Monocyte disorder causing cellular immunodeficiency: a family study.
We report a familial type of monocyte dysfunction not recognized previously. This disorder was observed in a young adult man with a long clinical history of recurrent, self-limited episodes of cryptogenic fever accompanied by digestive and respiratory symptoms and repeated oral and skin infections. Lectin-induced lymphocyte transformation was reduced and skin tests revealed anergy to tuberculin and candidin. Monocytes from this patient exhibited markedly diminished expression of cytoskeletal vimentin intermediate filaments, HLA-DR antigens and immunological receptors for IgG Fc and C3b. These abnormal monocytes demonstrated impaired phagocytosis and reduced accessory cell function on PHA-mediated lymphocyte activation. Release of soluble lymphocyte-activating factors by these cells was found to be defective. Lymphocytes from the patient responded appropriately to lectin in the presence of normal monocytes. Two family members of the proband presented similar monocyte defects although they only manifested minor clinical symptoms. This syndrome underlines the interest of testing monocyte markers and function in subjects with clinical manifestations of immunodeficiency.


Structure and interdomain interactions of a hybrid domain: a disulphide-rich module of the fibrillin/LTBP superfamily of matrix proteins.
The fibrillins and latent transforming growth factor-beta binding proteins (LTBPs) form a superfamily of structurally-related proteins consisting of calcium-binding epidermal growth factor-like (cbEGF) domains interspersed with 8-cysteine-containing transforming growth factor beta-binding protein-like (TB) and hybrid (hyb) domains. Fibrillins are the major components of the extracellular 10-12 nm diameter microfibrils, which mediate a variety of cell-matrix interactions. Here we present the crystal structure of a fibrillin-1 cbEGF9-hyb2-cbEGF10 fragment, solved to 1.8 A resolution. The hybrid domain fold is similar, but not identical, to the TB domain fold seen in previous fibrillin-1 and LTBP-1 fragments. Pairwise interactions with neighboring cbEGF domains demonstrate extensive interfaces, with the hyb2-cbEGF10 interface dependent on Ca(2+) binding. These observations provide accurate constraints for models of fibrillin organization within the 10-12 nm microfibrils and provide further molecular insights into how Ca(2+) binding influences the intermolecular interactions and biomechanical properties of fibrillin-1.


Involvement of p54(nrb), a PSF partner protein, in DNA double-strand break repair and radioresistance.
Mammalian cells repair DNA double-strand breaks (DSBs) via efficient pathways of direct, nonhomologous DNA end joining (NHEJ) and homologous recombination (HR). Prior work has identified a complex of two polypeptides, PSF and p54(nrb), as a stimulatory factor in a reconstituted in vitro NHEJ system. PSF also stimulates early steps of HR in vitro. PSF and p54(nrb) are RNA recognition motif-containing proteins with well-established functions in RNA processing and transport, and their apparent involvement in DSB repair was unexpected. Here we investigate the requirement for p54(nrb) in DSB repair in vivo. Cells treated with siRNA to attenuate p54(nrb) expression exhibited a delay in DSB repair in a gamma-H2AX focus assay. Stable knockdown cell lines derived by p54(nrb) miRNA transfection showed a significant increase in ionizing radiation-induced chromosomal aberrations. They also showed increased radiosensitivity in a clonogenic survival assay. Together, results indicate that p54(nrb) contributes to rapid and accurate repair of DSBs in vivo in human cells and that the PSF.p54(nrb) complex may thus be a potential target for radiosensitizer development.


Methorexate therapy in a patient with rheumatoid arthritis complicated by idiopathic thrombocytopenic purpura.
The association of rheumatoid arthritis (RA) and immune thrombocytopenic purpura (ITP) has been reported rarely. Methotrexate, which is used for RA treatment, causes thrombocytopenia. Therefore, in medical practice, physicians avoid using methotrexate for RA in patients who have both RA and ITP. Here, we report an RA case that also had ITP, which did not decrease in platelet count after methotrexate therapy. A 50-year-old woman was diagnosed with diabetes mellitus in 1990, RA in 1995, and ITP in 2000. She had received hydroxychloroquine for more than 5 years. She was treated with prednisolone 16 mg/daily between 2006 and 2007, but she discontinued this therapy because of weight gain. Laboratory findings were not remarkable, except for thrombocytopenia. We started methotrexate therapy 10 mg per week for treatment of RA, and hydroxychloroquine therapy was stopped due to nonresponse. The methotrexate dose was increased up to 15 mg/week. Her complete blood cell count was monitored frequently. We did not observe any decrease in platelet count, while active arthritis symptoms of the patient were relieved. This case shows that methotrexate may be used in patients diagnosed with RA that is associated with ITP under strict monitoring.


Suppressing Aedes albopictus, an emerging vector of dengue and chikungunya viruses, by a novel combination of a monomolecular film and an insect-growth regulator.
The Asian tiger mosquito Aedes albopictus (Skuse) is rapidly increasing its global range and importance in transmission of chikungunya and dengue viruses. We tested pellet formulations of a monomolecular film (Agnique) and (S)-methoprene (Altosid) under laboratory and field conditions. In the laboratory, Agnique provided 80% control for 20 days, whereas Altosid, in combination with Agnique, provided 80% control for > 60 days. During field trials, the 1:1 pellet ratio of combined products provided > 95% control for at least 32 days and 50% control for at least 50 days. Altosid remained effective after a 107-day laboratory-induced drought, suggesting that the product serves as a means of control during drought conditions and against spring broods in temperate regions. Agnique and Altosid, when used in tandem for cryptic, difficult-to-treat locations, can provide long-term control of Ae. albopictus larvae and pupae. The possible additive or synergistic effects of the combined products deserve further investigation.


Good laboratory practice: preventing introduction of bias at the bench.
As a research community, we have failed to demonstrate that drugs which show substantial efficacy in animal models of cerebral ischemia can also improve outcome in human stroke. Summary of Review- Accumulating evidence suggests this may be due, at least in part, to problems in the design, conduct and reporting of animal experiments which create a systematic bias resulting in the overstatement of neuroprotective efficacy. Here, we set out a series of measures to reduce bias in the design, conduct and reporting of animal experiments modeling human stroke.


Medication and psychotherapy in eating disorders: is there a gap between research and practice?
Little research has investigated the use of evidence-based guidelines by eating disorder (ED) therapists, or prescribing of psychotropic medication. Moreover, people with EDs have rarely been surveyed on these topics, and their clinical and demographic features have not been presented. This study investigated perception of psychotherapy, psychotropic medication and the clinical characteristics of a community sample of people with EDs. An online survey methodology was used to recruit 253 people with eating disorders in the community. Where feasible, comparisons were made between four types of eating disorder, anorexia nervosa, bulimia nervosa, and two types of atypical or 'sub-threshold' eating disorder. Unlike medication, reported psychotherapy showed some congruence with evidence based and other guidance. Most participants were currently receiving either psychotherapy, medication or both, and most had a severe and chronic ED. Findings are considered in light of use of evidence-based treatment for EDs, calls for greater dissemination of cognitive behaviour therapy (CBT); indications that much may be poor quality; and the importance of what treatments to offer those who are chronically and severely ill. Development of theory and novel treatments is considered a priority.


Prevalence of torture and other warrelated traumatic events in forced migrants: A systematic review.
To describe and appraise the research literature reporting prevalence of torture and/or war-related potentially traumatic experiences (PTEs) in adult forced migrants living in high-income countries. A search for peer-reviewed articles in English was conducted in PubMed, Web of Science, PILOTS, key journals, and reference lists. Studies based on clinical samples and samples where less than half of participants were forced migrants were excluded. Data was extracted and a methodological quality appraisal was performed. A total of 3,470 titles and abstracts were retrieved and screened. Of these, 198 were retrieved in full-text. Forty-one articles fulfilled inclusion criteria and the total number of study participants was 12,020 (median 170). A majority focused on specific ethnic groups or nationalities, Southeast Asian, Middle Eastern and Balkan being the most frequent. Reported prevalence rates of torture ranged between one and 76 % (median 27 %). Almost all participants across all studies had experienced some kind of war-related PTE. Reported prevalence rates of torture and war-related PTEs vary between groups of forced migrants. Trauma history was often studied as a background variable in relation to mental health. The heterogeneity of data, as well as the methodological challenges in reaching forced migrants and defining and measuring traumatic experiences, prevent generalisation concerning trauma history across groups.


Morphologic characterization of syndromic gastric polyps.
The morphology of gastric hamartomatous polyps from patients with juvenile polyposis syndrome (JuvPS) and Peutz-Jeghers' Syndrome (PJS) is poorly characterized. We investigated the histologic features of gastric polyps in patients with established JuvPS or PJS to develop improved histologic criteria to distinguish these from gastric hyperplastic (HP) polyps. The patients with clinically confirmed hamartomatous polyposis syndromes were identified, including 26 patients with JuvPS (both familial and sporadic) and 17 patients with PJS. All gastric polyps (n=30) from these patients were intermixed with gastric HP polyps from nonsyndromic patients (n=26) and subsequently blindly reviewed by a panel of gastrointestinal pathologists. A consensus diagnosis was rendered. The panel then reviewed the slides in the context of clinical data and identified histologic features for distinguishing JuvPS, PJS, and HP gastric polyps based on epithelial changes, pit architecture, lamina propria features, and smooth muscle qualities. A sleeping period of 6 months lapsed before the same cases were renumbered and blindly rereviewed independently. Diagnoses were then rendered while adhering to the suggested criteria. Cases that the reviewers recalled were discarded from the study (n=8). On initial review, accuracy in diagnosis of gastric polyps in JuvPS was 50% and was 18% in PJS compared with 92% for HP gastric polyps. Adherence to the recommended histologic criteria resulted in diagnostic accuracy of 41% for JuvPS and 54% for PJS, compared with 73% for HP gastric polyps. Accuracy in diagnosis in antral mucosa was 66%, oxyntic mucosa 71%, and transitional-type mucosa (mixed antral and oxyntic) 32%. The diagnostic accuracy based on polyp size was 59% for polyps which were less than equal to 3 mm, 56% for those 4 to 9 mm, and 81% for polyps which were more than equal to 10 mm. The identification of gastric polyps from JuvPS and PJS patients without the context of clinical history of these syndromes remains poor, even with adherence to a set of morphologic criteria. Abiding by such criteria improved recognition of PJS polyps by more than double (P<0.19), but yielded an accuracy of only 54%. The accuracy did not improve when results were stratified for polyp location but did with biopsy size which were more than equal to10 mm. Whereas these syndromic polyps are readily diagnosed in the small bowel and colon, histologic features to distinguish gastric JuvPS and PJS from gastric HP polyps are unreliable.


Premolar bracket position revised: proximal and occlusal contacts assessment.
To assess the effect of conventional bracket slot vertical position (4 mm from incisal edge) on crown spatial orientation relative to adjacent and opposing teeth through proximal contact balance and occlusal contacts assessment. Forty patients were divided into two groups; in the first group, brackets were bonded 4 mm from the intact incisal edge; in the second group, bracket position was determined individually by transferring the maximum embrasure difference to the bracket slot height difference. The difference of proximal contacts between premolars and molars was measured. Bite registration with the use of articulating paper was also performed on all patients. Traces indicating contact between premolar maxillary lingual cusps and opposing mandibular fossa of the first and second premolars were evaluated. Differences with respect to the distribution of unbalanced proximal contacts and negative contacts between the two bonding schemes were analyzed with the chi-square test at P = .05 level of significance. Aligning the bracket slot with the center of the clinical crown (FA point) consistently produced unbalanced proximal contacts in excess of 1 mm, as well as increased absence of occlusal contacts. Positioning the bracket slot on the center of the premolar crown may result in marginal ridge discrepancy between the premolars and molars and lack of occlusal contacts with opposing dentition. This may be due to the height difference noted between the buccal and lingual cusp of the premolars, as well as variation in molar tube or band placement. Functional occlusal analysis should be integrated in the finishing stages of treatment to facilitate proper premolar crown position.


Rhodamine spiroamides for multicolor single-molecule switching fluorescent nanoscopy.
The design, synthesis, and evaluation of new rhodamine spiroamides are described. These molecules have applications in optical nanoscopy based on random switching of the fluorescent single molecules. The new markers may be used in (co)localization studies of various objects and their (mutual) positions and shape can be determined with a precision of a few tens of nanometers. Multicolor staining, good photoactivation, a large number of emitted photons, and selective chemical binding with amino or thiol groups were achieved due to the presence of various functional groups on the rhodamine spiroamides. Rigidized sulfonated xanthene fragment fused with six-membered rings, N,N'-bis(2,2,2-trifluoroethyl) groups, and a combination of additional double bonds and sulfonic acid groups with simple aliphatic spiroamide residue provide multicolor properties and improve performance of the rhodamine spiroamides in photoactivation and bioconjugation reactions. Having both essential parts of the photoswitchable assembly-the switching and the fluorescent (reporter) groups-combined in one chemical entity make this approach attractive for further development. A series of rhodamine spiroamides is presented along with characterizations of their most relevant properties for application as fluorescent probes in single-molecule switching and localization microscopy. Optical images with resolutions on the nanometer scale illustrate the potential of the labels in the colocalization of biological objects and the two-photon activation technique with optical sectioning.


A full systematic review was completed in 2 weeks using automation tools: a case study.
Systematic reviews (SRs) are time and resource intensive, requiring approximately 1 year from protocol registration to submission for publication. Our aim was to describe the process, facilitators, and barriers to completing the first 2-week full SR. We systematically reviewed evidence of the impact of increased fluid intake, on urinary tract infection (UTI) recurrence, in individuals at risk for UTIs. The review was conducted by experienced systematic reviewers with complementary skills (two researcher clinicians, an information specialist, and an epidemiologist), using Systematic Review Automation tools, and blocked off time for the duration of the project. The outcomes were time to complete the SR, time to complete individual SR tasks, facilitators and barriers to progress, and peer reviewer feedback on the SR manuscript. Times to completion were analyzed quantitatively (minutes and calendar days); facilitators and barriers were mapped onto the Theoretical Domains Framework; and peer reviewer feedback was analyzed quantitatively and narratively. The SR was completed in 61 person-hours (9 workdays; 12 calendar days); accepted version of the manuscript required 71 person-hours. Individual SR tasks ranged from 16 person-minutes (deduplication of search results) to 461 person-minutes (data extraction). The least time-consuming SR tasks were obtaining full-texts, searches, citation analysis, data synthesis, and deduplication. The most time-consuming tasks were data extraction, write-up, abstract screening, full-text screening, and risk of bias. Facilitators and barriers mapped onto the following domains: knowledge; skills; memory, attention, and decision process; environmental context and resources; and technology and infrastructure. Two sets of peer reviewer feedback were received on the manuscript: the first included 34 comments requesting changes, 17 changes were made, requiring 173 person-minutes; the second requested 13 changes, and eight were made, requiring 121 person-minutes. A small and experienced systematic reviewer team using Systematic Review Automation tools who have protected time to focus solely on the SR can complete a moderately sized SR in 2 weeks.


I-cell disease (mucolipidosis II): resolution of N-acetyl-beta-D-glucosaminidase and alpha-L-fucosidase components by DEAE-cellulose chromatography.
1. In a patient with I-cell disease the activities of several acid hydrolases were elevated inplasma and reduced in cultured fibroblasts when compared with normal values. Normal activities for the enzymes were found in leucocytes. These findings agree with reports on other cases. 2. N-Acetyl-beta-D-glucosaminidase was resolved into its component forms by chromatography on microcolumns of DEAE-cellulose coupled with continuous automated assay of activity in the column effluent. Cultured skin fibroblasts from three patients showed a profound deficiency of glucosaminidase component A and a relative increase in the activity of a form eluted earlier than A. 3. In the one patient studied, the elution profile of plasma glucosaminidase was similar to that of normal plasma, but treatment with neuraminidase revealed a minor component which did not appear in control specimens. 4. Chromatographic resolution of glucosaminidase secreted by normal fibroblasts into the culture medium shoed that component A comprised two forms, a serum-type and a tissue-type, whereas only a serum-type was found in I-cell medium. 5. Different forma of alpha-L-fucosidase were shown to occur in normal plasma and fibroblasts. This is the second lysosomal hydrolase for which differences between intracellular and extracellular forms have been described and might reflect a general phenomenon. 6. The major acidic component of fucosidase from normal fibroblasts was not detected in I-cell fibroblasts. Elution profiles of fucosidase activity in normal and I-cell plasma were indistinguishable, both before and after treatment with neuraminidase. 7. On the basis of the above findings, we suggest that for several acid hydrolases there is a common biosynthetic reaction, which produces forms of these enzymes destined for incorporation into primary lysosomes rather than secretion by the cell. In cultured fibroblasts from patients with I-cell disease, the enzyme catalysing the reaction leading to the production of intracellular forms is deficient or defective, whereas the synthesis of precursor and secreted forms is unaffected.


[Personality traits in adults with autism spectrum disorders measured by means of the Temperament and Character Inventory].
There is growing interest in autism spectrum disorders (asd) in adulthood. Someone can be diagnosed with ASD, but the diagnosis tells us very little about the patient's temperament, character and personality. Comparatively little is known about the personality traits of persons with ASD. To map personality traits of persons with asd. The Temperament and Character Inventory (tci) was administered to a group of 68 men diagnosed with asd at the Lucertis Sarr expertise centre for Autism and at the Department of Psychiatry, Erasmus mc, Rotterdam, the Netherlands. The control group, specified in the instructions for the tci, consisted of a group of 447 men from the general population. Compared to the control group, men with asd scored higher on the scale Harm Avoidance, but lower on Sociability, Novelty Seeking, Reward Dependence, Self-directedness, and Cooperativeness. The score pattern found in men with asd is consistent with the clinical picture of asd and corresponds to earlier results of research done in Sweden. In our study we argue that negatively interpreted temperament and character traits can often be interpreted in a positive way.


METAREP: JCVI metagenomics reports--an open source tool for high-performance comparative metagenomics.
JCVI Metagenomics Reports (METAREP) is a Web 2.0 application designed to help scientists analyze and compare annotated metagenomics datasets. It utilizes Solr/Lucene, a high-performance scalable search engine, to quickly query large data collections. Furthermore, users can use its SQL-like query syntax to filter and refine datasets. METAREP provides graphical summaries for top taxonomic and functional classifications as well as a GO, NCBI Taxonomy and KEGG Pathway Browser. Users can compare absolute and relative counts of multiple datasets at various functional and taxonomic levels. Advanced comparative features comprise statistical tests as well as multidimensional scaling, heatmap and hierarchical clustering plots. Summaries can be exported as tab-delimited files, publication quality plots in PDF format. A data management layer allows collaborative data analysis and result sharing. Web site http://www.jcvi.org/metarep; source code http://github.com/jcvi/METAREP CONTACT: email@example.com Supplementary data are available at Bioinformatics online.


Taxonomic and phylogenetic status of non-tuberculous mycobacteria in a Caribbean setting.
This report describes detailed taxonomic and phylogenetic analysis of 15 non-tuberculous mycobacteria (NTMs) isolated from human pathological specimens in a Caribbean setting (12 slow-growers and three rapid-growers) that were not identified by cultural and biochemical tests and drug-susceptibility results. These isolates were further studied using PCR restriction fragment length polymorphism analysis (PRA) of a 441bp hsp65 fragment, as well as the sequencing of 16S rDNA and hsp65 DNA, and HPLC of the mycolic acids. Our results showed that taxonomic position of well-defined NTMs was resolved by PRA and sequencing of hsp65, nonetheless, it was not suitable to investigate rarely observed or new strains that required 16S rDNA sequencing and HPLC for a definite response. Unrooted neighbor-joining phylogenetic trees were drawn based upon the 16S rDNA and hsp65 sequences of the 15 NTMs compared with those from described species (73 for 16S rDNA and 45 for hsp65). For most of the NTMs not showing an exactly matching sequence with either hsp65 or 16S rDNA in the GenBank, the phylogenetic tree was able to provide with useful indications about their relatedness to known species. In such a case, a concording HPLC pattern with the sequence data and the place of the strain within the tree could lead to a potential identification. We also identified three identical isolates that define a new mycobacterial species within the group of M. simiae-related mycobacteria. The isolation and characterization of mycobacteria from new settings may lead to identify potential pathogens that may propogate in future because of increased human migration, travels, and climatic and ecological changes of the modern world.


High pollutant removal efficacy of a large constructed wetland leads to receiving stream improvements.
Hewletts Creek, in Wilmington, North Carolina, drains a large suburban watershed and as such is affected by high fecal bacteria loads and periodic algal blooms from nutrient loading. During 2007, a 3.1-ha wetland was constructed to treat stormwater runoff from a 238-ha watershed within the Hewletts Creek drainage. A rain event sampling program was performed in 2009-2010 to evaluate the efficacy of the wetland in reducing pollutant loads from the stormwater runoff passing through the wetland. During the eight storms sampled, the wetland greatly moderated the hydrograph and retained and/or removed 50 to 75% of the inflowing stormwater volume. High removal rates of fecal coliform bacteria were achieved, with an average load reduction of 99% and overall concentration reduction of >90%. Particularly high (>90%) reductions of ammonium and orthophosphate loads also occurred, and lesser but still substantial reductions of total phosphorus (89%) and total suspended solids loads (88%) were achieved. Removal of nitrate was seasonally dependent, with lower removal occurring in cold weather and a high percentage (90%+) of nitrate load removal occurring in the growing season when water temperature exceeded 15°C. Long-term before-and-after sampling in downstream Hewletts Creek proper showed that, after wetland construction, statistically significant average decreases of 43% for nitrate, 72% for ammonium, and 59% for fecal coliform bacteria were realized. Wetland features contributing to the high pollutant control efficacy included available space for a large wetland, construction of deep forebays, and a dense and diverse aquatic and shoreline plant assemblage.


Effect of selected bisphenol derivatives on nuclear receptor expression in ovarian cell line COV434.
Objectives. Bisphenol A (BPA), as an indispensable plastic additive, has also been proven as an endocrine disruptor associated with adverse health effects including impaired ovarian function and cancer. Due to the restrictions of its usage, several analogs have been employed to replace BPA. Although many studies revealed a harmfulness in the biological effects of BPA analogs, their specific targets remain largely unknown. Nuclear receptors (NRs) may be one of the most important targets of bisphenols. Therefore, in this study, our attention was directed to explore the effect of BPA and its analogs, AF and S, on the mRNA expression of selected NRs involved in the steroidogenic and carcinogenic pathways in the human granulosa cell line COV434. The NRs investigated included: thyroid hormone receptor α (THRA), peroxisome proliferator activating receptor β/δ (PPARD), retinoid X receptor α (RXRA), chicken ovalbumin upstream promoter-transcription factor II (COUPTFII), nuclear receptor-related protein 1 (NURR1), and liver receptor homolog-1 (LRH1).Methods. COV434 cells were treated with the bisphenols at the concentrations of 10-9 M, 10-7 M, and 10-5 M, and after 24 and 48 h, cell viability was monitored by the MTS assay and gene expressions were analyzed using RT-qPCR.Results. Bisphenol treatment did not alter the COV434 cell viability. After 24 h, the expression of neither of the NRs was changed. Likewise, after 48 h, the expression of the selected genes was not altered. However, both BPAF and BPS increased, at the highest concentration (10-5 M) used, the mRNA levels of both PPARD and NURR1 NRs after 48 h of the treatment. In the BPA-treated groups, no significant upregulation was observed.Conclusions. In the present study, the effect of bisphenols on COUP-TFII, Nurr1, and LRH-1 NRs was investigated for the first time. Although generally we did not observe that BPs provoked any alterations in the expression of the selected NRs in COV434 cells, at specific concentrations and time points they might alter mRNA expression of certain NRs (NURR1, PPARD).


Axonal loss and gray matter pathology as a direct result of autoimmunity to neurofilaments.
Axonal damage is considered the major cause of irreversible disability in multiple sclerosis (MS). Which mechanisms underlie the damage and whether this is secondary to myelin damage remains to be clarified. Recently, we have demonstrated that autoimmunity to the axonal/neuronal cytoskeletal protein neurofilament light (NF-L) induces axonal damage and neurological disease including spasticity - a common feature of MS. To examine the relationship between axonal damage and demyelination we have characterized the detailed neuropathology of NF-L-induced disease in Biozzi mice compared to classical experimental autoimmune encephalomyelitis (EAE) induced with myelin oligodendrocyte glycoprotein (MOG). In NF-L-induced neurological disease the lesions were predominantly located in the dorsal column displaying extensive axonal degeneration, but were also abundant in the gray matter. In contrast, lesions in MOG-EAE were restricted to the lateral and ventral columns and displayed less axonal damage and little gray matter involvement. The differential lesion location was confirmed by quantitation of leukocyte subsets. In both diseases myelin damage was a common feature although the numerous empty myelin sheaths in NF-L-disease indicative of axonal damage suggest that myelin damage was a secondary event. In summary, autoimmunity to NF-L induces a distinct lesion topology, axonal damage and gray matter lesions supporting the notion that axonal loss and gray matter pathology can be the direct consequence of a primary autoimmune attack against axonal antigens such as NF-L rather than merely a secondary event to myelin damage.


Human mitochondrial DNA polymerase holoenzyme: reconstitution and characterization.
We have reconstituted the holoenzyme of the human mitochondrial DNA polymerase from cloned and overexpressed catalytic and accessory subunits. We have examined the polymerization activity of the catalytic subunit alone and of the holoenzyme to establish the function of the accessory subunit in this two subunit enzyme. The accessory subunit associates with the catalytic subunit with a dissociation constant of 35 +/- 16 nM as measured by the concentration dependence of its effect in stimulating maximal DNA binding and polymerization. At saturating concentrations, the accessory subunit contributes to every kinetic parameter examined to facilitate tighter binding of DNA and nucleotide and faster replication. The accessory protein makes the DNA binding 3.5-fold tighter (K(d) of 9.9 +/- 2.1 nM compared to 39 +/- 10 nM for the catalytic subunit alone) without significantly affecting the DNA dissociation rate (0.02 +/- 0.001 compared to 0.03 +/- 0.001 s(-)(1)). The ground-state nucleotide binding is improved from 4.7 +/- 2.0 to 0.78 +/- 0.065 microM, and the maximum DNA polymerization rate is increased from 8.7 +/- 1.1 to 45 +/- 1 s(-)(1) by the addition of the accessory protein. This leads to an increase in processivity from an estimated 290 +/- 46 to 2250 +/- 162. Although the accessory protein has been described as a "processivity factor" because of its effect on the ratio of rate constants defining processivity, this terminology falls short of adequately describing the profound effects of the small subunit on nucleotide-binding and incorporation catalyzed by the large subunit. By using the complete holoenzyme, we can now proceed with a comprehensive analysis of the structural and mechanistic determinants of enzyme specificity that govern toxicity of nucleoside analogues used in the treatment of viral infections such as AIDS.


Probing Primordial Black Hole Dark Matter with Gravitational Waves.
Primordial black holes (PBHs) have long been suggested as a candidate for making up some or all of the dark matter in the Universe. Most of the theoretically possible mass range for PBH dark matter has been ruled out with various null observations of expected signatures of their interaction with standard astrophysical objects. However, current constraints are significantly less robust in the 20 M_{⊙}≲M_{PBH}≲100 M_{⊙} mass window, which has received much attention recently, following the detection of merging black holes with estimated masses of ∼30 M_{⊙} by LIGO and the suggestion that these could be black holes formed in the early Universe. We consider the potential of advanced LIGO (aLIGO) operating at design sensitivity to probe this mass range by looking for peaks in the mass spectrum of detected events. To quantify the background, which is due to black holes that are formed from dying stars, we model the shape of the stellar-black-hole mass function and calibrate its amplitude to match the O1 results. Adopting very conservative assumptions about the PBH and stellar-black-hole merger rates, we show that ∼5 yr of aLIGO data can be used to detect a contribution of >20 M_{⊙} PBHs to dark matter down to f_{PBH}<0.5 at >99.9% confidence level. Combined with other probes that already suggest tension with f_{PBH}=1, the obtainable independent limits from aLIGO will thus enable a firm test of the scenario that PBHs make up all of dark matter.


Inhibition of membrane Na+-K+ ATPase activity: a common pathway in central nervous system disorders.
The study was conducted to assess the role of hypothalamic digoxin in neuropsychiatric and systemic disorders. A hypothesis regarding the central role of hypothalamic digoxin in neuroimmunoendocrine integration is proposed. Blood samples from patients of CNS glioma, multiple sclerosis, systemic lupus erythematosis, subacute sclerosing panencephalitis, primary generalized epilepsy, Parkinson's disease, Down syndrome, AIDS dementia with neuropsychiatric features, syndrome X with multiple lacunar state, senile dementia, familial group (a family with familial coexistence of schizophrenia, Parkinson's disease, primary generalized epilepsy, malignant neoplasia, rheumatoid arthritis and syndrome X over three generations), schizophrenia and manic depressive psychosis were analysed for RBC membrane Na+-K+ ATPase, levels of digoxin and Mg++. Inhibition of RBC membrane Na+-K+ ATPase activity was observed in most cases along with increase in the levels of serum digoxin and decrease in the level of serum Mg++. The decreased Na+-K+ ATPase activity can be due to increased digoxin, which is a potent inhibitor of this enzyme. The inhibition of Na+-K+ ATPase can contribute to increase in intracellular calcium and decrease in magnesium, which can result in 1) defective neurotransmitter transport mechanism, 2) neuronal degeneration and apoptosis, 3) mitochondrial dysfunction, 4) defective golgi body function and protein processing dysfunction, 5) immune dysfunction and oncogenesis. The mechanism of how increased intracellular calcium and decreased magnesium can contribute to the above effects is discussed.


Irrigation and enhanced soil carbon input effects on below-ground carbon cycling in semiarid temperate grasslands.
Global climate change is generally expected to increase net primary production, resulting in increased soil carbon (C) inputs. To gain an understanding of how such increased soil C inputs would affect C cycling in the vast grasslands of northern China, we conducted a field experiment in which the responses of plant and microbial biomass and respiration were studied. Our experiment included the below-ground addition of particulate organic matter (POM) at rates equivalent to 0, 60, 120 and 240 g C m(-2), under either natural precipitation or under enhanced precipitation during the summer period (as predicted for that region in recent simulations using general circulation models). We observed that addition of POM had a large effect on soil microbial biomass and activity and that a major part of the added C was rapidly lost from the system. This suggests that microbial activity in the vast temperate grassland ecosystems of northern China is energy-limited. Moreover, POM addition (and the associated nutrient release) affected plant growth much more than the additional water input. Although we performed no direct fertilization experiments, the response of plant productivity to POM addition (and associated release of nutrients) leads us to believe that plant productivity in the semiarid grassland ecosystems of northern China is primarily limited by nutrients and not by water.


Clinicians' expectations of Web 2.0 as a mechanism for knowledge transfer of stroke best practices.
Health professionals are increasingly encouraged to adopt an evidence-based practice to ensure greater efficiency of their services. To promote this practice, several strategies exist: distribution of educational materials, local consensus processes, educational outreach visits, local opinion leaders, and reminders. Despite these strategies, gaps continue to be observed between practice and scientific evidence. Therefore, it is important to implement innovative knowledge transfer strategies that will change health professionals' practices. Through its interactive capacities, Web 2.0 applications are worth exploring. As an example, virtual communities of practice have already begun to influence professional practice. This study was initially developed to help design a Web 2.0 platform for health professionals working with stroke patients. The aim was to gain a better understanding of professionals' perceptions of Web 2.0 before the development of the platform. A qualitative study following a phenomenological approach was chosen. We conducted individual semi-structured interviews with clinicians and managers. Interview transcripts were subjected to a content analysis. Twenty-four female clinicians and managers in Quebec, Canada, aged 28-66 participated. Most participants identified knowledge transfer as the most useful outcome of a Web 2.0 platform. Respondents also expressed their need for a user-friendly platform. Accessibility to a computer and the Internet, features of the Web 2.0 platform, user support, technology skills, and previous technological experience were found to influence perceived ease of use and usefulness. Our results show that the perceived lack of time of health professionals has an influence on perceived behavioral intention to use it despite favorable perception of the usefulness of the Web 2.0 platform. In conclusion, female health professionals in Quebec believe that Web 2.0 may be a useful mechanism for knowledge transfer. However, lack of time and lack of technological skills may limit their use of a future Web 2.0 platform. Further studies are required with other populations and in other regions to confirm these findings.


The Scientific Foundation for personal genomics: recommendations from a National Institutes of Health-Centers for Disease Control and Prevention multidisciplinary workshop.
The increasing availability of personal genomic tests has led to discussions about the validity and utility of such tests and the balance of benefits and harms. A multidisciplinary workshop was convened by the National Institutes of Health and the Centers for Disease Control and Prevention to review the scientific foundation for using personal genomics in risk assessment and disease prevention and to develop recommendations for targeted research. The clinical validity and utility of personal genomics is a moving target with rapidly developing discoveries but little translation research to close the gap between discoveries and health impact. Workshop participants made recommendations in five domains: (1) developing and applying scientific standards for assessing personal genomic tests; (2) developing and applying a multidisciplinary research agenda, including observational studies and clinical trials to fill knowledge gaps in clinical validity and utility; (3) enhancing credible knowledge synthesis and information dissemination to clinicians and consumers; (4) linking scientific findings to evidence-based recommendations for use of personal genomics; and (5) assessing how the concept of personal utility can affect health benefits, costs, and risks by developing appropriate metrics for evaluation. To fulfill the promise of personal genomics, a rigorous multidisciplinary research agenda is needed.


Epidemiology of diabetic retinopathy, diabetic macular edema and related vision loss.
Diabetic retinopathy (DR) is a leading cause of vision-loss globally. Of an estimated 285 million people with diabetes mellitus worldwide, approximately one third have signs of DR and of these, a further one third of DR is vision-threatening DR, including diabetic macular edema (DME). The identification of established modifiable risk factors for DR such as hyperglycemia and hypertension has provided the basis for risk factor control in preventing onset and progression of DR. Additional research investigating novel risk factors has improved our understanding of multiple biological pathways involved in the pathogenesis of DR and DME, especially those involved in inflammation and oxidative stress. Variations in DR prevalence between populations have also sparked interest in genetic studies to identify loci associated with disease susceptibility. In this review, major trends in the prevalence, incidence, progression and regression of DR and DME are explored, and gaps in literature identified. Established and novel risk factors are also extensively reviewed with a focus on landmark studies and updates from the recent literature.


Characteristics of compacted plasmid DNA by high pressurization.
In order to investigate the effect of pressure on the tertiary structure of plasmid DNA having the supercoiled and relaxed forms, the solution of plasmid DNA was hydrostatically pressurized at different atmosphere and 40 degrees C for various times. For dynamic light scattering (DLS) measurement of the pressurized plasmid DNA, the hydrodynamic diameters of the super-coiled and relaxed plasmid DNA were decreased with increasing pressure. Also, at constant pressure, a long period of pressure treatment effectively induced the decrease in plasmid DNA. These results suggest that the plasmid DNA was condensed by high hydrostatic pressurization. The circular dichroism (CD) spectrum of the pressurized plasmid DNA was slightly changed. For digestion by S1 nuclease, which selectively cleaves single strand DNA, the pressurized plasmid DNA was easily degraded compared to the non-pressurized plasmid DNA, suggesting that the double helix of plasmid DNA was partly dissociated to single strand by the pressure-induced compaction of plasmid DNA. These results indicate that high hydrostatic pressurization is one of powerful tools for preparing the compacted plasmid DNA.


The effect of a combination of medetomidine-butorphanol and medetomidine, butorphanol, atropine on glomerular filtration rate in dogs.
The effects of intramuscularly administered medetomidine and butorphanol (MB), and medetomidine, butorphanol, atropine (MBA) on glomerular filtration rate (GFR) were determined in six dogs as measured by 99m-Tc-labeled diethylenetriaminepentaacetic acid (99mTc-DTPA) nuclear scintigraphy. Direct systolic, diastolic, and mean arterial blood pressures and heart rate were measured at regular time intervals before, during, and after GFR calculations. The mean GFR measurement following MB was significantly greater (4.44 ml/min/kg) than following MBA (3.82 ml/min/kg) or saline treatment (3.41 ml/min/kg). There was no significant difference between the mean GFR measurements following MBA injection and following saline injection. Diastolic and mean arterial pressures following MBA injection were significantly higher than the values recorded after either MB or saline alone. Heart rate following MB administration was significantly lower than that recorded for dogs receiving MBA or saline alone. The results of this study indicate that the administration of medetomidine in combination with butorphanol significantly increases total GFR in healthy dogs, while the administration of the combination of medetomidine, butorphanol, and atropine does not.


Antiviral and Immunomodulatory Activity of Silver Nanoparticles in Experimental RSV Infection.
Respiratory syncytial virus (RSV) is an important etiological agent of respiratory infection in children for which no specific treatment option is available. The RSV virion contains two surface glycoproteins (F and G) that are vital for the initial phases of infection, making them critical targets for RSV therapeutics. Recent studies have identified the broad-spectrum antiviral properties of silver nanoparticles (AgNPs) against respiratory pathogens, such as adenovirus, parainfluenza, and influenza. AgNPs achieve this by attaching to viral glycoproteins, blocking entry into the host cell. The objective of this study was to evaluate the antiviral and immunomodulatory effects of AgNPs in RSV infection. Herein we demonstrate AgNP-mediated reduction in RSV replication, both in epithelial cell lines and in experimentally infected BALB/c mice. Marked reduction in pro-inflammatory cytokines (i.e., IL-1α, IL-6, TNF-α) and pro-inflammatory chemokines (i.e., CCL2, CCL3, CCL5) was also observed. Conversely, CXCL1, G-CSF, and GM-CSF were increased in RSV-infected mice treated with AgNPs, consistent with an increase of neutrophil recruitment and activation in the lung tissue. Following experimental antibody-dependent depletion of neutrophils, the antiviral effect of AgNPs in mice treated was ablated. To our knowledge, this is the first in vivo report demonstrating antiviral activity of AgNPs during RSV infection.


Estimating the population attributable fraction for schizophrenia when Toxoplasma gondii is assumed absent in human populations.
There is increasing evidence that infection with Toxoplasma gondii, a common parasite of people, cats and rodents, is associated with an increased risk of a diagnosis of schizophrenia. Although the claim that infection with T. gondii is one of the component causes of a diagnosis of schizophrenia remains contentious it is worth asking how important a causal association might be if only to inform our attitude to further work on the subject. The appropriate measure of importance is the population attributable fraction (PAF). The PAF is the proportion of diagnoses of schizophrenia that would not occur in a population if T. gondii infections were not present. The assumptions that underlie the derivation of the standard formula for measuring the PAF are violated in the specific instance of T. gondii and schizophrenia and so the conventional estimation method cannot be used. Instead, the PAF was estimated using a deterministic model of Toxoplasma gondii infection and schizophrenia occurrence in a hypothetical cohort of people at risk of both conditions. The incidence of infection with T. gondii in the cohort was assumed to be constant. Under these circumstances, the life-time mean population attributable fraction was estimated to be 21.4%, but it could not be ruled out that it could be as high as 30.6% or as low as 13.7% given the 95% confidence interval pertaining to the point estimate of the OR that was central to the calculation. These estimates (even the lowest) are higher than those obtained using the standard method for the same system and underscore the importance of understanding the limitations of conventional epidemiological formulae.


Angioblast-mesenchyme induction of early kidney development is mediated by Wt1 and Vegfa.
Most studies on kidney development have considered the interaction of the metanephric mesenchyme and the ureteric bud to be the major inductive event that maintains tubular differentiation and branching morphogenesis. The mesenchyme produces Gdnf, which stimulates branching, and the ureteric bud stimulates continued growth of the mesenchyme and differentiation of nephrons from the induced mesenchyme. Null mutation of the Wt1 gene eliminates outgrowth of the ureteric bud, but Gdnf has been identified as a target of Pax2, but not of Wt1. Using a novel system for microinjecting and electroporating plasmid expression constructs into murine organ cultures, it has been demonstrated that Vegfa expression in the mesenchyme is regulated by Wt1. Previous studies had identified a population of Flk1-expressing cells in the periphery of the induced mesenchyme, and adjacent to the stalk of the ureteric bud, and that Vegfa was able to stimulate growth of kidneys in organ culture. Here it is demonstrated that signaling through Flk1 is required to maintain expression of Pax2 in the mesenchyme of the early kidney, and for Pax2 to stimulate expression of Gdnf. However, once Gdnf stimulates branching of the ureteric bud, the Flk1-dependent angioblast signal is no longer required to maintain branching morphogenesis and induction of nephrons. Thus, this work demonstrates the presence of a second set of inductive events, involving the mesenchymal and angioblast populations, whereby Wt1-stimulated expression of Vegfa elicits an as-yet-unidentified signal from the angioblasts, which is required to stimulate the expression of Pax2 and Gdnf, which in turn elicits an inductive signal from the ureteric bud.


Atrial Fibrillation Complicating Congestive Heart Failure: Electrophysiological Aspects And Its Deleterious Effect On Cardiac Resynchronization Therapy.
More successful recognition and treatment of cardiovascular risk factors and diseases continues to decrease mortality and increase the proportion of elderly population. Therefore, there are more people with increased risk of developing heart failure and atrial fibrillation in the course of their lives. Atrial fibrillation (AF) can complicate the course of congestive heart failure (HF) leading to acute pulmonary edema. The prevalence of AF, in patients with heart failure, increases with the severity of the disease, reaching up to 40% in advanced cases. In these HF patients, AF is an independent predictor of morbidity and mortality increasing the risk of death and hospitalization. Despite the excellent results obtained with different drugs, the optimal medical treatment can fail in the intention to improve symptoms and quality of life of patients with severe HF. Thus, the necessity to use cardiac devices emerges facing the failure of optimal medical treatment in order to achieve hemodynamic improvement and correction of the physiopathological alterations. Cardiac resynchronization therapy (CRT) can reduce the interventricular and intraventricular mechanical dissynchrony in HF patients. It has been shown that CRT increases the left ventricular filling time, decreases septal dissynchrony, mitral regurgitation, and left ventricular volumes allowing a hemodynamic improvement. However, the development of AF in this setting can avoid the beneficial effects of CRT. Therefore, this manuscript will review the available data on this topic, the electrophysiological aspects of AF, to determine what can be done in the event of an AF complicating congestive HF in CRT patients.


Cordyceps militaris Fruit Body Extract Decreases Testosterone Catabolism and Testosterone-Stimulated Prostate Hypertrophy.
The androgens testosterone and dihydrotestosterone (DHT) are essential for a variety of systemic functions in mature males. Alteration of these hormones results in late-onset hypogonadism (LOH) and benign prostate hyperplasia (BPH). The fruit bodies of fungi of the genus Cordyceps have been regarded as folk medicine or health food with tonic and antifatigue effects. The extract from the fruit body of Cordyceps militaris parasitizing Samia cynthia ricini (CM) was evaluated as a novel-candidate natural product for ameliorating male andropause symptoms. To explore the effects of CM on LOH and BPH, CM was applied to rat models and cultured testicular cells and prostate cells. The concentrations of androgens in the serum and culture media were determined by ELISA. Expression of steroidogenic enzymes and androgen-related genes was evaluated by qPCR, and prostatic cell proliferation was assessed with the cell-viability assay. CM maintained the serum levels of testosterone and DHT, but inhibited testosterone-induced prostate hypertrophy. CM also increased the secretion of testosterone and DHT by primary testicular cells, with no changes in the mRNA expression of steroidogenic enzymes, but decreased the growth of prostatic cell lines. Our data suggest that CM could improve both LOH and BPH in males.


Biological, physiological, pathophysiological, and pharmacological aspects of ghrelin.
Ghrelin is a peptide predominantly produced by the stomach. Ghrelin displays strong GH-releasing activity. This activity is mediated by the activation of the so-called GH secretagogue receptor type 1a. This receptor had been shown to be specific for a family of synthetic, peptidyl and nonpeptidyl GH secretagogues. Apart from a potent GH-releasing action, ghrelin has other activities including stimulation of lactotroph and corticotroph function, influence on the pituitary gonadal axis, stimulation of appetite, control of energy balance, influence on sleep and behavior, control of gastric motility and acid secretion, and influence on pancreatic exocrine and endocrine function as well as on glucose metabolism. Cardiovascular actions and modulation of proliferation of neoplastic cells, as well as of the immune system, are other actions of ghrelin. Therefore, we consider ghrelin a gastrointestinal peptide contributing to the regulation of diverse functions of the gut-brain axis. So, there is indeed a possibility that ghrelin analogs, acting as either agonists or antagonists, might have clinical impact.


Streptococcal M protein: a multipotent and powerful inducer of inflammation.
Severe infections with Streptococcus pyogenes, an important human pathogen, are associated with massive inflammatory reactions in the human host. Here we show that streptococcal M protein interacts with TLR2 on human peripheral blood monocytes. As a consequence, monocytes express the cytokines IL-6, IL-1beta, and TNF-alpha. This response is significantly increased in the presence of neutrophil-derived heparin-binding protein (HBP), which co-stimulates monocytes by interacting with CD11/CD18. Analysis of tissue biopsies from patients with necrotizing fasciitis revealed recruitment of neutrophils and monocytes to the infectious site, combined with the release of HBP. The results show that M protein, in synergy with HBP, evokes an inflammatory response that may contribute to the profound pathophysiological consequences seen in severe streptococcal infections.


The absence of tertiary interactions in a self-assembled DNA crystal structure.
DNA is a highly effective molecule for controlling nanometer-scale structure. The convenience of using DNA lies in the programmability of Watson-Crick base-paired secondary interactions, useful both to design branched molecular motifs and to connect them through sticky-ended cohesion. Recently, the tensegrity triangle motif has been used to self-assemble three-dimensional crystals whose structures have been determined; sticky ends were reported to be the only intermolecular cohesive elements in those crystals. A recent communication in this journal suggested that tertiary interactions between phosphates and cytosine N(4) groups are responsible for intermolecular cohesion in these crystals, in addition to the secondary and covalent interactions programmed into the motif. To resolve this issue, we report experiments challenging this contention. Gel electrophoresis demonstrates that the tensegrity triangle exists in conditions where cytosine-PO(4) tertiary interactions seem ineffective. Furthermore, we have crystallized a tensegrity triangle using a junction lacking the cytosine suggested for involvement in tertiary interactions. The unit cell is isomorphous with that of a tensegrity triangle crystal reported earlier. This structure has been solved by molecular replacement and refined. The data presented here leave no doubt that the tensegrity triangle crystal structures reported earlier depend only on base pairing and covalent interactions for their formation.


Duration of β-adrenoceptor blockade associated with once-daily oral administration of atenolol in healthy dogs.
OBJECTIVE To test the hypothesis that once-daily oral administration of atenolol would attenuate the heart rate response to isoproterenol for 24 hours. ANIMALS 20 healthy dogs. PROCEDURES A double-blind randomized placebo-controlled crossover study was conducted. Dogs were assigned to receive atenolol (1 mg/kg, PO, q 24 h) or a placebo for 5 to 7 days. After a washout period of 7 days, dogs then received the other treatment. Heart rate at rest (HRr) and heart rate induced by administration of isoproterenol (HRi) as a constant rate infusion (0.2 μg/kg/min for 5 to 7 minutes) were obtained by use of ECG 0, 0.25, 3, 6, 12, 18, and 24 hours after administration of the final dose of atenolol or the placebo. A mixed-model ANOVA was used to evaluate effects of treatment, time after drug or placebo administration, treatment-by-time interaction, period, and sequence on HRr and HRi. RESULTS Effects of sequence or period were not detected. There was a significant effect of treatment and the treatment-by-time interaction on HRi. Atenolol significantly attenuated HRi for 24 hours but did so maximally at 3 hours (least squares mean ± SE, 146 ± 5 beats/min and 208 ± 5 beats/min for atenolol and placebo, respectively). The effect at 24 hours was small (193 ± 5 beats/min and 206 ± 5 beats/min for atenolol and placebo, respectively). Atenolol had a small but significant effect on HRr. CONCLUSIONS AND CLINICAL RELEVANCE This study of healthy dogs receiving atenolol supported a recommendation for a dosing interval < 24 hours.


Bilateral step length estimation using a single inertial measurement unit attached to the pelvis.
The estimation of the spatio-temporal gait parameters is of primary importance in both physical activity monitoring and clinical contexts. A method for estimating step length bilaterally, during level walking, using a single inertial measurement unit (IMU) attached to the pelvis is proposed. In contrast to previous studies, based either on a simplified representation of the human gait mechanics or on a general linear regressive model, the proposed method estimates the step length directly from the integration of the acceleration along the direction of progression. The IMU was placed at pelvis level fixed to the subject's belt on the right side. The method was validated using measurements from a stereo-photogrammetric system as a gold standard on nine subjects walking ten laps along a closed loop track of about 25 m, varying their speed. For each loop, only the IMU data recorded in a 4 m long portion of the track included in the calibrated volume of the SP system, were used for the analysis. The method takes advantage of the cyclic nature of gait and it requires an accurate determination of the foot contact instances. A combination of a Kalman filter and of an optimally filtered direct and reverse integration applied to the IMU signals formed a single novel method (Kalman and Optimally filtered Step length Estimation - KOSE method). A correction of the IMU displacement due to the pelvic rotation occurring in gait was implemented to estimate the step length and the traversed distance. The step length was estimated for all subjects with less than 3% error. Traversed distance was assessed with less than 2% error. The proposed method provided estimates of step length and traversed distance more accurate than any other method applied to measurements obtained from a single IMU that can be found in the literature. In healthy subjects, it is reasonable to expect that, errors in traversed distance estimation during daily monitoring activity would be of the same order of magnitude of those presented.


The translation elongation factor eEF1A1 couples transcription to translation during heat shock response.
Translation elongation factor eEF1A has a well-defined role in protein synthesis. In this study, we demonstrate a new role for eEF1A: it participates in the entire process of the heat shock response (HSR) in mammalian cells from transcription through translation. Upon stress, isoform 1 of eEF1A rapidly activates transcription of HSP70 by recruiting the master regulator HSF1 to its promoter. eEF1A1 then associates with elongating RNA polymerase II and the 3'UTR of HSP70 mRNA, stabilizing it and facilitating its transport from the nucleus to active ribosomes. eEF1A1-depleted cells exhibit severely impaired HSR and compromised thermotolerance. In contrast, tissue-specific isoform 2 of eEF1A does not support HSR. By adjusting transcriptional yield to translational needs, eEF1A1 renders HSR rapid, robust, and highly selective; thus, representing an attractive therapeutic target for numerous conditions associated with disrupted protein homeostasis, ranging from neurodegeneration to cancer.


Changes in subjective wellbeing of prisoners on remand.
Low levels of subjective wellbeing in prisoners may relate to mental health problems and difficulties in reintegration after imprisonment. The development of subjective wellbeing during imprisonment is mostly unclear. The purpose of this paper is to explore this development in a longitudinal study in association with mental disorders and socioeconomic factors. Subjective wellbeing was assessed via a visual analogue scale and retrieved at admission to remand prison and then again after four and eight weeks. Changes in subjective wellbeing between time-points were analyzed taking into account mental disorders and socioeconomic factors, which were assessed by use of the Mini International Neuropsychiatric Interview - Plus and the Camberwell Assessment of Need - Forensic Version, respectively. On average, subjective wellbeing declined directly after remand prison admission, but differences between individuals were found. At remand prison admission, subjective wellbeing significantly improved rather than declined in prisoners with alcohol and substance use disorders, housing problems, unemployment prior to incarceration and in relatively older prisoners. Other related factors did not add significance to this model. In contrast, during remand imprisonment subjective wellbeing displayed an overall increase. For this increase, no predicting factors were found. However, prisoners with an antisocial personality disorder are more at risk of experiencing a decrease in wellbeing during remand imprisonment. In general, the Dutch prison system appears not to result in a decrease in subjective wellbeing in prisoners suffering from a mental disorder during remand imprisonment.


Advancing structured decision-making in drug regulation at the FDA and EMA.
The recent benefit-risk framework (BRF) developed by the Food and Drug Administration (FDA) is intended to improve the clarity and consistency in communicating the reasoning behind the FDA's decisions, acting as an important advancement in US drug regulation. In the PDUFA VI implementation plan, the FDA states that it will continue to explore more structured or quantitative decision analysis approaches; however, it restricts their use within the current BRF that is purely qualitative. By contrast, European regulators and researchers have been long exploring the use of quantitative decision analysis approaches for evaluating drug benefit-risk balance. In this paper, we show how quantitative modelling, backed by decision theory, could complement and extend the FDA's BRF to better support the appraisal of evidence and improve decision outcomes. After providing relevant scientific definitions for benefit-risk assessment and describing the FDA and European Medicines Agency (EMA) frameworks, we explain the components of and differences between qualitative and quantitative approaches. We present lessons learned from the EMA experience with the use of quantitative modelling and we provide evidence of its benefits, illustrated by a real case study that helped to resolve differences of judgements among EMA regulators.


Successes and challenges of HIV prevention in men who have sex with men.
Men who have sex with men (MSM) have been substantially affected by HIV epidemics worldwide. Epidemics in MSM are re-emerging in many high-income countries and gaining greater recognition in many low-income and middle-income countries. Better HIV prevention strategies are urgently needed. Our review of HIV prevention strategies for MSM identified several important themes. At the beginning of the epidemic, stand-alone behavioural interventions mostly aimed to reduce unprotected anal intercourse, which, although somewhat efficacious, did not reduce HIV transmission. Biomedical prevention strategies reduce the incidence of HIV infection. Delivery of barrier and biomedical interventions with coordinated behavioural and structural strategies could optimise the effectiveness of prevention. Modelling suggests that, with sufficient coverage, available interventions are sufficient to avert at least a quarter of new HIV infections in MSM in diverse countries. Scale-up of HIV prevention programmes for MSM is difficult because of homophobia and bias, suboptimum access to HIV testing and care, and financial constraints.


UDP-glucose pyrophosphorylase is not rate limiting, but is essential in Arabidopsis.
UDP-glucose pyrophosphorylase (UGPase) produces UDP-glucose which is essential for sucrose and polysaccharide synthesis. Using Arabidopsis, we demonstrated that two UGPase genes (UGP1 and UGP2) are differentially expressed in a variety of organs, with UGP1 being pre-dominant. Co-expression analyses of UGP genes suggest that UGP1 is closely co-regulated with carbohydrate metabolism genes, late embryogenesis and seed loading, while UGP2 is co-regulated with stress response genes, fertilized flowers and photosynthetic genes. We have used Arabidopsis mutants for the UGP genes to characterize the role of both genes. The UGPase activity/protein was reduced by 70, 10 and 85% in ugp1, ugp2 and ugp1/ugp2 double mutant (DK) plants, respectively. A decrease in UGPase activity/protein was accompanied by an increase in expression of USP, a gene for UDP-sugar pyrophosphorylase, suggesting a compensatory mechanism. Generally, the mutants had no effects on soluble sugar/starch content (except in certain cases for DK plants), and there were no differences in cell wall composition/content between the wild type and the mutants. On the other hand, DK plants had greater hypocotyl and root lengths. When grown in the field, the mutants had as much as a 50% decrease in the number of seeds produced (consistent with a substantial decrease in field fitness), suggesting that they would be outcompeted in the field in a few generations. Overall, the data suggest that UGPase is not rate limiting for sucrose/starch and cell wall synthesis, but that it is essential in Arabidopsis.


Epigenetic regulation of lateralized fetal spinal gene expression underlies hemispheric asymmetries.
Lateralization is a fundamental principle of nervous system organization but its molecular determinants are mostly unknown. In humans, asymmetric gene expression in the fetal cortex has been suggested as the molecular basis of handedness. However, human fetuses already show considerable asymmetries in arm movements before the motor cortex is functionally linked to the spinal cord, making it more likely that spinal gene expression asymmetries form the molecular basis of handedness. We analyzed genome-wide mRNA expression and DNA methylation in cervical and anterior thoracal spinal cord segments of five human fetuses and show development-dependent gene expression asymmetries. These gene expression asymmetries were epigenetically regulated by miRNA expression asymmetries in the TGF-β signaling pathway and lateralized methylation of CpG islands. Our findings suggest that molecular mechanisms for epigenetic regulation within the spinal cord constitute the starting point for handedness, implying a fundamental shift in our understanding of the ontogenesis of hemispheric asymmetries in humans.


Risk profiles for non-communicable diseases in rural and urban schoolchildren in the Republic of Cameroon.
In developing countries, there is evidence that the median age of the population and the life expectancy at birth are increasing as a result of decreasing fertility rates and infant mortality. The result is an aging population more prone to non-communicable diseases such as diabetes, cancer, or heart disease later in life. In addition, changing lifestyle factors such as tobacco use, physical inactivity, and high fat diets, may accelerate the emergence of such chronic diseases as major causes of death and disability in these countries, particularly in urban areas. To test the premise that urban living predisposes residents to reduced activity levels, less healthy diets, cigarette smoking, elevated blood pressure, and increased body fat early in life, we studied rural/urban differences in these risk factors among schoolchildren in the Republic of Cameroon. One hundred and nineteen Class 7 schoolchildren (50 urban and 69 rural) were interviewed concerning diet, physical activity, smoking, and alcohol use; blood pressure and anthropometric measurements were also taken. Physical activity among rural children was more than twice that of urban children, and most of the activity for rural children was work-related. Rural children consumed fewer foods containing fat and more fruits and vegetables. Adjusting for age, systolic and diastolic blood pressures of urban boys were higher than those of rural boys, and among urban children there was a trend toward a larger age-adjusted mean body mass index (BMI). There were no differences in alcohol or tobacco use between urban and rural children. In this study, urbanization was associated with a less active lifestyle and a dietary pattern that was higher in fat and lower in fruit and vegetable intake. Since risk factors for non-communicable diseases tend to appear early in life and track into adulthood, it is important to identify those children, or groups of children, with unfavorable risk profiles and to structure health education and promotion programs to modify these trends.


Assembly of adeno-associated virus type 2 capsids in vitro.
Capsid proteins VP1, VP2 and VP3 of adeno-associated virus type 2 (AAV-2) were separately expressed by recombinant baculoviruses, purified under denaturing conditions and renatured in the presence of 0.5 M arginine, followed by dialysis against buffers of physiological ionic strength. At a protein concentration of 0.05 mg/ml, the three capsid proteins predominantly formed monomers and, to a lesser extent, oligomers, as determined by sedimentation analysis. Oligomerization increased at higher protein concentrations. The capsid protein oligomers consisted of globular, non-capsid-like structures, as detected by electron microscopy. Addition of a HeLa cell extract significantly stimulated oligomerization of the capsid proteins, probably due to interactions with HeLa cell proteins. Characterization of structures sedimenting around 60S by immunoprecipitation and electron microscopy showed that, in addition to other aggregates, empty capsid-like structures were formed in vitro. The identity of these structures as empty AAV capsids was verified by immunoelectron microscopy. Analysis of capsid formation in HeLa cells by transfection of VP expression constructs allowing separate expression of VP1, VP2 and VP3 showed that they were able to form capsids, although with a reduced efficiency as compared to VP proteins expressed from the wt cap gene. This finding suggests that the mutations introduced to allow separate capsid protein expression reduced the efficiency of capsid assembly in vivo and might also explain the reduced recovery of empty capsids employing the in vitro assembly procedure.


Distinct pathological subtypes of FTLD-FUS.
Most cases of frontotemporal lobar degeneration (FTLD) are characterized by abnormal intracellular accumulation of either tau or TDP-43 protein. However, in ~10% of cases, composed of a heterogenous collection of uncommon disorders, the molecular basis remains to be uncertain. We recently discovered that the pathological changes in several tau/TDP-43-negative FTLD subtypes are immunoreactive (ir) for the fused in sarcoma (FUS) protein. In this study, we directly compared the pattern of FUS-ir pathology in cases of atypical FTLD-U (aFTLD-U, N = 10), neuronal intermediate filament inclusion disease (NIFID, N = 5) and basophilic inclusion body disease (BIBD, N = 8), to determine whether these are discrete entities or represent a pathological continuum. All cases had FUS-ir pathology in the cerebral neocortex, hippocampus and a similar wide range of subcortical regions. Although there was significant overlap, each group showed specific differences that distinguished them from the others. Cases of aFTLD-U consistently had less pathology in subcortical regions. In addition, the neuronal inclusions in aFTLD-U usually had a uniform, round shape, whereas NIFID and BIBD were characterized by a variety of inclusion morphologies. In all cases of aFTLD-U and NIFID, vermiform neuronal intranuclear inclusions (NII) were readily identified in the hippocampus and neocortex. In contrast, only two cases of BIBD had very rare NII in a single subcortical region. These findings support aFTLD-U, NIFID and BIBD as representing closely related, but distinct entities that share a common molecular pathogenesis. Although cases with overlapping pathology may exist, we recommend retaining the terms aFTLD-U, NIFID and BIBD for specific FTLD-FUS subtypes.


Highly sensitive and broadly reactive quantitative reverse transcription-PCR assay for high-throughput detection of Rift Valley fever virus.
Rift Valley fever (RVF) virus is a mosquito-borne virus associated with large-scale epizootics/epidemics throughout Africa and the Arabian peninsula. Virus infection can result in economically disastrous "abortion storms" and high newborn mortality in livestock. Human infections result in a flu-like illness, with 1 to 2% of patients developing severe complications, including encephalitis or hemorrhagic fever with high fatality rates. There is a critical need for a highly sensitive and specific molecular diagnostic assay capable of detecting the natural genetic spectrum of RVF viruses. We report here the establishment of a pan-RVF virus quantitative real-time reverse transcription-PCR assay with high analytical sensitivity (approximately 5 RNA copies of in vitro-transcribed RNA/reaction or approximately 0.1 PFU of infectious virus/reaction) and efficiency (standard curve slope = -3.66). Based on the alignments of the complete genome sequences of 40 ecologically and biologically diverse virus isolates collected over 56 years (1944 to 2000), the primer and probe annealing sites targeted in this assay are known to be located in highly conserved genomic regions. The performance of this assay relative to serologic assays is illustrated by testing of known RVF case materials obtained during the Saudi Arabia outbreak in 2000. Furthermore, analysis of acute-phase blood samples collected from human patients (25 nonfatal, 8 fatal) during that outbreak revealed that patient viremia at time of presentation at hospital may be a useful prognostic tool in determining patient outcome.


Recent methods for the synthesis of α-acyloxy ketones.
The present review provides a broad description of the methods reported for the synthesis of α-acyloxy ketones and some of their derivatives. α-Acyloxy ketones and their derivatives are vital synthetic intermediates and are ubiquitous as biochemical constituents of living organisms, biologically active natural products and pharmaceuticals. Due to their biological importance, new methods for their synthesis are being continuously developed and documented in the recent past. However, the chemical literature lacks a comprehensive summary on the synthetic methodologies targeting α-acyloxy ketones. In an attempt to fill this void, this review discusses their synthetic procedures developed over time. The synthetic approaches are systematically classified based on the substrates used. The mechanistic details for certain critical cases are also discussed. In the past, preparation of α-acyloxy ketones was reported from functionalized ketones like α-haloketones and diazo esters. Later on, among the reactions that formed the acyloxy ketones, oxidative coupling of ketones with carboxylic acids both under metal and metal-free conditions made their synthesis simple and versatile. Specifically, in the last decade, many oxidative coupling reactions emerged as a powerful tool for the synthesis of α-acyloxy ketones. Quite recently, acyloxy ketones' synthesis has been reported from commercially available alkenes and alkynes through oxidative addition reactions. Subsequently, the mechanistic details for these coupling reactions became interesting to many organic chemists. The asymmetric version of the title compounds hails from their enzymatic resolution to metal catalysed chiral synthesis. Besides, the synthesis of acyloxy ketones from epoxides, alcohols and enamides using various oxidative reagents has also been documented.


Patients' associations and the control of leishmaniasis in Peru.
American mucocutaneous leishmaniasis is an important health problem in Peru, particularly in the mountainous Cuzco Region, where 25% of all new cases reported in 1989 were located. Cases have increased considerably since the beginning of the 1980s, when large-scale seasonal migration to endemic zones occurred, particularly the forest area of Madre de Dios, following the discovery of new gold deposits there, and the deterioration in the economic situation in Peru. Following the lack of official response from the Peruvian government, hundreds of people suffering from leishmaniasis in the Cuzco area formed self-help associations with the objective of obtaining the drugs needed to treat their disease. The major achievement of this spontaneous movement, which was supported by several public and private institutions, was to encourage sick people, particularly patients with mucosal lesions, to emerge from isolation. As a result, the prevalence and incidence of the disease have now considerably decreased in the region.


Double transseptal puncture for catheter ablation of atrial fibrillation: safety of the technique and its use in the outpatient setting.
Introduction. For pulmonary vein isolation in patients with atrial fibrillation (AF), some centers use the double transseptal puncture technique for catheter access in order to facilitate catheter manipulation within the left atrium. However, no safety data has so far been published using this approach. Method. 269 ablation procedures were performed in 243 patients (mean age 56.6 ± 9.3 years, 75% men) using the double transseptal puncture for catheter access in all cases. Patients were considered for ablation of paroxysmal (80%), persistent (19%), and permanent (1%) AF. 230 procedures were performed on an outpatient basis (85.5%), and 26 were repeat procedures (9.7%). Results. The double transseptal puncture catheter access was successfully achieved in all patients. The procedural success with the endpoint of pulmonary vein isolation was reached in 255 procedures (95%). A total of 1048 out of 1062 pulmonary veins (99%) were successfully isolated. Major complications occurred in eight patients (3.0%). Of these, seven patients (2.6%) had pericardial effusion requiring percutaneous drainage, and one patient (0.4%) suffered a minor reversible stroke. One patient (0.4%) had a minor air embolism with transient symptoms. Conclusion. The double transseptal puncture catheterization technique allows easy catheter manipulation within the left atrium to reach the goal of acute procedural success in AF ablation. Procedure-related complications are rare, and the technique can be used safely for AF ablation in the outpatient setting.


Probing Molecular Excited States by Atomic Force Microscopy.
By employing single charge injections with an atomic force microscope, we investigated redox reactions of a molecule on a multilayer insulating film. First, we charged the molecule positively by attaching a single hole. Then we neutralized it by attaching an electron and observed three channels for the neutralization. We rationalize that the three channels correspond to transitions to the neutral ground state, to the lowest energy triplet excited states and to the lowest energy singlet excited states. By single-electron tunneling spectroscopy we measured the energy differences between the transitions obtaining triplet and singlet excited state energies. The experimental values are compared with density functional theory calculations of the excited state energies. Our results show that molecules in excited states can be prepared and that energies of optical gaps can be quantified by controlled single-charge injections. Our work demonstrates the access to, and provides insight into, ubiquitous electron-attachment processes related to excited-state transitions important in electron transfer and molecular optoelectronics phenomena on surfaces.


Altered pharyngeal muscles in Parkinson disease.
Dysphagia (impaired swallowing) is common in patients with Parkinson disease (PD) and is related to aspiration pneumonia, the primary cause of death in PD. Therapies that ameliorate the limb motor symptoms of PD are ineffective for dysphagia. This suggests that the pathophysiology of PD dysphagia may differ from that affecting limb muscles, but little is known about potential neuromuscular abnormalities in the swallowing muscles in PD. This study examined the fiber histochemistry of pharyngeal constrictor and cricopharyngeal sphincter muscles in postmortem specimens from 8 subjects with PD and 4 age-matched control subjects. Pharyngeal muscles in subjects with PD exhibited many atrophic fibers, fiber type grouping, and fast-to-slow myosin heavy chain transformation. These alterations indicate that the pharyngeal muscles experienced neural degeneration and regeneration over the course of PD. Notably, subjects with PD with dysphagia had a higher percentage of atrophic myofibers versus with those without dysphagia and controls. The fast-to-slow fiber-type transition is consistent with abnormalities in swallowing, slow movement of food, and increased tone in the cricopharyngeal sphincter in subjects with PD. The alterations in the pharyngeal muscles may play a pathogenic role in the development of dysphagia in subjects with PD.


Odd-electron molecular theory of graphene hydrogenation.
This paper highlights the molecular essence of graphene and presents its hydrogenation from the viewpoint of the odd-electron molecular theory. This chemical transformation was performed computationally, using a particular algorithm, through the stepwise addition of either hydrogen molecules or hydrogen atoms to a pristine graphene molecule. The graphene was considered to be a membrane, such that either both sides or just one side of the membrane was accessible to adsorbate, and the atoms on the perimeter of the membrane were either fixed (fixed membrane) or free to move (free-standing membrane). The algorithm explored the spatial distribution of the number of effectively unpaired electrons N (DA) over the carbon skeleton of the molecule. The highest ranked N (DA) values were considered to indicate the target atoms at each reaction step. The dependence of the hydrogenation itself and the final graphene hydrides on external factors such as whether the membrane was fixed, if both sides or only one side of the membrane were accessible to hydrogen, and whether the hydrogen was in the molecular or atomic state. Complete hydrogenation followed by the formation of a regular chairlike graphane structure (CH)(n) was only found to be possible for a fixed pristine graphene membrane for which the basal plane is accessible to hydrogen atoms from both sides.


Impaired human hippocampal neurogenesis after treatment for central nervous system malignancies.
The effects of cancer treatments such as cranial radiation and chemotherapy on human hippocampal neurogenesis remain unknown. In this study, we examine neuropathological markers of neurogenesis and inflammation in the human hippocampus after treatment for acute myelogenous leukemia or medulloblastoma. We demonstrate a persistent radiation-induced microglial inflammation that is accompanied by nearly complete inhibition of neurogenesis after cancer treatment. These findings are consistent with preclinical animal studies and suggest potential therapeutic strategies.


Hydrostatic pressure promotes the proliferation and osteogenic/chondrogenic differentiation of mesenchymal stem cells: The roles of RhoA and Rac1.
Our previous studies have shown that hydrostatic pressure can serve as an active regulator for bone marrow mesenchymal stem cells (BMSCs). The current work further investigates the roles of cytoskeletal regulatory proteins Ras homolog gene family member A (RhoA) and Ras-related C3 botulinum toxin substrate 1 (Rac1) in hydrostatic pressure-related effects on BMSCs. Flow cytometry assays showed that the hydrostatic pressure promoted cell cycle initiation in a RhoA- and Rac1-dependent manner. Furthermore, fluorescence assays confirmed that RhoA played a positive and Rac1 displayed a negative role in the hydrostatic pressure-induced F-actin stress fiber assembly. Western blots suggested that RhoA and Rac1 play central roles in the pressure-inhibited ERK phosphorylation, and Rac1 but not RhoA was involved in the pressure-promoted JNK phosphorylation. Finally, real-time polymerase chain reaction (PCR) experiments showed that pressure promoted the expression of osteogenic marker genes in BMSCs at an early stage of osteogenic differentiation through the up-regulation of RhoA activity. Additionally, the PCR results showed that pressure enhanced the expression of chondrogenic marker genes in BMSCs during chondrogenic differentiation via the up-regulation of Rac1 activity. Collectively, our results suggested that RhoA and Rac1 are critical to the pressure-induced proliferation and differentiation, the stress fiber assembly, and MAPK activation in BMSCs.


Penicillin in oral aphthosis, new insight for an old drug: A randomized, double-blind, controlled clinical trial.
Oral aphthosis is a painful ulceration of mucus membranes characterized by round or oval lesions with central necrosis and erythematous haloes. Due to unknown etiology, treatment is highly controversial and based mainly on individual experience. The aim of this study was to investigate the safety and efficacy of topical penicillin 6.3.3 for the treatment of recurrent aphthous stomatitis. This randomized, double-blind, controlled clinical trial was done in Shahid Sadoughi Hospital Clinic in Yazd (2011-2012). Fifty patients aged 15-45 with recurrent oral aphthosis were randomly divided into two groups. After obtaining informed consents, patients in the case and control groups were treated (four times/day for a week), respectively, by topical penicillin 6.3.3 powder and placebo in similar vial. The patients who had acute-onset oral aphthae (≤48 h of appearance) with diameter ≥5 mm were included. History of sensitivity to β-lactam antibiotics and cephalosporin; spontaneous recovery during <5 days in previous episodes; concurrent systemic, infectious, or any autoimmune disorders; history of taking drugs (local or systemic) from 2 weeks prior to presentation; alcohol or drug abuse; smoking cigarette or tobacco; and poor compliance were exclusion criteria. Patients were examined in days 0, 3, 6, and 8. The main outcome measure was reduction in the median pain. Burning, pain, erythema, and inflammation were recorded as complications. Of 25 patients receiving penicillin, 13 were female and 12 were male. Regarding the pain score (mean difference = 1.6 vs. 0.88, P = 0.012) and size of aphthus (mean difference = 9.43 vs. 1.24, P = 0.008), patients who received penicillin had significantly better results than the placebo group on day 8 after the treatment. The mean duration to healing was 3 days for penicillin group and 6 days for placebo group (P = 0.016). No topical or systemic adverse effects were observed. Our study showed a dramatic response to topical penicillin with respect to placebo. Hence, it seems that penicillin could be a safe and effective option in managing oral aphthosis.


Mapping the Azolog Space Enables the Optical Control of New Biological Targets.
Photopharmacology relies on molecules that change their biological activity upon irradiation. Many of these are derived from known drugs by replacing their core with an isosteric azobenzene photoswitch (azologization). The question is how many of the known bioactive ligands could be addressed in such a way. Here, we systematically assess the space of molecules amenable to azologization from databases of bioactive molecules (DrugBank, PDB, CHEMBL) and the Cambridge Structural Database. Shape similarity scoring functions (3DAPfp) and analyses of dihedral angles are employed to quantify the structural homology between a bioactive molecule and the cis or trans isomer of its corresponding azolog ("azoster") and assess which isomer is likely to be active. Our analysis suggests that a very large number of bioactive ligands (>40 000) is amenable to azologization and that many new biological targets could be addressed with photopharmacology. N-Aryl benzamides, 1,2-diarylethanes, and benzyl phenyl ethers are particularly suited for this approach, while benzylanilines and sulfonamides appear to be less well-matched. On the basis of our analysis, the majority of azosters are expected to be active in their trans form. The broad applicability of our approach is demonstrated with photoswitches that target a nuclear hormone receptor (RAR) and a lipid processing enzyme (LTA4 hydrolase).


Exposure of early pediatric trainees to blood and marrow transplantation leads to higher recruitment to the field.
The National Marrow Donor Program (NMDP) projects the need for allogeneic unrelated blood and marrow transplantation (BMT) in the United States as 10,000 per year. Although the NMDP is preparing to facilitate that number by the year 2015, there are several barriers to meeting this goal, including the need to recruit more health care personnel, including BMT physicians. To learn how best to recruit BMT physicians, we examined why practicing BMT physicians chose to enter the field and why others did not. We conducted a Web-based survey among pediatric hematology/oncology (PHO) and BMT physician providers and trainees to identify the factors influencing their decision to choose or not choose a career in BMT. Out of 259 respondents (48% male, 74% of Caucasian origin), 94 self-identified as BMT physicians, 112 as PHO physicians, and 53 as PHO trainees. The PHO and BMT providers spent an average of 53% of their time in clinical activities. More than two-thirds of PHO providers reported providing BMT services at their institutions, most commonly for inpatient coverage (73%). The proportion of providers exposed to BMT early in training was significantly higher among BMT providers compared with PHO providers (51% versus 18% in medical school [P < .0001]; 70% versus 50% during residency [P < .005]). Exposure during fellowship (94%) did not differ between the 2 groups. The decision to pursue a career in BMT was made before fellowship (medical school or residency) by 50% of the respondents. A lower proportion of BMT providers than PHO providers reported current involvement in the education of medical students and residents (76% versus 98%; P < .0001). Of the 53 trainees who responded, 64% reported not contemplating a career in BMT. Of these, 68% identified inadequate exposure to BMT before PHO fellowship as the reason behind this decision. Only 26% reported receiving exposure to the BMT field while in medical school, and 43% reported exposure during residency. The 2 most common reasons cited for choosing a career as a BMT physician were the degree of intellectual and scientific challenge (89%) and the influence of role models/mentors in the field (67%). The results of this survey suggest that early exposure to BMT during medical school and residency is associated with increased interest in pursuing a career in BMT. BMT physicians and training program directors can foster interest in the field by promoting BMT-focused education and clinical inpatient and outpatient rotations during medical school and residency. This early exposure to BMT may aid recruitment of future transplantation providers.


Next generation sequencing for TCR repertoire profiling: platform-specific features and correction algorithms.
The TCR repertoire is a mirror of the human immune system that reflects processes caused by infections, cancer, autoimmunity, and aging. Next generation sequencing (NGS) is becoming a powerful tool for deep TCR profiling; yet, questions abound regarding the methodological approaches for sample preparation and correct data interpretation. Accumulated PCR and sequencing errors along with library preparation bottlenecks and uneven PCR efficiencies lead to information loss, biased quantification, and generation of huge artificial TCR diversity. Here, we compare Illumina, 454, and Ion Torrent platforms for individual TCR profiling, evaluate the rate and character of errors, and propose advanced platform-specific algorithms to correct massive sequencing data. These developments are applicable to a wide variety of next generation sequencing applications. We demonstrate that advanced correction allows the removal of the majority of artificial TCR diversity with concomitant rescue of most of the sequencing information. Thus, this correction enhances the accuracy of clonotype identification and quantification as well as overall TCR diversity measurements.


Fibronectin-binding protein A of Staphylococcus aureus has multiple, substituting, binding regions that mediate adherence to fibronectin and invasion of endothelial cells.
Invasive Staphylococcus aureus infection frequently involves bacterial seeding from the bloodstream to other body tissues, a process necessarily involving interactions between circulating bacteria and vascular endothelial cells. Staphylococcus aureus fibronectin-binding protein is central to the invasion of endothelium, fibronectin forming a bridge between bacterial fibronectin-binding proteins and host cell receptors. To dissect further the mechanisms of invasion of endothelial cells by S. aureus, a series of truncated FnBPA proteins that lacked one or more of the A, B, C or D regions were expressed on the surface of S. aureus and tested in fibronectin adhesion, endothelial cell adhesion and invasion assays. We found that this protein has multiple, substituting, fibronectin-binding regions, each capable of conferring both adherence to fibronectin and endothelial cells, and endothelial cell invasion. By expressing S. aureus FnBPA on the surface of the non-invasive Gram-positive organism Lactococcus lactis, we have found that no other bacterial factor is required for invasion. Furthermore, we have demonstrated that, as with other cell types, invasion of endothelial cells is mediated by integrin alpha5beta1. These findings may be of relevance to the development of preventive measures against systemic infection, and bacterial spread in the bacteraemic patient.


Mast cell-mediated long-lasting increases in excitability of vagal C fibers in guinea pig esophagus.
Several esophageal pathologies are associated with an increased number of mast cells in the esophageal wall. We addressed the hypothesis that activation of esophageal mast cells leads to an increase in the excitability of local sensory C fibers. Guinea pigs were actively sensitized to ovalbumin. The mast cells in the esophagus were selectively activated ex vivo by superfusion with ovalbumin. Action potential discharge in guinea pig vagal nodose esophageal C-fiber nerve endings was monitored in the isolated (ex vivo) vagally innervated esophagus by extracellular recordings. Ovalbumin activated esophageal mast cells, leading to the rapid release of approximately 20% of the tissue histamine stores. This was associated with a consistent and significant increase in excitability of the nodose C fibers as reflected in a two- to threefold increase in action potential discharge frequency evoked by mechanical (increases in intraluminal pressure) stimulation. The increase in excitability persisted unchanged for at least 90 min (longest time period tested) after ovalbumin was washed from the tissue. This effect could be prevented by the histamine H1 receptor antagonist pyrilamine, but once the increase in excitability occurred, it persisted in the nominal absence of histamine and could not be reversed even with large concentrations of the histamine receptor antagonist. In conclusion, activation of esophageal mast cells leads to a pronounced and long-lived increase in nociceptive C-fiber excitability such that any sensation or reflex evoked via the vagal nociceptors will likely be enhanced. The effect is initiated by histamine acting via H1 receptor activation and maintained in the absence of the initiating stimulus.


Real-time computer simulations of excitable media: JAVA as a scientific language and as a wrapper for C and FORTRAN programs.
We describe a useful setting for interactive, real-time study of mathematical models of cardiac electrical activity, using implicit and explicit integration schemes implemented in JAVA. These programs are intended as a teaching aid for the study and understanding of general excitable media. Particularly for cardiac cell models and the ionic currents underlying their basic electrical dynamics. Within the programs, excitable media properties such as thresholds and refractoriness and their dependence on parameter values can be analyzed. In addition, the cardiac model applets allow the study of reentrant tachyarrhythmias using premature stimuli and conduction blocks to induce or to terminate reentrant waves of electrical activation in one and two dimensions. The role of some physiological parameters in the transition from tachycardia to fibrillation also can be analyzed by varying the maximum conductances of ion channels associated with a given model in real time during the simulations. These applets are available for download at http://arrhythmia.hofstra.edu or its mirror site http://stardec.ascc.neu.edu/~fenton.