Circulating non-hematological cells during cardiopulmonary bypass: new findings in cardiac surgery procedures.
Several factors have been historically advocated to explain the coagulative and inflammatory disorders following cardiopulmonary bypass (CPB). In this paper, we describe the presence of circulating non-hematological cells, introduced within the bloodstream during CPB. We defined the origin of the cells and tested their impact on coagulation. We collected peripheral arterial blood samples in twenty consecutive coronary artery bypass graft cases at four different surgical moments and assessed the presence and nature of circulating cells with the use of the CELLSEARCH® Test, immunocytochemistry and immunofluorescence, evaluating the expression of cytokeratin and calretinin. The effect of the circulating non-hematological cells on coagulation was tested in vitro, using the ROTEM assay. A mean of 263.85 ± 57.5 (median 258.5) cells were present in the samples following the suction of blood from the surgical field while all the other samples were negative (zero cells) (p<0.00001). Immunologic tests confirmed the mesothelial origin of the cells. The ROTEM® assay of the blood samples contaminated by the mesothelial cells presented longer clotting times (53.4 ± 8.2 secs 48.3 ± 8.9 sec, p=0.05), longer clot formation times (137.1 ± 31.5 sec vs 111.9 ± 25.2 sec, p=0.009), smaller alfa angle amplitudes (66.7 ± 9.1° vs 71.1 ± 5.1°, p=0.04) and maximum clot firmness times (59.0 ± 5.4 sec vs 61.9 ±4.6 sec, p=0.004) than the controls. The presence of circulating non-hematological cells during CPB with a mesothelial immunophenotype alters in vitro coagulation assays. This finding can help to further understand the pathophysiology of CPB.


Estimating the frequency of inpatient adverse events using a two-step retrospective chart review: a study protocol.
Adverse events are a major cause of patient harm in the hospitalised setting. Low-income and middle-income countries account for a disproportionate share of the global burden of adverse events. However, patient safety research is still centred around high-income countries and high-resource health systems. The methods and data produced from these efforts are ill-suited to low-income and middle-income systems due to the social and technical differences between these settings. We aim to use our pilot-tested, locally developed methodology to estimate the frequency and characteristics of adverse events in hospitalised patients in a lower-middle-income country to inform patient safety policies and initiatives. This multi-centre study will employ a two-step chart review methodology to identify adverse events in a representative sample of patients admitted at five hospitals between 1 January 2019 and 31 December 2019. The first step will include assessing patient files against a list of triggers to detect adverse events and the second step will involve an in-depth review of the events to capture pertinent characteristics. The triggers have been adapted from validated tools used in other studies. The reviewing team will be trained on the use of research tools and operational definitions to ensure that data are collected uniformly. The main outcome of interest is the rate at which adverse events occur in hospitalised patients. Further analysis will look to identify and quantify associations between the main outcome of interest and a variety of variables such as patient age and gender using tests of independence and regression techniques. This study protocol has been approved by the Ethics Review Committee at Aga Khan University (Reference number: 2023-6324-24566). The findings of this study will be published in a peer-reviewed journal and disseminated to the public through national and international conferences, workshops, websites and social media.


Bone Marrow Mesenchymal Stem Cells Attenuate Mitochondria Damage Induced by Hypoxia in Mouse Trophoblasts.
We aimed to observe the change of mitochondrial function and structure as well as the cell function induced by hypoxia in mouse trophoblasts, and moreover, to validate the restoration of these changes after co-culture with bone marrow mesenchymal stem cells (hereinafter referred to as "MSCs"). Further, we explored the mechanism of MSCs attenuating the functional damage of trophoblasts caused by hypoxia. Cells were divided into two groups, trophoblasts and MSCs+trophoblasts respectively, and the two groups of cells were incubated with normoxia or hypoxia. Chemiluminescence was used to assay the β-HCG and progesterone in cell culture supernatants quantitatively. Western blotting and PCR were applied to detect the expression of Mfn2, MMP-2, MMP-9 and integrin β1 in the two groups. The mitochondrial membrane potential of each group of cells was detected with JC-1 dye and the ATP content was measured by the phosphomolybdic acid colorimetric method. We utilized transmission electron microscopy for observing the ultrastructure of mitochondria in trophoblasts. Finally, we assessed the cell apoptosis with flow cytometry (FCM) and analyzed the expression of the apoptosis related genes-Bcl-2, Bax, Caspase3 and Caspase9 by western blotting. The results showed that the Mfn2 expression was reduced after 4 h in hypoxia compared with that in normoxia, but increased in the co-culture group when compared with that in the separated-culture group (p<0.05). In addition, compared with the separated-culture group, theβ-HCG and progesterone levels in the co-culture group were significantly enhanced (p<0.05), and so were the expressions of MMP-2, MMP-9 and integrin β1 (p<0.05). Moreover, it exhibited significantly higher in ATP levels and intensified about the mitochondrial membrane potential in the co-culture group. TEM revealed disorders of the mitochondrial cristae and presented short rod-like structure and spheroids in hypoxia, however, in the co-culture group, the mitochondrial cristae had a relatively regular arrangement and the mitochondrial ultrastructure showed hyperfusion. The expression of Bax, Caspase3 and Caspase9 was decreased in the co-culture group when compared with that in trophoblast cells cultured alone (p<0.05), while the Bcl-2 levels and the Apoptosis Index (AI) were markedly increased in the co-culture group (p<0.05). Bone marrow mesenchymal stem cells can attenuate mitochondria damage and cell apoptosis induced by hypoxia; the mechanism could be upregulating the expression of Mfn2 in mouse trophoblasts and changing mitochondrial structure.


[IBD ahead 2010--Answering important questions in Crohn's disease treatment].
The treatment of patients with inflammatory bowel disease has become more complex in recent years through the introduction of various immunosuppressive agents as well as the approval of monoclonal antibodies against TNF-α and patients receiving such treatment must be carefully monitored. National and international guidelines define a diagnostic and therapeutic context for the practitioner, but can only partially respond to specific questions on the procedure for individual patients. Within the framework of a project initiated by Abbott entitled "IBD ahead" 38 German IBD experts have elaborated concrete proposals for dealing with corticosteroids, immunosuppressants and TNF-α antibodies on the basis of the published literature and their own personal experience in order to close the gap between these guidelines and daily clinical practice. Statements were developed on the choice of correct timing of initiation, dose and duration of the individual substances and on how to proceed with patients exhibiting treatment failure. Moreover, recommendations are also made on drug combination strategies, safety monitoring and the risks regarding the development of infectious complications and malignancies. These recommendations are illustrated by case studies from everyday practice in participating centres.


Manganese lipoxygenase oxidizes bis-allylic hydroperoxides and octadecenoic acids by different mechanisms.
Manganese lipoxygenase (MnLOX) oxidizes (11R)-hydroperoxylinolenic acid (11R-HpOTrE) to a peroxyl radical. Our aim was to compare the enzymatic oxidation of 11R-HpOTrE and octadecenoic acids with LOO-H and allylic C-H bond dissociation enthalpies of ~88 and ~87kcal/mol. Mn(III)LOX oxidized (11Z)-, (12Z)-, and (13Z)-18:1 to hydroperoxides with R configuration, but this occurred at insignificant rates (<1%) compared to 11R-HpOTrE. We next examined whether transitional metals could mimic this oxidation. Ce(4+) and Mn(3+) transformed 11R-HpOTrE to hydroperoxides at C-9 and C-13 via oxidation to a peroxyl radical at C-11, whereas Fe(3+) was a poor catalyst. Our results suggest that MnLOX oxidizes bis-allylic hydroperoxides to peroxyl radicals in analogy with Ce(4+) and Mn(3+). The enzymatic oxidation likely occurs by proton-coupled electron transfer of the electron from the hydroperoxide anion to Mn(III) and H(+) to the catalytic base, Mn(III)OH(-). Hydroperoxides abolish the kinetic lag times of MnLOX and FeLOX by oxidation of their metal centers, but 11R-HpOTrE was isomerized by MnLOX to (13R)-hydroperoxy-(9Z,11E,15Z)-octadecatrienoic acid (13R-HpOTrE) with a kinetic lag time. This lag time could be explained by two competing transformations, dehydration of 11R-HpOTrE to 11-ketolinolenic acid and oxidation of 11R-HpOTrE to peroxyl radical; the reaction rate then increases as 13R-HpOTrE oxidizes MnLOX with subsequent formation of two epoxyalcohols. We conclude that oxidation of octadecenoic acids and bis-allylic hydroperoxides occurs by different mechanisms, which likely reflect the nature of the hydrogen bonds, steric factors, and the redox potential of the Mn(III) center.


Perceived floor slipperiness and floor roughness in a gait experiment.
Slips and falls contribute to occupational injuries and fatalities globally. Both floor slipperiness and floor roughness affect the occurrence of slipping and falling. Investigations on fall-related phenomena are important for the safety and health of workers. The purposes of this study were to: compare the perceived floor slipperiness before and after walking on the floor; compare the perceived floor slipperiness with and without shoes for males and females; discuss the perceived floor roughness based on barefoot walking; and establish regression models to describe the relationship between perceived floor slipperiness and actual friction of the floors. Male and female subjects walked on 3 m walkways with or without shoes. The perceived floor slipperiness ratings both before and after their walk were collected. The perceived floor slipperiness both before and after walking were significantly affected by both floor and surface conditions. Gender, floor, surface, and footwear conditions were all significant factors affecting the adjustment of perceived floor slipperiness. The subjects made more adjustment on perceived floor slipperiness rating when they had shoes on than when they were barefooted. Regression models were established to describe the relationship between perceived floor slipperiness and floor coefficient of friction. These models may be used to estimate perceived floor slipperiness, or in reverse, the coefficient of friction of the floor, so as to prevent slipping and falling in workplaces.


Method for assessing lead, cadmium, mercury and arsenic in high-density polyethylene packaging and study of the migration into yoghurt and simulant.
The purpose of this paper was to assess the concentration of lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As) in high-density polyethylene (HDPE) packaging intended for contact with yoghurt and the migration of these elements using the food itself and 3% acetic acid as a food simulant in accordance to ANVISA, the Brazilian Health Surveillance Agency. In order to perform this study, it was necessary to develop and validate a method by inductively coupled plasma optical emission spectroscopy (ICP-OES) analysis. For method validation, the parameters linearity, limits of detection (LODs) and quantification (LOQs), accuracy and precision were determined. Fifteen commercial samples of yoghurt, marketed in Campinas - São Paulo (Brazil), were used for the analysis. The packaging and yoghurt were digested in high-pressure ashing equipment (HPA) and the migration of the elements into simulant were determined directly in the solution. The validated method proved adequate and the results obtained showed that all the packaging had levels of Hg and Cd below the LOQ, corresponding to 1.0 and 1.5 μg l(-1), respectively. The highest levels of As and Pb were 0.87 and 462.3 mg kg(-1), respectively. The migration of these elements to the yoghurt after 45 days of contact at 4ºC was below the LOQ for all the samples assessed. The results of specific migration into 3% acetic acid simulant showed the concentrations of Cd, Hg and As below 5, 5 and 10 µg kg(-1), respectively, which are the maximum limits set by ANVISA. However, for three samples the packaging lid showed migration of Pb into simulant ranging from 30.6 to 40.2 μg kg(-1), exceeding the limit set by ANVISA of 10 μg kg(-1).


Mathematical model of Ehrlichia chaffeensis transmission dynamics in dogs.
Ehrlichia chaffeensis is a tick-borne disease transmitted by ticks to dogs. Few studies have mathematical modelled such tick-borne disease in dogs, and none have developed models that incorporate different ticks' developmental stages (discrete variable) as well as the duration of infection (continuous variable). In this study, we develop and analyze a model that considers these two structural variables using integrated semigroups theory. We address the well-posedness of the model and investigate the existence of steady states. The model exhibits a disease-free equilibrium and an endemic equilibrium. We calculate the reproduction number (T0). We establish a necessary and sufficient condition for the bifurcation of an endemic equilibrium. Specifically, we demonstrate that a bifurcation, either backward or forward, can occur at T0=1, leading to the existence, or not, of an endemic equilibrium even when T0<1. Finally, numerical simulations are employed to illustrate these theoretical findings.


A new microdeletion syndrome of 5q31.3 characterized by severe developmental delays, distinctive facial features, and delayed myelination.
Chromosomal deletion including 5q31 is rare and only a few patients have been reported to date. We report here the first two patients with a submicroscopic deletion of 5q31.3 identified by microarray-based comparative genomic hybridization. The common clinical features of both patients were marked hypotonia,feeding difficulty in infancy, severe developmental delay, and epileptic/nonepileptic encephalopathy associated with delayed myelination. Both patients also shared characteristic facial features,including narrow forehead, low-set and dysmorphic ears, bilateral ptosis, anteverted nares, long philtrum, tented upper vermilion,edematous cheeks, and high arched palate. The deleted region contains clustered PCDHs, including PCDHA [corrected]. and PCDHG, which are highly expressed in the brain where they function to guide neurons during brain development, neuronal differentiation, and synaptogenesis. The common deletion also contains neuregulin 2(NRG2), a major gene for neurodevelopment. We suggest that 5q31.3 deletion is responsible for severe brain developmental delay and distinctive facial features, and that the common findings in these two patients should be recognized as a new microdeletion syndrome. We need further investigations to determine which genes are really responsible for patients' characteristic features


Physiological analysis to quantify training load in badminton.
To estimate the training load of specific on court training regimens based on the magnitude of variation of heart rate-lactate response during specific training and to determine the magnitude of variation of biochemical parameters (urea, uric acid, and creatine phosphokinase (CPK)) 12 hours after the specific training programme so as to assess training stress. The study was conducted on six national male badminton players. Maximum oxygen consumption (VO2), ventilation (VE), heart rate, and respiratory quotient were measured by a protocol of graded treadmill exercise. Twelve training sessions and 35 singles matches were analysed. Heart rate and blood lactate were monitored during technical training routines and match play. Fasting blood samples collected on two occasions--that is, during off season and 12 hours after specific training--were analysed for serum urea, uric acid, and CPK. Analysis of the on court training regimens showed lactate values of 8-10.5 mmol/l in different phases. The percentage of maximum heart rate ranged from 82% to 100%. Urea, uric acid, and CPK activity showed significant changes from (mean (SD)) 4.93 (0.75) mmol/l to 5.49 (0.84) mmol/l, 0.23 (0.04) to 0.33 (0.06) mmol/l, and 312 (211.8) to 363 (216.4) IU/l respectively. Maximum lactate reported in the literature ranges from 3-6 mmol/l. Comparatively high lactate values and high percentage of maximum heart rate found in on court training show a considerable stress on muscular and cardiovascular system. The training load needs appropriate monitoring to avoid over-training. Workouts that are too intensive may interfere with coordination, a factor that is important in sports requiring highly technical skill such as badminton.


Individual and community correlates of young people's high-risk drinking in Victoria, Australia.
To examine key individual- and community-level correlates of regular very high-risk drinking (more than 20 drinks at least 12 times a year for males and more than 11 drinks at least 12 times a year for females) among young (16-24) drinkers in Victoria. CATI survey of 10,879 young Victorian drinkers. Multi-level modelling of correlates of proportion drinking at very high-risk levels at both individual (e.g. age, gender) and community levels (e.g. alcohol outlet density, remoteness). One-fifth of the sample reported regular (at least monthly) very high-risk drinking. Significant individual-level correlates of very high-risk drinking included age, gender, cultural background, recreational spending money, life stage, living situation, family conflict and age at first drink. Significant community-level correlates included remoteness (living in a rural area) and packaged liquor outlet density. The study highlights a range of important factors relating to regular drinking of dangerous amounts of alcohol by youth. Interventions aimed at preventing early initiation or managing recreational spending should be explored. Further, regulatory management of packaged liquor outlets and or remote rural communities in Victoria with high levels of dangerous drinking should be a priority.


Properties and mechanisms of olfactory learning and memory.
Memories are dynamic physical phenomena with psychometric forms as well as characteristic timescales. Most of our understanding of the cellular mechanisms underlying the neurophysiology of memory, however, derives from one-trial learning paradigms that, while powerful, do not fully embody the gradual, representational, and statistical aspects of cumulative learning. The early olfactory system-particularly olfactory bulb-comprises a reasonably well-understood and experimentally accessible neuronal network with intrinsic plasticity that underlies both one-trial (adult aversive, neonatal) and cumulative (adult appetitive) odor learning. These olfactory circuits employ many of the same molecular and structural mechanisms of memory as, for example, hippocampal circuits following inhibitory avoidance conditioning, but the temporal sequences of post-conditioning molecular events are likely to differ owing to the need to incorporate new information from ongoing learning events into the evolving memory trace. Moreover, the shapes of acquired odor representations, and their gradual transformation over the course of cumulative learning, also can be directly measured, adding an additional representational dimension to the traditional metrics of memory strength and persistence. In this review, we describe some established molecular and structural mechanisms of memory with a focus on the timecourses of post-conditioning molecular processes. We describe the properties of odor learning intrinsic to the olfactory bulb and review the utility of the olfactory system of adult rodents as a memory system in which to study the cellular mechanisms of cumulative learning.


Predicting Impacts of Contact Tracing on Epidemiological Inference from Phylogenetic Data.
Robust sampling methods are foundational to many inference problems in the phylodynamic field, yet the impact of using contact tracing, a type of non-uniform sampling used in public health applications, is not well understood. To investigate and quantify how this non-uniform sampling method influences recovered phylogenetic tree structure, we developed a new simulation tool called SEEPS (Sequence Evolution and Epidemiological Process Simulator) that allows for the simulation of contact tracing and the resulting transmission tree, pathogen phylogeny, and corresponding virus genetic sequences. Importantly, SEEPS takes within-host evolution into account when generating pathogen phylogenies and sequences from transmission histories. Using SEEPS, we demonstrate that contact tracing can significantly impact the structure of the resulting tree as described by popular tree statistics. Contact tracing generates phylogenies that are less balanced than the underlying transmission process, less representative of the larger epidemiological process, and affects the internal/external branch length ratios that characterize specific epidemiological scenarios. We also examine a 2007-2008 Swedish HIV-1 outbreak and the broader 1998-2010 European HIV-1 epidemic to highlight the differences in contact tracing and expected phylogenies. Aided by SEEPS, we show that the Swedish outbreak was strongly influenced by contact tracing even after downsampling, while the broader European Union epidemic showed little evidence of universal contact tracing, agreeing with the known epidemiological information about sampling and spread. SEEPS is available at github.com/MolEvolEpid/SEEPS.


[DNA from cells infected with herpes simplex virus and from human tumors as an inducer of synthesis of a new nonvirion antigen].
DNA from herpes simplex virus type II (HSV-II)-infected cells isolated by using SDS, phenol, and pronase as well as by a new method of Pignatti et al. from triton X-100-NaCl-supernatant was shown to be capable to induce production of an early nonvirion (new) antigen in sensitive systems of Vero, RS-537 cells and others. It was also established that as a result of transfection of DNA recovered from 13 speciment of cancer tumors of the cervix and ovaries (9 tumors contained the new antigen and 4 were free from it), the new antigen associated with HSV-II was synthesized in 8 cases in the sensitive cells as early as 6 hours postinoculation whereas no viral antigens could be detected by this time. No viral antigens could be detected within 24 hours either, with the exception of 3 cases where CPE also developed by 24 hours indicating the appearance of the virus. DNA recovered from normal tissues of the cervix was incapable of inducing the new antigen.


Specifications of the variant curation guidelines for ITGA2B/ITGB3: ClinGen Platelet Disorder Variant Curation Panel.
Accurate and consistent sequence variant interpretation is critical to the correct diagnosis and appropriate clinical management and counseling of patients with inherited genetic disorders. To minimize discrepancies in variant curation and classification among different clinical laboratories, the American College of Medical Genetics and Genomics (ACMG), along with the Association for Molecular Pathology (AMP), published standards and guidelines for the interpretation of sequence variants in 2015. Because the rules are not universally applicable to different genes or disorders, the Clinical Genome Resource (ClinGen) Platelet Disorder Expert Panel (PD-EP) has been tasked to make ACMG/AMP rule specifications for inherited platelet disorders. ITGA2B and ITGB3, the genes underlying autosomal recessive Glanzmann thrombasthenia (GT), were selected as the pilot genes for specification. Eight types of evidence covering clinical phenotype, functional data, and computational/population data were evaluated in the context of GT by the ClinGen PD-EP. The preliminary specifications were validated with 70 pilot ITGA2B/ITGB3 variants and further refined. In the final adapted criteria, gene- or disease-based specifications were made to 16 rules, including 7 with adjustable strength; no modification was made to 5 rules; and 7 rules were deemed not applicable to GT. Employing the GT-specific ACMG/AMP criteria to the pilot variants resulted in a reduction of variants classified with unknown significance from 29% to 20%. The overall concordance with the initial expert assertions was 71%. These adapted criteria will serve as guidelines for GT-related variant interpretation to increase specificity and consistency across laboratories and allow for better clinical integration of genetic knowledge into patient care.


The capture-mark-recapture technique can be used as a stopping rule when searching in systematic reviews.
Researchers have no empirically based search stopping rule when looking for potentially relevant articles for inclusion in systematic reviews. We tested a stopping strategy based on capture-mark-recapture (CMR; i.e., the Horizon Estimate) statistical modeling to estimate the total number of articles in the domain of clinical decision support tools for osteoporosis disease management using four large bibliographic databases (Medline, EMBASE, CINAHL, and EBM reviews). Retrospective evaluation of the Horizon Estimate using a systematic review of randomized controlled trials (RCTs) at two levels of article screening: title and abstract (1,246 potentially relevant articles) and full text (42 potentially relevant articles). The CMR model suggests that the total number of potential articles was 1,838 for the first level of screening, and 49 for the full-text level. The four databases provided 68% of known articles for the first level of screening and 81% for full-text screening. The CMR technique can be used in systematic reviews to estimate the closeness to capturing the total body of literature on a given topic. More studies are needed to objectively determine the usefulness of Horizon Estimates as a stopping rule strategy for systematic review searching.


Combined Extinction and Absorption UV-Visible Spectroscopy as a Method for Revealing Shape Imperfections of Metallic Nanoparticles.
Metallic nanoparticle solutions are routinely characterized by measuring their extinction spectrum (with UV-vis spectroscopy). Theoretical predictions such as Mie theory for spheres can then be used to infer important properties, such as particle size and concentration. Here we highlight the benefits of measuring not only the extinction (the sum of absorption and scattering) but also the absorption spectrum (which excludes scattering) for routine characterization of metallic nanoparticles. We use an integrating sphere-based method to measure the combined extinction-absorption spectra of silver nanospheres and nanocubes. Using a suite of electromagnetic modeling tools (Mie theory, T-matrix, surface integral equation methods), we show that the absorption spectrum, in contrast to extinction, is particularly sensitive to shape imperfections such as roughness, faceting, or edge rounding. We study in detail the canonical case of silver nanospheres, where small discrepancies between experimental and calculated extinction spectra are still common and often overlooked. We show that this mismatch between theory and experiment becomes much more important when considering the absorption spectrum and can no longer be dismissed as experimental imperfections. We focus in particular on the quadrupolar localized plasmon resonance of silver nanospheres, which is predicted to be very prominent in the absorption spectrum but is not observed in our experiments. We consider and discuss a number of possible explanations to account for this discrepancy, including changes in the dielectric function of Ag, size polydispersity, and shape imperfections such as elongation, faceting, and roughness. We are able to pinpoint faceting and roughness as the likely causes for the observed discrepancy. A similar analysis is carried out on silver nanocubes to demonstrate the generality of this conclusion. We conclude that the absorption spectrum is in general much more sensitive to the fine details of a nanoparticle geometry, compared to the extinction spectrum. The ratio of extinction to absorption also provides a sensitive indicator of size for many types of nanoparticles, much more reliably than any observed plasmon resonance shifts. Overall, this work demonstrates that combined absorption-extinction measurements provide a much richer characterization tool for metallic nanoparticles.


Surgical Treatments and Long-Term Outcomes for Pediatric Patients With Lumbar Spinal Tumors.
Retrospective case‒control study. This study aimed to report the effects of surgical intervention on spinal stability recovery and to assess the long-term outcomes of children and adolescents with lumbar tumors. From January 2016 to June 2021, 42 pediatric patients with lumbar tumors were selected and separated into different groups based on the surgical method used (total en bloc resection (TER) group, n = 21; piecemeal resection (PR) group, n = 21; titanium mesh (TM) group n = 23; artificial vertebrae (AV) group n = 19). The clinicopathological characteristics, treatments and related outcomes were described in detail and compared between groups, with P value ≤.05 indicating statistically significant differences. The average follow-up duration was 24.89 months, and the mean age was 14.89 ± 2.41 years. There were no significant differences in the mean operation time, average blood loss, complication rate, or length of hospital stay between the groups. The ODI, VAS and JOA scores at the final follow-up (FF) were elevated after surgery in all groups. The FF local angular drift (LOD) and lumbar angular drift (LUD) were greater in the TM group than in the AV group (P = .03, P = .001). After surgery, pediatric patients with lumbar tumors can obtain satisfactory spinal stability, effective relief of pain symptoms and substantial improvements in neurological function. There was no significant difference in the invasiveness, safety or timeliness between the 2 surgical methods, so TER is recommended due to its low postoperative recurrence rate and good local control. Spinal fusion in the AV group resulted in better spinal stability.


An ensemble framework for clustering protein-protein interaction networks.
Protein-Protein Interaction (PPI) networks are believed to be important sources of information related to biological processes and complex metabolic functions of the cell. The presence of biologically relevant functional modules in these networks has been theorized by many researchers. However, the application of traditional clustering algorithms for extracting these modules has not been successful, largely due to the presence of noisy false positive interactions as well as specific topological challenges in the network. In this article, we propose an ensemble clustering framework to address this problem. For base clustering, we introduce two topology-based distance metrics to counteract the effects of noise. We develop a PCA-based consensus clustering technique, designed to reduce the dimensionality of the consensus problem and yield informative clusters. We also develop a soft consensus clustering variant to assign multifaceted proteins to multiple functional groups. We conduct an empirical evaluation of different consensus techniques using topology-based, information theoretic and domain-specific validation metrics and show that our approaches can provide significant benefits over other state-of-the-art approaches. Our analysis of the consensus clusters obtained demonstrates that ensemble clustering can (a) produce improved biologically significant functional groupings; and (b) facilitate soft clustering by discovering multiple functional associations for proteins. Supplementary data are available at Bioinformatics online.


Humoral immune responses in patients with Lyme neuroborreliosis.
The aim of this study was to analyze and compare the humoral immune responses in serum and cerebrospinal fluid (CSF) for 34 adult patients with clinically evident Lyme neuroborreliosis, 27 patients with clinically suspected Lyme neuroborreliosis, and 32 patients with tick-borne encephalitis. Additionally, we wanted to compare the findings of two methods for the detection of intrathecally synthesized borrelial antibodies: the IDEIA Lyme neuroborreliosis test using flagellar antigen and an approach based on the Liaison indirect chemiluminescence immunoassay using the OspC and VlsE antigens. Borrelial IgM and IgG antibodies were detected by at least one of these methods in the sera of 22/34 (64.7%) and 28/34 (82.4%) patients with clinically evident Lyme neuroborreliosis, respectively, and in the cerebrospinal fluid of 22/34 (64.7%) and 20/34 (58.8%) of these patients, respectively. Intrathecal synthesis of borrelial IgM and/or IgG was found in 19/34 (55.9%) patients: IgM in 17/34 (50%) patients and IgG in 15/34 (44.1%) patients. The relatively low proportion of intrathecal synthesis of borrelial antibodies and the high ratio of IgM positivity could be explained by the short duration of neurological disease as evidenced by reported symptoms (median, 10 days). Assessment of the humoral immune response in the sera and CSF of patients with early Lyme neuroborreliosis confirmed previous findings on the relationship between the duration of illness and the proportion of patients with detectable responses.


Deficit in switching between functional brain networks underlies the impact of multitasking on working memory in older adults.
Multitasking negatively influences the retention of information over brief periods of time. This impact of interference on working memory is exacerbated with normal aging. We used functional MRI to investigate the neural basis by which an interruption is more disruptive to working memory performance in older individuals. Younger and older adults engaged in delayed recognition tasks both with and without interruption by a secondary task. Behavioral analysis revealed that working memory performance was more impaired by interruptions in older compared with younger adults. Functional connectivity analyses showed that when interrupted, older adults disengaged from a memory maintenance network and reallocated attentional resources toward the interrupting stimulus in a manner consistent with younger adults. However, unlike younger individuals, older adults failed to both disengage from the interruption and reestablish functional connections associated with the disrupted memory network. These results suggest that multitasking leads to more significant working memory disruption in older adults because of an interruption recovery failure, manifest as a deficient ability to dynamically switch between functional brain networks.


Current understanding on biosynthesis of microbial polysaccharides.
The surfaces of almost all microbes are decorated with remarkable variations of polysaccharides such as O-antigen, capsular polysaccharides (CPS), and exopolysaccharides (EPS) in bacteria, lipoarabinomannans (LAM) in mycobacteria and lipophosphoglycan (LPG) in Leishmania. These polysaccharides play important roles in many biological processes, and they can function as the virulence determinants in the pathogens. The basic structures of these polysaccharides are known, but they show species-specificity or stage-specificity. For example, there are 186 O-serotypes and 80 capsular serotypes in E. coli. Despite the variation, the range of strategies used for the biosynthesis and assembly of these microbial polysaccharides is limited. Depending on the assembly and translocation mechanisms, O-antigen biosynthesis is subdivided into three pathways, of which the Wzy-dependent pathway is widely used not only in O-antigen, but also in CPS and EPS.


Glyphosate reduces urediniospore development and Puccinia psidii disease severity on Eucalyptus grandis.
Recent studies have shown the effects of glyphosate drift on decreasing rust intensity on Eucalyptus grandis plants. However, the effects of the herbicide on Puccinia psidii initial development are unknown. In this study the systemic action of glyphosate on rust severity was evaluated on Eucalyptus plants maintained under greenhouse conditions. Urediniospore germination and apressorium formation on detached leaves and on water agar medium, previously treated with glyphosate, were also evaluated. Rust severity and the number of urediniospores per leaf area were significantly reduced with increasing glyphosate doses, even on branches not directly treated with the herbicide, indicating a systemic effect of glyphosate on pathogen development. Similarly, higher glyphosate doses also reduced germination and apressorium formation on detached Eucalyptus leaves, regardless of the direct application of the product on the leaf limb or on the petiole base. Puccinia psidii urediniospore germination in water agar medium also decreased with increasing herbicide doses. Reductions in germination and apressorium formation of P. psidii urediniospores with increasing glyphosate dose indicate that a lower severity and intensity of the disease may perhaps be due to blockage of the shikimic acid pathway in the fungal metabolic system.


Effect of gonadectomy on hepatic, lung and intestinal mixed function monooxygenase enzymes in the rat.
Aryl hydrocarbon hydroxylase (AHH) and 7-ethoxycoumarin O-deethylase (ECD) activities of microsomes from liver, lungs, and intestine of control and gonadectomized male and female rats were examined. Estradiol-treated ovariectomized female rats and testosterone-treated castrated male rats were also employed. Castration decreased hepatic AHH and ECD activities by 50% and 27%, respectively, whereas ovariectomy had no effect on these enzymes. Testosterone treatment of castrated male animals returned AHH and ECD activities to control values. Estradiol treatment of the ovariectomized female rats had no effect on the activities of these two enzymes. Gonadectomy of either sex did not alter AHH or ECD activities in microsomes from lungs and intestinal mucosa. No sex differences in lung AHH activities or lung and intestinal ECD activities were noted. AHH activities of liver and intestinal mucosa were greater by 86% and 42%, respectively, in male rats as compared to the female rats. Castration decreased the hepatic cytochrome P-450 content by 15%, while ovariectomy had no effect. The hepatic cytochrome P-450 content of control male rats was 27% higher than in the females. Organ specific alterations of mixed function monooxygenase enzymes occur with castration of male animals but not following ovariectomy of female rats.


Quality assurance for a program of comprehensive care for older persons.
Quality assurance (QA) for comprehensive programs like the Program of All-inclusive Care for the Elderly (PACE) requires a special strategy. The assessment phase should be capable of looking across the usual subdivisions of care to recognize the contributions of various disciplines, and to focus on the effects of that care on the patient. Measures should thus include both problem-specific and patient-focused elements. The tracer technique which follows the care of specific problems provides an opportunity to look at both the process and outcomes of care. An outcomes focus which looks at patient functioning as well as condition-specific parameters can include specific sentinel events whose presence suggests untoward developments. Quality assurance implies more than assessment. It represents a commitment to act responsibly on the information obtained to improve the care rendered. It includes a strategy for proactive involvement where caregivers are prompted to consider pertinent information in a timely fashion, and a retrospective remedial approach where the data are analyzed and presented in a format that can be readily understood and which suggests next steps to improve care.


Invited review: aging and the cardiovascular system.
Aging is associated with complex and diversified changes of cardiovascular structure and function. The heart becomes slightly hypertrophic and hyporesponsive to sympathetic (but not parasympathetic) stimuli, so that the exercise-induced increases in heart rate and myocardial contractility are blunted in older hearts. The aorta and major elastic arteries become elongated and stiffer, with increased pulse wave velocity, evidence of endothelial dysfunction, and biochemical patterns resembling early atherosclerosis. The arterial baroreflex is sizably altered in aging, but different components are differentially affected: there is a definite impairment of arterial baroreceptor control of the heart but much better preserved baroreceptor control of peripheral vascular resistance. Alterations at the afferent, central neural, efferent, and effector organ portions of the reflex arch have been claimed to account for age-related baroreflex changes, but no conclusive evidence is available on this mechanistic aspect. Reflexes arising from cardiopulmonary vagal afferents are also blunted in aged individuals. The cardiovascular and reflex changes brought about by aging may have significant implications for circulatory homeostasis in health and disease.


Assessing bleeding in acute coronary syndrome using the Bleeding Academic Research Consortium definition.
The Bleeding Academic Research Consortium (BARC) definition was proposed to overcome the heterogeneity among the many bleeding definitions. The aim of this study-level meta-analysis was to explore the incidence of BARC-assessed bleeding in acute coronary syndrome (ACS) studies and to ascertain the relation between these events and variables related to bleeding risk. We searched the literature for studies that reported bleeding events according to BARC criteria in ACS patients. An analysis on heterogeneity between studies in bleeding reports was performed with I test. A meta-regression was conducted to explore the relation between different types of BARC bleedings and patient and procedural features. Nine studies were included in the analysis. Overall, BARC 2 rates were higher than BARC 3 or 5 rates (6.3 versus 2.6%). An extremely high level of heterogeneity was detected both for BARC 2 (I 99.3%) and BARC 3 or 5 (I 97.5%) bleedings. Increasing age [β coefficient 0.4% (0.2-0.6%); P < 0.001] and renal impairment [β coefficient 1 6.5% (1-32.1%); P = 0.037] were associated with increased BARC 3 or 5 rates, whereas the use of glycoprotein IIb/IIIa inhibitors was the only factor related to an increased incidence of BARC 2 bleeding [β coefficient 2 2.3% (5.5-39%); P = 0.009]. The high level of heterogeneity in BARC bleeding reports only partially explained by bleeding risk profile suggests that a regulatory guidance to properly evaluate bleedings and to estimate the risk--benefit in clinical trials investigating different antithrombotic treatments in ACS patients is needed.


Iraq/Afghanistan-era Veterans with back pain: Characteristics and predictors of compensation and pension award.
Approximately 440,000 U.S. Veterans receive compensation for back and/or neck conditions. Veterans Benefits Administration (VBA) criteria state that back compensation determinations are based on impaired back function and not comorbidity or lifestyle, but whether compensation is based solely on consideration of the lumbosacral region is unknown. In this study, we conducted a cross-sectional analysis of medical chart data from 178 post-9/11 Veterans applying for service connection for back pain in fiscal year 2012 at the Department of Veterans Affairs Connecticut Healthcare System. Altogether, 62% were noted to have impairment of back functioning and 74% were awarded compensation. Rates of comorbidities (obesity, depression, smoking, and illicit drug use) were high. In multivariate models predicting compensation awarded, only having an impaired back was associated with service-connected compensation. Pain was associated with extent of service connection, but this relationship was fully mediated by functional back impairment. No other measure (including work status) significantly predicted compensation. In summary, service connection was largely based on functional impairment, as called for in VBA criteria. Although pain and comorbidities undoubtedly affect day-to-day functioning, these factors were not independently related to service connection. Veterans present with many remediable conditions, and the service-connection evaluation may be an opportunity to engage them in treatment.


Squamous cell carcinoma manifesting as cutaneous cystic lesions.
Squamous cell carcinoma (SCC), a malignancy of epidermal keratinocytes, is the second most common cause of skin cancer in the United States. Our case represents an unusual variant of this common tumor. We report a clinical presentation of a case of SCC occurring as cutaneous cystic lesions on the face of an 87-year-old white woman with a medical history of multiple SCCs.


Intestinal brush border peptidases: activities in normal and abnormal peroral intestinal biopsy specimens.
Discriminating substrates were used to develop assays that accurately measure the activity of each of four brush border peptidases in peroral jejunal biopsy specimens. Seventy-five biopsy specimens, 43 normal and 32 abnormal, were assayed for aminopeptidase A (EC 184.108.40.206), aminopeptidase N (EC 188.8.131.52), and disaccharidases, and 37 of these were assayed for membrane glycylleucine peptidase and Zn2+-stable aspartyllysine peptidase. Peptidase levels in normal biopsy specimens from males and females did not differ. Only aminopeptidase A levels changed significantly with age. The mean +/- SD units per gram of protein for each enzyme in peroral biopsy specimens of normal intestine were aminopeptidase A 1.7 +/- 0.7 for children and 3.1 +/- 1.3 for adults, membrane glycylleucine peptidase 16.3 +/- 10.6, aminopeptidase N 35.5 +/- 13.2, and Zn2+-stable aspartyllysine peptidase 42.6 +/- 28.5. In abnormal biopsy specimens, levels of three of the peptidases and the disaccharidases were significantly decreased, 37% to 51%. In contrast, aminopeptidase N activity was decreased only 7% in the abnormal biopsy specimens. These results provide a basis for further studies investigating the role of brush border peptidases in normal digestion and various pathologic states, including the search for individuals with brush border peptidase deficiencies.


Occupied and unoccupied estrogen receptor during estrous cycle and pregnancy.
Occupied and unoccupied forms of estrogen receptor (Re) were measured in hamster uterus during the estrous cycle and pseudopregnancy with a new assay method employing pyridoxal 5'-phosphate. In the cycle, occupied Re increased in cytosol and nuclear fractions in parallel with increasing serum estradiol during the follicular phase and then dropped sharply in both compartments during preovulatory progesterone secretion. Unoccupied Re occurred primarily in cytosol with low levels present in nuclei. It is pertinent that similar concentrations of occupied Re accumulate in cytosol and nuclear fractions, suggesting that occupied Re is distributed between these compartments according to an equilibrium model. Progesterone disrupts Re distribution through a selective action on the nucleus to downregulate occupied Re as was shown by progesterone treatment of the proestrous hamster. Re is chronically downregulated in the progesterone-dominated uterus, and the occupied form of Re recovers rapidly after progesterone withdrawal. Thus progesterone downregulates the Re system under physiological conditions by a selective action on the occupied form of Re in the target cell nucleus.


Isolation of cDNA clones coding for human tissue factor: primary structure of the protein and cDNA.
Tissue factor is a membrane-bound procoagulant protein that activates the extrinsic pathway of blood coagulation in the presence of factor VII and calcium. lambda Phage containing the tissue factor gene were isolated from a human placental cDNA library. The amino acid sequence deduced from the nucleotide sequence of the cDNAs indicates that tissue factor is synthesized as a higher molecular weight precursor with a leader sequence of 32 amino acids, while the mature protein is a single polypeptide chain composed of 263 residues. The derived primary structure of tissue factor has been confirmed by comparison to protein and peptide sequence data. The sequence of the mature protein suggests that there are three distinct domains: extracellular, residues 1-219; hydrophobic, residues 220-242; and cytoplasmic, residues 243-263. Three potential N-linked carbohydrate attachment sites occur in the extracellular domain. The amino acid sequence of tissue factor shows no significant homology with the vitamin K-dependent serine proteases, coagulation cofactors, or any other protein in the National Biomedical Research Foundation sequence data bank (Washington, DC).


Identification of phenyldecanoic acid as a constituent of triacylglycerols and wax ester produced by Rhodococcus opacus PD630.
Phenyldecane supported growth and lipid accumulation of Rhodococcus opacus PD630 during cultivation under nitrogen-limiting conditions. The results of this study suggested that the hydrocarbon phenyldecane was degraded by monoterminal oxidation, followed by beta-oxidation of the alkyl side-chain to phenylacetic acid, and by an additional degradative route for the oxidation of the latter to intermediates of the central metabolism. alpha-Oxidation of phenyldecanoic acid also occurred to some extent. Phenyldecanoic acid, the monoterminal oxidation product, was also utilized for the biosynthesis of a novel wax ester and novel triacylglycerols. The formation of the wax ester phenyldecylphenyldecanoate probably resulted from the condensation of phenyldecanoic acid and phenyldecanol, which were produced as metabolites during the catabolism of phenyldecane. Two types of triacylglycerol were detected in phenyldecane-grown cells of strain PD630. Triacylglycerols containing only odd- and even-numbered aliphatic fatty acids, as well as triacylglycerols in which one fatty acid was replaced by a phenyldecanoic acid residue, occurred. Other phenyl intermediates, such as phenylacetic acid, phenylpropionic acid, 4-hydroxyphenylpropionic acid, protocatechuate and homogentisic acid, were excreted into the medium during cultivation on phenyldecane. On the basis of the results obtained, pathways for the catabolism and assimilation of phenyldecane by R. opacus PD630 are discussed.


Readmissions After Acute Myocardial Infarction: How Often Do Patients Return to the Discharging Hospital?
Background When patients require readmission after a recent myocardial infarction (MI), returning to the discharging (index) hospital may be associated with better outcomes as a result of greater continuity in care. However, little evidence exists to answer this frequent patient question. Methods and Results Among Medicare patients aged ≥65 years discharged home alive post-MI from 491 US hospitals in the ACTION (Acute Coronary Treatment Intervention Outcomes Network) Registry, we compared reason for readmission, duration of rehospitalization, and 30-day mortality between patients readmitted to the index versus nonindex hospital within 30 days of index MI discharge. Among 53 471 MI patients, 7715 (14%) were readmitted within 30 days, and most readmitted patients (73%) returned to the discharging hospital. Reason for readmission was not significantly associated with location of readmission. In multivariable modeling, the strongest factors associated with readmission to a nonindex hospital were distance from the discharging hospital, transfer-in during the index MI hospitalization, and frequency of nonindex hospital admissions in the year preceding to the index MI. Duration of rehospitalization did not differ significantly between patients readmitted to the index versus nonindex hospital (median, 4 versus 3 days; P=0.17). Mortality risk was also not significantly different between patients readmitted to the index versus nonindex hospital overall (7.4 versus 7.7%; adjusted odds ratio, 0.89; 95% CI, 0.73-1.10) and when stratified by reason for readmission (P for interaction=0.61). Conclusions Post-MI readmissions did not differ in reason for readmission, duration of rehospitalization, or associated mortality when compared between patients who returned to the discharging hospital and those who sought care elsewhere.


Synthesis and biological activity of tris- and tetrakiscatecholate siderophores based on poly-aza alkanoic acids or alkylbenzoic acids and their conjugates with beta-lactam antibiotics.
New linear and tripodal tri-aza- and tetra-aza alkanoic acids or alkylbenzoic acids were prepared as basic structures for siderophore mimetics from polyamines and oxocarbonic acids or formylbenzoic acids by catalytic hydrogenation. From these acids acetylated tris- and tetrakiscatecholates or 8-acyloxy-2,4-dioxo-benzoxazine derivatives as well as compounds with spacer groups were synthesized. These derivatives were coupled with ampicillin, amoxicillin, bacampicillin or cefaclor to new siderophore antibiotic conjugates. Most of the catecholate derivatives showed high siderophore activities in strains of Pseudomonas aeruginosa and Escherichia coli in a growth promotion assay under iron limitation conditions. The beta-lactam conjugates were highly active in vitro against Gram-negative bacteria correlating to the siderophore activity of the catecholate moiety and depending on the beta-lactam part. One ampicillin conjugate based on 5-(aminoethyl)-2,5,8-triazaalkylbenzoic acid was highly active against Gram-negative and Gram-positive bacteria. It was shown that conjugates with enhanced activity against Gram-negative bacteria use active iron uptake routes to penetrate the bacterial outer membrane barrier. Correlations between structure and biological activity were studied.


Synthesis and antiviral evaluation of 1,4-dioxane nucleoside analogues related to nucleoside dialdehydes.
Since biologically active nucleoside 2',3'-dialdehydes exist as six-membered cyclic acetals (1) in solution, we have investigated the antiviral activity of some structurally similar 1,4-dioxane nucleoside analogues. By reacting 2',3'-seconucleoside tosylates with base, the guanine (10) and adenine (18) substituted (hydroxymethyl)dioxanes have been constructed. In addition, an unusual adenine-substituted divinyl ether (22) was synthesized via a base-catalyzed, double elimination of a 2',3'-di-O-tosyl-2',3'-secoadenosine. None of these compounds showed significant antiviral activity.


Anti-Austerity Activity of Thai Medicinal Plants: Chemical Constituents and Anti-Pancreatic Cancer Activities of Kaempferia parviflora.
Human pancreatic tumor cells have an intrinsic ability to tolerate nutrition starvation and survive in the hypovascular tumor microenvironment, the phenomenon termed as "austerity". Searching for an agent that inhibits such tolerance to nutrient starvation and kills the pancreatic cancer cells preferentially in nutrient-starvation is a unique anti-austerity strategy in anti-cancer drug discovery. In this strategy, plant extracts and compounds are tested against PANC-1 human pancreatic cancer cell line under the conditions of nutrient-deprived medium (NDM) and nutrient-rich medium (DMEM), to discover the compounds that show selective cytotoxicity in NDM. Screening of twenty-five Thai indigenous medicinal plant extracts for their anti-austerity activity against the PANC-1 human pancreatic cancer cell line in nutrient deprived medium (NDM) resulted in the identification of four active plants, Derris scandens, Boesenbergia pandurata, Citrus hystrix, and Kaempferia parviflora, with PC50 values 0.5-8.9 µg/mL. K. parviflora extract also inhibited PANC-1 cancer cell colony formation. Phytochemical investigation of K. parviflora extract led to the isolation of fourteen compounds, including two polyoxygenated cyclohexanes (1 and 2), eleven flavonoids (3-13), and β-sitosterol (14). Stereochemical assignment of compound 1 was confirmed through X-ray analysis. All isolated compounds were tested for their preferential cytotoxicity against PANC-1 cells. Among them, 5-hydroxy-7-methoxyflavone (3) displayed the most potent activity with a PC50 value of 0.8 µM. Mechanistically, it was found to induce apoptosis in PANC-1 cell death in NDM as evident by caspase cleavage. It was also found to inhibit PANC-1 cancer cell colony formation in DMEM. Therefore, compound 3 can be considered as a potential lead compound for the anticancer drug development based on the anti-austerity strategy.


Tailing DNA aptamers with a functional protein by two-step enzymatic reaction.
An efficient, quantitative synthetic strategy for aptamer-enzyme conjugates was developed by using a two-step enzymatic reaction. Terminal deoxynucleotidyl transferase (TdT) was used to first incorporate a Z-Gln-Gly (QG) modified nucleotide which can act as a glutamine donor for a subsequent enzymatic reaction, to the 3'-OH of a DNA aptamer. Microbial transglutaminase (MTG) then catalyzed the cross-linking between the Z-QG modified aptamers and an enzyme tagged with an MTG-reactive lysine containing peptide. The use of a Z-QG modified dideoxynucleotide (Z-QG-ddUTP) or a deoxyuridine triphosphate (Z-QG-dUTP) in the TdT reaction enables the controlled introduction of a single or multiple MTG reactive residues. This leads to the preparation of enzyme-aptamer and (enzyme)n-aptamer conjugates with different detection limits of thrombin, a model analyte, in a sandwich enzyme-linked aptamer assay (ELAA). Since the combination of two enzymatic reactions yields high site-specificity and requires only short peptide substrates, the methodology should be useful for the labeling of DNA/RNA aptamers with proteins.


Tuberculous peritonitis--report of 30 cases and review of the literature.
In this retrospective study, clinical presentation of peritoneal tuberculosis in an area with a high prevalence of tuberculosis is discussed. Thirty cases diagnosed in an eight-year period are presented. The disease was 2.3 times more common in females. Eighty per cent of the patients were less than 40 years old. Excluding two cases of long duration (one year and three years), mean duration of the symptoms before admission was 2.3 months. Fever, abdominal pain, swelling and weight loss were the main symptoms. Ascites was clinically detected in 80 per cent of cases. Intermediate strength PPD tuberculin skin test was positive in 61 per cent of cases. In 57.7 per cent there was radiologic evidence of pleuropulmonary disease suggestive of tuberculosis. Fourteen of 15 ascitic fluids examined were exudative in nature with predominance of lymphocytes. In the majority of patients diagnosis was proven with laparotomy and laparoscopy (12 cases in each group). In the remainder, tissue for diagnosis was obtained through blind peritoneal needle biopsy, percutaneous biopsy of liver and peripheral lymph node biopsy (two cases with each method).


Patients' organizations in rare diseases and involvement in drug information: Illustrations with LMC France, the French Association of Chronic Myeloid leukemia.
There are few areas in medicine in which patient-parent advocacy groups play such a central and prominent role as in the field of rare diseases. As illustrated by the French Association of Chronic Myeloid Leukemia (LMC France), its contribution is not only focused on its role as participants or key informant related to clinical studies but also on drug information and drug safety. The discovery of tyrosine kinase inhibitors against the BCR-ABL1 oncogenic fusion protein has revolutionized the management of CML, becoming a chronic illness rather than a life-threatening disease. Because ensuring ongoing well-being requires some knowledge, LMC France has built, in synergy with healthcare professionals from its scientific council, a CML-specific, patient-friendly knowledge base including resources and knowledge related to drug information (drug development, generics, pharmacovigilance) and drug safety using several educational tools such as videos, CML support, and CML drug sheets. To disseminate more largely, an e-university learning for regional key informants from LMC France and also CML patients and their caregivers was launched, including a large resources related to drug information and drug safety in synergy with hematologists and clinical pharmacologists.


Pharmacokinetics and cellular uptake of imatinib and its main metabolite CGP74588.
Despite the remarkable clinical response rates to imatinib in the treatment of bcr-abl leukemic patients, pharmacokinetic data on this relatively novel substance are needed to improve our understanding of the emergence of resistance, the interindividual variations of clinical response and the clinical and biologic relevance of its main metabolite N-desmethyl-imatinib. We present here pharmacokinetic data obtained with a newly designed HPLC approach in 97 patients with chronic myeloid leukemia or acute lymphatic leukemia (ALL) under treatment with imatinib that allowed us to calculate the AUC (39.5 microg.h/ml for an oral dose of 400 mg daily), the t(1/2) (18.2 h) and the peak concentration (1.92 micro/ml for an oral dose of 400 mg daily) of imatinib in plasma. In a subgroup of patients, the same parameters were analyzed for N-desmethyl-imatinib. We also provide data on the imatinib concentration in the cerebrospinal fluid (CSF) of ALL patients and demonstrate that oral administration of imatinib resulted only in a marginal flux across the blood-brain barrier. Finally, in an in vitro setting, we determined cellular concentrations of imatinib in HL-60 cells and showed an over-proportional uptake both in RPMI medium and in human plasma. Using an arithmetical approach combining all parameters obtained in imatinib-treated patients, we finally provide a conclusive approximation of basic pharmacokinetic data for both imatinib and its main metabolite N-desmethyl-imatinib.


Effects of starter nitrogen fertilizer on soybean root activity, leaf photosynthesis and grain yield.
The objective of this study was to examine the impact of starter nitrogen fertilizer on soybean root activity, leaf photosynthesis, grain yield and their relationship. To achieve this objective, field experiments were conducted in 2013 and 2014, using a randomized complete block design, with three replications. Nitrogen was applied at planting at rates of 0, 25, 50, and 75 kg N ha-1. In both years, starter nitrogen fertilizer benefited root activity, leaf photosynthesis, and consequently its yield. Statistically significant correlation was found among root activity, leaf photosynthetic rate, and grain yield at the developmental stage. The application of N25, N50, and N75 increased grain yield by 1.28%, 2.47%, and 1.58% in 2013 and by 0.62%, 2.77%, and 2.06% in 2014 compared to the N0 treatment. Maximum grain yield of 3238.91 kg ha-1 in 2013 and 3086.87 kg ha-1 in 2014 were recorded for N50 treatment. Grain yield was greater for 2013 than 2014, possibly due to more favorable environmental conditions. This research indicated that applying nitrogen as starter is necessary to increase soybean yield in Sangjiang River Plain in China.


Induction of indoleamine 2,3-dioxygenase (IDO) enzymatic activity contributes to interferon-gamma induced apoptosis and death receptor 5 expression in human non-small cell lung cancer cells.
Interferon-gamma (IFN-γ) has been used to treat various malignant tumors. However, the molecular mechanisms underlying the direct anti-proliferative activity of IFN-γ are poorly understood. In the present study, we examined the in vitro antitumor activity of IFN-γ on two human non-small-cell lung carcinoma (NSCLC) cell lines, H322M and H226. Our findings indicated that IFN-γ treatment caused a time-dependent reduction in cell viability and induced apoptosis through a FADD-mediated caspase-8/tBid/mitochondria-dependent pathway in both cell lines. Notably, we also postulated that IFN-γ increased indoleamine 2,3-dioxygenase (IDO) expression and enzymatic activity in H322M and H226 cells. In addition, inhibition of IDO activity by the IDO inhibitor 1-MT or tryptophan significantly reduced IFN-γ-induced apoptosis and death receptor 5 (DR5) expression, which suggests that IDO enzymatic activity plays an important role in the anti-NSCLC cancer effect of IFN-γ. These results provide new mechanistic insights into interferon-γ antitumor activity and further support IFN-γ as a potential therapeutic adjuvant for the treatment of NCSLC.


Electric field stimulation unmasks a subtle role for T-type calcium channels in regulating lymphatic contraction.
We previously identified two isoforms of T-type, voltage-gated calcium (Cav3) channels (Cav3.1, Cav3.2) that are functionally expressed in murine lymphatic muscle cells; however, contractile tests of lymphatic vessels from single and double Cav3 knock-out (DKO) mice, exhibited nearly identical parameters of spontaneous twitch contractions as wild-type (WT) vessels, suggesting that Cav3 channels play no significant role. Here, we considered the possibility that the contribution of Cav3 channels might be too subtle to detect in standard contraction analyses. We compared the sensitivity of lymphatic vessels from WT and Cav3 DKO mice to the L-type calcium channel (Cav1.2) inhibitor nifedipine and found that the latter vessels were significantly more sensitive to inhibition, suggesting that the contribution of Cav3 channels might normally be masked by Cav1.2 channel activity. We hypothesized that shifting the resting membrane potential (Vm) of lymphatic muscle to a more negative voltage might enhance the contribution of Cav3 channels. Because even slight hyperpolarization is known to completely silence spontaneous contractions, we devised a method to evoke nerve-independent, twitch contractions from mouse lymphatic vessels using single, short pulses of electric field stimulation (EFS). TTX was present throughout to block the potential contributions of voltage-gated Na+ channels in perivascular nerves and lymphatic muscle. In WT vessels, EFS evoked single contractions that were comparable in amplitude and degree of entrainment to those occurring spontaneously. When Cav1.2 channels were blocked or deleted, only small residual EFS-evoked contractions (~ 5% of normal amplitude) were present. These residual, EFS-evoked contractions were enhanced (to 10-15%) by the KATP channel activator pinacidil (PIN) but were absent in Cav3 DKO vessels. Our results point to a subtle contribution of Cav3 channels to lymphatic contractions that can be unmasked in the absence of Cav1.2 channel activity and when the resting Vm is more hyperpolarized than normal.


Group selection on a quantitative character.
We derive a model of group and individual selection on a quantitative character that is similar to the single-locus "metapopulation" models of group selection. Two alternative methods for the colonization of new or vacant habitats are examined and their effects are contrasted. In one model, all populations contribute migrants to a common pool, the "migrant pool," from which colonists are drawn at random to fill vacant sites. In the migrant pool there is complete mixing of individuals from different populations. This model of colonization is the one used in all previous models of group selection. In the other model, the "propagule pool" model, each propagule is made up of individuals derived from a single population and there is no mixing of colonists from different populations during propagule formation. The analysis shows that much more between-population genetic variance can be maintained with the propagule pool model than with the migrant pool model. Consequently, group selection can be much more effective in natural populations than is commonly supposed.


Illness Narratives of African Americans Living With Coronary Heart Disease: A Critical Interactionist Analysis.
How African American men and women respond to and manage living with coronary heart disease (CHD) is not well understood despite the well-documented disproportionate burden of CHD and its complications among African Americans in the United States. Through a critical interactionist perspective, we explore illness experiences of African Americans living with CHD and describe a broad range of micro-, meso-, and macro-contextual factors that influence their illness experiences. For participants in this study, CHD has become a "Black disease" wherein certain bodies have become historically and racially marked; a conceptualization maintained and passed on by African Americans themselves. Such findings highlight that CHD is more than a "lifestyle disease" where high-risk behaviors and lack of healthy choices are ultimate culprits. Rather, CHD is perceived by African Americans who have it as yet another product of ongoing racial and socio-structural dynamics through which their health burdens are created, sustained, and reproduced.


Social participation of employed and unemployed Israelis with disabilities.
The purpose of this study was to examine the relationship between employment and social participation among a random, national sample of 597 Israelis with disabilities of working age. Overall findings indicated that employed people with disabilities were significantly more integrated in social and civic activities than the unemployed. Whereas most of the unemployed attributed their restricted patterns of participation to the lack of adequate financial resources, accessible transportation, and encouragement from community organizations, the employed reported lack of time as their main barrier. These findings suggest that employment and rich community participation patterns might reinforce each other and thus serve as a potential vehicle for promoting social integration for people with disabilities in their communities.


Hepatitis B virus compartmentalization in the cerebrospinal fluid of HIV-infected patients.
We detected hepatitis B virus (HBV) DNA in the cerebrospinal fluid (CSF) of 26 adolescents co-infected with human immunodeficiency virus (HIV) and hepatitis B virus (HBV) with neurological disease and studied compartmentalization of HBV in the CSF. More than half of the subjects with positive HBV DNA plasma also had CSF positive for HBV. CSF HBV DNA was found in subjects with preserved blood-brain barrier integrity. In a subgroup of these subjects, compartmentalized evolution of HBV was demonstrated by distinct profiles of resistance mutations. Future studies are warranted to determine the clinical significance of HBV presence in the CSF and its contribution to HIV-associated neurological disease.


Antibody response to booster vaccination with tetanus and diphtheria in adults exposed to perfluorinated alkylates.
Recent studies suggest that exposure to perfluorinated alkylate substances (PFASs) may induce immunosuppression in humans and animal models. In this exploratory study, 12 healthy adult volunteers were recruited. With each subject, serum-PFAS concentrations were measured and their antibody responses prospectively followed for 30 days after a booster vaccination with diphtheria and tetanus. The results indicated that serum-PFAS concentrations were positively correlated and positively associated with age and male sex. The specific antibody concentrations in serum were increased from Day 4 to Day 10 post-booster, after which a constant concentration was reached. Serum PFAS concentrations showed significant negative associations with the rate of increase in the antibody responses. Interestingly, this effect was particularly strong for the longer-chain PFASs. All significant associations remained significant after adjustment for sex and age. Although the study involved a small number of subjects, these findings of a PFAS-associated reduction of the early humoral immune response to booster vaccination in healthy adults supported previous findings of PFAS immunosuppression in larger cohorts. Furthermore, the results suggested that cellular mechanisms right after antigen exposure should be investigated more closely to identify possible mechanisms of immunosuppression from PFAS.


Adult psychiatric and psychosocial outcomes of children with mild intellectual disability: a register follow-up of a population-based cohort.
Very few longitudinal psychiatric and psychosocial outcome studies of children with mild intellectual disability (MID) have been performed. The study group was population based and consisted of 82 individuals, born in 1979-1985 and diagnosed in childhood at ages between 3 and 15 years with MID. In the present study, register data regarding school attendance, employment, economic situation, psychiatric diagnoses and criminal sentences were retrieved for the years 1997-2018, when the individuals were up to 39 years old. At follow-up, data were obtained for 78 of the 82 individuals (47 male and 31 female). Mean age at follow-up was 36 years. Of the 78 individuals, 57 (73%) had exclusively received education for pupils with MID, but 21 (27%) had graduated from regular education of some sort (at least 9 years). Forty-four (56%) had never been employed, and 34 (44%) had been registered as employed for at least a shorter period. Forty-seven (60%) had received a sick pension at some point in adulthood. Of the 78 individuals, 44 (56%) had any psychiatric disorder recorded and about half of these (n = 21) had had inpatient treatment. A total of 31 of the 44 individuals in psychiatric care (70%) had ID noted as one of their diagnoses. Of the 78 individuals, 48 (62%) had support from the Act concerning Support and Service for Persons with Certain Functional Impairments (Swedish LSS law) as adults. Twenty-one individuals (27%) had had a criminal conviction, of whom five male individuals had been incarcerated. Individuals with MID constitute a heterogeneous group with regard to severity of functional impairment, co-occurring psychiatric disorders and need of support from society. Primary health care, psychiatry and habilitation services need to work together in order to meet the multiple needs of this group.


Hepatic zonal degeneration and necrosis in Reye syndrome.
Seven pediatric cases with hepatic peripheral zonal degeneration or necrosis in the liver, or both, were studied. From the standpoint of clinicopathological features, these cases fit best into the spectrum of Reye syndrome. Exogenous toxins, such as phosphorus and drugs, could not be implicated as principal causative factors in any of the cases. In the context of Reye syndrome, fatty acids would seem to have been the toxins most likely responsible for the pathogenesis of the peripheral zonal degeneration and necrosis in the liver.


Differential effects of 2-deoxy-D-glucose on surface glycoproteins of normal and transformed cells.
Sarcoma virus transformed rat kidney cells, NRK[MSV-MLV] cells, and their non-transformed counterparts, NRK cells, were repeatedly subcultured in medium supplemented with 1 mM 2-deoxy-D-glucose (dGlc). These dGlc-grown cells exhibited alterations in glycosylation of surface proteins. Gel filtration analyses of pronase-treated, trypsin-sensitive, surface fucopeptides indicated a reduction in the average size of the fucopeptides from both dGlc-grown NRK and NRK[MSV-MLV] cells. When the cells were grown in control medium the fucopeptides from the NRK[MSV-MLV] cells were larger than those from NRK cells, i.e. the fucopeptide difference observed in a wide range of transformed versus normal cells. The changes in the fucopeptides of dGlc-grown cells were greater for the NRK cells so that after growth in dGlc-supplemented medium there was an enhancement in the typical fucopeptide differences between normal and transformed cells, namely a larger difference in both size distribution and degree of sialylation. These dGlc-related alterations were readily reversed when the cells were transferred to control medium. The changes in glycosylation were compatible with continued cell replication although there was an increase in doubling time of the dGlc-grown cells, 6-7 h increase for NRK cells, 1-2 h increase for NRK[MSV-MLV] cells.


[Appendiceal carcinoid: report of 3 clinical cases and review of the literature].
The Authors report three cases of appendiceal carcinoids, found during appendectomy for acute appendicitis during the last 15 years. Appendiceal carcinoid is a rare neoplasm, more frequent in females age ranging between 20 and 29 years. Lack of symptoms explains the occasional diagnosis of this disease. The prognosis is related mostly to the size of the tumor, local and lymphatic invasion, presence of metastases, localization inside the appendix, histological type and age of the patient. Therapy is mainly surgical and consists of appendectomy and right colectomy according to different criteria which are being discussed in this article.


Respiratory and Ocular Symptoms Among Employees of an Indoor Waterpark Resort - Ohio, 2016.
In July 2015, a municipal health department in Ohio received complaints of respiratory and ocular symptoms from patrons of an indoor waterpark resort. In response, the health department conducted an online survey in August 2015 through which 19 (68%) patron and employee respondents reported eye burning, nose irritation, difficulty breathing, and vomiting. On August 11, 2015, the health department requested a health hazard evaluation by CDC's National Institute for Occupational Safety and Health to characterize the prevalence of symptoms among employees and determine the etiology of work-related symptoms. In January 2016, CDC investigators performed a cross-sectional epidemiologic study, environmental sampling, and ventilation system assessment (1). Findings suggested that chlorine disinfection byproducts and environmental conditions contributed to a higher prevalence of work-related respiratory and ocular symptoms among employees in the waterpark compared with employees in other resort areas. Recommendations included servicing the ventilation system, changing work practices to decrease the amount of disinfection byproduct precursors, and responding promptly to employee reports of symptoms.


Testing for genetic association in the presence of linkage and gene-covariate interactions.
In order to study family-based association in the presence of linkage, we extend a generalized linear mixed model proposed for genetic linkage analysis (Lebrec and van Houwelingen (2007), Human Heredity 64, 5-15) by adding a genotypic effect to the mean. The corresponding score test is a weighted family-based association tests statistic, where the weight depends on the linkage effect and on other genetic and shared environmental effects. For testing of genetic association in the presence of gene-covariate interaction, we propose a linear regression method where the family-specific score statistic is regressed on family-specific covariates. Both statistics are straightforward to compute. Simulation results show that adjusting the weight for the within-family variance structure may be a powerful approach in the presence of environmental effects. The test statistic for genetic association in the presence of gene-covariate interaction improved the power for detecting association. For illustration, we analyze the rheumatoid arthritis data from GAW15. Adjusting for smoking and anti-cyclic citrullinated peptide increased the significance of the association with the DR locus.


Effects of iso-osmolal intravenous fluid therapy on post-ischemic brain water content in the rat.
This study evaluated the effects of different iso-osmolal solutions used for intravascular volume replacement on post-ischemic cerebral edema. Fasted Sprague-Dawley rats underwent 10 min of severe (near-complete) forebrain ischemia (bilateral carotid artery occlusion and hemorrhagic hypotension). At the completion of ischemia, 40% of the estimated total blood volume was replaced by iso-osmolal saline, 6% hetastarch, or blood. Plasma osmolality remained similar between groups throughout the experiment, while saline infusion resulted in a significant but transient decrease in colloid oncotic pressure. At 1.5 h, 6 h, and 24 h post-ischemia (PI), specific gravity was determined for tissue samples taken from the hippocampus, caudoputamen, and neocortex. Normal values were obtained from rats receiving anesthesia only. The ischemic insult resulted in a significant increase in regional water content at 1.5 h PI in animals receiving blood. These values were not different from rats receiving saline or hetastarch. At 6 h PI, partial resolution of the edema was observed, with no differences in regional specific gravity occurring between fluid groups. At 24 h PI, again, no difference between fluid regimens was seen in the hippocampus or neocortex. However, in the caudoputamen, hetastarch produced a significant increase in water content relative to both saline and blood. With that exception, the authors' results indicate that early post-ischemic cerebral edema remains generally independent of iso-osmolal fluids used for resuscitation in this model of global ischemia.


Interactions of Pannexin1 channels with purinergic and NMDA receptor channels.
Pannexins are a three-member family of vertebrate plasma membrane spanning molecules that have homology to the invertebrate gap junction forming proteins, the innexins. However, pannexins do not form gap junctions but operate as plasma membrane channels. The best-characterized member of these proteins, Pannexin1 (Panx1) was suggested to be functionally associated with purinergic P2X and N-methyl-D-aspartate (NMDA) receptor channels. Activation of these receptor channels by their endogenous ligands leads to cross-activation of Panx1 channels. This in turn potentiates P2X and NMDA receptor channel signaling. Two potentiation concepts have been suggested: enhancement of the current responses and/or sustained receptor channel activation by ATP released through Panx1 pore and adenosine generated by ectonucleotidase-dependent dephosphorylation of ATP. Here we summarize the current knowledge and hypotheses about interactions of Panx1 channels with P2X and NMDA receptor channels. This article is part of a Special Issue entitled: Gap Junction Proteins edited by Jean Claude Herve.


Effect of amiodarone on Kv1.4 channel C-type inactivation: comparison of its effects with those induced by propafenone and verapamil.
As the major component of I(to) (slow), Kv1.4 channel plays an important role in repolarization of cardiac myocytes. C-type inactivation is one of Kv1.4 inactivation and can be affected by open channel blockers. We used the two-electrode voltage clamp technique to observe the effect of amiodarone on Kv1.4 C-type inactivation and compare amiodarone's effects on Kv1.4 with propafenone and verapamil. Our data show that those three antiarrhythmic drugs blocked fKv1.4 delta N (N-terminal deleted Kv1.4 channel from ferret heart) in voltage- and frequent-dependent manners. The amiodarone's IC50 was 489.23 +/- 4.72 microM, higher than that of propafenone (98.97 +/- 1.13 microM) and verapamil (263.26 +/- 6.89 microM) for fKv1.4 delta N channel (+50 mV). After application of amiodarone, propafenone and verapamil, fKv1.4 delta N inactivation becomes bi-exponential: the faster portion of inactivation (drug-induced inactivation) and the slower portion of inactivation (C-type inactivation). Amiodarone and verapamil fastened C-type inactivation in fKv1.4 delta N, but propafenone did not. Unlike propafenone that had no effect on fKv1.4 delta N recovery, amiodarone and verapamil slowed recovery in fKv1.4 delta N.


Thermal properties of lipid bilayers derived from the transient heating regime of upconverting nanoparticles.
Heat transfer and thermal properties at the nanoscale can be challenging to obtain experimentally. These are potentially relevant for understanding thermoregulation in cells. Experimental data from the transient heating regime in conjunction with a model based on the energy conservation enable the determination of the specific heat capacities for all components of a nanoconstruct, namely an upconverting nanoparticle and its conformal lipid bilayer coating. This approach benefits from a very simple, cost-effective and non-invasive optical setup to measure the thermal parameters at the nanoscale. The time-dependent model developed herein lays the foundation to describe the dynamics of heat transfer at the nanoscale and were used to understand the heat dissipation by lipid bilayers.


Linking the timing of a mother's and child's death: Comparative evidence from two rural South African population-based surveillance studies, 2000-2015.
The effect of the period before a mother's death on child survival has been assessed in only a few studies. We conducted a comparative investigation of the effect of the timing of a mother's death on child survival up to age five years in rural South Africa. We used discrete time survival analysis on data from two HIV-endemic population surveillance sites (2000-2015) to estimate a child's risk of dying before and after their mother's death. We tested if this relationship varied between sites and by availability of antiretroviral therapy (ART). We assessed if related adults in the household altered the effect of a mother's death on child survival. 3,618 children died from 2000-2015. The probability of a child dying began to increase in the 7-11 months prior to the mother's death and increased markedly in the 3 months before (2000-2003 relative risk = 22.2, 95% CI = 14.2-34.6) and 3 months following her death (2000-2003 RR = 20.1; CI = 10.3-39.4). This increased risk pattern was evident at both sites. The pattern attenuated with ART availability but remained even with availability at both sites. The father and maternal grandmother in the household lowered children's mortality risk independent of the association between timing of mother and child mortality. The persistence of elevated mortality risk both before and after the mother's death for children of different ages suggests that absence of maternal care and abrupt breastfeeding cessation might be crucial risk factors. Formative research is needed to understand the circumstances for children when a mother is very ill or dies, and behavioral and other risk factors that increase both the mother and child's risk of dying. Identifying families when a mother is very ill and implementing training and support strategies for other members of the household are urgently needed to reduce preventable child mortality.


Standardization of immunoglobulin M capture enzyme-linked immunosorbent assays for routine diagnosis of arboviral infections.
Immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA) is a rapid and versatile diagnostic method that readily permits the combination of multiple assays. Test consolidation is especially important for arthropod-borne viruses (arboviruses) which belong to at least three virus families: the Togaviridae, Flaviviridae, and Bunyaviridae. Using prototype viruses from each of these families and a panel of well-characterized human sera, we have evaluated and standardized a combined MAC-ELISA capable of identifying virus infections caused by members of each virus family. Furthermore, by grouping antigens geographically and utilizing known serological cross-reactivities, we have reduced the number of antigens necessary for testing, while maintaining adequate detection sensitivity. We have determined that a 1:400 serum dilution is most appropriate for screening antiviral antibody, using a positive-to-negative ratio of >/=2.0 as a positive cutoff value. With a blind-coded human serum panel, this combined MAC-ELISA was shown to have test sensitivity and specificity that correlated well with those of other serological techniques.


Flavonoid phytochemicals regulate activator protein-1 signal transduction pathways in endometrial and kidney stable cell lines.
Phytochemicals bind to and regulate the human estrogen receptors (ERalpha and ERbeta), mimicking actions of the endogenous estrogen, 17beta-estradiol, and known antiestrogens such as ICI 182,780. Recently, however, some of these estrogenic phytochemicals have been shown to affect other signal transduction pathways, such as receptor tyrosine kinases and mitogen-activated protein kinases (MAPK). Previously, we found that certain phytochemicals, such as flavone, apigenin, kaempferide and chalcone, have potent antiestrogenic activity. However, the antiestrogenicity of these compounds does not correlate with their ER binding capacity, suggesting alternative signaling as a mechanism for their antagonistic effects. In this study, we examined the effects of these compounds on the transcription factor activator protein-1 (AP-1). Using AP-1-luciferase stable human endometrial adenocarcinoma Ishikawa and human embryonic kidney (HEK) 293 cells, chalcone, flavone and apigenin all stimulated AP-1 activity. Additionally, we determined the effects of the phytochemicals on transcription factors that are downstream targets of various MAPK pathways. To test this, we used HEK 293 cells stably cointegrated with GAL4 transcriptional activation systems of Elk-1, c-Jun or C/EBP homologous protein (CHOP). Chalcone was the only phytochemical that activated all three transcription factors [Elk-1, 2.7-fold (P < 0.001); c-Jun, 2.7-fold (P = 0.025); CHOP, 3.0-fold (P = 0.002)], whereas apigenin stimulated CHOP (3.9-fold; P < 0.001), but inhibited phorbol myristoyl acetate-induced c-Jun activity (71%;P = 0.006). This work suggests that phytochemicals affect multiple signaling pathways that converge at the level of transcriptional regulation. The ability of flavonoids to regulate MAPK-responsive pathways in a selective manner indicates a mechanism by which phytochemicals may influence human health and disease.


Resynthesis of sphingomyelin from plasma-membrane phosphatidylcholine in BHK cells treated with Staphylococcus aureus sphingomyelinase.
About 60-65% of the total sphingomyelin in intact BHK cells is in a readily accessible pool which is rapidly degraded by Staphylococcus aureus sphingomyelinase. No more sphingomyelin is broken down in cells which have been fixed with glutaraldehyde or lysed with streptolysin O, suggesting that all the sphingomyelin which is available to the enzyme is on the cell surface. The inaccessible pool of sphingomyelin does not equilibrate with the plasma-membrane pool, even after prolonged incubation. Experiments using [3H]-choline show that much more phosphocholine is released from the intact cells treated with sphingomyelinase than can be accounted for by breakdown of the original cell-surface pool of sphingomyelin; the excess appears to be a consequence of the breakdown of sphingomyelin newly resynthesized at the expense of a pool of phosphatidylcholine which represents about 8% of total cell phosphatidylcholine and may reside in the plasma membrane. This would be consistent with resynthesis of cell-surface sphingomyelin by the phosphatidylcholine: ceramide phosphocholinetransferase pathway, which has previously been shown to be localized in the plasma membrane. However, in [3H]palmitate-labelled cells there appeared to be no accumulation of the diacylglycerol expected to be produced by this reaction, and no enhanced synthesis of phosphatidate or phosphatidylinositol; instead there was an increased synthesis of triacylglycerol. A similar increase in labelling of triacylglycerol was seen in enzyme-treated cells where the sphingomyelinase was subsequently removed, allowing resynthesis of sphingomyelin which occurred at a rate of about 25% of total sphingomyelin/h. Treatment of BHK cells with sphingomyelinase caused no change in the rates of fluid-phase endocytosis or exocytosis as measured with [3H]inulin.


Electrophysiological and pathological studies in spinocerebellar degenerations.
Electrophysiological and pathological studies have been performed on the peripheral nerves of patients with spinocerebellar degenerations. In Friedreich's ataxia there is impairment of sensory conduction and mild slowing of motor conduction which may be correlated with loss of large diameter fibres in the sural nerve. The abnormalities in Friedreich's ataxia are present early, in the first decade. In hereditary spastic paraparesis the nerve conduction studies and sural nerve biopsy are normal. In the other spinocerebellar degeneration and a mild loss of myelinated fibres in the sural nerve. However the abnormalities differ from those seen in Friedreich's ataxia.


Incidental adreno-cortical adenoma, why surgery? a case report.
Incidental adrenal tumors are commonly benign, but reports demonstrate that if the characteristics of the tumor are not clear, on images surgery is the procedure of choice. Our objective through this case is to show that laparoscopic adrenalectomy is a safe approach for adrenal incidental tumor regardless of radiological findings. A 52 year-old female with arterial hypertension (HBP), cramps, and back pain at right side, probably related to her chronic back pain history. She went for check up and a left adrenal mass on MRI described as myelolipoma was found incidentally. Laboratories were unremarkable except aldosterone and cortisol levels were slightly elevated. She was treated for hypertension for about a year. The patient underwent to a successful laparoscopic left adrenalectomy, after which the patient blood pressure was stable with out any medications, and the aldosterone and cortisol levels decreased. The pathological report was adrenal cortical adenoma with central hemorrhage and not a myelolipoma as described in images on magnetic resonance imaging (MRI). The use of imaging for diagnosis, clinical management and decision making is very controversial. Laparoscopic surgery for adrenal masses is a safe procedure for tumors of 6 cm regardless of the radiological description.


WNT5A expression increases during melanoma progression and correlates with outcome.
Wnt ligands play a major role in development and are important in cancer. Expression microarray analysis correlates one member of this family, WNT5A, to a subclass of melanomas with increased motility and invasion. There are no large studies of clinical samples primarily addressing the importance of WNT5A in melanoma progression or outcome. Therefore, this study aimed to assess the protein expression of WNT5A during melanoma progression and its effect on outcome. Expression of WNT5A was determined in a series of 59 primary melanomas with matched metastases. To provide a benchmark of progression against which to assess WNT5A, expression of p16(ink4a) was analyzed, as this has been previously well documented in melanoma. The effect of WNT5A protein expression on outcome was assessed in 102 melanomas. Cytoplasmic WNT5A showed a trend of increasing expression with melanoma progression (P = 0.013), whereas there was diminishing p16(ink4a) expression (P = 0.006). Nevi showed relatively strong WNT5A expression. Strong cytoplasmic WNT5A was an independent risk factor for reduced metastasis-free and overall survival in multivariate analysis (P = 0.001 and 0.003, respectively). Cytoplasmic WNT5A increases with melanoma progression and strong expression is associated with poor outcome.


Sagittal plane walking patterns are related to MRI changes over 18-months in people with and without mild-moderate hip osteoarthritis.
The purpose was to evaluate the association of sagittal plane gait mechanics with MRI changes in the hip joint over 18-months. Subjects with and without radiographic hip OA (n = 57) underwent MRI at baseline and 18 months for grading of cartilage lesions, bone marrow lesions (BML), cysts, and labral tears. 3D gait analyses at baseline were used for sagittal plane hip kinematics and kinetics during the stance phase. Subjects were classified as progressors or non-progressors based on increase in any MRI OA parameter. Multivariate ANOVA were used for differences in sagittal gait parameters between progressors and non-progressors at baseline while adjusting for age. Logistic regression was used to estimate the probability of being classified as a progressor or non-progressor with increasing hip flexion while adjusting for age, BMI, sex, and presence of radiographic hip OA. Of the 57, 35 were classified as non-progressors and 22 were classified as progressors. At baseline, the progressors walked with 4.5° greater hip flexion during early stance (p = 0.021) and 3.5° lesser hip extension in late stance that was nearly significant (p = 0.059). Walking with greater hip flexion at baseline was associated with a greater risk of increase in MRI defined structural changes in the hip joint (Odds Ratio = 1.1, p = 0.038). Greater hip flexion during walking was associated with a risk of structural progression of hip OA. The results may guide future interventions to alter the walking patterns and slow structural hip OA progression.© 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1472-1477, 2018.


New colleges of osteopathic medicine: steps in achieving accreditation.
The authors outline the process that an educational institution undergoes to obtain accreditation status from the American Osteopathic Association's Commission on Osteopathic College Accreditation (COCA), the body responsible for accrediting the nation's osteopathic medical schools. Progress toward accreditation status is described for the six newest colleges of osteopathic medicine (COMs). In addition, the authors describe how the expansion of a COM's approved class size-whether as a result of planned growth or methodologic changes in educational programming-is considered by US federal administrative regulations a "substantive change" to the institution's mission and programs. Such changes require the review and approval of COCA. Class-size increases for the past decade are detailed.