Butter increased total and LDL cholesterol compared with olive oil but resulted in higher HDL cholesterol compared with a habitual diet.
Butter is known to have a cholesterol-raising effect and, therefore, has often been included as a negative control in dietary studies, whereas the effect of moderate butter intake has not been elucidated to our knowledge. We compared the effects of moderate butter intake, moderate olive oil intake, and a habitual diet on blood lipids, high-sensitivity C-reactive protein (hsCRP), glucose, and insulin. The study was a controlled, double-blinded, randomized 2 × 5-wk crossover dietary intervention study with a 14-d run-in period during which subjects consumed their habitual diets. The study included 47 healthy men and women (mean ± SD total cholesterol: 5.22 ± 0.90 mmol/L) who substituted a part of their habitual diets with 4.5% of energy from butter or refined olive oil. Study subjects were 70% women with a mean age and body mass index (in kg/m²) of 40.4 y and 23.5, respectively. Butter intake increased total cholesterol and LDL cholesterol more than did olive oil intake (P < 0.05) and the run-in period (P < 0.005 and P < 0.05, respectively) and increased HDL cholesterol compared with the run-in period (P < 0.05). No difference in effects was observed for triacylglycerol, hsCRP, insulin, and glucose concentrations. The intake of saturated fatty acids was significantly higher in the butter period than in the olive oil and run-in periods (P < 0.0001). Moderate intake of butter resulted in increases in total cholesterol and LDL cholesterol compared with the effects of olive oil intake and a habitual diet (run-in period). Furthermore, moderate butter intake was also followed by an increase in HDL cholesterol compared with the habitual diet. We conclude that hypercholesterolemic people should keep their consumption of butter to a minimum, whereas moderate butter intake may be considered part of the diet in the normocholesterolemic population.


Neurotrypsin cleaves agrin locally at the synapse.
The synaptic serine protease neurotrypsin is considered to be essential for the establishment and maintenance of cognitive brain functions, because humans lacking functional neurotrypsin suffer from severe mental retardation. Neurotrypsin cleaves agrin at two homologous sites, liberating a 90-kDa and a C-terminal 22-kDa fragment from the N-terminal moiety of agrin. Morphological analyses indicate that neurotrypsin is contained in presynaptic terminals and externalized in association with synaptic activity, while agrin is localized to the extracellular space at or in the vicinity of the synapse. Here, we present a detailed biochemical analysis of neurotrypsin-mediated agrin cleavage in the murine brain. In brain homogenates, we found that neurotrypsin exclusively cleaves glycanated variants of agrin. Studies with isolated synaptosomes obtained by subcellular fractionation from brains of wild-type and neurotrypsin-overexpressing mice revealed that neurotrypsin-dependent cleavage of agrin was concentrated at synapses, where the most heavily glycanated variants of agrin predominate. Because agrin has been shown to play an important role in the formation and the maintenance of excitatory synapses in the central nervous system, its local cleavage at the synapse implicates the neurotrypsin/agrin system in the regulation of adaptive reorganizations of the synaptic circuitry in the context of cognitive functions, such as learning and memory.


Possible role of natural selection in the formation of tandem-repetitive noncoding DNA.
A simulation model of sequence-dependent amplification, unequal crossing over and mutation is analyzed. This model predicts the spontaneous formation of tandem-repetitive patterns of noncoding DNA from arbitrary sequences for a wide range of parameter values. Natural selection is found to play an essential role in this self-organizing process. Natural selection which is modeled as a mechanism for controlling the length of a nucleotide string but not the sequence itself favors the formation of tandem-repetitive structures. Two measures of sequence heterogeneity, inter-repeat variability and repeat length, are analyzed in detail. For fixed mutation rate, both inter-repeat variability and repeat length are found to increase with decreasing rates of (unequal) crossing over. The results are compared with data on micro-, mini- and satellite DNAs. The properties of minisatellites and satellite DNAs resemble the simulated structures very closely. This suggests that unequal crossing over is a dominant long-range ordering force which keeps these arrays homogeneous even in regions of very low recombination rates, such as at satellite DNA loci. Our analysis also indicates that in regions of low rates of (unequal) crossing over, inter-repeat variability is maintained at a low level at the expense of much larger repeat units (multimeric repeats), which are characteristic of satellite DNA. In contrast, the microsatellite data do not fit the proposed model well, suggesting that unequal crossing over does not act on these very short tandem arrays.


Exclusion processes with internal states.
We introduce driven exclusion processes with internal states that serve as generic transport models in various contexts, ranging from molecular or vehicular traffic on parallel lanes to spintronics. The ensuing nonequilibrium steady states are controllable by boundary as well as bulk rates. A striking polarization phenomenon accompanied by domain wall motion and delocalization is discovered within a mesoscopic scaling. We quantify this observation within an analytic description providing exact phase diagrams. Our results are confirmed by stochastic simulations.


Three questions that patients can ask to improve the quality of information physicians give about treatment options: a cross-over trial.
To test the effect of three questions (what are my options? what are the benefits and harms? and how likely are these?), on information provided by physicians about treatment options. We used a cross-over trial using two unannounced standardized patients (SPs) simulating a presentation of mild-moderate depression. One SP was assigned the intervention role (asking the questions), the other the control role. An intervention and control SP visited each physician, order allocated randomly. The study was conducted in family practices in Sydney, Australia, during 2008-09. Data were obtained from consultation audio-recordings. Information about treatment options and patient involvement were analyzed using the Assessing Communication about Evidence and Patient Preferences (ACEPP) tool and the OPTION tool. Thirty-six SP visits were completed (18 intervention, 18 control). Scores were higher in intervention consultations than controls: ACEPP scores 21.4 vs. 16.6, p<0.001, difference 4.7 (95% CI 2.3-7.0) and OPTION scores 36 vs. 25, p=0.001, difference 11.5 (95% CI 5.1-17.8), indicating greater information provision and behavior supporting patient involvement. Asking these three questions improved information given by family physicians and increased physician facilitation of patient involvement. Practice implications. These questions can drive evidence-based practice, strengthen patient-physician communication, and improve safety and quality.


Comparison of Cardiovascular Risk Factors among Somalis Living in Norway and Somaliland.
We aimed to assess and compare cardiovascular disease (CVD) risk factors and predict the future risk of CVD among Somalis living in Norway and Somaliland. We included participants (20-69 years) from two cross-sectional studies among Somalis living in Oslo (n = 212) and Hargeisa (n = 1098). Demographic data, history of CVD, smoking, alcohol consumption, anthropometric measures, blood pressure, fasting serum glucose, and lipid profiles were collected. The predicted 10-year risk of CVD was calculated using Framingham risk score models. In women, systolic and diastolic blood pressure were significantly higher in Hargeisa compared to Oslo (p < 0.001), whereas no significant differences were seen in men. The ratio of total cholesterol to high-density lipoprotein (HDL) cholesterol was significantly higher in Hargeisa compared to Oslo among both men (4.4 versus 3.9, p = 0.001) and women (4.1 versus 3.3, p < 0.001). Compared to women, men had higher Framingham risk scores, but there were no significant differences in Framingham risk scores between Somalis in Oslo and Hargeisa. In spite of the high body mass index (BMI) in Oslo, most CVD risk factors were higher among Somali women living in Hargeisa compared to those in Oslo, with similar patterns suggested in men. However, the predicted CVD risks based on Framingham models were not different between the locations.


MR neurography in ulnar neuropathy as surrogate parameter for the presence of disseminated neuropathy.
Patients with ulnar neuropathy of unclear etiology occasionally present with lesion extension from elbow to upper arm level on MRI. This study investigated whether MRI thereby distinguishes multifocal neuropathy from focal-compressive neuropathy at the elbow. This prospective study was approved by the institutional ethics committee and written informed consent was obtained from all participants. 122 patients with ulnar mononeuropathy of undetermined localization and etiology by clinical and electrophysiological examination were assessed by MRI at upper arm and elbow level using T2-weighted fat-saturated sequences at 3T. Twenty-one patients were identified with proximal ulnar nerve lesions and evaluated for findings suggestive of disseminated neuropathy (i) subclinical lesions in other nerves, (ii) unfavorable outcome after previous decompressive elbow surgery, and (iii) subsequent diagnosis of inflammatory or other disseminated neuropathy. Two groups served as controls for quantitative analysis of nerve-to-muscle signal intensity ratios: 20 subjects with typical focal ulnar neuropathy at the elbow and 20 healthy subjects. In the group of 21 patients with proximal ulnar nerve lesion extension, T2-w ulnar nerve signal was significantly (p<0.001) higher at upper arm level than in both control groups. A cut-off value of 1.92 for maximum nerve-to-muscle signal intensity ratio was found to be sensitive (86%) and specific (100%) to discriminate this group. Ten patients (48%) exhibited additional T2-w lesions in the median and/or radial nerve. Another ten (48%) had previously undergone elbow surgery without satisfying outcome. Clinical follow-up was available in 15 (71%) and revealed definitive diagnoses of multifocal neuropathy of various etiologies in four patients. In another eight, diagnoses could not yet be considered definitive but were consistent with multifocal neuropathy. Proximal ulnar nerve T2 lesions at upper arm level are detected by MRI and indicate the presence of a non-focal disseminated neuropathy instead of a focal compressive neuropathy.


The Pleural Effusion And Symptom Evaluation (PLEASE) study of breathlessness in patients with a symptomatic pleural effusion.
Pathophysiology changes associated with pleural effusion, its drainage and factors governing symptom response are poorly understood. Our objective was to determine: 1) the effect of pleural effusion (and its drainage) on cardiorespiratory, functional and diaphragmatic parameters; and 2) the proportion as well as characteristics of patients with breathlessness relief post-drainage. Prospectively enrolled patients with symptomatic pleural effusions were assessed at both pre-therapeutic drainage and at 24-36 h post-therapeutic drainage. 145 participants completed pre-drainage and post-drainage tests; 93% had effusions ≥25% of hemithorax. The median volume drained was 1.68 L. Breathlessness scores improved post-drainage (mean visual analogue scale (VAS) score by 28.0±24 mm; dyspnoea-12 (D12) score by 10.5±8.8; resting Borg score before 6-min walk test (6-MWT) by 0.6±1.7; all p<0.0001). The 6-min walk distance (6-MWD) increased by 29.7±73.5 m, p<0.0001. Improvements in vital signs and spirometry were modest (forced expiratory volume in 1 s (FEV1) by 0.22 L, 95% CI 0.18-0.27; forced vital capacity (FVC) by 0.30 L, 95% CI 0.24-0.37). The ipsilateral hemi-diaphragm was flattened/everted in 50% of participants pre-drainage and 48% of participants exhibited paradoxical or no diaphragmatic movement. Post-drainage, hemi-diaphragm shape and movement were normal in 94% and 73% of participants, respectively. Drainage provided meaningful breathlessness relief (VAS score improved ≥14 mm) in 73% of participants irrespective of whether the lung expanded (mean difference 0.14, 95% CI 10.02-0.29; p=0.13). Multivariate analyses found that breathlessness relief was associated with significant breathlessness pre-drainage (odds ratio (OR) 5.83 per standard deviation (sd) decrease), baseline abnormal/paralyzed/paradoxical diaphragm movement (OR 4.37), benign aetiology (OR 3.39), higher pleural pH (OR per sd increase 1.92) and higher serum albumin level (OR per sd increase 1.73). Breathlessness and exercise tolerance improved in most patients with only a small mean improvement in spirometry and no change in oxygenation. Breathlessness improvement was similar in participants with and without trapped lung. Abnormal hemi-diaphragm shape and movement were independently associated with relief of breathlessness post-drainage.


Dental injuries in association with facial fractures.
The aim of the study was to investigate the association between dental injuries and facial fractures. We performed a prospective study of 273 patients examined at a level 1 trauma center in Switzerland from September 2005 until August 2006 who had facial fractures. Medical history and clinical and radiologic examination findings were recorded to evaluate demographics, etiology, presentation, and type of facial fracture, as well as its relationship to dental injury site and type. In 273 patients with dentition, we recorded 339 different facial fractures. Of these patients, 130 (47.5%) sustained a fracture in the non-tooth-bearing region, 44 (16%) had a fractured maxilla, and 65 (24%) had a fractured mandible. Among 224 patients with dentition who had a facial fracture in only 1 compartment, 140 injured teeth were found in 50 patients. Of 122 patients with an injury limited to the non-tooth-bearing facial skeleton, 12 sustained dental trauma (10%). In patients with fractures limited to the maxilla (n = 41), 6 patients had dental injuries (14.5%). In patients with fractures to the mandible (n = 61), 24 sustained dental injuries (39%). When we compared the type of tooth lesion and the location, simple crown fractures prevailed in both jaws. Patients with a fracture of the mandible were most likely to have a dental injury (39.3%). The highest incidence of dental lesions was found in the maxilla in combination with fractures of the lower jaw (39%). This incidence was even higher than the incidence of dental lesions in the lower jaw in combination with fractures of the mandible (24%). Knowledge of the association of dental injuries and maxillofacial fractures is a basic tool for their prevention. Our study showed that in cases of trauma with mandibular fracture, the teeth in the upper jaw might be at higher risk than the teeth in the lower jaw. Further larger-scale studies on this topic could clarify this finding and may provide suggestions for the amelioration of safety devices (such as modified bicycle helmets).


Assessment of occupational noise-related hearing impairment among dental health personnel.
The purpose of the study was to examine hearing thresholds among dental personnel. The secondary aim was to evaluate sound levels among dental equipment that dental personnel are exposed to. Two hundred forty-four dentists, dental technicians, dental assistants, and dental students participated. Sixty-two participated as a control group. Audiological thresholds for the test groups were compared to the control group. All participants were from Jordan University Hospital. Participants completed a questionnaire in addition to their audiometric testing. Otoscopy, tympanometry, and pure tone audiometry were included in their assessment. Three-factor ANOVA and t tests were utilized to assess the statistical differences of hearing thresholds among the groups and between the two ears. Pearson correlation test was used to assess the effect of age, experience, and duration of exposure on the degree of hearing loss in the test groups for both ears. The authors reported statistically significant differences among hearing thresholds between the control group and others. Left hearing thresholds were noted to be significantly poorer in the left versus right ear at 1000, 2000, 4000, and 8000 Hz in dental assistants. The authors also reported a significant relationship between the degree of hearing impairment among dental assistants and the daily duration of exposure to dental occupational noise, followed by age. Hearing impairment was higher among dental professionals than the control group and especially among dental assistants and technicians. The authors recommended screening guidelines and adapting hearing protection methods for dental professionals and particularly for dental assistants and technicians.


Rambam Hospital is the Birthplace of the Modern Version of Transvaginal Ultrasound.
The worldwide use of the transvaginal scanning route has revolutionized obstetrical and gynecologic imaging. The long, slow, and at times challenging aspects of its acceptance by the obstetrical and gynecologic community are the subject of this article. From its inception to its recent use, the dedicated doctors in the Department of Obstetrics and Gynecology at Rambam Medical Center, Haifa, Israel, were instrumental in conceiving and then collaborating with an Israeli manufacturer in the construction and worldwide use of the transvaginal ultrasound probe, resulting in the now well-known field of transvaginal sonography.


"I Have Remained Strong Because of That Food": Acceptability and Use of Lipid-Based Nutrient Supplements Among Pregnant HIV-Infected Ugandan Women Receiving Combination Antiretroviral Therapy.
We evaluated the acceptability and use of macronutrient supplementation among HIV-infected pregnant Ugandan women receiving antiretroviral therapy in a clinical study (NCT 00993031). We first conducted formative research among 56 pregnant and lactating women to select a supplement regimen. Acceptability and use of the supplementation regimen (35 sachets of lipid-based nutrient supplements (LNS) and 4 or 6 kg of instant soy porridge for the household provided monthly) were evaluated among 87 pregnant women. Organoleptic assessments of LNS were favorable. Participants reported consuming LNS a mean of 6.1 days per week, and adherence to recommended consumption behaviors (e.g. frequency, quantity, not sharing) was >80 %. Few women reported negative social consequences of supplementation. The majority of participants also consumed most of the porridge intended for the household. In sum, LNS was acceptable and used regularly. Larger studies to evaluate physical and psychosocial consequences of LNS during pregnancy among HIV-infected women are warranted.


Importance of Th22 Cell Disequilibrium in Immune Thrombocytopenic Purpura.
BACKGROUND The disequilibrium of T helper (Th) cells play an important role in the occurrence and development of immune thrombocytopenic purpura (ITP). Th22 cells, as a newly discovered subset of T lymphocytes, plays an important role in autoimmune disorders and inflammatory diseases. MATERIAL AND METHODS This study explored the role of different lymphocyte subsets in chronic ITP. To explore the value of Th22 cells in the diagnosis of ITP, the numbers of Th1, Th17, and Th22 cells were detected by a 4-color flow cytometric in 32 chronic ITP patients and 30 healthy controls. RESULTS Our data showed that, compared with healthy controls, the numbers of circulating Th1, Th17, and Th22 (p<0.05) cells increased significantly in ITP patients, and Th22 cells were correlated positively with Th1 cells (r=0.4041, p<0.01) and Th17 cells (r=0.4637, p<0.05). Moreover, a positive relationship was found between Th1/Th22 cells and Th1 cells (r=0.7696, p<0.001). CONCLUSIONS A disequilibrium expression profile of Th22 cells in peripheral blood was associated with pathogenesis of ITP, possibly through cooperatively working with Th17 and Th1, which may provide a novel approach for diagnosis of ITP.


Xylitol, mannitol and maltitol as potential sucrose replacers in burger buns.
Burger buns are a source of added sugar, containing 7-12%, in order to ensure their unique texture and taste. Hence, suitable sugar substitutes for burger buns are urgently needed. This study aimed to elucidate the effect of three different polyols on dough and product quality of burger buns. Xylitol, mannitol and maltitol were incorporated individually in a burger bun system, by replacing added sucrose by 30%, 50% and 100%. Wheat starch was used to compare the impact of polyols with another non-sweet bulking agent. The effects on dough properties as well as on the burger buns themselves were investigated. Compared to sugar-rich doughs, polyols lowered the fermentation quality, resulting in lower dough development (-37 to -81%) and poorer gaseous release (-62 to -87%). Furthermore, a delay in gluten network development (+50 to +161%) and a decrease in extensibility (-14 to -18%) with increasing concentrations were detected. Interestingly, maltitol and xylitol did not affect the pasting properties, whereas mannitol increased pasting temperature (+15 °C). Moreover, polyols did not influence the viscoelastic properties of the dough. The incorporation of sugar alcohols led to a significant decrease in specific volume (-30 to -48%), and to a harder crumb texture (+135 to +678%). Moreover, the L*-value increased with increasing amount of polyols, resulting in a very pale crust colour. In conclusion, a reduction of 50% added sucrose by polyols was applicable, whereas mannitol was the most suitable sugar replacer amongst the polyols tested.


Use of quantitative real-time PCR for studying the dissemination of Leptospira interrogans in the guinea pig infection model of leptospirosis.
The dynamics of leptospirosis infection have been poorly studied. The purpose of this study was to determine the LD(50), rate of bacterial dissemination, histopathology and antibody responses against leptospira following inoculation with the highly virulent Leptospira interrogans Fiocruz L1-130 strain in a guinea pig model of leptospirosis. Three routes of infection (intraperitoneal, conjunctival and subcutaneous inoculation) were used to establish disease in guinea pigs. The size and kinetics of leptospiral burdens in the blood and tissues of infected animals were determined over a 1 week course of infection using quantitative real-time PCR (qPCR). Bacteraemia peaked at day 5 post-infection reaching more than 5x10(4) leptospires ml(-1). The highest spirochaetal load was found in the liver and kidneys, and was associated with alterations in organ tissues and a decline in liver and kidney functions. In contrast, lesions and bacteria were not detected in guinea pigs infected with an avirulent strain derived from a high-passage-number in vitro-passaged variant of the Fiocruz L1-130 strain. The use of qPCR supports the findings of earlier studies and provides an easy and reliable method for the quantification of L. interrogans in the tissues of infected animals. qPCR will be used in future studies to evaluate the efficacy of vaccine candidates against leptospirosis and the virulence of selected L. interrogans mutants relative to the parental strain.


Mislocalized opsin and cAMP signaling: a mechanism for sprouting by rod cells in retinal degeneration.
In human retinal degeneration, rod photoreceptors reactively sprout neurites. The mechanism is unknown in part because of the paucity of animal models displaying this feature of human pathology. We tested the role of cAMP and opsin in sprouting by tiger salamander rod cells, photoreceptors that can produce reactive growth. In vitro systems of isolated photoreceptor cells and intact neural retina were used. cAMP signaling was manipulated with nucleotide analogues, enzyme stimulators, agonists for adenosine and dopamine receptors, and the opsin agonist, β-ionone. Levels of cAMP were determined by radioimmunoassay, and protein levels by Western blot and quantitative immunocytochemistry. Neuritic growth was assayed by image analysis and conventional and confocal microscopy. cAMP analogues and stimulation of adenylyl cyclase (AC) directly or through G-protein-coupled receptors resulted in significant increases in neuritic growth of isolated rod, but not cone, cells. The signaling pathway included protein kinase A (PKA) and phosphorylation of the transcription factor cAMP response element-binding protein (pCREB). Opsin, a G-linked receptor, is present throughout the plasmalemma of isolated cells; its activation also induced sprouting. In neural retina, rod sprouting was significantly increased by β-ionone with concomitant increases in cAMP, pCREB, and synaptic proteins. Notably, opsin stimulated sprouting only when mislocalized to the plasmalemma of the rod cell body. cAMP causes neuritic sprouting in rod, but not cone, cells through the AC-PKA-CREB pathway known to be associated with synaptic plasticity. We propose that in retinal disease, mislocalized rod opsin gains access to cAMP signaling, which leads to neuritic sprouting.


Improved algorithm for simulations of divergent-light halos.
Divergent-light halos are produced when light from nearby light sources is scattered by ice crystals in the atmosphere. We present a theory of divergent-light halos leading to an improved algorithm for the simulation of such halos. Contrary to the algorithm that we presented earlier for simulating such halos, the new algorithm includes a mathematically rigorous weighting of the events. The computer implementation is very compact, and the whole procedure is elegant and conceptually easy to understand. We also present a new simulation atlas showing halos produced by crystals of different shapes and orientations for a set of elevations of the light source.


Lack of association of a variable number of aspartic acid residues in the asporin gene with osteoarthritis susceptibility: case-control studies in Spanish Caucasians.
A recent genetic association study has identified a microsatellite in the coding sequence of the asporin gene as a susceptibility factor for osteoarthritis (OA). Alleles of this microsatellite determine the variable number of aspartic acid residues in the amino-terminal end of the asporin protein. Asporin binds directly to the growth factor transforming growth factor beta and inhibits its anabolic effects in cartilage, which include stimulation of collagen and aggrecan synthesis. The OA-associated allele, with 14 aspartic acid residues, inhibits the anabolic effects of transforming growth factor beta more strongly than other asporin alleles, leading to increased OA liability. We have explored whether the association found in several cohorts of Japanese hip OA and knee OA patients was also present in Spanish Caucasians. We studied patients that had undergone total joint replacement for primary OA in the hip (n = 303) or the knee (n = 188) and patients with hand OA (n = 233), and we compared their results with controls (n = 294) lacking overt OA clinical symptoms. No significant differences were observed in any of the multiple comparisons performed, which included global tests of allele frequency distributions and specific comparisons as well as stratification by affected joint and by sex. Our results, together with reports from the United Kingdom and Greece, indicate that the stretch of aspartic acid residues in asporin is not an important factor in OA susceptibility among European Caucasians. It remains possible that lifestyle, environmental or genetic differences allow for an important effect of asporin variants in other ethnic groups as has been reported in the Japanese, but this should be supported by additional studies.


ASGE Bariatric Endoscopy Task Force systematic review and meta-analysis assessing the ASGE PIVI thresholds for adopting endoscopic bariatric therapies.
The increasing global burden of obesity and its associated comorbidities has created an urgent need for additional treatment options to fight this pandemic. Endoscopic bariatric therapies (EBTs) provide an effective and minimally invasive treatment approach to obesity that would increase treatment options beyond surgery, medications, and lifestyle measures. This systematic review and meta-analysis were performed by the American Society for Gastrointestinal Endoscopy (ASGE) Bariatric Endoscopy Task Force comprising experts in the subject area and the ASGE Technology Committee Chair to specifically assess whether acceptable performance thresholds outlined by an ASGE Preservation and Incorporation of Valuable endoscopic Innovations (PIVI) document for clinical adoption of available EBTs have been met. After conducting a comprehensive search of several English-language databases, we performed direct meta-analyses by using random-effects models to assess whether the Orbera intragastric balloon (IGB) (Apollo Endosurgery, Austin, Tex) and the EndoBarrier duodenal-jejunal bypass sleeve (DJBS) (GI Dynamics, Lexington, Mass) have met the PIVI thresholds. The meta-analyses results indicate that the Orbera IGB meets the PIVI thresholds for both primary and nonprimary bridge obesity therapy. Based on a meta-analysis of 17 studies including 1683 patients, the percentage of excess weight loss (%EWL) with the Orbera IGB at 12 months was 25.44% (95% confidence interval [CI], 21.47%-29.41%) (random model) with a mean difference in %EWL over controls of 26.9% (95% CI, 15.66%-38.24%; P ≤ .01) in 3 randomized, controlled trials. Furthermore, the pooled percentage of total body weight loss (% TBWL) after Orbera IGB implantation was 12.3% (95% CI, 7.9%–16.73%), 13.16% (95% CI, 12.37%–13.95%), and 11.27% (95% CI, 8.17%–14.36%) at 3, 6, and 12 months after implantation, respectively, thus exceeding the PIVI threshold of 5% TBWL for nonprimary (bridge) obesity therapy. With the data available, the DJBS liner does appear to meet the %EWL PIVI threshold at 12 months, resulting in 35% EWL (95% CI, 24%-46%) but does not meet the 15% EWL over control required by the PIVI. We await review of the pivotal trial data on the efficacy and safety of this device. Data are insufficient to evaluate PIVI thresholds for any other EBT at this time. Both evaluated EBTs had ≤5% incidence of serious adverse events as set by the PIVI document to indicate acceptable safety profiles. Our task force consequently recognizes the Orbera IGB for meeting the PIVI criteria for the management of obesity. As additional data from the other EBTs become available, we will update our recommendations accordingly.


Haploidentical hematopoietic transplantation without T-cell depletion: current status and future perspectives.
Human leukocyte antigen (HLA)-haploidentical hematopoietic stem cell transplantation (HLA-haplo HSCT) without T-cell depletion has tremendously progressed over the past 20 years and has become a feasible treatment option for leukemia patients without an HLA-identical sibling donor. Advances in conditioning regimens, graft manipulation, and pharmacological graft-versus-host disease (GVHD) prophylaxis have reduced the risk of fatal graft failure and severe GVHD, two of the most serious complications of traversing the HLA barrier. According to clinical observations, killer immunoglobulin-like receptor (KIR) mismatch and donor-specific anti-HLA (DSA) antibodies-negative status play potential roles in reducing the risk of GVHD and graft failure following HLA-haploidentical SCT. New strategies to improve transplant outcomes include donor lymphocyte, NK cell and selected T-cell subset infusion, mesenchymal stem cell (MSC) co-transplantation and interleukin-2 (IL-2) application. Future challenges remain in improving post-transplant immune reconstitution and finding the best approach to reduce the incidence and severity of GVHD while simultaneously preserving the graft-versus leukemia effect to prevent the recurrence of underlying malignancy.


Size and edge dependence of two-photon absorption in rectangular graphene quantum dots.
The size and edge-dependence of two-photon absorption (TPA) for rectangular graphene quantum dots (GQDs) is investigated theoretically in the framework of Dirac equation under hard wall boundary conditions. The TPA cross section associated with interband transitions around K point is derived and the transition selection rules are obtained. Results reveal that when the size of zigzag-edge M = 3M0 ± 1 (M0 is an integer), the GQD exhibits a semiconductor while for M = 3M0 it is metallic. For semiconducting rectangular GQDs, TPA is tuned by the sizes of both edges in GQDs and the armchair-edge dimension contributes more to TPA. While for metallic rectangular GQDs, zigzag-edge dimension affects TPA little and the position of absorption peak and the magnitude of the TPA coefficient are determined by the size of armchair-edge and the resonant enhancement occurs.


Aloe-Emodin Induces Endoplasmic Reticulum Stress-Dependent Apoptosis in Colorectal Cancer Cells.
BACKGROUND Recently, colorectal cancer has become a more common type of tumor in the world. Research has reported that several kinds of single compounds of Chinese herbs have shown anti-tumor activity in colorectal cancer. Aloe-emodin (AE), a natural compound extract from Aloe Vera, has been discovered to suppress cell proliferation and accelerate apoptosis in a variety of tumor cells. Whether AE exerts an effect on colorectal cancer cells has not yet been investigated. MATERIAL AND METHODS In this study, we examined the impact of AE on SW620 and HT29 colorectal cancer cell lines. After treatment with various concentrations of AE (10, 20, and 40 μM), cell proliferation, cell apoptosis, reactive oxygen species (ROS) generation, cytosolic calcium level, and related gene expression were analyzed. RESULTS Our results indicated that AE suppressed cell viability and induced cell apoptosis in SW620 and HT29 cell lines. Furthermore, both cell lines when exposed to AE generated ROS, which induces endoplasmic reticulum (ER) stress. We then detected the expression of ER stress-related proteins and cytosolic calcium levels. We found that cells exposure to AE had upregulation of unfolded protein response (UPR) proteins like glucose-related protein 78 (GRP78), phosphorylated protein kinase R (PKR)-like ER kinase (p-PERK), phosphorylated eukaryotic initiation factor-2α (p-eIF2α), and transcription factor C/EBP homologous protein (CHOP). Meanwhile, we detected an increased cytosolic calcium content followed by the upregulation of the calpain-1, calpain-2 and caspase-12. CHOP and caspase-12 are important regulatory factors leading to cell apoptosis. CONCLUSIONS AE might serve as a candidate in the treatment of colorectal cancer through inducing ER stress-dependent apoptosis.


New technologies to improve healthcare in low- and middle-income countries: Global Grand Challenges satellite event, Oxford University Clinical Research Unit, Ho Chi Minh City, 17th-18th September 2019.
We report the outputs of a satellite event in Ho Chi Minh City, Vietnam, organized as part of the "2 nd Global Grand Challenges of Engineering Summit". The event considered challenges and potential solutions for improving low- and middle-income country (LMIC) healthcare systems, with particular reference to critical care. Participants from key regional and local stakeholders in healthcare and engineering discussed how new advances in technology, especially in the field of Artificial Intelligence, could be of potential benefit. This article summarizes the perspectives and conclusions of a group of key stakeholders from LMICs across South and South East Asia.


Current advances in systems and integrative biology.
Systems biology has gained a tremendous amount of interest in the last few years. This is partly due to the realization that traditional approaches focusing only on a few molecules at a time cannot describe the impact of aberrant or modulated molecular environments across a whole system. Furthermore, a hypothesis-driven study aims to prove or disprove its postulations, whereas a hypothesis-free systems approach can yield an unbiased and novel testable hypothesis as an end-result. This latter approach foregoes assumptions which predict how a biological system should react to an altered microenvironment within a cellular context, across a tissue or impacting on distant organs. Additionally, re-use of existing data by systematic data mining and re-stratification, one of the cornerstones of integrative systems biology, is also gaining attention. While tremendous efforts using a systems methodology have already yielded excellent results, it is apparent that a lack of suitable analytic tools and purpose-built databases poses a major bottleneck in applying a systematic workflow. This review addresses the current approaches used in systems analysis and obstacles often encountered in large-scale data analysis and integration which tend to go unnoticed, but have a direct impact on the final outcome of a systems approach. Its wide applicability, ranging from basic research, disease descriptors, pharmacological studies, to personalized medicine, makes this emerging approach well suited to address biological and medical questions where conventional methods are not ideal.


Computer-assisted diagnosis techniques (dermoscopy and spectroscopy-based) for diagnosing skin cancer in adults.
Early accurate detection of all skin cancer types is essential to guide appropriate management and to improve morbidity and survival. Melanoma and cutaneous squamous cell carcinoma (cSCC) are high-risk skin cancers which have the potential to metastasise and ultimately lead to death, whereas basal cell carcinoma (BCC) is usually localised with potential to infiltrate and damage surrounding tissue. Anxiety around missing early curable cases needs to be balanced against inappropriate referral and unnecessary excision of benign lesions. Computer-assisted diagnosis (CAD) systems use artificial intelligence to analyse lesion data and arrive at a diagnosis of skin cancer. When used in unreferred settings ('primary care'), CAD may assist general practitioners (GPs) or other clinicians to more appropriately triage high-risk lesions to secondary care. Used alongside clinical and dermoscopic suspicion of malignancy, CAD may reduce unnecessary excisions without missing melanoma cases. To determine the accuracy of CAD systems for diagnosing cutaneous invasive melanoma and atypical intraepidermal melanocytic variants, BCC or cSCC in adults, and to compare its accuracy with that of dermoscopy. We undertook a comprehensive search of the following databases from inception up to August 2016: Cochrane Central Register of Controlled Trials (CENTRAL); MEDLINE; Embase; CINAHL; CPCI; Zetoc; Science Citation Index; US National Institutes of Health Ongoing Trials Register; NIHR Clinical Research Network Portfolio Database; and the World Health Organization International Clinical Trials Registry Platform. We studied reference lists and published systematic review articles. Studies of any design that evaluated CAD alone, or in comparison with dermoscopy, in adults with lesions suspicious for melanoma or BCC or cSCC, and compared with a reference standard of either histological confirmation or clinical follow-up. Two review authors independently extracted all data using a standardised data extraction and quality assessment form (based on QUADAS-2). We contacted authors of included studies where information related to the target condition or diagnostic threshold were missing. We estimated summary sensitivities and specificities separately by type of CAD system, using the bivariate hierarchical model. We compared CAD with dermoscopy using (a) all available CAD data (indirect comparisons), and (b) studies providing paired data for both tests (direct comparisons). We tested the contribution of human decision-making to the accuracy of CAD diagnoses in a sensitivity analysis by removing studies that gave CAD results to clinicians to guide diagnostic decision-making. We included 42 studies, 24 evaluating digital dermoscopy-based CAD systems (Derm-CAD) in 23 study cohorts with 9602 lesions (1220 melanomas, at least 83 BCCs, 9 cSCCs), providing 32 datasets for Derm-CAD and seven for dermoscopy. Eighteen studies evaluated spectroscopy-based CAD (Spectro-CAD) in 16 study cohorts with 6336 lesions (934 melanomas, 163 BCC, 49 cSCCs), providing 32 datasets for Spectro-CAD and six for dermoscopy. These consisted of 15 studies using multispectral imaging (MSI), two studies using electrical impedance spectroscopy (EIS) and one study using diffuse-reflectance spectroscopy. Studies were incompletely reported and at unclear to high risk of bias across all domains. Included studies inadequately address the review question, due to an abundance of low-quality studies, poor reporting, and recruitment of highly selected groups of participants.Across all CAD systems, we found considerable variation in the hardware and software technologies used, the types of classification algorithm employed, methods used to train the algorithms, and which lesion morphological features were extracted and analysed across all CAD systems, and even between studies evaluating CAD systems. Meta-analysis found CAD systems had high sensitivity for correct identification of cutaneous invasive melanoma and atypical intraepidermal melanocytic variants in highly selected populations, but with low and very variable specificity, particularly for Spectro-CAD systems. Pooled data from 22 studies estimated the sensitivity of Derm-CAD for the detection of melanoma as 90.1% (95% confidence interval (CI) 84.0% to 94.0%) and specificity as 74.3% (95% CI 63.6% to 82.7%). Pooled data from eight studies estimated the sensitivity of multispectral imaging CAD (MSI-CAD) as 92.9% (95% CI 83.7% to 97.1%) and specificity as 43.6% (95% CI 24.8% to 64.5%). When applied to a hypothetical population of 1000 lesions at the mean observed melanoma prevalence of 20%, Derm-CAD would miss 20 melanomas and would lead to 206 false-positive results for melanoma. MSI-CAD would miss 14 melanomas and would lead to 451 false diagnoses for melanoma. Preliminary findings suggest CAD systems are at least as sensitive as assessment of dermoscopic images for the diagnosis of invasive melanoma and atypical intraepidermal melanocytic variants. We are unable to make summary statements about the use of CAD in unreferred populations, or its accuracy in detecting keratinocyte cancers, or its use in any setting as a diagnostic aid, because of the paucity of studies. In highly selected patient populations all CAD types demonstrate high sensitivity, and could prove useful as a back-up for specialist diagnosis to assist in minimising the risk of missing melanomas. However, the evidence base is currently too poor to understand whether CAD system outputs translate to different clinical decision-making in practice. Insufficient data are available on the use of CAD in community settings, or for the detection of keratinocyte cancers. The evidence base for individual systems is too limited to draw conclusions on which might be preferred for practice. Prospective comparative studies are required that evaluate the use of already evaluated CAD systems as diagnostic aids, by comparison to face-to-face dermoscopy, and in participant populations that are representative of those in which the test would be used in practice.


Large eddy simulation of the pharyngeal airflow associated with obstructive sleep apnea syndrome at pre and post-surgical treatment.
Obstructive Sleep Apnea Syndrome (OSAS) is the most common sleep-disordered breathing medical condition and a potentially life-threatening affliction. Not all the surgical or non-surgical OSAS therapies are successful for each patient, also in part because the primary factors involved in the etiology of this disorder are not completely understood. Thus, there is a need for improving both diagnostic and treatment modalities associated with OSAS. A verified and validated (in terms of mean velocity and pressure fields) Large Eddy Simulation approach is used to characterize the abnormal pharyngeal airflow associated with severe OSAS and its interaction with the airway wall in a subject who underwent surgical treatment. The analysis of the unsteady flow at pre- and post-treatment is used to illustrate the airflow dynamics in the airway associated with OSAS and to reveal as well, the changes in the flow variables after the treatment. At pre-treatment, large airflow velocity and wall shear stress values were found at the obstruction site in all cases. Downstream of obstruction, flow separation generated flow recirculation regions and enhanced the turbulence production in the jet-like shear layers. The interaction between the generated vortical structures and the pharyngeal airway wall induced large fluctuations in the pressure forces acting on the pharyngeal wall. After the surgery, the flow field instabilities vanished and both airway resistance and wall shear stress values were significantly reduced.


Fluorine-18 deoxyglucose and false-positive results: a major problem in the diagnostics of oncological patients.
Fluorine-18 deoxyglucose (FDG) is not a very tumour-specific substance, and its accumulation in benign lesions with increased glucose metabolism may give rise to false-positive results and hence cause FDG positron emission tomography (PET) to display relatively low specificity (frequently below 85%). Correct interpretation of FDG PET studies is predicated upon detailed knowledge of morphological abnormalities, and the importance of the correlation of functional and morphological information, as derived from computed tomography or magnetic resonance imaging, is discussed. It is emphasized that image fusion programs cannot substitute for understanding of functional and morphological methods. The reconstruction of PET cross-sections is considered, and it is concluded that an iterative image reconstruction method is to be favoured, given its advantages in reducing image artefacts and improving quantification of radioactivity concentrations. The differentiation of malignant and benign lesions when using FDG PET is then reviewed; false-positive findings may be obtained, for example, in patients with acute inflammatory lesions, chronic pancreatitis, retroperitoneal fibrosis or salivary gland tumours. It is suggested that these problems may be alleviated by means of multitracer studies, e.g. using carbon-11 labelled aminoisobutyric acid for quantification of A-type amino acid transport. Finally, the effects of radiotherapy and chemotherapy on FDG uptake and the problems that accrue from these effects are reviewed. Both radiotherapy and chemotherapy can cause increased FDG uptake, complicating diagnosis and evaluation. Knowledge of the effects of different treatment procedures on regional FDG metabolism is therefore necessary for correct interpretation of the PET data.


Ticlopidine as a selective mechanism-based inhibitor of human cytochrome P450 2C19.
Experiments using recombinant yeast-expressed human liver cytochromes P450 confirmed previous literature data indicating that ticlopidine is an inhibitor of CYP 2C19. The present studies demonstrated that ticlopidine is selective for CYP 2C19 within the CYP 2C subfamily. UV-visible studies on the interaction of a series of ticlopidine derivatives with CYP 2C19 showed that ticlopidine binds to the CYP 2C19 active site with a K(s) value of 2.8 +/- 1 microM. Derivatives that do not involve either the o-chlorophenyl substituent, the free tertiary amine function, or the thiophene ring of ticlopidine did not lead to such spectral interactions and failed to inhibit CYP 2C19. Ticlopidine is oxidized by CYP 2C19 with formation of two major metabolites, the keto tautomer of 2-hydroxyticlopidine (1) and the dimers of ticlopidine S-oxide (TSOD) (V(max) = 13 +/- 2 and 0.4 +/- 0.1 min(-1)). During this oxidation, CYP 2C19 was inactivated; the rate of its inactivation was time and ticlopidine concentration dependent. This process meets the chemical and kinetic criteria generally accepted for mechanism-based enzyme inactivation. It occurs in parralel with CYP 2C19-catalyzed oxidation of ticlopidine, is inhibited by an alternative well-known substrate of CYP 2C19, omeprazole, and correlates with the covalent binding of ticlopidine metabolite(s) to proteins. Moreover, CYP 2C19 inactivation is not inhibited by the presence of 5 mM glutathione, suggesting that it is due to an alkylation occurring inside the CYP 2C19 active site. The effects of ticlopidine on CYP 2C19 are very analogous with those previously described for the inactivation of CYP 2C9 by tienilic acid. This suggests that a similar electrophilic intermediate, possibly a thiophene S-oxide, is involved in the inactivation of CYP 2C19 and CYP 2C9 by ticlopidine and tienilic acid, respectively. The kinetic parameters calculated for ticlopidine-dependent inactivation of CYP 2C19, i.e., t(1/2max) = 3.4 min, k(inact) = 3.2 10(-3) s(-1), K(I) = 87 microM, k(inact)/K(I) = 37 L.mol(-1).s(-1), and r (partition ratio) = 26 (in relation with formation of 1 + TSOD), classify ticlopidine as an efficient mechanism-based inhibitor although somewhat less efficient than tienilic acid for CYP 2C9. Importantly, ticlopidine is the first selective mechanism-based inhibitor of human liver CYP 2C19 and should be a new interesting tool for studying the topology of the active site of CYP 2C19.


Phosphoryl transfer and calcium ion occlusion in the calcium pump.
A tight coupling between adenosine triphosphate (ATP) hydrolysis and vectorial ion transport has to be maintained by ATP-consuming ion pumps. We report two crystal structures of Ca2+-bound sarco(endo)plasmic reticulum Ca2+-adenosine triphosphatase (SERCA) at 2.6 and 2.9 angstrom resolution in complex with (i) a nonhydrolyzable ATP analog [adenosine (beta-gamma methylene)-triphosphate] and (ii) adenosine diphosphate plus aluminum fluoride. SERCA reacts with ATP by an associative mechanism mediated by two Mg2+ ions to form an aspartyl-phosphorylated intermediate state (Ca2-E1 approximately P). The conformational changes that accompany the reaction with ATP pull the transmembrane helices 1 and 2 and close a cytosolic entrance for Ca2+, thereby preventing backflow before Ca2+ is released on the other side of the membrane.


Internet surveillance: content analysis and monitoring of product-specific internet prescription opioid abuse-related postings.
This study describes the development of a systematic approach to the analysis of Internet chatter as a means of monitoring potentially abusable opioid analgesics. Message boards dedicated to drug abuse were selected using specific inclusion criteria. Threaded discussions containing 48,293 posts were captured. A coding system was created to compare content of posts related to 3 opioid analgesics: Kadian, Vicodin, and OxyContin. The number of posts containing mentions of the target drugs were significantly different [OxyContin (1813)>Vicodin (940)>Kadian (27), P<0.001]. Analyses revealed that these differences were not simply a reflection of the availability of each product (ie, number of prescriptions written). Reliability tests indicated that the content coding system achieved good interrater reliability coefficients (average kappa across all categories=0.76, range=0.52 to 1.0). Content analysis of a sample of 234 randomly selected posts indicated that the proportion of Internet posts endorsing abuse of Kadian was statistically significantly less than OxyContin (45.5% vs. 68.4%, P=0.036, not adjusted for multiple comparisons). These results suggest that a systematic approach to postmarketing surveillance of Internet chatter related to pharmaceutical products is feasible and yields reliable information about the quantity of discussion of specific products and qualitative information regarding the nature of the discussions. Kadian was associated with fewer Internet mentions than either OxyContin or Vicodin. This investigation stands as a first attempt to establish systematic methods for conducting Internet surveillance.


Risk factors associated with low birth weight of neonates among pregnant women attending a referral hospital in northern Tanzania.
According to the World Health Organization low birth weight (LBW) babies are those born with less than 2500g. A descriptive retrospective cross - sectional study using existing data from a one-year (2001) block of birth registers of 3464 pregnant women was done at Kilimanjaro Christian Medical Centre in Moshi, Tanzania. The objective was to determine factors associated with LBW and their contribution to the problem. Out of 648 pregnant women who were tested for HIV infection 59 (9.1%) were positive for the infection. Twelve (20.3%) of HIV positive women gave birth to LBW neonates. HIV positive women were twice more likely to give birth to LBW infants than HIV negative ones (chi2 = 6.7; P < 0.01; OR = 2.4; 1.1, 5.1). Mothers without formal education were 4 times more likely to give birth to LBW neonates than those who had attained higher education (OR = 3.6; 2.2, 5.9). There was a linear decrease in low birth weights of newborns as fraternal educational level increased (chi2 for linear trend = 42.7; P < 0.01). There was no statistically significant difference among parents' occupations regarding LBW of their newborns. Unmarried mothers were more likely to give birth to LBW neonates as compared to their married counterparts (OR = 1.65; 1.2, 2.2) and the difference was statistically significant (chi2 = 13.0, P < 0.01). Hypertension, pre-eclampsia and eclampsia disease complex had the highest prevalence (46.67%) and population attributable fraction of low birth weight (PAF = 25.2%; CI = 22.0-27.6). Bleeding and schistosomiasis had the same prevalence (33.33%) of LBW babies. Other complications and diseases which contributed to high prevalence of LBW included anaemia (25%), thromboembolic diseases (20%), tuberculosis (17%) and malaria (14.8%). Prevalence of LBW was high in women with premature rupture of membrane (38%), placenta previa (17%) and abruption of placenta (15.5%). LBW was strongly associated with gestational age below 37 weeks (OR = 2; CI = 1.5, 2.8) contributing to 42% of LBW deliveries in the study population (PAF = 42.4%: 25, 55). Pregnant women with malnutrition (BMI < 18) gave the highest proportions 17% of LBW children followed by underweight (BMI; 18-22) who gave 15.5% of LBW neonates. There was a statistical significant difference between the proportions of LBW infants from mothers who did not receive antenatal care (28.6%) and those who attended for the services (13.8%) (chi2 = 8.8; P = 0.01). There is need of increasing promotion of reproductive health services in relation to safe motherhood at community level in order to reduce risk factors of LBW.


Perceived accessibility of cigarettes among youth: a prospective cohort study.
The accessibility of tobacco for youth is difficult to measure, partly because of the varied sources of cigarettes. Perceptions about the accessibility of cigarettes assesses availability from all potential sources and has been found to predict future smoking. This study examines the determinants of perceived accessibility from the perspective of a longitudinal study. Data were derived from the second Development and Assessment of Nicotine Dependence in Youth study, a 4-year longitudinal study of 1246 sixth-grade students who underwent up to 11 in-person interviews from 2002 to 2006. Perceived accessibility was assessed prospectively by asking students whether they agreed or disagreed with the statement It would be easy for me to get a cigarette. At baseline, the average age was 12 years; 85% had never before smoked; more than 21% perceived that cigarettes were easy to obtain. Perceived accessibility increased over the 4 years among the cohort. Youth with friends who smoked or whose parents allowed the watching of R-rated movies were more likely to perceive easy accessibility of cigarettes. Among nonsmokers, there was a dose-response relationship between perceived accessibility and exposure to smoking in the family environment. Nonsmokers who knew commercial sources of cigarettes also had higher perceived accessibility. Several potentially preventable exposures to tobacco in the family and social environment contribute to confidence among some youth that cigarettes are easy to obtain. Further studies are needed to determine if modifying these factors (e.g., through programs to reduce the exposure of youth to tobacco products in the home) might reduce perceived accessibility.


Competition for hydrogen-bond formation in the helix-coil transition and protein folding.
The problem of the helix-coil transition of biopolymers in explicit solvents, such as water, with the ability for hydrogen bonding with a solvent is addressed analytically using a suitably modified version of the Generalized Model of Polypeptide Chains. Besides the regular helix-coil transition, an additional coil-helix or reentrant transition is also found at lower temperatures. The reentrant transition arises due to competition between polymer-polymer and polymer-water hydrogen bonds. The balance between the two types of hydrogen bonding can be shifted to either direction through changes not only in temperature, but also by pressure, mechanical force, osmotic stress, or other external influences. Both polypeptides and polynucleotides are considered within a unified formalism. Our approach provides an explanation of the experimental difficulty of observing the reentrant transition with pressure and underscores the advantage of pulling experiments for studies of DNA. Results are discussed and compared with those reported in a number of recent publications with which a significant level of agreement is obtained.


Kassiopeia: a database and web application for the analysis of mutually exclusive exomes of eukaryotes.
Alternative splicing is an important process in higher eukaryotes that allows obtaining several transcripts from one gene. A specific case of alternative splicing is mutually exclusive splicing, in which exactly one exon out of a cluster of neighbouring exons is spliced into the mature transcript. Recently, a new algorithm for the prediction of these exons has been developed based on the preconditions that the exons of the cluster have similar lengths, sequence homology, and conserved splice sites, and that they are translated in the same reading frame. In this contribution we introduce Kassiopeia, a database and web application for the generation, storage, and presentation of genome-wide analyses of mutually exclusive exomes. Currently, Kassiopeia provides access to the mutually exclusive exomes of twelve Drosophila species, the thale cress Arabidopsis thaliana, the flatworm Caenorhabditis elegans, and human. Mutually exclusive spliced exons (MXEs) were predicted based on gene reconstructions from Scipio. Based on the standard prediction values, with which 83.5% of the annotated MXEs of Drosophila melanogaster were reconstructed, the exomes contain surprisingly more MXEs than previously supposed and identified. The user can search Kassiopeia using BLAST or browse the genes of each species optionally adjusting the parameters used for the prediction to reveal more divergent or only very similar exon candidates. We developed a pipeline to predict MXEs in the genomes of several model organisms and a web interface, Kassiopeia, for their visualization. For each gene Kassiopeia provides a comprehensive gene structure scheme, the sequences and predicted secondary structures of the MXEs, and, if available, further evidence for MXE candidates from cDNA/EST data, predictions of MXEs in homologous genes of closely related species, and RNA secondary structure predictions. Kassiopeia can be accessed at http://www.motorprotein.de/kassiopeia.


A novel pro-apoptotic function of RACK1: suppression of Src activity in the intrinsic and Akt pathways.
Earlier we showed that RACK1 regulates growth of human colon cells by suppressing Src activity at G(1) and mitotic checkpoints. Here, we show that RACK1 also induces apoptosis of the cells, partly by inhibiting Src. In the intrinsic pathway, RACK1 inhibits expression of anti-apoptotic Bcl-2 and Bcl-X(L), induces expression of pro-apoptotic Bim, targets Bim and Bax to the mitochondria, induces oligomerization of Bax (which requires Bim and inhibition of Src), depolarizes mitochondria membranes, releases cytochrome c, and activates caspases-9 and -3 and death substrates. Bax and Bim are required for RACK1-mediated mitochondrial cell death. RACK1-induced oligomerization of Bax is required for staurosporine-mediated cell death. RACK1 also induces apoptosis by blocking Src activation of the Akt cell survival pathway. This leads to activation of the transcription factor FOXO3, a potent inducer of apoptosis and G(1) arrest. Collectively, our results show that RACK1, partly by inhibiting Src, promotes mitochondrial cell death and blocks Akt-mediated cell survival. Thus, RACK1 inhibits growth and induces death of colon cells. Exploitation of these dual functions could lead to novel colon cancer therapies that mimic RACK1 function.


The role of perlecan and endorepellin in the control of tumor angiogenesis and endothelial cell autophagy.
During tumor growth and angiogenesis there is a dynamic remodeling of tissue architecture often accompanied by the release of extracellular matrix constituents full of biological activity. One of the key constituents of the tumor microenvironment is the large heparan sulfate proteoglycan perlecan. This proteoglycan, strategically located at cell surfaces and within basement membranes, is a well-defined pro-angiogenic molecule when intact. However, when partially processed by proteases released during cancer remodeling and invasion, the C-terminal fragment of perlecan, known as endorepellin, has opposite effects than its parent molecule. Endorepellin is a potent inhibitor of angiogenesis by exerting a dual receptor antagonism by simultaneously engaging VEGFR2 and α2β1 integrin. Signaling through the α2β1 integrin leads to actin disassembly and block of endothelial cell migration, necessary for capillary morphogenesis. Signaling through the VEGFR2 induces dephosphorylation of the receptor via activation of SHP-1 and suppression of downstream proangiogenic effectors, especially attenuating VEGFA expression. A novel and emerging role of endorepellin is its ability to evoke autophagy by activating Peg3 and various canonical autophagic markers. This effect is specific for endothelial cells as these are the primary cells expressing both VEGFR2 and α2β1 integrin. Thus, an endogenous fragment of a ubiquitous proteoglycan can regulate both angiogenesis and autophagy through a dual receptor antagonism. The biological properties of this natural endogenous protein place endorepellin as a potential therapeutic agent against cancer or diseases where angiogenesis is prominent.


Human Activity Recognition Models in Ontology Networks.
We present Arianna+, a framework to design networks of ontologies for representing knowledge enabling smart homes to perform human activity recognition online. In the network, nodes are ontologies allowing for various data contextualisation, while edges are general-purpose computational procedures elaborating data. Arianna+ provides a flexible interface between the inputs and outputs of procedures and statements, which are atomic representations of ontological knowledge. Arianna+ schedules procedures on the basis of events by employing logic-based reasoning, that is, by checking the classification of certain statements in the ontologies. Each procedure involves input and output statements that are differently contextualized in the ontologies based on specific prior knowledge. Arianna+ allows to design networks that encode data within multiple contexts and, as a reference scenario, we present a modular network based on a spatial context shared among all activities and a temporal context specialized for each activity to be recognized. In the article, we argue that a network of small ontologies is more intelligible and has a reduced computational load than a single ontology encoding the same knowledge. Arianna+ integrates in the same architecture heterogeneous data processing techniques, which may be better suited to different contexts. Thus, we do not propose a new algorithmic approach to activity recognition, instead, we focus on the architectural aspects for accommodating logic-based and data-driven activity models in a context-oriented way. Also, we discuss how to leverage data contextualization and reasoning for activity recognition, and to support an iterative development process driven by domain experts.


Postural Stability When Walking and Exposed to Mediolateral Oscillatory Motion: Effect of Oscillation Waveform.
Postural stability can be threatened by the low-frequency motions in transport that are usually quantified by their root-mean-square (r.m.s.) acceleration. This study investigated how the stability of walking people depends on the waveform of 1- and 2-Hz mediolateral oscillations of the surface on which they walk. Walking on a treadmill, 20 subjects were perturbed by random oscillations of the treadmill with one-third octave bandwidths: different waveforms with the same r.m.s. acceleration and different waveforms with the same peak acceleration. Stability was measured subjectively and objectively by the velocity of the center of pressure in the mediolateral direction. Subjective and objective measures of walking instability increased with increasing r.m.s. acceleration of oscillations having the same peak acceleration. These same measures of instability were also affected by the peak acceleration when the r.m.s. magnitude of the oscillations was constant, especially with 1-Hz oscillations. It is concluded that r.m.s. measures of acceleration are insufficient to predict the postural stability of walking passengers exposed to mediolateral oscillations and that peaks in the oscillations should also be taken into account.


Coronary microvascular abnormalities in the hypertensive-diabetic rat. A primary cause of cardiomyopathy?
The authors have continued their investigation of the hypertensive-diabetic (HD) rat by evaluating changes in the myocardial microvasculature in this model. Perfusion of HD animals in vivo with a silicone rubber solution revealed numerous areas of microvascular tortuosity, focal constrictions, and microaneurysm formation. These alterations were present to a lesser extent in normoglycemic hypertensive (H) rats, and were distinctly rare in normotensive diabetic rats and unaffected control animals. Quantitation of these vascular lesions revealed highly significant differences between HD animals and the other three groups, with hypertensive rats intermediate between HD rats and diabetic control rats. Areas of pronounced arteriolar constriction were also identified in the HD and H animals with the use of serial sections of Epon-embedded myocardium. It is believed that these lesions represent dynamic changes in the microcirculation, which may cause segmental reperfusion injury to the myocardium, leading to focal replacement fibrosis. Interstitial scarring may result from increased leakiness of small vessels exacerbated by the combined disease. The authors propose that the additive effects of hypertension and diabetes mellitus on the myocardial microcirculation may be a primary cause of cardiomyopathy in this model of human disease.


Diversity of Pneumocystis jirovecii Across Europe: A Multicentre Observational Study.
Pneumocystis jirovecii is an airborne human-specific ascomycetous fungus responsible for Pneumocystis pneumonia (PCP) in immunocompromised patients, affecting >500,000 patients per year (www.gaffi.org). The understanding of its epidemiology is limited by the lack of standardised culture. Recent genotyping data suggests a limited genetic diversity of P. jirovecii. The objective of the study was to assess the diversity of P. jirovecii across European hospitals and analyse P. jirovecii diversity in respect to clinical data obtained from the patients. Genotyping was performed using six already validated short tandem repeat (STR) markers on 249 samples (median: 17 per centre interquartile range [11-20]) from PCP patients of 16 European centres. Mixtures of STR markers (i.e., ≥2 alleles for ≥1 locus) were detected in 67.6% (interquartile range [61.4; 76.5]) of the samples. Mixture was significantly associated with the underlying disease of the patient, with an increased proportion in HIV patients (78.3%) and a decreased proportion in renal transplant recipients (33.3%) (p<0.001). The distribution of the alleles was significantly different (p<0.001) according to the centres in three out of six markers. In analysable samples, 201 combinations were observed corresponding to 137 genotypes: 116 genotypes were country-specific; 12 in two; six in three; and two in four and one in five countries. Nine genotypes were recorded more than once in a given country. Genotype 123 (Gt123) was significantly associated with France (14/15, p<0.001) and Gt16 with Belgium (5/5, p<0.001). More specifically, Gt123 was observed mainly in France (14/15/16 patients) and in renal transplant patient (13/15). Our study showed the wide population diversity across Europe, with evidence of local clusters of patients harbouring a given genotype. These data suggest a specific association between genotype and underlying disease, with evidence of a different natural history of PCP in HIV patients and renal transplant recipients.


The complexity of gene expression dynamics revealed by permutation entropy.
High complexity is considered a hallmark of living systems. Here we investigate the complexity of temporal gene expression patterns using the concept of Permutation Entropy (PE) first introduced in dynamical systems theory. The analysis of gene expression data has so far focused primarily on the identification of differentially expressed genes, or on the elucidation of pathway and regulatory relationships. We aim to study gene expression time series data from the viewpoint of complexity. Applying the PE complexity metric to abiotic stress response time series data in Arabidopsis thaliana, genes involved in stress response and signaling were found to be associated with the highest complexity not only under stress, but surprisingly, also under reference, non-stress conditions. Genes with house-keeping functions exhibited lower PE complexity. Compared to reference conditions, the PE of temporal gene expression patterns generally increased upon stress exposure. High-complexity genes were found to have longer upstream intergenic regions and more cis-regulatory motifs in their promoter regions indicative of a more complex regulatory apparatus needed to orchestrate their expression, and to be associated with higher correlation network connectivity degree. Arabidopsis genes also present in other plant species were observed to exhibit decreased PE complexity compared to Arabidopsis specific genes. We show that Permutation Entropy is a simple yet robust and powerful approach to identify temporal gene expression profiles of varying complexity that is equally applicable to other types of molecular profile data.


Low-dose menaquinone-7 supplementation improved extra-hepatic vitamin K status, but had no effect on thrombin generation in healthy subjects.
Vitamin K is required for the carboxylation of Gla-proteins in the liver (coagulation factors) and extra-hepatic tissues, such as bone (osteocalcin, OC), and arterial wall (matrix Gla-protein, MGP). Although the coagulation factors are essentially fully carboxylated under normal conditions, 10-40 % of OC and MGP remains undercarboxylated. We were therefore interested to study the dose-response effects of extra intake of menaquinones on the carboxylation of the extra-hepatic Gla-proteins. A total of forty-two healthy Dutch men and women aged between 18 and 45 years were randomised into seven groups to receive: placebo capsules or menaquinone-7 (MK-7) capsules at a daily dose of 10, 20, 45, 90, 180 or 360 μg. Circulating uncarboxylated OC (ucOC), carboxylated OC (cOC) and desphospho-uncarboxylated MGP were measured by ELISA. The ucOC:cOC ratio was calculated from circulating ucOC and cOC values. Endogenous thrombin potential and peak height were determined by calibrated automated thrombography. To increase the statistical power, we collapsed the treatment groups into three dosage groups: placebo, low-dose supplementation (doses below RDA, Commission Directive 2008/100/EC), and high-dose supplementation (doses around RDA, Commission Directive 2008/100/EC). MK-7 supplementation at doses in the order of the RDA (Commission Directive 2008/100/EC) increased the carboxylation of circulating OC and MGP. No adverse effects on thrombin generation were observed. Extra MK-7 intake at nutritional doses around the RDA (Commission Directive 2008/100/EC) improved the carboxylation of the extra-hepatic vitamin K-dependent proteins. Whether this improvement contributes to public health, i.e. increasing the protection against age-related diseases needs further investigation in specifically designed intervention trials.


Distinctive patterns of memory function in subgroups of females with Turner syndrome: evidence for imprinted loci on the X-chromosome affecting neurodevelopment.
X-monosomy is a form of Turner syndrome (TS) in which an entire X chromosome is missing. It is usually assumed that neuropsychological deficits in females with TS result from insufficient dosage of gene products from alleles on the sex chromosomes. If so, then parental origin of the single X chromosome should be immaterial. However, if there are imprinted genes on the X chromosome affecting brain development, neuropsychological development will depend on the parental origin of the single X chromosome. We contrasted verbal and visuospatial memory in females with a single paternal X chromosome (45,X(p)) and those with a single maternal X (45,X(m)). Neither group showed any impairment on immediate story recall; if anything, performance was above control levels. Groups did not differ on a measure of delayed recall. However, when delayed recall was considered after adjusting for level of immediate recall, 45,X(m) females showed enhanced verbal forgetting relative to controls over a delay. On the Rey figure, both groups were poor at copying the figure, but, after adjusting scores for initial copy score and strategy, only the 45,X(p) females showed disproportionate forgetting relative to controls. We propose there may be one or more imprinted genes on the X chromosome that affect the development of lateralised brain regions important for memory function.


Immediate effect of slow pace bhastrika pranayama on blood pressure and heart rate.
The objective of this study was to evaluate the immediate effect of slow pace bhastrika pranayama (respiratory rate 6/min) for 5 minutes on heart rate and blood pressure and the effect of the same breathing exercise for the same duration of time (5 minutes) following oral intake of hyoscine-N-butylbromide (Buscopan), a parasympathetic blocker drug. Heart rate and blood pressure of volunteers (n = 39, age = 25-40 years) was recorded following standard procedure. First, subjects had to sit comfortably in an easy and steady posture (sukhasana) on a fairly soft seat placed on the floor keeping head, neck, and trunk erect, eyes closed, and the other muscles reasonably loose. The subject is directed to inhale through both nostrils slowly up to the maximum for about 4 seconds and then exhale slowly up to the maximum through both nostrils for about 6 seconds. The breathing must not be abdominal. These steps complete one cycle of slow pace bhastrika pranayama (respiratory rate 6/min). During the practice the subject is asked not to think much about the inhalation and exhalation time, but rather was requested to imagine the open blue sky. The pranayama was conducted in a cool, well-ventilated room (18-20 degrees C). After 5 minutes of this breathing practice, the blood pressure and heart rate again were recorded in the aforesaid manner using the same instrument. The other group (n = 10) took part in another study where their blood pressure and heart rate were recorded following half an hour of oral intake of hyoscine-N-butylbromide 20 mg. Then they practiced the breathing exercise as stated above, and the abovementioned parameters were recorded again to study the effect of parasympathetic blockade on the same pranayama. It was noted that after slow bhastrika pranayamic breathing (respiratory rate 6/min) for 5 minutes, both the systolic and diastolic blood pressure decreased significantly with a slight fall in heart rate. No significant alteration in both blood pressure and heart rate was observed in volunteers who performed the same breathing exercise for the same duration following oral intake of hyoscine-N-butylbromide. Pranayama increases frequency and duration of inhibitory neural impulses by activating pulmonary stretch receptors during above tidal volume inhalation as in Hering Bruer reflex, which bring about withdrawal of sympathetic tone in the skeletal muscle blood vessels, leading to widespread vasodilatation, thus causing decrease in peripheral resistance and thus decreasing the diastolic blood pressure. After hyoscine-N-butylbromide, the parasympathetic blocker, it was observed that blood pressure was not decreased significantly as a result of pranayama, as it was observed when no drug was administered. Vagal cardiac and pulmonary mechanisms are linked, and improvement in one vagal limb might spill over into the other. Baroreceptor sensitivity can be enhanced significantly by slow breathing (supported by a small reduction in the heart rate observed during slow breathing and by reduction in both systolic and diastolic pressure). Slow pace bhastrika pranayama (respiratory rate 6/min) exercise thus shows a strong tendency to improving the autonomic nervous system through enhanced activation of the parasympathetic system.


[Urinary tract infection in pregnancy].
Several factors cause urinary tract infection (UTI) to be a relevant complication of the gestational period, aggravating both the maternal and perinatal prognosis. For many years, pregnancy has been considered to be a factor predisposing to all forms of UTI. Today, it is known that pregnancy, as an isolated event, is not responsible for a higher incidence of UTI, but that the anatomical and physiological changes imposed on the urinary tract by pregnancy predispose women with asymptomatic bacteriuria (AB) to become pregnant women with symptomatic UTI. AB affects 2 to 10% of all pregnant women and approximately 30% of these will develop pyelonephritis if not properly treated. However, a difficult-to-understand resistance against the identification of AB during this period is observed among prenatalists. The diagnosis of UTI is microbiological and it is based on two urine cultures presenting more than 10(5) colonies/mL urine of the same germ. Treatment is facilitated by the fact that it is based on an antibiogram, with no scientific foundation for the notion that a pre-established therapeutic scheme is an adequate measure. For the treatment of pyelonephritis, it is not possible to wait for the result of culture and previous knowledge of the resistance profile of the antibacterial agents available for the treatment of pregnant women would be the best measure. Another important variable is the use of an intravenous bactericidal antibiotic during the acute phase, with the possibility of oral administration at home after clinical improvement of the patient. At our hospital, the drug that best satisfies all of these requirements is cefuroxime, administered for 10-14 days. Third-generation cephalosporins do not exist in the oral form, all of them involving the inconvenience of parenteral administration. In view of their side effects, aminoglycosides are considered to be inadequate for administration to pregnant women. The inconsistent insinuation of contraindication of monofluorinated quinolones, if there is an indication, norfloxacin is believed to be a good alternative to cefuroxime. In cases in which UTI prophylaxis is indicated, chemotherapeutic agents are preferred, among them nitrofurantoin, with care taken to avoid its use at the end of pregnancy due to the risk of kernicterus for the neonate.


Use of Bioluminescence Markers To Detect Pseudomonas spp. in the Rhizosphere.
The use of bioluminescence as a sensitive marker for detection of Pseudomonas spp. in the rhizosphere was investigated. Continuous expression of the luxCDABE genes, required for bioluminescence, was not detectable in the rhizosphere. However, when either a naphthalene-inducible luxCDABE construct or a constitutive luxAB construct (coding only for the luciferase) was introduced into the Pseudomonas cells, light emission could be initiated just prior to measurement by the addition of naphthalene or the substrate for luciferase, n-decyl aldehyde, respectively. These Pseudomonas cells could successfully be detected in the rhizosphere by using autophotography or optical fiber light measurement techniques. Detection required the presence of 10 to 10 CFU/cm of root, showing that the bioluminescence technique is at least 1,000-fold more sensitive than beta-galactosidase-based systems.


The structural variation landscape in 492 Atlantic salmon genomes.
Structural variants (SVs) are a major source of genetic and phenotypic variation, but remain challenging to accurately type and are hence poorly characterized in most species. We present an approach for reliable SV discovery in non-model species using whole genome sequencing and report 15,483 high-confidence SVs in 492 Atlantic salmon (Salmo salar L.) sampled from a broad phylogeographic distribution. These SVs recover population genetic structure with high resolution, include an active DNA transposon, widely affect functional features, and overlap more duplicated genes retained from an ancestral salmonid autotetraploidization event than expected. Changes in SV allele frequency between wild and farmed fish indicate polygenic selection on behavioural traits during domestication, targeting brain-expressed synaptic networks linked to neurological disorders in humans. This study offers novel insights into the role of SVs in genome evolution and the genetic architecture of domestication traits, along with resources supporting reliable SV discovery in non-model species.


Good clinical practice in resource-limited settings: translating theory into practice.
A Good Clinical Practices (GCPs) course, based on the combination of theoretical modules with a practical training in real-life conditions, was held in 2010 in Burkina Faso. It was attended by 15 trainees from nine African, Asian, and Latin American countries. There were some discrepancies between the average good results at the end of the theoretical phase and the GCP application during the first days of the practical phase, underlying the difficulties of translating theoretical knowledge into good practices. Most of the findings were not unexpected and reflected the challenges commonly faced by clinical investigators in resource-poor contexts (i.e., the high workload at peripheral health facilities, the need to conciliate routine clinical activities with clinical research, and the risk of creating a double standard among patients attending the same health facility [free care for recruited patients versus user fees for non-recruited patients with the same medical condition]). Even if limited in number and time, these observations suggest that a theoretical training alone may not be sufficient to prepare trainees for the challenges of medical research in real-life settings. Conversely, when a practical phase immediately follows a theoretical one, trainees can immediately experience what the research methodology implicates in terms of work organization and relationship with recruited and non-recruited patients. This initial experience shows the complexity of translating GCP into practice and suggests the need to rethink the current conception of GCP training.


Epidemiology and clinical characteristics of autoimmune hepatitis in the Netherlands.
Epidemiological data on autoimmune hepatitis (AIH) are scarce. In this study, we determined the clinical and epidemiological characteristics of AIH patients in the Netherlands (16.7 million inhabitants). Clinical characteristics were collected from 1313 AIH patients (78% females) from 31 centers, including all eight academic centers in the Netherlands. Additional data on ethnicity, family history and symptoms were obtained by the use of a questionnaire. The prevalence of AIH was 18.3 (95% confidential interval [CI]: 17.3-19.4) per 100,000 with an annual incidence of 1.1 (95% CI: 0.5-2) in adults. An incidence peak was found in middle-aged women. At diagnosis, 56% of patients had fibrosis and 12% cirrhosis in liver biopsy. Overall, 1% of patients developed HCC and 3% of patients underwent liver transplantation. Overlap with primary biliary cirrhosis and primary sclerosing cholangitis was found in 9% and 6%, respectively. The clinical course did not differ between Caucasian and non-Caucasian patients. Other autoimmune diseases were found in 26% of patients. Half of the patients reported persistent AIH-related symptoms despite treatment with a median treatment period of 8 years (range 1-44 years). Familial occurrence was reported in three cases. This is the largest epidemiological study of AIH in a geographically defined region and demonstrates that the prevalence of AIH in the Netherlands is uncommon. Although familial occurrence of AIH is extremely rare, our twin data may point towards a genetic predisposition. The high percentage of patients with cirrhosis or fibrosis at diagnosis urges the need of more awareness for AIH.


Effects of a carbohydrate-restricted diet with and without supplemental soluble fiber on plasma low-density lipoprotein cholesterol and other clinical markers of cardiovascular risk.
Carbohydrate-restricted diets (CRDs) promote weight loss, reductions in plasma triacylglycerol (TAG) levels, and increases in high-density lipoprotein cholesterol (HDL-C) levels but may cause undesirable low-density lipoprotein cholesterol (LDL-C) responses in some people. The objective of the present study was to determine the effect of adding soluble fiber to a CRD on plasma LDL-C and other traditionally measured markers of cardiovascular disease. Using a parallel-arm, double-blind, placebo-controlled design, 30 overweight and obese men (body mass index, 25-35 kg/m(2)) were randomly assigned to supplement a CRD with soluble fiber (Konjac-mannan, 3g/d) (n = 15) or placebo (n = 15). Plasma lipids, anthropometrics, body composition, blood pressure, and nutrient intake were evaluated at baseline and at 6 and 12 weeks. Compliance was excellent as assessed by 7-day weighed dietary records and ketonuria. Both groups experienced decreases in (P < .01) body weight, percent body fat, systolic blood pressure, waist circumference, and plasma glucose levels. After 12 weeks, HDL-C and TAG improved significantly in the fiber (10% and -34%) and placebo (14%, -43%) groups. LDL-C decreased by 17.6% (P < .01) at week 6 and 14.1% (P < .01) at week 12 in the fiber group. Conversely, LDL-C reductions were significant in the placebo group only after 12 weeks (-6.0%, P < .05). We conclude that although clearly effective at lowering LDL-C, adding soluble fiber to a CRD during active and significant weight loss provides no additional benefits to the diet alone. Furthermore, a CRD led to clinically important positive alterations in cardiovascular disease risk factors.


Dickkopf-3, a tissue-derived modulator of local T-cell responses.
The adaptive immune system protects organisms from harmful environmental insults. In parallel, regulatory mechanisms control immune responses in order to assure preservation of organ integrity. Yet, molecules involved in the control of T-cell responses in peripheral tissues are poorly characterized. Here, we investigated the function of Dickkopf-3 in the modulation of local T-cell reactivity. Dkk3 is a secreted, mainly tissue-derived protein with highest expression in organs considered as immune-privileged such as the eye, embryo, placenta, and brain. While T-cell development and activation status in naïve Dkk3-deficient mice was comparable to littermate controls, we found that Dkk3 contributes to the immunosuppressive microenvironment that protects transplanted, class-I mismatched embryoid bodies from T-cell-mediated rejection. Moreover, genetic deletion or antibody-mediated neutralization of Dkk3 led to an exacerbated experimental autoimmune encephalomyelitis (EAE). This phenotype was accompanied by a change of T-cell polarization displayed by an increase of IFNγ-producing T cells within the central nervous system. In the wild-type situation, Dkk3 expression in the brain was up-regulated during the course of EAE in an IFNγ-dependent manner. In turn, Dkk3 decreased IFNγ activity and served as part of a negative feedback mechanism. Thus, our findings suggest that Dkk3 functions as a tissue-derived modulator of local CD4(+) and CD8(+) T-cell responses.


A systematic review of economic analyses of telehealth services using real time video communication.
Telehealth is the delivery of health care at a distance, using information and communication technology. The major rationales for its introduction have been to decrease costs, improve efficiency and increase access in health care delivery. This systematic review assesses the economic value of one type of telehealth delivery--synchronous or real time video communication--rather than examining a heterogeneous range of delivery modes as has been the case with previous reviews in this area. A systematic search was undertaken for economic analyses of the clinical use of telehealth, ending in June 2009. Studies with patient outcome data and a non-telehealth comparator were included. Cost analyses, non-comparative studies and those where patient satisfaction was the only health outcome were excluded. 36 articles met the inclusion criteria. 22(61%) of the studies found telehealth to be less costly than the non-telehealth alternative, 11(31%) found greater costs and 3 (9%) gave the same or mixed results. 23 of the studies took the perspective of the health services, 12 were societal, and one was from the patient perspective. In three studies of telehealth to rural areas, the health services paid more for telehealth, but due to savings in patient travel, the societal perspective demonstrated cost savings. In regard to health outcomes, 12 (33%) of studies found improved health outcomes, 21 (58%) found outcomes were not significantly different, 2(6%) found that telehealth was less effective, and 1 (3%) found outcomes differed according to patient group. The organisational model of care was more important in determining the value of the service than the clinical discipline, the type of technology, or the date of the study. Delivery of health services by real time video communication was cost-effective for home care and access to on-call hospital specialists, showed mixed results for rural service delivery, and was not cost-effective for local delivery of services between hospitals and primary care.


Protein expression profiling of the shrimp cellular response to white spot syndrome virus infection.
To better understand the pathogenesis of white spot syndrome virus (WSSV) and to determine which cell pathways might be affected after WSSV infection, two-dimensional gel electrophoresis (2-DE) was used to produce protein expression profiles from samples taken at 48 h post-infection (hpi) from the stomachs of Litopenaeus vannamei (also called Penaeus vannamei) that were either specific pathogen free or else infected with WSSV. Seventy-five protein spots that consistently showed either a marked change (>50%) in accumulated levels or else were highly expressed throughout the course of WSSV infection were selected for further study. After in-gel trypsin digestion followed by LC-nanoESI-MS/MS, bioinformatics databases were searched for matches. A total of 53 proteins were identified, with functions that included energy production, calcium homeostasis, nucleic acid synthesis, signaling/communication, oxygen carrier/transportation, and SUMO-related modification. 2-DE results were shown to be consistent with relative EST database data from a previously developed EST database of two Penaeus monodon cDNA libraries. For seven selected genes, 2-DE and EST data were also compared with transcriptional time-course RT-PCR data. This study is the first global analysis of differentially expressed proteins in WSSV-infected shrimp, and in addition to increasing our understanding of the molecular pathogenesis of this virus-associated shrimp disease, the results presented here should be useful both for identifying potential biomarkers and for developing antiviral measures.


Urinary liver-type fatty acid-binding protein is associated with subendocardial viability ratio in middle- and older-aged adults.
To identify one of the physiological underlying mechanisms of the predictive effects of urinary liver-type fatty acid-binding protein (L-FABP) for the onset of cardiovascular disease (CVD), we investigated the relationship between urinary L-FABP levels and subendocardial viability ratio (SEVR), a marker of myocardial perfusion, in middle- and older-aged adults. This was a cross-sectional study of 249 middle- and older-aged adults (aged 46-83 years). We measured urinary L-FABP levels and central hemodynamic parameters, including SEVR, calculated by pulse wave analysis using an applanation tonometry. In the participants stratified in accordance with the tertiles of urinary L-FABP levels, SEVR decreased in a stepwise fashion with increasing tertiles (p < 0.001). Furthermore, this association remains significant after the consideration of various confounders. On the contrary, urinary albumin levels were not independently related with SEVR. Our results demonstrated that urinary L-FABP levels were independently associated with the SEVR of the middle- and older-aged adults. This result suggests that the increase in urinary L-FABP levels even within the normal range might be related to the decrease in myocardial perfusion (SEVR).


Quantitative genetic and translocation experiments reveal genotype-by-environment effects on juvenile life-history traits in two populations of Chinook salmon (Oncorhynchus tshawytscha).
Understanding the genetic architecture of phenotypic plasticity is required to assess how populations might respond to heterogeneous or changing environments. Although several studies have examined population-level patterns in environmental heterogeneity and plasticity, few studies have examined individual-level variation in plasticity. Here, we use the North Carolina II breeding design and translocation experiments between two populations of Chinook salmon to detail the genetic architecture and plasticity of offspring survival and growth. We followed the survival of 50,800 offspring through the larval stage and used parentage analysis to examine survival and growth through freshwater rearing. In one population, we found that additive genetic, nonadditive genetic and maternal effects explained 25%, 34% and 55% of the variance in larvae survival, respectively. In the second population, these effects explained 0%, 24% and 61% of the variance in larvae survival. In contrast, fry survival was regulated primarily by additive genetic effects, which indicates a shift from maternal to genetic effects as development proceeds. Fry growth also showed strong additive genetic effects. Translocations between populations revealed that offspring survival and growth varied between environments, the degree of which differed among families. These results indicate genetic differences among individuals in their degree of plasticity and consequently their ability to respond to environmental variation.


Prognostic value of acquired uniparental disomy (aUPD) in primary breast cancer.
Many studies have examined DNA copy number changes or gene expression profiling and their association with clinical outcomes in breast cancer. However, until now no study has investigated whether acquired uniparental disomy (aUPD), in which both chromosomes in a pair are derived from the same parent, may have an association with clinical outcome including initiation and recurrence of breast cancer. In this study, we used high-density SNP and expression microarrays data from primary tumors of 313 lymph node-negative breast cancer patients who had not received adjuvant systemic therapy to evaluate the association of aUPD with metastasis-free survival (MFS) and overall survival (OS). In 55.9% (175/313) of the tumors, we defined aUPD, which was most frequent in the regions at chr17q (30.3%) and chr13q (19.4%). In Cox univariate regression analysis including all patients, aUPD at four regions at chr17q, ranging in size from 2.9 to 4.0 Mb, were associated with a poor OS. Only aUPD at one region, region B, on chr17q was associated with a poor MFS. Similarly, aUPD at two regions, A and B, on chr13q, with sizes of 3.5 and 3.1 Mb, were associated with a poor OS, but not with MFS. In ER-subgroup analyses, regions B and D at 17q were associated with poor MFS and OS in ER-negative patients. Various differentially expressed genes within the identified aUPD regions at chr17q were associated with MFS and OS in all patients (PPM1D, C17orf71, and TRIM37) and/or in the ER-negative patients (PPM1D, PPM1E, and SLCA3R1). We thus conclude that aUPD is a frequent event in breast cancer and that aUPD at specific regions in the genome has implications in this disease.