Hypertrophic cardiomyopathy: The need for randomized trials.
Hypertrophic cardiomyopathy (HCM) is a complex cardiac condition characterized by variable degrees of asymmetric left ventricular (LV) hypertrophy, generally associated with mutations in sarcomere protein genes. While generally perceived as rare, HCM is the most common genetic heart disease with over one million affected individuals in Europe alone and represents a prevalent cause of sudden cardiac death in the young. To date, HCM remains an orphan disease, as recommended treatment strategies are based on the empirical use of old drugs with little evidence supporting their clinical benefit in this context. In the six decades since the original description of the disease, less than fifty pharmacological studies have been performed in HCM patients, enrolling little over 2,000 HCM patients, mostly comprising small non-randomized cohorts. No specific agent has been convincingly shown to affect outcome, and critical issues such as prevention of myocardial energy depletion, microvascular ischemia, progressive myocardial fibrosis and the peculiar mechanisms of arrhythmogenesis in HCM still need to be addressed in a systematic fashion. However, there is increasing evidence that a variety of drugs may counter the effects of sarcomere protein mutations and the resulting pathophysiological abnormalities at the molecular, cellular and organ level. Following major advances in our understanding of HCM and increasing opportunities for networking among large international referral centres, the opportunity now exists to identify potentially effective treatments and implement adequately designed pharmacological trials, with the ultimate aim to impact the natural course of the disease, alleviate symptoms and improve quality of life in our patients.


Isolation and characterization of monoclonal antibodies raised against the reverse transcriptase of human immunodeficiency virus type 2 and cross-reactivity with that of type 1.
Monoclonal antibodies to human immunodeficiency virus (HIV)-2 reverse transcriptase have been raised with the ultimate goal of generating Fab fragments for future co-crystallization studies. A number of mouse monoclonal antibodies to recombinant HIV-2 reverse transcriptase have been obtained and characterized in terms of the possible epitopes they recognise together with cross-reactivity with a related reverse transcriptase. The antibodies were shown to fall into three groups that recognize different regions of the reverse transcriptase enzyme. One antibody, which recognizes part of the RNase H domain, demonstrated cross-reactivity between the HIV-1 and HIV-2 reverse transcriptase.


Posttranslational regulation of insulin-like growth factor binding protein-4 in normal and transformed human fibroblasts. Insulin-like growth factor dependence and biological studies.
Insulin-like growth factor binding protein-4 (IGFBP-4) is a 24-26-kD protein expressed by a variety of cell types in vivo and in vitro. Treatment of normal adult human fibroblasts with 10 nM insulin-like growth factor II (IGF-II) for 24 h resulted in an 85% decrease in endogenous IGFBP-4, as assessed by Western ligand blot analysis of the conditioned medium. Incubation of human fibroblast-conditioned medium (HFCM) with IGF-II under cell-free conditions led to a similar loss of IGFBP-4. This posttranslationally regulated decrease in IGFBP-4 appeared to be due to a protease in HFCM: (a) It could be prevented with specific protease inhibitors or incubation at 4 degrees C; (b) proteolysis of recombinant human (rh) IGFBP-4 required HFCM; (c) immunoblotting and radiolabeling confirmed cleavage of IGFBP-4 into 18- and 14-kD IGFBP-4 fragments. The protease was specific for IGFBP-4, and was strictly dependent on IGFs for activation. IGF-II was the most effective of the natural and mutant IGFs tested, inducing complete hydrolysis of rhIGFBP-4 at a molar ratio of 0.25:1 (IGF/IGFBP-4). Simian virus 40-transformed adult human fibroblasts also expressed IGFBP-4 and IGFBP-4 protease, as well as an inhibitor of IGFBP-4 proteolysis. In biological studies, intact rhIGFBP-4 potently inhibited IGF-I-stimulated [3H]aminoisobutyric acid uptake, whereas proteolyzed rhIGFBP-4 had no inhibitory effect. In conclusion, these data provide evidence for a novel IGF-dependent IGFBP-4-specific protease that modifies IGFBP-4 structure and function, and indicate a preferential role for IGF-II in its activation. Posttranslational regulation of IGFBP-4 may provide a means for cooperative control of local cell growth by IGF-I and IGF-II.


Once-daily atomoxetine treatment for children with attention-deficit/hyperactivity disorder, including an assessment of evening and morning behavior: a double-blind, placebo-controlled trial.
Atomoxetine seems to be as effective for treating attention-deficit/hyperactivity disorder (ADHD) when the daily dose is administered once in the morning as when the dose is divided and administered in the morning and evening. In the present study, the efficacy of atomoxetine administered once daily among children with ADHD was assessed throughout the day, including the evening and early morning. Another goal was to determine how early in treatment it was possible to discern a specific effect of the drug on ADHD symptoms. This study was a randomized, multicenter, double-blind, placebo-controlled trial conducted at 12 outpatient sites in the United States. A total of 197 children, 6 to 12 years of age, who had been diagnosed as having ADHD, on the basis of the Diagnostic and Statistical Manual of Mental Disorders (4th ed.) criteria, were randomized to receive 8 weeks of treatment with atomoxetine or placebo, dosed once daily in the mornings. ADHD symptoms were assessed with parent and investigator rating scales. The primary outcome measure was the Attention-Deficit/Hyperactivity Disorder Rating Scale-IV-Parent Version: Investigator-Administered and Scored total score. Daily parent assessments of children's home behaviors in the evening and early morning were recorded with an electronic data entry system. This instrument measures 11 specific morning or evening activities, including getting up and out of bed, doing or completing homework, and sitting through dinner. Seventy-one percent of the children enrolled were male, 69% met criteria for the combined subtype (both inattentive and hyperactive/impulsive symptoms), and the most common psychiatric comorbidity was oppositional defiant disorder (35%). Once-daily atomoxetine (final mean daily dose of 1.3 mg/kg) was significantly more effective than placebo in treating core symptoms of ADHD. Mean reductions in the Attention-Deficit/Hyperactivity Disorder Rating Scale-IV-Parent Version: Investigator-Administered and Scored total score were significantly greater for patients randomized to atomoxetine, beginning at the first visit after the initiation of treatment and continuing at all subsequent visits. Both inattentive and hyperactive/impulsive symptom clusters were significantly reduced with atomoxetine, compared with placebo. With continued treatment and dose titrations, core symptoms of ADHD continued to decrease throughout the 8-week study. Mean reductions in the daily parent assessment total scores for patients randomized to atomoxetine were superior during the first week, beginning with the first day of dosing, and were also superior at endpoint. Efficacy outcomes for the evening hours for atomoxetine-treated patients were superior to those for placebo-treated patients, as assessed with 2 different assessment scales. Decreases in the daily parent assessment morning subscores at endpoint showed a significant reduction in symptoms that lasted into the mornings. Rates of discontinuations attributable to adverse events were <5% for both groups. Adverse events reported significantly more frequently with atomoxetine were decreased appetite, somnolence, and fatigue. Among children 6 to 12 of age who had been diagnosed as having ADHD, once-daily administration of atomoxetine in the morning provided safe, rapid, continuous, symptom relief that lasted not only into the evening hours but also into the morning hours. Atomoxetine treatment was safe and well tolerated.


The ultrastructural localization of tri-n-butyltin in human erythrocyte membranes during shape transformation leading to hemolysis.
Scanning electron microscopy, transmission electron microscopy, freeze-fracture, and x-ray energy dispersive spectrometry were used to localize tri-n-butyltin (TBT) in human erythrocytes (RBC's). TBT induced a rapid shape transformation of the RBC discocyte to an echinocyte, which led to hemolysis at concentrations at or above 10 microM TBT. Electron dense spheres or ellipsoids were observed in association with blood cell membranes at or above 10 microM TBT. These structures were visualized initially in thin sections when postfixed with osmium tetroxide. Control cell preparations without TBT did not exhibit these structural densities when fixed with osmium. Freeze-fracture replicas confirmed the presence of TBT aggregates associated with cell membranes as intercalations in the lipid bilayer. In thin sections, these structures measured 71.5 +/- 18.2 nm in diameter. In freeze-fracture replicas of TBT-treated RBC's, particulate structures measuring 60 +/- 18.5 nm in diameter were present on membrane exoplasmic fracture faces and 59.6 +/- 10.8-nm depressions on membrane protoplasmic fracture faces. Qualitative x-ray energy dispersive spectrometry analysis of ultrathin sections of glutaraldehyde-carbohydrazide-embedded samples revealed that the membrane-associated aggregates contained tin. TBT-treated RBC's that were washed with normal saline resulted in a paucity of TBT aggregates associated with the membranes and a reduction in the RBC hemolysis rate. RBC shape transformation occurred at each concentration examined from 0.1 to 100 microM TBT, but was reversible below 1 microM TBT.


Variability in resource use: diagnosing colorectal cancer.
Efficient resource use is relevant in all healthcare systems. Although colorectal cancer is common, little has been published regarding the utilization of clinical resources in diagnosis. The primary aim was to evaluate the patterns and factors associated with clinical services used to diagnose colorectal cancer at 14 US Department of Veterans Affairs facilities. The secondary aim was to investigate whether using more clinical services was associated with time to diagnosis. We reviewed medical records for 449 patients with colorectal cancer in an observational study. Study end points were the use of clinical diagnostic services grouped as laboratory tests, imaging studies, and subspecialty consultations. Cumulative logistic regression models were used to explore factors associated with each outcome. Facility variability contributed to the variability of resource use in all models. In adjusted analyses, older patients had higher use of laboratory tests (odds ratio [OR], 1.20; 95% confidence interval [CI], 1.02-1.43) and incidentally discovered colorectal cancer was associated with increased use of consultations (OR, 1.97; 95% CI, 1.27-3.05), imaging studies (OR, 1.70; 95% CI, 1.12-2.58), and laboratory tests (OR, 3.14; 95% CI, 2.06-4.77) compared with screen-detected cancers. There was a strong direct correlation between thenumber of diagnostic services performed and the median time to diagnosis (Spearman correlation coefficient, 0.99; P < .001). Variability in utilization of diagnostic clinical services was associated with patient age, patient presentation, and facility. Increased resource use was highly correlated with increased time to diagnosis.


Psychosocial factors associated with paternal postnatal depression.
While maternal postpartum depression is a well-known phenomenon, paternal postnatal depression has been less studied. It is known that paternal postnatal depression impacts on children's and families' development, affects marital satisfaction and affects the economic health of industrialized countries. The aim of this study was to identify the psychosocial factors associated with paternal postnatal depression. A descriptive-correlational study was conducted with a sample of fathers of infants (average age: 11 months) who were breastfed exclusively or predominantly for at least 6 months, comparing psychosocial factors in fathers with (n: 17, 8.2%) and without a positive score for depression on the EPDS scale (n: 188). Psychosocial factors were assessed through questionnaires. Depression in fathers of breastfed infants is associated with the experience of perinatal loss in a previous pregnancy, parenting distress, infant temperament (difficult child), dysfunctional interactions with the child, decreased marital adjustment and perceived low parenting efficacy. Multivariate analysis suggests an independent effect of psychosocial factors such as parenting distress, quality of the marital relationship and perceived parenting efficacy on paternal depression. The sample focused on fathers of breastfed infant, since breastfeeding has become the feeding norm, and this should be taken into account when considering the generalization of findings. These findings emphasize the need to consider a set of psychosocial factors when examining fathers' mental health in the first year of a child's birth. Health professionals can enhance parenting efficacy and alleviate parenting distress by supporting fathers' unique experiences and addressing their needs.


Motor imagery in spinal cord injured people is modulated by somatotopic coding, perspective taking, and post-lesional chronic pain.
Motor imagery (MI) allows one to mentally represent an action without necessarily performing it. Importantly, however, MI is profoundly influenced by the ability to actually execute actions, as demonstrated by the impairment of this ability as a consequence of lesions in motor cortices, limb amputations, movement limiting chronic pain, and spinal cord injury. Understanding MI and its deficits in patients with motor limitations is fundamentally important as development of some brain-computer interfaces and daily life strategies for coping with motor disorders are based on this ability. We explored MI in a large sample of patients with spinal cord injury (SCI) using a comprehensive battery of questionnaires to assess the ability to imagine actions from a first-person or a third-person perspective and also imagine the proprioceptive components of actions. Moreover, we correlated MI skills with personality measures and clinical variables such as the level and completeness of the lesion and the presence of chronic pain. We found that the MI deficits (1) concerned the body parts affected by deafferentation and deefferentation, (2) were present in first- but not in third-person perspectives, and (3) were more altered in the presence of chronic pain. MI is thus closely related to bodily perceptions and representations. Every attempt to devise tools and trainings aimed at improving autonomy needs to consider the cognitive changes due to the body-brain disconnection.


Crystal structure of a pol alpha family DNA polymerase from the hyperthermophilic archaeon Thermococcus sp. 9 degrees N-7.
The 2.25 A resolution crystal structure of a pol alpha family (family B) DNA polymerase from the hyperthermophilic marine archaeon Thermococcus sp. 9 degrees N-7 (9 degrees N-7 pol) provides new insight into the mechanism of pol alpha family polymerases that include essentially all of the eukaryotic replicative and viral DNA polymerases. The structure is folded into NH(2)- terminal, editing 3'-5' exonuclease, and polymerase domains that are topologically similar to the two other known pol alpha family structures (bacteriophage RB69 and the recently determined Thermococcus gorgonarius), but differ in their relative orientation and conformation. The 9 degrees N-7 polymerase domain structure is reminiscent of the "closed" conformation characteristic of ternary complexes of the pol I polymerase family obtained in the presence of their dNTP and DNA substrates. In the apo-9 degrees N-7 structure, this conformation appears to be stabilized by an ion pair. Thus far, the other apo-pol alpha structures that have been determined adopt open conformations. These results therefore suggest that the pol alpha polymerases undergo a series of conformational transitions during the catalytic cycle similar to those proposed for the pol I family. Furthermore, comparison of the orientations of the fingers and exonuclease (sub)domains relative to the palm subdomain that contains the pol active site suggests that the exonuclease domain and the fingers subdomain of the polymerase can move as a unit and may do so as part of the catalytic cycle. This provides a possible structural explanation for the interdependence of polymerization and editing exonuclease activities unique to pol alpha family polymerases. We suggest that the NH(2)-terminal domain of 9 degrees N-7 pol may be structurally related to an RNA-binding motif, which appears to be conserved among archaeal polymerases. The presence of such a putative RNA- binding domain suggests a mechanism for the observed autoregulation of bacteriophage T4 DNA polymerase synthesis by binding to its own mRNA. Furthermore, conservation of this domain could indicate that such regulation of pol expression may be a characteristic of archaea. Comparion of the 9 degrees N-7 pol structure to its mesostable homolog from bacteriophage RB69 suggests that thermostability is achieved by shortening loops, forming two disulfide bridges, and increasing electrostatic interactions at subdomain interfaces.


Age-dependent modulation of vascular niches for haematopoietic stem cells.
Blood vessels define local microenvironments in the skeletal system, play crucial roles in osteogenesis and provide niches for haematopoietic stem cells. The properties of niche-forming vessels and their changes in the ageing organism remain incompletely understood. Here we show that Notch signalling in endothelial cells leads to the expansion of haematopoietic stem cell niches in bone, which involves increases in CD31-positive capillaries and platelet-derived growth factor receptor-β (PDGFRβ)-positive perivascular cells, arteriole formation and elevated levels of cellular stem cell factor. Although endothelial hypoxia-inducible factor signalling promotes some of these changes, it fails to enhance vascular niche function because of a lack of arterialization and expansion of PDGFRβ-positive cells. In ageing mice, niche-forming vessels in the skeletal system are strongly reduced but can be restored by activation of endothelial Notch signalling. These findings indicate that vascular niches for haematopoietic stem cells are part of complex, age-dependent microenvironments involving multiple cell populations and vessel subtypes.


The use of biomarkers of toxicity for integrating in vitro hazard estimates into risk assessment for humans.
The role that in vitro systems can play in toxicological risk assessment is determined by the appropriateness of the chosen methods, with respect to the way in which in vitro data can be extrapolated to the in vivo situation. This report presents the results of a workshop aimed at better defining the use of in vitro-derived biomarkers of toxicity (BoT) and determining the place these data can have in human risk assessment. As a result, a conceptual framework is presented for the incorporation of in vitro-derived toxicity data into the risk assessment process. The selection of BoT takes into account that they need to distinguish adverse and adaptive changes in cells. The framework defines the place of in vitro systems in the context of data on exposure, structural and physico-chemical properties, and toxicodynamic and biokinetic modeling. It outlines the determination of a proper point-of-departure (PoD) for in vitro-in vivo extrapolation, allowing implementation in risk assessment procedures. A BoT will need to take into account both the dynamics and the kinetics of the compound in the in vitro systems. For the implementation of the proposed framework it will be necessary to collect and collate data from existing literature and new in vitro test systems, as well as to categorize biomarkers of toxicity and their relation to pathways-of-toxicity. Moreover, data selection and integration need to be driven by their usefulness in a quantitative in vitro-in vivo extrapolation (QIVIVE).


Comprehensive functional genomic resource and integrative model for the human brain.
Despite progress in defining genetic risk for psychiatric disorders, their molecular mechanisms remain elusive. Addressing this, the PsychENCODE Consortium has generated a comprehensive online resource for the adult brain across 1866 individuals. The PsychENCODE resource contains ~79,000 brain-active enhancers, sets of Hi-C linkages, and topologically associating domains; single-cell expression profiles for many cell types; expression quantitative-trait loci (QTLs); and further QTLs associated with chromatin, splicing, and cell-type proportions. Integration shows that varying cell-type proportions largely account for the cross-population variation in expression (with >88% reconstruction accuracy). It also allows building of a gene regulatory network, linking genome-wide association study variants to genes (e.g., 321 for schizophrenia). We embed this network into an interpretable deep-learning model, which improves disease prediction by ~6-fold versus polygenic risk scores and identifies key genes and pathways in psychiatric disorders.


Applied Behavior Analysis, Autism, and Occupational Therapy: A Search for Understanding.
Occupational therapists strive to be mindful, competent practitioners and continuously look for ways to improve practice. Applied behavior analysis (ABA) has strong evidence of effectiveness in helping people with autism achieve goals, yet it does not seem to be implemented in occupational therapy practice. To better understand whether ABA could be an evidence-based option to expand occupational therapy practice, the authors conducted an iterative, multiphase investigation of relevant literature. Findings suggest that occupational therapists apply developmental and sensory approaches to autism treatment. The occupational therapy literature does not reflect any use of ABA despite its strong evidence base. Occupational therapists may currently avoid using ABA principles because of a perception that ABA is not client centered. ABA principles and occupational therapy are compatible, and the two could work synergistically.


Drug export pathway of multidrug exporter AcrB revealed by DARPin inhibitors.
The multidrug exporter AcrB is the inner membrane component of the AcrAB-TolC drug efflux system in Escherichia coli and is responsible for the resistance of this organism to a wide range of drugs. Here we describe the crystal structure of the trimeric AcrB in complex with a designed ankyrin-repeat protein (DARPin) inhibitor at 2.5-A resolution. The three subunits of AcrB are locked in different conformations revealing distinct channels in each subunit. There seems to be remote conformational coupling between the channel access, exit, and the putative proton-translocation site, explaining how the proton motive force is used for drug export. Thus our structure suggests a transport pathway not through the central pore but through the identified channels in the individual subunits, which greatly advances our understanding of the multidrug export mechanism.


Vitamin B12 Enhances Nerve Repair and Improves Functional Recovery After Traumatic Brain Injury by Inhibiting ER Stress-Induced Neuron Injury.
Traumatic brain injury (TBI) is one of the most common causes of neurological damage in young human populations. Vitamin B12 has been reported to promote axon growth of neuronal cells after peripheral nerve injury, which is currently used for the treatment of peripheral nerve damage in the clinical trial. Thus, we hypothesized that TBI can be attenuated by vitaminB12 treatment through its beneficial role on axon regeneration after nerve injury. To confirm it, the biological function of vitaminB12 was characterized using hematoxylin and eosin (H&E) staining, Luxol fast blue (LFB) staining, western blot analysis, and immunohistochemistry staining. The results showed that the neurological functional recovery was improved in the VitaminB12-treated group after TBI, which may be due to downregulation of the endoplasmic reticulum stress-related apoptosis signaling pathway. Moreover, the microtubule stabilization, remyelination and myelin reparation were rescued by vitamin B12, which was consistent with the treatment of 4-phenylbutyric acid (4-PBA), an endoplasmic reticulum stress inhibitor. The study suggests that vitamin B12 may be useful as a novel neuroprotective drug for TBI.


The cis-Pro touch-turn: a rare motif preferred at functional sites.
A new motif of three-dimensional (3D) protein structure is described, called the cis-Pro touch-turn. In this four-residue, three-peptide motif, the central peptide is cis. Residue 2, which precedes the proline, has phi, psi values either in the "prePro region" of the Ramachandran plot near -130 degrees, 75 degrees or in the Lalpha region near +60 degrees, +60 degrees. The Calpha(1)-Calpha(4) distance is 4-5 A and the two flanking peptides lie parallel to one another, making van der Waals contact rather than a hydrogen bond. Apparently, this arrangement is locally unfavorable and therefore rare, usually occurring only if needed for biological function. Of the 12 examples in a 500-protein database, cis-Pro touch-turns are found at the catalytic sites of pectate lyase, Ni-Fe hydrogenase, glucoamylase, xylanase, and opine dehydrogenase and at the primary binding sites of ribonuclease H, type I DNA polymerase, ribotoxin, and phage gene 3 protein. In each of these protein families, the touch-turns serve different roles; their functional importance is supported by conservation and mutagenesis data. In analyzing the conservation patterns of these 3D motifs, new methods for in-depth quality evaluation of the structural bioinformatic data are employed to distinguish between significant exceptions and errors


The impact of gun violence restraining order laws in the U.S. and firearm suicide among older adults: a longitudinal state-level analysis, 2012-2016.
Older adults complete suicide at a disproportionately higher rate compared to the general population, with firearms the most common means of suicide. State gun laws may be a policy remedy. Less is known about Gun Violence Restricting Order (GVRO) laws, which allow for removal of firearms from people deemed to be a danger to themselves or others, and their effects on suicide rates among older adults. The purpose of this study was to examine the association of state firearm laws with the incidence of firearm, non-firearm-related, and total suicide among older adults, with a focus on GVRO laws. This is a longitudinal study of US states using data from 2012 to 2016. The outcome variables were firearm, non-firearm and total suicide rates among older adults. Predictor variables were [1] total number of gun laws to assess for impact of overall firearm legislation at the state level, and [2] GVRO laws. The total number of firearm laws, as well as GVRO laws, were negatively associated with firearm-related suicide rate among older adults ages 55-64 and > 65 years-old (p < 0.001). There was a small but significant positive association of total number of firearm laws to non-firearm-related suicide rates and a negative association with total suicide rate. GVRO laws were not significantly associated with non-firearm-related suicide and were negatively associated with total suicide rate. Stricter firearm legislation, as well as GVRO laws, are protective against firearm-relate suicides among older adults.


The human fatty acid transport protein-1 (SLC27A1; FATP-1) cDNA and gene: organization, chromosomal localization, and expression.
Uptake of fatty acids into cells is a controlled process in part regulated by fatty acid transport proteins (FATPs), which facilitate the transport of fatty acids across the cell membrane. In this study the structure of the human FATP-1 (HGMW-approved symbol SLC27A1) cDNA and gene was determined, and the expression of its mRNA in human was characterized. Muscle and adipose tissue have the highest levels of FATP-1 mRNA, small intestine has intermediate levels, and FATP-1 mRNA is barely detectable in liver. The human FATP-1 gene has 12 exons and extends over more than 13 kb of genomic DNA. The FATP gene maps to chromosome 19p13.1 by fluorescence in situ hybridization, a region previously suggested to be implicated in the determination of small dense low-density lipoprotein (LDL). Knowledge of the gene structure and chromosomal localization will allow screening for FATP mutations in humans with metabolic disorders, whereas knowledge of its expression pattern and factors regulating its expression could be of importance in understanding its biology.


Ocean time-series reveals recurring seasonal patterns of virioplankton dynamics in the northwestern Sargasso Sea.
There are an estimated 10(30) virioplankton in the world oceans, the majority of which are phages (viruses that infect bacteria). Marine phages encompass enormous genetic diversity, affect biogeochemical cycling of elements, and partially control aspects of prokaryotic production and diversity. Despite their importance, there is a paucity of data describing virioplankton distributions over time and depth in oceanic systems. A decade of high-resolution time-series data collected from the upper 300 m in the northwestern Sargasso Sea revealed recurring temporal and vertical patterns of virioplankton abundance in unprecedented detail. An annual virioplankton maximum developed between 60 and 100 m during periods of summer stratification and eroded during winter convective mixing. The timing and vertical positioning of this seasonal pattern was related to variability in water column stability and the dynamics of specific picophytoplankton and heterotrophic bacterioplankton lineages. Between 60 and 100 m, virioplankton abundance was negatively correlated to the dominant heterotrophic bacterioplankton lineage SAR11, as well as the less abundant picophytoplankton, Synechococcus. In contrast, virioplankton abundance was positively correlated to the dominant picophytoplankton lineage Prochlorococcus, and the less abundant alpha-proteobacteria, Rhodobacteraceae. Seasonally, virioplankton abundances were highly synchronous with Prochlorococcus distributions and the virioplankton to Prochlorococcus ratio remained remarkably constant during periods of water column stratification. The data suggest that a significant fraction of viruses in the mid-euphotic zone of the subtropical gyres may be cyanophages and patterns in their abundance are largely determined by Prochlorococcus dynamics in response to water column stability. This high-resolution, decadal survey of virioplankton abundance provides insight into the possible controls of virioplankton dynamics in the open ocean.


Anterior cruciate ligament reconstruction by using otogenous [correction of otogeneous] hamstring tendons with home-based rehabilitation.
We investigated patients undergoing arthroscopic anterior cruciate ligament (ACL) reconstruction using otogenous hamstring tendons with a cross-pin technique to compare a post-operative home-based rehabilitation programme with a clinic-based programme. ACL reconstruction was performed on 104 patients (103 male) by the same surgeon. The mean age of the patients was 31.5 years (range 18 - 43 years) and the mean time interval between injury and operation was 34.3 months. Patients were randomly allocated to either a home-based (n = 52) or clinic-based rehabilitation programme (n = 52). Mean follow-up was 31.1 months (range 12 - 66 months). Patients underwent a series of examinations before and after surgery in order to evaluate functional recovery of their injured knee. The results demonstrated that using otogeneous hamstring tendons for ACL reconstruction was safe and produced satisfactory results. The study also demonstrated that a home-based rehabilitation programme was as effective as a clinic-based programme.


Dynamics of gene silencing during X inactivation using allele-specific RNA-seq.
During early embryonic development, one of the two X chromosomes in mammalian female cells is inactivated to compensate for a potential imbalance in transcript levels with male cells, which contain a single X chromosome. Here, we use mouse female embryonic stem cells (ESCs) with non-random X chromosome inactivation (XCI) and polymorphic X chromosomes to study the dynamics of gene silencing over the inactive X chromosome by high-resolution allele-specific RNA-seq. Induction of XCI by differentiation of female ESCs shows that genes proximal to the X-inactivation center are silenced earlier than distal genes, while lowly expressed genes show faster XCI dynamics than highly expressed genes. The active X chromosome shows a minor but significant increase in gene activity during differentiation, resulting in complete dosage compensation in differentiated cell types. Genes escaping XCI show little or no silencing during early propagation of XCI. Allele-specific RNA-seq of neural progenitor cells generated from the female ESCs identifies three regions distal to the X-inactivation center that escape XCI. These regions, which stably escape during propagation and maintenance of XCI, coincide with topologically associating domains (TADs) as present in the female ESCs. Also, the previously characterized gene clusters escaping XCI in human fibroblasts correlate with TADs. The gene silencing observed during XCI provides further insight in the establishment of the repressive complex formed by the inactive X chromosome. The association of escape regions with TADs, in mouse and human, suggests that TADs are the primary targets during propagation of XCI over the X chromosome.


Significance of oral health in adult patients with congenital heart disease.
Due to improved diagnosis and treatment life expectancy of patients with congenital heart disease is steadily increasing resulting in a growing portion of adult patients. When entering, adulthood patients commonly show a shift in their specific needs for medical care. Since the treatment is mostly not curative many patients have life-long cardiovascular anomalies, among others, entailing high risk for the development of infective endocarditis. Several oral diseases, i.e., caries, apical periodontitis and periodontitis show a very high overall prevalence. These entities are primarily initiated by bacterial infections. Hence, they cause an inherent risk for bacteremia and subsequently for infective endocarditis in patients with congenital heart disease during professional dental care and various daily activities. Conversely congenital heart disease seems to be inevitably associated with considerable impairment of oral health resulting in a tight interrelation between both entities. Different preventive strategies are available to address the elevated risk for infective endocarditis due to oral diseases in patients with congenital heart disease during professional dental care and routine daily activities. This review delineates the current evidence regarding the issue of oral health in adult patients with congenital heart disease.


Utility of long-term video-EEG monitoring for children with staring.
Staring spells are a common reason for referral to overnight epilepsy monitoring unit (EMU) evaluation. However, inpatient EMU admissions are expensive and time consuming. This study determined what percentage of those referred for staring had a confirmed epileptic seizure on long-term video-EEG monitoring (LTM) and developed a scoring system to help prioritize which patients should undergo this procedure. We performed a four-year retrospective chart review of all children at a tertiary pediatric hospital who received LTM (long-term monitoring) for the purposes of characterizing staring. The two goals were to: a) assess how often an LTM admission captured a staring spell that was diagnosed as a seizure and b) determine if any baseline factors predicted this particular positive result. We coded several characteristics of the most recent prior routine EEG if available. We also coded parental reports of the duration, frequency, and breakability of the events as well as post-ictal mental status and the presence/absence of automatisms. Finally, we coded previous neurological and psychiatric diagnoses and medications, as well as family history of epilepsy. Of the 276 admissions, only 29 (11%) captured a staring spell and diagnosed it as seizure. Several baseline variables predicted the likelihood of this positive result. Based on this information, we created a scoring system as follows: -3 points if the previous EEG was normal, -1 point if the child took a medication for a psychiatric condition, +1 point if the child took an anti-epileptic drug for epilepsy, and +1 point if the spells lasted less than 1min. If the total score was zero or less, staring spells diagnosed as seizures rarely occurred (less than 5% of the studies). Our scoring system shows how consideration of prior EEG findings, medication history, and staring spell duration can help prioritize patients for LTM admission to evaluate if staring spells are epileptic seizures.


Anatomy and physiology of the basal ganglia: implications for deep brain stimulation for Parkinson's disease.
Central to surgical management of movement disorders is an understanding of the anatomy and physiology of the basal ganglia. The basal ganglia have been a target for neuromodulation surgery since Russell Meyers' pioneering works in the late 1930s. With the development of deep brain stimulation as the gold standard of surgical intervention for movement disorders, there has been a concomitant evolution in the understanding of the role the basal ganglia plays in the genesis of normal and abnormal motor behaviors. The fundamental concept of the cortico-striato-pallido-thalamocortical loop will be explored in the context of deep brain stimulation. The current targets for deep brain stimulation for Parkinson's disease, the subthalamic nucleus, the globus pallidus internus, and the ventral intermediate nucleus, will be discussed in the framework of the current physiological and anatomical models of Parkinson's disease (PD). Finally, the current understandings of the mechanisms underpinning the beneficial effects of deep brain stimulation for PD will be discussed.


A new method for rapid assignment of S-S bridges in proteins.
A new method for complementing existing protein chemical techniques for the assignment of S-S bridge positions in amino-acid sequences is described. The principle of the method is the direct examination of the masses of protein fragments, obtained by chemical or enzymatic degradation. Proteins are digested under conditions known to minimise disulphide reduction and reshuffling, and the unfractionated digest is examined directly by high field magnet (or other high mass) fast atom bombardment or Californium mass spectrometry. Disulphide linked peptides are identified from their unique masses, and by comparison with the spectrum of digested and reduced samples in which the signal corresponding to the S-S linked peptide(s) is replaced by two signals corresponding to the respective thiol peptide components, if INTER-bridged, or shifted by two mass units (dithiol) if INTRA-bridged. This rapid procedure has considerable potential in assisting with studies on the primary structure of proteins, in crystallographic studies and the monitoring of denaturation/renaturation of recombinant proteins.


Digital imaging and radiographic practise in diagnostic radiography: An overview of current knowledge and practice in Europe.
Recent research has identified the issue of 'dose creep' in diagnostic radiography and claims it is due to the introduction of CR and DR technology. More recently radiographers have reported that they do not regularly manipulate exposure factors for different sized patients and rely on pre-set exposures. The aim of the study was to identify any variation in knowledge and radiographic practice across Europe when imaging the chest, abdomen and pelvis using digital imaging. A random selection of 50% of educational institutes (n = 17) which were affiliated members of the European Federation of Radiographer Societies (EFRS) were contacted via their contact details supplied on the EFRS website. Each of these institutes identified appropriate radiographic staff in their clinical network to complete an online survey via SurveyMonkey. Data was collected on exposures used for 3 common x-ray examinations using CR/DR, range of equipment in use, staff educational training and awareness of DRL. Descriptive statistics were performed with the aid of Excel and SPSS version 21. A response rate of 70% was achieved from the affiliated educational members of EFRS and a rate of 55% from the individual hospitals in 12 countries across Europe. Variation was identified in practice when imaging the chest, abdomen and pelvis using both CR and DR digital systems. There is wide variation in radiographer training/education across countries. There is a need for standardisation of education and training including protocols and exposure parameters to ensure that there is continued adherence to the ALARA principle.


The effect of spacing regularity on visual crowding.
Crowding limits peripheral visual discrimination and recognition: a target easily identified in isolation becomes impossible to recognize when surrounded by other stimuli, often called flankers. Most accounts of crowding predict less crowding when the target-flanker distance increases. On the other hand, the importance of perceptual organization and target-flanker coherence in crowding has recently received more attention. We investigated the effect of target-flanker spacing on crowding in multi-element stimulus arrays. We show that increasing the average distance between the target and the flankers does not always decrease the amount of crowding but can even sometimes increase it. We suggest that the regularity of inter-element spacing plays an important role in determining the strength of crowding: regular spacing leads to the perception of a single, coherent, texture-like stimulus, making judgments about the individual elements difficult.


Extrapyramidal Symptoms Probably Related to Risperidone Treatment: A Case Series.
Atypical antipsychotics, like risperidone purportedly, score over their typical counterparts in terms of their lower propensity toward producing extrapyramidal symptoms (EPS). However, recent studies have furnished evidence to the contrary. Hereby, we present a case series implicating risperidone as the causative agent for EPS. As a part of the pharmacovigilance programme of India, the authors have assessed 10 physician-reported cases of EPS among the 1,830 patients who were prescribed risperidone within the time period of January 2012-December 2014 in a tertiary care hospital in South India. Causality, severity, and preventability assessments of adverse reaction were done as per Naranjo's, Hartwig's, and Thornton'scale respectively. Of the 10 cases, a dose-dependent occurrence of EPS was noted in all and the time duration for development of EPS ranged from 1 week to 2 years. Four patients developed EPS at a dose of 6-8 mg, 4 developed at a dose of 4-6 mg, and the remaining 2 developed at 2 and 1 mg. A strong temporal correlation between risperidone and EPS was noted in all cases. High doses produced EPS early, whereas moderate to low doses produced EPS at a later date. Thus, cautious use and close monitoring are warranted in the chronic use of risperidone.


Age alone is not a risk factor for left atrial thrombus in atrial fibrillation.
Advanced age is an indication for anticoagulation in patients with atrial fibrillation though it is not clear that elderly patients have a higher prevalence of left atrial thrombus. The purpose of this study was to clarify whether advanced age represents a risk for left atrial thrombus formation irrespective of other clinical variables. Observational study in patients with atrial fibrillation undergoing a transoesophageal echo scan for various clinical indications. University-affiliated cardiology service in a general hospital. Results are reported in 381 patients, 257 aged less than 75 years (Gr. A) and 124 aged 75 years or more (Gr. B). Thrombi were detected by TOE in 30 patients (7.9%), 21 from group A and nine from group B (8.1% vs 7.2%, p = NS). No thrombi were detected in patients with lone atrial fibrillation. Among patients with either valvular or nonvalvular atrial fibrillation, left atrial thrombus presence was not related to age or anticoagulation status. In patients with atrial fibrillation, age itself does not predict the presence of left atrial thrombus and the only identifiable risk factor seems to be the existing cardiac pathology.


Characterization of "Hydrocarbon" Dry Cleaning in King County, Washington.
In King County, Washington, the most frequently used alternative solvent to perchloroethylene is a hydrotreated petroleum hydrocarbon. The objectives of the authors' study were to 1) determine the frequency of use of process chemicals used in "hydrocarbon" dry cleaning and gather other operational information; 2) chemically characterize the process chemicals; 3) characterize the still bottoms and separator water wastes according to dangerous waste and wastewater discharge regulations; 4) identify linkages between work practices, process chemicals, and the chemical composition of the waste streams; and 5) evaluate the aquatic toxicity of the hydrocarbon solvent and detergent. Many hydrocarbon dry cleaners are using process chemicals that contain hazardous substances, including trichloroethylene. One sample of separator water contained 13,000 µg/L trichloroethylene. This sample was determined to be federal hazardous waste, state-only dangerous waste (i.e., according to Washington state-specific regulations), and failed wastewater discharge thresholds. All still bottoms were determined to be state-only dangerous wastes. Efforts should be directed towards replacing hazardous spot cleaning chemicals with safer alternatives and ensuring that wastes are disposed of appropriately.


Polarization-based non-staining cell detection.
Polarization is an important characteristic of electromagnetic waves, which can not be detected by either the human visual system or traditional image sensors. Motivated by various animal species with polarization vision as well as by the prospect of improving the image quality of the imaging systems, we are exploring the potential of polarization for microscope imaging. The most powerful techniques for molecule monitoring requires complex preprocessing for labeling the sample with different dyes. In this paper, we propose a cell detection method using polarization imaging without any need for staining target cell samples with any chemical dye. The motivation for this work is to develop an optical imaging technique that is simple and that can be used on live cells. The polarization sensitivity of cell samples is studied in this paper. A definition for the quantity called "polarization deviation" is proposed in order to identify clearer the difference between target cells and the background. Based on the polarization deviation detection method, a three-parameter polarization imaging method is employed to further simplify the image capture procedure for the proposed label-free cell detection. A color imaging methodology based on the well-known color space is utilized in order to represent the captured polarization information using computer graphics.


Laparoscopic-assisted renal autotransplantation.
To report our experience with laparoscopic nephrectomy and autotransplantation for the management of a variety of conditions with significant loss of healthy ureteral tissue or ureteral length. Renal autotransplantation has been described as an effective method for addressing this problem, avoiding the need for nephrectomy or complex ureteral replacement. In an effort to decrease the morbidity associated with traditional autotransplantation we elected to perform laparoscopic procurement of the kidney. Four patients underwent laparoscopic nephrectomy using a transperitoneal four-port technique and subsequent autotransplantation into the iliac fossa for the treatment of proximal ureteral avulsion (2 patients), ureteral malignancy, and ureteral stricture. All patients had less than 5 cm of viable ureter. All procedures were performed without intraoperative complications. All renal scans on postoperative day 1 demonstrated good perfusion. None of the patients had a postoperative rise in serum creatinine. On postoperative day 1, the mean creatinine value was 0.95 mg/dL. Three patients had an uneventful postoperative course. One patient, however, had loss of the graft because of renal vein thrombosis on postoperative day 7. She was later found to have an undiagnosed thrombophilic disorder (decreased levels of antithrombin III) and to have a recent history of oral contraceptive use. Laparoscopic nephrectomy with renal autotransplantation is a feasible minimally invasive alternative to treat patients who have significant ureteral loss. This approach avoids the need for an upper abdominal or flank incision, resulting in decreased morbidity. The initial follow-up studies indicated stable renal function. Additional long-term observation is currently under way.


Determining the effectiveness of High Resolution Melting analysis for SNP genotyping and mutation scanning at the TP53 locus.
Together single nucleotide substitutions and small insertion/deletion variants are the most common form of sequence variation in the human gene pool.High-resolution SNP profile and/or haplotype analyses enable the identification of modest-risk susceptibility genes to common diseases, genes that may modulate responses to pharmaceutical agents, and SNPs that can affect either their expression or function. In addition, sensitive techniques for germline or somatic mutation detection are important tools for characterizing sequence variations in genes responsible for tumor predisposition. Cost-effective methods are highly desirable. Many of the recently developed high-throughput technologies are geared toward industrial scale genetic studies and arguably do not provide useful solutions for small laboratory investigator-initiated projects. Recently, the use of new fluorescent dyes allowed the high-resolution analysis of DNA melting curves (HRM). Here, we compared the capacity of HRM, applicable to both genotyping and mutation scanning, to detect genetic variations in the tumor suppressor gene TP53 with that of mutation screening by full resequencing. We also assessed the performance of a variety of available HRM-based genotyping assays by genotyping 30 TP53 SNPs. We describe a series of solutions to handle the difficulties that may arise in large-scale application of HRM to mutation screening and genotyping at the TP53 locus. In particular, we developed specific HRM assays that render possible genotyping of 2 or more, sometimes closely spaced, polymorphisms within the same amplicon. We also show that simultaneous genotyping of 2 SNPs from 2 different amplicons using a multiplex PCR reaction is feasible; the data can be analyzed in a single HRM run, potentially improving the efficiency of HRM genotyping workflows. The HRM technique showed high sensitivity and specificity (1.0, and 0.8, respectively, for amplicons of <400 bp) for mutation screening and provided useful genotyping assays as assessed by comparing the results with those obtained with Sanger sequencing. Thus, HRM is particularly suitable for either performing mutation scanning of a large number of samples, even in the situation where the amplicon(s) of interest harbor a common variant that may disturb the analysis, or in a context where gathering common SNP genotypes is of interest.


Efficacy of azithromycin in the treatment of childhood typhoid Fever.
An intervention study was carried out in Paediatric wards for a period of one year from January 2003 to December 2003 to determine the efficacy and safety of azithromycin in the treatment of uncomplicated childhood typhoid fever. A total of 50 cases were enrolled in the study. The inclusion criteria of the cases were: documented fever for more than 7 days plus two or more of the following clinical features: toxic appearance, abdominal tenderness, hepatomegaly, splenomegaly, diarrhoea, constipation and coated tongue plus positive Widal test and/or blood culture positivity. Patients who had complication like gastrointestinal tract (GIT) haemorrhage; intestinal perforation and/or shock were excluded from the study. Data were collected in a structured questionnaire. Azithromycin was given at a dose of 10mg/kg /day for a period of 07 days. The time to defervescence was 3.82+/-1.49 days. The minimum defervescence time was 02 days and maximum was 07 days. Clinical cure rate was 94%. No serious adverse effect was noted related to azithromycin therapy except nausea, vomiting, and jaundice. Prior treatment with antibiotics did not affect defervescence time (P>0.05). Pre-treatment febrile period has got positive and linear correlation with clinical response (r = +0.593). It was found that once daily administration of oral azithromycin for seven days in the treatment of uncomplicated typhoid fever was effective and reasonably safe.


A Randomized Controlled Trial of the Shikamana Intervention to Promote Antiretroviral Therapy Adherence Among Gay, Bisexual, and Other Men Who Have Sex with Men in Kenya: Feasibility, Acceptability, Safety and Initial Effect Size.
Gay, bisexual, and other men who have sex with men (GBMSM) living with HIV in rights-constrained settings need support for antiretroviral therapy (ART) adherence due to barriers including stigma. The Shikamana intervention combined modified Next Step Counseling by providers with support from trained peers to improve adherence among GBMSM living with HIV in Kenya. A randomized controlled trial with 6-month follow-up was used to determine feasibility, acceptability, safety, and initial intervention effects. Generalized estimating equations examined differences in self-reported adherence and virologic suppression. Sixty men enrolled, with 27 randomly assigned to the intervention and 33 to standard care. Retention did not differ by arm, and no adverse events occurred. Feedback on feasibility and acceptability was positive based on exit interviews. After adjustment for baseline viral suppression and confounding, the intervention group had a sixfold increased odds of viral suppression during follow-up. A larger trial of a scaled-up intervention is needed.


Fibre based cellular transfection.
Optically assisted transfection is emerging as a powerful and versatile method for the delivery of foreign therapeutic agents to cells at will. In particular the use of ultrashort pulse lasers has proved an important route to transiently permeating the cell membrane through a multiphoton process. Though optical transfection has been gaining wider usage to date, all incarnations of this technique have employed free space light beams. In this paper we demonstrate the first system to use fibre delivery for the optical transfection of cells. We engineer a standard optical fibre to generate an axicon tip with an enhanced intensity of the remote output field that delivers ultrashort (~ 800 fs) pulses without requiring the fibre to be placed in very close proximity to the cell sample. A theoretical model is also developed in order to predict the light propagation from axicon tipped and bare fibres, in both air and water environments. The model proves to be in good agreement with the experimental findings and can be used to establish the optimum fibre parameters for successful cellular transfection. We readily obtain efficiencies of up to 57 % which are comparable with free space transfection. This advance paves the way for optical transfection of tissue samples and endoscopic embodiments of this technique.


Spatial Coherence of Spin-Orbit-Coupled Bose Gases.
Spin-orbit-coupled Bose-Einstein condensates (SOBECs) exhibit two new phases of matter, now known as the stripe and plane-wave phases. When two interacting spin components of a SOBEC spatially overlap, density modulations with periodicity given by the spin-orbit coupling strength appear. In equilibrium, these components fully overlap in the miscible stripe phase and overlap only in a domain wall in the immiscible plane-wave phase. Here we probe the density modulation present in any overlapping region with optical Bragg scattering and observe the sudden drop of Bragg scattering as the overlapping region shrinks. Using an atomic analog of the Talbot effect, we demonstrate the existence of long-range coherence between the different spin components in the stripe phase and surprisingly even in the phase-separated plane-wave phase.


Comparison between Source-induced Dissociation and Collision-induced Dissociation of Ampicillin, Chloramphenicol, Ciprofloxacin, and Oxytetracycline via Mass Spectrometry.
Mass spectrometry (MS) is a very powerful instrument that can be used to analyze a wide range of materials such as proteins, peptides, DNA, drugs, and polymers. The process typically involves either chemical or electron (impact) ionization of the analyte. The resulting charged species or fragment is subsequently identified by the detector. Usually, single mass uses source-induced dissociation (SID), whereas mass/mass uses collision-induced dissociation (CID) to analyze the chemical fragmentations Each technique has its own advantages and disadvantages. While CID is most effective for the analysis of pure substances, multiple- step MS is a powerful technique to get structural data. Analysis of veterinary drugs ampicillin, chloramphenicol, ciprofloxacin, and oxytetracycline serves to highlight the slight differences between SID and CID. For example, minor differences were observed between ciprofloxacin and oxytetracycline via SID or CID. However, distinct fragmentation patterns were observed for ampicllin depending on the analysis method. Both SID and CID showed similar fragmentation spectra but different signal intensities for chloramphenicol. There are several factors that can influence the fragmentation spectra, such as the collision energy, major precursor ion, electrospray mode (positive or negative), and sample homogeneity. Therefore, one must select a fragmentation method on an empirical and case-by-case basis.


Utility of an Alzheimer's Disease Risk-Weighted Polygenic Risk Score for Predicting Rates of Cognitive Decline in Preclinical Alzheimer's Disease: A Prospective Longitudinal Study.
With the exception of APOE, genetic variants associated with increased Alzheimer's disease (AD) risk are characterized by small effect sizes. Polygenic risk scores (PRS) have shown utility in predicting AD risk; however, their utility for predicting decline in cognition at preclinical stages of AD is poorly understood. To validate associations of a 22-variant AD-risk-weighted PRS with AD risk and related biomarkers and to assess its utility to predict cognitive decline. The PRS was evaluated with respect to brain amyloid-β (Aβ) burden, cerebrospinal fluid (CSF) Aβ42, total-tau, and phospho-tau, and decline in cognition in 643 (570 cognitively normal (CN), 73 AD) PET-imaged participants from the longitudinal Australian Imaging, Biomarkers and Lifestyle (AIBL) Study of Ageing. Cognition was assessed using three composite measures; global cognition, verbal episodic memory, and a Pre-Alzheimer's Cognitive Composite (PACC). PRS, both with and without APOE, were positively correlated with brain Aβ burden, CSF total-tau, and phospho-tau in CN older adults. Further, in CN biomarker positive (Aβhigh) participants, significant associations were observed with baseline and longitudinal cognition. However, this association was not observed after the removal of APOE. Partitioning the PRS into quartiles revealed that the PRS associations with cognitive decline in Aβhigh CN older adults is due to a saturating effect of APOE genotype. An AD-risk-weighted PRS is associated with cognitive decline in CN older adults. However, this association is absent when APOE genotype is excluded from the PRS, suggesting that associations with cognitive decline in this model of polygenic risk are driven by APOE genotype alone. Further research is needed to define appropriate PRSs with greater utility for predicting preclinical AD cognitive decline.


Differential binding of serum glycoproteins to lectins during hepatic regeneration in hepatocellular carcinoma and fulminant hepatic failure.
1. The concentrations of four serum glycoproteins, thyroxine-binding globulin, alpha 2-macroglobulin, alpha 1-antitrypsin and transferrin, as well as their reactivities with concanavalin A and lentil-lectin, were measured in patients with hepatocellular carcinoma or fulminant hepatic failure and in normal subjects. 2. Serum concentrations of thyroxine-binding globulin and alpha 1-antitrypsin were significantly greater in patients with hepatocellular carcinoma than in normal subjects, and the percentage lentil-lectin reactivity of these two proteins was markedly increased. 3. With the exception of transferrin, which did not bind to lentil-lectin, an enhancement of lentil-lectin reactivity was observed for the glycoproteins in serum from patients with fulminant hepatic failure. No difference in concanavalin A binding was found between the groups for any of the glycoproteins. 4. Altered fucosylation, as indicated by increased lentil-lectin binding, occurs in several glycoproteins arising in malignant and non-malignant conditions associated with abnormal hepatic regeneration.


Somatostatin and its receptor family.
Somatostatin (SST), a regulatory peptide, is produced by neuroendocrine, inflammatory, and immune cells in response to ions, nutrients, neuropeptides, neurotransmitters, thyroid and steroid hormones, growth factors, and cytokines. The peptide is released in large amounts from storage pools of secretory cells, or in small amounts from activated immune and inflammatory cells, and acts as an endogenous inhibitory regulator of the secretory and proliferative responses of target cells that are widely distributed in the brain and periphery. These actions are mediated by a family of seven transmembrane (TM) domain G-protein-coupled receptors that comprise five distinct subtypes (termed SSTR1-5) that are endoded by separate genes segregated on different chromosomes. The five receptor subtypes bind the natural SST peptides, SST-14 and SST-28, with low nanomolar affinity. Short synthetic octapeptide and hexapeptide analogs bind well to only three of the subtypes, 2, 3, and 5. Selective nonpeptide agonists with nanomolar affinity have been developed for four of the subtypes (SSTR1, 2, 3, and 4) and putative peptide antagonists for SSTR2 and SSTR5 have been identified. The ligand binding domain for SST ligands is made up of residues in TMs III-VII with a potential contribution by the second extracellular loop. SSTRs are widely expressed in many tissues, frequently as multiple subtypes that coexist in the same cell. The five receptors share common signaling pathways such as the inhibition of adenylyl cyclase, activation of phosphotyrosine phosphatase (PTP), and modulation of mitogen-activated protein kinase (MAPK) through G-protein-dependent mechanisms. Some of the subtypes are also coupled to inward rectifying K(+) channels (SSTR2, 3, 4, 5), to voltage-dependent Ca(2+) channels (SSTR1, 2), a Na(+)/H(+) exchanger (SSTR1), AMPA/kainate glutamate channels (SSTR1, 2), phospholipase C (SSTR2, 5), and phospholipase A(2) (SSTR4). SSTRs block cell secretion by inhibiting intracellular cAMP and Ca(2+) and by a receptor-linked distal effect on exocytosis. Four of the receptors (SSTR1, 2, 4, and 5) induce cell cycle arrest via PTP-dependent modulation of MAPK, associated with induction of the retinoblastoma tumor suppressor protein and p21. In contrast, SSTR3 uniquely triggers PTP-dependent apoptosis accompanied by activation of p53 and the pro-apoptotic protein Bax. SSTR1, 2, 3, and 5 display acute desensitization of adenylyl cyclase coupling. Four of the subtypes (SSTR2, 3, 4, and 5) undergo rapid agonist-dependent endocytosis. SSTR1 fails to be internalized but is instead upregulated at the membrane in response to continued agonist exposure. Among the wide spectrum of SST effects, several biological responses have been identified that display absolute or relative subtype selectivity. These include GH secretion (SSTR2 and 5), insulin secretion (SSTR5), glucagon secretion (SSTR2), and immune responses (SSTR2).


Diabetic endothelial dysfunction: the role of poly(ADP-ribose) polymerase activation.
Diabetic patients frequently suffer from retinopathy, nephropathy, neuropathy and accelerated atherosclerosis. The loss of endothelial function precedes these vascular alterations. Here we report that activation of poly(ADP-ribose) polymerase (PARP) is an important factor in the pathogenesis of endothelial dysfunction in diabetes. Destruction of islet cells with streptozotocin in mice induced hyperglycemia, intravascular oxidant production, DNA strand breakage, PARP activation and a selective loss of endothelium-dependent vasodilation. Treatment with a novel potent PARP inhibitor, starting after the time of islet destruction, maintained normal vascular responsiveness, despite the persistence of severe hyperglycemia. Endothelial cells incubated in high glucose exhibited production of reactive nitrogen and oxygen species, consequent single-strand DNA breakage, PARP activation and associated metabolic and functional impairment. Basal and high-glucose-induced nuclear factor-kappaB activation were suppressed in the PARP-deficient cells. Our results indicate that PARP may be a novel drug target for the therapy of diabetic endothelial dysfunction.


Reciprocal obligations for prevention of occupationally acquired tuberculosis among healthcare workers.
As I began my work on occupationally acquired tuberculosis (TB), I was perturbed by a series of media reports on TB among healthcare workers (HCWs) in India. This included a report on the death of a resident doctor who was suffering from multidrug-resistant (MDR) TB. The risk of occupationally acquired TB is well documented. A few studies have reported an increased risk of TB among HCWs in developing countries, including India.


The relation of patient satisfaction with treatment of otitis externa to clinical outcomes: development of an instrument.
This survey was undertaken to develop a short, comprehensive measure of patient satisfaction with pharmacologic treatment for otitis externa and to assess the relationships between satisfaction, disease symptoms, and medication side effects. Otitis externa was diagnosed in 41 patients recruited from 6 sites; 34 patients completed and returned the study instruments and were included in the study. Patients or their caregivers administered polymyxin/neomycin/hydrocortisone ear drops prescribed by a physician and completed a daily diary for 10 days and a satisfaction questionnaire at the end of the treatment period. The main outcome measures were the subscale scores for patient satisfaction and their relation to medication side effects, symptoms of ear infection, activity limitations, pain, and adherence to prescription regimens. The questionnaire and its subscales demonstrated good psychometric properties (ie, reliability coefficients >0.75, except for 1 subscale). Overall satisfaction was found to be significantly correlated with relief of symptoms, ability to return to normal activities, ease of administration, and medication side effects. Satisfaction subscale scores were correlated with patient-reported severity of medication side effects and disease symptoms. More than half the patients took drops for more than the prescribed number of days, and one third took more than the prescribed number of drops per administration (ie, overadherence). The relation between satisfaction and adherence was weak, perhaps due to the high rates of overadherence. Our results demonstrate that patient satisfaction with otic medication can be assessed across various aspects of satisfaction and that it is correlated with reported disease symptoms and medication side effects. This type of multifaceted assessment may help physicians select between medications with different side-effect profiles and administration schedules. Larger studies are needed to evaluate the relationship between satisfaction with an otic medication and adherence to a medication regimen.


Effects of aging on environmental modulation of hippocampal evoked responses.
Evoked responses in the dentate gyrus of the hippocampal formation undergo a long-term enhancement following high-frequency stimulation of the perforant pathway. A similar change results from exposure of animals to a complex spatial environment. The effect of aging on the development and decay of this environmentally induced response enhancement was examined in the present study. Previously it was shown that electrically induced enhancement reaches the same asymptotic level in young and old animals but decays more quickly in old animals. It has been suggested that this faster decay may underlie the faster forgetting of spatial information observed in old animals. Chronic recordings were made from young (14 month) and old (32 month) rats. After exposure to an enriched environment for 11 days, the population spike component of the response increased about 125% over baseline in both groups. No changes were seen in either group in the synaptic component. Following the enrichment treatment, animals were returned to their home cages. The decay of the enhanced population spike during this period differed markedly between age-groups (time constants of 30 and 11 days for the young and old groups, respectively). These results suggest that the factors governing the decay of electrically and environmentally induced response enhancement are similarly affected by the aging process and may share a common mechanism.


The role of cis-acting promoter elements in tissue-specific albumin gene expression.
The mouse albumin gene promoter has six closely spaced binding sites for nuclear proteins that are located between the TATA motif and nucleotide position -170. In vitro transcription with liver or spleen nuclear extracts of templates containing either mutated or polymerized albumin promoter elements establishes a hierarchy of the different protein binding sites for tissue-specific albumin gene transcription. The HNF-1 and C/EBP binding sites strongly activate transcription in a tissue-specific manner. The NF-Y binding site has a lower activation potential and is less specific, being equally efficient in liver and spleen nuclear extracts. The remaining elements are relatively weak activator sites.


Effect of post-annealing on the plasma etching of graphene-coated-copper.
High temperature deposition of graphene on Cu by chemical vapor deposition can be used to produce high quality films. However, these films tend to have a non-equilibrium structure, with relatively low graphene adhesion. In this study, samples of graphene grown on copper foils by high temperature CVD were post-deposition annealed at temperatures well below the critical temperature of Cu. Resistance to etching under plasma was examined to assess the mechanical robustness of the graphene on the Cu surface, analyzed using optical and Raman microscopies. We found a correlation between the post-annealing time and etching time for the complete removal of graphene from Cu. Etching rates, minimum etch times, and surface appearance were observed to vary depending on the etching plasma (air, oxygen or nitrogen). Oxygen plasmas were found to be the least aggressive, emphasizing the improved adhesion with post-annealing treatments. Our results imply that the etching of graphene on Cu, and hence the adhesion of graphene, can be controlled by proper annealing and choice of plasma gas.


What is dopamine doing in model-based reinforcement learning?
Experiments have implicated dopamine in model-based reinforcement learning (RL). These findings are unexpected as dopamine is thought to encode a reward prediction error (RPE), which is the key teaching signal in model-free RL. Here we examine two possible accounts for dopamine's involvement in model-based RL: the first that dopamine neurons carry a prediction error used to update a type of predictive state representation called a successor representation, the second that two well established aspects of dopaminergic activity, RPEs and surprise signals, can together explain dopamine's involvement in model-based RL.


Characterization of a Pipecolic Acid Biosynthesis Pathway Required for Systemic Acquired Resistance.
Systemic acquired resistance (SAR) is an immune response induced in the distal parts of plants following defense activation in local tissue. Pipecolic acid (Pip) accumulation orchestrates SAR and local resistance responses. Here, we report the identification and characterization of SAR-DEFICIENT4 (SARD4), which encodes a critical enzyme for Pip biosynthesis in Arabidopsis thaliana Loss of function of SARD4 leads to reduced Pip levels and accumulation of a Pip precursor, Δ1-piperideine-2-carboxylic acid (P2C). In Escherichia coli, expression of the aminotransferase ALD1 leads to production of P2C and addition of SARD4 results in Pip production, suggesting that a Pip biosynthesis pathway can be reconstituted in bacteria by coexpression of ALD1 and SARD4. In vitro experiments showed that ALD1 can use l-lysine as a substrate to produce P2C and P2C is converted to Pip by SARD4. Analysis of sard4 mutant plants showed that SARD4 is required for SAR as well as enhanced pathogen resistance conditioned by overexpression of the SAR regulator FLAVIN-DEPENDENT MONOOXYGENASE1. Compared with the wild type, pathogen-induced Pip accumulation is only modestly reduced in the local tissue of sard4 mutant plants, but it is below detection in distal leaves, suggesting that Pip is synthesized in systemic tissue by SARD4-mediated reduction of P2C and biosynthesis of Pip in systemic tissue contributes to SAR establishment.


Evidence that nuclear factor IA inhibits repair after white matter injury.
Chronic demyelination can result in axonopathy and is associated with human neurological conditions such as multiple sclerosis (MS) in adults and cerebral palsy in infants. In these disorders, myelin regeneration is inhibited by impaired differentiation of oligodendrocyte progenitors into myelin-producing oligodendrocytes. However, regulatory factors relevant in human myelin disorders and in myelin regeneration remain poorly understood. Here we have investigated the role of the transcription factor nuclear factor IA (NFIA) in oligodendrocyte progenitor differentiation during developmental and regenerative myelination. NFIA expression patterns in human neonatal hypoxic-ischemic encephalopathy (HIE) and MS as well as developmental expression in mice were evaluated. Functional studies during remyelination were performed using a lysolecithin model, coupled with lentiviral misexpression of NFIA. The role of NFIA during oligodendrocyte lineage development was characterized using chick and mouse models and in vitro culture of oligodendrocyte progenitors. Biochemical mechanism of NFIA function was evaluated using chromatin immunoprecipitation and reporter assays. NFIA is expressed in oligodendrocyte progenitors, but not differentiated oligodendrocytes during mouse embryonic development. Examination of NFIA expression in white matter lesions of human newborns with neonatal HIE, as well active MS lesions in adults, revealed that it is similarly expressed in oligodendrocyte progenitors and not oligodendrocytes. Functional studies indicate that NFIA is sufficient to suppress oligodendrocyte progenitor differentiation during adult remyelination and embryonic development through direct repression of myelin gene expression. These studies suggest that NFIA participates in the control of oligodendrocyte progenitor differentiation and may contribute to the inhibition of remyelination in human myelin disorders.


Computer-based reading and spelling practice for children with learning disabilities.
To examine the effects of computer-based reading and spelling practice on the development of reading and spelling skills, a pretest-training-posttest experiment was conducted in The Netherlands. Eleven girls and 17 boys with written language disorders (on the average, 9 years, 7 months old and performing 2 grades below age expectancy) practiced hard-to-read words under three conditions: reading from the computer screen, copying from the screen, and writing from memory after presentation on the screen. For all words, whole-word sound was available on call during practice. To assess learning effects, both a dictation and a read-aloud task were administered in which nonpracticed control words were also presented. During training, the computer kept record of several aspects of the pupils' learning behavior. It was found that copying words from the screen resulted in significantly fewer spelling errors on the posttest than writing words from memory, and that both forms of spelling practice led to fewer spelling errors than only reading words during practice. All three forms of practice improved to the same degree both the accuracy and fluency of reading the practiced words aloud. The way in which spelling and reading practice, in combination with speech feedback, support the development of phonological skills in children with written language disorders is highlighted in the discussion.


Long-distance migrants vary migratory behaviour as much as short-distance migrants: An individual-level comparison from a seabird species with diverse migration strategies.
As environmental conditions fluctuate across years, seasonal migrants must determine where and when to move without comprehensive knowledge of conditions beyond their current location. Animals can address this challenge by following cues in their local environment to vary behaviour in response to current conditions, or by moving based on learned or inherited experience of past conditions resulting in fixed behaviour across years. It is often claimed that long-distance migrants are more fixed in their migratory behaviour because as distance between breeding and wintering areas increases, reliability of cues to predict distant and future conditions decreases. While supported by some population-level studies, the influence of migration distance on behavioural variation is seldom examined on an individual level. Lesser black-backed gulls Larus fuscus are generalist seabirds that use a diversity of migration strategies. Using high-resolution multi-year GPS tracking data from 82 individuals from eight colonies in Western Europe, we quantified inter- and intra-individual variation in non-breeding distributions, winter site fidelity, migration routes and timing of migration, with the objectives of determining how much variation lesser black-backed gulls have in their migratory behaviour and examining whether variation changes with migration distance. We found that intra-individual variation was significantly lower than variation between individuals for non-breeding distributions, winter site fidelity, migration routes and timing of migration, resulting in consistent individual strategies for all behaviours examined. Yet, intra-individual variation ranged widely among individuals (e.g. winter site overlap: 0-0.91 out of 1; migration timing: 0-192 days), and importantly, individual differences in variation were not related to migration distance. The apparent preference for maintaining a consistent strategy, present in even the shortest distance migrants, suggests that familiarity may be more advantageous than exactly tracking current environmental conditions. Yet, variation in behaviour across years was observed in many individuals and could be substantial. This suggests that individuals, irrespective of migration distance, have the capacity to adjust to current conditions within the broad confines of their individual strategies, and occasionally, even change their strategy.


Effect of age on hypothalamic prostaglandin E2 production and fever in response to tumour necrosis factor (cachectin) and endotoxin in rats.
1. Decreased febrile responses to interleukin-1 and endotoxin have been noted in a number of species with ageing. 2. The present study extends these observations by examining the pyrogenic response to intravenous recombinant human tumour necrosis factor-alpha (50 micrograms/kg) using conscious rats aged 7,20 and 80 weeks. 3. The febrile response decreased in magnitude and duration with age. Fevers of 0.9 degree C and of 5 h duration were observed in the youngest rats, whereas those aged 80 weeks were afebrile. The depression in serum zinc level and the elevation in liver zinc level, which occurred 7 h after injection, were unaffected by age. 4. The mechanism of the reduced pyrogenic response was examined by assessing prostaglandin E2 production in vitro from hypothalami of rats, aged 10 and 24 weeks, in response to Escherichia coli endotoxin and tumour necrosis factor. 5. Whereas the production of prostaglandin E2 increased by 47% and 52%, respectively, in hypothalami from 10-week-old rats, no response to either pyrogen was obtained in tissue from rats aged 24 weeks. 6. Maturity brings about a decreased responsiveness of hypothalamic prostaglandin E2 production to pyrogens, which may explain the decreased febrile responses observed.


Genetic variation in the lymphotoxin-α (LTA)/tumour necrosis factor-α (TNFα) locus as a risk factor for idiopathic achalasia.
Idiopathic achalasia is a rare motor disorder of the oesophagus characterised by neuronal loss at the lower oesophageal sphincter. Achalasia is generally accepted as a multifactorial disorder with various genetic and environmental factors being risk-associated. Since genetic factors predisposing to achalasia have been poorly documented, we assessed whether single nucleotide polymorphisms (SNPs) in genes mediating immune response and neuronal function contribute to achalasia susceptibility. 391 SNPs covering 190 immune and 67 neuronal genes were genotyped in an exploratory cohort from Central Europe (589 achalasia patients, 794 healthy volunteers (HVs)). 24 SNPs (p<0.05) were validated in an Italian (160 achalasia patients, 278 HVs) and Spanish cohort (281 achalasia patients, 296 HVs). 16 SNPs in linkage disequilibrium (LD) with rs1799724 (r(2)>0.2) were genotyped in the exploratory cohort. Genotype distributions of patients (1030) and HVs (1368) were compared using Cochran-Armitage trend test. The rs1799724 SNP located between the lymphotoxin-α (LTA) and tumour necrosis factor-α (TNFα) genes was significantly associated with achalasia and withstood correction for testing multiple SNPs (p=1.17E-4, OR=1.41 (1.18 to 1.67)). SNPs in high LD with rs1799724 were associated with achalasia. Three SNPs located in myosin-5B, adrenergic receptor-β-2 and interleukin-13 (IL13) showed nominally significant association to achalasia that was strengthened by replication. Our study provides evidence for rs1799724 at the LTA/TNFα locus as a susceptibility factor for idiopathic achalasia. Additional studies are needed to dissect which genetic variants in the LTA/TNFα locus are disease-causing and confirm other variants as potential susceptibility factors for achalasia.


Personality and problem gambling: a prospective study of a birth cohort of young adults.
Individual differences in dimensions of personality may play an important role in explaining risk for disordered gambling behavior as well as the comorbidity between disordered gambling behavior and other substance-related addictive disorders. To identify the personality correlates of problem gambling in a representative non-treatment-seeking sample, as well as to determine whether these are similar to the personality correlates of other substance-related addictive disorders and whether individual differences in personality might account for the comorbidity between disordered gambling behavior and other substance-related addictive disorders. Longitudinal population-based study. A complete birth cohort of young adults born in Dunedin, New Zealand, between April 1, 1972, and March 31, 1973 (N = 939; 475 men, 464 women). Multidimensional Personality Questionnaire assessments of personality were obtained at age 18 years; structured interview-based diagnoses of past-year problem gambling and alcohol, cannabis, and nicotine dependence were obtained at age 21 years. Problem gambling at age 21 years was associated with higher scores on the higher-order personality dimension of negative emotionality (d = 0.90) and with lower scores on the personality dimension of constraint (d = -0.72) measured at age 18 years compared with control subjects who did not have a past-year addictive disorder at age 21 years. Problem gambling was also associated with Multidimensional Personality Questionnaire indicators of risk-taking (d = 0.50) and impulsivity (d = 0.56). The personality profile associated with problem gambling was similar to the profiles associated with alcohol, cannabis, and nicotine dependence. The relations between problem gambling and the substance-related addictive disorders (odds ratios = 3.32-3.61) were reduced after controlling for individual differences in personality (odds ratios = 1.90-2.32). From the perspective of personality, problem gambling has much in common with the addictive disorders, as well as with the larger class of "externalizing" or "disinhibitory" disorders. Knowledge gained from the study of common personality underpinnings may be helpful in determining where disordered gambling behavior should reside in our diagnostic classification system.


Purification of messenger ribonucleoprotein particles from rabbit reticulocytes by zonal centrifugation in metrizamide.
A large-scale purification procedure for messenger ribonucleoprotein (mRNP) particles from rabbit reticulocyte polysomes is described. The mRNP particles were dissociated from polysomes by treatment with urea and separated by differential centrifugation under conditions of high ionic strength. Zonal centrifugation in a metrizamide buoyant density gradient was the final purification step. One major class of mRNA particle was observed. The RNA was defined as mRNA by polyacrylamide gel electrophoresis and by globin production in a cell-free protein-synthesizing system. The twenty-three different proteins associated with the particle were a discrete set of proteins, which ranged in molecular weight from 175,000 to 23,500. The relative amount of each peptide in the particle was determined from a gel scan of the stained protein by computer simulation. None of the polypeptides comigrated with proteins from the 40-S and 60-S ribosomal subunits when analyzed by two-dimensional polyacrylamide gel electrophoresis.


Seroxat and the suppression of clinical trial data: regulatory failure and the uses of legal ambiguity.
This article critically evaluates the Medicines and Healthcare products Regulatory Agency's announcement, in March 2008, that GlaxoSmithKline would not face prosecution for deliberately withholding trial data, which revealed not only that Seroxat was ineffective at treating childhood depression but also that it increased the risk of suicidal behaviour in this patient group. The decision not to prosecute followed a four and a half year investigation and was taken on the grounds that the law at the relevant time was insufficiently clear. This article assesses the existence of significant gaps in the duty of candour which had been assumed to exist between drugs companies and the regulator, and reflects upon what this episode tells us about the robustness, or otherwise, of the UK's regulation of medicines.


Magnetic resonance imaging of the normal canine thyroid gland.
The magnetic resonance (MR) imaging features of the normal canine thyroid gland were retrospectively compiled from images acquired in 44 dogs presented for a variety of diseases unrelated to the thyroid gland. The appearance of the thyroid gland on different sequences, including pre- and postcontrast T1-weighted, T2-weighted, two-dimensional gradient echo, three-dimensional T2*-weighted gradient echo and proton density weighted images, were described in different image planes. The characteristic shape, location, and intensity of thyroid lobes compared with surrounding structures made them easily detectable in all dogs. The most common location of the thyroid lobes was dorsolateral to the trachea with the maximal cross-sectional area of the lobes located ventral to C2/3 or C3 in more than 85% of the dogs. The majority of the lobes were ovoid on transverse images. An isthmus was seen in one large dog and parathyroid glands could not be seen. The mean maximal thyroid lobe diameter on transverse images was 8.1 mm, being twice the mean diameter of the common carotid artery. Considering the excellent conspicuity and characteristic appearance of the canine thyroid gland, MR imaging can be beneficial in the diagnosis of diffuse thyroid diseases, in differentiating thyroid vs. nonthyroid neck masses and in staging and treatment planning of thyroid tumors in this species.


QUADAS-2: a revised tool for the quality assessment of diagnostic accuracy studies.
In 2003, the QUADAS tool for systematic reviews of diagnostic accuracy studies was developed. Experience, anecdotal reports, and feedback suggested areas for improvement; therefore, QUADAS-2 was developed. This tool comprises 4 domains: patient selection, index test, reference standard, and flow and timing. Each domain is assessed in terms of risk of bias, and the first 3 domains are also assessed in terms of concerns regarding applicability. Signalling questions are included to help judge risk of bias. The QUADAS-2 tool is applied in 4 phases: summarize the review question, tailor the tool and produce review-specific guidance, construct a flow diagram for the primary study, and judge bias and applicability. This tool will allow for more transparent rating of bias and applicability of primary diagnostic accuracy studies.


Estimation of effective plant area index for South Korean forests using LiDAR system.
Light Detection and Ranging (LiDAR) systems can be used to estimate both vertical and horizontal forest structure. Woody components, the leaves of trees and the understory can be described with high precision, using geo-registered 3D-points. Based on this concept, the Effective Plant Area Indices (PAI(e)) for areas of Korean Pine (Pinus koraiensis), Japanese Larch (Larix leptolepis) and Oak (Quercus spp.) were estimated by calculating the ratio of intercepted and incident LIDAR laser rays for the canopies of the three forest types. Initially, the canopy gap fraction (G ( LiDAR )) was generated by extracting the LiDAR data reflected from the canopy surface, or inner canopy area, using k-means statistics. The LiDAR-derived PAI(e) was then estimated by using G ( LIDAR ) with the Beer-Lambert law. A comparison of the LiDAR-derived and field-derived PAI(e) revealed the coefficients of determination for Korean Pine, Japanese Larch and Oak to be 0.82, 0.64 and 0.59, respectively. These differences between field-based and LIDAR-based PAI(e) for the different forest types were attributed to the amount of leaves and branches in the forest stands. The absence of leaves, in the case of both Larch and Oak, meant that the LiDAR pulses were only reflected from branches. The probability that the LiDAR pulses are reflected from bare branches is low as compared to the reflection from branches with a high leaf density. This is because the size of the branch is smaller than the resolution across and along the 1 meter LIDAR laser track. Therefore, a better predictive accuracy would be expected for the model if the study would be repeated in late spring when the shoots and leaves of the deciduous trees begin to appear.


Evaluation of a multiplex polymerase chain reaction assay for simultaneous detection of Rhodococcus equi and the vapA gene.
To evaluate sensitivity and specificity of a multiplex polymerase chain reaction (PCR) assay for simultaneous detection of Rhodococcus equi and differentiation of strains that contain the virulence-associated gene (vapA) from strains that do not. 187 isolates of R equi from equine and nonequine tissue and environmental specimens and 27 isolates of bacterial species genetically or morphologically similar to R equi. The multiplex PCR assay included 3 gene targets: a universal 311-bp bacterial 16S ribosomal RNA amplicon (positive internal control), a 959-bp R equi-specific target in the cholesterol oxidase gene (choE), and a 564-bp amplicon of the vapA gene. Duplicate multiplex PCR assays for these targets and confirmatory singleplex PCR assays for vapA and choE were performed for each R equi isolate. An additional PCR assay was used to examine isolates for the vapB gene. Results of duplicate multiplex and singleplex PCR assays were correlated in all instances, revealing high specificity and reliability (reproducibility) of the vapA multiplex assay. Of the pulmonary isolates from horses with suspected R equi pneumonia, 97.4% (76/78) yielded positive results for vapA. Seven of 50 (14%) human isolates of R equi yielded positive results for vapA. Six human R equi isolates and 1 porcine isolate yielded positive results for vapB. No isolates with vapA and vapB genes were detected. The multiplex PCR assay is a sensitive and specific method for simultaneous confirmation of species identity and detection of the vapA gene. The assay appeared to be a useful tool for microbiologic and epidemiologic diagnosis and research.


Tyrosine phosphorylation of an interleukin 2 receptor-like protein in cells transformed by human T cell leukemia virus type I.
Three interleukin 2 (IL-2)-independent human T cell lines transformed with human T cell leukemia virus type I were analyzed for o-phosphotyrosine-containing proteins (Ptyr proteins). Membrane and intracellular immunofluorescence was positive with antibody to Ptyr (Ptyr antibody). 10 size classes of Ptyr proteins with molecular masses of 81-28 kD were isolated with Ptyr antibodies. Among these, proteins of 64 kD (pI 4.5 and 4.8-5.3) and 45 kD (pI 4.3) were found on the outer cell surface. The Ptyr protein of 64 kD (pI 4.5) had the characteristics of the IL-2 receptor (IL-2-R) in that it was recognized by two monoclonal antibodies directed against different epitopes on the IL-2-R.


Tracking eye gaze during interpretation of endoluminal three-dimensional CT colonography: visual perception of experienced and inexperienced readers.
To identify and compare key stages of the visual process in experienced and inexperienced readers and to examine how these processes are used to search a moving three-dimensional ( 3D three-dimensional ) image and their relationship to false-negative errors. Institutional review board research ethics approval was granted to use anonymized computed tomographic (CT) colonographic data from previous studies and to obtain eye-tracking data from volunteers. Sixty-five radiologists (27 experienced, 38 inexperienced) interpreted 23 endoluminal 3D three-dimensional CT colonographic videos. Eye movements were recorded by using eye tracking with a desk-mounted tracker. Readers indicated when they saw a polyp by clicking a computer mouse. Polyp location and boundary on each video frame were quantified and gaze data were related to the polyp boundary for each individual reader and case. Predefined metrics were quantified and used to describe and compare visual search patterns between experienced and inexperienced readers by using multilevel modeling. Time to first pursuit was significantly shorter in experienced readers (hazard ratio, 1.22 [95% confidence interval: 1.04, 1.44]; P = .017) but other metrics were not significantly different. Regardless of expertise, metrics such as assessment, identification period, and pursuit times were extended in videos where polyps were visible on screen for longer periods of time. In 97% (760 of 787) of observations, readers correctly pursued polyps. Experienced readers had shorter time to first eye pursuit, but many other characteristics of eye tracking were similar between experienced and inexperienced readers. Readers pursued polyps in 97% of observations, which indicated that errors during interpretation of 3D three-dimensional CT colonography in this study occurred in either the discovery or the recognition phase, but rarely in the scanning phase of radiologic image inspection.


Alloderm Covering Over Titanium Cranioplasty May Minimize Contour Deformities in the Frontal Bone Position.
Titanium cranioplasty is commonly used for surgical closure of skull defects post craniectomy. Superficial implantation of the mesh can result in discomfort, palpability, and in extreme patients, exposure of the mesh. Exposed titanium mesh can be complicated by infections and often requires implant revision or removal. Generally, the contour of the titanium mesh is camouflaged in the hairline of the patient and any aesthetic complication can remain inconspicuous. However, in the frontal bone position the thin hairless forehead skin often may not easily hide the contour of the underlying titanium mesh. The goal of this study was to demonstrate the usage of an alloderm covering over the titanium cranioplasty to possibly minimize the contour irregularities of titanium. Our index patient, a 22-year-old woman, was operated for left frontal craniectomy for frontal bone tumor extending to brain parenchyma. This resulted in a surgical defect that was repaired with titanium mesh cranioplasty and dural patch. It was felt that the patient would potentially feel and see the titanium mesh cranioplasty under the thin frontalis muscle. Therefore, over top of the titanium mesh a remnant piece of AlloDerm unused after the dural patch was placed to buffer the potential contour deformity. Postoperatively, the contour appears natural and the patient does not report any complaints of discomfort or mesh palpability. In a thin-skinned patient undergoing titanium mesh cranioplasty, the risk of mesh palpability or exposure can be significant, especially in areas of non-hair-bearing scalp and the protruding areas of the skull. Alloderm covering over the titanium mesh can provide improved aesthetic outcomes by minimizing contour deformity and may serve as an additional buffer in thin scalp.