Predicting physical activity energy expenditure using accelerometry in adults from sub-Sahara Africa.
Lack of physical activity may be an important etiological factor in the current epidemiological transition characterized by increasing prevalence of obesity and chronic diseases in sub-Sahara Africa. However, there is a dearth of data on objectively measured physical activity energy expenditure (PAEE) in this region. We sought to develop regression equations using body composition and accelerometer counts to predict PAEE. We conducted a cross-sectional study of 33 adult volunteers from an urban (n = 16) and a rural (n = 17) residential site in Cameroon. Energy expenditure was measured by doubly labeled water (DLW) over a period of seven consecutive days. Simultaneously, a hip-mounted Actigraph accelerometer recorded body movement. PAEE prediction equations were derived using accelerometer counts, age, sex, and body composition variables, and cross-validated by the jack-knife method. The Bland and Altman limits of agreement (LOAs) approach was used to assess agreement. Our results show that PAEE (kJ/kg/day) was significantly and positively correlated with activity counts from the accelerometer (r = 0.37, P = 0.03). The derived equations explained 14-40% of the variance in PAEE. Age, sex, and accelerometer counts together explained 34% of the variance in PAEE, with accelerometer counts alone explaining 14%. The LOAs between DLW and the derived equations were wide, with predicted PAEE being up to 60 kJ/kg/day below or above the measured value. In summary, the derived equations performed better than existing published equations in predicting PAEE from accelerometer counts in this population. Accelerometry could be used to predict PAEE in this population and, therefore, has important applications for monitoring population levels of total physical activity patterns.


COVID-19 in a Hispanic Woman.
Since making its debut on the global stage in December 2019, coronavirus disease 2019 (COVID-19) has afflicted nearly 4 million people and caused hundreds of thousands of deaths. Case reports and case series depicting the clinical effects of the causative virus-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-have been published, yet few demonstrate the cytopathologic alterations of this disease. We present a clinical-pathologic correlation report of a previously healthy Hispanic woman with laboratory-confirmed COVID-19 who had typical features of acute respiratory distress syndrome (ARDS) and also showed cardiac abnormalities thought to represent fulminant viral myocarditis. Congruent with the ARDS clinical impression, autopsy findings were remarkable for extensive and markedly severe acute lung injury consistent with viral pneumonia, characterized by diffuse alveolar damage, pulmonary infarction, severe pulmonary edema, desquamation of pneumocytes with intra-alveolar aggregation, and pneumocyte morphologic alterations suggestive of viral cytopathic effect. However, there was incongruence between the clinical impression and the cardiovascular pathology findings in that viral myocarditis was not detected on histopathologic evaluation. This case highlights the importance of pathologic corroboration of the clinical impression and, in addition, illuminates the key role autopsy plays during a pandemic by providing valuable insight into viral pathology in tissues.


Stapling for wound dehiscence after cardiac implantable electronic device implantation.
Wound dehiscence (WD) has been reported as a complication in 0.3% of cardiac implantable electronic device (CIED) procedures. Stapling has not previously been reported as a treatment modality for WD. Presently described is the experience of a single center with WD and its management. A retrospective chart review of all patients who underwent CIED implantation between 2009 and 2016, a total of 759 devices, was performed. There were a total of 11 (1.4%) patients with WD. The majority 9/11 patients were female, 5 of 11 (45.5%) had diabetes, and 2 of the 11 patients were immunocompromised due to recent chemotherapy. WD occurred in 6 patients after generator change, in 2 patients after a biventricular device upgrade, in 1 patient after biventricular implantable cardioverter defibrillator (ICD) implantation, in 1 patient after dual-chamber pacemaker implantation, and in 1 patient after subcutaneous ICD implantation. The median time of WD was 6 weeks post procedure (range: 1-20 weeks). In all of the patients, wound stapling was performed under sterile conditions after administering intravenous narcotic analgesics. Eight patients received intravenous antibiotics and all patients received at least 2 weeks of oral antibiotics. Blood cultures were negative in 8/11 (72.7%) patients. However, the wound cultures in 5 patients were positive. The staples were removed in a median of 16 days (range: 9-36 days). All of these patients were successfully treated with stapling and none of the devices required extraction. Stapling under sterile conditions may be an acceptable treatment strategy to manage WD after device implantation. This can be performed as an outpatient procedure and can help avoid unnecessary device extraction.


Molecular diagnosis of Toxoplasma gondii infection in Libya.
Toxoplasma gondii infections are prevalent in humans and animals throughout Libya. Current diagnosis is based on detection of Toxoplasma-specific IgM and IgG. In this study, we established and optimized a diagnostic PCR assay for molecular diagnosis of T. gondii in Libya. From January to December, 2010, 177 blood and serum samples were collected from suspected patients. This includes: 140 women who have had spontaneous abortions, 26 HIV-positive patients, nine patients with leukemia and lymphoma, and two infants with ocular infection. Samples were screened for anti-Toxoplasma IgG and IgM antibodies before DNA extraction. The surface antigen gene 2 (SAG2) was targeted in a semi-nested PCR to amplify a 999 bp and a 614 bp fragment in the first and the second run respectively. A total of 54/140 (38.5 %) women who have had spontaneous abortions, 23/26 (88 %) HIV patients, 6/9 (66.6 %) of the leukaemia and lymphoma patients, and one child with ocular infection were seropositive for anti-Toxoplasma IgG and/or IgM. Genomic DNA was extracted from 38 selected seropositive samples. The PCR was sensitive enough to detect DNA concentration of 12 ng/μL. PCR analysis was performed for 38 selected seropositive patients (16 women who have had spontaneous abortions, 15 positive HIV patients, six leukaemia patients and one child with ocular infection). Our designed primers were successfully amplified in 22/38 (57.9 %) samples; 5/12 (35.7 %) from serum and 17/26 (65.8 %) from whole blood samples. All PCR positive samples were IgG-positive except two samples which were IgM and IgG & IgM-positive serum samples respectively. The semi-nested PCR confirmed five more samples. These included two leukaemia and two HIV-positive whole blood samples and one serum sample from an aborted woman. The ability of PCR to diagnose active toxoplasmosis is needed in immunocompromised patients and congenital toxoplasmosis cases, especially when serological techniques fail. For the first time in Libya, we established and optimized semi-nested PCR of SAG2 gene. The developed PCR method was able to detect as little as 12 ng/μL of T. gondii DNA and was useful to diagnose the diseases in women who have had spontaneous abortions, HIV-positive patients, patients with leukemia and lymphoma, and infants with ocular infection.


Carotid body adaptation to hypoxia: cellular and molecular mechanisms in vitro.
Chronic hypoxia in vivo promotes long-term changes in the carotid body (CB) response to low PO2. By exposing cultured rat CB chemoreceptors (glomus cells) to 6% O2 for 1-3 weeks, we are investigating the cellular and molecular mechanisms of hypoxic adaptation. Recent studies have uncovered a series of plastic changes in glomus cells including hypertrophy, differential regulation of Na+, Ca2+, and K+ currents, and upregulation of the 'plasticity protein', GAP-43. We have also identified cyclic AMP as a possible intracellular mediator of at least some of these effects of chronic hypoxia. Associated with the changes in ionic currents, glomus cells become electrically more excitable. However, a complete understanding of the physiological response of chronically hypoxic glomus cells to chemostimuli will require more detailed knowledge of the specific alterations in the sensing and signaling pathways, including modifications in neurotransmitter (e.g. catecholamine) functions.


DNA ligand designed to antagonize EBNA1 represses Epstein-Barr virus-induced immortalization.
Epstein-Barr virus (EBV) transforms human B lymphocytes into immortalized cells in vitro and is associated with various malignancies in vivo. EBNA1, which is expressed in the majority of EBV-infected cells, recognizes specific DNA sequences at the cis-acting latent origin of plasmid replication (oriP) element of the EBV genome. EBNA1 plays a critical role in the viral episome maintenance and transactivates viral transforming genes in latently infected cells. Therefore, DNA-targeting agents that can disrupt the EBNA1-oriP interaction will offer novel functional inhibitors of EBNA1. Pyrrole-imidazole polyamides, sequence-specific DNA ligands, can be designed to interfere with the binding of various transcriptional factors. Here, we synthesized pyrrole-imidazole polyamides targeting EBNA1-bound DNA sequences and developed an inhibitor for the EBNA1-oriP interaction. A pyrrole-imidazole polyamide, designated as DSE-3, bound adjacent to the EBNA1 recognition sequences located in the dyad symmetry element of oriP, and selectively inhibited EBNA1-oriP binding both in vitro and in vivo. DSE-3 also inhibited the proliferation of established lymphoblastoid cell lines by eradicating EBV episomes from the cells. In addition, DSE-3 repressed the expression of viral transforming genes after infecting human peripheral blood mononuclear cells with EBV and, as a consequence, inhibited EBV-mediated B-cell immortalization. These results suggest that EBNA1 functions will be an attractive pharmacological target for EBV-associated diseases.


[Evaluation of the immune response after vaccination against distemper at a mink (Mustela vison) farm in Argentina].
Distemper virus causes a disease affecting minks with respiratory, gastrointestinal, neurological and skin symptoms and showing high morbidity and mortality, mainly among puppies. It is controlled through immunization, using vaccines that are supplied for mink use. The aim of this work was to determine the seroneutralization titer against the distemper virus at a mink farm in Argentina. The antibody kinetics obtained after vaccination in 27 adult animals, as well as the duration of colostrum-transferred antibodies in 10 puppies were determined. All vaccinated adult minks showed protective titers up to at least 3 months after vaccination, and 37.5% significantly reduced their antibody levels, 12 months after vaccination. Only 20% of the puppies showed protective levels of colostrum-transferred antibodies at the age of 7 weeks, while non-detectable levels of antibodies were found when puppies reached 11 weeks old. Vaccination performed in these puppies at the age of 13 weeks, elicited protective seroneutralization titers. These results show that vaccination induces a satisfactory humoral immune response in our environment, and support the convenience of vaccinating dams annually before the beginning of the breeding season. The vaccination plan in puppies is also discussed.


Acute Liver Failure and Myelosuppression due to Parvovirus B19 Infection: A Case Report.
Parvovirus B19 has been associated with different diseases, such as erythema infectiosum, arthropathy and transient aplastic crisis. However, parvovirus B19 infection presenting as hepatic dysfunction and myelosuppression is rarely reported in adult patients. Herein, we report an adult case of acute parvovirus B19 infection presented with acute liver failure and myelosuppression. After being treated with gamma globulin, the patient's liver function and bone marrow test improved. We conclude that the parvovirus B19 infection should be considered as a possible cause of acute liver injury and bone marrow suppression. The antibody for B19 virus should be routinely tested in patients with liver dysfunction and/or myelosuppression of unclear etiology.


A synthetic antibody fragment targeting nicastrin affects assembly and trafficking of γ-secretase.
The γ-secretase complex, composed of presenilin, nicastrin (NCT), anterior pharynx-defective 1 (APH-1), and presenilin enhancer 2 (PEN-2), is assembled in a highly regulated manner and catalyzes the intramembranous proteolysis of many type I membrane proteins, including Notch and amyloid precursor protein. The Notch family of receptors plays important roles in cell fate specification during development and in adult tissues, and aberrant hyperactive Notch signaling causes some forms of cancer. γ-Secretase-mediated processing of Notch at the cell surface results in the generation of the Notch intracellular domain, which associates with several transcriptional coactivators involved in nuclear signaling events. On the other hand, γ-secretase-mediated processing of amyloid precursor protein leads to the production of amyloid β (Aβ) peptides that play an important role in the pathogenesis of Alzheimer disease. We used a phage display approach to identify synthetic antibodies that specifically target NCT and expressed them in the single-chain variable fragment (scFv) format in mammalian cells. We show that expression of a NCT-specific scFv clone, G9, in HEK293 cells decreased the production of the Notch intracellular domain but not the production of amyloid β peptides that occurs in endosomal and recycling compartments. Biochemical studies revealed that scFvG9 impairs the maturation of NCT by associating with immature forms of NCT and, consequently, prevents its association with the other components of the γ-secretase complex, leading to degradation of these molecules. The reduced cell surface levels of mature γ-secretase complexes, in turn, compromise the intramembranous processing of Notch.


A survey of consultants and specialists in restorative dentistry on their use of health-related quality of life questionnaires.
A national survey of 178 consultants and specialists in restorative dentistry was undertaken to assess their use of health-related quality of life questionnaires in patients having oral rehabilitation following treatment for oral cancer. The response rate was 74% (132). 42% treated patients following oral cancer, 25% forming part of the head and neck multidisciplinary team. Only 19% had ever used questionnaires. Main barriers to their use were lack of experience with evaluation (27%), lack of staff to administer questionnaires (21%) and insufficient time (18%). Few clinicians thought that questionnaires were not clinically relevant or that no suitable instrument existed. In order to facilitate the use of an oral rehabilitation-specific questionnaire in clinical practice there needs to be greater awareness of the potential benefits, training in the process and the means of reducing the administrative burden.


Role-Playing and Real-Time Strategy Games Associated with Greater Probability of Internet Gaming Disorder.
Research indicates that a small subset of those who routinely play video games show signs of pathological habits, with side effects ranging from mild (e.g., being late) to quite severe (e.g., losing a job). However, it is still not clear whether individual types, or genres, of games are most strongly associated with Internet gaming disorder (IGD). A sample of 4,744 University of Wisconsin-Madison undergraduates (Mage=18.9 years; SD=1.9 years; 60.5% female) completed questionnaires on general video game playing habits and on symptoms of IGD. Consistent with previous reports: 5.9-10.8% (depending on classification criteria) of individuals who played video games show signs of pathological play. Furthermore, real-time strategy and role-playing video games were more strongly associated with pathological play, compared with action and other games (e.g., phone games). The current investigation adds support to the idea that not all video games are equal. Instead, certain genres of video games, specifically real-time strategy and role-playing/fantasy games, are disproportionately associated with IGD symptoms.


Dissecting the order of bacteriophage T4 DNA polymerase holoenzyme assembly.
Most biological organisms rely upon a DNA polymerase holoenzyme for processive DNA replication. The bacteriophage T4 DNA polymerase holoenzyme is composed of the polymerase enzyme and a clamp protein (the 45 protein), which functions as a processivity factor by strengthening the interaction between DNA and the holoenzyme. The 45 protein must be loaded onto DNA by a clamp loader ATPase complex (the 44/62 complex). In this paper, the order of events leading to holoenzyme formation is investigated using a combination of rapid-quench and stopped-flow fluorescence spectroscopy kinetic methods. A rapid-quench strand displacement assay in which the order of holoenzyme component addition is varied provided data indicating that the rate-limiting step in holoenzyme assembly is associated with the clamp loading process. Pre-steady-state analysis of the clamp loader ATPase activity demonstrated that the four bound ATP molecules are hydrolyzed stepwise during the clamp loading process in groups of two. Clamp loading was examined with stopped-flow fluorescence spectroscopy from the perspective of the clamp itself, using a site-specific, fluorescently labeled 45 protein. A mechanism for T4 DNA polymerase holoenzyme assembly is proposed in which the 45 protein interacts with the 44/62 complex leading to the hydrolysis of 2 equiv of ATP, and upon contacting DNA, the remaining two ATP molecules bound to the 44/62 complex are hydrolyzed. Once all four ATP molecules are hydrolyzed, the 45 protein is poised on DNA for association with the polymerase to form the holoenzyme.


Jabba: hybrid error correction for long sequencing reads.
Third generation sequencing platforms produce longer reads with higher error rates than second generation technologies. While the improved read length can provide useful information for downstream analysis, underlying algorithms are challenged by the high error rate. Error correction methods in which accurate short reads are used to correct noisy long reads appear to be attractive to generate high-quality long reads. Methods that align short reads to long reads do not optimally use the information contained in the second generation data, and suffer from large runtimes. Recently, a new hybrid error correcting method has been proposed, where the second generation data is first assembled into a de Bruijn graph, on which the long reads are then aligned. In this context we present Jabba, a hybrid method to correct long third generation reads by mapping them on a corrected de Bruijn graph that was constructed from second generation data. Unique to our method is the use of a pseudo alignment approach with a seed-and-extend methodology, using maximal exact matches (MEMs) as seeds. In addition to benchmark results, certain theoretical results concerning the possibilities and limitations of the use of MEMs in the context of third generation reads are presented. Jabba produces highly reliable corrected reads: almost all corrected reads align to the reference, and these alignments have a very high identity. Many of the aligned reads are error-free. Additionally, Jabba corrects reads using a very low amount of CPU time. From this we conclude that pseudo alignment with MEMs is a fast and reliable method to map long highly erroneous sequences on a de Bruijn graph.


Panning of multiple subsets of leukocytes on antibody-decorated poly(ethylene) glycol-coated glass slides.
The antibody (Ab) array format provides a unique opportunity to pan and characterize multiple leukocyte subsets in parallel. However, the questions of reproducibility and robustness of leukocyte panning on Ab arrays need to be answered for this technology to become an immunophenotyping tool. The present study sought to address several of these questions, including: (1) purity of leukocyte subsets captured on Ab regions, (2) dynamics of leukocyte binding, (3) elimination of non-specific cell adhesion, and (4) standardization of cell washing conditions. Abs for CD4 T-cells, CD8 T-cells, CD36 monocytes, and CD16b neutrophils were dispensed onto standard glass slides containing a thin film of poly(ethylene glycol) (PEG) hydrogel. PEG gel coating was highly effective in eliminated non-specific cell adhesion on the surface. Incubation of the Ab arrays with red blood cell (RBC) depleted whole blood resulting in antigen-specific panning of leukocyte subsets on the respective Ab domains. A flow through chamber was employed to determine optimal shear stress conditions for removal of non-specifically attached cells. The purity of the four subsets remaining on the surface after washing was determined by Wright staining and immunofluorescence, and was found to be as follows: CD4 T-cells (99.2+/-0.3%), CD8 T-cells (98.7+/-0.3%), CD36 monocytes (97.2+/-0.9%), and CD16b neutrophils (99.1+/-0.6%). In conclusion, the methods described in this study allow to separate whole blood into pure leukocyte subsets with minimal sample preparation and handling. These approaches will be valuable in the future development of Ab arrays as tools for quantitative immunophenotyping of leukocytes.


Comparative evaluation of risk management frameworks for U.S. source waters.
The U.S. Safe Drinking Water Act required states to develop source water assessment programs identifying existing and potential contamination sources; however, comprehensive risk prioritization and management approaches for surface water supplies have seen limited application. This participatory study assessed which permutation(s) of risk management frameworks and tools might benefit U.S. utilities by combining a literature review with external utility interviews. Qualitative data provided a basis for categorical assignments of goodness of fit with each of 24 framework evaluation criteria across five categories. Weighted integration using stakeholder input provided a relative ranking of applicability, later validated at a decision-making workshop. Hybridization of the American National Standards Institute/American Water Works Association (ANSI/AWWA G300) source water protection standard and World Health Organization Water Safety Plan guidance was recommended to develop a comprehensive risk management approach for U.S. source waters. Cost-benefit components of other guidance materials were recommended to incorporate financial considerations into risk ranking and mitigation decisions.


Estimating the number of UK stroke patients eligible for endovascular thrombectomy.
Endovascular thrombectomy is a highly effective treatment for acute ischemic stroke due to large arterial occlusion. Routine provision will require major changes in service configuration and workforce. An important first step is to quantify the population of stroke patients that could benefit. We estimated the annual UK population suitable for endovascular thrombectomy using standard or advanced imaging for patient selection. Evidence from randomised control trials and national registries was combined to estimate UK stroke incidence and define a decision-tree describing the endovascular thrombectomy eligible population. Between 9620 and 10,920 UK stroke patients (approximately 10% of stroke admissions) would be eligible for endovascular thrombectomy annually. The majority (9140-9620) would present within 4 h of onset and be suitable for intravenous thrombolysis. Advanced imaging would exclude 500 patients presenting within 4 h, but identify an additional 1310 patients as eligible who present later. Information from randomised control trials and large registry data provided the evidence criterion for 9 of the 12 decision points. The best available evidence was used for two decision points with sensitivity analyses to determine how key branches of the tree affected estimates. Using the mid-point estimate for eligibility (9.6% of admissions) and assuming national endovascular thrombectomy coverage, 4280 patients would have reduced disability. A model combining published trials and register data suggests approximately 10% of all stroke admissions in the UK are eligible for endovascular thrombectomy. The use of advanced imaging based on current published evidence did not have a major impact on overall numbers but could alter eligibility status for 16% of cases.


Case studies in terminal weaning from mechanical ventilation.
Terminal weaning, withdrawal of mechanical ventilation when the patient is not expected to survive the process, must not be burdensome to the patient or significant others. The healthcare team must individualize the weaning process, considering the physiologic comfort of the patient and the psychoemotional comfort of both the patient and family. The following case studies illustrate variations in a method for terminal weaning that are patient-specific and are based on the experience of a supportive care team.


Human alveolar macrophage growth factor for fibroblasts. Regulation and partial characterization.
The number of fibroblasts composing the alveolar structures in controlled within narrow limits by a strictly modulated rate of fibroblast replication. One possible source of growth-modulating signals for alveolar fibroblasts is the alveolar macrophage, a member of the mononuclear phagocyte family of cells, which collectively are known to be important sources of growth factors for a variety of target cells. To evaluate the role of alveolar macrophages in the control of alveolar fibroblast replication, macrophages from normal individuals obtained by bronchoalveolar lavage were maintained in suspension culture with and without added stimuli, and supernates were evaluated for fibroblast growth-promoting effect. Supernates from unstimulated macrophages contained no growth factor activity. In marked contrast, supernates from macrophages stimulated with particulates and immune complexes contained a growth factor that caused a significant increase in fibroblast replication rate. Maximum growth factor activity was observed 3-4 h after macrophage stimulation, at a concentration of 1-2 x 10(6) macrophages/ml. The alveolar macrophagederived growth factor eluted from DEAE-cellulose at 0.27 M NaCl at neutral pH had an apparent molecular weight of 18,000, and appeared to be distinct from other characterized growth factors. The alveolar macrophage-derived growth factor stimulated lung fibroblast DNA synthesis within 12 h, with cell division apparent within 48 h. In serum-free culture, the alveolar macrophage-derived growth factor by itself did not promote fibroblast replication, but rather acted as a progression factor causing a synergistic increase in fibroblast replication rate in the presence of competence factors such as fibroblast growth factor or platelet-derived growth factor. These studies suggest that when stimulated, human alveolar macrophages may modulate, in part, the replication rate of alveolar fibroblasts by releasing a growth factor within the alveolar microenvironment.


Tuberculosis mortality and the male survival deficit in rural South Africa: An observational community cohort study.
Women live on average five years longer than men, and the sex difference in longevity is typically lower in populations with high mortality. South Africa-a high mortality population with a large sex disparity-is an exception, but the causes of death that contribute to this difference are not well understood. Using data from a demographic surveillance system in rural KwaZulu-Natal (2000-2014), we estimate differences between male and female adult life expectancy by HIV status. The contribution of causes of death to these life expectancy differences are computed with demographic decomposition techniques. Cause of death information comes from verbal autopsy interviews that are interpreted with the InSilicoVA tool. Adult women lived an average of 10.4 years (95% confidence Interval 9.0-11.6) longer than men. Sex differences in adult life expectancy were even larger when disaggregated by HIV status: 13.1 (95% confidence interval 10.7-15.3) and 11.2 (95% confidence interval 7.5-14.8) years among known HIV negatives and positives, respectively. Elevated male mortality from pulmonary tuberculosis (TB) and external injuries were responsible for 43% and 31% of the sex difference in life expectancy among the HIV negative population, and 81% and 16% of the difference among people living with HIV. The sex differences in adult life expectancy in rural KwaZulu-Natal are exceptionally large, atypical for an African population, and largely driven by high male mortality from pulmonary TB and injuries. This is the case for both HIV positive and HIV negative men and women, signalling a need to improve the engagement of men with health services, irrespective of their HIV status.


Heritable influences on behavioural problems from early childhood to mid-adolescence: evidence for genetic stability and innovation.
Although behavioural problems (e.g., anxiety, conduct, hyperactivity, peer problems) are known to be heritable both in early childhood and in adolescence, limited work has examined prediction across these ages, and none using a genetically informative sample. We examined, first, whether parental ratings of behavioural problems (indexed by the Strengths and Difficulties questionnaire) at ages 4, 7, 9, 12, and 16 years were stable across these ages. Second, we examined the extent to which stability reflected genetic or environmental effects through multivariate quantitative genetic analysis on data from a large (n > 3000) population (UK) sample of monozygotic and dizygotic twins. Behavioural problems in early childhood (age 4 years) showed significant associations with the corresponding behavioural problem at all subsequent ages. Moreover, stable genetic influences were observed across ages, indicating that biological bases underlying behavioural problems in adolescence are underpinned by genetic influences expressed as early as age 4 years. However, genetic and environmental innovations were also observed at each age. These observations indicate that genetic factors are important for understanding stable individual differences in behavioural problems across childhood and adolescence, although novel genetic influences also facilitate change in such behaviours.


Recommendations for the use of structural magnetic resonance imaging in the care of patients with epilepsy: A consensus report from the International League Against Epilepsy Neuroimaging Task Force.
Structural magnetic resonance imaging (MRI) is of fundamental importance to the diagnosis and treatment of epilepsy, particularly when surgery is being considered. Despite previous recommendations and guidelines, practices for the use of MRI are variable worldwide and may not harness the full potential of recent technological advances for the benefit of people with epilepsy. The International League Against Epilepsy Diagnostic Methods Commission has thus charged the 2013-2017 Neuroimaging Task Force to develop a set of recommendations addressing the following questions: (1) Who should have an MRI? (2) What are the minimum requirements for an MRI epilepsy protocol? (3) How should magnetic resonance (MR) images be evaluated? (4) How to optimize lesion detection? These recommendations target clinicians in established epilepsy centers and neurologists in general/district hospitals. They endorse routine structural imaging in new onset generalized and focal epilepsy alike and describe the range of situations when detailed assessment is indicated. The Neuroimaging Task Force identified a set of sequences, with three-dimensional acquisitions at its core, the harmonized neuroimaging of epilepsy structural sequences-HARNESS-MRI protocol. As these sequences are available on most MR scanners, the HARNESS-MRI protocol is generalizable, regardless of the clinical setting and country. The Neuroimaging Task Force also endorses the use of computer-aided image postprocessing methods to provide an objective account of an individual's brain anatomy and pathology. By discussing the breadth and depth of scope of MRI, this report emphasizes the unique role of this noninvasive investigation in the care of people with epilepsy.


Influence of surface area on charge transport and recombination in dye-sensitized TiO2 solar cells.
The dependence of the electron transport and recombination dynamics on the internal surface area of mesoporous nanocrystalline TiO2 films in dye-sensitized solar cells was investigated. The internal surface area was varied by altering the average particle size in the films. The scaling of the photoelectron density and the electron diffusion coefficient at short circuit with internal surface area confirms the results of a recent study (Kopidakis, N.; Neale, N. R.; Zhu, K.; van de Lagemaat, J.; Frank, A. J. Appl. Phys. Lett. 2005, 87, 202106) that transport-limiting traps are located predominately on the surfaces of the particles. The recombination current density was found to increase superlinearly (with an exponent of 1.40 +/- 0.12) with the internal surface area. This result is at odds with the expected linear dependence of the recombination current density on the surface area when only the film thickness is increased. The observed scaling of the recombination current density with surface area is consistent with recombination being transport-limited. Evidence is also presented confirming that photoinjected electrons recombine with redox species in the electrolyte via surface states rather than from the TiO2 conduction band.


Role of intracardiac ultrasound in interventional electrophysiology.
Interventional procedures in the electrophysiology and catheterization laboratory are rapidly advancing. Critical to the advancement of these procedures is accurate identification of critical anatomic landmarks and catheter position. Fluoroscopy remains the mainstay for general identification of anatomic landmarks but is inadequate for the precise imaging needed for complex procedures. Precise imaging of anatomic landmarks and catheter position is now possible during the procedure with the use of intracardiac echocardiography (ICE). This paper reviews the rapid development and utilization of ICE in interventional electrophysiology. Several recent studies show ICE as a major contribution to providing a safer, more reliable, and more cost-effective means of accomplishing the tasks performed by existing techniques. In the electrophysiology laboratory, the dependence on this new technology has been due to the rapid development of catheter-based radiofrequency ablation of the pulmonary veins for treatment of atrial fibrillation. Since the initial use of ICE in facilitating ablation of atrial fibrillation, other uses for ICE are continuously being identified. A comprehensive look is provided at the history and development of this new technology along with the most recent applications of ICE in interventional electrophysiology.


Annual meeting of the Tuberculosis Surveillance and Research Unit, 2008.
The Tuberculosis Surveillance and Research Unit (TSRU) held its last annual meeting in Helsinki, Finland, from 1 to 4 April 2008. Several topics of current interest for tuberculosis (TB) research and new research projects were presented and discussed in depth by 60 delegates from Europe, Africa and Asia. This paper summarises some of the highlights of the meeting which may be of interest to epidemiologists and managers active in the field of TB.


Revitalizing the patent system to incentivize pharmaceutical innovation: the potential of claims with means-plus-function clauses.
The pharmaceutical industry relies on innovation. However, many innovative firms are cutting their research and development investments and seeing their new product pipelines dry up, due in part to a lack of sufficient patent protection. This Note identifies two major factors that have caused this inadequacy in patent protection. First, pharmaceutical patents are challenged early and often by generic manufacturers, as encouraged by the 1984 Hatch-Waxman Act. Second, the scope of pharmaceutical-patents is sometimes unduly restrained due to limited application of the doctrine of equivalents. Consequently, pharmaceutical patents, especially drug-product patents, are easily designed around and cannot offer the protection necessary for innovative firms to recoup their developmental costs. This Note argues for a wider application of means-plus-function clauses in pharmaceutical patents as a potential cure for this problem. Means-plus-function claims, although authorized by Congress in the 1952 Patent Act, have not been explored much in the pharmaceutical context. This Note argues that this claiming strategy is not only appropriate but also particularly effective for pharmaceutical patents. Means-plus-function claims would give drug-product patents adequate scope even with the limited use of the doctrine of equivalents and thus would provide the protection necessary for innovative firms to withstand frequent attacks by generic manufacturers. Finally, this Note examines issues anticipated with applying means-plus-function claims to pharmaceutical patents and proposes possible solutions.


[Biological mechanisms of the individual variability of the anxiolytic action of opioids].
The opioid system is one in a number of peptidergic regulatory systems that support an appropriate anxiety level. The drugs that interact with the opioid system have been shown to influence anxiety, although there is a notable variability in their pharmacological effects. Biological mechanisms of this variability are considered to be the heterogeneity of opioid receptors, the ratio of the processes of their expression and desensitization, and the balance between the synthesis and degradation of endogenous opioid peptides. For instance, the anxiolythic effect of the synthetic derivate of enkephalin in rats was detected after stress-induced exhaustion of endogenous opioids, and its efficacy depended on the degree of delta- and mu-opioid receptor desensitization in some brain regions. The anxiolythic properties of the heptapeptide Selanc that has also been shown to affect the opioid system are most prominent in subjects with elevated activity of enzymes degrading endogenous opioid peptides. Thus, delicate correction of the opioid system with drugs of peptide nature is supposed to become a new approach to treatment of some forms of anxiety disorders accompanied with exhaustion of the endogenous opioid system and, in particular, of generalized anxiety disorder.


Pair-flowered cymes in the Lamiales: structure, distribution and origin.
In the Lamiales, indeterminate thyrses (made up of axillary cymes) represent a significant inflorescence type. However, it has been largely overlooked that there occur two types of cymes: (1) ordinary cymes, and (2) 'pair-flowered cymes' (PFCs), with a flower pair (terminal and front flower) topping each cyme unit. PFCs are unique to the Lamiales and their distribution, origin and phylogeny are not well understood. The Lamiales are screened as to the occurrence of PFCs, ordinary cymes and single flowers (constituting racemic inflorescences). PFCs are shown to exhibit a considerable morphological and developmental diversity and are documented to occur in four neighbouring taxa of Lamiales: Calceolariaceae, Sanango, Gesneriaceae and Plantaginaceae. They are omnipresent in the Calceolariaceae and almost so in the Gesneriaceae. In the Plantaginaceae, PFCs are restricted to the small sister tribes Russelieae and Cheloneae (while the large remainder has single flowers in the leaf/bract axils; ordinary cymes do not occur). Regarding the origin of PFCs, the inflorescences of the genus Peltanthera (unplaced as to family; sister to Calceolariaceae, Sanango and Gesneriaceae in most molecular phylogenies) support the idea that PFCs have originated from paniculate systems, with the front-flowers representing remnant flowers. From the exclusive occurrence of PFCs in the Lamiales and the proximity of the respective taxa in molecular phylogenies it may be expected that PFCs have originated once, representing a synapomorphy for this group of taxa and fading out within the Plantaginaceae. However, molecular evidence is ambiguous. Depending on the position of Peltanthera (depending in turn on the kind and number of genes and taxa analysed) a single, a double (the most probable scenario) or a triple origin appears conceivable.


Caste-specific expression patterns of immune response and chemosensory related genes in the leaf-cutting ant, Atta vollenweideri.
Leaf-cutting ants are evolutionary derived social insects with elaborated division of labor and tremendous colony sizes with millions of workers. Their social organization is mainly based on olfactory communication using different pheromones and is promoted by a pronounced size-polymorphism of workers that perform different tasks within the colony. The size polymorphism and associated behaviors are correlated to distinct antennal lobe (AL) phenotypes. Two worker phenotypes differ in number of olfactory glomeruli in the AL and the presence or absence of an extremely large glomerulus (macroglomerulus), involved in trail-pheromone reception. The males' AL contains three macroglomeruli which are presumably involved in detection of sex-pheromone components. We investigated the antennal transcriptome data of all major castes (males, queens and workers) and two worker subcastes (large and tiny workers). In order to identify putative odorant receptor genes involved in pheromone detection, we identified differentially expressed odorant receptor genes (OR-genes) using custom microarrays. In total, we found 185 OR-gene fragments that are clearly related to ORs and we identified orthologs for 70 OR-genes. Among them one OR-gene differs in relative expression between the two worker subcastes by a factor of >3 and thus is a very promising candidate gene for the trail-pheromone receptor. Using the relative expression of OR-genes in males versus queens, we identified 2 candidates for sex-pheromone receptor genes in males. In addition, we identified genes from all other chemosensory related gene families (13 chemosensory protein genes, 8 odorant binding protein genes, 2 sensory-neuron membrane protein genes, 7 ionotropic receptor genes, 2 gustatory receptor genes), and we found ant-specific expansions in the chemosensory protein gene family. In addition, a large number of genes involved in immune defense exhibited differential expression across the three different castes, and some genes even between the two worker subcastes.


Quantifying the Nonlinear Interaction in the Nervous System Based on Phase-Locked Amplitude Relationship.
This paper introduces the Cross-frequency Amplitude Transfer Function (CATF), a model-free method for quantifying nonlinear stimulus-response interaction based on phase-locked amplitude relationship. The CATF estimates the amplitude transfer from input frequencies at stimulation signal to their harmonics/intermodulation at the response signal. We first verified the performance of CATF in simulation tests with systems containing a static nonlinear function and a linear dynamic, i.e., Hammerstein and Wiener systems. We then applied the CATF to investigate the second-order nonlinear amplitude transfer in the human proprioceptive system from the periphery to the cortex. The simulation demonstrated that the CATF is a general method, which can well quantify nonlinear stimulus-response amplitude transfer for different orders of nonlinearity in Wiener or Hammerstein system configurations. Applied to the human proprioceptive system, we found a complicated nonlinear system behavior with substantial amplitude transfer from the periphery stimulation to cortical response signals in the alpha band. This complicated system behavior may be associated with the nonlinear behavior of the muscle spindle and the dynamic interaction in the thalamocortical radiation. This paper provides a new tool to identify nonlinear interaction in the nervous system. The results provide novel insight of nonlinear dynamics in the human proprioceptive system.


Major depressive disorder and nutritional medicine: a review of monotherapies and adjuvant treatments.
A literature review was conducted to examine the evidence for nutritional interventions in depression. It revealed a number of significant conclusions. Interestingly, more positive clinical trials were found to support adjuvant, rather than monotherapeutic, use of nutrients to treat depression. Much evidence exists in the area of adjuvant application of folic acid, S-adenosyl-methionine, omega-3, and L-tryptophan with antidepressants. Current evidence does not support omega-3 as an effective monotherapy to treat depression. However, this may be due, at least in part, to olive oil being used as the control intervention, some studies using docosahexaenoic acid alone or a higher docosahexaenoic acid to eicosapentaenoic acid ratio, and significant heterogeneity regarding depressive populations. Nevertheless, adjunctive prescription of omega-3 with antidepressants, or in people with dietary deficiency, may be beneficial. Inositol lacks evidence as an effective antidepressant and cannot be currently recommended. Evidence on the use of L-tryptophan for depression is inconclusive, and additional studies utilizing a more robust methodology are required.


Cryptosporidium and Giardia in marine-foraging river otters (Lontra canadensis) from the Puget Sound Georgia Basin ecosystem.
Species of Cryptosporidium and Giardia can infect humans and wildlife and have the potential to be transmitted between these 2 groups; yet, very little is known about these protozoans in marine wildlife. Feces of river otters (Lontra canadensis), a common marine wildlife species in the Puget Sound Georgia Basin, were examined for species of Cryptosporidium and Giardia to determine their role in the epidemiology of these pathogens. Using ZnSO4 flotation and immunomagnetic separation, followed by direct immunofluorescent antibody detection (IMS/DFA), we identified Cryptosporidium sp. oocysts in 9 fecal samples from 6 locations and Giardia sp. cysts in 11 fecal samples from 7 locations. The putative risk factors of proximate human population and degree of anthropogenic shoreline modification were not associated with the detection of Cryptosporidium or Giardia spp. in river otter feces. Amplification of DNA from the IMS/DFA slide scrapings was successful for 1 sample containing > 500 Cryptosporidium sp. oocysts. Sequences from the Cryptosporidium 18S rRNA and the COWP loci were most similar to the ferret Cryptosporidium sp. genotype. River otters could serve as reservoirs for Cryptosporidium and Giardia species in marine ecosystems. More work is needed to better understand the zoonotic potential of the genotypes they carry as well as their implications for river otter health.


Modulation of fluoride toxicity in rats by calcium carbonate and by withdrawal of fluoride exposure.
In order to assess the effect of calcium on the toxic effects of fluoride, adult female Wistar rats were treated with sodium fluoride (NaF, 500 ppm in drinking water) alone or in combination with calcium carbonate (CaCO3, 50 mg/ kg by oral intubation) daily for 60 days. Food, water and fluoride intake were measured daily for 60 days. Body weight gain, exploratory motor activity, rota-rod motor coordination, dental structure, activities of acetylcholinesterase (AchE, brain and skeletal muscle) and Na+ K+ ATPase (erythrocyte membrane and skeletal muscle) and the concentrations of protein (serum and skeletal muscle), calcium (serum) and fluoride (serum) were determined in these animals 24 hr after the last treatment. The same parameters were tested in another group, 60 days after withdrawal of NaF exposure (500 ppm in drinking water daily for 60 days). NaF treatment decreased food and water intake, reduced body-weight gain and impaired exploratory motor activity and rota-rod performance. Dental lesions, inhibition of the activities of AchE and N+ K+ ATPase and a decrease in the concentration of protein, and serum calcium were also observed in these animals. These effects were accompanied by a marked elevation of fluoride concentration in the serum. CaCO3 decreased the concentration of fluoride in the serum of NaF-treated animals. A decrease in serum fluoride concentration was found also after NaF withdrawal. A prevention of locomotor behavioural, biochemical and dental toxicities of fluoride was observed both in these groups. It is concluded that the dose of CaCO3 used in the present study has a potential to prevent the toxicity of fluoride by maintaining serum fluoride at a less toxic level. Further, the toxic effects of fluoride are reversible if its exposure is withdrawn for 2 months.


Firing patterns and synaptic potentials of identified giant aspiny interneurons in the rat neostriatum.
Intracellular recordings were made in vivo from 9 giant aspiny neurons in the neostriatum of urethane-anesthetized rats. The cells were identified by intracellular staining with HRP or biocytin. The neurons exhibited morphological features typical of neostriatal cholinergic interneurons. Six of the cells were obtained from intact animals, while 3 were recorded from rats with ipsilateral hemidecortications. Giant aspiny neurons were characterized by their slow irregular but tonic (3-10/sec) spontaneous activity and long-duration action potentials. Examination of the underlying membrane potential trajectories during spontaneous firing revealed that individual action potentials were triggered from spontaneous small (1-5 mV) depolarizing potentials. These spontaneous potentials exhibited the voltage sensitivity of ordinary EPSPs. They were much less frequent during the 80-200 msec pause in tonic afferent input that follows the excitation evoked by cortical or thalamic stimulation, and were decreased in frequency in decorticate animals. Their rise times and half-widths matched those expected for unitary synaptic potentials placed proximally on the surface of the neurons. Low-intensity stimulation of neostriatal afferents produced small short-latency EPSPs that appeared to be composed of responses identical to the spontaneous depolarizing potentials. The latencies of the EPSPs evoked from the cerebral cortex and thalamus were consistent with a monosynaptic input from both structures, but the maximal size of the EPSPs was much smaller than that evoked in spiny neurons, suggesting that a smaller number of afferent inputs make synapses with each of the aspiny cells. Giant aspiny neurons exhibited much larger input resistances and longer time constants than spiny neostriatal neurons. They also exhibited relatively linear steady-state current-voltage relationship compared to spiny projection cells. Input resistances ranged from 71-105 M omega, and time constants ranged from 17.8-28.5 msec. Analysis of the charging transients in response to current pulses yielded estimates of dendritic length of approximately 1 length constant. Repetitive firing of the neurons was limited by a powerful spike afterhyperpolarization and by a strong spike frequency adaptation. The sensitivity of the giant aspiny interneuron to a relatively small number of proximal afferent synaptic contacts, its tonic firing, and its widespread dendritic and axonal fields place it in an excellent position to act as a modulator of the excitability of neostriatal projection neurons in advance of the onset of movement-related neostriatal activity.


Sports Injury Forecasting and Complexity: A Synergetic Approach.
The understanding that sports injury is the result of the interaction among many factors and that specific profiles could increase the risk of the occurrence of a given injury was a significant step in establishing programs for injury prevention. However, injury forecasting is far from being attained. To be able to estimate future states of a complex system (forecasting), it is necessary to understand its nature and comply with the methods usually used to analyze such a system. In this sense, sports injury forecasting must implement the concepts and tools used to study the behavior of self-organizing systems, since it is by self-organizing that systems (i.e., athletes) evolve and adapt (or not) to a constantly changing environment. Instead of concentrating on the identification of factors related to the injury occurrence (i.e., risk factors), a complex systems approach looks for the high-order variables (order parameters) that describe the macroscopic dynamic behavior of the athlete. The time evolution of this order parameter informs on the state of the athlete and may warn about upcoming events, such as injury. In this article, we describe the fundamental concepts related to complexity based on physical principles of self-organization and the consequence of accepting sports injury as a complex phenomenon. In the end, we will present the four steps necessary to formulate a synergetics approach based on self-organization and phase transition to sports injuries. Future studies based on this experimental paradigm may help sports professionals to forecast sports injuries occurrence.


DNA barcodes from century-old type specimens using next-generation sequencing.
Type specimens have high scientific importance because they provide the only certain connection between the application of a Linnean name and a physical specimen. Many other individuals may have been identified as a particular species, but their linkage to the taxon concept is inferential. Because type specimens are often more than a century old and have experienced conditions unfavourable for DNA preservation, success in sequence recovery has been uncertain. This study addresses this challenge by employing next-generation sequencing (NGS) to recover sequences for the barcode region of the cytochrome c oxidase 1 gene from small amounts of template DNA. DNA quality was first screened in more than 1800 century-old type specimens of Lepidoptera by attempting to recover 164-bp and 94-bp reads via Sanger sequencing. This analysis permitted the assignment of each specimen to one of three DNA quality categories--high (164-bp sequence), medium (94-bp sequence) or low (no sequence). Ten specimens from each category were subsequently analysed via a PCR-based NGS protocol requiring very little template DNA. It recovered sequence information from all specimens with average read lengths ranging from 458 bp to 610 bp for the three DNA categories. By sequencing ten specimens in each NGS run, costs were similar to Sanger analysis. Future increases in the number of specimens processed in each run promise substantial reductions in cost, making it possible to anticipate a future where barcode sequences are available from most type specimens.


Nitric oxide stimulates gamma-aminobutyric acid release and inhibits glycine release in retina.
Nitric oxide (NO) modulates the uptake and/or release of neurotransmitters through a variety of cellular mechanisms. However, the pharmacological and biochemical processes underlying these neurochemical effects of NO often remain unclear. In our study, we used immunocytochemical methods to study the effects of NO, cyclic guanosine monophosphate (cGMP), and peroxynitrite on the uptake and release of gamma-aminobutyric acid (GABA) and glycine in the turtle retina. In addition, we examined the involvement of glutamate receptors, calcium, and the GABA transporter in this GABA uptake and release. We also tested for interactions between the GABAergic and glycinergic systems. In general, we show that NO stimulated GABA release and inhibited glycine release. The NO-stimulated GABA release involved calcium-dependent or calcium-independent synaptic release or reversal of the GABA transporter. Some effects of NO on GABA release involved glutamate, cGMP, or peroxynitrite. NO promoted glycine uptake and inhibited its release, and this inhibition of glycine release was influenced by GABAergic modulation. These findings indicate that NO modulates the levels of the inhibitory transmitters GABA and glycine through several specific biochemical mechanisms in different retinal cell types and layers. Thus it appears that some of the previously described reciprocal interactions between GABA and glycine in the retina function through specific NO signaling pathways.


Gain-of-function Pyrin mutations induce NLRP3 protein-independent interleukin-1β activation and severe autoinflammation in mice.
Missense mutations in the C-terminal B30.2 domain of pyrin cause familial Mediterranean fever (FMF), the most common Mendelian autoinflammatory disease. However, it remains controversial as to whether FMF is due to the loss of an inhibitor of inflammation or to the activity of a proinflammatory molecule. We generated both pyrin-deficient mice and "knockin" mice harboring mutant human B30.2 domains. Homozygous knockin, but not pyrin-deficient, mice exhibited spontaneous bone marrow-dependent inflammation similar to but more severe than human FMF. Caspase-1 was constitutively activated in knockin macrophages and active IL-1β was secreted when stimulated with lipopolysaccharide alone, which is also observed in FMF patients. The inflammatory phenotype of knockin mice was completely ablated by crossing with IL-1 receptor-deficient or adaptor molecule ASC-deficient mice, but not NLRP3-deficient mice. Thus, our data provide evidence for an ASC-dependent NLRP3-independent inflammasome in which gain-of-function pyrin mutations cause autoinflammatory disease.


Kinetoplastid flagellates overlooked by universal primers dominate in the oxygenated hypolimnion of Lake Biwa, Japan.
Kinetoplastid flagellates, microscopically often detected from various aquatic environments and considered ubiquitous are seldom reported in molecular diversity studies with universal eukaryote DNA primers. To investigate this inconsistency, we examined nanoflagellate diversity in Lake Biwa, Japan by 18S rRNA gene clone libraries using universal eukaryote and kinetoplastid-specific primers. We also examined the abundance of kinetoplastids by Catalyzed Reporter Deposition-Fluorescence In Situ Hybridization. No, kinetoplastid sequences were detected in the universal eukaryote primers library from epilimnion and hypolimnion in different seasons. However, kinetoplastid flagellates were detected with kinetoplastid-specific probe from all of the seasons and contributed up to 11.9 and 36.0% of total eukaryotes in the epilimnion and hypolimnion, respectively. Thus, kinetoplastids probably are a significant, sometimes dominant, group in the hypolimnion, contributing up to 43.7% of the total flagellates. Using group-specific primers, kinetoplastid sequences were also obtained from both epilimnion and hypolimnion library. Therefore, we attributed the inconsistency to the divergent nature of 18S rRNA gene of kinetoplastids, which lead to their undetection in the universal eukaryote primer libraries. This study revealed that kinetoplastids have significant ecological importance in the hypolimnion of Lake Biwa, suggesting that these flagellates have been overlooked in other studies using universal eukaryote primers.


Retargeting of macroH2A following mitosis to cytogenetic-scale heterochromatic domains.
The heritability of chromatin states through cell division is a potential contributor to the epigenetic maintenance of cellular memory of prior states. The macroH2A histone variant has properties of a regulator of epigenetic cell memory, including roles controlling gene silencing and cell differentiation. Its mechanisms of regional genomic targeting and maintenance through cell division are unknown. Here, we combined in vivo imaging with biochemical and genomic approaches to show that human macroH2A is incorporated into chromatin in the G1 phase of the cell cycle following DNA replication. The newly incorporated macroH2A retargets the same large heterochromatic domains where macroH2A was already enriched in the previous cell cycle. It remains heterotypic, targeting individual nucleosomes that do not already contain a macroH2A molecule. The pattern observed resembles that of a new deposition of centromeric histone variants during the cell cycle, indicating mechanistic similarities for macrodomain-scale regulation of epigenetic properties of the cell.


Presence of Rickettsia Species in a Marginalized Area of Yucatan, Mexico.
In the state of Yucatan, Mexico, rickettsiosis has become a common vector-borne disease in the general population. Ectoparasite species such as Rhipicephalus sanguineus and Amblyomma mixtum have been identified as Rickettsia vectors in Yucatan by studies focused on the wild animal population in rural areas. There have been studies that have tried to determine the presence of Rickettsia species in ectoparasites collected in Yucatan, but these studies did not include marginalized areas, where living in close contact with domestic and peridomestic animals that carry ectoparasites is a high-risk factor for acquiring rickettsial infection or many other vector-borne diseases. We evaluated the vector diversity and the presence of Rickettsia species presence in the ectoparasite population that parasitizes domestic animals in a marginalized rural town of Yucatan, Mexico; we also evaluated the seroprevalence of rickettsial antibodies in the human population of this town in order to determine the prevalence of rickettsial infection. A total of 437 ectoparasites were collected from the study area. The tick specimens collected belonged to the species Rhipicephalus sanguineus (n=380, 49 positive), Amblyomma mixtum (n=3, 0 positive), Ixodes affinis (n=4, 0 positive), Ctenocephalides felis (n=33, 0 positive), and Trichodectes canis (n=17, 0 positive). Conventional polymerase chain reaction and sequencing were used to identify the DNA of Rickettsia. Six out of 354 (1.8%) serum samples were positive for antibody to R. typhi. The combination of low antibody titers and the presence of Rickettsia species infecting ectoparasite species found in the study area requires eco-epidemiological studies and the identification of potentially protective practices or habits.


Mindfulness in people with a respiratory diagnosis: A systematic review.
To describe how mindfulness is delivered and to examine the effect of mindfulness on health-related quality of life (HRQOL), mindful awareness and stress in adults with a respiratory diagnosis. Five electronic databases were searched. Data were extracted and assessed for quality by two reviewers. Data were extracted from four studies. Interventions were based on Mindfulness-Based Stress Reduction and delivered by trained instructors. Recordings of mindfulness were provided for home-based practice. One study targeted the intervention exclusively to anxious individuals with a respiratory diagnosis. Adherence to mindfulness was poor. No effects were seen on disease-specific HRQOL (standardized mean difference (SMD)=-0.21 95% CI: -0.36 to 0.48, p=0.78), mindful awareness (SMD=0.09 95% CI: -0.34 to 0.52, p=0.68) or stress levels (SMD =-0.11 95% CI: -0.46 to 0.23, p=0.51). Mindfulness interventions, delivered to individuals with a respiratory diagnosis, varied widely in terms of delivery and the outcomes assessed making it difficult to draw any conclusions regarding its effectiveness.


Climate change and local public health in the United States: preparedness, programs and perceptions of local public health department directors.
While climate change is inherently a global problem, its public health impacts will be experienced most acutely at the local and regional level, with some jurisdictions likely to be more burdened than others. The public health infrastructure in the U.S. is organized largely as an interlocking set of public agencies at the federal, state and local level, with lead responsibility for each city or county often residing at the local level. To understand how directors of local public health departments view and are responding to climate change as a public health issue, we conducted a telephone survey with 133 randomly selected local health department directors, representing a 61% response rate. A majority of respondents perceived climate change to be a problem in their jurisdiction, a problem they viewed as likely to become more common or severe over the next 20 years. Only a small minority of respondents, however, had yet made climate change adaptation or prevention a top priority for their health department. This discrepancy between problem recognition and programmatic responses may be due, in part, to several factors: most respondents felt personnel in their health department--and other key stakeholders in their community--had a lack of knowledge about climate change; relatively few respondents felt their own health department, their state health department, or the Centers for Disease Control and Prevention had the necessary expertise to help them create an effective mitigation or adaptation plan for their jurisdiction; and most respondents felt that their health department needed additional funding, staff and staff training to respond effectively to climate change. These data make clear that climate change adaptation and prevention are not currently major activities at most health departments, and that most, if not all, local health departments will require assistance in making this transition. We conclude by making the case that, through their words and actions, local health departments and their staff can and should play a role in alerting members of their community about the prospect of public health impacts from climate change in their jurisdiction.


Night shift work and cardiovascular disease biomarkers in female nurses.
Night shift work is associated with cardiovascular disease, but its associations with cardiovascular disease biomarkers are unclear. We investigated these associations in a study of female nurses. We used data from the Nurses' Health Study II for total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein (HDL) cholesterol, triglycerides, C-reactive protein (CRP), and fibrinogen. The sample sizes for our analysis ranged from 458 (fibrinogen) to 3574 (total cholesterol). From questionnaires, we determined the number of night shifts worked in the 2 weeks before blood collection and total years of rotating night shift work. We used quantile regression to estimate differences in biomarker levels by shift work history, adjusting for potential confounders. Nurses working 1 to 4 recent night shifts had median HDL cholesterol levels 4.4 mg/dL (95% confidence interval [CI]: 0.3, 7.5) lower than nurses without recent night shifts. However, working ≥5 recent night shifts and years of rotating night shift work were not associated with HDL cholesterol. There was no association between recent night shifts and CRP, but median CRP levels were 0.1 (95% CI: 0.0, 0.2), 0.2 (95% CI: 0.1, 0.4), and 0.2 (95% CI: 0.0, 0.4) mg/L higher among nurses working rotating night shifts for 1 to 5, 6 to 9, and ≥10 years compared with nurses never working rotating night shifts. These associations were attenuated when excluding postmenopausal women and women taking statins. We observed no associations between night shift work and other biomarkers. We found suggestive evidence of adverse short-term and long-term effects of night shift work on select cardiovascular disease biomarkers.


MicroRNA-188 regulates age-related switch between osteoblast and adipocyte differentiation.
Bone marrow mesenchymal stem cells (BMSCs) exhibit an age-dependent reduction in osteogenesis that is accompanied by an increased propensity toward adipocyte differentiation. This switch increases adipocyte numbers and decreases the number of osteoblasts, contributing to age-related bone loss. Here, we found that the level of microRNA-188 (miR-188) is markedly higher in BMSCs from aged compared with young mice and humans. Compared with control mice, animals lacking miR-188 showed a substantial reduction of age-associated bone loss and fat accumulation in bone marrow. Conversely, mice with transgenic overexpression of miR-188 in osterix+ osteoprogenitors had greater age-associated bone loss and fat accumulation in bone marrow relative to WT mice. Moreover, using an aptamer delivery system, we found that BMSC-specific overexpression of miR-188 in mice reduced bone formation and increased bone marrow fat accumulation. We identified histone deacetylase 9 (HDAC9) and RPTOR-independent companion of MTOR complex 2 (RICTOR) as the direct targets of miR-188. Notably, BMSC-specific inhibition of miR-188 by intra-bone marrow injection of aptamer-antagomiR-188 increased bone formation and decreased bone marrow fat accumulation in aged mice. Together, our results indicate that miR-188 is a key regulator of the age-related switch between osteogenesis and adipogenesis of BMSCs and may represent a potential therapeutic target for age-related bone loss.


Role of the thalamus in natural recovery of cognitive impairment in patients with mild traumatic brain injury.
Patients with mild traumatic brain injury (mTBI) may have normal neuroimaging but manifest with a broad-spectrum of cognitive-deficits, which may resolve eventually. The function of the thalamus in the process of natural-recovery remains elusive. The current study investigates the role of the thalamus in natural-recovery of cognitive-deficits in patients with mTBI. Twenty-one patients with mTBI were evaluated with an initial MRI scan, within 36 hours of injury and assessed with neuropsychological tests(NPT) at 3-4 weeks after injury. First and second follow-up MRI and NPT were performed at 3-4 months and 6-7 months, respectively. The volume and tensor measures of the thalamus and cognitive-scores were analysed at each assessment using repeated-measures of variance. The association of cognitive-scores with corresponding period imaging measures was analysed using bivariate-correlation. Serial evaluation showed that all the cognitive-domains improved significantly. During this period there was a significant increase in mean thalamic volume (p = 0.049, effect-size = 0.18). After 3-4 months there was emergence of anisotropic thalamo-cortical connections. At 2-3 weeks and 6-7 months after injury, the alterations in diffusivity values were positively associated with improvement in memory-scores. Improvement in attention-scores correlated significantly with changes in tensor values at the 6-7 months after-injury. The correlation between improvement in cognitive-scores and changes in thalamic tensor and volume measures reflect the role of the thalamus in natural-recovery after mTBI.


Evaluation of postoperative pregabalin for attenuation of postoperative shoulder pain after thoracotomy in patients with lung cancer, a preliminary result.
Thirty-one to 97% of patients who undergo thoracotomy for lung cancer experience ipsilateral shoulder pain, marring the otherwise excellent relief provided by thoracic epidural analgesia. The aim of this study was to test whether the addition of pregabalin to the treatment for shoulder pain would provide a significant benefit. Twenty patients undergoing thoracic surgery for lung cancer were enrolled in the control group between May 2012 and December 2012, and 20 patients were enrolled in the pregabalin group between January 2013 and July 2013, consecutively. All patients had standard pre- and intraoperative care. Patients received pregabalin 150 mg po POD 1 and then non-steroidal anti-inflammatory drugs (NSAIDs) po 2 h later (pregabalin group), or they received only NSAIDs po at exactly the same times (control group). Pain severity was then measured using a 100-mm visual analog scale (VAS) scoring system. The VAS scores indicated that patients in the pregabalin group had significantly less shoulder pain on postoperative day (POD) 2 than those in the control group (control: 27.9 ± 28.1 vs. pregabalin: 11.8 ± 14.4; p = 0.030). No differences in pain were observed between the two groups on other POD. There were significant differences on only POD 2 in the patients with shoulder pain immediately after surgery. Three of the pregabalin-treated patients showed mild somnolence. Postoperative administration of pregabalin provided significant relief of postoperative shoulder pain during earlier POD after thoracic surgery for lung cancer when received multimodal analgesia in combination with NSAIDs.


Daily minority stress and affect among gay and bisexual men: A 30-day diary study.
This study examined the time-variant association between daily minority stress and daily affect among gay and bisexual men. Tests of time-lagged associations allow for a stronger causal examination of minority stress-affect associations compared with static assessments. Multilevel modeling allows for comparison of associations between minority stress and daily affect when minority stress is modeled as a between-person factor and a within-person time-fluctuating state. 371 gay and bisexual men in New York City completed a 30-day daily diary, recording daily experiences of minority stress and positive affect (PA), negative affect (NA), and anxious affect (AA). Multilevel analyses examined associations between minority stress and affect in both same-day and time-lagged analyses, with minority stress assessed as both a between-person factor and a within-person state. Daily minority stress, modeled as both a between-person and within-person construct, significantly predicted lower PA and higher NA and AA. Daily minority stress also predicted lower subsequent-day PA and higher subsequent-day NA and AA. Self-report assessments and the unique sample may limit generalizability of this study. The time-variant association between sexual minority stress and affect found here substantiates the basic tenet of minority stress theory with a fine-grained analysis of gay and bisexual men's daily experience. Time-lagged effects suggest a potentially causal pathway between minority stress as a social determinant of mood and anxiety disorder symptoms among gay and bisexual men. When modeled as both a between-person factor and within-person state, minority stress demonstrated expected patterns with affect.


Podoplanin expression is a prognostic biomarker but may be dispensable for the malignancy of glioblastoma.
Treatment options of glioblastoma, the most aggressive primary brain tumor with frequent relapses and high mortality, are still very limited, urgently calling for novel therapeutic targets. Expression of the glycoprotein podoplanin correlates with poor prognosis in various cancer entities, including glioblastoma. Furthermore, podoplanin has been associated with tumor cell migration and proliferation in vitro; however, experimental data on its function in gliomagenesis in vivo are still missing. Hence, we have functionally investigated the impact of podoplanin on glioblastoma in a preclinical mouse model to evaluate its potential as a therapeutic target. Fluorescence activated cell sorting, genome-wide expression analysis, and clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9)-mediated deletion of podoplanin in patient-derived human glioblastoma cells were combined with organotypic brain slice cultures and intracranial injections into mice. We defined a malignant gene signature in tumor cells with high podoplanin expression. The increase and/or maintenance of high podoplanin expression in serial transplantations and in podoplaninlow-sorted glioblastoma cells during outgrowth indicated the association of high podoplanin expression and poor outcome. Unexpectedly, similar rates of proliferation, apoptosis, angiogenesis, and invasion were observed in control and podoplanin-deleted tumors. Accordingly, neither tumor growth nor survival was affected upon podoplanin loss. We report that tumor progression occurs independently of podoplanin. Thus, in contrast to previous suggestions, blocking of podoplanin does not represent a promising therapeutic approach. However, as podoplanin is associated with tumor aggressiveness and progression, we propose the cell surface protein as a biomarker for poor prognosis.


Simplified cephalometric lines for the estimation of muscular lines of action.
In the study of masticatory muscle performance, one of the biomechanical variables that can be estimated is the mechanical advantage of the masticatory muscles, namely, the ratio between the muscular moment arm and the bite force moment arm. In the present study, the position of the estimated line of action of the masseter muscle, drawn between gonion and orbitale (Go-Or) relative to dental (occlusal plane) and skeletal (Frankfort plane) references was analyzed in 431 pretreatment lateral cephalograms of orthodontic patients (195 males, 236 females) aged 6 to 50 years, and in the lateral tracings of the Bolton standards (6 to 18 years of age). The following measurements were evaluated: (1) skeletocutaneous class (soft tissue equivalent of Wits appraisal, linear distance in millimeters between the projections of points A' and B' on the occlusal plane); (2) angle between the Go-Or line and the perpendicular to the occlusal plane at the molar occlusal point; and (3) angle between the Go-Or line and the Frankfort plane. In the patients, the skeletocutaneous class ranged between--14.5 and 15.5 mm, without any sex- or age-related differences. The angle between the Go-Or line and the perpendicular to the occlusal plane was, on average, 39 degrees (range 15 to 53 degrees), and it decreased with advancing age; while the average angle between Go-Or and Frankfort plane was 42 degrees (range 30 to 54 degrees), and it increased in older patients. No effects of sex were found. The two angles were significantly correlated to each other, while no correlations were found with the sagittal jaw discrepancy. Similar results were obtained on the Bolton tracings. Overall, the present cephalometric analogue could be useful in biomechanical simulations.


Asthma and selective migration from farming environments in a three-generation cohort study.
Individuals raised on a farm appear to have less asthma than individual raised elsewhere. However, selective migration might contribute to this as may also the suggested protection from farm environment. This study investigated if parents with asthma are less likely to raise their children on a farm. This study involved three generations: 6045 participants in ECRHS/RHINE cohorts (born 1945-1973, denoted G1), their 10,121 parents (denoted G0) and their 8260 offspring participating in RHINESSA (born 1963-1998, denoted G2). G2-offspring provided information on parents not participating in ECRHS/RHINE. Asthma status and place of upbringing for all three generations were reported in questionnaires by G1 in 2010-2012 and by G2 in 2013-2016. Binary regressions with farm upbringing as outcome were performed to explore associations between parental asthma and offspring farm upbringing in G0-G1 and G1-G2. Having at least one parent with asthma was not associated with offspring farm upbringing, either in G1-G2 (RR 1.11, 95% CI 0.81-1.52) or in G0-G1 (RR 0.99, 0.85-1.15). G1 parents with asthma born in a city tended to move and raise their G2 offspring on a farm (RR 2.00, 1.12-3.55), while G1 parents with asthma born on a farm were less likely to raise their G2 offspring on a farm (RR 0.34, 0.11-1.06). This pattern was not observed in analyses of G0-G1. This study suggests that the protective effect from farm upbringing on subsequent asthma development could not be explained by selective migration. Intriguingly, asthmatic parents appeared to change environment when having children.


Etiology of acute gastroenteritis in children requiring hospitalization in the Netherlands.
Infectious gastroenteritis causes a considerable burden of disease worldwide. Costs due to gastroenteritis are dominated by the hospitalized cases. Effective control of gastroenteritis should be targeted at the diseases with the highest burden and costs. For that, an accurate understanding of the relative importance of the different bacterial, viral, and parasitic pathogens is needed. The objective of the present study was to determine the incidence and etiology of gastroenteritis requiring hospital admission in the Netherlands. Six hospitals enrolled patients admitted with gastroenteritis for approximately one year over the period May 2008 to November 2009. Participants provided questionnaires and a fecal sample, and the hospital filled out a clinical questionnaire. In total, 143 children hospitalized for gastroenteritis and 64 matched controls were included in the study. Overall incidence of gastroenteritis requiring hospitalization was estimated at 2.92 per 1,000 children aged 0-17 years per year, with the highest incidence in children under the age of 5 years. The full diagnostic panel of pathogens could be studied in fecal samples of 96 cases. One or more pathogens were found in 98% of these cases. Co-infections were observed relatively often (40%). Viruses were detected in 82% of the samples, with rotavirus being most common (56%), bacteria in 32% and parasites in 10%. The present study emphasizes the importance of viral pathogens, especially rotavirus, in hospitalizations of children with gastroenteritis. Policies to reduce (costs of) hospitalizations due to gastroenteritis should therefore be first targeted at rotavirus.


Treatment for hyperactive children: homeopathy and methylphenidate compared in a family setting.
The sharp increase of the prescription of methylphenidate (MPD) in hyperactive children in recent years is a matter of increasing uneasiness among professionals, parents and politicians. There is little awareness of treatment alternatives. The purpose of this prospective trial was to assess the efficacy of homeopathy in hyperactive patients and to compare it MPD. The study was performed in a paediatric practice with conventional and homeopathic backgrounds. Children aged 3-17 y, conforming to the DSM-IV criteria for attention deficit hyperactivity disorder (ADHD) with a Conners Global Index (CGI) of 14 or higher were eligible for the study. All of them received an individual homeopathic treatment. When clinical improvement reached 50%, the parents were asked to reevaluate the symptoms. Those who did not improve sufficiently on homeopathy were changed to MPD, and again evaluated after 3 months. One hundred and fifteen children (92 boys, 23 girls) with a mean age of 8.3 y at diagnosis were included in the study. Prior to treatment the mean CGI was 20.63 (14-30), the mean index of the homeopathy group 20.52 and of the MPD-group 20.94. After an average treatment time of 3.5 months 86 children (75%) had responded to homeopathy, reaching a clinical improvement rating of 73% and an amelioration of the CGI of 55%. Twenty-five children (22%) needed MPD; the average duration of homeopathic (pre-) treatment in this group was 22 months. Clinical improvement under MPD reached 65%, the lowering of the CGI 48%. Three children did not respond to homeopathy nor to MPD, and one left the study. In cases where treatment of a hyperactive child is not urgent, homeopathy is a valuable alternative to MPD. The reported results of homeopathic treatment appear to be similar to the effects of MPD. Only children who did not reach the high level of sensory integration for school had to be changed to MPD. In preschoolers, homeopathy appears a particularly useful treatment for ADHD.


Auditory middle-latency responses in humans.
Middle-latency responses (MLR) in humans were studied using an unconventional recording technique with wide bandpass filters. Such filtering permitted simultaneous recording of the auditory brain stem response (ABR) thus facilitating comparisons between the two responses. Effects of sedation (chloral hydrate and diazepam), stimulus-related properties and the coronal distribution of MLRs were examined. Mild sedatives did not appear to affect either MLRs or ABRs. MLRs differed from ABRs in their stimulus-related properties, implying that the neuronal mechanisms underlying their generation are not the same. The amplitude of the MLR component, Pa, was largest at the vertex and symmetrically distributed over the temporal lobes. MLR components Na and Pa and ABR wave V were reliably obtained in all subjects at moderate and high stimulus intensities. At low stimulus levels, however, the detectability of wave V was more robust than the middle-latency components. Thus ABR appears to be the test of choice when hearing sensitivity is in question. MLRs are likely to be most clinically useful in patients with neurological or central auditory processing disorders.


Endoscopic Stenting Does Not Worsen Long Term Outcomes Amongst Patients Presenting with Obstruction from Colorectal Cancers.
Stenting has been increasingly adopted in colorectal cancer patients presenting with acute large bowel obstruction. However, long-term outcomes of stenting are lacking in the literature. Our study attempts to compare the long-term outcomes of colonic stenting and emergency surgery amongst left-sided colorectal cancer patients presenting with acute large bowel obstruction. A retrospective review of all patients who presented with nonmetastatic colorectal cancer who underwent either endoscopic stenting or emergency surgery for acute large bowel obstruction was performed from January 2007 to April 2016. Patients were analysed in an intention-to-treat analysis. Forty-seven (46.1%) patients underwent emergency surgery, whereas 55 (53.9%) underwent colonic stenting with a technical success rate of 71.0%. Patients who underwent emergency surgery were more likely to develop severe complications compared with patients who underwent successful colonic stenting, but the difference was not statistically significant (odds ratio [OR] 2.84; 95% confidence interval [CI] 0.71-11.3, p = 0.139). Patients were followed up for a median of 48.3 months (3.1-111) in the stenting group and 51.2 months (1.2-117.1) in the emergency surgery group. Recurrence rates between colonic stenting and emergency surgery were similar (25.6% vs. 21.3%, p = 0.500), with more anastomotic and peritoneal recurrences were noted in the emergency surgery group. 5 year disease free survival (77% vs. 73%, p = 0.708) and overall survival (86% vs. 62%, p = 0.064) also were similar. Patients who underwent endoscopic stenting for large bowel obstruction have comparable long-term outcomes as those who undergo emergency surgery. The role of endoscopic stenting in obstructed colorectal cancers merits further evaluation.


Temperature affects expression of symptoms induced by soybean mosaic virus in homozygous and heterozygous plants.
Seven strains (G1 to G7) of soybean mosaic virus (SMV) and 3 resistance loci (Rsv1, Rsv3, and Rsv4) have been identified in soybean. The interaction of SMV strains and host resistance genes results in resistant (symptomless), susceptible (mosaic), or necrotic (leaf and stem necrosis) reactions. The necrotic reaction may be gene dosage dependent and influenced by temperature. Using a set of soybean isolines and hybrids containing homozygous or heterozygous alleles of rsv, Rsv1, Rsv1-n, Rsv3, or Rsv4, this study has explored the relationship of SMV-induced symptoms and resistance gene dosage at different temperatures. Results showed that SMV-inoculated plants carrying Rsv3 or Rsv4 were symptomless at both homozygous and heterozygous states at all temperature regimes. Threshold temperatures for symptoms changing from stem tip necrosis (STN) to mosaic were 30, 33, and 33 degrees C in G7-inoculated homozygous genotypes V94-3971(Rsv1) and PI 96983 (Rsv1) and G1-inoculated V262 (Rsv1-n), respectively. However, at the heterozygous state, threshold temperature was 30 degrees C in G7-inoculated V94-3971 x Essex F(1) for the symptom change from STN to mosaic, 31 degrees C in G7-inoculated Essex x PI 96983 F(1) from STN to mixture of necrosis and mosaic (N-M), and 32 degrees C in G1-inoculated V262 x Essex F(1) from N-M to mosaic. Incomplete necrosis was observed in the heterozygous state in G1-inoculated V262 x Essex F(1) and G7-inoculated PI 96983 x Essex F(1) where necrotic and mosaic symptoms were mixed. High temperature (37 degrees C) tends to mask the expression of mosaic symptoms in both homozygous and heterozygous plants. STN expression in response to temperature was affected by resistance gene, gene dosage, host genetic background, and specific SMV strains. Thus, Rsv3 and Rsv4 are a better choice as source of genetic resistance for breeding SMV-resistant cultivars.


Spectrophotometry assays to determine G6PD activity from Trinity Biotech and Pointe Scientific G6PD show good correlation.
Spectrophotometry kits from Pointe Scientific (PS; USA) were compared to kits from Trinity Biotech (Trinity; Ireland) in 50 venous blood samples from purposively selected individuals in Bangladesh. Repeatability and inter-assay variability were assessed by Students t-test, Bland-Altman plot and Pearson correlation coefficient (r). The median glucose-6-phosphate dehydrogenase (G6PD) activity of all G6PD normal participants was calculated per assay and defined as 100% activity. Performance was calculated considering 30% and 70% cut off activities and Trinity as reference. The intra-assay correlation of Trinity (r = 0.9841, p < 0.001) and PS (r = 0.9833, p < 0.001) did not differ significantly (p = 0.904). Both assays were closely correlated (r = 0.9799, p < 0.001), with a mean difference of 0.1 U/gHb (95% limit of agreement: - 1.32 to 1.57). At 30% cut off PS had a sensitivity of 100% (95% confidence interval (95 CI) 59.0-100.0) and specificity of 100% (95% CI 91.8 to 100.0), at 70% cut-off of 100% (95% CI 79.4-100.0) and 97.1% (95% CI 84.7-99.9) respectively. The G6PD assay from PS is a reliable alternative to the assay from Trinity.


Genomic alterations in BCL2L1 and DLC1 contribute to drug sensitivity in gastric cancer.
Gastric cancer (GC) is the third leading cause of cancer-related deaths worldwide. Recent high-throughput analyses of genomic alterations revealed several driver genes and altered pathways in GC. However, therapeutic applications from genomic data are limited, largely as a result of the lack of druggable molecular targets and preclinical models for drug selection. To identify new therapeutic targets for GC, we performed array comparative genomic hybridization (aCGH) of DNA from 103 patients with GC for copy number alteration (CNA) analysis, and whole-exome sequencing from 55 GCs from the same patients for mutation profiling. Pathway analysis showed recurrent alterations in the Wnt signaling [APC, CTNNB1, and DLC1 (deleted in liver cancer 1)], ErbB signaling (ERBB2, PIK3CA, and KRAS), and p53 signaling/apoptosis [TP53 and BCL2L1 (BCL2-like 1)] pathways. In 18.4% of GC cases (19/103), amplification of the antiapoptotic gene BCL2L1 was observed, and subsequently a BCL2L1 inhibitor was shown to markedly decrease cell viability in BCL2L1-amplified cell lines and in similarly altered patient-derived GC xenografts, especially when combined with other chemotherapeutic agents. In 10.9% of cases (6/55), mutations in DLC1 were found and were also shown to confer a growth advantage for these cells via activation of Rho-ROCK signaling, rendering these cells more susceptible to a ROCK inhibitor. Taken together, our study implicates BCL2L1 and DLC1 as potential druggable targets for specific subsets of GC cases.


The international registry on hand and composite tissue transplantation.
Since May 2002, all groups performing hand transplantations have supplied information to the International Registry on Hand and Composite Tissue Transplantation (IRHCCT). The analysis of all cases with follow-up information up to September 2007 is presented here. From September 1998 to September 2007, 38 hands (18 unilateral and 10 bilateral hand transplantations) and two digits have been reported to IRHCTT, for a total of 30 patients with a follow-up period ranging from 6 months to 9 years. They were 28 males and two females, median age of 34 years. Time since hand loss ranged from 2 months to 34 years and in 50% of cases the level of amputation was at wrist level. Immunosuppressive therapy included tacrolimus, mycophenolate mofetil, rapamycin, and steroids; polyclonal or monoclonal antibodies were used for induction. Topical immunosuppression was also employed in several cases. Patient survival was 100%. Graft survival was 100% at 1 and 2 years. Except for the Chinese patients, current graft survival is 95.6% and cause of graft loss was always no compliance to the treatment. Acute rejection episodes occurred in 85% of the patients within the first year and they were reversible in all compliant patients. Side-effects included opportunistic infections and metabolic complications. All patients developed protective sensibility, 90% of them developed tactile sensibility, and 72% also a discriminative sensibility. Motor recovery enabled patients to perform most daily activities. Hand transplantation is feasible with a high success rate and satisfactory functional outcome.


X-linked immunodeficiency affects the outcome of Schistosoma mansoni infection in the murine model.
The incidence of the X-linked immunodeficiency (Xid) on the outcome of Schistosoma mansoni infection has been evaluated through a comparative analysis of parasitological and immune parameters in two different mouse strains: control BALB/c and BALB. Xid mice which carry the Xid mutation and lack B1 (CD5+ B) cells. This study clearly demonstrates that infected B1 cell-deficient animals display a higher susceptibility to S. mansoni infection as revealed by an increase in the tissue egg loads and a significantly elevated mortality, as well as an increase in the granuloma densities. The analysis of the humoral and the cellular responses, conducted in the same experimental conditions, indicates differences in terms of cytokine production after specific antigenic stimulation of splenocytes. Larger amounts of IFN-gamma and IL-4 are observed in BALB. Xid mice while IL-10 production is reduced. In parallel, the study of the specific antibody isotype profiles shows higher amounts of specific IgE and IgG1 antibodies and lower amounts of IgM and IgA in BALB. Xid mice. Taken together, these observations support the idea that B cells are playing a role in the ability of mice to tolerate infection with Schistosoma mansoni.


[Endemic pemphigus foliaceus in a pregnant woman. Report of one case].
Endemic pemphigus foliaceus (EPF) in an autoimmune skin disease present in areas of the Amazonia. We report a 36 year-old woman who presented EPF at 17 weeks of pregnancy. At 29 weeks, she started antimicrobial treatment and steroids. At the moment of delivery, the disease was in remission and cutaneous lesions were not seen in the neonate. Indirect immunofluorescence titers of total IgG in the mother and in the neonate were negative. Sixteen months later, IgG titers in the offspring were 1/20 and remained negative in the mother, who was on low doses of oral corticosteroids.


Cardiac resynchronization therapy in patients with chronic atrial fibrillation.
Heart failure (HF) and atrial fibrillation (AF), 2 of the most common cardiovascular disorders, often coexist in the same patient, as 1 condition can lead to the other. The best approach to AF management in patients with HF is currently under investigation, but there seems to be an abundance of evidence in support of cardiac resynchronization therapy (CRT) in this group of patients. In addition, CRT is emerging as a superior option to stand-alone right ventricular pacing in patients with structural heart disease. However, in patients with AF, an adequate rate control is critical for this therapy to be highly effective. As control of the ventricular response can be difficult to achieve in many of those patients, often a nonpharmacologic intervention is required, such as ablation of the atrioventricular node to create heart block. The definitive role for CRT with or without atrioventricular nodal ablation in patients with AF and HF is yet to be studied in large, well-designed, randomized, controlled clinical trials.


Materials in particulate form for tissue engineering. 2. Applications in bone.
Materials in particulate form have been the subjects of intensive research in view of their use as drug delivery systems. While within this application there are still issues to be addressed, these systems are now being regarded as having a great potential for tissue engineering applications. Bone repair is a very demanding task, due to the specific characteristics of skeletal tissues, and the design of scaffolds for bone tissue engineering presents several difficulties. Materials in particulate form are now seen as a means of achieving higher control over parameters such as porosity, pore size, surface area and the mechanical properties of the scaffold. These materials also have the potential to incorporate biologically active molecules for release and to serve as carriers for cells. It is believed that the combination of these features would create a more efficient approach towards regeneration. This review focuses on the application of materials in particulate form for bone tissue engineering. A brief overview of bone biology and the healing process is also provided in order to place the application in its broader context. An original compilation of molecules with a documented role in bone tissue biology is listed, as they have the potential to be used in bone tissue engineering strategies. To sum up this review, examples of works addressing the above aspects are presented.


Towards understanding promiscuity in multidrug efflux pumps.
Drug export from cells is a major factor in the acquisition of cellular resistance to antimicrobial and cancer chemotherapy, and poses a significant threat to future clinical management of disease. Many of the proteins that catalyse drug efflux do so with remarkably low substrate specificity, a phenomenon known as multidrug transport. For these reasons we need a greater understanding of drug recognition and transport in multidrug pumps to inform research that attempts to circumvent their action. Structural and computational studies have been heralded as being great strides towards a full elucidation of multidrug recognition and transport. In this review we summarise these advances and ask how close we are to a molecular understanding of this remarkable phenomenon.