Matrix metalloproteinases are involved in mechanical stretch-induced activation of skeletal muscle satellite cells.
When skeletal muscle is stretched or injured, myogenic satellite cells are activated to enter the cell cycle. This process depends on nitric oxide (NO) production, release of hepatocyte growth factor (HGF) from the extracellular matrix, and presentation of HGF to the c-met receptor. Experiments reported herein provide new evidence that matrix metalloproteinases (MMPs) are involved in the NO-dependent release of HGF in vitro. When rat satellite cells were treated with 10 ng/ml recombinant tissue inhibitor-1 of MMPs (TIMP-1) and subjected to treatments that induce activation in vitro, i.e., sodium nitroprusside (SNP) of an NO donor or mechanical cyclic stretch, the activation response was inhibited. In addition, conditioned medium generated by cultures treated with TIMP-1 plus SNP or mechanical stretch failed to activate cultured satellite cells and did not contain HGF. Moreover, NO(x) assay demonstrated that TIMP-1 does not impair NO synthase activity of stretched satellite cell cultures. Therefore, results from these experiments provide strong evidence that MMPs mediate HGF release from the matrix and that this step in the pathway is downstream from NO synthesis.


Intestinal transplantation: indications, results and strategy.
The term 'intestinal failure' is now often used to describe gastrointestinal function insufficient to satisfy body nutrient and fluid requirements. The first recognized condition of intestinal failure was short bowel syndrome. Severe motility disorders such as chronic intestinal pseudo-obstruction syndrome in children as well as congenital intractable intestinal mucosa disorders are also forms of intestinal failure, because no curative treatment for these diseases is yet available. Parenteral nutrition and home parenteral nutrition remain the mainstay of therapy for intestinal failure, whether it is partial or total, provisional or permanent. However, some patients develop complications while receiving standard therapy for intestinal failure and are considered for intestinal transplantation. Indeed, recent advances in immunosuppressive treatment and the better monitoring and control of acute rejection have brought intestinal transplantation into the realm of standard treatment for intestinal failure. Although it has been used in humans for the past two decades, this procedure has had a slow learning curve. According to the current results, this challenging procedure may be performed in children or adults, only under certain conditions.


Effects of emergence time and early social rearing environment on behaviour of Atlantic salmon: consequences for juvenile fitness and smolt migration.
Consistent individual differences in behaviour have been well documented in a variety of animal taxa, but surprisingly little is known about the fitness and life-history consequences of such individual variation. In wild salmonids, the timing of fry emergence from gravel spawning nests has been suggested to be coupled with individual behavioural traits. Here, we further investigate the link between timing of spawning nest emergence and behaviour of Atlantic salmon (Salmo salar), test effects of social rearing environment on behavioural traits in fish with different emergence times, and assess whether behavioural traits measured in the laboratory predict growth, survival, and migration status in the wild. Atlantic salmon fry were sorted with respect to emergence time from artificial spawning nest into three groups: early, intermediate, and late. These emergence groups were hatchery-reared separately or in co-culture for four months to test effects of social rearing environment on behavioural traits. Twenty fish from each of the six treatment groups were then subjected to three individual-based behavioural tests: basal locomotor activity, boldness, and escape response. Following behavioural characterization, the fish were released into a near-natural experimental stream. Results showed differences in escape behaviour between emergence groups in a net restraining test, but the social rearing environment did not affect individual behavioural expression. Emergence time and social environment had no significant effects on survival, growth, and migration status in the stream, although migration propensity was 1.4 to 1.9 times higher for early emerging individuals that were reared separately. In addition, despite individuals showing considerable variation in behaviour across treatment groups, this was not translated into differences in growth, survival, and migration status. Hence, our study adds to the view that fitness (i.e., growth and survival) and life-history predictions from laboratory measures of behaviour should be made with caution and ideally tested in nature.


Family Characteristics Associated With Child Maltreatment Across the Deployment Cycle of U.S. Army Soldiers.
Soldier deployment can create a stressful environment for U.S. Army families with young children. Prior research has identified elevated rates of child maltreatment in the 6 months immediately following a soldier's return home from deployment. In this study, we longitudinally examine how other child- and family-level characteristics influence the relationship of deployment to risk for maltreatment of dependent children of U.S. Army soldiers. We conducted a person-time analysis of substantiated reports and medical diagnoses of maltreatment among the 73,404 children of 56,087 U.S. Army soldiers with a single deployment between 2001 and 2007. Cox proportional hazard models estimated hazard rates of maltreatment across deployment periods and simultaneously considered main effects for other child- and family-level characteristics across periods. In adjusted models, maltreatment hazard was highest in the 6 months following deployment (hazard ratio [HR] = 1.63, p < 0.001). Children born prematurely or with early special needs independently had an increased risk for maltreatment across all periods (HR = 2.02, p < 0.001), as well as those children whose soldier-parent had been previously diagnosed with a mental illness (HR = 1.68, p < 0.001). In models testing for effect modification, during the 6 months before deployment, children of female soldiers (HR = 2.22, p = 0.006) as well as children of soldiers with a mental health diagnosis (HR = 2.78, p = 0.001) were more likely to experience maltreatment, exceeding the risk at all other periods. Infants and children are at increased risk for maltreatment in the 6 months following a parent's deployment, even after accounting for other known family- and child-level risk factors. However, the risk does not appear to be the same for all soldiers and their families in relation to deployment, particularly for female soldiers and those who had previously diagnosed mental health issues, for whom the risk appears most elevated before deployment. Accounting for the unique needs of high-risk families at different stages of a soldier's deployment cycle may allow the U.S. Army to better direct resources that prevent and address child maltreatment.


Cytogenetic and molecular cytogenetic findings in dedifferentiated liposarcoma with neural-like whorling pattern and metaplastic bone formation.
Dedifferentiated liposarcoma is a subtype of liposarcoma characterized by juxtaposition of well-differentiated liposarcoma with a nonadipocytic sarcoma. A peculiar form of dedifferentiated liposarcoma has been described, characterized by a nonlipogenic component with a neural-like whorling pattern of growth and metaplastic bone formation. We report the cytogenetic and molecular genetic findings of this peculiar form of dedifferentiation in a retroperitoneal tumor found in a 58-year-old female. The neoplasm had typical histologic findings and a complex karyotype characterized by several numeric and structural chromosomal abnormalities, including the presence of ring and giant rod chromosomes. Molecular cytogenetic studies found high levels of amplification of the MDM2 oncogene, consistent with the amplification of the 12q14 chromosome band, a cytogenetic abnormality commonly found in these tumors. These findings indicate that, despite its unique and peculiar morphologic features, this unusual type of dedifferentiated liposarcoma shares many of the cytogenetic features and molecular genetic abnormalities found in other forms of dedifferentiation. The specific cytogenetics and molecular determinants of these peculiar morphologic findings, however, remain unknown.


Tight focusing of vortex beams in presence of primary astigmatism.
The tight focusing of vortex carrying beams is studied using the Debye-Wolf diffraction integral in the presence of primary astigmatism. The roles of topological charge, polarization distribution of the input beam, and handedness of the beam polarization are investigated in the intensity distribution of the focal plane of a high-numerical-aperture lens. The effect of tight focusing in the presence of astigmatism on the dark core of the azimuthally polarized beam is also investigated and compared with the dark core of a circularly polarized vortex beam. The effect of an aberration has been discussed in the context of the fluorescent spot size in the focal plane of a stimulated emission depletion microscope for two different polarization setups.


High resolution melting analysis for the differentiation of Mycobacterium species.
A quantitative real-time PCR (qPCR) followed by high resolution melting (HRM) analysis was developed for the differentiation of Mycobacterium species. Rapid differentiation of Mycobacterium species is necessary for the effective diagnosis and management of tuberculosis. In this study, the 16S rRNA gene was tested as the target since this has been identified as a suitable target for the identification of mycobacteria species. During the temperature gradient and primer optimization process, the melting peak (Tm) analysis was determined at a concentration of 50 ng DNA template and 0.3, 0.4 and 0.5 µM primer. The qPCR assay for the detection of other mycobacterial species was done at the Tm and primer concentration of 62 °C and 0.4 µM, respectively. The HRM analysis generated cluster patterns that were specific and sensitive to distinguished small sequence differences of the Mycobacterium species. This study suggests that the 16S rRNA-based real-time PCR followed by HRM analysis produced unique cluster patterns for species of Mycobacterium and could differentiate the closely related mycobacteria species.


Characterization of cell envelope proteins of Staphylococcus epidermidis cultured in human peritoneal dialysate.
The cell envelope protein profiles of Staphylococcus epidermidis cultured in used human peritoneal dialysate (HPD) differed markedly from those of cells cultured in nutrient broth. Compared with broth-grown cells, many cell wall proteins were repressed in HPD, although three proteins of 42, 48, and 54 kDa predominated and an iron-repressible 130-kDa protein was induced. Growth in HPD also resulted in expression of two cell membrane proteins of 32 and 36 kDa which were iron repressible. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis using monospecific polyclonal antisera raised against the 32- and 36-kDa proteins revealed considerable antigenic and molecular mass homology among 12 S. epidermidis isolates from patients with continuous ambulatory peritoneal dialysis-related peritonitis. The 32-kDa antiserum also cross-reacted with a 32-kDa S. aureus cell membrane protein. Immunoblots of S. epidermidis cell walls and membranes were also probed with normal human serum and serum and HPD from continuous ambulatory peritoneal dialysis patients. While the cell wall proteins of S. epidermidis appeared to be relatively poorly immunogenic, the 32- and 36-kDa membrane proteins reacted strongly with antibodies present in each of the body fluids evaluated. These results suggest that the highly conserved 32- and 36-kDa iron-repressible proteins are expressed during growth in vivo and may be involved in iron transport, since all 12 S. epidermidis strains examined also produced iron chelators.


Pulmonary hypertension and fenfluramine.
Pulmonary hypertension developed in two women who had been taking fenfluramine for over eight months for weight reduction. On withdrawing the drug symptoms and electrocardiographic evidence of pulmonary hypertension disappeared in both cases. In one patient, however, the evidence recurred after rechallenge with fenfluramine.These findings are strong evidence that fenfluramine may cause pulmonary hypertension. Hence any patient taking the drug should report immediately any deterioration in exercise tolerance.


Pretransfusion Comparison of Dialyser-Based Hemoconcentrator With Cell Saver System for Perioperative Cell Salvage.
Cell Saver system is the method of choice for red blood cell salvage from the surgical field; however, cost is a limiting factor. We at our institute have devised a cost-effective version of dialyser-based autotransfusion system. We performed pretransfusion comparison of our autotransfusion system with conventional cell saver system. A prospective randomized observational study was performed in 104 consecutive patients with coronary artery disease undergoing by off-pump coronary artery bypass grafting. Patients were divided into two groups. In the dialyser group (53 patients), blood from surgical field was salvaged by our dialyser-based system. In the cell saver group (51 patients), blood was salvaged by cell saver. In both groups, 20-mL sample from the salvaged blood was analyzed for hemoglobin, platelets, protein, albumin, free hemoglobin, osmotic fragility, and peripheral blood smear examination. Total hemoglobin salvaged was comparable in both groups (85% vs 76%). On peripheral smear, red blood cells were swollen, but morphology was preserved. Moreover, normal osmotic fragility suggested absence of any lethal damage to red blood cells in either group. Dialyser-based system was more efficient in salvaging platelets (42.9% vs 6%), proteins (79.2% vs 0%), and albumin (65% vs 0%). Total free hemoglobin was three times more in dialyser group but was well below recommended limits. Dialyser-based system is economical, is equally efficacious in salvaging red blood cells, is more effective in salvaging platelets and proteins, and does not contain significant amount of free hemoglobin. Therefore, this salvaged blood can be safely transfused.


Linear vector operations in coherent optical data processing systems.
The generation of a two-dimensional linear vector space in a coherent optical data processing system and its specific application to the gradient operator is discussed using theoretical and experimental results. The vector space is created by superposing the outputs of two Fourier optical systems having light of mutually orthogonal polarizations. As a result, the total amplitude of the signal in the output plane is a vector sum of the signals from the systems. If each one of the systems performs one of the partial derivative operations of the transverse gradient, and if the inputs to both systems are identical, then the output is the vector sum of the partial derivatives or the transverse gradient operation. The experimental program is heavily oriented toward the realization of the optimum approximation to the jomega(x)î and jomega(y)j; filters, rather than using various binary-type filters. Problems with film and lens noise are also discussed.


Cytotoxic effects of benzbromarone and its 1'-hydroxy metabolite in human hepatocarcinoma FLC4 cells cultured on micro-space cell culture plates.
Treatment with benzbromarone (BBR), a potent uricosuric drug, can be associated with liver injury. Recently, we reported that culture of human hepatocellular carcinoma FLC-4 cells on micro-space cell culture plates could increase the functional expression of drug-metabolizing enzymes including CYP3A4 and CYP2C9, which are involved in 1'-hydroxylation and 6-hydroxylation of BBR, respectively. Therefore, we examined whether BBR and its two metabolites (1'-hydroxy BBR and 6-hydroxy BBR) have cytotoxic effects in FLC4 cells cultured on micro-space cell culture plates. The present study showed that BBR and 1'-hydroxy BBR, but not 6-hydroxy BBR, have cytotoxic effects in cells cultured on micro-space cell culture plates. BBR-induced cytotoxicity was decreased by CYP3A inhibitors (itraconazole and ketoconazole), an Nrf2 activator (tert-butylhydroquinone) and a GSH precursor (N-acetyl-L-cystein). In contrast, BBR-induced cytotoxicity was increased by a GSH biosynthesis inhibitor (buthionine sulfoximine) and an inhibitor of NAD(P)H quinone oxidoreductase 1 (dicoumarol). These results suggested that metabolic activation of 1'-hydroxy BBR via CYP3A, formation of quinone metabolites and the decrease in GSH levels were involved in the BBR-induced cytotoxicity observed in FLC4 cells cultured on micro-space cell culture plates.


Influence of foot position on the measurement of first metatarsal axial rotation using the first metatarsal axial radiographs.
The purpose of this study was to clarify 1) the measurement error of the pronation angle using the first metatarsal axial radiograph with the pronation angle along the longitudinal axis of the first metatarsal as the reference standard, 2) the influence of variability in the foot position on the measurement error, and 3) the intra- and interrater reliability of pronation angle measurement using digitally reconstructed radiographs. Digitally reconstructed radiographs of the first metatarsal were generated from the computed tomography images of 10 feet without hallux valgus (non-HV group) and 10 feet with hallux valgus (HV group). In total, 135 images were created at different degrees of supination, plantarflexion, and adduction from each foot to simulate the first metatarsal axial view. Then, the pronation angle of the first metatarsal was measured. The measurement error was determined using the mean error and 95% limits of agreement. Simple linear regression analysis was used to test the correlations of the measurement error with pronation, plantarflexion and adduction angles. The intra- and interrater reliability of measurement was assessed using the intraclass correlation coefficient and minimum detectable change values. The mean measurement errors were 0.1° for both the non-HV and HV groups. There was no significant correlation of the measurement error with pronation, plantarflexion or adduction angles for both groups. Additionally, the intraclass correlation coefficients for the intra- and interrater reliability were more than 0.9 in both the non-HV and HV groups with the minimum detectable change values ranging from 0.7° to 1.4°. The measurement error of first metatarsal pronation using the axial view was clinically acceptable. The measurements were not influenced by the variability in foot position while obtaining the radiograph. The first metatarsal axial view could be used to quantify the first metatarsal coronal rotation.


Sesamol ameliorates cyclophosphamide-induced hepatotoxicity by modulating oxidative stress and inflammatory mediators.
In current scenario of human health and diseases, drug-induced hepatic injury has been recognized as a serious and unresolved problem. Particularly, chemotherapeutic agents have been reported to induce organ toxicity. The aim of the present study is to evaluate organ toxicity and oxidative damage induced by cyclophosphamide (CP), a chemotherapeutic drug and its amelioration by sesamol, an antioxidant from sesame seeds. CP (150 mg/kg) is injected intraperitonially to experimental rats and from day 2 rats were orally treated with sesamol. Rats were sacrificed to evaluate non-enzymatic and enzymatic oxidative stress parameters in serum and tissue homogenates on day 8. Besides, liver function parameters and pro-inflammatory mediators were assessed. Histopathological studies of liver and kidney were also carried out. Elevated levels of endogenous reactive oxygen species, lipid peroxidation and decreased levels of glutathione, total thiols, along with the reduction in antioxidant enzymes including superoxide dismutase, catalase, glutathione-stransferase and glutathione peroxidase, were evident in CP-intoxicated animals. Pro-inflammatory mediators like tumor necrosis factor - α, interleukin (IL)-1β, IL-6 and cyclooxygenase-2 were also elevated. Moreover, the levels of liver function markers like serum alanine aminotransferase and aspartate aminotransferase were also altered. Histology of liver and kidney tissues further supported CP-induced organ damage. Altered parameters were significantly restored to normal by oral administration of sesamol (50 mg/kg) suggesting its antioxidative stress, anti-inflammatory and hepatoprotective abilities. The study clearly demonstrated the potentiality of sesamol against CPinduced organ toxicity and oxidative stress suggesting its applicability in treatment regime of cancer and other stress-associated disorders as a supportive/auxiliary therapy.


Simplified categorization of outdoor activities for male and female U.S. indoor workers-a feasibility study to improve assessment of ultraviolet radiation exposures in epidemiologic study questionnaires.
Skin cancer studies depend on questionnaires to estimate exposure to ultraviolet light and subsequent risk but are limited by recall bias. We investigate the feasibility of developing a short checklist of categories comprising outdoor activities that can improve sun exposure questionnaires for use in epidemiologic studies. We recruited 124 working and retired U.S. radiologic technologists (52% women). Each subject was instructed to complete a daily activity diary, listing main indoor and outdoor activities between 9:00 A.M. and 5:00 P.M. during a 7 day period. A total of 4,697 entries were associated with 1408 h (21.1%) of the total 6,944 h spent outdoors. We were able to classify the activities into seven main activity categories: driving, yard work, home-maintenance, walking or performing errands, water activities, other recreational or sports activities and leisure activities or relaxing outside. These activities accounted for more than 94% of time spent outdoors both for working and retired men and women. Our data document the feasibility and guidance for developing a short checklist of outdoor activities for use in epidemiologic questionnaires for estimating sunlight exposures of working and retired indoor workers.


Integration of autopatching with automated pipette and cell detection in vitro.
Patch clamp is the main technique for measuring electrical properties of individual cells. Since its discovery in 1976 by Neher and Sakmann, patch clamp has been instrumental in broadening our understanding of the fundamental properties of ion channels and synapses in neurons. The conventional patch-clamp method requires manual, precise positioning of a glass micropipette against the cell membrane of a visually identified target neuron. Subsequently, a tight "gigaseal" connection between the pipette and the cell membrane is established, and suction is applied to establish the whole cell patch configuration to perform electrophysiological recordings. This procedure is repeated manually for each individual cell, making it labor intensive and time consuming. In this article we describe the development of a new automatic patch-clamp system for brain slices, which integrates all steps of the patch-clamp process: image acquisition through a microscope, computer vision-based identification of a patch pipette and fluorescently labeled neurons, micromanipulator control, and automated patching. We validated our system in brain slices from wild-type and transgenic mice expressing channelrhodopsin 2 under the Thy1 promoter (line 18) or injected with a herpes simplex virus-expressing archaerhodopsin, ArchT. Our computer vision-based algorithm makes the fluorescent cell detection and targeting user independent. Compared with manual patching, our system is superior in both success rate and average trial duration. It provides more reliable trial-to-trial control of the patching process and improves reproducibility of experiments.


Proteasome enzymatic activities in plasma as risk stratification of patients with acute myeloid leukemia and advanced-stage myelodysplastic syndrome.
Cytogenetic abnormalities are currently the most important predictors of response and clinical outcome for patients with acute myeloid leukemia (AML) or advanced-stage myelodysplastic syndrome (MDS). Because clinical outcomes vary markedly within cytogenetic subgroups, additional biological markers are needed for risk stratification. We assessed the utility of measuring pretreatment proteasome chymotrypsin-like, caspase-like, and trypsin-like activities in plasma to predict response and survival of patients with AML (n = 174) or advanced-stage MDS (n = 52). All three enzymatic activities were significantly (P < 0.001) increased in the plasma of patients with AML and MDS compared with normal controls. Both chymotrypsin-like and caspase-like activities, but not trypsin-like activity, correlated with outcome. Chymotrypsin-like and caspase-like activities, but not trypsin-like activity, predicted response in univariate analysis (P = 0.002). However, only chymotrypsin-like activity was independent predictor of response from age grouping (<70 versus > or =70 years), cytogenetics, and blood urea nitrogen in multivariate analysis. Similarly, both chymotrypsin-like and caspase-like activities, but not trypsin-like activity, were predictors of overall survival in univariate analysis (P < 0.0001), but only chymotrypsin-like activity was independent of cytogenetics, age, performance status, blood urea nitrogen, and beta(2)-microglobulin in multivariate Cox regression models. Chymotrypsin-like activity was also a strong independent predictor of survival in patients with intermediate karyotype (n = 124). Measuring plasma chymotrypsin-like activity may provide a powerful biomarker for risk stratification in patients with AML and advanced-stage MDS, including those with normal karyotype.


Soluble CD157 in pleural effusions: a complementary tool for the diagnosis of malignant mesothelioma.
CD157/Bst1 glycoprotein is expressed in >85% of malignant pleural mesotheliomas and is a marker of enhanced tumor aggressiveness. In vitro, mesothelial cells (malignant and non-malignant) released CD157 in soluble form or as an exosomal protein. In vivo, sCD157 is released and can be measured in pleural effusions by ELISA. Significantly higher levels of effusion sCD157 were detected in patients with malignant pleural mesothelioma than in patients with non-mesothelioma tumors or with non-malignant conditions. In our patient cohort, the area under the receiver-operating characteristic curve for sCD157 that discriminated malignant pleural mesothelioma from all other causes of pleural effusion was 0.685, cut-off (determined by the Youden Index) = 23.66 ng/ml (62.3% sensitivity; 73.93% specificity). Using a cut-off that yielded 95.58% specificity, measurement of sCD157 in cytology-negative effusions increased sensitivity of malignant pleural mesothelioma diagnosis from 34.42% to 49.18%. Evaluation of soluble CD157 in pleural effusions provides a diagnostic aid in malignant mesothelioma. Soluble CD157 (sCD157) was detected biochemically in culture supernatants of malignant and non-malignant mesothelial cells, and in pleural effusions from various pathological conditions. An ELISA system was established to measure the concentration of sCD157 in fluids, and extended to analyze sCD157 in pleural effusions from a cohort of 295 patients.


Theoretical Investigation of the Radical-Radical Reaction of O((3)P) + C2H3 and Comparison with Gas-Phase Crossed-Beam Experiments.
Herein, we present an ab initio study of the prototypal radical-radical reactions of ground-state atomic oxygen [O((3)P)] with the vinyl (C2H3) radical using density functional theory and a complete basis set model. Two distinctive pathways on the lowest doublet potential energy surfaces (PESs) were predicted to be in competition: addition and abstraction. The barrierless addition of O((3)P) to the hydrocarbon radicals leads to energy-rich intermediate formation followed by subsequent isomerization and decomposition to yield various products: CH2CO (ketene) + H, CO + CH3, C2HOH (acetylenol) + H, (3,1)CCHOH + H, H2O + C2H, (3,1)CH2 + HCO, H2CO (formaldehyde) + CH, C2H2 (acetylene) + OH, and (3,1)CCH2 + OH. The competing but minor H-atom abstraction mechanisms produce C2H2 + OH and (1,3)CCH2 + OH. The optimized structures of the reactants, products, intermediates, and transition states and the reaction mechanisms were obtained on the lowest doublet PESs. The major pathway was predicted to be the formation of CH2CO + H through the low-barrier, single-step cleavages of the addition intermediates. The Levine-Bernstein prior method, statistical surprisal approach, and microcanonical Rice-Ramsperger-Kassel-Marcus theory were applied to deduce the energy distributions of H atoms and OH products and quantitative rate constants. On the basis of the statistical theory and the population analysis, the predicted energy distributions were compared to the kinetic energy release of H and the preferential population of the Π(A') component of OH products reported in recent gas-phase crossed-beam investigations (Park, M. J.; Jang, S. C.; Choi, J. H. J. Chem. Phys. 2012, 137, 204311), and their kinetic and dynamic characteristics were discussed.


A novel-1364A/C aquaporin 5 gene promoter polymorphism influences the responses to salt loading of the renin-angiotensin-aldosterone system and of blood pressure in young healthy men.
The family of aquaporin water channels contributes to water and salt homeostasis. AQP5 is a ubiquitously expressed exocrine-type water channel. Functional single nucleotide polymorphisms in AQP5 which alter gene transcription have not yet been described. We, therefore, sequenced the human AQP5 promoter to detect novel sequence variants which could impact upon AQP5 expression and contribute to the phenotypic variability of the renin-angiotensin-aldosterone system (RAAS). Sequencing of the whole AQP5 promoter revealed a novel-1364A/C polymorphism. Substitution of C for A was associated with increased transcription factor binding as tested by electrophoretic mobility shift assay, but significantly reduced transcriptional activation of the AQP5 gene by cAMP and serum. The C allele was associated with significantly decreased mRNA in human heart and with decreased protein expression in erythrocyte membranes. Finally, we associated AQP5 genotypes with the variability of the RAAS in two independent study cohorts. First, we studied the phenotypic variability of the RAAS in 103 young (26 +/- 3 years) healthy males under an increased dietary salt intake. Increasing salt intake decreased plasma angiotensin II by 25% in AC/CC genotypes but only by 2% in AA genotypes (P = 0.012), and it decreased serum aldosterone by 34% in subjects with AC/CC genotypes but only by 19% in the AA genotypes (P = 0.005). Both genotypes had increased blood pressure under salt diet (P < 0.01), which was significantly more pronounced in AA genotypes (P = 0.029). Second, we investigated associations with variables of the RAAS in 96 old patients (68 +/- 10 years) with coronary artery disease scheduled for coronary artery bypass grafting. Aldosterone serum concentrations were 2-fold (P < 0.001) and angiotensin II plasma concentrations were 4-fold higher in AA genotypes than in AC/CC genotypes while ADH plasma concentrations did not differ. A novel single nucleotide polymorphism in the AQP5 gene promoter alters AQP5 expression in different in vitro systems and cells, and is associated with alterations of variables of the RAAS both in young healthy males and in patients with coronary artery disease.


The Popeye domain containing 2 (popdc2) gene in zebrafish is required for heart and skeletal muscle development.
The Popeye domain containing (Popdc) genes encode a family of transmembrane proteins with an evolutionary conserved Popeye domain. These genes are abundantly expressed in striated muscle tissue, however their function is not well understood. In this study we have investigated the role of the popdc2 gene in zebrafish. Popdc2 transcripts were detected in the embryonic myocardium and transiently in the craniofacial and tail musculature. Morpholino oligonucleotide-mediated knockdown of popdc2 resulted in aberrant development of skeletal muscle and heart. Muscle segments in the trunk were irregularly shaped and craniofacial muscles were severely reduced or even missing. In the heart, pericardial edema was prevalent in the morphants and heart chambers were elongated and looping was abnormal. These pathologies in muscle and heart were alleviated after reducing the morpholino concentration. However the heart still was abnormal displaying cardiac arrhythmia at later stages of development. Optical recordings of cardiac contractility revealed irregular ventricular contractions with a 2:1, or 3:1 atrial/ventricular conduction ratio, which caused a significant reduction in heart frequency. Recordings of calcium transients with high spatiotemporal resolution using a transgenic calcium indicator line (Tg(cmlc2:gCaMP)(s878)) and SPIM microscopy confirmed the presence of a severe arrhythmia phenotype. Our results identify popdc2 as a gene important for striated muscle differentiation and cardiac morphogenesis. In addition it is required for the development of the cardiac conduction system.


The entorhinal cortex of the monkey: II. Cortical afferents.
The entorhinal cortex of the monkey is commonly viewed as the major link between the cerebral cortex and the other fields of the hippocampal formation. Until recently, however, little was known about the origins of the cortical projections to the entorhinal cortex, and most of the available information is still based on degeneration studies. We have carried out a systematic analysis of these connections by placing small injections of the retrograde tracer wheat germ agglutinin conjugated to horseradish peroxidase into each of the fields of the entorhinal cortex of the Macaca fascicularis monkey. Retrogradely labeled cells were observed in several areas of the frontal and temporal lobes, the insula, and the cingulate cortex. In the frontal lobe, the greatest number of labeled cells were observed in the orbital region and specifically in areas 13 and 13a: labeled cells were also seen in areas 14, 11, and 12. In the dorsolateral frontal cortex, labeled cells were observed mainly in the rostral half of area 46; occasionally cells were also seen in areas 9, 8, and 6. In the cingulate cortex, labeled cells were observed in area 25, area 32, and rostral levels of area 24; fewer cells were observed at caudal levels of area 24 or in area 23. The retrosplenial region (areas 30 and 29), including its caudal extension along the rostral calcarine sulcus and its ventral extension into the temporal lobe, contained numerous labeled cells. In the temporal lobe, retrogradely labeled cells were arranged in two rostrocaudally oriented bands. Rostral to the hippocampal formation, the first band encompassed the piriform and periamygdaloid cortices and areas 35 and 36; the labeling in area 36 was continuous to the temporal pole. At more caudal levels this band was located immediately lateral to the hippocampal formation and included areas 35 and 36 rostrally and areas TH and TF caudally. The second band was situated in the superior temporal gyrus where labeled cells were observed in several distinct cytoarchitectonic fields, including the parainsular cortex in the fundus of the inferior limiting sulcus. In the insula proper, retrogradely labeled cells were seen mainly in the rostral or agranular division; far fewer were observed in the dysgranular and granular insula. Whereas there is little available physiological information concerning many of the cortical regions that project to the entorhinal cortex, on anatomical grounds they may be generally characterized as polysensory associational regions.


Controversies in the management of distal radius fractures.
Controversies span the entire spectrum of management of distal radius fractures-fracture assessment, diagnosis, treatment, and evaluation of outcomes. The utility of multiple radiographic views described in the literature has not been validated. Likewise, the several classification systems that exist have yet to demonstrate substantial interobserver and intraobserver reliability. Nonsurgical controversies involve fracture reduction, use of anesthesia, type of fracture immobilization, and forearm position during healing. Surgical controversies include surgical indications, need for release of carpal tunnel, fracture fixation method, and the need for augmentation (ie, bone graft). Postoperatively, rehabilitation, medication, and physical therapy also remain highly controversial. The best outcome measure has yet to be established. A strong need remains for high-level, prospective studies to determine the most effective way to assess, diagnose, treat, and measure outcomes in patients with distal radius fractures.


Impaired cell fate through gain-of-function mutations in a chromatin reader.
Modifications of histone proteins have essential roles in normal development and human disease. Recognition of modified histones by 'reader' proteins is a key mechanism that mediates the function of histone modifications, but how the dysregulation of these readers might contribute to disease remains poorly understood. We previously identified the ENL protein as a reader of histone acetylation via its YEATS domain, linking it to the expression of cancer-driving genes in acute leukaemia1. Recurrent hotspot mutations have been found in the ENL YEATS domain in Wilms tumour2,3, the most common type of paediatric kidney cancer. Here we show, using human and mouse cells, that these mutations impair cell-fate regulation by conferring gain-of-function in chromatin recruitment and transcriptional control. ENL mutants induce gene-expression changes that favour a premalignant cell fate, and, in an assay for nephrogenesis using murine cells, result in undifferentiated structures resembling those observed in human Wilms tumour. Mechanistically, although bound to largely similar genomic loci as the wild-type protein, ENL mutants exhibit increased occupancy at a subset of targets, leading to a marked increase in the recruitment and activity of transcription elongation machinery that enforces active transcription from target loci. Furthermore, ectopically expressed ENL mutants exhibit greater self-association and form discrete and dynamic nuclear puncta that are characteristic of biomolecular hubs consisting of local high concentrations of regulatory factors. Such mutation-driven ENL self-association is functionally linked to enhanced chromatin occupancy and gene activation. Collectively, our findings show that hotspot mutations in a chromatin-reader domain drive self-reinforced recruitment, derailing normal cell-fate control during development and leading to an oncogenic outcome.


Characteristics of hemophilia patients with factor VIII inhibitors detected by prospective screening.
Characteristics of inhibitors identified by prospective screening may differ from those detected clinically. In a prospective study at 17 hemophilia centers with central inhibitor measurement by Nijmegen-Bethesda assay, 23 (2.8%) of 824 hemophilia A patients had new inhibitors detected: nine high-titer inhibitors (HTI: 7 ≥ 5.0 NBU plus 2 of 2.6 and 3.4 NBU at immune tolerance induction initiation) and 14 low-titer inhibitors (LTI: 0.5-1.9 NBU). HTI occurred at an earlier age (median 2 years, range 1-18, vs. median 11 years, range 2-61, P = 0.016). Both HTI (22%) and LTI (43%) occurred in non-severe patients. All HTI, but only 64% of LTI, were found to be FVIII-specific by chromogenic Bethesda assay or fluorescence immunoassay (FLI), indicating a high rate of false-positive LTI. Repeat specimens confirmed all HTI, 7/9 LTI, and 7/7 FVIII-specific LTI. FLI results were similar between HTI and FVIII-specific LTI; all included IgG1 and IgG4 subclasses. A comparable prospective study conducted from 1975 to 1979 at 13 U.S. centers found 31 (2.4%) new inhibitors among 1,306 patients. In both studies, one-third of inhibitors occurred in non-severe patients and one-quarter after 150 exposure days (ED). Significant differences were seen in the age at which inhibitors occurred (median 16 years in the older study vs. 5 years currently, P = 0.024) and in ED before inhibitor development, 10% in the older study and 43% currently study occurring within 20 ED, suggesting a temporal change in inhibitor development. Prospective screening detects inhibitors in patients of all severities, ages, and ED. Some LTI, however, are false positives.


Profiling of victimization, perpetration, and participation: A latent class analysis among people with severe mental illness.
Persons with severe mental illness are more prone to victimization and experience more difficulties regarding societal participation than other community members. These experiences vary greatly among individuals. Community mental health care should offer more individualized support by addressing these differences in experience. Therefore, this study aimed to identify subgroups of outpatients with severe mental illness based on their experiences of social participation and victimization. Data from patients with severe mental illness from eight outpatient teams in the Netherlands were used to perform latent class analysis. From the total caseload, 395 patients agreed to participate. Classes were based on: i) criminal victimization incidents, ii) criminal perpetration incidents (Dutch Safety Monitor), iii) experienced discrimination (DISC-12), and iv) social functioning (Social Functioning Scale). Also, to investigate differences between the classes, socio-demographic, clinical, and person-related variables were examined. Three classes were identified. The Victimized and Perpetrating class (34.4%) had the highest prevalence of discrimination, victimization, and perpetration, and intermediate scores on social functioning subscales. This class also experienced the most problems in other domains, such as psychosocial functioning and quality of life. The Discriminated and Avoiding class (36.4%) had moderate scores for discrimination, victimization and perpetration, and the lowest scores for social functioning and social support. The General Difficulties class (28.8%) had the lowest prevalence of discrimination, victimization, and perpetration, and the highest scores on social functioning. These distinct classes offer new insights to mental health professionals in outpatient teams in in their aim to positively influence the patient's social context during rehabilitation; this includes addressing the role of victimization, and indicates the relevance of distinctive approaches and the support needed for each class. Professionals may need to focus more on the impact of difficulties in their patients' social context to adequately support them in the rehabilitation process.


Promising alternative settings for HPV vaccination of US adolescents.
Vaccination in alternative settings, defined here as being outside of traditional primary care, can help address the pressing public health problem of low human papillomavirus vaccine coverage among adolescents in the United States. Pharmacies are promising because they are highly accessible and have well established immunization practices. However, pharmacies currently face policy and reimbursement challenges. School-located mass vaccination programs are also promising because of their high reach and demonstrated success in providing other vaccines, but control by local policymakers and challenges in establishing community partnerships complicate widespread implementation. Health centers in schools are currently too few to greatly increase access to human papillomavirus vaccine. Specialty clinics have experience with vaccination, but the older age of their patient populations limits their reach. Future steps to making alternative settings a success include expanding their use of statewide vaccine registries and improving their coordination with primary care providers.


Evolutionary history of the Tricladida and the Platyhelminthes: an up-to-date phylogenetic and systematic account.
Within the free-living platyhelminths, the triclads, or planarians, are the best-known group, largely as a result of long-standing and intensive research on regeneration, pattern formation and Hox gene expression. However, the group's evolutionary history has been long debated, with controversies ranging from their phyletic structure and position within the Metazoa to the relationships among species within the Tricladida. Over the the last decade, with the advent of molecular phylogenies, some of these issues have begun to be resolved. Here, we present an up-to-date summary of the main phylogenetic changes and novelties with some comments on their evolutionary implications. The phylum has been split into two groups, and the position of the main group (the Rhabdithophora and the Catenulida), close to the Annelida and the Mollusca within the Lophotrochozoa, is now clear. Their internal relationships, although not totally resolved, have been clarified. Tricladida systematics has also experienced a revolution since the implementation of molecular data. The terrestrial planarians have been demonstrated to have emerged from one of the freshwater families, giving a different view of their evolution and greatly altering their classification. The use of molecular data is also facilitating the identification of Tricladida species by DNA barcoding, allowing better knowledge of their distribution and genetic diversity. Finally, molecular phylogenetic and phylogeographical analyses, taking advantage of recent data, are beginning to give a clear picture of the recent history of the Dugesia and Schmidtea species in the Mediterranean.


Tannins: current knowledge of food sources, intake, bioavailability and biological effects.
Tannins are a unique group of phenolic metabolites with molecular weights between 500 and 30 000 Da, which are widely distributed in almost all plant foods and beverages. Proanthocyanidins and hydrolysable tannins are the two major groups of these bioactive compounds, but complex tannins containing structural elements of both groups and specific tannins in marine brown algae have also been described. Most literature data on food tannins refer only to oligomeric compounds that are extracted with aqueous-organic solvents, but a significant number of non-extractable tannins are usually not mentioned in the literature. The biological effects of tannins usually depend on their grade of polymerisation and solubility. Highly polymerised tannins exhibit low bioaccessibility in the small intestine and low fermentability by colonic microflora. This review summarises a new approach to analysis of extractable and non-extractable tannins, major food sources, and effects of storage and processing on tannin content and bioavailability. Biological properties such as antioxidant, antimicrobial and antiviral effects are also described. In addition, the role of tannins in diabetes mellitus has been discussed.


Involvement of oxidative stress in hydroquinone-induced cytotoxicity in catalase-deficient Escherichia coli mutants.
Hydroquinone is a benzene-derived metabolite. To clarify whether the reactive oxygen species (ROS) are involved in hydroquinone-induced cytotoxicity, we constructed transformants of Escherichia coli (E. coli) strains that express mammalian catalase gene derived from catalase mutant mice (Cs(b), Cs(c)) and the wild-type (Cs(a)) using a catalase-deficient E. coli UM255 as a recipient. Specific catalase activities of these tester strains were in order of Cs(a) > Cs(c) > Cs(b) > UM255, and their susceptibility to hydrogen peroxide (H2O2) showed UM255 > Cs(b) > Cs(c) > Cs(a). We found that hydroquinone exposure reduced the survival of catalase-deficient E. coli mutants in a dose-dependent manner significantly, especially in the strains with lower catalase activities. Hydroquinone toxicity was also confirmed using zone of inhibition test, in which UM255 was the most susceptible, showing the largest zone of growth inhibition, followed by Cs(b), Cs(c) and Cs(a). Furthermore, we found that hydroquinone-induced cell damage was inhibited by the pretreatment of catalase, ascorbic acid, dimethyl sulfoxide (DMSO), and ethylenediaminetetraacetic acid (EDTA), and augmented by superoxide dismutase (both CuZnSOD and MnSOD). The present results suggest that H2O2 is probably involved in hydroquinone-induced cytotoxicity in catalase-deficient E. coli mutants and catalase plays an important role in protection of the cells against hydroquinone toxicity.


The Effects of Different Exercise Trainings on Suprahyoid Muscle Activation, Tongue Pressure Force and Dysphagia Limit in Healthy Subjects.
Suprahyoid muscle activation and tongue pressure force play a critical role for swallowing function. In addition, dysphagia limit is one of most important factors indicating swallowing efficiency. The purpose of this study was to compare the effects of 8-week training sessions of three different exercises including chin tuck against resistance (CTAR), Shaker exercises and chin tuck exercise with theraband on suprahyoid muscle activity, anterior tongue pressure and dysphagia limit in healthy subjects. Thirty-six healthy volunteers aged between 18 and 40 years who scored below 3 points from Turkish version of Eating Assessment Tool (T-EAT-10) were included in the study, and all participants were divided into three groups randomly. Maximal suprahyoid muscle activations and dysphagia limit of participants were assessed by superficial electromyography. CTAR and chin tuck exercise with theraband increased the maximum suprahyoid muscle activation (p1 = 0.004, p2 = 0.018), whereas Shaker exercise did not increase maximal suprahyoid muscle activation (p = 0.507) after exercise training. CTAR and chin tuck exercise with theraband increased tongue pressure (p1 = 0.045, p2 = 0.041), while Shaker exercise did not increase anterior tongue pressure (p = 0.248). There was no statistically significant difference in dysphagia limits in three groups between before and after exercise training (p > 0.05). As a result, although CTAR seems to be the most effective exercise in most parameters, chin tuck exercise with theraband can also be used as an alternative to CTAR to improve suprahyoid muscle activity and tongue pressure.


Immune escape and immunotherapy of acute myeloid leukemia.
In spite of the recent approval of new promising targeted therapies, the clinical outcome of patients with acute myeloid leukemia (AML) remains suboptimal, prompting the search for additional and synergistic therapeutic rationales. It is increasingly evident that the bone marrow immune environment of AML patients is profoundly altered, contributing to the severity of the disease but also providing several windows of opportunity to prompt or rewire a proficient antitumor immune surveillance. In this Review, we present current evidence on immune defects in AML, discuss the challenges with selective targeting of AML cells, and summarize the clinical results and immunologic insights from studies that are testing the latest immunotherapy approaches to specifically target AML cells (antibodies, cellular therapies) or more broadly reactivate antileukemia immunity (vaccines, checkpoint blockade). Given the complex interactions between AML cells and the many components of their environment, it is reasonable to surmise that the future of immunotherapy in AML lies in the rational combination of complementary immunotherapeutic strategies with chemotherapeutics or other oncogenic pathway inhibitors. Identifying reliable biomarkers of response to improve patient selection and avoid toxicities will be critical in this process.


Clinical outcome of a nonvalved Aurolab aqueous drainage implant in posterior segment versus anterior chamber.
To evaluate the outcome of a nonvalved Aurolab aqueous drainage implant (AADI) in the management of refractory glaucoma. Retrospective case series of patients with refractory glaucoma underwent AADI implantation in posterior segment (PS group) or anterior chamber (AC group) with minimum follow-up of 1 year. Primary outcome criterion was success, defined as intraocular pressure (IOP) <18 or >6 mm Hg or IOP reduced to <20% from baseline, for two consecutive visits after 3 months. Failure was defined as inability to meet IOP criteria, any additional glaucoma surgery, loss of light perception, and implant explantation. Secondary outcome criteria compared groups based on mean IOP, mean glaucoma medication use, best-corrected visual acuity, and complications at each postoperative visit. In the AC and PS group of 64 patients, 32 tubes each were placed. Preoperative mean IOP was 37.41 ± 8.6 and 43.38 ± 10.3 mm Hg in AC and PS, respectively. Postoperatively IOP reduced to 14.22 ± 4.9 and 15.21 ± 8.1 mm Hg in AC and PS groups, respectively (P < 0.001). Preoperative mean antiglaucoma medication changed from 2.56 ± 0.9 and 3.44 ± 0.5 to 1.03 ± 0.9 and 1.67 ± 0.5 in AC and PS, respectively, postoperatively (P < 0.001). No significant change in VA was noted in either group. At 12 months, success rate was 84% in AC group and 72% in PS group, with PS group having 2.63 times higher hazard (risk) of failure than AC group. AADI implantation in PS or AC is a safe and effective method for IOP control in refractory glaucoma with its low cost being of significance in developing countries.


Grouping strategies in number estimation extend the subitizing range.
When asked to estimate the number of items in a visual array, educated adults and children are more precise and rapid if the items are clustered into small subgroups rather than randomly distributed. This phenomenon, termed "groupitizing", is thought to rely on the recruitment of the subitizing system (dedicated to the perception of very small numbers), with the aid of simple arithmetical calculations. The aim of current study is to verify whether the advantage for clustered stimuli does rely on subitizing, by manipulating attention, known to strongly affect attention. Participants estimated the numerosity of grouped or ungrouped arrays in condition of full attention or while attention was diverted with a dual-task. Depriving visual attention strongly decreased estimation precision of grouped but not of ungrouped arrays, as well as increasing the tendency for numerosity estimation to regress towards the mean. Additional explorative analyses suggested that calculation skills correlated with the estimation precision of grouped, but not of ungrouped, arrays. The results suggest that groupitizing is an attention-based process that leverages on the subitizing system. They also suggest that measuring numerosity estimation thresholds with grouped stimuli may be a sensitive correlate of math abilities.


Precise long-range synchronization of activity and silence in neocortical neurons during slow-wave oscillations [corrected].
Slow-wave sleep is characterized by alternating periods of activity and silence in corticothalamic networks. Both activity and silence are stable network states, but the mechanisms of their alternation remain unknown. We show, using simultaneous multisite intracellular recordings in cats, that slow rhythm involves all neocortical neurons and that both activity and silence started almost synchronously in cells located up to 12 mm apart. Activity appeared predominantly at the area 5/7 border and spread in both anterior and posterior directions. The activity started earlier in fast-spiking cells and intrinsically bursting cells than in regular-spiking neurons. These results provide direct evidence for two mechanisms of active state generation: spread of activity from a local focus and synchronization of weaker activity, originating at multiple locations. Surprisingly, onsets of silent states were synchronized even more precisely than the onsets of activity, showing no latency bias for location or cell type. This most intriguing finding exposes a major gap in understanding the nature of state alternation. We suggest that it is the synchronous termination of activity and occurrence of silent states of the neuronal network that makes the EEG picture during slow-wave sleep so characteristic. Synchronous onset of silence in distant neurons cannot rely exclusively on properties of individual cells and synapses, such as adaptation of neuronal firing or synaptic depression; instead, it implies the existence of a network mechanism. Revealing this yet unknown large-scale mechanism, which switches network activity to silence, will aid our understanding of the origin of brain rhythms in normal function and pathology.


SU-E-T-147: Film Dosimetry Verification for TSE Using An Epson Scanner.
To commission and verify an Epson scanner for film dosimetry for total skin electron beam therapy (TSEB). Use data from an IBA PPC40 parallel-plate ion chamber and Sun Nuclear QED skin diode detectors as standard; we have made comparisons to the film measurement using Kodak XV films. Hurter-Driffield (HD) curve are established for 6 MeV total skin electron beams at a source-to-surface distance (SSD) of 5 m. Also HD curves are built for 6 MeV at a 100 cm SSD. Dose profiles for a series of oblique incident large electron fields are measured using the film for approximately 80 cGy dose delivered at the peak. The film is then scanned using two scanners, an Epson expression 10000 XL and a Vidar VXR-16 Dosimetry Pro. The optimal scanning conditions (e.g., dot per pixel size, internal color correction scheme) are chosen for the Epson scanner. Matlab is then used to analyze the optical density (OD) of the scanned films. A transmission densitometer made by Tobias Associates transmission is used to analyze the films to give a classical standard. The analysis of the Epson scanner is presented in two forms: one with and one without the HD correction from the established HD curve. The error analysis gives an uncertainty of 5% without the HD correction. An improved result of approximately 3% is found when an HD correction is applied to the analysis. A simple Epson scanner satisfies the commissioning standards for TSEB when an HD curve correction is applied.


RTS,S/AS02A malaria vaccine does not induce parasite CSP T cell epitope selection and reduces multiplicity of infection.
The candidate malaria vaccine RTS,S/AS02A is a recombinant protein containing part of the circumsporozoite protein (CSP) sequence of Plasmodium falciparum, linked to the hepatitis B surface antigen and formulated in the proprietary adjuvant system AS02A. In a recent trial conducted in children younger than age five in southern Mozambique, the vaccine demonstrated significant and sustained efficacy against both infection and clinical disease. In a follow-up study to the main trial, breakthrough infections identified in the trial were examined to determine whether the distribution of csp sequences was affected by the vaccine and to measure the multiplicity of infecting parasite genotypes. P. falciparum DNA from isolates collected during the trial was used for genotype studies. The main trial was carried out in the Manhiça district, Maputo province, Mozambique, between April 2003 and May 2004. Children from the two cohorts of the main trial provided parasite isolates as follows: children from Cohort 1 who were admitted to hospital with clinical malaria; children from Cohort 1 who were parasite-positive in a cross-sectional survey at study month 8.5; children from Cohort 2 identified as parasite-positive during follow-up by active detection of infection. Divergence of DNA sequence encoding the CSP T cell-epitope region sequence from that of the vaccine sequence was measured in 521 isolates. The number of distinct P. falciparum genotypes was also determined. We found no evidence that parasite genotypes from children in the RTS,S/AS02A arm were more divergent than those receiving control vaccines. For Cohort 1 (survey at study month 8.5) and Cohort 2, infections in the vaccine group contained significantly fewer genotypes than those in the control group, (p = 0.035, p = 0.006), respectively, for the two cohorts. This was not the case for children in Cohort 1 who were admitted to hospital (p = 0.478). RTS,S/AS02A did not select for genotypes encoding divergent T cell epitopes in the C-terminal region of CSP in this trial. In both cohorts, there was a modest reduction in the mean number of parasite genotypes harboured by vaccinated children compared with controls, but only among those with asymptomatic infections.


Bioturbation: impact on the marine nitrogen cycle.
Sediments play a key role in the marine nitrogen cycle and can act either as a source or a sink of biologically available (fixed) nitrogen. This cycling is driven by a number of microbial remineralization reactions, many of which occur across the oxic/anoxic interface near the sediment surface. The presence and activity of large burrowing macrofauna (bioturbators) in the sediment can significantly affect these microbial processes by altering the physicochemical properties of the sediment. For example, the building and irrigation of burrows by bioturbators introduces fresh oxygenated water into deeper sediment layers and allows the exchange of solutes between the sediment and water column. Burrows can effectively extend the oxic/anoxic interface into deeper sediment layers, thus providing a unique environment for nitrogen-cycling microbial communities. Recent studies have shown that the abundance and diversity of micro-organisms can be far greater in burrow wall sediment than in the surrounding surface or subsurface sediment; meanwhile, bioturbated sediment supports higher rates of coupled nitrification-denitrification reactions and increased fluxes of ammonium to the water column. In the present paper we discuss the potential for bioturbation to significantly affect marine nitrogen cycling, as well as the molecular techniques used to study microbial nitrogen cycling communities and directions for future study.


Study of in vitro RBCs membrane elasticity with AOD scanning optical tweezers.
The elasticity of red cell membrane is a critical physiological index for the activity of RBC. Study of the inherent mechanism for RBCs membrane elasticity transformation is attention-getting all along. This paper proposes an optimized measurement method of erythrocytes membrane shear modulus incorporating acousto-optic deflector (AOD) scanning optical tweezers system. By use of this method, both membrane shear moduli and sizes of RBCs with different in vitro times were determined. The experimental results reveal that the RBCs membrane elasticity and size decline with in vitro time extension. In addition, semi quantitative measurements of S-nitrosothiol content in blood using fluorescent spectrometry during in vitro storage show that RBCs membrane elasticity change is positively associated with the S-nitrosylation level of blood. The analysis considered that the diminished activity of the nitric oxide synthase makes the S-nitrosylation of in vitro blood weaker gradually. The main reason for worse elasticity of the in vitro RBCs is that S-nitrosylation effect of spectrin fades. These results will provide a guideline for further study of in vitro cells activity and other clinical applications.


Moving-Tolerant Augmented Reality Surgical Navigation System Using Autostereoscopic Three-Dimensional Image Overlay.
Augmented reality (AR) surgical navigation systems based on image overlay have been used in minimally invasive surgery. However, conventional systems still suffer from a limited viewing zone, a shortage of intuitive three-dimensional (3D) image guidance and cannot be moved freely. To fuse the 3-D overlay image with the patient in situ, it is essential to track the overlay device while it is moving. A direct line-of-sight should be maintained between the optical markers and the tracker camera. In this study, we propose a moving-tolerant AR surgical navigation system using autostereoscopic image overlay, which can avoid the use of the optical tracking system during the intraoperative period. The system captures binocular image sequences of environmental change in the operation room to locate the overlay device, rather than tracking the device directly. Therefore, it is no longer required to maintain a direct line-of-sight between the tracker and the tracked devices. The movable range of the system is also not limited by the scope of the tracker camera. Computer simulation experiments demonstrate the reliability of the proposed moving-tolerant AR surgical navigation system. We also fabricate a computer-generated integral photography-based 3-D overlay AR system to validate the feasibility of the proposed moving-tolerant approach. Qualitative and quantitative experiments demonstrate that the proposed system can always fuse the 3-D image with the patient, thus, increasing the feasibility and reliability of traditional 3-D overlay image AR surgical navigation systems.


Management of Huntington's disease: role of tetrabenazine.
Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by progressive involuntary movements, neuropsychiatric disturbances, and cognitive impairment. The use of tetrabenazine (TBZ), a specific inhibitor of vesicular monoamine transporter, is approved for chorea in HD patients. We aimed to review the medical literature concerning the efficacy and tolerability of TBZ in the treatment of HD patients and to report our personal experience about TBZ use in a cohort of HD patients. We searched PubMed (1960 to July 2010) using the following keywords: "tetrabenazine" + "huntington's disease + chorea". We included randomized controlled trials, open-label trials, and retrospective studies. We excluded case reports and studies conducted on fewer than 20 patients. In addition, we retrospectively evaluated 2 years' follow-up of TBZ treatment on motor and cognitive performances and functional abilities in 28 HD patients, compared with 10 patients treated by other neuroleptics (clotiapine). Only four papers fulfilled the requested criteria. In the first study, which included 84 randomized outpatients, TBZ showed a significant improvement of chorea compared with placebo. In the open-label study extension, TBZ confirmed its efficacy on chorea, with a frequent occurrence of withdrawals due to side effects. In a retrospective study of long-term efficacy, 63 patients under TBZ therapy for an average period of 34 months showed a stable effect on chorea, despite a slight reduction of effect over time. In a telephone survey conducted on a total of 118 patients affected by different movement disorders, TBZ showed the most favorable effect for the 28 included HD patients. Our HD patients showed a slight deterioration of motor performances over time that was nonsignificant compared with TBZ or clotiapine treatments. Despite the fact that the global effect of TBZ seems positive in HD, more attention on evaluating symptomatic treatments for cognitive and psychiatric deterioration as well as motor deterioration would alleviate this devastating disorder until a neuroprotective treatment becomes available.


Effect of morphine on thioglycollate-induced peritonitis in chickens.
Morphine exerts immunomodulatory effects dependent on several factors including species and parameter examined. The aim of this study was to assess the influence of morphine on experimental peritonitis and leukocyte activity in young chickens of both sexes. Peritonitis was elicited by intraperitoneal injection of thioglycollate (TG) alone or supplemented with morphine; additional chicken groups were injected with morphine alone. Morphine-treated birds (with or without elicited peritonitis) were pretreated with naltrexone, an antagonist of opioid receptors. Control groups were intact or PBS-injected. At specific postinjection intervals peritoneal leukocytes (PTL) were obtained by flushing the peritoneal cavity, counted and used for in vitro assays of activity (respiratory burst). Vascular permeability was measured using Evans blue solution. Inflammatory reaction and morphine influence were gender-dependent: the extent of TG-induced peritonitis was higher in males and it was additionally stimulated by morphine. This pro-inflammatory effect of morphine was not seen in females. PTL collected from morphine-treated chickens of both sexes exhibited a stronger nitroblue tetrazolium reduction than in non-treated birds and this effect was antagonized by naltrexone. The effect of morphine on peritonitis in chickens appears different to that in other vertebrate species, although the mechanism(s) of its influence on leukocyte activity are similar.


Electrochemically monitoring the antibiotic susceptibility of Pseudomonas aeruginosa biofilms.
The condition of cells in Pseudomonas aeruginosa biofilms was monitored via the electrochemical detection of the electro-active virulence factor pyocyanin in a fabricated microfluidic growth chamber coupled with a disposable three electrode cell. Cells were exposed to 4, 16, and 100 mg L(-1) colistin sulfate after overnight growth. At the end of testing, the measured maximum peak current (and therefore pyocyanin concentration) was reduced by approximately 68% and 82% in P. aeruginosa exposed to 16 and 100 mg L(-1) colistin sulfate, respectively. Samples were removed from the microfluidic chamber, analyzed for viability using staining, and streaked onto culture plates to confirm that the P. aeruginosa cells were affected by the antibiotics. The correlation between electrical signal drop and the viability of P. aeruginosa cells after antibiotic exposure highlights the usefulness of this approach for future low cost antibiotic screening applications.


Mitochondrial DNA sequences in ancient Australians: Implications for modern human origins.
DNA from ancient human remains provides perspectives on the origin of our species and the relationship between molecular and morphological variation. We report analysis of mtDNA from the remains of 10 ancient Australians. These include the morphologically gracile Lake Mungo 3 [ approximately 60 thousand years (ka) before present] and three other gracile individuals from Holocene deposits at Willandra Lakes (<10 ka), all within the skeletal range of living Australians, and six Pleistocene/early Holocene individuals (15 to <8 ka) from Kow Swamp with robust morphologies outside the skeletal range of contemporary indigenous Australians. Lake Mungo 3 is the oldest (Pleistocene) "anatomically modern" human from whom DNA has been recovered. His mtDNA belonged to a lineage that only survives as a segment inserted into chromosome 11 of the nuclear genome, which is now widespread among human populations. This lineage probably diverged before the most recent common ancestor of contemporary human mitochondrial genomes. This timing of divergence implies that the deepest known mtDNA lineage from an anatomically modern human occurred in Australia; analysis restricted to living humans places the deepest branches in East Africa. The other ancient Australian individuals we examined have mtDNA sequences descended from the most recent common ancestor of living humans. Our results indicate that anatomically modern humans were present in Australia before the complete fixation of the mtDNA lineage now found in all living people. Sequences from additional ancient humans may further challenge current concepts of modern human origins.


Adjudicating outcomes: fundamentals.
The adjudication of outcomes has rarely been reported in the orthopaedic literature, although this process is commonly used and reported in clinical trials of other medical disciplines. Adjudication of outcomes provides more reliable and valid outcome assessment, especially when the outcome is subjective as in the case of fracture-healing. The successful implementation of adjudication in a clinical trial is an important and complex process. The process requires a substantial infrastructure of research personnel to oversee data collection at the clinical sites. The development of an adjudication charter specific to the study is a critical aspect of adjudication as it outlines the adjudication committee membership as well as their roles and responsibilities and defines the adjudication process and the decision rules. Web-based adjudication has facilitated the process as it allows rapid, efficient, and timely adjudication. This article provides an overview of the adjudication process, along with details on the common pearls and pitfalls associated with this method of outcomes assessment.


Robotic central pancreatectomy with pancreaticogastrostomy (transgastric approach) in a solid pseudopapillary tumor of the pancreas.
Function preserving minimal invasive pancreatectomy is thought to be ideal approach for benign and borderline malignant tumors of the pancreas because these tumors can be expected for longterm survival. Pancreatic tumor in the neck of the pancreas is a challenging issue. Pancreaticoduodenectomy and distal pancreatectomy with/without splenectomy are the usual modes of surgery for them. Central pancreatectomy is a rare surgical procedure, even in open surgery. With the development of laparoscopic experiences, a few cases of laparoscopic central pancreatectomy have begun to be reported, but they are believed to be still demanding tremendous laparoscopic skills and experiences. However, advancements in technology have encouraged surgeons to overcome the limitations of conventional laparoscopic surgery. Herein, we report a case of robot-assisted central pancreatectomy with pancreaticogastrostomy (transgastric approach). Wrist-like movements of effector instruments and stable 3-D visualization provided by the robot surgical system are believed to enhance the precise and safe laparoscopic performance. More experiences need to be accumulated in order to evaluate the real value of robot pancreatic surgery.


Lateral flow assays for Ochratoxin A using metal nanoparticles: comparison of "adsorption-desorption" approach to linkage inversion assembled nano-aptasensors (LIANA).
Nano-aptamer probes were prepared and used in lateral flow colorimetric assays for the detection of Ochratoxin A (OTA). In this study, two approaches were examined using 5'-biotin-modified OTA aptamers and silver or gold nanoparticles (AgNP or AuNP). The first method used an "adsorption-desorption" approach wherein aptamers were adsorbed onto the metal nanoparticle surface. Upon the addition of OTA, the aptamer binds specifically to the target, releasing the NPs. The above solutions were applied on a lateral flow assay (LFA) and a detection limit of 6.3 nM was achieved with both metal nanoparticles. The second method used a labelled approach based on Linkage Inversion Assembled Nano-Aptasensors (LIANAs) using a DNA linker containing a 5'-5' linkage inversion (5'-5' linker) to assemble biotinylated aptamer-functionalized metal nanoparticles. In the presence of target, OTA specifically binds with its aptamer leading to release of the linker and disassembly of LIANA aggregates into dispersed nanoparticles. The same solutions were applied in LFA format and the lowest detection limit of 0.63 nM was achieved. The results indicated that the LIANA-based LFA strips were more sensitive than the "adsoprtion-desorption" LFAs. Both lateral flow assays are inexpensive, simple, and rapid to perform and produces results visible to the naked-eye.


Randomized phase III trial of ABVD versus Stanford V with or without radiation therapy in locally extensive and advanced-stage Hodgkin lymphoma: an intergroup study coordinated by the Eastern Cooperative Oncology Group (E2496).
Although ABVD (doxorubicin, bleomycin, vinblastine, and dacarbazine) has been established as the standard of care in patients with advanced Hodgkin lymphoma, newer regimens have been investigated, which have appeared superior in early phase II studies. Our aim was to determine if failure-free survival was superior in patients treated with the Stanford V regimen compared with ABVD. The Eastern Cooperative Oncology Group, along with the Cancer and Leukemia Group B, the Southwest Oncology Group, and the Canadian NCIC Clinical Trials Group, conducted this randomized phase III trial in patients with advanced Hodgkin lymphoma. Stratification factors included extent of disease (localized v extensive) and International Prognostic Factors Project Score (0 to 2 v 3 to 7). The primary end point was failure-free survival (FFS), defined as the time from random assignment to progression, relapse, or death, whichever occurred first. Overall survival, a secondary end point, was measured from random assignment to death as a result of any cause. This design provided 87% power to detect a 33% reduction in FFS hazard rate, or a difference in 5-year FFS of 64% versus 74% at two-sided .05 significance level. There was no significant difference in the overall response rate between the two arms, with complete remission and clinical complete remission rates of 73% for ABVD and 69% for Stanford V. At a median follow-up of 6.4 years, there was no difference in FFS: 74% for ABVD and 71% for Stanford V at 5 years (P = .32). ABVD remains the standard of care for patients with advanced Hodgkin lymphoma.


Testing a Novel Device for Accurate Ultrasound Delivery During Crystalline Lens Phacoemulsification Surgery.
To assess whether the use of a patented, novel feedback device intended to accurately control phacoemulsification tip elongation is effective under varying machine settings and material resistance. Sculpt mode phaco (550-mm Hg Venturi pump; elongations, 35 and 70 µm) and quadrant settings (550-mm Hg Venturi pump; elongations, 15, 30, and 60 µm) were used in agar gel of incremental density (1%, 2%, 3%, and 6% in demineralized water). Dispersed lens fragments were also simulated with 6% agar gel spherules (2-5 mm in diameter; 550-mm Hg vacuum, and 60-µm elongation). Actual phaco tip elongation was measured on voltage readings from the piezoelectric crystals and compared to nominal elongation with feedback control off and on. Mismatch between nominal and actual elongation when feedback control was off in sculpt mode varied between -13.51 µm and -23.07 µm of nominal elongation; in quadrant mode, mismatch varied between -2.79 µm and -20.41 µm. When the feedback control system was switched on, mismatch varied between -0.02 µm and +0.43 µm (P < 0.001 for all matchings). When the feedback system was off, the elongation mismatch among the 1%, 3%, and 6% agar was also statistically significant (P < 0.001). Elongation was 44.72 ± 4.16 µm with feedback control off and 60.02 ± 1.63 µm with it on (nominal elongation 60 µm; P < 0.001) when emulsifying agar 6% gel fragments. Dispersion of elongation data was also significantly wider when feedback control was turned off. A novel feedback control system effectively controls elongation accuracy regardless of the resistance offered by incremental agar gel concentrations. Implementing feedback control in phaco handpieces dramatically improves surgical accuracy. The translational value of this research relies on its immediate applicability to routine cataract surgery, resulting in a more appropriate use of ultrasound energy.


LYSP100-associated nuclear domains (LANDs): description of a new class of subnuclear structures and their relationship to PML nuclear bodies.
The PML gene is fused to the retinoic acid receptor alpha (RAR alpha) gene in t(15;17) acute promyelocytic leukemia (APL), creating a PML-RAR alpha fusion oncoprotein. The PML gene product has been localized to subnuclear dot-like structures variously termed PODs, ND10s, Kr bodies, or PML nuclear bodies (PML NBs). The present study describes the cloning of a lymphoid-restricted gene, LYSP100, that is homologous to another protein that localizes to PML NBs, SP100. In addition to SP100 homology regions, one LYSP100 cDNA isoform contains a bromodomain and a PHD/TTC domain, which are present in a variety of transcriptional regulatory proteins. By immunofluorescence, LYSP100 was localized to nuclear dots that were surprisingly largely nonoverlapping with PML NBs. However, a minority of LYSP100 nuclear dots exactly colocalized with PML and SP100. We term the LYSP100 structures "LANDs," for LYSP100-associated nuclear domains. Although LYSP100 is expressed only in lymphoid cells, LANDs could be visualized in HeLa cells by transfection of a LYSP100 cDNA. Immunoelectron microscopy revealed LANDs to be globular, electron-dense structures morphologically distinct from the annular structures characteristic of PML NBs. LANDs were most often found in the nucleoplasm, but were also found at the nuclear membrane and in the cytoplasm, suggesting that these structures may traffic between the cytoplasm and the nucleus. By double-immunogold labeling of PML and LYSP100, some LANDs were shown to contain both PML and LYSP100. Thus, PML is localized to a second subnuclear domain that is morphologically and biochemically distinct from PML NBs.


Spontaneous synchronous synaptic calcium transients in cultured cortical neurons.
The firing pattern displayed by neuronal aggregates is thought to play a key role in cortical development and physiology. In this study, we have employed optical recording of intracellular calcium to monitor activity of multiple neurons simultaneously in primary cortical cultures. With this approach, we have observed spontaneous synchronous calcium transients among adjacent cortical neurons. These transients appear to be mediated by prominent spontaneous synaptic excitation, as they are enhanced by picrotoxin, a blocker of inhibitory GABAergic transmission, and reduced by antagonism of glutamate receptors or addition of TTX. After picrotoxin treatment, the calcium transients exhibit regular frequency and amplitude, and occur in synchrony with bursts of excitatory synaptic potentials every 10-20 sec. Using electrical stimulation, we have identified a relative refractory period, extending up to 5 sec after a synchronous burst, that may play a role in cell synchronization. NMDA receptor antagonists or reduced extracellular calcium levels lower the amplitude of the calcium transients yet fail to alter their frequency, suggesting that intracellular calcium levels may not be a major determinant of burst frequency. In contrast, mild depolarization with kainic acid (0.5-1 microM) increased burst frequency up to fivefold, suggesting a critical dependence of rhythmic activity on membrane potential. Chronic blockade of electrical activity with TTX beginning a few days after plating of cultures dampens the amplitude and significantly increases the frequency of calcium transients in mature cultures. These studies demonstrate that aggregates of cultured cortical neurons express synchronous firing activity in vitro and that this network activity is dependent in part on neuronal firing during development.


Magnetic stimulation of the left visual cortex impairs expert word recognition.
One of the hallmarks of expert reading is the ability to identify arrays of several letters quickly and in parallel. Such length-independent reading has only been found for word stimuli appearing in the right visual hemifield (RVF). With left hemifield presentation (LVF), response times increase as a function of word length. Here we investigated the comparative efficiency with which the two hemispheres are able to recognize visually presented words, as measured by word length effects. Repetitive transcranial magnetic stimulation (rTMS) of the left occipital cortex disrupted expert processing of the RVF such that a length effect was created (Experiment 1). Right occipital rTMS, on the other hand, had no such effect on RVF words and nor did it modulate the length effect already present in the LVF. Experiment 2 explored the time course of these TMS-induced effects by applying single pulses of TMS at various stimulus-onset asynchronies for the same task. We replicated the TMS-induced length effect for RVF words, but only when a single pulse was applied to the left visual cortex 80 msec after target presentation. This is the first demonstration of TMS-induced impairment producing a word length effect, and as such confirms the specialization of the left hemisphere in word recognition. It is likely that anatomical differences in the pathway linking retinal input to higher level cortical processing drive this effect.


Small chemical chromatin effectors alter secondary metabolite production in Aspergillus clavatus.
The filamentous fungus Aspergillus clavatus is known to produce a variety of secondary metabolites (SM) such as patulin, pseurotin A, and cytochalasin E. In fungi, the production of most SM is strongly influenced by environmental factors and nutrients. Furthermore, it has been shown that the regulation of SM gene clusters is largely based on modulation of a chromatin structure. Communication between fungi and bacteria also triggers chromatin-based induction of silent SM gene clusters. Consequently, chemical chromatin effectors known to inhibit histone deacetylases (HDACs) and DNA-methyltransferases (DNMTs) influence the SM profile of several fungi. In this study, we tested the effect of five different chemicals, which are known to affect chromatin structure, on SM production in A. clavatus using two growth media with a different organic nitrogen source. We found that production of patulin was completely inhibited and cytochalasin E levels strongly reduced, whereas growing A. clavatus in media containing soya-derived peptone led to substantially higher pseurotin A levels. The HDAC inhibitors valproic acid, trichostatin A and butyrate, as well as the DNMT inhibitor 5-azacytidine (AZA) and N-acetyl-D-glucosamine, which was used as a proxy for bacterial fungal co-cultivation, had profound influence on SM accumulation and transcription of the corresponding biosynthetic genes. However, the repressing effect of the soya-based nitrogen source on patulin production could not be bypassed by any of the small chemical chromatin effectors. Interestingly, AZA influenced some SM cluster genes and SM production although no Aspergillus species has yet been shown to carry detectable DNA methylation.


Alpha+ -thalassaemia and pregnancy in a malaria endemic region of Papua New Guinea.
The effect of maternal alpha+ -thalassaemia on pregnancy was assessed in the north coastal region of Papua New Guinea (PNG), where malaria is hyperendemic and alpha+ -thalassaemia is extremely common. In a prospective study of 987 singleton hospital deliveries, we correlated maternal alpha-globin genotype with markers of reproductive fitness (age in primigravidae, gravidity, pregnancy interval and the number of miscarriages and stillbirths), Plasmodium falciparum(P. falciparum) infection of the mother and placenta, maternal haemoglobin, preterm delivery and birthweight. The frequency of the -alpha genotype in mothers was 0.61. Markers of reproductive fitness were similar in women with and without alpha+ -thalassaemia. Median haemoglobin concentration during pregnancy and after delivery was about 1.0 g/dl lower in homozygous alpha+ -thalassaemia than in women with a normal alpha- globin genotype (P < or = 0.001). The frequency of placental P. falciparum infection and systemic malaria infection after delivery showed no consistent relationship to alpha-globin genotype. The frequency of preterm delivery and low birthweight did not vary significantly according to maternal alpha-globin genotype. Maternal alpha+ -thalassaemia does not affect reproductive fitness or susceptibility to malaria during pregnancy. Although median haemoglobin concentration was significantly lower in mothers homozygous for alpha+ -thalassaemia than those with a normal alpha-globin genotype, this did not result in an adverse outcome of pregnancy.


Direct and indirect effects of exposure to 900 MHz GSM radiofrequency electromagnetic fields on CHO cell line: Evidence of bystander effect by non-ionizing radiation.
The rapid rise in global concerns about the adverse health effects of exposure to radiofrequency radiation (RFR) generated by common devices such as mobile phones has prompted scientists to further investigate the biological effects of these environmental exposures. Non-targeted effects (NTEs) are responses which do not need a direct exposure to be expressed and are particularly significant at low energy radiations. Although NTEs of ionizing radiation are well documented, there are scarcely any studies on non-targeted responses such as bystander effect (BE) after exposure to non-ionizing radiation. The main goal of this research is to study possible RFR-induced BE. Chinese hamster ovary cells were exposed to 900 MHz GSM RFR at an average specific absorption rate (SAR) of 2 W/kg for 4, 12 and 24 hours (h). To generate a uniformly distributed electromagnetic field and avoid extraneous RF exposures a cavity was desined and used. Cell membrane permeability, cell redox activity, metabolic and mitotic cell death and DNA damages were analyzed. Then the most effective exposure durations and statistically significant altered parameters were chosen to assess the induction of BE through medium transfer procedure. Furthermore, intra and extra cellular reactive oxygen species (ROS) levels were measured to assess the molecular mechanism of BE induced by non-ionizing radiation. No statistically significant alteration was found in cell membrane permeability, cell redox activity, metabolic cell activity and micronuclei (MN) frequency in the cells directly exposed to RFR for 4, 12, or 24 h. However, RFR exposure for 24 h caused a statistically significant decrease in clonogenic ability as well as a statistically significant increase in olive moment in both directly exposed and bystander cells which received media from RFR-exposed cells (conditioned culture medium; CCM). Exposure to RFR also statistically significant elevated both intra and extra cellular levels of ROS. Our observation clearly indicated the induction of BE in cells treated with CCM. To our knowledge, this is the first report that a non-ionizing radiation (900 MHz GSM RFR) can induce bystander effect. As reported for ionizing radiation, our results proposed that ROS can be a potential molecule in indirect effect of RFR. On the other hand, we found the importance of ROS in direct effect of RFR but in different ways.


Cromakalim dilates rat cerebral arteries in vitro.
Using an in vitro perfusion method, we examined the effects of cromakalim, a potassium channel opener, on the superior cerebellar arteries of 24 rats. Cromakalim had no effect on contractions induced by 129 mM K+ until a concentration of 10(-5) M was reached. Contractions evoked by 10(-5) M serotonin were attenuated by cromakalim in a concentration-dependent manner (p less than 0.05). The diameter of untreated superior cerebellar arteries remained almost constant with increasing perfusion pressure. However, in the presence of cromakalim vessel diameter increased with increasing perfusion pressure. At concentrations of 3 x 10(-6) M, cromakalim also inhibited basal myogenic tone and dilated unstimulated arteries (p less than 0.01). These results suggest that cromakalim is a cerebrovascular dilator acting on both receptor-mediated and myogenic contractions.


Coordination, framing and innovation: the political sophistication of public health advocates in Ireland.
This study explores the role of the public health advocacy coalition in alcohol policy development in Ireland. Compared with industry actors, much less is known about the membership, activities and influence of public health advocates in alcohol policymaking. To address this gap, this paper identifies several advocacy strategies, drawn from the advocacy coalition framework and other policy theories, and then analyses them in the context of recent Irish developments. The study used theory-building process-tracing to construct a record of the public health advocacy coalition and its campaign to promote the Public Health (Alcohol) Act 2018 in Ireland. Specifically, we drew upon 131 primary documents produced by advocates, 464 newspaper articles and 18 semi-structured interviews with key advocates, public health experts and elected officials to undertake a thematic analysis. Public health advocates in Ireland have developed sophisticated political strategies to foster major alcohol policy change. First, public health advocates led the formation of a broad-based advocacy coalition that helped members to effectively pool their limited resources as well as coordinate their strategy and messaging. Secondly, issue-framing and message discipline played a key role in the coalition's success. Advocates strategically focused upon the policy problem, specifically health harms, rather than the detailed content of the proposed measures. Finally, there is evidence of political learning, where advocates' prior experiences and knowledge of the political system in Ireland spurred innovations in campaigning. These strategies were interdependent and mutually reinforcing, and succeeded in building support for public health advocates' preferred policies among politicians and the general public. There are distinct capabilities that public health actors can mobilize in the policy process to win alcohol policy debates and capitalize on the constraints on industry influence on alcohol policymaking.


Heme as a magnificent molecule with multiple missions: heme determines its own fate and governs cellular homeostasis.
Heme is a prosthetic group of various types of proteins, such as hemoglobin, myoglobin, cytochrome c, cytochrome p450, catalase and peroxidase. In addition, heme is involved in a variety of biological events by modulating the function or the state of hemoproteins. For example, protein synthesis is inhibited in erythroid cells under heme deficiency, as the consequence of the activation of heme-regulated inhibitor (HRI). Iron concentration in the cell is sensed and regulated by the heme-mediated oxidization and subsequent degradation of iron regulatory protein 2 (IRP2). Heme also binds to certain types of potassium channels, thereby inhibiting transmembrane K(+) currents. Importantly, heme determines its own fate; namely, heme regulates its synthesis and degradation through the feedback mechanisms, by which intracellular heme level is precisely maintained. Heme reduces heme synthesis by suppressing the expression of non-specific 5-aminolevulinate synthase (ALAS1) and stimulates heme breakdown by inducing heme oxygenase (HO)-1 expression. ALAS1 and HO-1 are the rate limiting enzymes in heme biosynthesis and catabolism, respectively. Accordingly, under the heme-rich condition, heme binds to cysteine-proline (CP) motifs of ALAS1 and those of transcriptional repressor Bach1, thereby leading to repression of mitochondrial transport of ALAS1 and induction of HO-1 transcription, respectively. Moreover, chemosensing functions of HO-2 containing CP motifs, another isozyme of HO, have been unveiled recently. In this review article, we summarize and update the pleiotropic effects of heme on various biological events and the regulatory network of heme biosynthesis and catabolism.


T lymphocytes specific for immunoglobulin allotype. II. Cloned Igh-1b-specific cytotoxic T cells.
This study describes long-term-cultured lines and clones of cytotoxic T cells (Tc) with specificity for determinants of the Igh-1(b) immunoglobulin allotype. These Tc clones were initiated by repeated stimulation of immune spleen cells from BALB/c mice with an Igh-1(b)-producing myeloma, and then they were maintained in medium supplemented with mitogen-induced growth factors in the absence offurther antigenic stimulation . The lytic potency of these clones was 30-100-fold greater than the primary cultures from which they were derived, and specificity studies showed them to be lytic for Igh-1(b) targets and not for targets expressing Igh-1(a) or Igh-4(b), nor the lipopolysaccharide blasts . Finally, soluble preparations of Ig were tested for their ability to block lysis of labeled Igh-1(b)-expressing targets. The results showed that Igh-1(b) and not other immunoglobulin allotypes or isotypes could block lysis, and that the mechanism of lytic inhibition is due to Igh-1(b)-induced autolysis of the killer cells.


cytoHubba: identifying hub objects and sub-networks from complex interactome.
Network is a useful way for presenting many types of biological data including protein-protein interactions, gene regulations, cellular pathways, and signal transductions. We can measure nodes by their network features to infer their importance in the network, and it can help us identify central elements of biological networks. We introduce a novel Cytoscape plugin cytoHubba for ranking nodes in a network by their network features. CytoHubba provides 11 topological analysis methods including Degree, Edge Percolated Component, Maximum Neighborhood Component, Density of Maximum Neighborhood Component, Maximal Clique Centrality and six centralities (Bottleneck, EcCentricity, Closeness, Radiality, Betweenness, and Stress) based on shortest paths. Among the eleven methods, the new proposed method, MCC, has a better performance on the precision of predicting essential proteins from the yeast PPI network. CytoHubba provide a user-friendly interface to explore important nodes in biological networks. It computes all eleven methods in one stop shopping way. Besides, researchers are able to combine cytoHubba with and other plugins into a novel analysis scheme. The network and sub-networks caught by this topological analysis strategy will lead to new insights on essential regulatory networks and protein drug targets for experimental biologists. According to cytoscape plugin download statistics, the accumulated number of cytoHubba is around 6,700 times since 2010.