Shear bond strength, microleakage, and confocal studies of 4 amalgam alloy bonding agents.
This study was undertaken to determine the relative shear bond strengths and microleakage of 4 bonding agents to dentin and amalgam and to investigate the bonding to dentin through confocal laser scanning microscopy. Sixty noncarious molars were restored with 1 of 4 different systems, and the shear bond strengths were determined. For the microleakage study, Class V amalgam restorations were placed in 60 noncarious teeth. The specimens were thermocycled, the teeth were sectioned, and dye penetration was assessed. For confocal examinations, the first component of the bonding adhesives was labeled with rhodamine B. Each of the adhesives was applied to 3 dentin specimens, which were examined under a confocal laser scanning microscope Of the 4 restorative systems tested, AmalgamBond Plus + HPA and Prime & Bond 2.1/base-catalyst showed significantly higher shear bond strengths. Prime & Bond 2.1/base-catalyst had the lowest microleakage value, which was significantly lower than that of AmalgamBond Plus + HPA and AmalgamBond Plus. AmalgamBond Plus + HPA had the highest variation in both the bond strength and microleakage values. Confocal laser scanning microscopy revealed tag formation, penetration of the bonding agents deep into the tubules, and hybrid layer formation for all 4 bonding systems. The 2 best systems, Prime & Bond 2.1/base-catalyst and, to a lesser extent, AmalgamBond Plus + HPA, utilized a supplemental bonding agent.


Temporal Variation of the Presence of Rhodnius prolixus (Hemiptera: Reduviidae) Into Rural Dwellings in the Department of Casanare, Eastern Colombia.
Rhodnius prolixus (Stål, 1859) is the major vector of Trypanosoma cruzi in Colombia and Venezuela. The species is strongly associated with high-altitude ecotopes, such as sylvatic palms (Attalea butyracea), where spatially and temporally stable infestations are established. We investigated temporal variation in regards to the presence of R. prolixus in rural dwellings in the department of Casanare (eastern Colombia) over a period of 12 mo. Thirty houses were sampled from January to December 2017 by installing Maria sensors, collecting triatomines through community entomological surveillance, and conducting a monthly search in each house. The collection of specimens from the houses varied significantly by month with the higher number of collections occurring in the low-rainfall season and the lower number of collections occurring in the months of increased precipitation. The proportions of males, females, and nymphs also varied significantly throughout the time period: nymphs (fifth instar only) were reported only during May, July, and September and significantly greater numbers of females than males were reported in the inspected dwellings in all months. Density, crowding, and colonization indices varied according to the season. A bloodmeal analysis revealed 17 different hosts. A total of 42 randomly selected R. prolixus specimens were subjected to molecular analyses for detection of T. cruzi DNA with 22 found positive (infection prevalence of 52%). In conclusion, we observed a high presence of R. prolixus (infected with T. cruzi) in dwellings close to native palm plantations. These findings indicate a high risk of vector transmission of T. cruzi for people in the study areas and challenges for the current vector control schemes in the region.


Molecular docking screening using agonist-bound GPCR structures: probing the A2A adenosine receptor.
Crystal structures of G protein-coupled receptors (GPCRs) have recently revealed the molecular basis of ligand binding and activation, which has provided exciting opportunities for structure-based drug design. The A2A adenosine receptor (A2AAR) is a promising therapeutic target for cardiovascular diseases, but progress in this area is limited by the lack of novel agonist scaffolds. We carried out docking screens of 6.7 million commercially available molecules against active-like conformations of the A2AAR to investigate whether these structures could guide the discovery of agonists. Nine out of the 20 predicted agonists were confirmed to be A2AAR ligands, but none of these activated the ARs. The difficulties in discovering AR agonists using structure-based methods originated from limited atomic-level understanding of the activation mechanism and a chemical bias toward antagonists in the screened library. In particular, the composition of the screened library was found to strongly reduce the likelihood of identifying AR agonists, which reflected the high ligand complexity required for receptor activation. Extension of this analysis to other pharmaceutically relevant GPCRs suggested that library screening may not be suitable for targets requiring a complex receptor-ligand interaction network. Our results provide specific directions for the future development of novel A2AAR agonists and general strategies for structure-based drug discovery.


S6K1 and mTOR regulate Rac1-driven platelet activation and aggregation.
Platelet activation and thrombus formation are under the control of signaling systems that integrate cellular homeostasis with cytoskeletal dynamics. Here, we identify a role for the ribosome protein S6 kinase (S6K1) and its upstream regulator mTOR in the control of platelet activation and aggregate formation under shear flow. Platelet engagement of fibrinogen initiated a signaling cascade that triggered the activation of S6K1 and Rac1. Fibrinogen-induced S6K1 activation was abolished by inhibitors of Src kinases, but not Rac1 inhibitors, demonstrating that S6K1 acts upstream of Rac1. S6K1 and Rac1 interacted in a protein complex with the Rac1 GEF TIAM1 and colocalized with actin at the platelet lamellipodial edge, suggesting that S6K1 and Rac1 work together to drive platelet spreading. Pharmacologic inhibitors of mTOR and S6K1 blocked Rac1 activation and prevented platelet spreading on fibrinogen, but had no effect on Src or FAK kinase activation. mTOR inhibitors dramatically reduced collagen-induced platelet aggregation and promoted the destabilization of platelet aggregates formed under shear flow conditions. Together, these results reveal novel roles for S6K1 and mTOR in the regulation of Rac1 activity and provide insights into the relationship between the pharmacology of the mTOR system and the molecular mechanisms of platelet activation.


Cytosolic Access of Intracellular Bacterial Pathogens: The Shigella Paradigm.
Shigella is a Gram-negative bacterial pathogen, which causes bacillary dysentery in humans. A crucial step of Shigella infection is its invasion of epithelial cells. Using a type III secretion system, Shigella injects several bacterial effectors ultimately leading to bacterial internalization within a vacuole. Then, Shigella escapes rapidly from the vacuole, it replicates within the cytosol and spreads from cell-to-cell. The molecular mechanism of vacuolar rupture used by Shigella has been studied in some detail during the recent years and new paradigms are emerging about the underlying molecular events. For decades, bacterial effector proteins were portrayed as main actors inducing vacuolar rupture. This includes the effector/translocators IpaB and IpaC. More recently, this has been challenged and an implication of the host cell in the process of vacuolar rupture has been put forward. This includes the bacterial subversion of host trafficking regulators, such as the Rab GTPase Rab11. The involvement of the host in determining bacterial vacuolar integrity has also been found for other bacterial pathogens, particularly for Salmonella. Here, we will discuss our current view of host factor and pathogen effector implications during Shigella vacuolar rupture and the steps leading to it.


Malpractice: an ogre on the horizon.
Although social workers are sued for malpractice only infrequently, the possibility exists nonetheless. Drawing analogies with areas of liability that exist in other professions, particularly psychiatry, the author calls attention to ways in which the social worker can protect himself against malpractice suits.


Strong purifying selection in endogenous retroviruses in the saltwater crocodile (Crocodylus porosus) in the Northern Territory of Australia.
Endogenous retroviruses (ERVs) are remnants of exogenous retroviruses that have integrated into the nuclear DNA of a germ-line cell. Here we present the results of a survey into the ERV complement of Crocodylus porosus, the saltwater crocodile, representing 45 individuals from 17 sampling locations in the Northern Territory of Australia. These retroelements were compared with published ERVs from other species of Crocodylia (Crocodilians; alligators, caimans, gharials and crocodiles) as well as representatives from other vertebrates. This study represents one of the first in-depth studies of ERVs within a single reptilian species shedding light on the diversity of ERVs and proliferation mechanisms in crocodilians. Analyses of the retroviral pro-pol gene region have corroborated the presence of two major clades of ERVs in C. porosus and revealed 18 potentially functional fragments out of the 227 recovered that encode intact pro-pol ORFs. Interestingly, we have identified some patterns of diversification among those ERVs as well as a novel sequence that suggests the presence of an additional retroviral genus in C. porosus. In addition, considerable diversity but low genetic divergence within one of the C. porosus ERV lineages was identified. We propose that the ERV complement of C. porosus has come about through a combination of recent infections and replication of ancestral ERVs. Strong purifying selection acting on these clades suggests that this activity is recent or still occurring in the genome of this species. The discovery of potentially functional elements is an interesting development that warrants further investigation.


Quantitative phylogenetic assessment of microbial communities in diverse environments.
The taxonomic composition of environmental communities is an important indicator of their ecology and function. We used a set of protein-coding marker genes, extracted from large-scale environmental shotgun sequencing data, to provide a more direct, quantitative, and accurate picture of community composition than that provided by traditional ribosomal RNA-based approaches depending on the polymerase chain reaction. Mapping marker genes from four diverse environmental data sets onto a reference species phylogeny shows that certain communities evolve faster than others. The method also enables determination of preferred habitats for entire microbial clades and provides evidence that such habitat preferences are often remarkably stable over time.


Echocardiographic deformation imaging reveals preserved regional systolic function in endurance athletes with left ventricular hypertrophy.
Left ventricular hypertrophy (LVH) is often observed in athletes, which should be differentiated from hypertrophic cardiomyopathy. The aim of the study was to explore the functional changes measured using tissue Doppler imaging (TDI) deformation analysis in athletes fulfilling LVH criteria participating in different endurance sports. Healthy controls (n = 62, 58% men) and endurance athletes (n = 120, 62% men) aged 18-40 years were prospectively enrolled and underwent both standard echocardiography as well as TDI. Longitudinal TDI-derived strain and strain rate (SR) were calculated in the septal and posterior wall in three segments. LVH was defined as a left ventricular mass (LVM) over 132 g/m2 in men and over 109 g/m2 in women. Echocardiographic LVH was observed in 33 athletes (67% men). LVM was significantly increased in both athlete groups (102.6 g/m2 (SD 16.0) and 135.7 g/m2 (SD 15.9) vs 88.0 g/m2 (SD 16.5) in controls, p<0.001). Diastolic parameters were not significantly different between groups. Athletes with LVH showed no significant difference in strain and SR values in any segment of the septal or posterior wall compared with controls or those without LVH. A weak but significant correlation (also after multivariate analysis) was found for septal wall thickness and LVM in peak systolic strain (r = 0.26, p<0.01 and 0.23, p<0.01) and SR (r = 0.27, p<0.01 and 0.29, p<0.01). Nevertheless, strain and SR values were still within normal limits in all athletes. Athletes with LVH overall show normal deformation values in the left ventricle. These data suggest that a moderate reduction in regional septal deformation should not be considered as pathological when evaluating the endurance athlete with echocardiographic LVH of unknown origin.


Bimanual training in stroke: How do coupling and symmetry-breaking matter?
The dramatic consequences of stroke on patient autonomy in daily living activities urged the need for new reliable therapeutic strategies. Recently, bimanual training has emerged as a promising tool to improve the functional recovery of upper-limbs in stroke patients. However, who could benefit from bimanual therapy and how it could be used as a part of a more complete rehabilitation protocol remain largely unknown. A possible reason explaining this situation is that coupling and symmetry-breaking mechanisms, two fundamental principles governing bimanual behaviour, have been largely under-explored in both research and rehabilitation in stroke. Bimanual coordination emerges as an active, task-specific assembling process where the limbs are constrained to act as a single unit by virtue of mutual coupling. Consequently, exploring, assessing, re-establishing and exploiting functional bimanual synergies following stroke, require moving beyond the classical characterization of performance of each limb in separate and isolated fashion, to study coupling signatures at both neural and behavioural levels. Grounded on the conceptual framework of the dynamic system approach to bimanual coordination, we debated on two main assumptions: 1) stroke-induced impairment of bimanual coordination might be anticipated/understood by comparing, in join protocols, changes in coupling strength and asymmetry of bimanual discrete movements observed in healthy people and those observed in stroke; 2) understanding/predicting behavioural manifestations of decrease in bimanual coupling strength and/or increase in interlimb asymmetry might constitute an operational prerequisite to adapt therapy and better target training at the specific needs of each patient. We believe that these statements draw new directions for experimental and clinical studies and contribute in promoting bimanual training as an efficient and adequate tool to facilitate the paretic upper-limb recovery and to restore spontaneous bimanual synergies. Since bimanual control deficits have scarcely been systematically investigated, the eventual benefits of bimanual coordination practice in stroke rehabilitation remains poorly understood. In the present paper we argued that a better understanding of coupling and symmetry-breaking mechanisms in both the undamaged and stroke-lesioned neuro-behavioral system should provide a better understanding of stroke-related alterations of bimanual synergies, and help clinicians to adapt therapy in order to maximize rehabilitation benefits.


Burning Velocity of Turbulent Methane/Air Premixed Flames in Subatmospheric Environments.
The aim of our work was to study turbulent premixed flames in subatmospheric conditions. For this purpose, turbulent premixed flames of lean methane/air mixtures were stabilized in a nozzle-type Bunsen burner and analyzed using Schlieren visualization and image processing to calculate turbulent burning velocities by the mean-angle method. Moreover, hot-wire anemometer measurements were performed to characterize the turbulent aspects of the flow. The environmental conditions were 0.85 atm, 0.98 atm, and 295 ± 2 K. The turbulence-flame interaction was analyzed based on the geometric parameters combined with laminar flame properties (which were experimentally and numerically determined), integral length scale, and Kolmogorov length scale. Our results show that the effects of subatmospheric pressure on turbulent burning velocity are significant. The ratio between turbulent and laminar burning velocities increases with turbulence intensity, but this effect tends to decrease as the atmospheric pressure is reduced. We propose a general empirical correlation as a function between S T/S L and u'/S L based on the experimental results obtained in this study and the equivalence ratio and pressure we established.


Advance care planning: identifying system-specific barriers and facilitators.
Advance care planning (acp) is an important process in health care today. How to prospectively identify potential local barriers and facilitators to uptake of acp across a complex, multi-sector, publicly funded health care system and how to develop specific mitigating strategies have not been well characterized. We surveyed a convenience sample of clinical and administrative health care opinion leaders across the province of Alberta to characterize system-specific barriers and facilitators to uptake of acp. The survey was based on published literature about the barriers to and facilitators of acp and on the Michie Theoretical Domains Framework. Of 88 surveys, 51 (58%) were returned. The survey identified system-specific barriers that could challenge uptake of acp. The factors were categorized into four main domains. Three examples of individual system-specific barriers were "insufficient public engagement and misunderstanding," "conflict among different provincial health service initiatives," and "lack of infrastructure." Local system-specific barriers and facilitators were subsequently explored through a semi-structured informal discussion group involving key informants. The group identified approaches to mitigate specific barriers. Uptake of acp is a priority for many health care systems, but bringing about change in multi-sector health care systems is complex. Identifying system-specific barriers and facilitators to the uptake of innovation are important elements of successful knowledge translation. We developed and successfully used a simple and inexpensive process to identify local system-specific barriers and enablers to uptake of acp, and to identify specific mitigating strategies.


K antigen and serum sensitivity of rough Escherichia coli.
We prepared bacterial hybrids which express both K1 and K27 antigens and examined the relative contributions of these capsules to serum resistance. Escherichia coli Hfr strain F639 (rough, K27+, serum sensitive) was conjugated with E. coli recipient strain E412 (rough, K1+, His- Trp- Strr, serum resistant). Transconjugants which inherited both the his and trp linked genes for K27 antigen synthesis were analyzed. These hybrids retained and expressed the K1 antigen since the K1 locus is nonallelic with K27 gene loci. Hybrid strains which express both K1 and K27 antigens exhibit serum resistance, but not at the level of the K1+ parental strain. An isogenic K1 derivative of a hybrid which expressed only K27 antigen was serum sensitive (greater than 99% kill, 60 min). These findings indicate that the presence of the K1 capsular antigen can protect some rough strains of E. coli from serum bactericidal activity, whereas K27 and perhaps other K antigens fail to provide such a protective effect.


On Power and Freedom: Extending the Definition of Coercion.
Defining coercion has been a topic of interest to behavior analysts from time to time. Given the more and more subtle influence strategies that technology has enabled, it is time to revisit these definitions. This article examines the definitions of power, freedom, and coercion in behavior analysis, comparing them to philosophical views of power, freedom, and coercion. Two extensions to the definition of coercion are suggested. First, definitions could include as coercive the removal of resources needed to generate the responses required to obtain reinforcement, or in some cases, the neglect to provide these resources. Second, choice architecture systems that are not transparent to the individuals being influenced and for which their consent has not been provided could be considered to be coercive. Implications of these extensions are discussed, including the need to examine behavior management methods for interactions considered to be coercive under the new definitions.


Trauma quality improvement: The Pietermaritzburg Metropolitan Trauma Service experience with the development of a comprehensive structure to facilitate quality improvement in rural trauma and acute care in KwaZulu-Natal, South Africa.
Improving the delivery of efficient and effective surgical care in rural South Africa is a mammoth task bedevilled by conflict between the stakeholders, who include rural doctors, surgeons, ancillary staff, researchers, educators and administrators. Management training is not part of most medical school curricula, yet as they progress in their careers, many clinicians are required to manage a health system and find the shift from caring for individual patients to managing a complex system difficult. Conflict arises when management-type interventions are imposed in a top-down manner on surgical staff suspicious of an unfamiliar field of study. Another area of conflict concerns the place of surgical research. Researchers are often accused of not being sufficiently focused on or concerned about the tasks of service delivery. This article provides an overview of management theory and describes a comprehensive management structure that integrates a model for health systems with a strategic planning process, strategic planning tools and appropriate quality metrics, and shows how the Pietermaritzburg Metropolitan Trauma Service in KwaZulu-Natal Province, South Africa, successfully used this structure to facilitate and contextualise a diverse number of quality improvement programmes and research initiatives in the realm of rural acute surgery and trauma. We have found this structure to be useful, and hope that it may be applied to other acute healthcare systems.


Single molecule high-resolution colocalization of Cy3 and Cy5 attached to macromolecules measures intramolecular distances through time.
Here we present a technique called single-molecule high-resolution colocalization (SHREC) of fluorescent dyes that allows the measurement of interfluorophore distances in macromolecules and macromolecular complexes with better than 10-nm resolution. By using two chromatically differing fluorescent molecules as probes, we are able to circumvent the Rayleigh criterion and measure distances much smaller than 250 nm. The probes are imaged separately and localized individually with high precision. The registration between the two imaging channels is measured by using fiduciary markers, and the centers of the two probes are mapped onto the same space. Multiple measurements can be made before the fluorophores photobleach, allowing intramolecular and intermolecular distances to be tracked through time. This technique's lower resolution limit lies at the upper resolution limit of single molecule FRET (smFRET) microscopy. The instrumentation and fluorophores used for SHREC can also be used for smFRET, allowing the two types of measurements to be made interchangeably, covering a wide range of interfluorophore distances. A dual-labeled duplex DNA molecule (30 bp) was used as a 10-nm molecular ruler to confirm the validity of the method. We also used SHREC to study the motion of myosin V. We directly observed myosin V's alternating heads while it walked hand-over-hand along an actin filament.


Phenoxy herbicides and soft-tissue sarcomas in female rice weeders. A population-based case-referent study.
A population-based case-referent study was conducted in an area of northern Italy where rice growing is the predominant agricultural activity and phenoxy herbicides have been used since 1950. Manual rice weeding was formerly performed by a seasonal female working population; in the early 1950s these women were concurrently exposed to chemical herbicides. Sixty-eight persons representing incident and histologically revised cases (31 women) and 158 population referents (73 women) were interviewed. The cases were histologically confirmed independently by two blinded pathologists, and exposure to phenoxy herbicides was assessed by two blinded pesticide researchers. An age-adjusted odds ratio of 0.91 was found for the living men (with suspect exposures; no man diagnosed as a case had been exposed with certainty to phenoxy herbicides). Among the living women the relative risk was 2.7 (90% confidence interval 0.59-12.37), and it further increased when attention was restricted to women exposed in the whole 1950-1955 period and to younger age groups.


Acetylcholinesterase responsive polymeric supra-amphiphiles for controlled self-assembly and disassembly.
We have fabricated enzyme responsive polymeric supra-amphiphiles by mixing a block copolymer of poly(ethylene glycol)-block-poly(acrylic acid) with myristoylcholine chloride in water. The polymeric supra-amphiphiles self-assemble into spherical aggregates with sizes varying from about 40 to 150 nm. Moreover, the spherical aggregates can be disassembled triggered by acetylcholinesterase, an enzyme which can cut off the ester linkage of myristoylcholine chloride. Nile red can be loaded into the spherical aggregates and released in several hours upon the treatment of acetylcholinesterase. The releasing rate is rather fast considering that it takes more than 150 h for Nile red to diffuse out of the spherical aggregates without addition of acetylcholinesterase. It is anticipated that the new enzyme responsive polymeric supra-amphiphile may be explored as a carrier for drug delivery.


Intimate partner violence and psychosocial health, a cross-sectional study in a pregnant population.
The objective of this paper is to explore whether IPV 12 months before and/or during pregnancy is associated with poor psychosocial health. From June 2010 to October 2012, a cross-sectional study was conducted in 11 antenatal care clinics in Belgium. Consenting pregnant women were asked to complete a questionnaire on socio-demographics, psychosocial health and violence in a separate room. Overall, 2586 women were invited to participate and we were able to use data from 1894 women (73.2%) for analysis. Ethical clearance was obtained in all participating hospitals. We found a significant correlation between IPV and poor psychosocial health: within the group of women who reported IPV, 53.2% (n = 118) had poor psychosocial health, as compared to 21% (n = 286) in the group of women who did not report IPV (P < 0.001). Lower psychosocial health scores were associated with increased odds of reporting IPV (aOR 1.55; 95% CI 1.39-1.72), with adjustments made for the language in which the questionnaire was filled out, civil/marital status, education and age. In other words, a decrease of 10 points on the psychosocial health scale (total of 140) increased the odds of reporting IPV by 55 %. When accounting for the 6 psychosocial health subscales, the analysis revealed that all subscales (depression, anxiety, self-esteem, mastery, worry and stress) are strongly correlated to reporting IPV. However, when accounting for all subscales simultaneously in a logistic regression model, only depression (aOR 0.87; 95 % CI 0.84-0.91) and stress (aOR 0.85; 95 % CI 0.77-095) remained significantly associated with IPV. The association between overall psychosocial health and IPV remained significant after adjusting for socio-demographic status. Our research corroborated that IPV and psychosocial health are strongly associated. Due to the limitations of our study design, we believe that future research is needed to deepen understanding of the multitude of factors involved in the complex interactions between IPV and psychosocial health.


Characterization of three sialidases from Danio rerio.
Zebrafish encodes several sialidases belonging to the NEU3 group, the plasma membrane-associated member of the family with high specificity toward ganglioside substrates. Neu3.1, Neu3.2 and Neu 3.3 have been expressed in E. coli and purified using the pGEX-2T expression system. Although all the enzymes are expressed by bacterial cells, Neu3.1 formed insoluble aggregates that hampered its purification. Neu3.2 and Neu3.3 formed oligomers as demonstrated by gel filtration chromatography experiments. Actually, the first formed a trimer whereas the second a pentamer. Intriguingly, despite relevant degree of sequence identity and similarity, the two enzymes showed peculiar substrate specificities toward gangliosides other than GM3, two glycoproteins and two forms of sialyllactose. Using molecular modelling and the crystal structure of the human cytosolic sialidase NEU2 as a template, the 3D models of the sialidases from zebrafish have been generated. As expected, the 3D models showed the typical six blade beta-propeller typical of sialidases, with an overall highly conserved active site architecture. The differences among the three zebrafish enzymes and human NEU2 are mainly located in the loops connecting the antiparallel beta strands of the propeller core. These portions of the proteins are probably responsible for the differences observed in substrate specificities, as well as in the different subcellular localization and aggregation features observed in solution. Finally, the in silico analysis of RNA-Seq data evidenced a peculiar expression profile of the three genes during embryogenesis, suggesting different roles of these sialidases during development.


Cytokine-induced expression of vascular cell adhesion molecule-1 (VCAM-1) by astrocytes and astrocytoma cell lines.
Astrocytes in the central nervous system (CNS) associate intimately with vascular endothelial cells and are integral to the blood-brain barrier (BBB). Leukocyte transmigration across the BBB is a cardinal feature of CNS inflammation, as observed in experimental autoimmune encephalomyelitis, and very late antigen-4 (VLA-4)/vascular cell adhesion molecule-1 (VCAM-1) interactions have recently been proposed as essential for this process. VCAM-1 expression by astrocytes was recently reported. We addressed the regulation of VCAM-1 expression by inflammatory cytokines in primary human astrocytes and two human astrocytoma cell lines. Astrocytoma cells up-regulated surface VCAM-1 expression in response to cytokines in the following order: IFN-gamma plus TNF-alpha > IFN-gamma plus IL-1 beta > TNF-alpha > IFN-gamma. Cytokine-activated astrocytoma cells expressed 7-domain VCAM-1, as indicated by accumulation of 3.2-kb VCAM-1 mRNA and immunoprecipitation of a 100-kDa protein with anti-VCAM-1 mAb. Lymphoblast adhesion to cytokine-activated astrocytoma cell monolayers was significantly blocked by mAb specific for VCAM-1 and VLA-4, indicating that astrocytoma cell VCAM-1 was functional. Astrocytoma cell expression of VCAM-1 could be a constituent of the astrocyte phenotype. To support this possibility, we demonstrated that cytokine-treated adult human and rat primary astrocytes expressed VCAM-1, and the rank order of cytokine potency for VCAM-1 induction in primary and neoplastic astrocytes was strikingly similar. This is the first documentation of VCAM-1 expression by adult human astrocytes. Expression of VCAM-1 by astrocytes at the BBB could play a role in mononuclear leukocyte entry into the CNS.


Hemodynamic mechanisms of the attenuated blood pressure response to mental stress after a single bout of maximal dynamic exercise in healthy subjects.
To determine the hemodynamic mechanisms responsible for the attenuated blood pressure response to mental stress after exercise, 26 healthy sedentary individuals (age 29 ± 8 years) underwent the Stroop color-word test before and 60 min after a bout of maximal dynamic exercise on a treadmill. A subgroup (N = 11) underwent a time-control experiment without exercise. Blood pressure was continuously and noninvasively recorded by infrared finger photoplethysmography. Stroke volume was derived from pressure signals, and cardiac output and peripheral vascular resistance were calculated. Perceived mental stress scores were comparable between mental stress tests both in the exercise (P = 0.96) and control (P = 0.24) experiments. After exercise, the blood pressure response to mental stress was attenuated (pre: 10 ± 13 vs post: 6 ± 7 mmHg; P < 0.01) along with lower values of systolic blood pressure (pre: 129 ± 3 vs post: 125 ± 3 mmHg; P < 0.05), stroke volume (pre: 89.4 ± 3.5 vs post: 76.8 ± 3.8 mL; P < 0.05), and cardiac output (pre: 7.00 ± 0.30 vs post: 6.51 ± 0.36 L/min; P < 0.05). Except for heart rate, the hemodynamic responses and the mean values during the two mental stress tests in the control experiment were similar (P > 0.05). In conclusion, a single bout of maximal dynamic exercise attenuates the blood pressure response to mental stress in healthy subjects, along with lower stroke volume and cardiac output, denoting an acute modulatory action of exercise on the central hemodynamic response to mental stress.


Body Weight Variability Increases Dementia Risk Among Older Adults: A Nationwide Population-Based Cohort Study.
Background: Recent growing evidences suggest that body weight (Bwt) variability, a repeated loss and regain of weight within a specific period, causes metabolic disturbances and can be a marker for poor homeostasis. Although there have been many studies about the association between Bwt variability and various health status, its association with the incidence of dementia among elderly people has not been examined. Methods: We performed a retrospective elderly cohort study from 19,987 participants with mean age 73 years old in the Korean National Health Insurance Service. We examined the risk of incident dementia, including Alzheimer's dementia and vascular dementia, according to the quartile of Bwt variability, represented as coefficient of variation (Bwt-CV), SD (Bwt-SD), and variability independent of the mean (Bwt-VIM). Results: In fully adjusted model, the group with the highest Bwt variability (Bwt-VIM Q4) showed an increased risk of all-cause dementia (hazard ratio [HR] 1.39, 95% confidence interval [CI] 1.206-1.603) and Alzheimer's dementia (HR 1.46, CI 1.240-1.724) compared to the lowest quartile (Bwt-VIM Q1). We also found that subjects with the highest Bwt variability (Q4) and underweight BMI had a significantly increased risk of developing dementia (HR 1.994, 95% CI 1.302-3.054), while subjects with low Bwt variability (Q1 and Q2) and obese BMI had decreased risk of dementia (HR 0.664, 95% CI 0.505-0.872 and HR 0.648, 95% CI 0.493-0.852, respectively) compared to reference group (lowest Bwt variability (Q1) with normal baseline BMI). The effect of Bwt variability on the incidence of dementia was more prominent in subjects <75 years old and abnormal BMI groups (P for interaction < 0.05). Conclusion: The present study revealed that high Bwt variability was associated with an increased risk of dementia in the elderly.


The CD3 zeta cytoplasmic domain mediates CD2-induced T cell activation.
CD2-mediated T lymphocyte activation requires surface expression of CD3-Ti, the T cell receptor (TCR) for antigen major histocompatibility complex protein. Given the importance of CD3 zeta in TCR signaling, we have directly examined the ability of the CD3 zeta cytoplasmic domain to couple CD2 to intracellular signal transduction pathways. A cDNA encoding a chimeric protein consisting of the human CD3 zeta cytoplasmic domain (amino acid residues 31-142) fused to the CD8 alpha extracellular and transmembrane domains (amino acid residues 1-187) was transfected into a CD2+CD3-CD8- variant of the human T cell line Jurkat. The resulting transfectants expressed the CD8 alpha/CD3 zeta chimeric receptor at the cell surface in the absence of other TCR subunits. Stimulation of these transfectants with anti-T11(2) + anti-T11(3) monoclonal antibodies (mAbs) initiated both a prompt cytosolic free calcium ([Ca2+]i) rise and protein tyrosine kinase activation. Stimulation with either intact anti-T11(2) + anti-T11(3) mAbs or purified F(ab')2 fragments resulted in interleukin 2 (IL-2) secretion. In contrast, control cell lines transfected with a cDNA encoding wild-type CD8 alpha, and thus lacking surface expression of the CD3 zeta cytoplasmic domain, failed to show any [Ca2+]i rise, protein tyrosine kinase activation, or IL-2 secretion after identical stimulation. These data directly establish the CD3 zeta cytoplasmic domain as a necessary and sufficient component of the CD3-Ti complex involved in T lymphocyte activation through CD2. Moreover, they show that CD2 signaling can function in the absence of Fc receptors.


Skin diseases in companion guinea pigs (Cavia porcellus): a retrospective study of 293 cases seen at the Veterinary Medical Teaching Hospital, University of California at Davis (1990-2015).
Guinea pigs (Cavia porcellus) are popular companion animals with reported skin diseases, but most reports are accessed from textbooks or review articles. To document skin diseases and their prevalence in companion guinea pigs in northern California, USA, and to investigate predilections for the most common conditions over a 25 year period. Two hundred and ninety three guinea pigs from the hospital population met inclusion criteria. A retrospective study was performed by searching computerized medical records seen at the Veterinary Medical Teaching Hospital (VMTH), School of Veterinary Medicine, University of California from 1 January 1990 to 31 July 2015 using key words relevant to dermatology. Of the 580 guinea pigs seen at the VMTH, 50% (293) had skin disease. Of the 293 cases, 154 (52%) presented for nondermatological reasons. Guinea pigs with skin disease were significantly older than those without skin disease (P = 0.0002); females with skin disease were more likely to have cystic ovaries (P = 0.0203), although these were not always associated with alopecia. Pododermatitis and infestation with Trixacarus caviae or lice were the most common skin diseases. Ivermectin or selamectin was used for ectoparasite treatment. Abscesses unassociated with pododermatitis were the most common nodules. Benign follicular tumours were the most common neoplasm. Despite the frequent mention of dermatophytosis in the veterinary literature, only two cases of Trichophyton mentagrophytes were diagnosed. Cutaneous conditions in companion guinea pigs in the USA are common. Clinicians should include a dermatological examination when examining these rodents regardless of the reason for presentation.


Topiramate plus Cooling for Hypoxic-Ischemic Encephalopathy: A Randomized, Controlled, Multicenter, Double-Blinded Trial.
Therapeutic interventions to improve the efficacy of whole-body cooling for hypoxic-ischemic encephalopathy (HIE) are desirable. Topiramate has been effective in reducing brain damage in experimental studies. However, in the clinical setting information is limited to a small number of feasibility trials. We launched a randomized controlled double-blinded topiramate/placebo multicenter trial with the primary objective being to reduce the antiepileptic activity in cooled neonates with HIE and assess if brain damage would be reduced as a consequence. Neonates were randomly assigned to topiramate or placebo at the initiation of hypothermia. Topiramate was administered via a nasogastric tube. Brain electric activity was continuously monitored. Topiramate pharmacokinetics, energy-related and Krebs' cycle intermediates, and lipid peroxidation biomarkers were determined using liquid chromatography-mass spectrometry and MRI for assessing brain damage. Out of 180 eligible patients 110 were randomized, 57 (51.8%) to topiramate and 53 (48.2%) to placebo. No differences in the perinatal or postnatal variables were found. The topiramate group exhibited less seizure burden in the first 24 h of hypothermia (topiramate, n = 14 [25.9%] vs. placebo, n = 22 [42%]); needed less additional medication, and had lower mortality (topiramate, n = 5 [9.2%] vs. placebo, n = 10 [19.2%]); however, these results did not achieve statistical significance. Topiramate achieved a therapeutic range in 37.5 and 75.5% of the patients at 24 and 48 h, respectively. A significant association between serum topiramate levels and seizure activity (p < 0.016) was established. No differences for oxidative stress, energy-related metabolites, or MRI were found. Topiramate reduced seizures in patients achieving therapeutic levels in the first hours after treatment initiation; however, they represented only a part of the study population. Our results warrant further studies with higher loading and maintenance dosing of topiramate.


Experimental Campylobacter jejuni infection in the chicken: an animal model of axonal Guillain-Barré syndrome.
To develop and characterise an animal model of paralytic neuropathy after Campylobacter jejuni infection. Campylobacter infection precedes development of many cases of Guillain-Barré syndrome and is particularly associated with cases having prominent axonal degeneration. Understanding the pathogenesis of Guillain-Barré syndrome after C jejuni infection has been slowed by the lack of animal models. A spontaneous paralytic neuropathy is described that developed in chickens from the farms of four patients with Guillain-Barré syndrome. The production of paralytic neuropathy in chickens experimentally fed Campylobacter jejuni isolated from one of these patients is reported. The sciatic nerves of the spontaneously paralysed chickens were examined pathologically in teased fibres, in plastic embedded sections, and by electron microscopy. Two large groups of chickens were then fed cultures of a C jejuni (Penner type O:19) isolated from one of these patients. The chickens with spontaneous paralysis had pathologically noninflammatory neuropathy. Pathology in the sciatic nerves ranged from no detectable changes to severe Wallerian-like degeneration. In the experimentally inoculated groups, an average of 33% of the chickens became paralysed. The median time after inoculation to paralysis was 12 days. The lesions found in the first few days of paralysis included nodal lengthening and paranodal demyelination. In those animals that survived for several days after onset of weakness, the pathology was dominated by extensive Wallerian-like degeneration. Animals that survived for weeks with no clinically apparent neuropathy had paranodal remyelination in some teased nerve fibres, reflecting earlier paranodal demyelination. Experimental inoculation with C jejuni may provide a new model for understanding some forms of Guillain-Barré syndrome.


Prediction of Peromyscus maniculatus (deer mouse) population dynamics in Montana, USA, using satellite-driven vegetation productivity and weather data.
Deer mice (Peromyscus maniculatus) are the main reservoir host for Sin Nombre virus, the primary etiologic agent of hantavirus pulmonary syndrome in North America. Sequential changes in weather and plant productivity (trophic cascades) have been noted as likely catalysts of deer mouse population irruptions, and monitoring and modeling of these phenomena may allow for development of early-warning systems for disease risk. Relationships among weather variables, satellite-derived vegetation productivity, and deer mouse populations were examined for a grassland site east of the Continental Divide and a sage-steppe site west of the Continental Divide in Montana, USA. We acquired monthly deer mouse population data for mid-1994 through 2007 from long-term study sites maintained for monitoring changes in hantavirus reservoir populations, and we compared these with monthly bioclimatology data from the same period and gross primary productivity data from the Moderate Resolution Imaging Spectroradiometer sensor for 2000-06. We used the Random Forests statistical learning technique to fit a series of predictive models based on temperature, precipitation, and vegetation productivity variables. Although we attempted several iterations of models, including incorporating lag effects and classifying rodent density by seasonal thresholds, our results showed no ability to predict rodent populations using vegetation productivity or weather data. We concluded that trophic cascade connections to rodent population levels may be weaker than originally supposed, may be specific to only certain climatic regions, or may not be detectable using remotely sensed vegetation productivity measures, although weather patterns and vegetation dynamics were positively correlated.


Serum urate during acute gout.
To study the frequency of normal serum urate (SU) levels during acute gout in the largest studies of acute gout treatment to date. Data collected from 2 randomized controlled clinical trials assessing the efficacy of etoricoxib or indomethacin for 7 days in acute gout were used to assess SU levels during acute gouty attacks. Efficacy was similar with both agents, so both groups were combined for analysis. A total of 339 patients were enrolled in the 2 studies; 94% were male; mean age was 50.5 years. At baseline, 14% of patients had a "true" normal SU (<or=6 mg/dl) and 32% had SU<or=8 mg/dl during acute gout. Baseline mean SU was 7.1 versus 8.5 mg/dl (p<0.001) in those taking allopurinol versus nonusers. Patients taking chronic allopurinol were more likely to have lower SU at baseline compared to those not taking chronic allopurinol (p<0.001) during the acute attack. A normal SU level at presentation does not exclude an acute gouty attack. In the largest studies of acute gout to date, attacks still occurred despite SU levels being below 6.8 mg/dl, the saturation level for urate. This may be attributed to persistence of tophi and an increased body uric acid pool. Additional studies are needed to determine the correlation between SU and the body uric acid pool as well as the relationship to timing of changes during acute gout.


Epidural spinal-cord stimulation facilitates recovery of functional walking following incomplete spinal-cord injury.
We investigated a novel treatment paradigm for developing functional ambulation in wheelchair-dependent individuals with chronic, incomplete spinal-cord injury. By coordinating epidural stimulation of the dorsal structures of the spinal cord with partial weight bearing treadmill therapy, we observed improvement in treadmill and over-ground ambulation in an individual with chronic incomplete tetraplegia. The application of partial weight-bearing therapy alone was not sufficient to achieve functional ambulation over ground, though treadmill ambulation improved significantly. Combining epidural spinal-cord stimulation (ESCS, T10-T12 vertebral levels) with partial weight-bearing therapy resulted in further improvement during treadmill ambulation. Moreover, the combination of therapies facilitated the transfer of the learned gait into over ground ambulation. Performance improvements were elicited by applying continuous, charge-balanced, monophasic pulse trains at a frequency of 40-60 Hz, a pulse duration of 800 micros, and an amplitude determined by the midpoint (50%) between the sensory and motor threshold values. The participant initially reported a reduction in sense of effort for over ground walking from 8/10 to 3/10 (Borg scale), and was able to double his walking speed. After several weeks of over ground training, he reached maximum walking speeds of 0.35 m/s, and was able to ambulate over 325 m. We propose that ESCS facilitated locomotor recovery in this patient by augmenting the use-dependent plasticity created by partial weight bearing therapy. Confirmation of these promising results in a controlled study of groups of spinal-cord-injured subjects is warranted.


Deciphering the aqueous chemistry of glyoxal oxidation with hydrogen peroxide using molecular imaging.
Aqueous surfaces after photochemical and dark reactions of glyoxal and hydrogen peroxide (H2O2) have been studied using a microfluidic reactor coupled with an in situ liquid time-of-flight secondary ion mass spectrometry (ToF-SIMS) for the first time. Spectral principal component analysis was used to determine similarities and differences among various photochemical aging and dark reaction samples and controls. Compared with previous results using bulk solutions, our unique liquid surface molecular imaging approach provided observations of glyoxal hydration (i.e., first and secondary products), oxidation products (i.e., glyoxylic acid, oxalic acid, formic acid, tartaric acid), oligomers, cluster ions, and water clusters with sub-micrometer spatial resolution. Observations of oxidation products give the physical foundation to deduce new reaction pathways at the aqueous surface. The first chemical mapping of water cluster changes between dark and photochemical aging suggests that glyoxal oxidation affects the hydrophobicity and water microenvironment at the surface, influencing the particle's ability of reactive uptake and subsequent cloud condensation nucleation and/or ice nucleation activation. Moreover, SIMS three-dimensional chemical mapping has made it possible to visualize the surface mixing state for the first time. We potentially provide a new method to investigate complex surface chemistry as an important source of aqueous secondary organic aerosol (aqSOA) formation in atmospheric chemistry.


Prevalence of veterinary antibiotics and antibiotic-resistant Escherichia coli in the surface water of a livestock production region in northern China.
This study investigated the occurrence of 12 veterinary antibiotics (VAs) and the susceptibility of Escherichia coli (E. coli) in a rural water system that was affected by livestock production in northern China. Each of the surveyed sites was determined with at least eight antibiotics with maximum concentration of up to 450 ng L(-1). The use of VAs in livestock farming probably was a primary source of antibiotics in the rivers. Increasing total antibiotics were measured from up- to mid- and downstream in the two tributaries. Eighty-eight percent of the 218 E. coli isolates that were derived from the study area exhibited, in total, 48 resistance profiles against the eight examined drugs. Significant correlations were found among the resistance rates of sulfamethoxazole-trimethoprim, chloromycetin and ampicillin as well as between tetracycline and chlortetracycline, suggesting a possible cross-selection for resistance among these drugs. The E. coli resistance frequency also increased from up- to midstream in the three rivers. E. coli isolates from different water systems showed varying drug numbers of resistance. No clear relationship was observed in the antibiotic resistance frequency with corresponding antibiotic concentration, indicating that the antibiotic resistance for E. coli in the aquatic environment might be affected by factors besides antibiotics. High numbers of resistant E. coli were also isolated from the conserved reservoir. These results suggest that rural surface water may become a large pool of VAs and resistant bacteria. This study contributes to current information on VAs and resistant bacteria contamination in aquatic environments particularly in areas under intensive agriculture. Moreover, this study indicates an urgent need to monitor the use of VAs in animal production, and to control the release of animal-originated antibiotics into the environment.


Sleep-dependent motor memory consolidation in older adults depends on task demands.
It is often suggested that sleep-dependent consolidation of motor learning is impaired in older adults. The current study challenges this view and suggests that the degree of motor consolidation seen with sleep in older age groups depends on the kinematic demands of the task. We show that, when tested with a classic sequence learning task, requiring individuated finger movements, older adults did not show sleep-dependent consolidation. By contrast, when tested with an adapted sequence learning task, in which movements were performed with the whole hand, sleep-dependent motor improvement was observed in older adults. We suggest that age-related decline in fine motor dexterity may in part be responsible for the previously described deficit in sleep-dependent motor consolidation with aging.


Characterisation of ethylene pathway components in non-climacteric capsicum.
Climacteric fruit exhibit high ethylene and respiration levels during ripening but these levels are limited in non-climacteric fruit. Even though capsicum is in the same family as the well-characterised climacteric tomato (Solanaceae), it is non-climacteric and does not ripen normally in response to ethylene or if harvested when mature green. However, ripening progresses normally in capsicum fruit when they are harvested during or after what is called the 'Breaker stage'. Whether ethylene, and components of the ethylene pathway such as 1-aminocyclopropane 1-carboxylate (ACC) oxidase (ACO), ACC synthase (ACS) and the ethylene receptor (ETR), contribute to non-climacteric ripening in capsicum has not been studied in detail. To elucidate the behaviour of ethylene pathway components in capsicum during ripening, further analysis is therefore needed. The effects of ethylene or inhibitors of ethylene perception, such as 1-methylcyclopropene, on capsicum fruit ripening and the ethylene pathway components may also shed some light on the role of ethylene in non-climacteric ripening. The expression of several isoforms of ACO, ACS and ETR were limited during capsicum ripening except one ACO isoform (CaACO4). ACS activity and ACC content were also low in capsicum despite the increase in ACO activity during the onset of ripening. Ethylene did not stimulate capsicum ripening but 1-methylcyclopropene treatment delayed the ripening of Breaker-harvested fruit. Some of the ACO, ACS and ETR isoforms were also differentially expressed upon treatment with ethylene or 1-methylcyclopropene. ACS activity may be the rate limiting step in the ethylene pathway of capsicum which restricts ACC content. The differential expression of several ethylene pathway components during ripening and upon ethylene or 1-methylclopropene treatment suggests that the ethylene pathway may be regulated differently in non-climacteric capsicum compared to the climacteric tomato. Ethylene independent pathways may also exist in non-climacteric ripening as evidenced by the up-regulation of CaACO4 during ripening onset despite being negatively regulated by ethylene exposure. However, some level of ethylene perception may still be needed to induce ripening especially during the Breaker stage. A model of capsicum ripening is also presented to illustrate the probable role of ethylene in this non-climacteric fruit.


Outcomes of Induction Therapy with Rabbit Anti-Thymocyte Globulin in Heart Transplant Recipients: A Single Center Retrospective Cohort Study.
BACKGROUND Induction immunosuppression is used in transplantation to prevent early acute rejection. The survival benefit of rabbit anti-thymocyte globulin (rATG) induction has not been established yet. We sought to determine the role of rATG in preventing rejection and improving overall survival. MATERIAL AND METHODS A retrospective cohort study was conducted from 2005 to 2009 and data of consecutive 268 heart transplant recipients were reviewed. RESULTS The data of 144 patients who received induction with rATG were compared to 124 patients who did not. Although overall survival was not different between the 2 groups (P=0.12), there was a significant difference in restricted mean survival time (RMST) at 5 years (RMST=4.8 months; 95% CI: 1.0-8.6, P=0.01) and 10 years (RMST=10.4 months; 95% CI: 1.6-19.3, P=0.02) in favor of the non-induced patients. No difference was observed between induced and non-induced patients who developed de novo donor specific antibodies. There was a significant difference in median days to first rejection in favor of the induced group (P<0.001). CONCLUSIONS Induction with rATG adds no survival benefit in heart transplant recipients. Patients who did not receive induction therapy had higher life expectancy at 5 years and 10 years. Although there was significant delay in the first rejection episode in favor of the rATG induced group, no difference was observed in donor specific antibodies. This study indicates a need for separate analysis of peri-transplantation co-morbidities and mainly the incidence of acute kidney injury, which could affect long-term survival.


Unique characteristics that distinguish suicide attempters from patients with nonsuicidal self-injury admitted to the emergency department following self-harm behavior: Psychological scales and biochemical markers.
The objective was to investigate the characteristics of suicide attempters as distinguished from nonsuicidal self-injury (NSSI) among those who are admitted to the emergency department (ED) following self-harm behavior using psychological scales and biochemical markers. The initial assessment forms and medical records of patients referred to the ED after self-harm behavior between March 2017 and December 2019 were retrospectively reviewed. Based on the patients' statements, two groups were formed: the NSSI group and the suicide attempt (SA) group. This study included 578 patients, 76.6% (n = 443) in the SA group and 23.4% (n = 135) in the NSSI group. A univariate analysis comparing the SA and NSSI groups revealed that the SA group was statistically significantly associated with male sex, chronic disease, history of depression, unemployment, not seeking help after the attempt, reduced consciousness, and psychiatric consultation. Further, the SA group was older and showed higher Risk-Rescue Rating Scale (RRRS) value and white blood cell value compared to the NSSI group. However, multivariate logistic analysis did not produce statistically significant results for RRRS and any of the biological markers, with one exception for the accessibility to rescue item in the RRRS, where the NSSI group showed a statistically higher "ask for help" score compared to the SA group (P < 0.01). Among patients who were admitted to the ED following self-harm behaviors, the risk factors for SA as opposed to NSSI were chronic disease and not seeking help after the attempt. In addition, suicide attempters tended to be more cooperative in psychiatric consultation following ED admission. In clinical practice, patients admitted due to NSSI should be administered treatment equivalent to that for suicide attempters if they exhibit the characteristics of suicide attempters.


Random monoallelic expression of three genes clustered within 60 kb of mouse t complex genomic DNA.
Mammals achieve gene dosage control by (1) random X-chromosome inactivation in females, (2) parental origin-specific imprinting of selected autosomal genes, and (3) random autosomal inactivation. Genes belonging to the third category of epigenetic phenomenon are just now emerging, with only six identified so far. Here we report three additional genes, Nubp2, Igfals, and Jsap1, that show 50%-methylated CpG sites by Southern blot analyses and primarily monoallelic expression in single-cell allele-specific RT-PCR analysis of bone marrow stromal cells and hepatocytes. Furthermore, we show that, in contrast to X inactivation, alleles can switch between active and inactive states during the formation of daughter cells. These three genes are the first in their category to exist as a tight cluster, in the proximal region of mouse chromosome 17, providing a thus far unique example of a region of autosomal random monoallelic expression.


Improvement in insulin sensitivity after human islet transplantation for type 1 diabetes.
Islet transplantation can improve metabolic control for type 1 diabetes (T1D), an effect anticipated to improve insulin sensitivity. However, current immunosuppression regimens containing tacrolimus and sirolimus have been shown to induce insulin resistance in rodents. The objective of the study was to evaluate the effect of islet transplantation on insulin sensitivity in T1D using euglycemic clamps with the isotopic dilution method to distinguish between effects at the liver and skeletal muscle. Twelve T1D subjects underwent evaluation in the Clinical and Translational Research Center before and between 6 and 7 months after the transplant and were compared with normal control subjects. The intervention included intrahepatic islet transplantation according to a Clinical Islet Transplantation Consortium protocol under low-dose tacrolimus and sirolimus immunosuppression. Total body (M/Δinsulin), hepatic (1/endogenous glucose production ·basal insulin) and peripheral [(Rd - endogenous glucose production)/Δinsulin] insulin sensitivity assessed by hyperinsulinemic (1 mU·kg(-1)·min(-1)) euglycemic (∼90 mg/dL) clamps with 6,6-(2)H2-glucose tracer infusion were measured. Glycosylated hemoglobin was reduced in the transplant recipients from 7.0% ± 0.3% to 5.6% ± 0.1% (P < .01). There were increases in total (0.11 ± 0.01 to 0.15 ± 0.02 dL/min·kg per microunit per milliliter), hepatic [2.3 ± 0.1 to 3.7 ± 0.4 × 10(2) ([milligrams per kilogram per minute](-1)·(microunits per milliliter)(-1))], and peripheral (0.08 ± 0.01 to 0.12 ± 0.02 dL/min·kg per microunit per milliliter) insulin sensitivity from before to after transplantation (P < .05 for all). All insulin sensitivity measures were less than normal in T1D before (P ≤ .05) and not different from normal after transplantation. Islet transplantation results in improved insulin sensitivity mediated by effects at both the liver and skeletal muscle. Modern dosing of glucocorticoid-free immunosuppression with low-dose tacrolimus and sirolimus does not induce insulin resistance in this population.


Bottom-up photonic crystal cavities formed by patterned III-V nanopillars.
We report on the formation and optical properties of bottom-up photonic crystal (PC) cavities formed by III-V nanopillars (NPs) via catalyst-free selective-area metal-organic chemical vapor deposition on masked GaAs substrates. This method of NP synthesis allows for precise lithographic control of NP position and diameter enabling simultaneous formation of both the photonic band gap (PBG) region and active gain region. The PBG and cavity resonance are determined by independently tuning the NP radius r, pitch a, and height h in the respective masked areas. Near-infrared emission at 970 nm is achieved from axial GaAs/InGaAs heterostructures with in situ passivation by laterally grown InGaP shells. To achieve out-of-plane optical confinement, the PC cavities are embedded in polydimethylsiloxane (PDMS) and removed from the growth substrate. Spatially and spectrally resolved 77 K photoluminescence demonstrates a strong influence of the PBG resonance on device emission. Resonant peaks are observed in the emission spectra of PC cavities embedded in PDMS.


Structural and biophysical insights into the mode of covalent binding of rationally designed potent BMX inhibitors.
The bone marrow tyrosine kinase in chromosome X (BMX) is pursued as a drug target because of its role in various pathophysiological processes. We designed BMX covalent inhibitors with single-digit nanomolar potency with unexploited topological pharmacophore patterns. Importantly, we reveal the first X-ray crystal structure of covalently inhibited BMX at Cys496, which displays key interactions with Lys445, responsible for hampering ATP catalysis and the DFG-out-like motif, typical of an inactive conformation. Molecular dynamic simulations also showed this interaction for two ligand/BMX complexes. Kinome selectivity profiling showed that the most potent compound is the strongest binder, displays intracellular target engagement in BMX-transfected cells with two-digit nanomolar inhibitory potency, and leads to BMX degradation PC3 in cells. The new inhibitors displayed anti-proliferative effects in androgen-receptor positive prostate cancer cells that where further increased when combined with known inhibitors of related signaling pathways, such as PI3K, AKT and Androgen Receptor. We expect these findings to guide development of new selective BMX therapeutic approaches.


Prevalence of neutralizing antibodies against California and Bunyamwera serogroup viruses in deer from mountainous areas of California.
Plaque reduction-serum dilution neutralization was used to evaluate the status of bunyavirus activity in deer in mountainous areas of California. Antibodies against 9 bunyaviruses were measured in 337 mule deer (Odocoileus hemionus hemionus, O. hemionus californicus, and O. hemionus inyoensis) and black-tailed deer (O. hemionus columbianus). More deer from high mountainous areas had neutralizing antibodies against Jamestown Canyon virus than did deer from low mountainous areas (23% vs. 9%; P less than 0.01). This finding is consistent with transmission by snow pool Aedes mosquitoes. Results for Jerry Slough virus were nearly identical to those for Jamestown Canyon virus, which is further evidence that these are strains of the same virus. Neutralizing antibodies against Northway virus were present in 26% of deer from high mountainous areas and 23% of deer from low mountainous areas, suggesting the involvement of a widespread vector, such as Culiseta inornata. Northway virus is not known to occur outside of Alaska and northwestern Canada. Low prevalences of antibodies were detected in deer to California encephalitis, La Crosse, and snowshoe hare viruses of the California serogroup; and Cache Valley, Lokern, and Main Drain viruses of the Bunyamwera serogroup.


Large oncosomes contain distinct protein cargo and represent a separate functional class of tumor-derived extracellular vesicles.
Large oncosomes (LO) are atypically large (1-10 µm diameter) cancer-derived extracellular vesicles (EVs), originating from the shedding of membrane blebs and associated with advanced disease. We report that 25% of the proteins, identified by a quantitative proteomics analysis, are differentially represented in large and nano-sized EVs from prostate cancer cells. Proteins enriched in large EVs included enzymes involved in glucose, glutamine and amino acid metabolism, all metabolic processes relevant to cancer. Glutamine metabolism was altered in cancer cells exposed to large EVs, an effect that was not observed upon treatment with exosomes. Large EVs exhibited discrete buoyant densities in iodixanol (OptiPrep(TM)) gradients. Fluorescent microscopy of large EVs revealed an appearance consistent with LO morphology, indicating that these structures can be categorized as LO. Among the proteins enriched in LO, cytokeratin 18 (CK18) was one of the most abundant (within the top 5th percentile) and was used to develop an assay to detect LO in the circulation and tissues of mice and patients with prostate cancer. These observations indicate that LO represent a discrete EV type that may play a distinct role in tumor progression and that may be a source of cancer-specific markers.


Stroke epidemiology: a review of population-based studies of incidence, prevalence, and case-fatality in the late 20th century.
This overview of population-based studies of incidence, prevalence, mortality, and case-fatality of stroke was based on studies from 1990. Incidence (first stroke in an individual's lifetime) and prevalence were computed by age, sex, and stroke type. Age-standardised incidence and prevalence with the corresponding 95% CI were plotted for each study to facilitate comparisons. The review shows that the burden of stroke is high and is likely to increase in future decades as a result of demographic and epidemiological transitions in populations. The main features of stroke epidemiology include modest geographical variation in incidence, prevalence, and case-fatality among the--predominantly white--populations studied so far, and a stabilisation or reversal in the declining secular trends in the pre-1990s rates, especially in older people. However, further research that uses the best possible methods to study the incidence, risk factors, and outcome of stroke are urgently needed in other populations of the world, especially in less developed countries where the risk of stroke is high, lifestyles are changing rapidly, and population restructuring is occurring.


Y27, a novel derivative of 4-hydroxyquinoline-3-formamide, prevents the development of murine systemic lupus erythematosus-like diseases in MRL/lpr autoimmune mice and BDF1 hybrid mice.
Naturally occurring CD4+CD25+ regulatory T (Treg) cells are central to the maintenance of peripheral tolerance. Impaired activity and/or a lower frequency of these cells lead to systemic lupus erythematosus (SLE). Manipulating the number or activity of Treg cells is to be a promising strategy in treating it and other autoimmune diseases. We have examined the effects of Y27, a novel derivative of 4-hydroxyquinoline-3-formamide, on SLE-like symptoms in MRL/lpr autoimmune mice and BDF1 hybrid mice. Whether the beneficial effect of Y27 involves modulation of CD4+CD25+ Treg cells has also been investigated. Female MRL/lpr mice that spontaneously develop lupus were treated orally by gavage with Y27 for 10 weeks, starting at 10 weeks of age. BDF1 mice developed a chronic graft-versus-host disease (GVHD) by two weekly intravenous injections of parental female DBA/2 splenic lymphocytes, characterized by immunocomplex-mediated glomerulonephritis resembling SLE. Y27 was administered to chronic GVHD mice for 12 weeks. Nephritic symptoms were monitored and the percentage of CD4+CD25+FoxP3+ Treg peripheral blood leukocyte was detected with mouse regulatory T cell staining kit by flowcytometry. Purified CD4+CD25+ Tregs were assessed for immune suppressive activity using the mixed lymphocyte reaction. The life-span of MRL/lpr mice treated with Y27 for 10 weeks was significantly prolonged, proteinuria and renal lesion severity were ameliorated, and blood urea nitrogen, triglyceride and serum anti-double-stranded DNA antibodies were decreased. Similar results were found in chronic GVHD mice. Administration of Y27 had little impact on percentage of the peripheral blood lymphocyte CD4+CD25+Foxp3+ Treg cells in both groups of mice. In contrast, the suppressive capacity of CD4+CD25+ Treg cells in splenocytes was markedly augmented in Y27-treated mice ex vivo. Experimental evidence of the protect effects of Y27 against autoimmune nephritis has been shown. The mechanism may involve enhancement of the suppressive capacity of CD4+CD25+ Treg cells.


Intraoperative neurophysiological mapping and monitoring in spinal tumor surgery: sirens or indispensable tools?
Spinal tumor (ST) surgery carries the risk of new neurological deficits in the postoperative period. Intraoperative neurophysiological monitoring and mapping (IONM) represents an effective method of identifying and monitoring in real time the functional integrity of both the spinal cord (SC) and the nerve roots (NRs). Despite consensus favoring the use of IONM in ST surgery, in this era of evidence-based medicine, there is still a need to demonstrate the effective role of IONM in ST surgery in achieving an oncological cure, optimizing patient safety, and considering medicolegal aspects. Thus, neurosurgeons are asked to establish which techniques are considered indispensable. In the present study, the authors focused on the rationale for and the accuracy (sensitivity, specificity, and positive and negative predictive values) of IONM in ST surgery in light of more recent evidence in the literature, with specific emphasis on the role of IONM in reducing the incidence of postoperative neurological deficits. This review confirms the role of IONM as a useful tool in the workup for ST surgery. Individual monitoring and mapping techniques are clearly not sufficient to account for the complex function of the SC and NRs. Conversely, multimodal IONM is highly sensitive and specific for anticipating neurological injury during ST surgery and represents an important tool for preserving neuronal structures and achieving an optimal postoperative functional outcome.


Combination of gastric atrophy, reflux symptoms and histological subtype indicates two distinct aetiologies of gastric cardia cancer.
Atrophic gastritis is a risk factor for non-cardia gastric cancer, and gastro-oesophageal reflux disease (GORD) for oesophageal adenocarcinoma. The role of atrophic gastritis and GORD in the aetiology of adenocarcinoma of the cardia remains unclear. We have investigated the association between adenocarcinoma of the different regions of the upper gastrointestinal tract and atrophic gastritis and GORD symptoms. 138 patients with upper GI adenocarcinoma and age- and sex-matched controls were studied. Serum pepsinogen I/II was used as a marker of atrophic gastritis and categorised to five quintiles. History of GORD symptoms, smoking and H pylori infection were incorporated in logistic regression analysis. Lauren classification of gastric cancer was used to subtype gastric and oesophageal adenocarcinoma. Non-cardia cancer was associated with atrophic gastritis but not with GORD symptoms; 55% of these cancers were intestinal subtype. Oesophageal adenocarcinoma was associated with GORD symptoms, but not with atrophic gastritis; 84% were intestinal subtype. Cardia cancer was positively associated with both severe gastric atrophy [OR, 95% CI: 3.92 (1.77 to 8.67)] and with frequent GORD symptoms [OR, 95% CI: 10.08 (2.29 to 44.36)] although the latter was only apparent in the non-atrophic subgroup and in the intestinal subtype. The association of cardia cancer with atrophy was stronger for the diffuse versus intestinal subtype and this was the converse of the association observed with non-cardia cancer. These findings indicate two distinct aetiologies of cardia cancer, one arising from severe atrophic gastritis and being of intestinal or diffuse subtype similar to non-cardia cancer, and one related to GORD and intestinal in subtype, similar to oesophageal adenocarcinoma. Gastric atrophy, GORD symptoms and histological subtype may distinguish between gastric versus oesophageal origin of cardia cancer.


Nonsuicidal Self-Injury-Related Differences in the Experience of Negative and Positive Emotion.
Emotional experience is argued to contribute to the initiation and maintenance of nonsuicidal self-injury (NSSI). We investigated whether individuals with/without a history of NSSI differed in their dispositional experience of negative and positive emotion, as well as their state responses to negatively and positively valenced movie clips. Undergraduates (n = 214, Mage = 21.33, 73.8% female, 35.5% reporting NSSI) completed measures of NSSI and dispositional emotional experience. Participants also viewed a sad and amusing movie clip and provided sadness/amusement ratings at seven time-points. Relative to participants with no history of self-injury, participants reporting NSSI indicated more reactivity, intensity, and perseveration of dispositional negative emotion; however, differences were negated after adjusting for mental illness. Unexpectedly, individuals with a history of NSSI responded less intensely to the sad clip, although they demonstrated perseveration of sadness over time. Participants reporting NSSI also indicated less reactivity, intensity, and perseveration of dispositional positive emotion and, in response to the amusing film, reported less amusement at all time-points. Considering different dimensions of negative and positive emotion may enhance understanding of NSSI. Future research should disentangle which dimensions of emotional experience are unique to NSSI and which are shared with mental illness more generally.


Moderating Effect of Cortical Thickness on BOLD Signal Variability Age-Related Changes.
The time course of neuroanatomical structural and functional measures across the lifespan is commonly reported in association with aging. Blood oxygen-level dependent signal variability, estimated using the standard deviation of the signal, or "BOLDSD," is an emerging metric of variability in neural processing, and has been shown to be positively correlated with cognitive flexibility. Generally, BOLDSD is reported to decrease with aging, and is thought to reflect age-related cognitive decline. Additionally, it is well established that normative aging is associated with structural changes in brain regions, and that these predict functional decline in various cognitive domains. Nevertheless, the interaction between alterations in cortical morphology and BOLDSD changes has not been modeled quantitatively. The objective of the current study was to investigate the influence of cortical morphology metrics [i.e., cortical thickness (CT), gray matter (GM) volume, and cortical area (CA)] on age-related BOLDSD changes by treating these cortical morphology metrics as possible physiological confounds using linear mixed models. We studied these metrics in 28 healthy older subjects scanned twice at approximately 2.5 years interval. Results show that BOLDSD is confounded by cortical morphology metrics. Respectively, changes in CT but not GM volume nor CA, show a significant interaction with BOLDSD alterations. Our study highlights that CT changes should be considered when evaluating BOLDSD alternations in the lifespan.


Investigation of NAD(P)H:quinone oxidoreductase 1 (NQO1) C609T polymorphism in prostate cancer.
NAD(P)H:quinone oxidoreductase 1 (NQO1) is a part of the antioxidant defense system. NQO1 protects cells from oxidative stress by maintaining antioxidant forms of ubiquinone and vitamin E and this enzymatic activity can be sufficient to protect against carcinogenesis. Oxidative stress may contribute to carcinogenesis as an important risk factor. This study aimed to investigate the relationship between prostate cancer and NQO1 C609T polymorphism in a Turkish population. Forty-five patients with prostate cancer and fifty healthy controls were included in this study. Gene polymorphism was determined by a restricted fragment length polymorphism-polymerase chain reaction (PCR-RFLP) method. The NQO1 TT genotype was demonstrated to be increased in patients, there were no significant differences between distributions of NQO1 C609T genotypes in the study groups. Serum PSA and alkaline phosphatase levels were elevated in patients carrying T alleles (TT > CT > CC). There were no correlations between tumor size, node classification, metastasis or stage and NQO1 genotypes. The results of our study of Turkish prostate cancer patients suggests that mutation of the NQO1 gene may effect the serum PSA and alkaline phosphatase levels. However there were no differences between the NQO1 genotypes in the study groups.


Genetic and clinical profile of Indian patients of idiopathic restrictive cardiomyopathy with and without hypertrophy.
Both idiopathic restrictive cardiomyopathy (IRCM) and hypertrophic cardiomyopathy (HCM) are part of the same disease spectrum and are due to sarcomeric gene mutations. A patient with restrictive physiology without left ventricular hypertrophy (LVH) would be diagnosed as IRCM, while one with LVH would be diagnosed as HCM with restrictive physiology. We studied a group of patients with restrictive physiology for mutations in beta-myosin heavy chain (MYH7) and troponin I (TNNI3) gene. Consecutive probands in the HCM and IRCM cohort over a 4-year period were considered for this study. These included 10 IRCM and 102 HCM patients. All were Asian Indians. Among the 17 patients who had restrictive physiology 10 were IRCM patients and seven were HCM patients. Of the HCM patients, seven (6.9%) had restrictive physiology. Mean age of these 17 patients was 40.1 +/- 19.2 years (range: 15-67 ), six (35.3%) were males. Maximal left ventricular wall thickness of the seven HCM probands was 20.7 +/- 5.2 mm (range: 16-31), while it was normal in the IRCM probands. Ten probands (58.8%) were in NYHA class III or IV. Seven patients (41.2%) had atrial fibrillation. All the probands were screened for mutations in selected exons of MYH7 and TNNI3 genes. One IRCM patient was found to have p.Arg721Lys mutation in the MYH7 gene. She died due to progressive congestive cardiac failure at the age of 47 years. One HCM proband with a maximal left ventricular wall thickness of 17 mm had p.Arg192His mutation in the TNNI3 gene. She had features consistent with restrictive physiology. Her father and sister had died of restrictive cardiomyopathy. IRCM and HCM with restrictive physiology, both are part of the clinical expression of MYH7 and TNNI3 mutations and lead to worse clinical onset and progression of the disease.


Awake intranasal insulin delivery modifies protein complexes and alters memory, anxiety, and olfactory behaviors.
The role of insulin pathways in olfaction is of significant interest with the widespread pathology of diabetes mellitus and its associated metabolic and neuronal comorbidities. The insulin receptor (IR) kinase is expressed at high levels in the olfactory bulb, in which it suppresses a dominant Shaker ion channel (Kv1.3) via tyrosine phosphorylation of critical N- and C-terminal residues. We optimized a 7 d intranasal insulin delivery (IND) in awake mice to ascertain the biochemical and behavioral effects of insulin to this brain region, given that nasal sprays for insulin have been marketed notwithstanding our knowledge of the role of Kv1.3 in olfaction, metabolism, and axon targeting. IND evoked robust phosphorylation of Kv1.3, as well as increased channel protein-protein interactions with IR and postsynaptic density 95. IND-treated mice had an increased short- and long-term object memory recognition, increased anxiolytic behavior, and an increased odor discrimination using an odor habituation protocol but only moderate change in odor threshold using a two-choice paradigm. Unlike Kv1.3 gene-targeted deletion that alters metabolism, adiposity, and axonal targeting to defined olfactory glomeruli, suppression of Kv1.3 via IND had no effect on body weight nor the size and number of M72 glomeruli or the route of its sensory axon projections. There was no evidence of altered expression of sensory neurons in the epithelium. In mice made prediabetic via diet-induced obesity, IND was no longer effective in increasing long-term object memory recognition nor increasing anxiolytic behavior, suggesting state dependency or a degree of insulin resistance related to these behaviors.


Treatment of metastatic cancer with tetrathiomolybdate, an anticopper, antiangiogenic agent: Phase I study.
Preclinical and in vitro studies have determined that copper is an important cofactor for angiogenesis. Tetrathiomolybdate (TM) was developed as an effective anticopper therapy for the initial treatment of Wilson's disease, an autosomal recessive disorder that leads to abnormal copper accumulation. Given the potency and uniqueness of the anticopper action of TM and its lack of toxicity, we hypothesized that TM would be a suitable agent to achieve and maintain mild copper deficiency to impair neovascularization in metastatic solid tumors. Following preclinical work that showed efficacy for this anticopper approach in mouse tumor models, we carried out a Phase I clinical trial in 18 patients with metastatic cancer who were enrolled at three dose levels of oral TM (90, 105, and 120 mg/day) administered in six divided doses with and in-between meals. Serum ceruloplasmin (Cp) was used as a surrogate marker for total body copper. Because anemia is the first clinical sign of copper deficiency, the goal of the study was to reduce Cp to 20% of baseline value without reducing hematocrit below 80% of baseline. Cp is a reliable and sensitive measure of copper status, and TM was nontoxic when Cp was reduced to 15-20% of baseline. The level III dose of TM (120 mg/ day) was effective in reaching the target Cp without added toxicity. TM-induced mild copper deficiency achieved stable disease in five of six patients who were copper deficient at the target range for at least 90 days.


The psychosocial health of children born after medically assisted reproduction: Evidence from the UK Millennium Cohort Study.
The increasing number of children conceived through medically assisted reproduction (MAR, including IVF/ICSI, intrauterine insemination and ovulation induction) has led to concerns about the potential negative effects of fertility treatments on children's psychosocial health. Some studies suggest that MAR children might be at higher risk of developing psychosocial problems when they enter adolescence. However, very few studies have examined the development of MAR children after childhood. Moreover, even though parental socio-economic characteristics are known to be highly correlated with children's psychosocial development, most existing studies on the outcomes of MAR children did not take into account the selective characteristics of the couples who accessed fertility treatments. Using data from waves 1-6 of the UK Millennium Cohort Study, we compare the psychosocial health, as measured by the Strengths and Difficulties Questionnaire, of MAR children to that of naturally-conceived (NC) children, up to and including the age of 14. We control for a wide range of time-constant child and parental characteristics that might confound the association between MAR and the psychosocial health of children. Results from multilevel random intercept models that do not account for parental characteristics show that MAR children have a lower incidence of psychosocial problems than NC children. In models that control for parental characteristics, MAR children are found to have a higher incidence of psychosocial problems than NC children at age three, which suggests that high parental resource levels both explain the advantage of MAR children in unadjusted models, and mask the potentially adverse effects of MAR at young ages. However, in the fully adjusted models in which MAR children have more psychosocial problems at young age, the differences with respect to NC children decrease with age and become statistically and substantively negligible by end of follow-up at age 14. This result suggests that the use of MAR does not increase children's risk of having psychosocial problems at the onset of adolescence.


Pretargeting in tumored mice with radiolabeled morpholino oligomer showing low kidney uptake.
We have recently shown that accumulation in mouse kidneys of technetium-99m labeled phosphorodiamidate morpholinos (MORFs) increases with the number of cytosines in the base sequence. To improve tumor/kidney ratios in tumored mice, pretargeting studies were performed with a cytosine-free MORF. An 18-mer MORF (5'-TCTTCTACTTCACAACTA) was conjugated to the anti-CEA antibody MN14 (Immunomedics) and administered to nude mice bearing LS174T tumors. Thereafter, the (99m)Tc-labeled cytosine-free cMORF (5'-TAGTTGTGAAGTAGAAGA-amide-MAG(3)) was administered. For comparison, the identical study was repeated but with our original pair of 18-mer MORFs (5'-GGGTGTACGTCACAACTA-conjugated MN14 and (99m)Tc-labeled 5'-TAGTTGTGACGTACACCC-amide-MAG(3)). Surface plasmon resonance was used to show that the hybridization affinities of the original and the modified pair of MORFs were essentially equal. Hybridization of the cytosine-free cMORF-(99m)Tc to MN14-MORF was demonstrated in vitro by size-exclusion high-performance liquid chromatography. At 3 h, kidney levels in normal mice were 2.0%ID/organ for the modified cMORF vs. 4.1%ID/organ for the original cMORF sequence, while at 24 h, these values were 0.9% vs 1.8%ID/organ. Pretargeting studies in tumored mice receiving 25 microg of conjugated antibody, 0.5 microg of labeled cMORF 48 h later, followed by imaging and sacrifice at 3 h showed that kidney levels were reduced using the cytosine-free cMORF. Moreover, tumor accumulation was about 3.6%ID/g and was independent of sequence. The whole-body images clearly reflected the improved tumor to kidney ratios. By choosing a cytosine-free base sequence for pretargeting studies, kidney accumulation of cMORF-(99m)Tc was reduced without adversely influencing tumor accumulation. The lowering of kidney radioactivity levels in this way may be important to reduce toxicity to this organ in connection with pretargeting radiotherapy studies.


Deciphering the role of Shh signaling in axial defects produced by ethanol exposure.
The phenotype of embryos exposed to ethanol is complex and likely due to multiple alterations in developmental pathways. We have previously demonstrated that Sonic hedgehog signaling (Shh-s) was reduced in both chicken and zebrafish embryos when exposed to ethanol. There are many tissues affected by embryonic ethanol exposure, and in this article we explore the development of axial tissues, using zebrafish embryos. We then compare these effects to the phenotypes produced by exposure to two drugs that also inhibit Shh-s: cyclopamine and forskolin. We found alterations in the development of the notochord and somites produced by all three compounds, although only ethanol produced developmental delay of epiboly. Upon observation of early developing embryos, muscle pioneer cells were completely lost in cyclopamine-treated embryos, and reduced, but less so, in embryos treated with forskolin and ethanol. Ethanol treatment produced a dose-dependent reduction in total body length that may be linked to epiboly delay seen earlier during development. Despite the differences between cyclopamine and forskolin, we found that shh mRNA injection rescued the short body length, the alteration in somite shape, and the cyclopia produced by ethanol exposure. Taken together, each teratogen produced a unique set of phenotypic changes in the body axis, suggesting that each compound affects Shh-s and also produces a distinctive set of molecular alterations. However, addition of exogenous Shh to ethanol treated zebrafish prevented many of the gross physical phenotypes, suggesting that the suppression of Shh-s is one of the major effects of ethanol exposure.


The Role of Social Media and Internet Search Engines in Information Provision and Dissemination to Patients with Kidney Stone Disease: A Systematic Review from European Association of Urologists Young Academic Urologists.
Kidney stone disease (KSD) affects millions of people worldwide and has an increasing incidence. Social media (SoMe) and search engines are both gaining in usage, while also being used by patients to research their conditions and aid in managing them. With this in mind, many authors have expressed the belief that SoMe and search engines can be used by patients and healthcare professionals to improve treatment compliance and help counseling and management of conditions such as KSD. We wanted to determine whether SoMe and search engines play a role in the management and/or prevention of KSD. The databases MEDLINE, Embase, CINAHL, Scopus, and Cochrane Library were used to search for relevant English language literature from inception to December 2017. Results were screened by title, abstract, and then full text, according to the inclusion and exclusion criteria. The data were then analyzed independently by the authors not involved in the original study. After initial identification of 2137 records and screening of 42 articles, 10 studies met the inclusion and exclusion criteria. The articles included focused on a variety of SoMe forms, including two articles each on twitter, YouTube, smartphone apps, and Google search engine and one article on Google insights and Google analytics. Regarding patient centered advice, while two articles covered advice on dietary, fluid intake, and management options, two additional articles each covered advice on fluid advice and management options only, while no such advice was given by three of the SoMe published articles. SoMe and search engines provide valuable information to patients with KSD. However, while the information provided regarding dietary aspects and fluid management was good, it was not comprehensive enough to include advice on other aspects of KSD prevention.


Association of common LH variant with hyperfunctional promoter in a Japanese infertile woman.
A common LH variant (V-LH) with Trp8Arg and Ile15Thr is often associated with ovarian dysfunction primarily in the Japanese population, and the LHB gene encoding V-LH is linked with a hyperfunctional promoter that could partly compensate for the somewhat weak biological effect of the V-LH in the Finnish and other several populations. We analyzed the promoter region in a Japanese infertile woman homozygous for the V-LH, to examine whether the hyperfunctional promoter is present or absent in the Japanese V-LH carriers with ovarian dysfunction. Direct sequencing was performed for a 661 bp promoter region from -8 to -668 bp of LHB, revealing homozygosity for eight nucleotide substitutions (-238A>G, -276G>A, -489C>A, -490T>A, -504T>A, -506T>C, -525T>G, and -552C>T) that are identical to those found in the hyperfunctional promoter. The results suggest that ovarian dysfunction frequently observed in the Japanese V-LH carriers would be due to some population-specific genetic and/or environmental factor(s) rather than to the lack of the hyperfunctional promoter and the resultant low biological effect of the V-LH. In addition, the tight linkage between the two missense substitutions in the coding region and the eight nucleotide substitutions in the promoter region of LHB appears to be common to various ethnic groups.


Genetic diversity of beta-thalassemia mutations in Pakistani population.
beta-thalassemia is one of the most common inherited single gene disorder in Pakistan. It is characterized by reduced or absent beta-globin gene expression resulting in abnormal maturation and survival of red blood cells. Due to high prevalence of this disease in the local population, it has become important for the health care providers to encourage people to utilize laboratory facilities for carrier and prenatal genetic testing. To study the frequency of beta-thalassemia mutations in Pakistani population. A tertiary care teaching hospital. Blood samples of 72 couples and chorionic villus (CV) biopsy specimen collected at the Aga Khan University Hospital, Karachi were tested by Amplified Refractory Mutation Systems (ARMS) for the 12 most common mutations in the beta-globin gene. Out of 72 chorionic villus biopsy specimen analyzed, 17 (23%) had mutations in both alleles of the beta-globin gene. Homozygosity was identified in 6 CV samples, whereas 11 CV specimens were diagnosed as double heterozygous. Almost 60% of the CV biopsies showed mutations in one allele and were diagnosed as carriers. IVSI-5 (G-C) was the most common mutation identified in this study. It was found in 53% of the subjects and was represented equally in all the ethnic groups except Pathans. Several regional and ethnic differences were observed in the distribution of common mutations, for example in Pathan families Fr 8-9 (+G) mutation was most prevalent. In addition, variation in the distribution of mutations was also observed between the Northern and the Southern regions. This study indicates that in Pakistan, the five most common mutations are IVS1-5 (G-C), IVS1-1 (G-T), Fr 41-42 (-TTCT) Fr 8-9 (+G) and deletion 619 bp. An important factor contributing to high incidence of thalassemia is the unawareness among people about the available diagnostic facilities for the prenatal diagnosis in Pakistan. Strict implementation of collective measures including carrier identification, genetic counseling and prenatal diagnosis are required for preventing beta-thalassemia in Pakistan.


Anti-angiogenic therapies in cancer clinical trials.
Strategies involving vasculature have widely been acknowledged to have therapeutic potential in the management of cancer and other diseases. Based on a large body of evidence from preclinical studies and early clinical trials there is considerable optimism that anti-angiogenesis and vascular targeting will be a major clinical therapy. This review considers some 30 anti-angiogenic and vascular targeting agents that are currently in cancer clinical trials and highlights specific problems relating to the assessment of the activity of these agents in patients, trial design, potential toxicities and resistance mechanisms.


The interrelationships between working conditions, job satisfaction, burnout and mental health among hospital physicians in Japan: a path analysis.
A growing number of physicians are leaving their hospitals because of painful working conditions in hospitals throughout Japan. We set out to analyze the interrelationships between working conditions, job satisfaction, burnout and mental health among Japanese physicians. A cross-sectional survey was conducted in 2007 for hospital physicians throughout Japan. A path analysis based on structural equation modeling was utilized for examining the interrelationships between work control, on-call duty volume, job satisfaction (the Japan Hospital Physicians Satisfaction Scale), burnout (the Japanese version of the Maslach Burnout Inventory) and mental health (the General Health Questionnaire). Of 336 physicians who received a solicitation E-mail, 236 participated in our study (response rate, 70%). Sixty physicians (25.4%) were women with a mean age of 41 yr. In the path analysis, burnout and poor mental health were related directly to job dissatisfaction and short sleeping time, while they were related indirectly to poor work control and heavy on-call duty. In the multi-group path analysis of both genders, sleeping time was related to job satisfaction more likely among female physicians but less among male physicians. Healthcare policy makers need to implement immediate, extensive and decisive measures to improve work condition and to reduce overwork among hospital physicians.