Most efficient routes for the synthesis of alpha,beta-diamino acid-derived compounds.
Alpha,beta-Diamino acids have attracted considerable attention recently due to their growing importance in pharmaceutical and biochemical research. For example, this special class of alpha,beta-diamino acids has become the components of enzyme inhibitors, and has been incorporated into peptides which are used to modulate secondary and tertiary structural conformations. Although their widely occurrence in nature, optically active diamino acids are hard to isolate and purify from available natural resources on large scale. Therefore, their asymmetric synthesis becomes a great interest for organic and medicinal chemists. However, there still exist great challenges for enantioselective synthesis of diamino acids, especially those with two vicinal chiral centers. This review highlights the recent promising methodologies for enantioselective synthesis of alpha,beta-diamino acids, with special emphasis on catalytic asymmetric reactions, as well as methods for natural chiral compound derivatization, and chiral auxiliaries.


Chloroquine resistance produced in vitro in an African strain of human malaria.
After condinous cultivation in the presence of chloroquine, an African strain of the malaria parasite, Plasmodium falciparu, acquired resistance to the drug. The resistance was stable and comparable in vitro to that occurring naturally in a strain from Southeast Asia. This suggests that chloroquine resistance, absent until now in Africa, might arise in the future.


[Functional morphology of the neonatal umbilical cord during cesarean section].
The morphology of the neonatal umbilical cord was studied in 23 women during elective surgical delivery and in 32 women during normal physiological delivery. Organ- and histometric indices were used. These included: length, weight, a linear weight unit; the average diameter of the umbilical cord, etc. The significant increase of organometric indices (a linear weight unit of weight, the average diameter of the umbilical cord) and histometric ones (a drastic venous lumen increase, a significant increase in the proportion of Wharton's jelly) document acute umbilical blood flow disorder in the absence of the conditions characteristic for normal delivery. Moderately pronounced edema of the umbilical cord peripheral layer emerges. A complex of morphofunctional umbilical cord changes during surgical delivery serves as predictor of neonatal respiratory distress syndrome.


A comparison of single-versus double-row suture anchor techniques in a simulated repair of the rotator cuff: an experimental study in rabbits.
We compared time-dependent changes in the biomechanical properties of single-and double-row repair of a simulated acute tear of the rotator cuff in rabbits to determine the effect of the fixation techniques on the healing process. A tear of the supraspinatus tendon was created in 80 rabbits which were separated into two equal groups. A single-row repair with two suture anchors was conducted in group 1 and a double-row repair with four suture anchors in group 2. A total of ten intact contralateral shoulder joints was used as a control group. Biomechanical testing was performed immediately post-operatively and at four and eight weeks, and histological analysis at four and eight weeks. The mean load to failure in group 2 animals was greater than in group 1, but both groups remained lower than the control group at all intervals. Histological analysis showed similar healing properties at four and eight weeks in both groups, but a significantly larger number of healed tendon-bone interfaces were identified in group 2 than in group 1 at eight weeks (p < 0.012). The ultimate load to failure increased with the number of suture anchors used immediately post-operatively, and at four and eight weeks. The increased load to failure at eight weeks seemed to be related to the increase in the surface area of healed tendon-to-bone in the double-row repair group.


Kaposi's Sarcoma-Associated Herpesvirus ORF50 Protein Represses Cellular MDM2 Expression via Suppressing the Sp1- and p53-Mediated Transactivation.
The Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded ORF50 protein is a potent transcriptional activator essential for triggering KSHV lytic reactivation. Despite extensive studies, little is known about whether ORF50 possesses the ability to repress gene expression or has an antagonistic action to cellular transcription factors. Previously, we demonstrated that human oncoprotein MDM2 can promote the degradation of ORF50 protein. Herein, we show that abundant ORF50 expression in cells can conversely downregulate MDM2 expression via repressing both the upstream (P1) and internal (P2) promoters of the MDM2 gene. Deletion analysis of the MDM2 P1 promoter revealed that there were two ORF50-dependent negative response elements located from -102 to -63 and from -39 to +1, which contain Sp1-binding sites. For the MDM2 P2 promoter, the ORF50-dependent negative response element was identified in the region from -110 to -25, which is coincident with the location of two known p53-binding sites. Importantly, we further demonstrated that overexpression of Sp1 or p53 in cells indeed upregulated MDM2 expression; however, coexpression with ORF50 protein remarkably reduced the Sp1- or p53-mediated MDM2 upregulation. Collectively, our findings propose a reciprocal negative regulation between ORF50 and MDM2 and uncover that ORF50 decreases MDM2 expression through repressing Sp1- and p53-mediated transactivation.


Pressure dissociation of integration host factor-DNA complexes reveals flexibility-dependent structural variation at the protein-DNA interface.
E. coli Integration host factor (IHF) condenses the bacterial nucleoid by wrapping DNA. Previously, we showed that DNA flexibility compensates for structural characteristics of the four consensus recognition elements associated with specific binding (Aeling et al., J. Biol. Chem. 281, 39236-39248, 2006). If elements are missing, high-affinity binding occurs only if DNA deformation energy is low. In contrast, if all elements are present, net binding energy is unaffected by deformation energy. We tested two hypotheses for this observation: in complexes containing all elements, (1) stiff DNA sequences are less bent upon binding IHF than flexible ones; or (2) DNA sequences with differing flexibility have interactions with IHF that compensate for unfavorable deformation energy. Time-resolved Förster resonance energy transfer (FRET) shows that global topologies are indistinguishable for three complexes with oligonucleotides of different flexibility. However, pressure perturbation shows that the volume change upon binding is smaller with increasing flexibility. We interpret these results in the context of Record and coworker's model for IHF binding (J. Mol. Biol. 310, 379-401, 2001). We propose that the volume changes reflect differences in hydration that arise from structural variation at IHF-DNA interfaces while the resulting energetic compensation maintains the same net binding energy.


Isolation and characterization of cytoplasmic membranes and chlorosomes from the green bacterium Chloroflexus aurantiacus.
A method was developed which allows the isolation and purification of cytoplasmic membranes and chlorosomes from cells of Chloroflexus aurantiacus grown under different light conditions. The dipolar ionic detergent Deriphat (0.08%) and a sodium iodide gradient centrifugation were used in isolating cytoplasmic membranes. Chlorosomes were prepared with 0.16% of the dipolar ionic detergent Miranol and purified by a sucrose gradient centrifugation. Cytoplasmic membrane fractions prepared from either high- (3,000 W m-2), medium-(200 W m-2) or low- (7 W m-2) light-grown cells had near infrared absorption bands at 866, 808, and 755 nm in a constant characteristic absorbance ratio of 6:3.8:1. In all cytoplasmic membrane preparations, the amount of bacteriochlorophyll a (Bchl a) per cytochrome, the amount of Bchl a per reaction center, and reaction center per milligram of cytoplasmic membrane protein was found to be constant. No Bchl c was present. Five respiratory enzyme activities have been measured in the cytoplasmic membrane fraction. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of denatured cytoplasmic membrane showed many bands, but a major polypeptide with an apparent molecular weight of 8,000. In contrast, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified chlorosomes did not contain the 8,000-molecular-weight band but revealed only three distinct protein bands with molecular weights of 15,000, 12,000, and 6,000. Isolated chlorosomes contained Bchl c and a small, yet constant, amount of Bchl a (absorbing at 790 nm) in a molar ratio of 25:1. The data indicated that the components of the photosynthetic apparatus in the cytoplasmic membrane of Chloroflexus aurantiacus remained constant and only the amount of antenna Bchl c varied with light conditions.


The Use of Telemedicine in Cancer Clinical Trials: Connect-Patient-to-Doctor Prospective Study.
Telemedicine is currently being adopted for the management of patients in routine care. However, its use remains limited in the context of clinical trials. This study aimed to demonstrate the feasibility of telemonitoring and patient-reported outcomes collection in the context of clinical trials. The patients who were included in an interventional oncology clinical trial were eligible. The patients were registered with a digital tool to respond to a patient-reported outcomes questionnaire (ePRO) based on CTCAE (The Common Terminology Criteria for Adverse Events, National Cancer Institute), version 5.0, personalized to their pathology and treatment. An algorithm evaluated the health status of the patient based on the reported adverse events, with a classification in 4 different states (correct, compromise, state to be monitored, or critical state). The main objective was to evaluate the feasibility of remote monitoring via a connected platform of patients included in a clinical trial. From July 1, 2020, to March 31, 2021, 39 patients were included. The median age was 71 years (range 41-94); 74% (n=29) were male, and 59% (n=23) had metastatic disease. Out of the 969 ePRO questionnaires completed over the course of the study, 77.0% (n=746) were classified as "correct," 10.9% (n=106) as "compromised," and 12.1% (n=117) as "to be monitored" or "critical." The median response time was 7 days (IQR 7-15.5), and 76% (25/33) of the patients were compliant. Out of the 35 patients who answered a satisfaction questionnaire, 95% (n=33) were satisfied or very satisfied with the tool, and 85% (n=30) were satisfied with their relationship with the health care team. There were 5 unscheduled hospitalizations during the study period. Remote monitoring in clinical trials is feasible, with a high level of patient participation and satisfaction. It benefits patients, but it also ensures the high quality of the trial through the early management of adverse events and better knowledge of the tolerance profile of experimental treatments. This e-technology will likely be deployed more widely in our clinical trials.


Responsibility for retirement planning shifts to employees.
In recent years, organizations have shifted away from offering their employees defined benefit plans that promise specified income streams to employees when they retire. Instead, they are offering employees defined contribution plans, which rely on investment performance, as directed by plan participants, to generate sufficient retirement income. Healthcare financial managers who work in organizations that offer retirement plans for their employees find themselves increasingly prevailed upon to play the role of instructor to plan participants. While some financial managers may not relish the role, the more successful financial managers are at helping plan participants achieve their retirement income goals, the better the chance of healthcare organizations avoiding future liability problems.


Reflective, causal, and composite indicators of quality of life: A conceptual or an empirical distinction?
Items (or indicators) that constitute "quality of life" instruments can be classified as either reflective (manifestations of some underlying construct), causal (the construct is an effect of the indicators), or composite (the construct is an exact linear combination of the indicators). Psychometric methods based on inter-item associations are only appropriate for reflective indicators, whereas other statistical and non-statistical validation methods can be used for composite or causal indicators. Thus, the distinction has important practical, as well as theoretical, implications. Attempts have been made to empirically identify which items of the EORTC QLQ-C30, a cancer-specific instrument, are causal and which are reflective. Such attempts, however, first require commitment to a particular definition of quality of life, of which there are many. Whether an indicator forms a composite, is causal or reflective of quality of life will depend on the definition adopted, and therefore, the reflective-composite-causal distinction is, arguably, best established on conceptual rather empirical grounds, guided by the "mental experiments" suggested by Bollen (Structural equations with latent variables, Wiley, New York, 1989). Conceptual models of health status and quality of life, as well as a cognitive-linguistic approach to quality of life assessment, may make some contribution to this practice. Theoretical consideration of indicator content can guide not only instrument development and validation, but also the selection of an appropriate instrument.


Distributed multiple description video coding on packet loss channels.
In this paper, we are to solve the drift problem of multiple description video coding on packet loss channels by using state-of-the-art distributed techniques. We first present an asymptotically optimal code design of multiple descriptions in the Wyner-Ziv (MDWZ) setting. Then we propose a distributed multiple description video coding (DMDVC) scheme, which performs MDWZ coding on each nonintra coded frame. Instead of the prediction loops used in traditional multiple description video coding, Slepian-Wolf based coding is used to exploit interframe correlations. A bitplane extraction scheme is proposed to improve the balance between two descriptions, so that side informations can be interchanged between the side decoders of DMDVC with negligible quality degradation, which is crucial to robust transmission over packet loss channels. Experiment results demonstrate the robustness of our scheme, especially at high packet loss rates.


Preclinical Evidence and Possible Mechanisms of Extracts or Compounds from Cistanches for Alzheimer's Disease.
Currently, disease-modified strategies to prevent, halt or reverse the progress of Alzheimer's disease (AD) are still lacking. Previous studies indicated extracts or compounds from Cistanches (ECC) exert a potential neuroprotective effect against AD. Thus, we conducted a preclinical systematic review to assess preclinical evidence and possible mechanisms of ECC in experimental AD. A systematical searching strategy was carried out across seven databases from their inceptions to July 2018. Twenty studies with 1696 rats or mice were involved. Neurobehavioral function indices as primary outcome measures were established by the Morris water maze test (n = 11), step-down test (n = 10), electrical Y-maze test (n = 4), step-through test (n = 3), open field test (n = 2) and passage water maze test (n = 1). Compared with controls, the results of the meta-analysis showed ECC exerted a significant effect in decreasing the escape latency, error times and wrong reaction latency in both the training test and the retention test, and in increasing the exact time and the percentage of time in the platform-quadrant and the number of platform crossings (all P<0.01). In conclusion, ECC exert potential neuroprotective effects in experimental AD, mainly through mechanisms involving antioxidant stress and antiapoptosic effects, inhibiting Aβ deposition and tau protein hyperphosphorylation and promoting synapse protection. Thus, ECC could be a candidate for AD treatment and further clinical trials.


Effects of age and body proportions on stature estimation.
Two issues involved in mathematical estimation of stature from long bone lengths are explored: the use of different age points for estimating maximum adult stature, and the effects of linear body proportions on stature estimation errors. Both issues were raised by a recent analysis of stature in the British Medieval Wharram Percy sample. A large (n > 500) sample of European skeletal remains with anatomically estimated statures is used to test associations between relative lower limb length and errors in stature estimation using previously published equations for European samples. Two cadaveric samples of known ages (Terry and Bass Collections) are used to identify the most appropriate age point to employ in a linear equation with an age term for estimation of maximum adult anatomical stature. Relative lower limb length is positively correlated with errors in stature estimation from lower limb bone lengths. Underestimation of stature in the Wharram Percy sample by the European equations is largely attributable to the relatively short lower limbs of this sample compared to Europeans in general. Two methods for assessing and adjusting for relative lower limb length variation are presented. Maximum adult stature is best estimated using an age point of 30 years when a linear age term is employed. Body proportions may vary even within relatively closely related populations, so should be assessed and compared to those of reference samples whenever possible when applying mathematical stature equations.


The Kinematic-Muscle Synergies during Infant Crawling: A Pilot Study.
Many feature extraction algorithms have been separately used for kinematic or muscle synergy analysis during human movement. However, very few studies focus on the co-extraction of kinematic and muscle synergies. Therefore, the aim of this study was to propose a novel and efficient approach for extracting the kinematic-muscle synergies during infant crawling. Surface electromyography signals and three-dimensional joint trajectories were collected from 20 typically developing infants during self-paced hands-and-knees crawling. Angular accelerations of shoulder, elbow, hip and knee flexion/extension computing from those joint trajectories were divided into two independent directional positive degrees-of-freedom. The kinematic-muscle synergies and corresponding activation coefficients were extracted using the non-negative matrix factorization algorithm based on two selection criteria of synergy number (i.e., criterion 1: the total constraint, criterion 2: a combination of the total constraint and a local constraint for each joint/muscle). Then, the data of each joint/muscle were reconstructed by those synergies and corresponding activation coefficients. Our results indicated that the minimum number of kinematic-muscle synergies based on criterion 1 is less than that based on criterion 2. The data reconstruction of joint flexion/extension based on criterion 2 is better than that based on criterion 1, whereas the data reconstruction of muscles is similar between criterion 1 and 2. These promising results show the feasibility of applying the proposed approach to clinical assessments of motor function for infants.Clinical Relevance- Extracting kinematic-muscle synergies during infant crawling has the potential for professional therapists or rehabilitation physicians to conduct the early assessment and rehabilitation treatment of infants with the central nervous system disorders.


[Elementary neuronal ensembles as the basis for the functional organization of the brain].
The structural basis of neuronal assemblies is made up of morphological associations of neurones determined by the properties of the central projection of afferent pathways. Underlying the functional organization of probabilistic neuronal assemblies is the formation of conjugated relations between neurones of the central zone of synergically activated units and the surrounding zone of inhibited neurones. The characteristics of informational activity and neuronal assemblies are related to the optimization of perception processes and reorganization of sensory information, and to the selection of useful information from the general sensory flow, especially in the course of formation of a habit. The observed characteristics of the work of probabilistic neuronal assemblies are considered as manifestations of formation of structural, functional and informational redundancy, the basic mechanism ensuring reliable functioning of the informational-controlling brain systems.


Wheelchair users are not necessarily wheelchair bound.
To determine the patterns of wheelchair use in terms of locations of use, whether wheelchair use in one location was related to wheelchair use in other locations, and factors associated with wheelchair use in different locations. Longitudinal cohort study. Patients prescribed wheelchairs by clinicians at one of two teaching hospitals (one Veterans Affairs hospital and one private hospital). One hundred fifty-three consecutive persons who were prescribed a new wheelchair, resided in the community, had a Short Portable Mental Status Questionnaire score of greater than six out of 10, and who could be interviewed within 7 to 21 days of receiving the wheelchair. Patient, wheelchair, and environmental characteristics and self-reported wheelchair use in life spaces. Wheelchair use in the 24 hours before the interview was inconsistent across life spaces. The correlation between wheelchair use in the bath and in the kitchen was 0.66, between locations near and far from home was -0.08, and between locations in the home and outside the home was 0.08. Predictors of wheelchair use in the home were using help from another person to propel the wheelchair (odds ratio (OR) = 0.14, 95% confidence interval (CI) = 0.04-0.45), the number of impairments (OR = 0.80, 95% CI = 0.67-0.96), a report that the wheelchair did not meet the subject's needs (OR = 3.71, 95% CI = 1.27-10.81), and having adapted the home to accommodate the wheelchair (OR = 3.75, 95% CI = 1.47-8.18). Having adapted the home was also positively associated with use of the wheelchair in areas near the home (OR = 4.77, 95% CI = 1.94-11.71). The only factor associated with wheelchair use in distant locations was older age (OR = 0.62, 95% CI = 0.46-0.83 per 10-year increment). Personal factors (e.g., using help to propel the wheelchair) and environmental factors (e.g., home adaptations to accommodate the wheelchair) influenced wheelchair use. In addition, wheelchair use, and the factors influencing wheelchair use, differed by location. Wheelchair users appear to use their wheelchairs selectively, depending on their physical needs and the constraints of their environment.


Colistin- and Carbapenem-Resistant Escherichia coli Harboring mcr-1 and blaNDM-5, Causing a Complicated Urinary Tract Infection in a Patient from the United States.
Colistin is increasingly used as an antibiotic of last resort for the treatment of carbapenem-resistant Gram-negative infections. The plasmid-borne colistin resistance gene mcr-1 was initially identified in animal and clinical samples from China and subsequently reported worldwide, including in the United States. Of particular concern is the spread of mcr-1 into carbapenem-resistant bacteria, thereby creating strains that approach pan-resistance. While several reports of mcr-1 have involved carbapenem-resistant strains, no such isolates have been described in the United States. Here, we report the isolation and identification of an Escherichia coli strain harboring both mcr-1 and carbapenemase gene blaNDM-5 from a urine sample in a patient without recent travel outside the United States. The isolate exhibited resistance to both colistin and carbapenems, but was susceptible to amikacin, aztreonam, gentamicin, nitrofurantoin, tigecycline, and trimethoprim-sulfamethoxazole. The mcr-1- and blaNDM-5-harboring plasmids were completely sequenced and shown to be highly similar to plasmids previously reported from China. The strain in this report was first isolated in August 2014, highlighting an earlier presence of mcr-1 within the United States than previously recognized. Colistin has become the last line of defense for the treatment of infections caused by Gram-negative bacteria resistant to multiple classes of antibiotics, in particular carbapenem-resistant Enterobacteriaceae (CRE). Resistance to colistin, encoded by the plasmid-borne gene mcr-1, was first identified in animal and clinical samples from China in November 2015 and has subsequently been reported from numerous other countries. In April 2016, mcr-1 was identified in a carbapenem-susceptible Escherichia coli strain from a clinical sample in the United States, followed by a second report from a carbapenem-susceptible E. coli strain originally isolated in May 2015. We report the isolation and identification of an E. coli strain harboring both colistin (mcr-1) and carbapenem (blaNDM-5) resistance genes, originally isolated in August 2014 from urine of a patient with recurrent urinary tract infections. To our knowledge, this is the first report in the United States of a clinical bacterial isolate with both colistin and carbapenem resistance, highlighting the importance of active surveillance efforts for colistin- and carbapenem-resistant organisms.


Primary afferent plasticity following deafferentation of the trigeminal brainstem nuclei in the adult rat.
Alpha-tyrosinated tubulin is a cytoskeletal protein that is involved in axonal growth and is considered a marker of neuronal plasticity in adult mammals. In adult rats, unilateral ablation of the left facial sensorimotor cortical areas induces degeneration of corticotrigeminal projections and marked denervation of the contralateral sensory trigeminal nuclei. Western blotting and real-time-PCR of homogenates of the contralateral trigeminal ganglion (TG) revealed consistent overexpression of growth proteins 15 days after left decortication in comparison with the ipsilateral side. Immunohistochemical analyses indicated marked overexpression of alpha-tyrosinated tubulin in the cells of the ganglion on the right side. Cytoskeletal changes were primarily observed in the small ganglionic neurons. Application of HRP-CT, WGA-HRP, and HRP to infraorbital nerves on both sides 15 days after left decortication showed a significant degree of terminal sprouting and neosynaptogenesis from right primary afferents at the level of the right caudalis and interpolaris trigeminal subnuclei. These observations suggest that the adaptive response of TG neurons to central deafferentation, leading to overcrowding and rearrangement of the trigeminal primary afferent terminals on V spinal subnuclei neurons, could represent the anatomical basis for distortion of facial modalities, perceived as allodynia and hyperalgesia, despite nerve integrity.


Cardiac depression induced by cocaine or cocaethylene is alleviated by lipid emulsion more effectively than by sulfobutylether-β-cyclodextrin.
Cocaine intoxication leads to over 500,000 emergency department visits annually in the United States and ethanol cointoxication occurs in 34% of those cases. Cardiotoxicity is an ominous complication of cocaine and cocaethylene overdose for which no specific antidote exists. Because infusion of lipid emulsion (Intralipid) can treat lipophilic local anesthetic toxicity and cocaine is an amphipathic local anesthetic, the authors tested whether lipid emulsion could attenuate cocaine cardiotoxicity in vivo. The effects of lipid emulsion were compared with the metabolically inert sulfobutylether-β-cyclodextrin (SBE-β-CD; Captisol) in an isolated heart model of cocaine and cocaethylene toxicity to determine if capture alone could exert similar benefit as lipid emulsion, which exhibits multimodal effects. The authors then tested if cocaine and cocaethylene, like bupivacaine, inhibit lipid-based metabolism in isolated cardiac mitochondria. For whole animal experiments, Sprague-Dawley rats were anesthetized, instrumented, and pretreated with lipid emulsion followed by a continuous infusion of cocaine to assess time of onset of cocaine toxicity. For ex vivo experiments, rat hearts were placed onto a nonrecirculating Langendorff system perfused with Krebs-Henseleit solution. Heart rate, left ventricle maximum developed pressure (LVdevP), left ventricle diastolic pressure, maximum rate of contraction (+dP/dtmax), maximum rate of relaxation (-dP/dtmax), rate-pressure product (RPP = heart rate × LVdevP), and line pressure were monitored continuously during the experiment. A dose response to cocaine (10, 30, 50, and 100 μmol/L) and cocaethylene (10, 30, and 50 μmol/L) was generated in the absence or presence of either 0.25% lipid emulsion or SBE-β-CD. Substrate-specific rates of oxygen consumption were measured in interfibrillar cardiac mitochondria in the presence of cocaine, cocaethylene, ecgonine, and benzoylecgonine. Treatment with lipid emulsion delayed onset of hypotension (140 seconds vs. 279 seconds; p = 0.008) and asystole (369 seconds vs. 607 seconds; p = 0.02) in whole animals. Cocaine and cocaethylene induced dose-dependent decreases in RPP, +dP/dtmax, and -dP/dtmaxabs (p < 0.0001) in Langendorff hearts; line pressure was increased by cocaine and cocaethylene infusion, but not altered by treatment. Lipid emulsion attenuated cocaine- and cocaethylene-induced cardiac depression. SBE-β-CD alone evoked a mild cardiodepressant effect (p < 0.0001) but attenuated further cocaine- and cocaethylene-induced decrements in cardiac contractility at high concentrations of drug (100 μmol/L; p < 0.001). Finally, both cocaine and cocaethylene, but not ecgonine and benzoylecgonine, inhibited lipid-dependent mitochondrial respiration by blocking carnitine exchange (p < 0.05). A commercially available lipid emulsion was able to delay progression of cocaine cardiac toxicity in vivo. Further, it improved acute cocaine- and cocaethylene-induced cardiac toxicity in rat isolated heart while SBE-β-CD was effective only at the highest cocaine concentration. Further, both cocaine and cocaethylene inhibited lipid-dependent mitochondrial respiration. Collectively, this suggests that scavenging-independent effects of lipid emulsion may contribute to reversal of acute cocaine and cocaethylene cardiotoxicity, and the beneficial effects may involve mitochondrial lipid processing.


Sustained effects of gene-activated matrices after CNS injury.
We show that when gene-activated matrices (GAM) are placed between the proximal and distal stumps of severed rat optic nerves, DNA is retained within the GAM, promoting sustained transgene expression in the optic nerve, in the GAM itself, and, more importantly, in axotomized retinal ganglion cells (RGC). Plasmids that encode basic fibroblast growth factor (FGF2), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT3) promote sustained survival of RGC for over 3 months after the initial injury. These findings suggest that immobilized DNA implanted into a CNS lesion will be delivered by axon terminal uptake and retrograde transport to axotomized neurons. GAM may therefore be a useful agent for promoting sustained neuron survival and axon regeneration. Whether further optimization of the matrices, plasmids, promoters, and genes present in the GAM will promote even more survival or, alternatively, axon regeneration remains to be determined.


Hypnosis, attention, and time cognition.
This study examined the influence of hypnotic susceptibility (high vs. low), hypnotic condition (hypnotic vs. nonhypnotic), and attentional demands (high vs. low) on the verbal estimates (N = 510 undergraduates) of 30- and 60-second time intervals under both prospective (aware of timing) and retrospective (unaware of timing) paradigms. It was expected that prospective judgments would be greater than retrospective judgments across all factors. Consistent with existing literature, strong support was found for this hypothesis. Hypotheses generated within the trait-state paradigm were not supported. No effects were found for hypnotic susceptibility in any form. Partial support was found for the sociocognitive model. The hypnotic context, independent of level of susceptibility, apparently utilizes the attentional resources of participants and as a by-product leads to underestimation of time. Longer time intervals (60 s) were also underestimated in comparison to shorter intervals (30 s).


Desmin-related cardiomyopathy in transgenic mice: a cardiac amyloidosis.
An R120G missense mutation in the small heat shock protein alpha-B-crystallin (CryAB(R120G)) causes desmin-related cardiomyopathy (DRM). DRM is characterized by the formation of aggregates containing CryAB and desmin, and it can be recapitulated in transgenic mice by cardiac-specific expression of the mutant protein. In this article, we show that expression of CryAB(R120G) leads to the formation of electron-dense bodies characteristic of the DRMs and identify these bodies as aggresomes, which are characteristic of the neurodegenerative diseases. Cardiomyocytes transfected with adenovirus containing CryAB(R120G) establish the necessity and sufficiency of CryAB(R120G) expression for aggresome formation. The commonality of these aggresomes with oligomeric protein aggregates found in the amyloid-related degenerative diseases was corroborated by the presence of high levels of amyloid oligomers that may represent a primary toxic species in the amyloid diseases. These oligomeric amyloid intermediates are present also in cardiomyocytes derived from many human dilated and hypertrophic cardiomyopathies.


Polysomnography: addressing the need for standards.
The field of sleep disorders medicine is facing enormous challenges as it strives to gain recognition from the medical profession, the public and government. One of these challenges is to ensure that limited resources for diagnosis are used wisely. The authors argue that the standards for polysomnography developed by the Canadian Sleep Society and the Canadian Thoracic Society (see pages 1673 to 1678 of this issue) will go a long way toward ensuring that this expensive resource is used prudently. In the mean-time, more research is needed to determine valid measures of the impact of sleep disorders and to establish the reliability of different diagnostic methods.


ERα expression in T lymphocytes is dispensable for estrogenic effects in bone.
Estrogen treatment has positive effects on the skeleton, and we have shown that estrogen receptor alpha (ERα) expression in cells of hematopoietic origin contributes to a normal estrogen treatment response in bone tissue. T lymphocytes are implicated in the estrogenic regulation of bone mass, but it is not known whether T lymphocytes are direct estrogen target cells. Therefore, the aim of this study was to determine the importance of ERα expression in T lymphocytes for the estrogenic regulation of the skeleton using female mice lacking ERα expression specifically in T lymphocytes (Lck-ERα-/-) and ERαflox/flox littermate (control) mice. Deletion of ERα expression in T lymphocytes did not affect bone mineral density (BMD) in sham-operated Lck-ERα-/- compared to control mice, and ovariectomy (ovx) resulted in a similar decrease in BMD in control and Lck-ERα-/- mice compared to sham-operated mice. Furthermore, estrogen treatment of ovx Lck-ERα-/- led to an increased BMD that was indistinguishable from the increase seen after estrogen treatment of ovx control mice. Detailed analysis of both the appendicular (femur) and axial (vertebrae) skeleton showed that both trabecular and cortical bone parameters responded to a similar extent regardless of the presence of ERα in T lymphocytes. In conclusion, ERα expression in T lymphocytes is dispensable for normal estrogenic regulation of bone mass in female mice.


Contemporary view of the clinical relevance of magnesium homeostasis.
Magnesium is one of the most abundant cations in the body and is essential for a wide variety of metabolically important reactions. Serum magnesium concentration is regulated by the balance between intestinal absorption and renal excretion. Hypomagnesaemia is relatively common, with an estimated prevalence in the general population ranging from 2.5 to 15%. It may result from inadequate magnesium intake, increased gastrointestinal or renal loss or redistribution from extracellular to intracellular space. Drug-induced hypomagnesaemia, particularly related to proton-pump inhibitor (PPI) therapy, is being increasingly recognized. Although most patients with hypomagnesaemia are asymptomatic, manifestations may include neuromuscular, cardiovascular and metabolic features. Due to the kidney's ability to increase fractional excretion to nearly 100% when the renal magnesium threshold is exceeded, clinically significant hypermagnesaemia is uncommon, generally occurring only in the setting of renal insufficiency and excessive magnesium intake. Symptoms include hypotension, nausea, facial flushing, ileus and flaccid muscle paralysis. In most cases, simply withdrawing exogenous magnesium is sufficient to restore normal magnesium concentrations, although occasionally administration of intravenous calcium or even dialysis may be required.


Alteration of photosynthate partitioning by high-level expression of phosphoglucomutase in tobacco chloroplasts.
Plastidial phosphoglucomutase (PGM) plays an important role in starch synthesis and degradation. Nonetheless, the impact of enhanced plastidial PGM activity on metabolism in photosynthetic tissue is yet to be elucidated. In this study, we generated transplastomic tobacco plants overproducing Arabidopsis thaliana plastidial PGM (AtptPGM) in chloroplasts and analyzed the consequent metabolic and physiological parameters in the transplastomic plants. AtptPGM accumulated in the chloroplasts to up to 16% of total soluble protein in the leaves. PGM activity in leaves increased 100-fold relative to that of wild-type plants. The transplastomic plants were phenotypically indistinguishable in their growth rates, photosynthetic activities, and starch synthesis from wild-type plants, but hexose partitioning in the light period was dramatically different. Furthermore, alteration of extracellular invertase activity was observed in the lower leaves of the transplastomic plants. These observations suggest that high-level expression of plastidial PGM alters hexose partitioning in light periods via modification of extracellular invertase activity.


Analog mean-delay method for high-speed fluorescence lifetime measurement.
We present a new high-speed lifetime measurement scheme of analog mean-delay (AMD) method which is suitable for studying dynamical time-resolved spectroscopy and high-speed fluorescence lifetime imaging microscopy (FLIM). In our lifetime measurement method, the time-domain intensity signal of a fluorescence decay is acquired as an analog waveform. And the lifetime information is extracted from the mean temporal delay of the acquired signal. Since this method does not rely on the single-photon counting technique, the signals of multiple fluorescence photons can be acquired simultaneously. The measurement speed can be increased easily by raising the fluorescence intensity without a photon-rate limit. We have investigated various characteristics of our method in lifetime accuracy and precision as well as measurement speed. It has been found that our method can provide excellent measurement performances in various aspects. We hav demonstrated a high-speed measurement with a high photon detection rate of approximately 108 photons per second with a nearly shot noise-limited photon economy. A fluorescence lifetime of 3.2 ns was accurately determined with a standard deviation of 3% from the data acquired within 17.8 micros at a rate of 56,300 lifetime determinations per second.


S-Methylation of D- and L-penicillamine by human erythrocyte membrane thiol methyltransferase.
Human red blood cell (RBC) membranes contain a thiol methyltransferase activity that catalyzes the S-methylation of 2-mercaptoethanol (2-ME). These experiments were performed to determine whether human RBC membranes contain enzymes that can catalyze the S-methylation of D- and L-penicillamine, to determine whether those enzymes are similar to the RBC membrane thiol methyltransferase that catalyzes the S-methylation of 2-ME, and to determine whether lipophilic conjugates of the S-methyl metabolites of D- and L-penicillamine are formed by RBC membranes. Human RBC membranes were able to catalyze the S-methylation of D- and L-penicillamine. The apparent Michaelis (Km) constants for D- and L-penicillamine were 7.53 and 7.27 mM, respectively. However, the Vmax value for L-penicillamine was more than 2.5 times greater than the Vmax value for D-penicillamine. D- and L-Penicillamine methyltransferases and 2-ME thiol methyltransferase were similar with respect to their subcellular distributions, inhibitor sensitivities, and thermal stabilities. In addition, when methyltransferase activities for 2-ME and for D- and L-penicillamine were measured in RBC membranes from 19 individual subjects, there were highly significant correlations among all three activities (r greater than 0.98, p less than 0.001 for all three comparisons). These observations suggest either that a single enzyme in the human RBC membrane catalyzes the S-methylation of all three compounds, or, less likely, that these reactions are catalyzed by three separate enzymes that are regulated in parallel and have similar properties. Experiments were then performed to identify the products of the penicillamine methylation reactions.(ABSTRACT TRUNCATED AT 250 WORDS)


Protective effect of dieckol against chemical hypoxia-induced cytotoxicity in primary cultured mouse hepatocytes.
Hepatic ischemic injury is a major complication arising from liver surgery, transplantation, or other ischemic diseases, and both reactive oxygen species (ROS) and pro-inflammatory mediators play the role of key mediators in hepatic ischemic injury. In this study, we examined the effect of dieckol in chemical hypoxia-induced injury in mouse hepatocytes. Cell viability was significantly decreased after treatment with cobalt chloride (CoCl2), a well-known hypoxia mimetic agent in a time- and dose-dependent manner. Pretreatment with dieckol before exposure to CoCl2 significantly attenuated the CoCl2-induced decrease of cell viability. Additionally, pretreatment with dieckol potentiated the CoCl2-induced decrease of Bcl-2 expression and attenuated the CoCl2-induced increase in the expression of Bax and caspase-3. Treatment with CoCl2 resulted in an increased intracellular ROS generation, which is inhibited by dieckol or N-acetyl cysteine (NAC, a ROS scavenger), and p38 MAPK phosphorylation, which is also blocked by dieckol or NAC. In addition, dieckol and SB203580 (p38 MAPK inhibitor) increased the CoCl2-induced decrease of Bcl-2 expression and decreased the CoCl2-induced increase of Bax and caspase-3 expressions. CoCl2-induced decrease of cell viability was attenuated by pretreatment with dieckol, NAC, and SB203580. Furthermore, dieckol attenuated CoCl2-induced COX-2 expression. Similar to the effect of dieckol, NAC also blocked CoCl2-induced COX-2 expression. Additionally, CoCl2-induced decrease of cell viability was attenuated not only by dieckol and NAC but also by NS-398 (a selective COX-2 inhibitor). In conclusion, dieckol protects primary cultured mouse hepatocytes against CoCl2-induced cell injury through inhibition of ROS-activated p38 MAPK and COX-2 pathway.


The outcomes of primary scleral buckling during repair of posterior segment open-globe injuries.
To compare visual outcomes of eyes which underwent primary scleral buckling (PSB) treatment during posterior segment open-globe injury (OGI) repair with eyes not treated with PSB. We retrospectively reviewed 38 eyes which underwent a posterior segment OGI repair with no preoperative evidence of retinal detachment (RD) at Soroka University Medical Center (1995-2010). 19 (50%) underwent scleral repair alone (control group) and the other 19 eyes were treated with PSB also (PSB group). We compared visual outcomes in these two groups and rates of subsequent postoperative complications. Baseline characteristics of the groups were similar. Compared with the control group, the PSB group had statistically significant lower rates of proliferative vitreoretinopathy (PVR) (5.3% versus 38.4%, P < 0.05) and a trend towards lower rates of RD (15.8% versus 41.1%, P = 0.1). PSB group eyes had a statistically significant improvement of their best distance visual acuity (BDVA) with lower means of final BDVA-grade (P < 0.05) and logMAR vision (P < 0.05). Eyes in the control group had no improvement in these parameters. PSB procedure during posterior segment OGI repair may decrease the risk of subsequent retinal complications and improve final visual outcome.


[Research plan to study allelopathy under microgravity].
Living organisms interact each others and form ecological system on the earth. Such interactions between organisms and species have been long known, and studied at a global scale. Allelopathy is a phenomenon observed in many plants that emit specific chemicals acting on other organisms, including animals and microorganisms, in either inhibitory or excitatory ways. We propose to study whether phenomena of allelopathy are modified under altered gravity or not. If biosynthesis, emission and sensing mechanism of allelopathic substances would be affected by gravity, many organisms and ecological system might show different behaviors based on the inter-organisms and species interactions under microgravity. In the macroscopic scale, transport of the substances between organisms is largely affected by convection induced by gravity. Furthermore, the fate of allelopathic substances in confined environment differs from that seen on the earth, because of lacking sink compartment for removal and producing exotic bio-active substances by man-made system. We design basic ground experiment to evaluate gravitational effects on allelopathy applying pseudo-microgravity. Our study contributes to the synthesis of ecological system and its control on spacecrafts and extraterrestrial bodies. It also makes possible to sustain qualitative human life even on the ground under confined artificial environment that dominates in many scenes.


Confidence intervals for intraclass correlation coefficients in a nonlinear dose-response meta-analysis.
This work is motivated by a meta-analysis case study on antipsychotic medications. The Michaelis-Menten curve is employed to model the nonlinear relationship between the dose and D2 receptor occupancy across multiple studies. An intraclass correlation coefficient (ICC) is used to quantify the heterogeneity across studies. To interpret the size of heterogeneity, an accurate estimate of ICC and its confidence interval is required. The goal is to apply a recently proposed generic beta-approach for construction the confidence intervals on ICCs for linear mixed effects models to nonlinear mixed effects models using four estimation methods. These estimation methods are the maximum likelihood, second-order generalized estimating equations and two two-step procedures. The beta-approach is compared with a large sample normal approximation (delta method) and bootstrapping. The confidence intervals based on the delta method and the nonparametric percentile bootstrap with various resampling strategies failed in our settings. The beta-approach demonstrates good coverages with both two-step estimation methods and consequently, it is recommended for the computation of confidence interval for ICCs in nonlinear mixed effects models for small studies.


Stakeholder opinions on value in healthcare.
To determine how the value of new medicines is defined from the perspective of different stakeholders in nine European countries. We carried out an explorative survey by conducting in-depth qualitative interviews with a range of experts with health technology assessment, clinical provision, health economics, payer, academic and industry experience. Experts were asked to distribute 100 points over a predefined list of attributes related to value. In total, 30 interviews were conducted with seven academics, five clinicians, nine economists, five payers and four providers. Each of these categories was represented in responses from France, Germany, the Netherlands and the UK. Comments on the interview guide were all positive. There was substantial variation in definitions of value provided by respondents. Most respondents indicated that delivering benefits to the patient is a prerequisite for a healthcare product to be of value, but only half included costs or resource use in their definition. Quantitative responses showed that effectiveness and efficacy were considered the most important attributes of value, followed by safety and side effects. Lower scores were given to other attributes, such as dignity and individualism, and invasiveness of the treatment. Resources and costs associated with complications and readmissions were considered important by academics and economists, but not clinicians. The interview findings indicate that the value of a drug is subjective in being perceived differently by different stakeholders, and for different treatment types. Future research will require evaluation of the views of a larger number of stakeholders in more and different countries to confirm these findings.


Role of passenger leukocytes in allograft rejection: effect of depletion of donor alveolar macrophages on the local production of TNF-alpha, T helper 1/T helper 2 cytokines, IgG subclasses, and pathology in a rat model of lung transplantation.
Acute lung allograft rejection is believed to be initiated by passenger leukocytes, such as alveolar macrophages (AM), in the donor organ, which release TNF-alpha, and present alloantigens to host lymphocytes, to up-regulated Th1 cellular and humoral immunity. However, the role of donor AM in local TNF-alpha synthesis, and their ability to induce local Th1 cellular and humoral immunity have not been evaluated. By depleting Brown Norway (BN, RT1n) rat lung allografts of AM before transplantation into Lewis rat (LEW, RT1(1)) recipients, the current study determined the role of donor AM in including the production of TNF-alpha, IFN-gamma (Th1 cytokine), IL-4 (Th2 cytokine), IgG subtypes, and rejection pathology in the allograft. The data show that compared with untreated BN allografts, pretransplant depletion of donor lung AM resulted in significantly less TNF-alpha, and IFN-gamma production in allograft bronchoalveolar lavage fluid with variable effects on local IL-4 production. Depletion of AM in the donor lung before transplantation affected the local production of several IgG subclasses. However, pretransplant depletion of donor AM had no effect on the development of the pathology of severe acute rejection. These data show that donor AM have a central role in the local synthesis of TNF-alpha and induce the production of IFN-gamma and IgG subtypes, locally, during acute lung allograft rejection. However, depletion of AM before transplantation does not prevent the development of severe acute rejection in BN rat lungs, transplanted into LEW recipients.


Emergence Patterns of Rare Arable Plants and Conservation Implications.
Knowledge on the emergence patterns of rare arable plants (RAP) is essential to design their conservation in Europe. This study hypothesizes that is possible to find functional groups with similar emergence patterns within RAP with the aim of establishing management strategies. Seeds of 30 different species were collected from Spanish arable fields and sown under two tillage treatments: (a) 1 cm depth without soil disturbance to simulate no-till, and (b) 1-10 cm depth with soil disturbance every autumn to simulate tillage to 10 cm depth. Two trials were established; the first trial being maintained for three seasons and the second for two seasons. Relative emergence in autumn, winter and spring was calculated each season. Afterwards, multivariate analysis was performed by K-means clustering and Principal Component Analysis to find groups of RAP species with similar emergence patterns. Four RAP groups were defined, and each was based on its main emergence season: autumn, winter, spring, or autumn-winter. Tillage treatment and the year of sowing had little effect on emergence patterns, which were mostly dependent on environmental factors, particularly temperature and rainfall. Therefore, conservation strategies could be designed for each of these RAP functional groups based on emergence patterns, rather than on a species-by-species basis.


Integrins in regulation of tissue development and function.
Cell adhesion is indispensable for embryonic development and for proper tissue function. In metazoans, integrins are the major adhesion receptors that connect cells to components of the extracellular matrix. Integrins are implicated in assembly of extracellular matrices, cell adhesion and migration on extracellular matrices, and in vertebrates (in which the integrin family has expanded) they can also mediate cell-cell adhesion. Furthermore, integrin-mediated adhesion can modulate many different signal transduction cascades and support cell survival, proliferation, and influence the expression of differentiation-related genes. In this review we briefly explain how integrins can affect so many different aspects of cell behavior and discuss evidence for roles of integrins in tissue development, function, and disease.


Examining Physician Interactions with Disease Advocacy Organizations.
Disease advocacy organizations (DAOs) have traditionally focused on raising awareness of rare diseases, providing educational resources to patients, and supporting patients and families. Previous research has described how scientists collaborate with DAOs, but few empirical data are available regarding the extent to which physicians interact with DAOs and how those interactions impact patient care. We conducted a national survey of 230 board-certified pediatric neurologists to assess their engagement with DAOs and their beliefs about the impact of DAOs on patient care. In that context, we evaluated a set of 24 items describing interactions between physicians and DAOs. Exploratory factor analysis produced a 19-item model capturing four types of physician-DAO engagement: (1) accessing or distributing DAO-produced materials (6 items, alpha = 0.80); (2) consulting on DAO activities (5 items, alpha = 0.81); (3) collaborating with DAOs on research activities (6 items, alpha = 0.80); and (4) co-producing scholarly materials with DAOs (2 items, alpha = 0.80). Our data indicate that physicians engage with DAOs in more frequent and diverse ways than has been previously reported. Almost all physicians in our sample had interacted directly with a DAO in some way, from low-effort activities such as visiting a DAO's website to deeper forms of engagement including coauthoring journal articles. These findings may provide a framework for bioethicists to characterize the nature and extent of physician interactions with advocacy organizations, which is critical for evaluating the ethical implications of physician-DAO relationships.


How Close Are We to Profiling Immunogenicity Risk Using In Silico Algorithms and In Vitro Methods?: an Industry Perspective.
In silico HLA-binding algorithms and in vitro T cell-based assays as predictive tools for human immunogenicity risk have made inroads in the biotherapeutic drug discovery and development process. Currently, these tools are being used only for candidate selection or characterization and not for making a go/no-go decision for further development. A clear limitation for a broader implementation is the lack of correlation between the predicted T cell epitope content/immune reactivity potential of a biotherapeutic and the subsequent ADA-related clinical immunogenicity outcome. The current state of technologies and their pros and cons were discussed as a part of the 2016 AAPS National Biotechnology Conference in a themed session. A review of the advances in the area and the session talks along with the ensuing discussions are summarized in this commentary.


Survey of comprehensive accident and injury experience of high school students in Saskatchewan.
Accidents and injuries typically have been studied on the basis of hospital and coroners' reports, by age group, activity, and accident location. The comprehensive accident experience of population groups has not been reported, although Guyer and Gallagher have attempted an estimate for childhood injuries. The accident and injury experience during one year (1982) of all 746 grade 11 and 12 students in three Saskatchewan high schools was surveyed by the Saskatchewan Child Safety Committee. In Part A of the survey, comprehensive information was gathered on the extent and types of accidents and injuries and modes of medical treatment. In Part B of the survey, employment injuries from the summer of 1982 were reviewed. When Part B injuries were used to represent occupational injuries in Part A of the survey, work became the primary source of injuries to students during the year.


Effects of a preferential myosin loss on Ca2+ activation of force generation in single human skeletal muscle fibres.
Preferential loss of the motor protein myosin, as observed in patients with acute quadriplegic myopathy (AQM) or cancer cachexia, causes generalized muscle wasting, muscle weakness and a decrease in muscle fibre force normalized to cross-sectional area. It remains unclear, however, whether this myosin loss influences other important features of muscle fibre function, such as Ca(2+) activation of the contractile proteins. To address this question, we have studied Ca(2+) sensitivity of force generation using skinned muscle fibres from four patients with AQM or cancer cachexia and a preferential loss of myosin; and from seven healthy control individuals. Force and apparent rate constant of force redevelopment (k(tr)) were assessed in solutions with varying Ca(2+) concentrations (pCa), allowing construction of relative force-pCa and k(tr)-pCa relationships. Results showed a rightward shift of the relative force-pCa relationship and a leftward shift of the relative k(tr)-pCa curve in muscle fibres with a preferential myosin loss. To improve the understanding of the mechanisms underlying these alterations, the relative stiffness-pCa relationship was evaluated. A rightward shift of this curve was observed, suggesting that the changes in the Ca(2+) activation of force and k(tr) were predominantly due to a decrease in the relative number of attached cross-bridges at different pCa values. Thus, a change in Ca(2+) activation of the contractile apparatus in patients with preferential myosin loss is proposed as an additional factor contributing to the muscle function impairment in these patients.


Anaphylaxis and sport.
This article aims to review the most relevant studies on exercise-induced anaphylaxis, published in the last year, in order to provide comprehensive and updated evidence and hopefully contribute to a better definition of its pathophysiological mechanisms and treatment strategies. The search strategy was performed from 1/2/2013 to 31/1/2014 by scanning the principal electronic bibliographic database and by hand-searching the main scientific publications in Allergy and Clinical Immunology. Among the identified articles, 17 articles were selected to be part of the systematic review. Eligible studies included five experimental trials, eight case reports and four letters. The overall collected evidence was of very low quality. No randomized controlled trials were identified by the searching process. Most of the data derived from reports performed in small population samples or even in individual cases. Except for one article addressing issues related to the preventive management of exercise-induced anaphylaxis, all other articles focused on prevalence rates, causative triggers and pathogenetic mechanisms. More interesting findings were related to the influence of the IL-4-C590T polymorphism on the onset of wheat-dependent exercise-induced anaphylaxis and to the usefulness of the immuno solid-phase allergen chip technique in the allergic screening of polysensityzed athletes at risk of severe reactions.


Mitochondrial dysfunction in age-related macular degeneration: melatonin as a potential treatment.
Introduction: Age-related Macular Degeneration (AMD), a retinal neurodegenerative disease is the most common cause of blindness among the elderly in developed countries. The impairment of mitochondrial biogenesis has been reported in human retinal pigment epithelium (RPE) cells affected by AMD. Oxidative/nitrosative stress plays an important role in AMD development. The mitochondrial respiratory system is considered a major site of reactive oxygen species (ROS) generation. During aging, insufficient free radical scavenger systems, impairment of DNA repair mechanisms and reduction of mitochondrial degradation and turnover contribute to the massive accumulation of ROS disrupting mitochondrial function. Impaired mitochondrial function leads to the decline in the autophagic capacity and induction of inflammation and apoptosis in human RPE cells affected by AMD.Areas covered: This article evaluates the ameliorative effect of melatonin on AMD and examines AMD pathogenesis with an emphasis on mitochondrial dysfunction. It also considers the potential effects of melatonin on mitochondrial function.Expert opinion: The effect of melatonin on mitochondrial function results in the reduction of oxidative stress, inflammation and apoptosis in the retina; these findings demonstrate that melatonin has the potential to prevent and treat AMD.


The Complex Genetic Basis of Congenital Heart Defects.
Twenty years ago, chromosomal abnormalities were the only identifiable genetic causes of a small fraction of congenital heart defects (CHD). Today, a de novo or inherited genetic abnormality can be identified as pathogenic in one-third of cases. We refer to them here as monogenic causes, insofar as the genetic abnormality has a readily detectable, large effect. What explains the other two-thirds? This review considers a complex genetic basis. That is, a combination of genetic mutations or variants that individually may have little or no detectable effect contribute to the pathogenesis of a heart defect. Genes in the embryo that act directly in cardiac developmental pathways have received the most attention, but genes in the mother that establish the gestational milieu via pathways related to metabolism and aging also have an effect. A growing body of evidence highlights the pathogenic significance of genetic interactions in the embryo and maternal effects that have a genetic basis. The investigation of CHD as guided by a complex genetic model could help estimate risk more precisely and logically lead to a means of prevention.


Implantable biomaterial based on click chemistry for targeting small molecules.
Specific and targeted delivery of medical therapies continues to be a challenge for the optimal treatment of multiple medical conditions. Technological advances permit physicians to target most sites of the body. However, after the intervention, physicians rely on systemic medications that need frequent dosing and may have noxious side effects. A novel system combining the temporal flexibility of systemic drug delivery and the spatial control of injectable biomaterials would improve the spatiotemporal control of medical therapies. Here we present an implantable biomaterial that harnesses in vivo click chemistry to enhance the delivery of suitable small molecules by an order of magnitude. The results demonstrate a simple and modular method to modify a biomaterial with small molecules in vitro and present an example of a polysaccharide modified hours after in vivo implantation. This approach provides the ability to precisely control the moment when biochemical and/or physical signals may appear in an implanted biomaterial. This is the first step towards the construction of a biomaterial that enhances the spatial location of systemic small molecules via in vivo chemical delivery.


The role of hydrogen peroxide-inducible clone-5 in tumor progression.
The poor prognosis of cancers such as hepatocellular carcinoma is due to high recurrence rate mainly caused by metastasis. Target therapy aiming at critical signal molecules within these pathways is one of the promising strategies for the prevention of metastasis. Hydrogen peroxide-inducible clone-5 (Hic-5), which belongs to the paxillin superfamily, is emerging as a potential target along the metastatic signaling pathway. Hic-5 and paxillin share similar structural features; however, there are a lot of different biochemical properties between them, including tissue-specific distribution, regulation of gene expression, critical signal cascade, and the impacts on cellular phenotypes. This review focus on the recent studies of Hic-5 related to its impacts on signal transduction and transcription responsible for tumor progression. Hic-5 may regulate mitogen-activated protein kinase cascade for cell migration and invasion in various systems. Hic-5 can mediate transforming growth factor-β1-induced epithelial-mesenchymal transition (EMT) via RhoA- and Src-dependent signaling. Moreover, Hic-5 plays a central role in a positive feedback Hic-5-NADPH oxidase-ROS-JNK signal cascade. This sustained signaling is required for regulating EMT-related genes including E-cadherin, Snail, MMP9, and Zeb-1. In addition, Hic-5 can be a transcription coregulatory factor for a lot of nuclear receptors. Owing to the critical role of Hic-5 in signal transduction and transcription responsible for tumor progression, it can be a potential therapeutic target for the prevention of tumor metastasis.


Evaluation of hospital inpatient complications: a planning approach.
Hospital inpatient complications are one of a number of adverse health care outcomes. Reducing complications has been identified as an approach to improving care and saving resources as part of the health care reform efforts in the United States.An objective of this study was to describe the Potentially Preventable Complications software developed as a tool for evaluating hospital inpatient outcomes. Additional objectives included demonstration of the use of this software to evaluate the connection between health care outcomes and expenses in United States administrative data at the state and local levels and the use of the software to plan and implement interventions to reduce hospital complications in one U.S. metropolitan area. The study described the Potentially Preventable Complications software as a tool for evaluating these inpatient hospital outcomes. Through administrative hospital charge data from California and Maryland and through cost data from three hospitals in Syracuse, New York, expenses for patients with and without complications were compared. These comparisons were based on patients in the same All Patients Refined Diagnosis Related Groups and severity of illness categories. This analysis included tests of statistical significance.In addition, the study included a planning process for use of the Potentially Preventable Complications software in three Syracuse hospitals to plan and implement reductions in hospital inpatient complications. The use of the PPC software in cost comparisons and reduction of complications included tests of statistical significance. The study demonstrated that Potentially Preventable Complications were associated with significantly increased cost in administrative data from the United States in California and Maryland and in actual cost data from the hospitals of Syracuse, New York. The implementation of interventions in the Syracuse hospitals was associated with the reduction of complications for urinary tract infection, decubitus ulcer, and pulmonary embolism. The study demonstrated that the Potentially Preventable Complications software could be used to evaluate hospital outcomes and related costs at the aggregate and diagnosis specific levels. It also indicated that the system could be used to plan and implement interventions to improve outcomes on an individual or multihospital basis.


Size of CA1-evoked synaptic potentials is related to theta rhythm phase in rat hippocampus.
Cholinergic and GABAergic neurons projecting to the hippocampus fire with specific phase relations to theta rhythm oscillations in the electroencephalogram (EEG). To determine if this phasic input has an impact on synaptic transmission within the hippocampus, we recorded evoked population excitatory postsynaptic potential (EPSPs) during different phases of theta rhythm by using techniques similar to those described in Rudell and Fox. Synaptic potentials elicited by stimulation of region CA3 of the contralateral hippocampus were recorded in region CA1 and CA3. In these experiments, the initial slope of evoked potentials showed a change in magnitude during different phases of the theta rhythm recorded in the dentate fissure, with individual trials showing an average of 9.5% change in slope of potentials, and the average across all experiments showing a change of 7.8%. Evoked potentials were maximal 18 degrees after the positive peak of the dentate fissure theta EEG. These potentials were also smaller by 18.2% during theta as opposed to non-theta states. Phasic changes in modulation of synaptic transmission could contribute to phase precession of hippocampal place cells and could enhance storage of new sequences of activity as demonstrated by computational models.


Probing Peptidylprolyl Bond cis/trans Status Using Distal 19 F NMR Reporters.
A method for measuring peptidylprolyl bond cis-trans conformational status in peptide models is described, using 4-fluorophenylalanine (4FPhe) as a distal reporter for 19 F NMR. The %cis-Pro population was measured for peptides of the general structure Ac-X-Pro-Z-Ala-Ala-4FPhe (X and Z are proteinogenic amino acids) at pH 7.4, and provided conformational populations consistent with literature values obtained by more complex methods. This approach was applied to probe the prolyl bond status in pentapeptide models of the intrinsically disordered C-terminal region of α-synuclein, which mirrored the preferences in the Ac-X-Pro-Z-Ala-4FPhe models. Advantageously, the 19 F reporter group does not need to be adjacent to or attached to proline to provide quantifiable signals and distal 4-fluorophenylalanines can be placed so as not to influence prolyl bond conformation. Finally, we demonstrated that the prolyl bond status is not significantly affected by pH when there are ionisable amino acid residues at the carboxyl side of proline, which makes 19 F NMR an invaluable tool with which to study proline isomerism at a range of pHs and in different solvents and buffers.


Replacing a lectin domain residue in L-selectin enhances binding to P-selectin glycoprotein ligand-1 but not to 6-sulfo-sialyl Lewis x.
Selectin-ligand interactions (bonds) mediate leukocyte rolling on vascular surfaces. The molecular basis for differential ligand recognition by selectins is poorly understood. Here, we show that substituting one residue (A108H) in the lectin domain of L-selectin increased its force-free affinity for a glycosulfopeptide binding site (2-GSP-6) on P-selectin glycoprotein ligand-1 (PSGL-1) but not for a sulfated-glycan binding site (6-sulfo-sialyl Lewis x) on peripheral node addressin. The increased affinity of L-selectinA108H for 2-GSP-6 was due to a faster on-rate and to a slower off-rate that increased bond lifetimes in the absence of force. Rather than first prolonging (catching) and then shortening (slipping) bond lifetimes, increasing force monotonically shortened lifetimes of L-selectinA108H bonds with 2-GSP-6. When compared with microspheres bearing L-selectin, L-selectinA108H microspheres rolled more slowly and regularly on 2-GSP-6 at low flow rates. A reciprocal substitution in P-selectin (H108A) caused faster microsphere rolling on 2-GSP-6. These results distinguish molecular mechanisms for L-selectin to bind to PSGL-1 and peripheral node addressin and explain in part the shorter lifetimes of PSGL-1 bonds with L-selectin than P-selectin.


Exploring the molecular mechanisms of OSU-03012 on vascular smooth muscle cell proliferation.
Restenosis is resulted from the proliferation and migration of vascular smooth muscle cells (VSMCs) from the arterial media into the intima within the vessel lumen following percutaneous transluminal coronary angioplasty (PTCA). OSU-03012, a synthetic compound (2-amino-N-{4-[5-(2-phenanthrenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]-phenyl} acetamide) acting as a PDK-1 inhibitor, is used as an apoptosis-promoting anticancer drug. However, whether OSU-03012 can inhibit VSMC proliferation and migration following PTCA remains unclear. In this study, we used A10 smooth muscle cells cultured in 10% FBS for stimulating proliferation and evaluated the inhibitory effects of OSU-03012 on cell proliferation and migration. The data demonstrated that OSU-03012 dose-dependently inhibited A10 cell proliferation examined by Trypan blue, MTT and morphological alteration assays, and inhibited the levels of proliferation-related proteins, proliferating cell nuclear antigen (PCNA), phosphorylated ERK examined by western blotting. Additionally, 10 μM OSU-03012 also enhanced apoptosis examined using DAPI assay by regulating apoptosis-related proteins. Furthermore, compared with the control group, A10 cells treated with 10 μM OSU-03012 showed a lower number of migrating cells examined by Boyden Chamber assay, and a dose-dependently reduced NFκB-dependent and interferon-stimulated response element (ISRE) promoter luciferase activities, implying the anti-migration and anti-inflammation effects of OSU03012. Taken together, this study provides insights into the pharmacological mechanisms of OSU-03012 in preventing smooth muscle cell proliferation, migration, and inflammation supporting the novel discovery of OSU-03012 as an adjuvant therapy for balloon injury-induced restenosis.


Uterine activity in induced labour.
A study was made of the characteristics of oxytocin-induced labour. Twelve patients with, and eight without membrane rupture were closely matched for factors likely to influence the character and duration of labour. In both groups uterine activity increased until a stable state was achieved loowing which there was little alteration until delviery. The duration of labour was shorten in the group with ruptured membranes as compared with intact membranes although, paradoxically, the uterineactivity tended to be greater when the membranes were intact.


A time course analysis of the electrophysiological properties of neurons differentiated from human induced pluripotent stem cells (iPSCs).
Many protocols have been designed to differentiate human embryonic stem cells (ESCs) and human induced pluripotent stem cells (iPSCs) into neurons. Despite the relevance of electrophysiological properties for proper neuronal function, little is known about the evolution over time of important neuronal electrophysiological parameters in iPSC-derived neurons. Yet, understanding the development of basic electrophysiological characteristics of iPSC-derived neurons is critical for evaluating their usefulness in basic and translational research. Therefore, we analyzed the basic electrophysiological parameters of forebrain neurons differentiated from human iPSCs, from day 31 to day 55 after the initiation of neuronal differentiation. We assayed the developmental progression of various properties, including resting membrane potential, action potential, sodium and potassium channel currents, somatic calcium transients and synaptic activity. During the maturation of iPSC-derived neurons, the resting membrane potential became more negative, the expression of voltage-gated sodium channels increased, the membrane became capable of generating action potentials following adequate depolarization and, at day 48-55, 50% of the cells were capable of firing action potentials in response to a prolonged depolarizing current step, of which 30% produced multiple action potentials. The percentage of cells exhibiting miniature excitatory post-synaptic currents increased over time with a significant increase in their frequency and amplitude. These changes were associated with an increase of Ca2+ transient frequency. Co-culturing iPSC-derived neurons with mouse glial cells enhanced the development of electrophysiological parameters as compared to pure iPSC-derived neuronal cultures. This study demonstrates the importance of properly evaluating the electrophysiological status of the newly generated neurons when using stem cell technology, as electrophysiological properties of iPSC-derived neurons mature over time.


The identification of functional modules from the genomic association of genes.
By combining the pairwise interactions between proteins, as predicted by the conserved co-occurrence of their genes in operons, we obtain protein interaction networks. Here we study the properties of such networks to identify functional modules: sets of proteins that together are involved in a biological process. The complete network contains 3,033 orthologous groups of proteins in 38 genomes. It consists of one giant component, containing 1,611 orthologous groups, and of 516 small disjointed clusters that, on average, contain only 2.7 orthologous groups. These small clusters have a homogeneous functional composition and thus represent functional modules in themselves. Analysis of the giant component reveals that it is a scale-free, small-world network with a high degree of local clustering (C = 0.6). It consists of locally highly connected subclusters that are connected to each other by linker proteins. The linker proteins tend to have multiple functions, or are involved in multiple processes and have an above average probability of being essential. By splitting up the giant component at these linker proteins, we identify 265 subclusters that tend to have a homogeneous functional composition. The rare functional inhomogeneities in our subclusters reflect the mixing of different types of (molecular) functions in a single cellular process, exemplified by subclusters containing both metabolic enzymes as well as the transcription factors that regulate them. Comparative genome analysis, thus, allows identification of a level of functional interaction between that of pairwise interactions, and of the complete genome.


Some Preformulation Studies of Pyruvic Acid and Other α-Keto Carboxylic Acids in Aqueous Solution: Pharmaceutical Formulation Implications for These Peroxide Scavengers.
The purpose of this study is to assess some of the variables determining the aldol-like condensation of pyruvic acid (1), a peroxide scavenger, in aqueous solution to parapyruvic acid and higher oligomers. Its stability is compared to 3 other α-keto carboxylic acids, 2 with sterically hindered methylene groups alpha to the keto functionality (2-3) and phenylglyoxylic acid (4) with no methylene group. High-performance liquid chromatography, nuclear magnetic resonance, and liquid chromatography mass spectroscopy techniques are used in the kinetics and product analyses. 1 condensation is concentration dependent and base catalyzed above pH 7, consistent with the reaction mechanism proceeding through the attack of the fraction of the methylene group, alpha to the keto group, in its anionic form, at the keto group of a second molecule of 1. The major product is confirmed to be parapyruvic acid, but higher-order oligomers are also observed. All 3 of the other α-keto carboxylic acids 2-4 are considerably less reactive, with 4 being completely stable. Stable solutions of 1 can be prepared by the use of relatively dilute solutions maintained at slightly acidic pH values. 1 prevents the oxidation of methionine on addition of hydrogen peroxide.


[Colonic intussusception by leiomyosarcoma].
We report the case of female patient, 34 years old, occupation Secretary. Polycystic ovary and chronic anemia. No family history of cancer. The patient reported three weeks abdominal pain, fever, bloody loose stools, nausea and vomiting. Abdominal pain is located in flank and right lower quadrant, is colicky, intensity 5 / 10, associated with abdominal distension. On physical examination, vital functions indicated T 38.6 ° FC 98 x min, FR 18 x min, BP 120/80, was awake, she looked pale, in generally fair condition, lucid and oriented. The abdomen was distended, the increased bowel sounds, tympanic to percussion, with tenderness in the right abdomen (more intense in the right iliac fossa), palpable mass is not defined edges lower right quadrant of about 6cm. abdominal pain syndrome (appendicular mass, intestinal obstruction, intussusceptions). In examinations auxiliars highlighted in 9.1 g of hemoglobin with decreased corpuscular constants. Reviewed by history "rectal bleeding", DRE: yellow stool, no trace of blood, so that colonoscopy was deferred. Reassessed at 24 hours, we decide surgery with a presumptive diagnosis of intestinal obstruction, intussusception. In surgery, we identified a transverse colon tumor (with colo-colonic intussusception) and 10cm of colon was resected and meso tumor: TT anastomosis was performed in transverse colon. The lesion was a tumor of the middle region of the transverse colon, proliferative fibroid appearance, which almost completely obstructed the intestinal lumen, measuring about 7x5cm. The study of pathology with immunohistochemistry indicated that the tumor corresponded to leiomyosarcoma of the colon. Was discharged in good condition. The rarity of this type of malignancy and this type of presentation led us to make this report.


Altered outward-rectifying K(+) current reveals microglial activation induced by HIV-1 Tat protein.
Microglial cells are believed to be one of the key elements in the development of the HIV-related neuropathology. Not only can microglial cells be productively infected by the virus, but they are also sensitive to viral proteins. Among them, the HIV-1 regulatory protein Tat, which was shown to have neurotoxic activity, is able to promote some proinflammatory functions of microglia. Considering that microglial activation goes along with a change of ion channel profile, we aimed to study whether Tat could influence microglial electrophysiology. When microglial cultures obtained from neonatal rats were treated with Tat (> or = 100 ng/ml), whole-cell recording showed the appearance of a large outwardly rectifying current (OR) virtually absent in untreated control cells. According to voltage dependence of the kinetic variables, K(+) permeability, and pharmacological sensitivity, the Tat-induced current was due to the presence of functional Kv1.3 channels. The effect of Tat was abolished by specific anti-Tat polyclonal antibody and by heat denaturation of Tat protein, confirming that the OR enhancement was due to the viral protein. Interestingly, the OR current induced by Tat was largely prevented by two inhibitors of the transcription factor NF-kappaB, TPCK and SN50, which suggests an involvement of NF-kappaB in the effect of the viral protein. The relatively high dose of Tat needed to observe an effect (> or = 100 ng/ml) might indicate that the action of Tat required entrance of the protein into the cell, rather than being mediated by a membrane receptor. In conclusion, the HIV-1 protein Tat is able to enhance OR K(+) current in rat microglia through a mechanism involving the activation of NF-kappaB. We propose that such effect of Tat could be part of the process of microglial activation known to take place in the brain of persons with neuro-AIDS.


Chronopharmacology of oxacalcitriol in rat model of osteoporosis.
We have previously reported the merits of chronopharmacological effect of 1-alpha(OH) vitamin D3 in aged stroke-prone spontaneously hypertensive rat (SHRSP), a model of osteoporosis [Eur. J. Pharmacol. 428 (2001) 283.]. In this study, the chronopharmacological effect of 22-oxacalcitriol, a newly developed active vitamin D3 analogue with less calcemic activity, was evaluated by a single and repeated dosing of the drug in aged SHRSP. Animals (7 months old) were kept in rooms with a 12-h light/dark cycle. Single (12.5 microg/kg, i.v.) and repeated (5 microg/kg, i.v. three times a week for 12 weeks) dosing of 22-oxacalcitriol or vehicle was given at either 2 h after lights on (2HALO) or 14 h after lights on (14HALO). The severity of adverse reactions such as the changes of body weight, hypercalcemia and hyperphosphatemia, was significantly mild when the drug was given at 14HALO. Especially, the increase of serum Ca concentration was not detected at 14HALO trial. Serum concentrations of total (protein-bound and unbound) 22-oxacalcitriol and albumin (a major binding protein of the drug) of the 2HALO and 14HALO trials did not significantly differ. The decrease of parathyroid hormone (PTH) concentration was greater in the 14HALO trial while the increase in urinary ratio of Ca to creatinine was greater in the 2HALO trial. The increase in bone density of both femurs at the end of the study was greater in the 14HALO trial. The suppression of urinary excretion of deoxypyridinoline, an index of bone resorption capacity of osteoclast, was greater in the 14HALO trial, which indicates that the efficacy of 22-oxacalcitriol for suppressing bone resorption might vary with the dosing time. This is the first study to show the dosing-time-dependent changes in the efficacy and toxicity of 22-oxacalcitriol in the animal model of osteoporosis. Chronopharmacological differences seem to be more prominent than those of other vitamin D analogues. To use 22-oxacalcitriol at an adequate timing might provide better efficacy and safety than other vitamin D3 analogues for the treatment of osteoporosis.


[Limiting the growth of Saccharomyces serevisiae yeasts under chemostat conditions by carbon and nitrogen sources].
Chemostat Saccharomyces cerevisiae cultures were obtained at D = 0.066 h-1 and growth limitation with ethanol and a nitrogen source with similar population densities. The threshold concentration of ethanol was 0.1 g/L and that of nitrogen 0.014 g/L. The optimal specific load of ethanol was 2.5--3.5 g/g biomass per hour with the economic coefficient of 40%. At the constant specific ethanol load, the economic coefficient changed at different rates of flow. The cultures differed in the content and amino acid composition of protein.


Initial Characterization of the Growth Stimulation and Heat-Shock-Induced Adaptive Response in Developing Lake Whitefish Embryos after Ionizing Radiation Exposure.
Ionizing radiation is known to effect development during early life stages. Lake whitefish (Coregonus clupeaformis) represent a unique model organism for examining such effects. The purpose of this study was to examine how ionizing radiation affects development in lake whitefish embryos and to investigate the presence of an adaptive response induced by heat shock. Acute exposure to 137Cs gamma rays was administered at five time points corresponding to major developmental stages, with doses ranging from 0.008 to 15.5 Gy. Chronic gamma-ray exposures were delivered throughout embryogenesis within a custom-built irradiator at dose rates between 0.06 and 4.4 mGy/day. Additionally, embryos were given a heat shock of 3, 6 or 9°C prior to a single acute exposure. Radiation effects were assessed based on survival, development rate, morphometric measurements and growth efficiency. Embryos showed high resistance to acute exposures with an LD50/hatch of 5.0 ± 0.7 Gy immediately after fertilization, increasing to 14.2 ± 0.1 Gy later in development. Chronic irradiation at all dose rates stimulated growth, with treated embryos up to 60% larger in body mass during development compared to unirradiated controls. Chronic irradiation also accelerated the time-to-hatch. A heat shock administered 6 h prior to irradiation reduced mortality by up to 25%. Overall, low-dose chronic irradiation caused growth stimulation in developing lake whitefish embryos and acute radiation mortality was reduced by a heat-shock-induced adaptive response.


Combined nitrous oxide 70% with intranasal fentanyl for procedural analgosedation in children: a prospective, randomised, double-blind, placebo-controlled trial.
Nitrous oxide 70% (N2O 70%) is an excellent medication for procedural analgosedation (PAS), yet the limit of its analgesic power remains uncertain; therefore, a combination with intranasal fentanyl (INF) was suggested. However, this combination seems to result in a higher rate of vomiting and deeper sedation. This study aimed at assessing the analgesic efficacy, sedation depth and rate of adverse events of PAS with N2O 70% with and without INF. Patients aged 2-16 years who qualified for PAS with N2O 70% were randomly assigned to receive either INF or placebo prior to N2O inhalation in this randomised, double-blind study, which was performed in a tertiary children's hospital ED between September 2015 and October 2017. Behaviour during the procedure was evaluated using the Face, Leg, Activity, Cry and Consolability (FLACC) scale and the Modified Behavioural Pain Scale (MBPS); analgesic efficacy was assessed with a self-reported pain scale. Sedation depth using the validated University of Michigan Sedation Scale and adverse events in the ED and during the following 12 hours were documented. A total of 402 patients were included; 3 did not tolerate N2O and therefore had to be excluded. Overall, 399 patients were analysed, of whom 201 (50.4%) received INF. No significant group differences with regard to FLACC scale score, self-reported pain, MBPS score and sedation depth were found. In addition, the two groups did not differ with regard to all types of adverse events. Combining N2O 70% with INF resulted in no differences with regard to FLACC scale score, self-reported pain, MBPS score, patient and parental satisfaction rate, sedation depth, and adverse events. NCT02533908.


Ocular side effects from herbal medicines and nutritional supplements.
To review the more significant herbal and nutritional agents of clinical importance to ophthalmologists and describe the ocular side effects for each. World Health Organization (WHO) classification and guidelines for clinicians are provided. Retrospective observational case series. A retrospective observational case series of reports of ocular side effects or systemic side effects from medications used for the eye from herbal medicines and nutritional supplements. Cases were collected from spontaneous reports submitted to the WHO, the Food and Drug Administration, and the National Registry of Drug-Induced Ocular Side Effects. A review of the world's literature was performed to obtain additional case reports and insight into adverse ocular reactions. Data were collected on age, gender, duration of therapy, concomitant medications, dosage, and dechallenge and rechallenge results. The National Registry of Drug-Induced Ocular Side Effects received 263 spontaneous reports, in addition to 60 case reports from the literature. Canthaxanthine, chamomile, Datura, Echinacea purpurea, Ginkgo biloba, licorice, niacin, and vitamin A are all associated with clinically significant ocular side effects. Herbal medicines and nutritional supplements can cause ocular side effects. Clinicians need to recognize these adverse events, because a large segment of the population uses them, many times without the treating physician's knowledge.


Sarcopenic obesity, weight loss, and mortality: the English Longitudinal Study of Ageing.
Background: Age-related sarcopenia describes the loss of muscle strength and often accompanies an increase in adiposity in the elderly.Objective: We examined the association of sarcopenic obesity and changes in muscle strength and weight with the risk of mortality.Design: Participants (n = 6864) were community-dwelling men and women (45.6% men; 54.4% women; mean ± SD age: 66.2 ± 9.5 y) from the English Longitudinal Study of Ageing. Handgrip strength and body mass index (BMI; in kg/m2) were measured at baseline and at a 4-y follow-up. Individual participant data were linked with death records from National Health Service registries. Sarcopenic obesity was defined as obese individuals (BMI ≥30) in the lowest tertile of sex-specific grip strength (<35.3 kg for men and <19.6 kg for women).Results: There were 906 deaths over a mean follow-up of 8 y. Compared with the reference group (normal BMI and highest handgrip tertile), the risk of all-cause mortality increased as grip strength reduced within each BMI category. For participants in the lowest handgrip tertile, there was little difference in the risk between normal BMI (HR: 3.25; 95% CI: 1.86, 5.65), overweight (HR: 2.50; 95% CI: 1.44, 4.35), and obesity (HR: 2.66; 95% CI: 1.86, 3.80) after adjusting for covariates. The risk of all-cause mortality was significantly greater in participants who experienced weight loss over 4 y (HR: 2.21; 95% CI: 1.32, 3.71) and/or reduced hand grip strength (HR: 1.53; 95% CI: 10.07, 2.17) than in those with stable weight and grip strength, with the highest risk in those with both weight loss and reduced strength (HR: 3.77; 95% CI: 2.54, 5.60).Conclusions: Sarcopenic obesity did not confer any greater risk than sarcopenia alone. Weight loss combined with sarcopenia presented the greatest risk of mortality.


Rapid Profiling of Bovine and Human Milk Gangliosides by Matrix-Assisted Laser Desorption/Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry.
Gangliosides are anionic glycosphingolipids widely distributed in vertebrate tissues and fluids. Their structural and quantitative expression patterns depend on phylogeny and are distinct down to the species level. In milk, gangliosides are exclusively associated with the milk fat globule membrane. They may participate in diverse biological processes but more specifically to host-pathogen interactions. However, due to the molecular complexities, the analysis needs extensive sample preparation, chromatographic separation, and even chemical reaction, which makes the process very complex and time-consuming. Here, we describe a rapid profiling method for bovine and human milk gangliosides employing matrix-assisted desorption/ionization (MALDI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS). Prior to the analyses of biological samples, milk ganglioside standards GM3 and GD3 fractions were first analyzed in order to validate this method. High mass accuracy and high resolution obtained from MALDI FTICR MS allow for the confident assignment of chain length and degree of unsaturation of the ceramide. For the structural elucidation, tandem mass spectrometry (MS/MS), specifically as collision-induced dissociation (CID) and infrared multiphoton dissociation (IRMPD) were employed. Complex ganglioside mixtures from bovine and human milk were further analyzed with this method. The samples were prepared by two consecutive chloroform/methanol extraction and solid phase extraction. We observed a number of differences between bovine milk and human milk. The common gangliosides in bovine and human milk are NeuAc-NeuAc-Hex-Hex-Cer (GD3) and NeuAc-Hex-Hex-Cer (GM3); whereas, the ion intensities of ganglioside species are different between two milk samples. Kendrick mass defect plot yields grouping of ganglioside peaks according to their structural similarities. Gangliosides were further probed by tandem MS to confirm the compositional and structural assignments. We found that only in human milk gangliosides was the ceramide carbon always even numbered, which is consistent with the notion that differences in the oligosaccharide and the ceramide moieties confer to their physiological distinctions.


Exposure to secondhand aerosol of electronic cigarettes in indoor settings in 12 European countries: data from the TackSHS survey.
Exposure to secondhand aerosol from e-cigarette (SHA) may pose harmful effects to bystanders. This study aims to investigate the prevalence, duration and determinants of SHA exposure in various indoor settings in 12 European countries. In 2017-2018, we conducted a cross-sectional study, the TackSHS survey, on a representative sample of the population aged ≥15 years in 12 European countries (Bulgaria, England, France, Germany, Greece, Ireland, Italy, Latvia, Poland, Portugal, Romania and Spain). We described the prevalence and duration of exposure to SHA in several indoor settings among 11 604 e-cigarette non-users. Individual-level and country-level characteristics associated with SHA exposure were also explored using multilevel logistic regression analyses. Overall, 16.0% of e-cigarette non-users were exposed to SHA in any indoor setting at least weekly, ranging from 4.3% in Spain to 29.6% in England. The median duration of SHA exposure among those who were exposed was 43 min/day. 'Other indoor settings' (eg, bar and restaurant) was reported as the place where most of e-cigarette non-users were exposed (8.3%), followed by workplace/educational venues (6.4%), home (5.8%), public transportation (3.5%) and private transportation (2.7%). SHA exposure was more likely to occur in certain groups of non-users: men, younger age groups, those with higher level of education, e-cigarette past users, current smokers, those perceiving SHA harmless and living in countries with a higher e-cigarette use prevalence. We found inequalities of SHA exposure across and within European countries. Governments should consider extending their tobacco smoke-free legislation to e-cigarettes to protect bystanders, particularly vulnerable populations such as young people. NCT02928536.