase inhibitor, was also tested. FAAH mice showed faster acquisition than FAAH mice. THC and AEA fully substituted, with only minor cross-procedure potency variations. Incomplete substitution of JZL184 was observed in THC-trained FAAH mice in the water-maze task, as contrasted with full substitution in a food-reinforced nose-poke procedure. Stress-induced changes in AEA and/or 2-arachidonoylglycerol concentrations in the brain may have mediated this attenuation. JZL184 also partially substituted in AEA-trained FAAH mice in the water maze, suggesting incomplete overlap in the stimulus effects of AEA and JZL184. Through the use of a novel water-maze procedure, the present study supports the work of previous behavioral pharmacologists in showing the robustness of the discrimination paradigm.


Prions: pathogenesis and reverse genetics.
Spongiform encephalopathies are a group of infectious neurodegenerative diseases. The infectious agent that causes transmissible spongiform encephalopathies was termed prion by Stanley Prusiner. The prion hypothesis states that the partially protease-resistant and detergent-insoluble prion protein (PrPsc) is identical with the infectious agent, and lacks any detectable nucleic acids. Since the latter discovery, transgenic mice have contributed many important insights into the field of prion biology. The prion protein (PrPc) is encoded by the Prnp gene, and disruption of Prnp leads to resistance to infection by prions. Introduction of mutant PrPc genes into PrPc-deficient mice was used to investigate structure-activity relationships of the PrPc gene with regard to scrapie susceptibility. Ectopic expression of PrPc in PrPc knockout mice proved a useful tool for the identification of host cells competent for prion replication. Finally, the availability of PrPc knockout and transgenic mice overexpressing PrPc allowed selective reconstitution experiments aimed at expressing PrPc in neurografts or in specific populations of hemato- and lymphopoietic cells. The latter studies helped in elucidating some of the mechanisms of prion spread and disease pathogenesis.